Prolonged intravenous urokinase infusion: an alternative pharmacologic approach in the treatment of thrombus-containing saphenous vein graft stenoses.

A subtotally occlusive saphenous vein graft stenosis resolved after rapid intracoronary and prolonged intravenous urokinase infusion. Additional therapy was unnecessary, avoiding the attendant risks of saphenous vein graft angioplasty. Combined intracoronary and intravenous urokinase infusion should be considered prior to coronary angioplasty of saphenous vein graft stenoses, particularly when diffuse degeneration is present or the risk of underlying thrombus is high.


New approach to management of intraaortic balloon pumps in patients with peripheral vascular disease: case reports of four patients requiring urgent IABP insertion.

Four selected cases of emergent IABP insertion in PV patients are presented. After angiographic documentation of critical iliac stenosis, conservative peripheral angioplasty was performed prior to IABP insertion. No patient experienced a peripheral ischemic event associated with IABP use.


Antegrade selective coronary angiography via the transseptal approach in a patient with severe vascular disease.

We describe a case of a woman with severe vascular disease in whom retrograde access to the aortic root was limited by both aortoiliac and axillary disease. Transseptal catheterization was performed in anticipation of percutaneous aortic valvuloplasty. Selective antegrade angiography was successfully performed using catheters introduced through the transseptal sheath.


Pseudoaneurysm of coronary artery following rupture of coronary artery during coronary angioplasty.

Rupture of the coronary artery is a rare complication of percutaneous transluminal coronary angioplasty (PTCA). We describe a case of coronary artery rupture during PTCA resulting in the formation of a coronary artery pseudoaneurysm. The pseudoaneurysm was successfully treated by percutaneous spring-coil embolization of the coronary artery.


Interpretation of cardiac pathophysiology from pressure waveform analysis: mitral valve gradients: Part I.

The mitral valve gradient is dependent on the precise measurement of left atrial (or pulmonary capillary wedge) and left ventricular pressures. Artifacts involving either pressure measurement will produce inaccuracies which may have clinical significance. Several methods and formulas using both invasive and noninvasive techniques should verify clinical findings and confirm the severity of mitral valve disease prior to definite therapy. The changes in mitral valve gradients after balloon catheter valvuloplasty will be discussed in part II of this hemodynamic rounds.


Percutaneous vascular hemostasis device for interventional procedures.

A new 11.5F over-the-wire vascular hemostasis device was compared to conventional manual compression in normal swine femoral arteries. Using percutaneous techniques, the collagen was deposited and hemostasis achieved in 7 vessels after 1 minute total, plus 4 minutes partial compression, while control manual compression required more than 5 minutes total compression to avoid hematoma formation. One month follow-up of treated arteries (n = 4) and controls (n = 3) showed no impairment in distal pulse or differences in histology at the puncture site in the control and treated arteries. Thus, the vascular hemostasis device technique is safe and effective, achieves hemostasis with less compression time and complications, and does not interfere with arterial wall healing or compromise the lumen.


Inoue balloon usefulness in case of failure to stabilize bifoil catheter balloons during percutaneous mitral valvotomy.

A new bifoil balloon catheter has been used in the last 120 percutaneous mitral valvotomies carried out in our institution. The shaft segment between the two balloons of the bifoil catheter has been adjusted to the guide-wire diameter, allowing its introduction through a 14F sheath. This thinner shaft does not always offer enough back up to the balloons during inflation. An Inoue balloon replaced an unstable bifoil balloon in 5 cases of mitral dilatation failure due to balloon instability, regularly providing firm stability until full balloon inflation.


The structure of human mitochondrial manganese superoxide dismutase reveals a novel tetrameric interface of two 4-helix bundles.

The 2.2 A resolution crystal structure of recombinant human manganese superoxide dismutase, a homotetrameric enzyme that protects mitochondria against oxygen-mediated free radical damage, has been determined. Within each subunit, both the N-terminal helical hairpin and C-terminal alpha/beta domains contribute ligands to the catalytic manganese site. Two identical 4-helix bundles, symmetrically assembled from the N-terminal helical hairpins, form novel tetrameric interfaces that stabilize the active sites. Structurally altered polymorphic variants with reduced activity, such as tetrameric interface mutant Ile-58 to Thr, may produce not only an early selective advantage, through enhanced cytotoxicity of tumor necrosis factor for virus-infected cells, but also detrimental effects from increased mitochondrial oxidative damage, contributing to degenerative conditions, including diabetes, aging, and Parkinson's and Alzheimer's diseases.


A nucleosome core is transferred out of the path of a transcribing polymerase.

We have determined the fate of a nucleosome core on transcription. A nucleosome core was assembled on a short DNA fragment and ligated into a plasmid containing a promoter and terminators for SP6 RNA polymerase. The nucleosome core was stable in the absence of transcription. The distribution of nucleosome cores after transcription was examined. The histone octamer was displaced from its original site and reformed a nucleosome core at a new site within the same plasmid molecule, with some preference for the untranscribed region behind the promoter. These observations eliminate several models that have been proposed for transcription through a nucleosome core. Our results suggest that a nucleosome core in the path of a transcribing polymerase is displaced by transfer to the closest acceptor DNA.


A soluble receptor for interleukin-1 beta encoded by vaccinia virus: a novel mechanism of virus modulation of the host response to infection.

Vaccinia virus gene B15R is shown to encode an abundant, secretory glycoprotein that functions as a soluble interleukin-1 (IL-1) receptor. This IL-1 receptor has novel specificity since, in contrast with cellular counterparts, it binds only IL-1 beta and not IL-1 alpha or the natural competitor IL-1 receptor antagonist. The vaccinia IL-1 beta receptor is secreted when expressed in a baculovirus system and competitively inhibited binding of IL-1 beta to the natural receptor on T cells. Deletion of B15R from vaccinia virus accelerated the appearance of symptoms of illness and mortality in intranasally infected mice, suggesting that the blockade of IL-1 beta by vaccinia virus can diminish the systemic acute phase response to infection and modulate the severity of the disease. The IL-1 beta binding activity is present in other orthopoxviruses.


Molecular basis of human hypertension: role of angiotensinogen.

Essential hypertension is a common human disease believed to result from the interplay of multiple genetic and environmental determinants. In genetic studies of two large panels of hypertensive sibships from widely separated geographical areas, we obtained evidence of genetic linkage between the angiotensinogen gene (AGT) and hypertension, demonstrated association of AGT molecular variants with the disease, and found significant differences in plasma concentrations of angiotensinogen among hypertensive subjects with different AGT genotypes. The corroboration and replication afforded by these results support the interpretation that molecular variants of AGT constitute inherited predispositions to essential hypertension in humans.


