Acyl-CoA-binding protein/diazepam-binding inhibitor gene and pseudogenes. A typical housekeeping gene family.

Acyl-CoA-binding protein (ACBP) is a 10 kDa protein isolated from bovine liver by virtue of its ability to bind and induce the synthesis of medium-chain acyl-CoA esters. Surprisingly, it turned out to be identical to a protein named diazepam-binding Inhibitor (DBI) claimed to be an endogenous modulator of the GABAA receptor in brain membranes. ACBP/DBI, or proteolytically derived polypeptides of ACBP/DBI, have also been implicated in the control of steroidogenesis in mitochondria and glucose-stimulated insulin secretion. Thus, it appears that ACBP/DBI is a remarkable, versatile protein. Now we have molecularly cloned and characterized the ACBP/DBI gene family in rat. The rat ACBP/DBI gene family comprises one expressed gene and four processed pseudogenes of which one was shown to exist in two allelic forms. The expressed gene is organized into four exons and three introns. There is a remarkable correspondence between the structural modules of ACBP/DBI as determined by 1H nuclear magnetic resonance spectroscopy and the exon-intron architecture of the ACBP/DBI gene. Detailed analyses of transcription of the ACBP/DBI gene in brain and liver were performed to map transcription initiation sites and to examine if transcripts from the ACBP/DBI gene were subject to alternative processing. In both brain and liver, transcription is initiated from two major and multiple minor initiation sites. No evidence for alternative splicing was obtained. The promoter region of the ACBP/DBI gene is located in a CpG island and lacks a canonical TATA box. Thus, the ACDB/DBI gene exhibits all the hallmarks of a typical housekeeping gene.


Mathematical characterization of Chaos Game Representation. New algorithms for nucleotide sequence analysis.

Chaos Game Representation (CGR) can recognize patterns in the nucleotide sequences, obtained from databases, of a class of genes using the techniques of fractal structures and by considering DNA sequences as strings composed of four units, G, A, T and C. Such recognition of patterns relies only on visual identification and no mathematical characterization of CGR is known. The present report describes two algorithms that can predict the presence or absence of a stretch of nucleotides in any gene family. The first algorithm can be used to generate DNA sequences represented by any point in the CGR. The second algorithm can simulate known CGR patterns for different gene families by setting the probabilities of occurrence of different di- or trinucleotides by a trial and error process using some guidelines and approximate rules-of-thumb. The validity of the second algorithm has been tested by simulating sequences that can mimic the CGRs of vertebrate non-oncogenes, proto-oncogenes and oncogenes. These algorithms can provide a mathematical basis of the CGR patterns obtained using nucleotide sequences from databases.


Construction of a microphage variant of filamentous bacteriophage.

The intergenic region in the genome of the Ff class of filamentous phage (comprising strains fl, fd and M13) genome constitutes 8% of the viral genome, and has essential functions in DNA replication and phage morphogenesis. The functional domains of this region may be inserted into separate sites of a plasmid to function independently. Here, we demonstrate the construction of a plasmid containing, sequentially, the origin of (+)-strand synthesis, the packaging signal and a terminator of (+)-strand synthesis. When host cells harboring this plasmid (pLS7) are infected with helper phage they produce a microphage particle containing all the structural elements of the mature, native phage. The microphage is 65 A in diameter and about 500 A long. It contains a 221-base single-stranded circle of DNA coated by about 95 copies of the major coat protein (gene 8 protein).


Pyrrolidine ring puckering in cis and trans-proline residues in proteins and polypeptides. Different puckers are favoured in certain situations.

In a set of proteins studied at high resolution by X-ray crystallography over a half of all cis and trans-proline residues could be unambiguously assigned to one of the two forms of pyrrolidine ring puckering, called UP and DOWN. Of these, 89% of the cis-proline residues exhibit the DOWN pucker, while the trans-proline residues, on average, are about evenly distributed between the two forms. Of trans-proline residues found in alpha-helices, 79% have the UP ring pucker. trans-proline residues occurring in other situations are more equally distributed between the two forms of pucker, although further generalizations may be possible. Proline residues in a set of crystal structures of short polypeptides were also examined. As in the protein sample, a tendency for the cis-proline residues to have the DOWN pucker was observed, but the effect was less pronounced.


Analysis of the role of phosphate oxygens in the group I intron from Tetrahymena.

We have developed a quantitative substitution interference technique to examine the role of Pro-Rp oxygens in the phosphodiester backbone of RNA, using phosphorothioates as a structural probe. This approach is generally applicable to any reaction involving RNA in which the precursor and reaction products can be separated. We have applied the technique to identity structural requirements in the group I intron from Tetrahymena thermophila for catalysis of hydrolysis at the 3' splice site; 44 phosphate oxygens are important in 3' splice site hydrolysis. These include four or five oxygens previously observed to be important in exon ligation. Although phosphate oxygens having a functional significance can be found throughout the intron, the strongest phosphorothioate effects are closely associated with positions in the highly conserved intron core, which are likely to be involved in tertiary interactions, substrate recognition and catalysis.


Differential binding of wild-type and a mutant RepA protein to oriR sequence suggests a model for the initiation of plasmid R1 replication.

DNA replication of the enterobacterial plasmid R1 is initiated by RepA protein. We have developed a new procedure for the purification of RepA from inclusion bodies, which involves CHAPS-mediated solubilization. This method has been also used for the thermosensitive mutant protein RepA2623. The nucleoprotein complexes obtained with both proteins and oriR, the origin of replication, are studied in this paper. DNaseI and hydroxyl-radical footprinting suggest the presence in oriR of two sites with different affinity for RepA separated by eight helical turns. The pattern of hypersensitive sites in the footprints indicates that the oriR sequence, when complexed with RepA, is curved. The binding of RepA molecules to oriR is co-operative and this co-operativity is defective in the thermosensitive protein. Band-shift analysis of RepA-oriR complexes revealed the existence of a species with an anomalously high electrophoretic mobility that appears after formation of the first RepA-oriR complex and requires the sequential interaction of RepA with its two distal binding sites. These features lead us to propose that protein-protein interactions between RepA bound to both distal sites could be responsible for oriR looping. This model represents a novel mechanism that results in activation of an origin in a replicon that does not contain iterons.


In vitro evolution of intrinsically bent DNA.

DNA fragments which are intrinsically bent or curved migrate anomalously during electrophoresis through polyacrylamide gels. Starting with an initial population of approximately 10(12) unique DNA sequences, DNA which exhibited the kind of anomalous mobility associated with DNA bending was selected and enriched using a variation of the SELEX procedure. After seven rounds of selection and amplification, the vast majority of the remaining population of DNA fragments migrated as bent DNA. Cloning and sequencing of 30 individual sequences from this population has yielded information regarding the relationship between DNA sequence and bending. Some of the previous conclusions on DNA bending have been confirmed while others have been modified, by the results presented here. In addition, the dinucleotide base step CA/TG, which had not been thought to be a major factor in DNA bending, appears to be important.


Mutational spectra in human B-cells. Spontaneous, oxygen and hydrogen peroxide-induced mutations at the hprt gene.

To test the hypothesis that reactive species in the oxygen cascade are responsible for spontaneous mutation, we examined the spectra of oxygen and hydrogen peroxide-induced mutations at the hprt locus in a human B-lymphoblastoid cell line. We compared these spectra with the spontaneous mutational spectrum. Large gene alterations were studied by Southern analysis of individual TGR clones. A combination of high fidelity polymerase chain reaction, denaturing gradient gel electrophoresis and direct DNA sequencing were used to detect and identify point mutations in exon 3 of hprt. With regard to spontaneous mutations, a previous study showed that 39% of the spontaneous TGR clones had large gene alterations. In the present study, the analysis of spontaneous point mutations within exon 3 revealed two hotspots. A one base-pair deletion (-A) at base-pair 256 or 257 and a two base-pair deletion (-GG) at base-pair 237 and 238, were detected in triplicate cultures. Each of the hotspots comprised about 1% of the TGR mutants. The analysis of individual oxygen-induced TGR clones (48 h, 910 microM-O2) showed 43% had large gene alterations similar to the spontaneous TGR clones. However, none of the spontaneous point mutation hotspots was found among triplicate oxygen-treated cultures. Two point mutations in common with H2O2-treated cultures were found in one of the three oxygen-treated cultures. Hydrogen peroxide-induced mutations (1 h, 20 microM) also differed from spontaneous mutations. Only 24% of the hydrogen peroxide-induced TGR clones had large gene alterations. The analysis of point mutations showed three hotspots within exon 3 of hprt. An AT to TA transversion at base-pair 259 had an average frequency of 3% of all TGR mutants (present in all of 3 H2O2-treated cultures). Two GC to CG transversions at base-pairs 243 and 202 were present at a frequency of 0.6% and 0.4%, respectively. A five base-pair deletion (base-pair 274 to 278) was present at an average frequency of 0.3%. The latter three mutations were detected in two of three H2O2-treated cultures. Thus, the point mutation spectra of both oxygen and hydrogen peroxide were significantly different from the spontaneous spectrum. The oxygen and hydrogen peroxide-induced spectra shared some features, suggesting that oxygen and hydrogen peroxide share some but not all pathways for induction of mutations within the DNA sequence studied here.(ABSTRACT TRUNCATED AT 400 WORDS)


Structure analysis of the 5' external transcribed spacer of the precursor ribosomal RNA from Saccharomyces cerevisiae.

Full-length precursor ribosomal RNA molecules were produced in vitro using as a template, a plasmid containing the yeast 35 S pre-rRNA gene under the control of the phage T3 promoter. The higher-order structure of the 5'-external transcribed spacer (5' ETS) sequence in the 35S pre-rRNA molecule was studied using dimethylsulfate, 1-cyclohexyl-3-(2-morpholinoethyl)-carbodiimide metho-p-toluenesulfonate, RNase T1 and RNase V1 as structure-sensitive probes. Modified residues were detected by primer extension. Data produced were used to evaluate several theoretical structure models predicted by minimum free-energy calculations. A model for the entire 5'ETS region is proposed that accommodates 82% of the residues experimentally shown to be in either base-paired or single-stranded structure in the correct configuration. The model contains a high degree of secondary structure with ten stable hairpins of varying lengths and stabilities. The hairpins are composed of the Watson-Crick A.T and G.C pairs plus the non-canonical G.U pairs. Based on a comparative analysis of the 5' ETS sequence from Saccharomyces cerevisiae and Schizosaccharomyces pombe, most of the base-paired regions in the proposed model appear to be phylogenetically supported. The two sites previously shown to be crosslinked to U3 snRNA as well as the previously proposed recognition site for processing and one of the early processing site (based on sequence homology to the vertebrate ETS cleavage site) are located in single-stranded regions in the model. The present folding model for the 5' ETS in the 35 S pre-rRNA molecule should be useful in the investigations of the structure, function and processing of pre-rRNA.