Secretion and localized transcription suggest a role in positional signaling for products of the segmentation gene hedgehog.

The segment polarity genes engrailed and wingless are expressed in neighboring stripes of cells on opposite sides of the Drosophila parasegment boundary. Each gene is mutually required for maintenance of the other's expression; continued expression of both also requires several other segment polarity genes. We show here that one such gene, hedgehog, encodes a protein targeted to the secretory pathway and is expressed coincidently with engrailed in embryos and in imaginal discs; maintenance of the hedgehog expression pattern is itself dependent upon other segment polarity genes including engrailed and wingless. Expression of hedgehog thus functions in, and is sensitive to, positional signaling. These properties are consistent with the non-cell autonomous requirement for hedgehog in cuticular patterning and in maintenance of wingless expression.


slow border cells, a locus required for a developmentally regulated cell migration during oogenesis, encodes Drosophila C/EBP.

During Drosophila oogenesis six to ten follicle cells, the border cells, undergo a dramatic and stereotypic migration through the developing egg chamber. We identified four independent P element insertion mutations that specifically blocked border cell migration. They defined a single, novel locus that was named slow border cells (slbo), because hypomorphic alleles caused delayed onset of the migration. Laser ablation of the border cells, or failure of their migration, caused improper morphogenesis of the micropyle, the egg-shell structure through which the sperm enters at fertilization. The slbo locus was found to encode a product homologous to the CCAAT/enhancer-binding protein (C/EBP), a basic region-leucine zipper transcription factor. Drosophila C/EBP may be required for the expression of gene products mediating border cell migration.


Bidirectional movement of a nascent polypeptide across microsomal membranes reveals requirements for vectorial translocation of proteins.

The translocation of polypeptides across the endoplasmic reticulum is a vectorial process that occurs probably through a protein channel by a mechanism as yet undetermined. Here, we demonstrate bidirectional movement of a 221 residue nascent polypeptide across microsomal membranes and provide evidence suggesting that the retrograde movement is through the translocation channel. Retrograde movement is observed only when the polypeptide is generated from a truncated transcript; addition of a stop codon after codon 221 confers vectorial movement. Retrograde movement can also be prevented by glycosylation of the nascent polypeptide, as well as by inclusion of 32 additional amino acids that may promote folding of the translocated chain. We propose that the protein translocation channel is a passive pore that does not create a directional bias in polypeptide movement and that vectorial translocation is driven by nascent chain elongation and sustained by posttranslocation events that prevent retrograde movement.


The translation machinery and 70 kd heat shock protein cooperate in protein synthesis.

The function of the yeast SSB 70 kd heatshock proteins (hsp70s) was investigated by a variety of approaches. The SSB hsp70s (Ssb1/2p) are associated with translating ribosomes. This association is disrupted by puromycin, suggesting that Ssb1/2p may bind directly to the nascent polypeptide. Mutant ssb1 ssb2 strains grow slowly, contain a low number of translating ribosomes, and are hypersensitive to several inhibitors of protein synthesis. The slow growth phenotype of ssb1 ssb2 mutants is suppressed by increased copy number of a gene encoding a novel translation elongation factor 1 alpha (EF-1 alpha)-like protein. We suggest that cytosolic hsp70 aids in the passage of the nascent polypeptide chain through the ribosome in a manner analogous to the role played by organelle-localized hsp70 in the transport of proteins across membranes.


Effects of glutathione on the synthesis and turnover of interleukin-2 receptors.

Internalization of IL-2 is important for its biological activities. The internalization of IL-2 was regulated by the duration of glutathione (GSH) treatment in CTLL-2 and CT-4R cells. Flow cytometric studies showed that the level of surface IL-2 receptors was not increased by GSH treatment. Northern blot analysis showed that the mRNA of IL-2Rp55 and IL-2Rp70, the two major components of the high-affinity IL-2 receptors, was increased 6 hr after GSH treatment. The appearance rate of membrane IL-2 receptors in GSH-treated cells was faster than that of the untreated cells. GSH also shortened the half-life (from 5 to less than or equal to 3 hr) and thus increased the turnover of the surface high-affinity IL-2 receptors. These results suggest that although GSH does not affect the level of surface IL-2 receptors, GSH may regulate the internalization of IL-2 by enhancing the synthesis and turnover of surface IL-2 receptors.


Agonistic effects of tyrphostins on human peripheral mononuclear cells.

Tyrosine kinases of the src family, p56lck and p59fyn, were implicated in the transduction of signals via the T-cell receptor complex. These kinases are negatively regulated by phosphorylation of a carboxyl-terminal tyrosine residue. Tyrphostins are synthetic low molecular weight compounds that selectively inhibit different protein tyrosine kinases. We report here on the agonistic and antagonistic effects of tyrphostins on human peripheral blood mononuclear cells (PBM). At low concentration, the tyrphostins enhanced glucose uptake and maximal stimulation was attained at a concentration characteristic for each of the tyrphostins used. Higher concentrations were less effective. The tyrphostins AG126 and AG183 were also found to enhance IL-2-induced cytotoxicity in human PBM in a biphasic manner. In contrast, the tyrphostin AG17 markedly inhibited IL-2-induced cytotoxicity at low AG17 concentration and no stimulation was observed. The tyrphostins tested had selective effects on [3H]thymidine incorporation induced by the mixed lymphocyte culture and different agents. The most potent inhibitor was AG17. Tyrphostins also affect cytokine secretion by human PBM. AG126 and AG183 enhanced TNF-alpha secretion and this effect was more prominent in the presence of IL-2. AG126 enhanced IFN-gamma, IL-1, and IL-6 production in PBM that were costimulated with the stress stimuli heat shock and phenylarsine oxide. The stimulatory effects of the tyrphostins on cytokine secretion and induction of cytotoxicity might be interrelated. The agonistic and antagonistic effects of tyrphostins on lymphocyte functions may have therapeutic potential.


Estrogen accelerates immune complex glomerulonephritis but ameliorates T cell-mediated vasculitis and sialadenitis in autoimmune MRL lpr/lpr mice.