Fate of the SpoIIID switch protein during Bacillus subtilis sporulation depends on the mother-cell sigma factor, sigma K.

Sporulation of Bacillus subtilis involves the differentiation of two cell types, the mother cell and the forespore. Two key regulators of mother-cell gene expression are SpoIIID, a DNA-binding protein that activates or represses transcription of many different genes, and sigma K, a subunit of RNA polymerase that directs the enzyme to transcribe genes encoding proteins that form the spore coat. Previous studies showed that SpoIIID is needed to produce sigma K, but suggested that SpoIIID represses sigma K-directed transcription of genes encoding spore coat proteins. Here we show that a feedback loop connects the levels of sigma K and SpoIIID, such that production of sigma K leads to a decrease in the level of SpoIIID. The existence of the feedback loop was demonstrated by using antibodies prepared against SpoIIID to measure the level of SpoIIID during sporulation of wild-type cells, mutants defective in sigma K production, and a mutant engineered to produce sigma K earlier than normal. The feedback loop operates at the level of synthesis and/or stability of spoIIID mRNA, as demonstrated by measuring the level of spoIIID mRNA during sporulation of wild-type cells and mutants defective in sigma K production. Our results suggest that a rise in the level of sigma K during the stage (IV) of spore cortex formation causes a decrease in the level of SpoIIID, which, at least in part, establishes the switch to the stage V (spore coat formation) pattern of mother-cell gene expression.


Tetrahymena gene encodes a protein that is homologous with the liver-specific F-antigen and associated with membranes of the Golgi apparatus and transport vesicles.

The F-antigen is a prominent liver protein which has been extensively used in studies on natural and induced immunological tolerance. However, its intracellular localization and biological function have remained elusive. It has generally been assumed that the F-antigen is confined phylogenetically to vertebrates. Now we have cloned and characterized a gene from the ciliated protozoan Tetrahymena thermophila encoding a protein which clearly is homologous with the rat F-antigen. The coding region of the Tetrahymena F-antigen (TF-ag) gene specifies a 46,051 M(r) protein and is interrupted by three introns. In accordance with the predicted molecular mass of the TF-ag protein, antibodies raised against a cro-lacZ'-TF-ag fusion protein specifically recognized a 45,000 M(r) protein in Western blots of total T. thermophila protein. Immunoelectron microscopy demonstrated that the TF-ag is associated with membranes of the Golgi apparatus and transport vesicles pointing to a role of TF-ag in membrane trafficking. Transcription of the TF-ag gene, as determined by run-on analyses, was only detectable in growing cells, and following transfer to starvation condition pre-existing TF-ag mRNA was rapidly degraded. The abundance of the TF-ag protein, however, declined only moderately during prolonged periods of starvation demonstrating that extensive release of the TF-ag did not take place. In combination these results suggest that the TF-ag protein is a recycled constituent of the intracellular membrane network in T. thermophila.


Selection of high affinity RNA ligands to the bacteriophage R17 coat protein.

RNA ligands with high affinity for the bacteriophage R17 coat protein were isolated from a pool of random RNA molecules using SELEX. Of the 38 ligands isolated, 36 were found to contain a hairpin very similar to the naturally occurring coat protein binding site in the R17 genome. The common features of these 36 sequences provide a consensus binding site and predict components of a hairpin that promote favorable interaction with the coat protein. These include a tetraloop of primary sequence AUCA and a variable-length stem with a bulged adenosine residue at a specific stem position. The predicted consensus agrees well with the highest-affinity RNA binding site of the R17 coat protein, identified through classical but laborious techniques. These results demonstrate the value of SELEX as a tool for isolating high affinity RNA ligands to a specific target protein, and the further value of those ligands to point the researcher toward natural sequences for that target protein.


Conformational changes of a viral capsid protein. Thermodynamic rationale for proteolytic regulation of bacteriophage T4 capsid expansion, co-operativity, and super-stabilization by soc binding.

We have used differential scanning calorimetry in conjunction with cryo-electron microscopy to investigate the conformational transitions undergone by the maturing capsid of phage T4. Its precursor shell is composed primarily of gp23 (521 residues): cleavage of gp23 to gp23* (residues 66 to 521) facilitates a concerted conformational change in which the particle expands substantially, and is greatly stabilized. We have now characterized the intermediate states of capsid maturation; namely, the cleaved/unexpanded, state, which denatures at tm = 60 degrees C, and the uncleaved/expanded state, for which tm = 70 degrees C. When compared with the precursor uncleaved/unexpanded state (tm = 65 degrees C), and the mature cleaved/expanded state (tm = 83 degrees C, if complete cleavage precedes expansion), it follows that expansion of the cleaved precursor (delta tm approximately +23 degrees C) is the major stabilizing event in capsid maturation. These observations also suggest an advantage conferred by capsid protein cleavage (some other phage capsids expand without cleavage): if the gp23-delta domains (residues 1 to 65) are not removed by proteolysis, they impede formation of the stablest possible bonding arrangement when expansion occurs, most likely by becoming trapped at the interface between neighboring subunits or capsomers. Icosahedral capsids denature at essentially the same temperatures as tubular polymorphic variants (polyheads) for the same state of the surface lattice. However, the thermal transitions of capsids are considerably sharper, i.e. more co-operative, than those of polyheads, which we attribute to capsids being closed, not open-ended. In both cases, binding of the accessory protein soc around the threefold sites on the outer surface of the expanded surface lattice results in a substantial further stabilization (delta tm = +5 degrees C). The interfaces between capsomers appear to be relatively weak points that are reinforced by clamp-like binding of soc. These results imply that the "triplex" proteins of other viruses (their structural counterparts of soc) are likely also to be involved in capsid stabilization. Cryo-electron microscopy was used to make conclusive interpretations of endotherms in terms of denaturation events. These data also revealed that the cleaved/unexpanded capsid has an angular polyhedral morphology and has a pronounced relief on its outer surface. Moreover, it is 14% smaller in linear dimensions than the cleaved/expanded capsid, and its shell is commensurately thicker.


Terminating a macromolecular helix. Structural model for the minor proteins of bacteriophage M13.

Analysis of the results of X-ray diffraction, electron microscopy and s sequence studies of filamentous bacteriophage M13 are used to construct structural models for the minor proteins gp7 and gp9 at the end of the virus assembled first, and a portion of gp6 at the end of the virus that binds host. Comparison of the sequence of the major coat protein, gp8, with those of gp7, gp9 and gp6 indicates that significant portions of these three proteins have sequences similar to that of gp8. Assuming that sequence similarity is indicative of structural similarity, gp7, gp9 and portions of gp6 are modeled based on what is known about the structure of gp8. These molecular models are analyzed to predict the packing of the minor proteins with the terminal gp8 proteins (the last gp8 proteins at either end of the helix). This analysis indicates that the gp8 proteins integrated into the virus first may have a structure distinct from those in the body of the virus particle. The gp8 proteins at the end assembled last appear to have a conformation very similar to that of the integral coat proteins. These models place specific constraints on models for the process of viral assembly.


Crystal structure of a sweet tasting protein thaumatin I, at 1.65 A resolution.

The crystal structure of thaumatin I, a potently sweet protein isolated from the fruits of the West African shrub, Thaumatococcus danielli Benth, has been refined at a resolution better than 1.65 A using a combination of energy minimization and stereochemically restrained least-squares methods. The final model consists of all 207 amino acids, 28 alternate amino acid conformers and 236 waters, with a crystallographic R-factor of 0.145 for 19,877 reflections having F > 4 sigma F between 10.0 A and 1.65 A (R = 0.167 for all 24,022 reflections). The model has good stereochemistry, with root-mean-square deviations from ideal values for bond and angle distances of 0.014 A and 0.029 A, respectively. The estimated root-mean-square co-ordinate error is 0.15 A. The current model confirms the previously reported 3.1 A C alpha trace in both main chain connectivity and disulfide topology, including two disulfide bonds, that differed from the earlier reported biochemical determination. The structure contains three domains. The core of the molecule consists of an eleven-stranded, flattened beta-sandwich folded into two Greek key motifs. All beta-strands in this sandwich are antiparallel except the parallel N-terminal and the C-terminal strands. The average hydrogen bond length in this sandwich is 2.89 A, with an angle of 155.1 degrees. Two beta-bulges are found in one of the sheets. The second domain consists of two beta-strands forming a beta-ribbon and connected by an omega-loop, and contains a proline residue in cis conformation. This structural motif folds back against the main sandwich to form a smaller sandwich-like structure. The third domain is a disulfide-rich region stretching away from the sandwich portion of the molecule. It contains one alpha-helix and three short helical fragments. Two of the helical segments are connected by an unusually sharp turn, stabilized by a disulfide bridge. One of the three disulfide bonds in this domain takes on two conformations.


Taming cut-off induced artifacts in molecular dynamics studies of solvated polypeptides. The reaction field method.

In this paper we present a model system of a solvated polypeptide, which is a suitable reference platform for the systematic exploration of methods for taming artifacts introduced by an incorrect treatment of long-range Coulomb forces. The essential feature of the system composed of an alpha-helical peptide and 1021 water molecules is the strict neutrality of all charge groups. The dynamical properties of the peptide, i.e. unfolding or maintenance of the helix, already give first hints on the influence of boundary effects. A rigorous and deeper insight is gained, however, if analyzing the system by means of the generalized Kirkwood g-factor, which projects the net dipole moment of concentric spheres onto the respective dipole moment of the reference charge group. The g-factor is a global measure for, and a sensitive probe of, the orientational structure, which in its turn reflects even the smallest inconsistencies in the treatment of long-range forces. While the cut-off scheme failed the g-factor test, the "reaction field" method, the simplest cut-off correction scheme, enables a consistent description. In other words, with the aid of the reaction field, the correct orientational structure is restored. As a consequence, the helix stability is regained and we were able to calculate the dielectric constant epsilon approximately 55 to 60 for our system, which is slightly below the corresponding value epsilon SPC = 66 of the pure solvent.


Analysis of sequence variation among legume lectins. A ring of hypervariable residues forms the perimeter of the carbohydrate-binding site.

Twelve plant lectins from the Papilionoideae subfamily were selected to represent a range of carbohydrate specificities, and their sequences were aligned. Two variability indices were applied to the aligned sequences and the results were analysed using the three-dimensional structures of concanavalin A and the pea lectin. The areas of greatest variability were located in the carbohydrate-binding site region, forming a perimeter around a well-conserved core. These residues are inferred to be specificity determining, in the manner of antibodies, and the most variable position corresponded to Tyr100 in concanavalin A, a known ligand contact residue. In addition to the five peptide loops known to form the binding site from crystallographic studies, a sixth segment with variable residues was located in the binding-site region, and this may contribute to oligosaccharide specificity. In their overall composition, the lectin sites resemble those of the sugar-transport proteins rather than antibodies. The prospects for modelling lectin binding sites by the methods used for antibodies were also assessed.


Isolation, crystallization, crystal structure analysis and refinement of B-phycoerythrin from the red alga Porphyridium sordidum at 2.2 A resolution.

The light-harvesting pigment-protein complex B-phycoerythrin from the red alga Porphyridium sordidum has been isolated and crystallized. B-Phycoerythrin consists of three different subunits forming an (alpha beta)6 gamma aggregate. The three-dimensional structure of the (alpha beta)6 hexamer was solved by Patterson search techniques using the molecular model of C-phycocyanin from Fremyella diplosiphon. The asymmetric unit of the crystal cell (space group P3, with a = b = 111.2 A, c = 59.9 A, alpha = beta = 90 degrees, gamma = 120 degrees) contains two (alpha beta) monomers related by a local dyad. Three asymmetric units are arranged around the crystallographic 3-fold axis building an (alpha beta)6 hexamer, as in C-phycocyanin. The crystal structure has been refined by energy-restrained crystallographic refinement and model building. The conventional R-factor of the final model was 18.9% with data to 2.2 A resolution. The molecular structures of the alpha and beta-subunits resemble those of C-phycocyanin. Major changes in comparison to phycocyanin are caused by deletion or insertion of segments involved in protein-chromophore interactions. The singly linked phycoerythrobilin chromophores alpha-84, alpha-140a, beta-84 and beta-155 are each covalently bound to a cysteine by ring A. The doubly linked chromophore beta-50/beta-61 is attached at cysteine beta-50 through ring A and at cysteine beta-61 through ring D. B-Phycoerythrin contains additionally a 30 kDa gamma-subunit, which is presumably located in the central cavity of the hexamer. It is disordered, as a consequence of crystal and local symmetry averaging.


Recognition of distantly related protein sequences using conserved motifs and neural networks.

A sensitive technique for protein sequence motif recognition based on neural networks has been developed. It involves three major steps. (1) At each appropriate alignment position of a set of N matched sequences, a set of N aligned oligopeptides is specified with preselected window length. N neural nets are subsequently and successively trained on N-1 amino acid spans after eliminating each ith oligopeptide. A test for recognition of each of the ith spans is performed. The average neural net recognition over N such trials is used as a measure of conservation for the particular windowed region of the multiple alignment. This process is repeated for all possible spans of given length in the multiple alignment. (2) The M most conserved regions are regarded as motifs and the oligopeptides within each are used to train intensively M individual neural networks. (3) The M networks are then applied in a search for related primary structures in a databank of known protein sequences. The oligopeptide spans in the database sequence with strongest neural net output for each of the M networks are saved and then scored according to the output signals and the proper combination that follows the expected N- to C-terminal sequence order. The motifs from the database with highest similarity scores can then be used to retrain the M neural nets, which can be subsequently utilized for further searches in the databank, thus providing even greater sensitivity to recognize distant familial proteins. This technique was successfully applied to the integrase, DNA-polymerase and immunoglobulin families.


Unique side-chain conformation encoding for chirality and azimuthal orientation in the molecular packing of skin collagen.

We used molecular mechanics to study the role of gly X-Y+ sequences, where X- was Asp or Glu and Y+ was Lys or Arg, in the molecular packing of type I collagen. In the minimal energy conformation of a triply stranded molecule having a coiled-coil configuration, the side-chains of these sequences segregated into two oppositely charged groupings of the forms X-Y+X- and Y+X-Y+. Groupings having the same net charge were clustered along two complementary azimuthal edges of the molecule. Intermolecular interactions, through these oppositely charged edges, align the molecules appropriately for the formation of the HHL crosslink of skin. This alignment also can account for the axial periodicity and chiral appearance of skin collagen fibrils.


Crystallization and preliminary X-ray analysis of a fungal endoglucanase I.

An endoglucanase I (EG1) from a fungal source (Humicola insolens) has been crystallized in a number of forms suitable for X-ray diffraction analysis. Four crystal forms have been grown from various precipitants using vapour phase diffusion methods in hanging drops. Three of these crystal forms diffract to beyond 2.5 A resolution. Two forms, obtained from ammonium sulphate at pH 5.4, or 8.0, grow as tetragonal bipyramids in space group P4(1)22 or P4(3)22, with approximate cell dimensions a = b = 102 A, c = 282 A. The other crystal forms were grown from polyethylene glycol 8000 at pH 8.0. One grows as monoclinic plates, space group P2(1), with cell dimensions a = 66.9 A, b = 75.2 A, c = 86.9 A and beta = 102.9 degrees and the other as long hexagonal rods in space group P6(1)22 or P6(5)22, with cell dimensions a = b = 119 A, c = 83 A.


Preliminary crystallographic studies of the amino terminal half of human lactoferrin in its iron-saturated and iron-free forms.

The amino terminal half of human lactoferrin (LfN) produced from transfected baby hamster kidney cells has been crystallized in its iron-saturated and iron-free forms. The crystals of glycosylated LfN and deglycosylated LfN are monoclinic, space group C2, with cell dimensions a = 133.0 A, b = 58.3 A, c = 58.3 A, alpha = 90.0 degrees, beta = 114.7 degrees, gamma = 90.0 degrees, and one molecule per asymmetric unit. Crystals of apo LfN have also been prepared using deglycosylated protein. These crystals are tetragonal, space group P4(1)2(1)2 (or P4(3)2(1)2), with cell dimensions of a = b = 58.4 A and c = 217.2 A and one molecule per asymmetric unit. Both the iron-saturated and the iron-free crystals are suitable for high resolution X-ray analysis.


Crystallization of murine major histocompatibility complex class I H-2Kb with single peptides.

X-ray quality crystals of a soluble murine class I H-2Kb molecule complexed with three different peptide antigens were grown in several forms by streak seeding and macroseeding methods. Co-crystals with VSV-8 (RGYVYGQL), OVA-8 (SIINFEKL) and SEV-9 (FAPGNYPAL) peptides were grown either from NaH2PO4/HPO4 or from polyethylene glycol 4000 within the pH range 5.0 to 7.5, with the use of 4-methyl-2-pentane diol (MPD) as an additive. The VSV-8 crystals grew in space groups P1, with cell dimensions a = 63.1 A, b = 69.1 A, c = 72.0 A, alpha = 89.9 degrees, beta = 77.1 degrees, gamma = 123.3 degrees and P2(1)2(1)2, with a = 138.1 A, b = 88.6 A, c = 45.7 A, and diffract to 2.9 and 2.3 A, respectively. Crystals of the SEV-9 complex grew from similar crystallization conditions to those of the orthorhombic VSV-8 complex with similar cell parameters and diffract to at least 2.5 A resolution. Crystals of the OVA-8 complex were obtained from either phosphate (space group C2, a = 118.7 A, b = 61.6 A, c = 85.3 A, beta = 108.4 degrees) or polyethylene glycol (space group P1, a = 64.5 A, b = 71.0 A, c = 66.3 A, alpha = 89.7 degrees, beta = 95.7 degrees, gamma = 123.3 degrees) and diffract to 3 A resolution. The crystallization procedures used here significantly increased the rate and production of X-ray quality crystals.


Purification, crystallization and preliminary X-ray analysis of toxic shock syndrome toxin-1 from Staphylococcus aureus.

High yields of toxic shock syndrome toxin-1, from Staphylococcus aureus, have been purified (> 99%) using a novel, simple, two-step procedure involving dye ligand chromatography. Crystals suitable for X-ray diffraction work were obtained by vapour diffusion using ammonium sulphate and polyethylene glycol as precipitants. They belong to the orthorhombic space group C222(1), with unit cell dimensions a = 108.6 A, b = 177.6 A and c = 97.5 A, with three molecules per asymmetric unit. The crystals diffract to at least 2.5 A resolution and are suitable for three-dimensional X-ray structural analysis.


Crystallization and preliminary X-ray diffraction studies of pancreatic spasmolytic polypeptide.

Pancreatic spasmolytic polypeptide (PSP) isolated from porcine pancreas has been crystallized by the hanging drop vapour diffusion method. The crystals belong to the space group I222 or I2(1)2(1)2(1) with cell dimensions a = 181.9 A, b = 54.5 A, c = 72.9 A. The crystals diffract to at least 2.5 A resolution and the asymmetric unit contains two molecules (Vm = 3.9 A3/Da) with a solvent content of 68% as determined by density measurements of the crystals. The self-rotation function suggests that the two molecules within the asymmetric unit are related by a 2-fold axis at either 30 degrees or 60 degrees from a in a plane perpendicular to the b axis.


Nucleotide sequence and organization of an H2-uptake gene cluster from Rhizobium leguminosarum bv. viciae containing a rubredoxin-like gene and four additional open reading frames.

The nucleotide sequence of a 3.2 kb region following the hydrogenase structural operon (hupSLCDEF) in the H2-uptake gene cluster from Rhizobium leguminosarum by viciae strain 128C53 has been determined. Five closely linked genes encoding products of 16.3 (HupG), 30.5 (HupH), 8.0 (HupI), 18.4 (HupJ) and 38.7 (HupK) kDa were identified 166 bp downstream from hupF. Transposon insertions into hupG, hupH, hupJ and hupK suppress the H2-oxidizing capability of the wild-type strain. The amino acid sequence deduced from hupI contains two Cys-X-X-Cys motifs, characteristic of rubredoxins, separated by 29 amino acid residues showing strong sequence homology with other bacterial rubredoxins. The amino acid-derived sequence from hupG and hupH showed homology to products from genes hyaE and hyaF of the operon encoding hydrogenase 1 from Escherichia coli, and hupJ and hupK were related to open reading frames identified in Rhodobacter capsulatus and Azotobacter vinelandii hydrogenase gene clusters. An involvement of the hupGHIJK gene cluster in redox reactions related to hydrogenase synthesis or activity is predicted on the basis of the function as electron carrier attributed to rubredoxin.


Overview of the red eye.