Estrogen is known to influence immune responses in healthy subjects in a dichotomous fashion. Thus, in number of previous studies we and others have demonstrated that B cell activities are augmented after exposure to estrogen whereas T cell reactivity is suppressed. Furthermore, it has been shown that this hormone has significant impact on the course of certain human and experimental autoimmune diseases. In this study we report that treatment with physiological doses of estradiol exerts dichotomous effects on different manifestations of the lupus disease in MRL/l mice. On one hand immune complex-mediated glomerulonephritis was significantly accelerated. This outcome was due to polyclonal B cell activation with increased production of antibodies to double-stranded DNA and formation of circulating immune complexes. In contrast, T cell-mediated lesions such as focal sialadenitis, renal vasculitis, and periarticular inflammation were all significantly ameliorated in MRL/l mice exposed to estrogen. Thus, we were able to demonstrate that, within one subject and even within one organ, administration of estrogen leads to differential outcome of SLE morbidity. We propose that the differential effect of estrogen on the manifestations of the autoimmune disease of MRL/l mice is due to its dichotomous effects on B and T cell-mediated immune responses.


The involvement of protein kinase C in activation-induced cell death in T-cell hybridoma.

T-cell hybridoma activated by a variety of stimuli such as anti-cell surface antigen, notably CD3 and T-cell receptors, and Con A undergoes a cell lysis process called activation-induced cell death (AICD). It was found that the major protein kinase C (PKC) isoform in the 2B4.11 T-cell hybridoma, PKC(alpha), was translocated from the cytosolic to the particulate fraction when these hybridoma cells were induced to die by plastic-adsorbed anti-CD3 antibodies. Inhibitors of protein phosphorylation rescued 2B4.11 cells from AICD as determined by the analysis of cellular metabolism and the proportion of living cells. Furthermore, PKC(alpha) down-regulation by phorbol ester treatment abolished AICD, and the degree of PKC down-regulation correlated well with the degree of AICD abolishment, suggesting that PKC activation represents an essential step in the molecular mechanisms underlying AICD in this T-cell hybridoma.


Mixed interleukins and thymosin fraction V synergistically induce T lymphocyte development in hydrocortisone-treated aged mice.

Analysis of the role of interleukins in T cell ontogeny in vitro indicates that the regulation of T cell development involves interleukins (ILs) as well as thymic hormones (THs). In order to assess their respective roles in T lymphocyte development in vivo, chemically thymectomized mice were treated with ILs and THs. After 2 days of hydrocortisone treatment, aged mice showed acute thymic involution (weight was less than 30% of control) and reduced spleen size (less than 80% of control) with progressive recovery to 8 days. After 2 days of hydrocortisone treatment, adult mice were injected for 5 days with mixed buffy coat interleukins (BC-IL; 50 units IL2 equivalence), purified IL2 (50 units), rIL1 beta (4 ng), and thymosin fraction V (TF5; 100 micrograms). The animals were sacrificed and spleens and thymuses were analyzed for weight, cellularity, T cell number, subsets, and function as determined by proliferative responses to concanavalin A and ILs. BC-IL treatment increased the recovery of spleen and thymus weights and cellularity with corresponding augmentation of number and function of T lymphocytes; neither IL1 or IL2 or their combination had this effect. TF5 had no effect alone but strongly potentiated the effect of BC-IL on T lymphocyte function. These data indicate that BC-IL in combination with thymic peptides potently promotes T lymphocyte development. The combination may be therapeutically relevant for immunorestoration.


Treatment of fetal thymic organ culture with IL-1 leads to accelerated differentiation of subsets of CD4-CD8- cells.

Using fetal thymic organ culture (FTOC), we describe the effects of IL-1 on T cell differentiation, particularly within the CD4-CD8- subset. While treatment of FTOC with IL-1 led to a modest reduction in total thymocyte yield, it induced an increase in the percentage of CD4-CD8- cells that express IL-2R early in culture and a decrease in the number of their precursors (CD44+IL-2R- cells). The increase in the percentage of cells expressing IL-2R was not accompanied by an increase in the number of these cells. At later time points these IL-2R+ cells (and their precursors) were reduced relative to controls. The total number of CD4-CD8-CD3- precursor cells in IL-1-treated cultures was reduced to approximately half that in controls at Day 12 of culture. However, only minor inhibition of total cell number was observed, which, taken together with the greater frequency of IL-2R+ precursors, suggests that this depletion of the pool of precursors may have been due to the induction of premature differentiation rather than to its inhibition.


Suppression of interleukin-2 and interleukin-2 receptor expression in Jurkat cells stably expressing the human immunodeficiency virus Tat protein.

The Jurkat T cell line was stably transfected with an Epstein-Barr virus-based episomal replicon designed to express high levels of the HIV-1 Tat protein. After selection in hygromycin B, high-level Tat activity was detected in 3 of 18 transfected cell lines. After stimulation with phytohemagglutinin (PHA) and phorbol myristate acetate (PMA), Tat transfectants with high Tat expression showed diminished expression of interleukin-2 (IL-2) and the interleukin-2 receptor alpha chain (IL-2R) when compared to untransfected Jurkat cells or Jurkat cell lines transfected with the parent control plasmid. Sublines derived from the high-level Tat transfectants with reduced Tat activity showed normalization of PHA/PMA-induced IL-2 expression. Northern analysis showed diminished expression of IL-2 and IL-2R mRNA in the stimulated Tat transfectants. Inhibition of IL-2 and IL-2R expression by the HIV-1 Tat protein may contribute to the immune suppression that characterizes HIV-1 infection.


Inhibition of in vitro T cell activation by corneal endothelial cells.

Cells and tissues of the anterior uvea and aqueous humor express activities which inhibit immune responses. These activities include soluble factors such as TGF-beta and uncharacterized cell surface interactions. Relatively little is known regarding the immunologic activities of corneal endothelium, despite its potentially important role in contributing to the immune privilege of the anterior chamber and the high success rate of corneal transplantation. In this report, in vitro studies of cultured rat corneal endothelial (CE) cells were done using S-antigen-specific LEW rat T cell lines, or S-antigen-specific T cell hybridomas, to examine the immunologic capabilities of CE cells. Monolayers of LEW rat CE cells were unable to present antigen or a mitogen, Con A, to T cell lines or hybridomas as assessed by the lack of a proliferative response or IL-2 secretion. Furthermore, the CE cells exerted a potent inhibitory effect when added to in vitro proliferation assays of T cell lines stimulated with antigen or Con A. When T cells were preactivated on conventional antigen presenting cells and then transferred to wells containing CE cells, their proliferation was not inhibited. Although CE cells inhibited activation of T cell lines and hybridomas, they did not inhibit the growth of T cell hybridomas or CTLL cells, nor did the CE cells adversely affect the viability of resting T cells cultured on CE monolayers. The inhibitory effect was reversible as preincubation of T cells on CE cells for up to 6 days followed by washes restored T cell responsiveness when assayed on splenocytes. The inability to stimulate proliferative responses was not affected by preincubation of the CE cells with lymphokines which increase MHC antigen expression. The inhibition observed in these assays was not MHC-restricted as CE cells from both LEW and BN rats were equally inhibitory. CE cells from rabbits and cats were also potent inhibitors of T cell activation, suggesting that the mechanism is evolutionarily conserved. The mechanism of inhibition of CE cells is unknown at this time.


Sensitization of rat alveolar macrophages to enhanced TNF-alpha release by in vivo treatment with dexamethasone.

Treatment of rats with dexamethasone rapidly induced a marked weight loss which occurred within 3 days and persisted for several weeks. The cachectic state was paralleled by increased serum levels of triglycerides, albumin, and protein and a strong reduction of blood mononuclear leukocytes. In lung sections, an increased number of mononuclear giant cells was found but no bacteria, fungi, or Pneumocystis carinii organisms. Quite strikingly, alveolar macrophages from dexamethasone-treated rats, but not from control animals, were highly sensitive to LPS and released large amounts of TNF-alpha ex vivo. Also under in vivo conditions, high TNF-alpha serum concentrations were found in dexamethasone-treated but not control rats when examined 1 1/2 hr after an intravenous LPS injection. These data suggest that the glucocorticoid-induced cachexia of rats may be linked, at least in part, to readily inducible TNF-alpha release from primed macrophages.


Specific T-cell factor production and lymphocytes in the direct surroundings of a subcutaneous allogeneic tumor.

Antigen-specific T-cell factors (TCF) play a role in the initiation of cellular immune responses. In allogeneic mouse-tumor models lymphocytes from the direct tumor surroundings of both euthymic and nude mice produce TCF. These lymphocytes produce TCF when collected already 1 day after subcutaneous (sc) injection of tumor cells. In contrast to euthymic mice, draining lymph nodes and spleen of nude mice did not contain TCF-producing lymphocytes at any stage after sc tumor cell injection. In sensitized euthymic mice TCF production by lymphocytes is significantly higher in the direct tumor surroundings than in draining lymph nodes or spleen. At 2 and 5 days after tumor cell injection, the mononuclear cell infiltrate of the tissue surrounding the tumor in euthymic mice showed low expression of Thy 1, CD3, TCR alpha beta, TCR gamma delta, CD4, CD8, and asialo GM1, whereas several lymphocytes and mast cells were positive for monoclonal antibody (mAb) 14-30 (directed against TCF). In both euthymic and nude mice, sc injected tumor cells showed apoptosis. In conclusion, the direct tumor surroundings are the first (and, for nude mice, the only) site of TCF production, sc injection of tumor cells attracts mAb 14-30-positive lymphocytes and renders mast cells positive for mAb 14-30.


Effect of immunosuppressive therapy on cytolytic activity of immunodeficient mice: implications for xenogeneic transplantation.

Despite major deficits in their immune system, SCID, Nude, and NIH III mice reject allo- and xenografts, particularly leukemic cell lines, albeit less readily than immunologically intact mice. Since variation among these immunodeficient mouse strains in rejection of a human lymphoid cell line (CCRF-CEM) parallels splenic non-MHC-mediated cytolytic activity, non-MHC-restricted cytolytic activity may be responsible for retained resistance to leukemic cell transplantation. SCID mice that had the least cytolytic activity accepted 100% of their grafts. The converse was true for NIH III mice that showed the greatest cytolytic activity and were relatively resistant to CEM cell engraftment. Different approaches to ablate NK activity and thus enhance engraftment led to variable results for each strain. A single dose (500 micrograms) of anti-asialoGM1 (AsGM1) markedly reduced NK activity in SCID and NIH III mice by 60 and 40%, respectively. A moderate 20% decrease was seen in Nude mice at this dose. In contrast, gamma irradiation suppressed NK activity by greater than 80% of baseline levels in all three strains. Of importance, total cytolytic activity in immunosuppressed Nude and NIH III mice, although significantly depressed compared to untreated mice of the same strain, still remained higher than that seen in nonimmunosuppressed SCID mice. Enhanced engraftment and systemic dissemination of CEM cells in immunosuppressed mice correlated directly with decreased total splenic cytolytic activity in all three strains. These results have implications for the use of immunodeficient models for transplantation, tumor immunobiology, and engraftment of a human immune system.


A study of autologous anti-idiotypic antibody-forming cells in mice of different ages and genetic backgrounds.

Antibody response to phosphorylcholine, an immunodominant epitope of Streptococcus pneumoniae R36a (Pn), is characterized by a public idiotype, T15, that is expressed on a large proportion of antibody molecules produced by all mouse inbred strains. The ability of the immune system to produce an autologous antibody to T15 upon immunization with Pn vaccine was investigated using a modified ELISA plaque assay for detection of single antibody-forming cells (AFC). The limit of ELISA assay for detection of specific anti-T15 AFC is approximately 300 cells/spleen. However, our studies failed to detect any autologous anti-T15 AFC in the course of the primary antibody response to Pn vaccine in young/adult (2-4 months) BALB/c and C57BL/6 mice. Aged mice (20-22 months) also failed to develop any specific auto-anti-T15 AFC upon the primary Pn immunization, despite the fact that the anti-Pn response in these animals changes both quantitatively and qualitatively. In order to generate specific anti-T15 AFC, BALB/c mice had to be immunized repeatedly with Pn vaccine (four weekly injections) or immunized directly with T15 protein in CFA. Different results were obtained with D1.LP mice that are low responders to Pn and express lower levels of T15 Id as compared to BALB/c. Young D1.LP mice produced high numbers of auto-anti-T15 AFC of both IgM and IgG isotypes following a single immunization with Pn vaccine. The kinetics of auto-anti-T15 response in D1.LP mice was similar to that of the antigen-specific response. These results demonstrate that the ability of the immune network to produce autologous antibody to a shared Id depends on the genetic makeup of the host, and that this response may be regulated by the level of Id expression.


Tumor cytostasis mediated by LPS- or PSK-activated human plastic-adherent peripheral blood mononuclear cells.

We investigated the mechanism of cytostasis mediated by activated human plastic-adherent peripheral blood mononuclear cells (PBMC) in two cell lines, L.P3 cells (TNF alpha sensitive) and A375 cells (TNF alpha insensitive), using two biological response modifiers, lipopolysaccharide (LPS) and a protein-bound polysaccharide extracted from a fungus, PSK. In L.P3/LPS, L.P3/PSK, and A375/LPS cultures, the cytostatic effects were significantly reversed by anti-TNF alpha antibody, while in the A375/PSK culture they were not. In concordance with this, LPS was a good inducer of TNF alpha, but PSK was not. In A375/PSK culture, PSK-activated cells arrested A375 cells at the boundary between G1 and S, presumably through inhibition of polyamine synthesis. This growth inhibition may be mediated by an unknown soluble factor which is different from TNF alpha, IL-1, IL-6, and TGF beta.