Markedly reduced vision and a shallow anterior chamber suggest acute glaucoma. A mucopurulent discharge suggests conjunctivitis. An abnormal cornea may suggest acute glaucoma or keratitis. An abnormal pupil is seen with both acute glaucoma and iritis.


A challenge for the 1990s: patient education.

The goal of health education is assisting patients to adapt to their illness, assuming responsibility for their therapeutic regimen, and working towards attaining their maximum level of wellness. There are three domains of learning behavior: cognitive (knowledge and information), affective (attitudes, values, and beliefs), and psychomotor (skills and performance). The use of these results in behavioral change. Effective nurse educators take advantage of every interaction to teach their patients. It is the nurse's responsibility to present information to the patient in such a way that it motivates, stimulates, and allows the patient to appreciate learning.


Corneal edema and bullous keratopathy.

The normally functioning cornea is in a relatively dehydrated or deturgesced state, which maintains the compact anatomic relations of the stroma and epithelium. The corneal endothelium functions as both a barrier and a pump to maintain corneal deturgescence. Even minimal epithelial edema can cause a significant decrease in visual acuity.


OSHA regulations: how they relate to ophthalmic practice.

The OSHA regulations, which took effect March 6, 1992, require that all employees be trained in infection control practices when they are hired, their job description changes, or the standards for universal precautions are revised. An explanation, provided to inform employees where a copy of the OSHA standard can be reviewed should be available at each clinical and surgical site, and OSHA regulation definitions must also be posted. The OSHA regulation applies to any clinical, housekeeping, or administrative staff that has any potential risk of exposure to blood or other potentially infectious substances.


Targeting of gonadotropin-releasing hormone axons from preoptic area grafts to the median eminence.

Implantation of normal GnRH neurons can reverse many of the reproductive deficiencies that characterize hypogonadal (hpg) mice. Since the GnRH axons follow a stereotyped trajectory to their target we investigated the possibility that host brain regions adjacent to the graft might provide signals that induced this directional growth. The role of the adenohypophysis in GnRH axonal outgrowth was studied in mice with co-grafts of fetal preoptic area (POA) and pituitary and in hypophysectomized hosts. When fetal pituitaries were grafted together with the POA, immunoreactive GnRH fibers did enter the glandular tissue but they also grew into the host median eminence. Surgical removal of the pituitary of hpg hosts prior to POA graft placement was also compatible with GnRH innervation of the host median eminence although in some individuals that innervation pattern was confined to the more caudal aspects. The results of these two experiments suggest that the anterior pituitary gland may be an attractive target for GnRH axons but that this tissue is not essential for directed GnRH axonal outgrowth to its target. To determine if the median eminence itself could direct the growth of GnRH axons, co-grafts of POA and a fetal medial basal hypothalamic (MBH) block, which was predominantly median eminence, were made. Immunocytochemistry showed that an intragraft mini-median eminence was formed with a highly organized and robust GnRH innervation. Ultrastructural analysis indicated that these axons terminated near fenestrated capillaries. However, even under these conditions some GnRH axons exited into the host median eminence. It now seems likely that a cellular component of the median eminence can provide a signal to attract GnRH axons. Whether this signal is produced by the specialized ependymal cells, by the endothelia, or by meningeal (pial) components must now be tested.


Central serous chorioretinopathy associated with pregnancy.

Central serous chorioretinopathy is an infrequent but benign complication of pregnancy that is found largely in the minority population. White subretinal exudates and multiple recurrences are frequently seen with the central serous chorioretinopathy of pregnancy. Central serous chorioretinopathy usually spontaneously resolves with minimal sequelae without intervention after delivery of the baby.


Red-green duochrome test.

The red-green duochrome test is based on a monocular endpoint in which each eye is tested separately. It is a subjective test that requires responses from the patient and is used to refine the spherical endpoint. Chromatic aberration, the basis of the test, occurs because different wavelengths of light are bent to a different extent. The longer wavelength (red) is refracted less than the shorter (green). If the letters on the red side stand out more, add minus power; if the letters on the green side stand out more, add plus power. Neutrality is reached when the letters on both backgrounds appear equally distinct.


Neuromuscular junctions contain NP185: the multifunctional protein is located at the presynaptic site.

The NP185 polypeptide (AP3) is a multifunctional component isolated from brain endocytic vesicles, which binds to tubulin and clathrin light chains, decoated vesicles, synaptic vesicles, and the synaptosomal plasma membrane (Su et al., 1991). The NP185 molecules are expressed during avian cerebellar synaptogenesis and appear to function in CNS regions rich in synaptic terminals (Perry et al., 1991). In this report we describe double-labelling experiments with avian embryonic striated muscle fibers demonstrating the exclusive presence of the brain-specific protein at the neuromuscular junction. We used indirect rhodamine immunofluorescence labeling with a monoclonal antibody (mAb-8G8) to mark the location of NP185 in muscle combined with fluorescein-alpha-bungarotoxin to mark the postsynaptic location of the acetylcholine receptors (AChRs). We show that the distribution of both NP185 and AChRs has an overall correlation, but the location of NP185 is circumscribed to presynaptic structures adjacent but not overlapping with postsynaptic structures displaying the AchRs. To confirm the identity of NP185, the molecule was extracted from both tissues, partially purified, immunoprecipitated, and identified in Western blots with the mAb 8G8. The mAb reacted with an identical 185 kD protein band purified from both tissues. Based on its properties and specific neuronal location, the NP185 molecule may function in motor nerve terminals by screening membrane proteins, identifying areas of the synaptic plasma membrane, and to anchor these elements with structural proteins for their recycling and transport within the neuronal cellular compartments.


Amygdala neurons in vitro: neurite growth and effects of estradiol.

Dissociated cell cultures from embryonic rat medial amygdala were studied using sequential photography and immunocytochemical staining for cytoskeletal proteins and substance P (sP). Cultures were seeded with cells taken from fetuses grouped by sex; experimental cultures were raised in medium containing 17-beta-estradiol (E2). Forty-eight hours after plating a few neurons begin to define their morphological polarity by the differentiation of an axon-like process; at 5 days in vitro (DIV) almost all neurons had developed an axon. Tapering, daughter branch ratio and branch power coefficient coincided with identification of dendrites which could be confirmed by retrospective analysis of immunocytochemically stained cultures: at 5 DIV MAP-2 was restricted to dendrites whereas Tau immunoreactivity was differentially localized with a clear predominance in the axon. At 21 DIV neuronal shape parameters were strikingly like those of amygdaloid neurons in vivo. It was demonstrated in living neurons that E2 increased total dendritic length and that this is due to increased ramification of third or higher order dendritic segments whose individual lengths are not different from controls. Densitometric measurement of MAP-2 stained neurons showed a highly significant increase of immunoreactive material in cells grown in the presence of E2; readings for alpha-tubulin were not different between controls and E2 treated cultures. The effect of E2 on dendritic length was just as manifest in sP-positive as in sP-negative neurons. No sexual differences in morphological parameters, growth characteristics or effects of E2 were found in neurons taken from female fetuses versus neurons from male fetuses. The significance of these results for the generation of sexual differences in the amygdala in vivo is discussed and contrasted with reported results on the effects of E2 in cultures of different neural regions.


Beta-oxidation of [1-14C]palmitic acid by mouse astrocytes in primary culture: effects of agents implicated in the encephalopathy of Reye's syndrome.

beta-Oxidation of [1-14C]palmitic acid was examined in homogenates of astrocytes cultured from neonatal mouse brain. Under optimal reaction conditions (< or = 50 micrograms protein, 10 min at 37 degrees C), oxidation increased as a function of palmitate concentration (15 microM to 2 mM) and reached a maximum rate of 1.98 +/- 0.29 nmol/min/mg protein (mean +/- SEM) at 0.2 mM substrate. Eadie-Hofstee analysis of data from four experiments yielded apparent values for Vmax of 1.87 nmol/min/mg protein, and for Km, 35-40 microM. There were no dramatic changes in the oxidation rate in cells between 10 and 36 days in culture. During the 10-min assays, less than 0.05% of the radioactivity was converted to 14CO2 by the astrocytes; water-soluble products accounted for 1-2% of the total substrate added. Studies with KCN indicated that 60-70% of the total activity occurred in the mitochondria. We have been studying the structural and functional changes associated with the cerebral encephalopathy of Reye's syndrome (RS). Three-week-old astrocytes exposed to serum from RS children for the final 7 days of culture exhibited minor mitochondrial pleomorphism and had increased numbers of other intracellular organelles. Examination of the effects of agents implicated in RS indicated that oxidation of [1-14C]palmitate was not altered by Na+ salicylate (1-3 mM), but was inhibited by the industrial surfactant, Toximul MP-8 (> or = 10 micrograms/ml), 4-pentenoic acid (> or = 0.1 microM), or with 4 days' exposure to ammonia (10 nM). The latter treatment also resulted in an increase in protein synthesis, cell volume, and malondialdehyde formation. These results suggest that some of the "toxins" implicated in RS inhibit fatty-acid oxidation in the astrocytes and produce other lipid-related abnormalities that could be related to encephalopathy.


Anomalous phosphorylated neurofilament aggregations in central and peripheral axons of hens treated with tri-ortho-cresyl phosphate (TOCP).

Previous biochemical studies demonstrated a dramatic increase in phosphorylation of cytoskeletal proteins that occurs early in organophosphorus ester-induced delayed neurotoxicity (OPIDN). In this report we present immunohistochemical evidence that there is anomalous aggregation of phosphorylated neurofilaments within central and peripheral axons following organophosphate exposure. The morphology, location, and time of appearance of these aggregations are consistent with the hypothesis that the aberrant phosphorylation of cytoskeletal elements is an antecedent to the focal axonal swelling and degeneration characteristic of OPIDN.


Interaction of ganglioside GM1 with the B subunit of cholera toxin modulates intracellular free calcium in sensory neurons.

The B subunit of cholera toxin, which binds specifically to GM1 ganglioside on cell surfaces, has previously been shown to modulate intracellular calcium levels and growth in several cell types. To explore a role for such changes in calcium in the growth regulatory function of cell-associated GM1 in neurons, dissociated neurons from chicken embryonic day 8 dorsal root ganglia were exposed to the B subunit. To enhance sensitivity to B subunit, some neurons were also enriched with added GM1 (100 microM) and then exposed to B subunit. Incubation of naive cultures with 1 microgram/ml of the B subunit was sufficient to produce modest increases in intracellular free calcium above basal levels in a minor percentage of cells for at least 5 min, as measured by fura-2 fluorescence imaging. Pretreatment of the cells with GM1 for 48 hr increased even further the elevations in intracellular free calcium and the percentage of responding neurons observed after B subunit exposure. These increases in intracellular calcium required the presence of external Ca2+, but were not inhibited by calcium channel blockers. Such changes in calcium were accompanied by fine alterations in morphology affecting mostly the branching of neurites and were more pronounced in the presence of GM1. However, the morphological changes did not result in altered neurofilament protein expression. Immunogold electron microscopy using anti-choleragenoid depicted extensive aggregations of immunoreactive gold particles on neuronal surfaces, which were more extensive in cells treated with GM1. The results demonstrate that cell incorporated GM1 may modulate calcium fluxes, perhaps accounting for the growth regulatory functions of GM1 in both neuronal and other cell types.