Characterization of T cell proliferative responses induced by anti-Qa-2 monoclonal antibodies.

The MHC class I Qa-2 Ag are attached to the cell surface by a glycanphosphatidylinositol (GPI) anchor. Crosslinking of Qa-2 and several other cell surface Ag attached by the GPI linkage has been shown to lead to cell activation. We have developed 10 new anti-Qa-2 mAb and characterized their capacity to induce proliferation of spleen cells. In the absence of anti-Ig-mediated crosslinking, none of the mAbs alone could induce activation. However, mAb 23.1 which reacts with the alpha 3 domain of Qa-2, when combined with most of the other mAbs (alpha 1, alpha 2 domain reactive), activated cells in the absence of anti-Ig crosslinking. The mAb pair 23.1 plus 24.16 was the most proficient and induced proliferation in the absence of any exogenous second signals. Responses were greatly enhanced and equivalent to those seen with anti-CD3 by the addition of phorbol myristate acetate (PMA). Ionomycin, rIL-2, or rIL-4 also potentiated anti-Qa-2 responses but less efficiently than PMA. Significant strain variation in the magnitude Qa-2-mediated proliferative responses was observed correlating with the levels of Qa-2 expressed on the cell surface. Crosslinking of Qa-2 molecules by the mAb combinations was required because monovalent Fab fragments failed to activate cells. F(ab')2 fragments of mAb 23.1 plus 24.16 induced vigorous proliferation indicating that accessory cell presentation of the mAb via Fc receptors was not required. Immobilized (plate bound) anti-Qa-2 mAb induced proliferation suggesting that the Qa-2 pathway may be distinct from that of other GPI molecules such as Thy-1 and Ly-6. Populations enriched for T cells (approximately 95%) responded as well as whole spleen cells, whereas B lymphocytes failed to proliferate to anti-Qa-2. Both CD4+ and CD8+ cells were activated following crosslinking of Qa-2. Finally, T cell activation mediated by Qa-2 induced elevation of [Ca2+]i, IL-2R expression, and the release of IL-2. These data demonstrate that crosslinking of Qa-2 on T lymphocytes represents a potent pathway for inducing cell activation.


Suppression of chronic antigen-induced arthritis in rats by a monoclonal antibody against the T cell receptor alpha beta.

We have investigated a role for T cells in chronic antigen-induced arthritis in rats employing a monoclonal antibody (R73 mAb) against the T cell receptor alpha beta. Treatment with R73 mAb from the time of intra-articular antigenic challenge blocked completely the induction of chronic, but not acute ovalbumin-induced arthritis in sensitized rats. Histologically, treatment-controlled arthritic rats exhibited marked hyperplasia of synovial membrane with pronounced infiltration of inflammatory cells including alpha beta + T cells in the chronic phase of arthritis. In contrast, R73 mAb-treated rats had almost normal joint histology. Treatment with R73 mAb after onset of arthritis was also effective in suppressing the progression of chronic antigen-induced joint inflammation. The preventive and suppressive effects of the mAb on chronic antigen-induced arthritis were associated with marked depletion of alpha beta + T cells in peripheral blood. The DTH but not the humoral response to ovalbumin in sensitized rats was suppressed significantly by R73 mAb. Thus, alpha beta + T cells appear to have a central role in both induction and progression of chronic antigen-induced arthritis.


Comparison of the capacity of murine and human class I MHC molecules to stimulate T cell activation.

The mechanism underlying the apparent differences in the capacity of murine and human class I MHC molecules to function as signal transducing structures in T cells was examined. Cross-linking murine class I MHC molecules on splenic T cells did not stimulate an increase in intracellular calcium ([Ca2+]i) and failed to induce proliferation in the presence of IL-2 or PMA. In contrast, modest proliferation was induced by cross-linking class I MHC molecules on murine peripheral blood T cells or human class I MHC molecules on murine transgenic spleen cells, but only when costimulated with PMA. Moreover, cross-linking murine class I MHC molecules or the human HLA-B27 molecule on T cell lines generated from transgenic murine splenic T cells stimulated only modest proliferation in the presence of PMA, but not IL-2. On the other hand, cross-linking murine class I MHC molecules expressed by the human T cell leukemic line, Jurkat, transfected with genes for these molecules, generated a prompt increase in [Ca2+]i, and stimulated IL-2 production in the presence of PMA. The results demonstrate that both murine and human class I MHC molecules have the capacity to function as signal transducing structures, but that murine T cells are much less responsive to this signal.


Priming of peripheral lymph node B cells with TNP-Ficoll: role of lymphokines in B cell differentiation.

We have previously shown that peripheral lymph node (PLN) B lymphocytes of adult DBA/2J mice failed to make an antibody response to type 2 antigen TNP-Ficoll, but exhibited a good antibody response to type 1 antigen TNP-Brucella abortus. In the present study we wanted to find out whether the unresponsiveness of PLN B cells to TNP-Ficoll is due to defects in the early activation and proliferation stage or in the final differentiation stage of B cells. Therefore, we have used a two-step protocol of in vivo immunization of mice with TNP-Ficoll and the subsequent in vitro challenge with TNP-Brucella abortus and studied the anti-TNP plaque-forming cell (PFC) responses. The results indicate a three- to sixfold increase of PFC responses in PLN cell cultures derived from TNP-Ficoll-primed animals compared to saline control mice. This increased antibody response was TNP-specific as 93% of the PFC's were inhibited by TNP-lysine. Limiting dilution experiments confirm that the increase in anti-TNP PFC response from the TNP-Ficoll-primed animals was indeed due to an increase in TNP-specific precursor B cells. Further, the addition of rIL-5 or rIL-6 induced anti-TNP PFC in the TNP-Ficoll-primed and in control PLN cell cultures in the presence of antigen. However, in primed PLN cells lymphokines alone were sufficient to restore anti-TNP PFC response. In conclusion, our results show that in PLN, the TNP-Ficoll can induce proliferation of hapten-specific B cells but not final differentiation. These primed PLN B cells mature into antibody-secreting cells upon stimulation with TNP-BA or lymphokines.


Transforming growth factor-beta inhibits the production of IgG, IgM, and IgA in human lymphocyte cultures.

Transforming growth factor beta (TGF beta) has potent immunoregulatory effects acting on both T and B cells. It strongly inhibits secretion of IgG and IgM in human and murine B cell cultures, but has been shown to have an enhancing effect on IgA production in the mouse. We have studied the effect of TGF beta on the production of IgA in human lymphocyte cultures. The addition of TGF beta to pokeweed-stimulated peripheral blood lymphocytes resulted in a suppression of IgA production of both subclasses, similar in magnitude to the suppression of IgG and IgM production. Membrane IgA expression was not increased by culturing tonsillar lymphocytes with TGF beta. In conclusion, we find no evidence for a selective enhancing effect of TGF beta on IgA synthesis in humans, in contrast to the findings reported in mice.


Induction of syngeneic cytotoxic T lymphocytes against a B cell tumor. II. Characterization of anti-idiotypic CTL lines and clones.

In an earlier communication we showed that idiotypic immunoglobulin (Id+ Ig) of a B cell hybrid, 2C3, can induce cytotoxic T lymphocytes (CTL) in the spleens of mice that are hyperimmunized with the irradiated tumor cells. To understand the extent of heterogeneity in the splenic CTL population, stable anti-idiotypic CTL lines and clones were established from 2C3-primed splenocytes. One representative CTL line A102 which exhibited the phenotype of CD3+, CD4-, and CD8+, has been maintained in long-term culture for more than 18 months. Cytotoxic specificity of A102 was determined by cold target inhibition assay using a panel of syngeneic and allogeneic B cell tumors. The CTL line A102 was highly cytotoxic to 2C3, only weakly to other syngeneic tumors, but not at all to allogeneic B cell tumor CH12. Furthermore, CTL-mediated cytolysis was significantly abrogated by blocking 2C3 cells with anti-idiotypic monoclonal and polyclonal antibodies. These results clearly show that 2C3 Id represents the immunodominant epitope(s) recognized by the CTL line A102. To isolate a highly Id-specific effector population, A102 was repeatedly subcloned by limiting dilution. One such clone 102.F5 exhibited considerable specificity toward Id+ 2C3 while another clone 102.E10 showed no such specificity in a competitive cytotoxicity assay. This was further confirmed by the inhibition studies with anti-Id mAb. Thus, hyperimmunization with irradiated 2C3 cells evokes a spectrum of anti-2C3 cytotoxic effector cells, of which a major population is reactive to the idiotypic determinants associated with 2C3 Ig.


Induction of syngeneic cytotoxic T lymphocytes against a B cell tumor. III. MHC class I-restricted CTL recognizes the processed form(s) of idiotype.

The purpose of this work was to determine the molecular nature of the idiotypic Ig of a B cell tumor, 2C3, involved in the induction of anti-idiotypic cytotoxic T-lymphocytes (CTL). We previously reported that hyperimmunization of mice with irradiated 2C3 cells provides effective tumor protection by inducing MHC class I-restricted CTL. Due to the enormous heterogeneity of the splenic CTL further study could not be undertaken on the idiotype (Id)-CTL interaction. Subsequently an anti-idiotypic CTL line, A102, and a highly Id-specific CTL clone, 102.F5, have been developed. In the present investigation we report that the processed forms of idiotypic determinants are responsible for induction and activation of these specific effector CTL. Inhibition studies using anti-TcR and anti-MHC class I mAbs showed that the TcR-CD3 complex of the anti-idiotypic CTL recognized 2C3 Id in the context of MHC class I antigens. The cytotoxicity of these CTL could not be inhibited with affinity-purified 2C3 Ig used as such or after pulsing with splenic antigen-presenting cells (APC). Furthermore, using brefeldin A (BFA) and chloroquine (CLQ), which are specific inhibitors of cytosolic and endosomal antigen processing pathways, respectively, it has also been observed that exposure of 2C3 to BFA but not CLQ prevents its cytolysis by both anti-idiotypic CTL line and clone. These results clearly indicate that endogenously produced idiotypic determinants of 2C3 Ig are processed in pre-Golgi vesicle, possibly in the ER, along with MHC class I antigens and then are transported to the membrane. Treatment of 2C3 with BFA, however, did not exert any effect on the expression of membrane-associated Ig of 2C3 cells. Therefore, it is the processed form rather than the bona fide receptor Ig on the cell surface that is recognized by the Id-specific CTL.


Gene transfer into mammalian somatic cells in vivo.

Direct gene transfer into mammalian somatic tissues in vivo is a developing technology with potential application for human gene therapy. During the past 2 years, extensive progress and numerous breakthroughs have been made in this area of research. Genetically engineered retroviral vectors have been used successfully to infect live animals, effecting foreign gene expression in liver, blood vessels, and mammary tissues. Recombinant adenovirus and herpes simplex virus vectors have been utilized effectively for in vivo gene transfer into lung and brain tissues, respectively. Direct injection or particle bombardment of DNA has been demonstrated to provide a physical means for in situ gene transfer, while carrier-mediated DNA delivery techniques have been extended to target specific organs for gene expression. These technological developments in conjunction with the initiation of the NIH human gene therapy trials have marked a milestone in developing new medical treatments for various genetic diseases and cancer. Various in vivo gene transfer techniques should also provide new tools for basic research in molecular and developmental genetics.


Roles of immobilized glycosylated proteins and lipoprotein(a) in adhesivity of endothelial cells: possible implications for atherogenesis.

The adherence of peripheral blood monocytes to adult bovine endothelial cells grown to confluence in the microplates was studied. The microplates were coated with different proteins or used without special treatment. It was shown that endothelial cells seeded on immobilized glycosylated proteins (serum albumin or skin gelatin) adhered more monocytes than the cells grown on non-modified proteins. Endothelial cells grown in lipoprotein(a) coated wells bound more monocytes than the cells grown in non-treated microplates or in wells coated with low density lipoproteins (LDL). The effect of lipoprotein(a) coating could not be reproduced by treating the plates with plasminogen (as a homolog of apo(a)) or with a mixture of LDL and plasminogen. These results indicate that the composition of extracellular substrata has a profound effect on adhesive properties of cultured endothelial cells. The implications of these findings for atherogenesis and for the general aspects of regulation of cell adhesion are discussed.


An acid extract from dissociation medium of sea urchin embryos, induces mesenchyme differentiation.