Leukemia inhibitory factor promotes the neuronal development of spinal cord precursors from the neural tube.

Recent evidence from our laboratory has shown that leukemia inhibitory factor (LIF) can act early in peripheral nervous system development. We have investigated a potential role of LIF in the developing spinal cord. In explants and dissociated cultures of spinal cord primordium, LIF stimulated a profuse neurite outgrowth. To determine if these effects were related to neuronal differentiation, cells were plated at low cell density and stained for neurofilament. LIF stimulated an increase in the number of newly differentiated neurons, without inducing proliferation of the precursors. Given that LIF has previously reported effects as a cholinergic switching factor for sympathetic neurons, we investigated whether LIF had similar effects in these spinal cord cultures. LIF increased the number of cholinergic neurons in proportion to its overall effect on the stimulation of all neurofilament positive neurons in the culture. These data show that LIF stimulates the generation of spinal cord neurons from their precursors and further implicates a role for LIF in nervous system development.


Development of the ostiomeatal unit in childhood: a radiological study.

The role of surgery in the management of sinus disease is in a state of evolution. With the advent of the endoscope, functional sinus surgery has become a reality. This type of surgery is recently being utilized as a therapeutic modality for sinus disease in children. The sinus surgeon should be aware of the differences between adult and pediatric sinus anatomy. This study utilizes the CT scanner to chart the anatomy and development of the ostiomeatal unit area in childhood, from the age of one through 16. This study may provide a more comprehensive understanding of the ostiomeatal unit in the pediatric population.


Augmentation of skeletal muscle survival in the latissimus dorsi porcine model using acute ischemic preconditioning.

Reconstruction of head and neck defects commonly involve local and distal muscle transplants. Ischemic flap necrosis is a well recognized complication of autogenous muscle transplants. Research in the field of myocardial survival has recently shown that repeated brief periods of ischemia followed by reperfusion result in improve muscle survival when the cardiac muscle is subsequently subjected to prolonged ischemia. This is called preconditioning and the mechanism is unknown. The concept is discussed in this paper and presented as a potential new, non-pharmacological strategy to reduce skeletal muscle ischemic necrosis. A review of the development and application of preconditioning in myocardial muscle survival is presented, and causes of ischemic muscle necrosis are discussed. Potential mechanisms of preconditioning in muscle are also presented. The extrapolation of the concept of preconditioning is discussed with an examination of a potential experimental model to test this concept: the latissimus dorsi flap in the pig. The results of our investigations into the effect of preconditioning on skeletal muscle and its possible mechanism are discussed. Preconditioning may prove to be a useful method to improve skeletal muscle tolerance to sustained normothermic global ischemia. This may have important clinical benefits in muscle transplantation. To our knowledge, this is the first publication to discuss the application of preconditioning in skeletal muscle.


Tissue clay: an experimental model to evaluate its performance in soft tissue augmentation and its biologic fate.

Tissue clay (a mixture of autologous whole blood and microfibrillar collagen--Avitene) has been proposed as a medium for reconstruction in cosmetic and reconstructive deformities of the face. Its performance and biologic fate has not been evaluated experimentally. The current study employs a model using the rabbit pinna to determine, histologically, the fate of a tissue clay implant.


Presbycusis: correlations of clinical audiology with morphological changes in the cochlea and the ventral cochlear nucleus.

Five cases of presbycusis are presented. Analysis included audiological tests, cochlear hair cell and ganglion cell counts, assessment of degree of strial atrophy, and ventral cochlear nucleus neuron counts. One case showed a reduction in cochlear nucleus neuron counts without significant cochlear changes, indicating that this pattern of pathological changes is a possible cause of high tone sensorineural deafness.


Long-term results of uvulopalatopharyngoplasty for snoring.

Snoring is a disease of listeners and one which can cause significant disruption in an otherwise peaceful household. Uvulopalatopharyngoplasty (UPPP) was initially described in 1981 for the treatment of obstructive sleep apnea, and since that time only one large series has evaluated it as a treatment for snoring. Various therapeutic strategies have been described for the treatment of snoring, but UPPP seems to hold the most promise. A retrospective review was undertaken of over 100 patients undergoing UPPP since 1984 to evaluate the efficacy of the procedure in controlling the symptom of snoring. The group of patients who will benefit most from this procedure is identified.


Chondrosarcoma of the larynx: a histo-radiologic analysis.

Chondrosarcoma of the larynx is a rare malignant cartilaginous neoplasm. To date, the serial sectioned laryngeal specimens of this disease have not been correlated with the pre-operative computed tomographic findings. A retrospective analysis of 833 serially sectioned laryngeal specimens found chondrosarcoma in six cases (0.72%), of which four cases had a pre-surgical computed tomography (CT). Radiologic axial sections were compared with microscopic sections in the same plane. Whole organ sections showed the tumor to have an expansile growth pattern, showing little impulse to infiltrate adjacent cartilage. CT scanning successfully predicted the diagnosis of a cartilage neoplasm prior to surgery, and accurately reflected the site and extent of laryngeal involvement. We conclude that CT is a reliable method of evaluating chondrosarcoma of the larynx, thereby predicting the feasibility of partial laryngeal resection.


Bilateral vocal cord palsy: the alternative to tracheostomy.

Bilateral abductor vocal cord paralysis remains a difficult management problem for otolaryngologists. The techniques currently used fall into four main types: arytenoidectomy, cordectomy, cord lateralization and laryngeal re-innervation. These techniques invariably require a tracheostomy either prior to or at the time of the main procedure. A technique for managing patients without performing a tracheostomy which is as effective as the previously mentioned procedures and with no greater morbidity in terms of voice production was initially described by Ejnell, et al (1984). The results in three patients using this technique which involves vocal cord lateralization under endoscopic control confirms its ease and efficacy. All three patients were relieved of their presenting dyspnea and had satisfactory postoperative voice production. This appears to be an effective technique for the treatment of patients with bilateral abductor laryngeal paralysis which obviates the need for tracheostomy and its attendant complications.


Oral leukoplakia caused by cinnamon food allergy.

An oral leukoplakic lesion that was clinically thought to be a squamous cell carcinoma is presented. The cinnamon component of chewing gum was discovered to be the etiologic agent. The case is unusual because of the leukoplakia noted; all other previously reported cases described erythematous patches only. The role of food allergy in the pathophysiology of the lesion is discussed.


Unusual presentation of verrucous carcinoma of maxillary antrum.

Verrucous carcinoma is a rare type of squamous cell carcinoma that is most often seen in the oral cavity and larynx. This paper describes a case report of a rare tumor with an unusual presentation. This patient presented with recurrent pyoceles and was diagnosed by chance. Carcinoma of maxilla is rarely diagnosed until the bony walls are eroded. This condition should be kept in mind in patients presenting with recurrent pyocele and a biopsy carried out from the maxilla.


Experience of haemodialysis at the Kidney Centre.

Seventy-nine patients of end stage renal disease (ESRD) on maintenance haemodialysis were studied. Most of the cases were in their prime of life. The disease was equally common in both sexes and all ethnic groups. Chronic glomerulonephritis was the commonest cause followed by diabetes mellitus. Hypertension was the commonest associated illness. All patients were screened for hepatitis B surface antigen and antibody and those found negative were vaccinated. A-V fistula in the upper extremity was used as the vascular access in 93% cases. In 68% cases dialyzer was reused without any ill effect. Amongst the complications observed, hypotension was seen in 65%, psychological disorders in 52%, followed by nausea, vomiting, itching and cramps. Technical complications were related to A-V fistula in 45% cases. Forty three percent patients were maintained without blood transfusion and 88% showed improvement in their quality of life.


Non-specific reactive hyperplasia of cervical lymph nodes: a follow-up.

Non-specific reactive hyperplasia (NSRH) accounted for 20% patients of cervical lymphadenopathy collected over a period of 1-1/2 years. The disease affected comparatively younger people of both sexes equally. Routine investigations offered little diagnostic help. The disease resolved spontaneously in 1/3rd of patients, 1/4th developed specific entities, while in the rest it continued as a prolonged ailment. Lymph node biopsy should be planned meticulously to avoid missed diagnosis. If the lymphadenopathy of NSRH does not subside spontaneously within two months, a re-biopsy is advised to pick up a specific ailment.


Byssinosis in ginning factories of rural Sindh.

Frequency of byssinosis in 276 workers from 5 ginning factories of rural Sindh are described. Twenty-four (9%) had byssinosis, 178 were asymptomatic and the remaining 74 had other respiratory symptoms. Chronic obstructive pulmonary disease was present both in symptomatic and asymptomatic workers and was equally distributed amongst smokers and non-smokers. A possible involvement of some factors other than cigarette smoking is discussed.


[The signal transduction mechanism responsible for interferon-gamma-inducible indoleamine 2,3-dioxygenase (IDO) gene expression in T98G cells].

The interferon (IFN)-gamma-induced indoleamine 2,3-dioxygenase (IDO) is implicated in the inhibition of intracellular pathogens, e.g. Chlamydia psittaci and Toxoplasma gondii. The intracellular signaling molecules responsible for the induction of IDO gene expression were investigated by the quantitative polymerase chain reaction. The gene expression was inhibited by a tyrosine kinase inhibitor, genistein. Being consistent with this, IFN-gamma induced increased tyrosine phosphorylation and this was inhibited by genistein. The transcription of IDO gene was not inhibited by protein kinase C (PKC) inhibitors, H-7 and staurosporine, or a calmodulin inhibitor, W-7. Irrelevance of PKC in IDO gene expression was supported by the failure of PMA or PMA + A23187 to induce IDO gene expression. These results all suggest that the tyrosine phosphorylation is a critical event in IFN-gamma-inducible IDO gene expression and PKC is not involved in the gene expression.


[Prevalence and pathogenesis of iron deficiency in Japanese women (1981-1991)].