When material extracted by 1 M acetic acid from the dissociation medium of sea urchin embryos is added at low concentrations to isolated primary mesenchyme cells, it induces skeletogenesis. The same material added to dissociated blastula cells, or to embryos at the blastula stage, stimulates skeleton formation and pigment cell differentiation. On dissociated cells, it also increases cell reaggregation, thymidine incorporation and survival. On embryos, it induces exogastrulation and appearance of extraembryonic pigment cells. The activity of the extract is resistant to raised temperatures and partially to tryptic digestion but is abolished by trypsin treatment followed by heating. The active fraction does not readily filter through Amicon XM-50 and is retarded by column chromatography on Bio-Gel P-60.


The inhibitory effect of anthranilate derivatives on HCO3-/Cl- exchange in red blood cells of human, pigeon and trout.

All ten of anthranilate derivatives were found to be inhibitors of HCO3-/Cl- exchange in human, trout and pigeon red blood cells. The individual l50 values covered a range of 2-3 orders of magnitude for human, trout and pigeon RBCs. The results obtained confirm that the anion transporting region of the band 3 protein in human, trout and pigeon red blood cells are highly preserved even though the RBCs belong to animals living in entirely different habitats. However, in spite of the general similarities, significant differences between human trout and pigeon red blood cells could be observed. They could be attributed to differences of the hydrophobic character of the various probes.


Timing of mitotic chromosome loss caused by the ncd mutation of Drosophila melanogaster.

We studied the timing of mitotic loss of maternally and paternally derived chromosomes among the progeny of Drosophila melanogaster females homozygous for an amorphic mutation in ncd, a gene encoding a kinesin-like protein. In order to determine the division at which chromosome loss occurs, we estimated the fraction of XO nuclei resulting from X chromosome loss by scoring the phenotype of 47 adult cuticular landmarks in 160 XX-XO mosaics (gynandromorphs) derived from maternal X chromosome loss, and 33 gynandromorphs derived from paternal X chromosome loss. The results show that while most of the mitotic loss of maternally derived chromosomes occurs at the first cleavage division, the mitotic loss of paternally derived chromosomes occurs only at the second and later divisions. This means that paternally derived chromosomes are immune from the effects of ncd prior to karyogamy, which occurs after the first cleavage division. We discuss the implications of these results for the function of the ncd gene product and for other kinesin-like proteins in Drosophila.


Distribution of detyrosinated microtubules in motile NRK fibroblasts is rapidly altered upon cell-cell contact: implications for contact inhibition of locomotion.

Fibroblasts migrating into an experimental wound contain an extensive array of detyrosinated microtubules (Glu MTs) oriented in the direction of migration, whereas nonmotile cells in the interior of a monolayer contain Glu MTs that are primarily coiled around the nucleus. To determine the role of cell-cell contact in the formation of these distinct arrays of Glu MTs, we studied the distribution of Glu MTs by immunofluorescence in NRK fibroblasts that had been fixed at different intervals after they had established contact with other cells. Time-lapse video recordings were made of the contacting cells to provide a record of cellular behavior. In motile cells that became completely surrounded by virtue of contact with other cells, Glu MTs were found mostly coiled around the nucleus. The proportion of cells whose Glu MTs extended to the original leading edge decreased dramatically after the cells had been surrounded for 10 min or more. At earlier times, when the contact was confined to a portion of the cell margin, Glu MTs were absent from the area behind the contact site, yet were still oriented toward the noncontacting and ruffling margins. The contact-induced alteration of Glu MTs was not due to the cessation of forward locomotion of cells per se, since immobilization of cells with cytochalasin D did not cause a dramatic change in Glu MTs. That cell-cell contact also specifies the type of Glu MTs formed in cells was shown by experiments in which MTs were regrown following complete depolymerization with nocodazole. The remodeling of Glu MTs during cell-cell contact may be involved in cellular repolarization during contact inhibition of locomotion and will be a useful marker for further dissecting the molecular events of contact inhibition of motility.


Evolution of the flagellar waveform of ram spermatozoa in relation to the degree of epididymal maturation.

Motility and flagellar movement of ram spermatozoa along the epididymis were analysed in vitro. From the caput to the cauda of the epididymis, the percentage of motile and progressive spermatozoa increases. No flagellar bending was observed in spermatozoa from the testis or the epididymal anterior caput. When spermatozoa reached the distal caput of the epididymis, a static curvature, associated with an initiation of the flagellar beating, appeared on the flagella. This curvature normally disappeared during epididymal transit. Its disappearance was associated with an increase in the flagellar beat efficiency. Our results suggest that the initiation of motility is related to two mechanisms involving: (1) the presence of a transient static curvature, and (2) the establishment of a symmetric regular beating of the flagellum.


Image blurring by thermal diffusion in the observation of hydrated biomolecules with soft X-ray microscopy.

A simple model is presented for the estimation of image blurring in X-ray microscopy of biological specimens in a hydrated environment. The model is essentially based on thermal diffusion of an object to be imaged. The degree of image blurring by diffusion depends on the following situations of the object. The object is free from, is tightly fixed to, or is partially connected to the surrounding structures. The proper imaging time required to achieve a given resolution in X-ray microscopy of biological structures was estimated with the present method. The results suggest that imaging time shorter than 3 msec (free) to 1.4 sec (tightly fixed) is required for the observation of a cell (30 microns in diameter) at the resolution of 100 nm. The model is also applicable to a fragmented object caused by imaging X-rays.


Novel monoclonal antibody mAb G3A5 recognizes 138-kDa glycoprotein localized on the Golgi membrane.

Partially purified Golgi membranes of HeLa cells were used as antigen to produce a novel monoclonal antibody (mAb G3A5). The mAb G3A5 specifically labeled Golgi apparatus of human and monkey cultured cells as ascertained by indirect immunofluorescence but did not stain those of bovine or mouse cells. Treatment with nocodazole and brefeldin A (BFA) induced fragmentation and redistribution of the staining. Western immunoblot analysis showed that mAb G3A5 was directed against a single polypeptide with an apparent molecular mass of 138-kDa (p138 antigen). The p138 antigen is an integral membrane protein of the Golgi apparatus, as assessed by several assays: protease protection, salt wash and flotation in sucrose density gradient centrifugation. The p138 antigen was purified using immunoaffinity chromatography. The apparent molecular mass of the p138 antigen decreased by 2 to 4 kDa after treatment with the peptide: N-glycosidase F, while digestion with ENDO F or Neuraminidase did not have this effect. Thus, p138 antigen is a glycoprotein containing asparagine-linked carbohydrates.


A cell line derived from sphingomyelinosis mouse shows alterations in intracellular cholesterol metabolism similar to those in type C Niemann-Pick disease.