Iron deficiency anemia is the most prevalent among form of anemia in the world. In Japan, there is no overall report concerning prevalence and pathogenesis of iron deficiency. We estimated the prevalence of iron deficiency from the results of a survey of 3,015 Japanese women. The reference range for hemoglobin was derived from the average value of subjects with normal iron status (> or = 16% of transferrin saturation and > or = 12 ng/ml of serum ferritin). Using these reference standards, the prevalence of iron deficiency anemia, latent iron deficiency, storage iron deficiency, normal and others were 8.5%, 8.0%, 33.4%, 43.6% and 6.5%, respectively. The prevalence of iron deficiency anemia increased beginning in early lower teen girls, was highest in high teen-girls and, young women and decreased in elderly women. In elderly women, the cause of iron deficiency was often obvious associated with anemia of chronic disorders. The strategy for iron deficiency was discussed concerning iron fortification, mega-ingestion of vitamin C and low dose administration of iron tablets in the higher incidence group.


[Evaluation of hypercoagulable state in patients with malignancies by using prothrombin fragment F1 + 2].

In order to elucidate the activation of the coagulation cascade in patients with malignant neoplasms, we measured the levels of plasma prothrombin fragment F1 + 2, which is liberated in the process of thrombin generation. Twenty healthy adults (Group A), 29 patients with malignancies not complicated with DIC (Group B) and 4 patients with DIC (Group C) were evaluated. The values of F1 + 2 in Group C (2.38 +/- 0.55 nmol/l) were significantly higher (p < 0.01) than those in Group A (0.52 +/- 0.19 nmol/l) and B (0.86 +/- 0.68 nmol/l). Many patients in Group B showed higher levels of F1 + 2 compared to normal subjects, however, no significant differences were found between Group A and B. With respect to other coagulation molecular markers such as TAT, D-Dimer and PIC, F1 + 2 levels revealed positive correlation to those levels. Concerning the clinical course of DIC, elevated levels of F1 + 2 normalized much rapidly than those of TAT and D-Dimer by continuous administration of heparin. In conclusion, the measurement of plasma F1 + 2 is important in monitoring the activation of coagulation system in patients with malignancies, especially with respect to early detection and treatment of DIC.


[Differentiating effect of oral administration of retinol palmitate (Chocola-A) for an aged AML (M3) with severe complications].

A 74 year-old woman, who had been diagnosed as AML (M3) in poor condition, was treated with Retinol Palmitate (Chocola-A, 150,000 unit/m2 per os, after informed consent. An increase of white blood cells (neutrophil) counts was observed after 7 days. After 4 weeks, WBC counts were increased to 20,700/microliters (neutrophil counts 6,400/microliters) Maturation tendency of leukemic cells was also proved in the bone marrow. In vitro studies showed that morphological differentiation was recognizable in cultured leukemic cells treated with 10(-6)M all-trans retinoic acid after 6 days, but not in controls. Responses in the NBT reduction test were slightly less than in the clinical study. The administration of Retinol Palmitate may be a new regimen to treat AML (M3) in aged patients in poor condition.


[Extramedullary diseases as presenting features of aleukemic acute monocytic leukemia].

We present a case of acute monocytic leukemia presenting with extramedullary disease clinically resembling lymphoma. A 36 year-old man presented with arthralgia and was found to have skin eruption, nasopharyngeal mass, hepatosplenomegaly and superficial lymphadenopathy. The biopsies of those lesions as well as bone marrow revealed infiltration of CD45 positive large atypical cells and were interpreted as malignant lymphoma. At that time blood film showed leuko-erythroblastic picture, but no atypical cells were noted. He was treated with CHOP followed by VEPA chemotherapy and achieved partial remission. Four months later headache and double vision occurred with the same atypical cell in the cerebrospinal fluid. The cells were identified as monoblasts by cytochemistry and immunophenotype. The diagnosis of aleukemic acute monocytic leukemia was made and its CNS involvement was successfully treated with chemotherapy with DHAP and intrathecal MTX. However, two months later, blasts finally appeared in the peripheral blood and the patient died of multiorgan failure eight months after presentation. The blasts were positive for T cell markers (CD2, 4, 8) and NK marker (NKH-1) as well as monocytic markers. This finding, together with those by other investigators, may indicate the association between these immunophenotypes and extramedullary manifestations of acute monocytic leukemia.


[Hairy cell leukemia successfully treated with deoxycoformycin].

A 45-year-old male was hospitalized on September 2, 1989 with chief complaints of general fatigue and fever. Physical examination revealed hepatomegaly and massive splenomegaly. Laboratory tests on admission showed Hb of 7.5g/dl, PLT 4.8 x 10(4)/microliters and WBC 9,610/microliters with 81% hairy cells. Bone marrow aspirate demonstrated 55.1% hairy cells and moderate myelofibrosis. Cytochemically, hairy cells were positive for tartrate-resistant acid phosphatase (TRAP). Surface markers were SmIg G+ A+ kappa +, CD11b+, CD11c+, CD19+, CD20+, CD21-, CD25+, HC2+, HLA-DR+. From these findings, a diagnosis of hairy cell leukemia (HCL) was made. After administration of deoxycoformycin (DCF) at a dose of 5.0mg/m2 1-2 times monthly, splenomegaly disappeared, as did hairy cells from the peripheral blood. Hematological level returned to within normal range except for the presence of 1.2% hairy cells and mild myelofibrosis in bone marrow aspirates. DCF has so far been effective for this patient. While DCF has been reported to be effective in the treatment of HCL in the West, it has not been determined in Japanese patients with HCL, who have different hematologic features from those of HCL patients in the West.


[Iron deficiency anemia and hypoproteinemia induced by fresh cow's milk].

We reported two patients who developed severe iron deficiency anemia and hypoproteinemia which were induced by excessive drinking cow's milk. Case 1, a 23-month-old girl, had started drinking 800-1,000ml of cow's milk everyday since the age of 9 months. She developed generalized edema at the age of 23 months and visited Saga Medical School for Treatment. Case 2, a 14-month-old girl began to drink cow's milk at the age of 6 months. She drank 1,500ml cow's milk every day at the age of 14 months. She was, referred to our hospital by her physician because of severe anemia. Hypoproteinemia and severe iron deficiency anemia in both of these patients were improved by oral iron intake and prohibiting of cow's milk. Although many children at their age usually drink more than 500ml cow's milk every day, manifestations of symptoms such as iron deficiency anemia and hypoproteinemia do not occur frequently. Thus some investigations suggest that genetic defects as well as other factors related to this disease may be important factors.


[Successful treatment with combination of methylprednisolone, antilymphocyte globulin and cyclosporine in two cases of aplastic anemia].

Two children with severe and moderate aplastic anemia received combination treatment with methylprednisolone, antilymphocyte globulin and cyclosporine. One child (severe type) was given 2 courses of high dose methylprednisolone during the last 4 weeks without improvement prior to this combination therapy. The other child (moderate type) was not previously treated. Both patients showed complete response within 3 months following this therapy. Serious side effects were not observed. Combination therapy of these three drugs may be beneficial for patients with aplastic anemia.


[Primary myelofibrosis successfully treated with allogeneic bone marrow transplantation].

A 31-year-old man with primary myelofibrosis initially received low dose Ara C. Splenomegaly decreased but pancytopenia continued. Allogeneic bone marrow transplantation from his sister was then performed. Busulfan and cyclophosphamide were used as a preconditioning regimen, which included neither irradiation nor splenectomy. As the bone marrow was hypoplastic after transplantation, G-CSF was given. It was useful for systemic infection. After transplantation, leukoerythroblastosis and tear drop poikilocytosis disappeared in peripheral blood. Finally, bone marrow fibrosis disappeared and hemopoiesis to normal limits recovered 17 months later. These results demonstrate that bone marrow transplantation is effective for primary myelofibrosis for which there is no otherwise curative therapy.


[Primary macroglobulinemia with hypercalcemia, renal failure and systemic amyloidosis].

This report deals with an unusual case of primary macroglobulinemia with hypercalcemia, chronic renal failure and systemic amyloidosis. In May 1990, a 63-year-old male was transferred to our hospital because of hypercalcemia (13.5 mg/dl) and renal failure. Clinical examinations showed anemia, macroglossia, lymph node swellings and hepatomegaly. Laboratory findings included Bence-Jones (kappa type) proteinuria (0.8 g/day), a monoclonal gammopathy of the IgM-kappa type (2.8 g/dl), a proliferation of lymphoid cells in the peripheral blood (5%) and the bone marrow (59.6%), and lymphomatous involvement of an inguinal lymph node. Serum creatinine concentration was 8.5 mg/dl. The serum levels of parathormone and vitamin D3 metabolites were normal. The roentgenogram of bones showed a compression fracture of the lumbar spine and systemic osteoporosis. The treatment included eel calcitonin, prednisolone and the CHOP regimen, followed by hemodialysis and plasmapheresis. The serum level of IgM increased to 4.6 g/dl. The patient died three months later and postmortem examination demonstrated marked systemic amyloidosis.


[Vincristine played a pivotal role in hematological remission of thrombotic thrombocytopenic purpura (T.T.P.)].

A case of thrombotic thrombocytopenic purpura (TTP), in which vincristine (VCR) administration played a pivotal role in achieving the hematological remission, is presented. A 40-year-old woman with a chief complaint of purpura was transferred for detailed examination of thrombocytopenia. The presence of hemolytic anemia, schistocytosis, febrile clinical course, renal dysfunction and neurological abnormalities made the basis of a diagnosis of TTP. Twelve courses of consecutive plasma exchange and plasma infusion, which amounted to a total of 120l of fresh frozen plasma, were performed without obvious hematological improvement. Therefore, VCR (2 mg) was infused in addition to the plasma exchange. Three days later, platelets responded remarkably. After 10 days of hematological improvement, relapse occurred. VCR was reinfused. However, the second VCR infusion alone was not able to elevate platelets. With concomitant use of significant amounts of plasma, the platelet count responded gradually. Hematological remission was obtained and has continued until now. The exact role of VCR for TTP is still unknown; however, VCR is suggested to be effective for TTP as in the case presented here.


[Chronic cold agglutinin disease terminating in primary macroglobulinemia after a 10 year history].

A 60-year-old man was admitted with general fatigue and jaundice of one year's duration in February, 1981. The hemoglobin (Hb) was 11.4 g/dl and reticulocytes were 1.7%. A diagnosis of chronic cold agglutinin disease (CCAD) was made from the presence of cold agglutinin (CA) 1:2,048, increased serum IgM 267 mg/dl and indirect bilirubin 1.4 mg/dl. His Hb was approximately 11 g/dl in summer and 9 g/dl in winter for the subsequent ten years without therapy. In July, 1990, he was readmitted because of exacerbation of anemia and hepatosplenomegaly. The Hb was 4.6 g/dl, indirect bilirubin 3. 1 mg/dl, CA titer 1:232,144 and reticulocytes were 20%. Serum IgM was 1,065 mg/dl, and immunoelectrophoresis showed IgM-kappa M-protein. Peripheral blood lymphoid cells expressed surface membrane immunoglobulin (SmIg) M and kappa. The bone marrow showed an increased number of lymphoid cells which also expressed SmIg M and kappa. These findings were compatible with those of the features of primary macroglobulinemia (PMG). The M-2 protocol resulted in decrease in serum IgM and CA, but he died of heart failure in February, 1991. The relationship between CCAD and PMG in relation to the pathogenesis was discussed.


[Two cases of acute promyelocytic leukemia in pregnancy and the effect of anthracyclines on fetal development].

Two patients with acute promyelocytic leukemia (APL) in 2nd and 3rd trimester of pregnancy are reported on. Case 1: 38-year-old female consulted our hospital because of bleeding tendency and pancytopenia in April, 1988. She was diagnosed as having APL with disseminated intravascular coagulopathy (DIC) and was found to be in the 14th week of gestation. Combined chemotherapy (BHAC-DMP) including the total dose (440 mg) of daunorubicin (DNR) resulted in intrauterine fetal death at 19 weeks of gestation. The fetus was severely anemic and the bone marrow was hypoplastic. Case 2: A 27-year-old female was diagnosed as having APL with DIC at 29 weeks of gestation. BHAC-DMP including 440 mg DNR achieved complete remission. At 35 weeks of gestation, she delivered a normal infant by Caesarean section. The child had normal hematological findings and showed normal growth. Both cases developed APL accompanied by DIC during pregnancy and were treated with a similar regimen including high dose of anthracyclines. Case 2 treated in the late period of gestation delivered a normal infant, while fetal death resulted in case 1, treated in the early period of gestation. We reviewed the literature regarding chemotherapy using anthracyclines during pregnancy.


[Multiple myeloma with a mass formation in a pacemaker pocket].

A 74-year-old female was diagnosed as having multiple myeloma in August 1989 and was treated with combined vincristine, melphalan and prednisolone. Subsequently, she was followed followed up in the outpatient clinic using interferon-alpha. On August 6, 1990, she had a transvenous demand pacemaker inserted because of severe atrioventricular block. The pulse generator was placed in a subcutaneous pocket in the left pectoral area. On February 3, 1991, she developed a mass overlying the pulse generator. This tumor was diagnosed as plasmacytoma by histological examinations. A myelogram showed 5.1% plasma cells with 5.5 x 10(4) nucleated cells/microliter. The amounts of serum protein and IgA M protein were 6.8 g/dl and 1.8 g/dl, respectively. The tumor responded to combined chemotherapy, but reenlarged to the initial size 3-4 weeks later. On August 6, 1991, this tumor, including the pulse generator was removed. By October 1991, the patient had systemic subcutaneous tumors and a right maxillary tumor suggesting the aggressive phase. On December 19, 1991, she died due to cardiac failure. In this paper the discussion focussed on the etiopathogenesis of plasmacytoma arising in the region of pulse generator pockets.


Common phenotypic expression of gangliosides GM3 and GD3 in normal human tissues and neoplastic skin lesions.

The expression of gangliosides in non-malignant tissues (epidermis and pigmented nevus) and neoplastic lesions (melanoma, squamous cell carcinoma [SCC] and basal cell carcinoma [BCS]) of the human skin was analyzed immunohistochemically and biochemically to characterize the features associated with malignancy. Immunohistochemical staining with an anti-II3NeuAc-LacCer (GM3) monoclonal antibody (M2590 mAb) and an anti-II3(NeuAc)2-LacCer (GD3) mAb (R24) showed the expression of the gangliosides GM3 and GD3 to vary among the different tissues. M2590 clearly stained epidermal keratinocytes and the tumor cells of BCC and SCC, and strongly stained melanocytes and melanoma cells. In contrast, R24 did not stain epidermal keratinocytes and only faintly stained SCC cells, while it clearly stained BCC cells, and intensely stained melanocytes and melanoma cells. GM3 showed a similar level of staining among the tissue specimens, while the level of GD3 staining was quite variable among the tumor specimens. Biochemical analysis by thin-layer chromatography (TLC) with resorcinol staining and TLC immunostaining with either M2590 or R24 showed both GM3 and GD3 to be commonly expressed by both the normal and malignant skin tissues, including SCC. There was no close correlation between the intensity of immunohistochemical staining and the biochemically detected amounts of these gangliosides. This may have been partly due to the so-called cryptic expression of cell membrane gangliosides. Our results thus suggest that analysis of the tumor-associated expression of gangliosides requires several methods, since the sensitivity of the methods used may have a considerable effect on the diagnostic value of gangliosides as skin cancer markers.


Urinary excretion of parathyroid hormone-related protein as a predictor of hypercalcemia in patients with adult T-cell leukemia.

Hypercalcemia with adult T-cell leukemia (ATL) is chiefly caused by an excessive production by tumor cells of parathyroid hormone-related protein (PTHrP). We have previously reported hypercalcemic patients with solid tumors to excrete a large amount of the C-terminal fragments of PTHrP (C-PTHrP) into their urine. To elucidate whether PTHrP production correlates with or predicts the development of hypercalcemia, we studied the urinary excretion of C-PTHrP in 36 ATL patients. The urinary excretion of C-PTHrP was in the normal range (< 0.40 nmol equivalent to PTHrP (109-141)/g creatinine) in HTLV-1-positive carriers (n 3), ATL patients in complete remission (n 2) and chronic type ATL patients (n 2). It was marginally increased in seven patients in partial remission, and gradually increased as the disease progressed. In 20 patients who died without or with hypercalcemia, it was increased to 1.98 +/- 0.69 (n 9) and 7.6 +/- 2.1 nmol/g creatinine (mean +/- SD, n 11, P < 0.01), respectively. Urinary C-PTHrP excretion was significantly correlated with serum calcium and LDH levels as well as with CD25-positive cells in the peripheral blood. In four patients whose urinary excretion had been serially determined, it increased prior to the development of hypercalcemia. The findings suggest the urinary excretion of C-PTHrP to be of use as a predictor of the development of hypercalcemia in ATL patients. In ATL patients whose urinary excretion of C-PTHrP is progressively increasing, the serum calcium concentration should be carefully monitored to prevent hypercalcemic crisis.


Surgical treatment of adrenal diseases at Nara Medical University and affiliated hospitals.

The effect of advanced diagnostic imaging on the surgical treatment of adrenal diseases was examined in 86 cases of adrenal disease surgically treated at the Department of Urology, Nara Medical University, and affiliated hospitals from August, 1962, to July, 1991. The number of cases included in the present study was 16 for the period before the introduction of CT (from 1962 to 1978), and 70, including 13 cases of adrenal incidental tumor, after the introduction of CT in 1979. There were no malignant tumors histopathologically, among the 13 cases of incidental tumor. Accurate localization of functioning adrenal disease was possible at a rate of 100% with angiography, including selective venous blood sampling, 98.6% with CT and 84.5% with ultrasonography. The rate was 100% with magnetic resonance imaging which was carried out on 19 cases (20 adrenal glands). Signs of malignancy were observed in the removed tissue of three cases from the pheochromocytoma group. As the diagnostic imaging technique has improved, the use of a flank approach has increased in the surgery for adrenal disease.


Prognosis of osteosarcoma treated by limb-salvage surgery: the ten-year intergroup study in Japan.

We report the prognoses for 107 patients treated by limb-salvage surgery and 147 treated by amputation or disarticulation, during the period 1980-1985, at 22 university hospitals, cancer centers and national hospitals in Japan. In this multi-institute non-randomized study, the five-year cumulative survival rate was 70% for the former group and 49% for the latter. The prognosis was more favorable among patients who responded to preoperative chemotherapy than among those who did not. Limb-salvage surgery was feasible for most of the surgical stage IIB patients with small extraosseous tumor extensions who had responded to preoperative chemotherapy. Local tumor recurrences were seen in 15 (14%) of the 107 patients in the limb-salvage surgery group, nine of whom died of the metastasis. While limb-salvage surgery is being increasingly used in Japan, present indications show the procedure not to reduce survival rate for osteosarcoma patients. The survival rate of patients with wide-with-marginal margins was similar to that of patients with wide margins. Taking into account the long-term survival of osteosarcoma patients, limb-salvage surgery can be recommended when the osteosarcoma responds to preoperative chemotherapy.


Estimating a summarized odds ratio whilst eliminating publication bias in meta-analysis.

Publication bias is a recognized phenomenon, i.e. studies with statistically significant results are more likely to be published than those finding no difference between the groups studied. Summarized odds ratio calculated from odds ratios of published studies in a meta-analysis may be overestimated because of publication bias. This is a significant problem in research areas involving weak associations between causes and results. The magnitude of publication bias in a given research area cannot be determined directly. The present study enables us to calculate the summarized odds ratio of hypothetical unpublished studies from odds ratios of published studies indirectly, employing a moment method by assuming the natural logarithm value of the odds ratio to be distributed normally. We can then estimate summarized odds ratio in all studies, which include not only those published but also those unpublished. When these studies are homogeneous in quality and their odds ratios homogeneous in quantity, the method can eliminate publication bias.


An ileal neobladder for a female patient after a radical cystectomy to ensure voiding from the urethra: a case report.

An ileal neobladder construction realizing normal micturition was successfully performed after a total cystectomy, with preservation of the urethra, for a 69-year-old female patient with invasive bladder cancer. Approximately 60 cm of terminal ileum was selected, detubularized and re-sutured to create an oval-shaped intestinal pouch to which the bilateral ureters were anastomosed. The bottom of the pouch was anastomosed to the urethral remnant. Three months after surgery, the patient achieved daytime continence, and now enjoys almost the same lifestyle as before. This is the first case of successful neobladder construction to be precisely reported for a female patient. For normal micturition, the ileal neobladder will be one of the possible choices for urinary diversion in not only selected male, but also female, patients.


Supraventricular tachycardia in an infant.

Treating this 1-month-old infant with SVT with adenosine was extremely effective and resulted in no observable side effects to the patient. More research needs to be performed and documented regarding the effectiveness and safety of this drug in pediatric patients. Emergency nurses should familiarize themselves with adenosine and should suggest its use in pediatric patients with SVT that does not respond to vagal maneuvers.


Complete heart block in the pediatric patient.

Care for the pediatric patient with CHB focuses on signs and symptoms associated with respiratory failure and shock. Differences in size among pediatric patients--and in anatomy, physiology, and possible causes--should be considered. In children, cardiac arrest is usually secondary to respiratory arrest. Respiratory arrest should be anticipated in a pediatric patient with (1) decreased level of consciousness, (2) poor muscle tone, (3) increased respiratory rate or respiratory effort (nasal flaring, intercostal, subcostal, and suprasternal retractions); or (4) cyanosis. Shock is the failure of the cardiovascular system to perfuse vital organs adequately. Organ perfusion is determined by cardiac output, which in turn results from the heart rate times stroke volume. In CHB the slowing of heart rate can significantly decrease cardiac output, so the signs and symptoms of shock--especially level of consciousness, peripheral circulation, and blood pressure--must continually be reassessed.


Abuse, neglect, abandonment, violence, and exploitation: an analysis of all elderly patients seen in one emergency department during a six-month period.

Vulnerable individuals in society seem to be even more vulnerable as they age. Our data suggest that during the next decade members of our aging society may well tax our adult protective service systems to a greater degree than heretofore imagined. We have an opportunity to be proactive in this regard.


Specialized services for psychiatric patients in the emergency department.

The care of the psychiatric patient in the emergency department offers yet another opportunity for emergency nurses to draw on specialty nursing resources or to create expanded roles for emergency practitioners themselves. As our emergency departments continue to encounter increasing numbers of patients, the need for concise and expeditious psychiatric consultation protocols becomes even more clear.


Triage assessment tool.

The Triage Assessment Tool, although not meant to replace clinical judgment and experience, is designed to augment the decision making skills of the triage nurse. This is attained by focusing the user on the importance of sequence, and examination of pertinent data, thus ensuring a precise triage decision.


[Local corticosteroid therapy in childhood].

Various dermatoses require the application of topical glucocorticoids (gc) already in childhood. There are mostly inflammatory and frequently chronic skin diseases. In necessary long-term therapy the risk of topical and/or systemic side effects increases. This depends among others on special features of the children's skin, the potency of the gc and the mode of application. To avoid side effects, variations of a discontinuous therapy and so called weak gc should be used.


[The kinetics of vaccine antibodies against tetanus toxoid, diphtheria toxoid, measles virus, poliomyelitis virus and pneumococcus after allogeneic and autologous bone marrow transplantation and revaccination. 3: The kinetics of vaccine antibodies against tetanus toxoid and diphtheria toxoid after allogeneic and autologous bone marrow transplantation and combined revaccination against diphtheria and tetanus].

The 3rd part of the paper deals with the results of a combined revaccination against diphtheria and tetanus in a group of 25 children after allogeneic bone marrow transplantation (BMT) with and without graft versus host disease (GvHD) and after autologous transplantation. It can be shown that in the allogeneic transplanted groups with and without GvHD it is possible to build up a tetanus and diphtheria antitoxin titre in a safe protective cause by a 2nd basic immunisation consisting of 3 single vaccinations starting about 9 to 12 months later. For autologous transplanted children only 1 to 2 vaccinations at a later term than for the allogeneic transplanted children may possibly be sufficient.


[Hypohidrotic ectodermal dysplasia as the cause of recurrent hyperthermia in a young infant].

Fever--the most frequent symptom of diseases in children--points diagnostically to an infection. We report on a child who presented with recurrent high fever of unknown origin. Infectious diseases could be excluded. During a general medical checkup the results of repeated pilocarpine iontophoresis attracted attention by the minimal secretion of sweat. The diagnosis of hypohydrotic ectodermal dysplasia could be confirmed by dermal biopsy since all integumentary appendages were missing. Only 300 cases of this rare hereditary dysplasia can be found in the literature.


[The renal effects of dopamine therapy in premature infants in the first week of life].

It is known that Dopamine therapy induces loss of water and sodium on the first or second days of life. On the other hand, no studies have been published on renal function under long-term therapy. In our study we could prove an altered excretion of water and sodium as well as fractional sodium excretion in preterm infants who received dopamine for at least the first week of life. Conclusions for clinical practice are drawn.


[Laser angioplasty in the staged surgical treatment of patients with combined and multiple lesions of different arterial basins].

Percutaneous laser angioplasty was attempted in 30 patients with occlusive lesions of the main arteries of the lower extremities and the branches of the aortic arch. Most patients had either combined affection of the branches of the aortic arch and arteries of the lower extremities (56%), or involvement of several vascular segments on one extremity into the process (74%), or involvement of arteries of both lower extremities (85%). Seven patients underwent stage correction of circulatory cerebral disorders and restoration of patency of arteries of both lower extremities. Excimer laser on a wave length of 308 nm was used for angioplasty. Adequate operations were carried out in all cases without any complications in the postoperative period.


[Immunological disorders of homeostasis and the methods of their correction for the programmed and preoperative treatment of patients with nonspecific aorto-arteritis].

The authors examined 29 patients with aorto-arteritis of unspecific origin of different duration and variants of affections of the major vessels. The patients were divided into groups with active and inactive course of the inflammatory process. The changes of immunological values and the phagocytosis data were more marked in the group with the active phase of the disease. These changes correlated with the data on unspecific inflammation and the clinical picture. Twenty-two patients were treated by hemosorption and immunocorrective measures. The fibrinogen concentration reduced, the ESR was normalized, and C-reactive protein became negative. The concentration of circulating immune complex and immunoglobulins diminished, the phagocytosis values became close to normal. It is concluded that hemosorption is an effective method of correction of immunological disorders in unspecific aorto-arteritis, both as an independent therapeutic method and as a measure of preoperative management.


[Atherosclerotic aneurysm of the iliac arteries].

Operations were performed on 5 patients with isolated atherosclerotic aneurysms of the iliac arteries, who accounted for 10.9% of patients with aneurysms of vessels treated at the Vascular Department of the Novgorod Regional Hospital from 1983 to 1990. In 4 patients the aneurysm of the iliac artery was an operative finding, in one patient it was found only on postmortem examination. The diagnostic errors and causes of a fatal outcome in 2 patients are discussed. The authors claim resection of the aneurysm with reconstruction of the aorto-ilio-femoral segment to be the operation of choice in aneurysm of the iliac artery.


[The assessment of the efficacy of surgical treatment in critical ischemia of the lower extremities by the forming of arteriovenous anastomoses].

The authors show the significance of radionuclide angiography in evaluating the efficacy of surgical treatment of lower limb critical ischemia by formation of an arteriovenous anastomosis on the side of the affection. An operation for formation of an arteriovenous anastomosis on the lower third of the leg or the foot was found to be most effective. Increased perfusion of the ischemic limb on the level of balanced mixing of the blood flow was revealed in most patients, which the authors believe to be caused by venous stasis in the lower limb due to reduced difference in pressure in the arterial and venous bed. The highest percentage of favorable late--term results of the operation was revealed among this contingent of patients.


[Vasotonometry in the episcleral arteries in the diagnosis of stenotic lesions of the extracranial sections of the carotid arteries].

The object of the work was evaluation of the method of vasotonometry (VTM) in the episcleral arteries in recognizing stenoses of the extracranial part of the internal carotid artery (ICA). The examination was conducted by means of a specially designed vasotonometer in patients suffering from ischemic type cerebral circulatory disorders. The authors examined 98 patients and 20 healthy volunteers. The parameters of the diagnostic sensitivity of the method in various degrees and forms of the stenotic lesions of the ICA were determined. The correct diagnosis of a hemodynamically significant ICA stenosis was established in 85% of patients. The results of the examination were confirmed by angiography and operation.


[Surgery of the major vessels in cancer patients].

The article generalizes 25-year experience in various vascular surgical interventions conducted on 159 oncological patients. Planned operations were conducted after thorough angiographic study in 42 patients (arteriolysis and phlebolysis in 11, thrombectomy in 13, plastics of the vessels in 18). Two patients died after thrombectomy. Embolization of the renal arteries was undertaken in 55 patients with renal carcinoma; there were no fatal outcomes. Various operations for iatrogenic injury to the vessels were carried out in 60 patients (lateral suture in 28, circular suture in 3, ligation of vessels in 18). In this group 11 patients (18.3%) died. The authors claim that large oncological centers should be equipped with the appropriate angiographic apparatuses and have trained angio-surgeons on their staff.


[The role of thermography in the diagnosis of obliterating vascular diseases of the lower extremities].

The authors undertook comparative evaluation of the various methods of diagnosis of obliterative diseases of the lower limb vessels in 32 patients with II-III stage arteriosclerosis obliterans. Comparison of the results of arteriography, ultrasonic dopplerography, and objective examination with those of thermovision examination led to the conclusion that the method is valuable in revealing the condition of the collateral bed and microcirculation and the compensatory mechanism of circulation on the whole. In similar treatment in groups of patients formed according to the level of obliteration of the vessels and the thermographic picture, the best results were produced in those groups in which the intensity of the infrared rays was higher, which allowed the conclusion that thermography possesses not only diagnostic but also prognostic significance.


[The transfascial excision of indurated subcutaneous fat in the postthrombophlebitic syndrome].

The authors came to the conclusion that in surgical treatment of the postthrombophlebitic syndrome it is necessary in some cases to excise the indurated fatty tissue on the leg during the operation on the veins. It should be carried out transfascially through a Felder incision in combination with ligatin of the perforating veins on the leg. The suggested method produces better results than those of excision of the changed fatty tissue through a cut made in the skin overlying it. Excision of the indurated fatty tissue removes the focus of chronic infection and autoallergy.


[The pathogenesis of acute thrombophlebitis of the lower extremities].

It is emphasized in the discussed problem that several factors participate simultaneously in the mechanism of thrombogenesis. At the same time, local hemodynamic disorders (congestion, turbulent flow, local hypoxemia) lead to pathological changes of blood platelet hemostatic properties in these regions, which is conducive to local thrombus formation. Inflammation develops as a consequence of the response of the venous wall to stimulators of infectious, autoimmune or neoplastic character arriving along the lymphatic vessels, capillaries and perivascular spaces.


[The role of functional phlebography in the diagnosis of chronic postphlebitic venous insufficiency of the lower extremities].

The work deals with the description of methods for phlebography of the lower extremities with the patient in a vertical pos position with the use of various functional, loading, and pharmacological tests. Such examination makes it possible to recognize the pathological changes, reveal precisely the architectonics of the venous system and the localization of the most changed areas of the venous system, and determine the rate of the venous blood flow. Much attention is paid to preventive measures during phlebographic examination. The efficacy of the phlebography method is proved on a rich clinical material and the indications and contraindications for it are specified.