Cell lines derived from the sphingomyelinosis (gene symbol, spm) mouse were established from homozygous (spm/spm) and heterozygous (spm/+) embryos according to a rigid 3T3 transfer schedule. The SPM-3T3 cells derived from a homozygous embryo showed extensive accumulation of intracellular cholesterol, attenuated esterification of exogenously added cholesterol and increased de novo cholesterol synthesis, when compared to SPMH-3T3 cells derived from a heterozygous embryo. The phenotypic abnormalities were very similar to those observed in fibroblasts from patients with Niemann-Pick disease type C (NP-C), in which a defect in the intracellular transport of unesterified cholesterol is suggested. The genetic defect in SPM-3T3 cells should be closely related to that in NP-C. The SPM-3T3 cell line is useful for biochemical and genetic studies on the regulation of intracellular cholesterol metabolism.


[Use of the immunoblotting method in serodiagnosis of M. kansasii mycobacteriosis].

The indirect enzyme test ELISA with soluble complex antigen of M. kansasii, used for assessment of the titre of IgG serum antibodies in a group of patients suffering from mycobacteriosis M. kansasii and in a control group of patients suffering from tuberculosis did not reveal statistically significant differences between serological responses of these two groups. On the other hand, the immunoblot analysis revealed in sera selected at random from these two groups differences at the level of subprotein units against the complex antigen of M. kansasii. In a group of 15 sera of patients with M. kansasii the immunodominant area was 42-45 kD protein subunits, in the control group of 10 sera with M. tuberculosis the area was 35-39 kD. The western blot technique is more perspective for improvement of the serum diagnosis of M. kansasii, in particular where a specific antigen is not available.


[A methods of processing and microbiological study of sputum].

In the submitted paper the author discusses quantitative and qualitative methods of examination of sputum, using different reagents for blending and lysis of sputum. The micromodification of Dixon-Miller's method is most effective from the economic aspect and as regards quality of the examination. The dilution method makes it possible, using selective media, to complete the whole examination in a shorter time without using special equipment for cultivation and assessment of sensitivity to antibiotics in ordinary microbiological field laboratories.


[Detection of mycobacteria in pathologic material using the radiometric method].

For concurrent detection of mycobacteria by the radiometric method BACTEC and the standard cultivation technique 96 samples of pathological material from patients hospitalized with suspected or confirmed tuberculosis was used. The total number of positive samples was 30, using the BACTEC method only, four samples and using the cultivation method only, 10 samples. The assessed difference was not statistically significant. The mean period of positivity with the BACTEC method was 20.4 days, in cultivations 28.8 days. The higher yield of the cultivation method could be caused by various factors. For the BACTEC system inocula of 0.5 ml were used and one cultivation bottle, for the cultivation inocula 1.0 ml and four test tubes for one sample were used. From five samples, positive only on cultivation, only sporadic colonies were obtained after 42-63 days cultivation and in all instances only on one of four media.


[The present epidemiologic status and prognosis in measles].

During 1990 and partly also 1991, after a practically 10-year zero incidence of measles on the territory of the Czech Republic a measles epidemic broke out. The most severely affected group were adolescents aged 15-19 years who were immunized only by a single dose of vaccine. Conversely the lowest morbidity was recorded in 2-9-year-old children who were immunized already with two doses. The cause of the epidemic was primary failure of the vaccine leading to a low level of collective immunity in some population age brackets. The authors discuss and explain the strategy of immunization against measles involving two doses of vaccine after a 6-10-month interval. In the conclusion possibilities to achieve permanent elimination of measles on our territory are discussed.


[Isolation of a rare serovar of Plesiomonas shigelloides].

The authors describe the isolation of a rare serovar Plesiomonas shigelloides from a patient with diarrhoea. The isolated strain belongs according to a combined Japanese-Czechoslovak antigenic scheme to serovar 024 H"o". The interesting finding draws attention to the possibility of routine departments to contribute to investigations of the incidence, importance and antigenic structure of this bacterial species.


[Knowledge of high school students about HIV and AIDS].

In 475 students attending the last school year in four high schools in the town of Povazská Bystrica, Slovakia a questionnaire survey of students' knowledge of HIV infection and AIDS was carried out. 76.33% of all answers to questions in the questionnaire were correct. We found no significant differences between the percentage of correct answers among schools, among boys and girls and among students living in the country and in town. 56% of the students fought that there is enough information about HIV infection, AIDS and about prevention of that infection. Nevertheless the percentage of correct answers to some questions in the questionnaire do not correspond to this statement.


[Bacterial contamination of arthropods in a health facility].

At 55 sites of a health institution in July and September 1990 a total of 161 specimens of arthropods were detected, 30 outdoors and 131 on the premises of the health institution. On their bodies 116 bacterial strains were isolated, mostly Gram-negative rods (more than 85%), in particular spp. Enterobacter, Acinetobacter, Klebsiella, Citrobacter and Pseudomonas. Gram-positive cocci accounted for cca 12%, in particular strains of S. haemolyticus and S. hominis. The greatest number of strains was detected on bodies of cockroaches, flies, Chironomus and Tenebrio. In about one third of strains the diffuse disk test revealed resistance to more than three antibiotics. The investigation was supplemented by microbiological examination of strains from a hospital environment (45 smears) and strains from biological material (82 specimens), from patients with nosocomial infections.


[Helminthologic examination of swimming pool water in Czechoslovakia].

The author evaluated conditions necessary for the development of the most common intestinal helminths in swimming pools and emphasizes provisions which prevent their development. The author recommends helminthological examinations only in indicated cases.


[Examination of ticks for the presence of Borrelia sp. in Kosice and the surrounding region. Preliminary results].

The authors present preliminary data on the infestation of ticks Ixodes ricinus with Borrelia in the town of Kosice and surroundings. The authors found a mean 4.7% positivity which means that more systematic attention should be paid to the problem.


[Detection of Chlamydia trachomatis in clinical material 1989-1990].

Examination of 276 smears and scrapings from the uterine cervix of women before delivery and during the puerperium revealed in 28.0% positive findings. Perinatal infection was confirmed by the presence of Chlamydia trachomatis in 35.5% neonates with conjunctivitis and 27.3% positive smears from the nasopharynx. The authors used the direct immunofluorescence method. For immunofluorescent staining they used Chlamyset of Orion Co., Finland.


[Antimicrobial activity of selected aqua-carboxyl-cupric complexes].

The activity of compounds of different structural types of aqua-complexes of the composition Cu(R-COO)2.nH2O-methoxybenzoatocupric complexes, R = 2-, 3- and 4-methoxyphenyl (n = 1, 1 and 3); aryloxyacetatocupric complexes, R = phenoxymethyl (n = 3), 2-, 3- and 4-chlorophenoxymethyl (n = 4, 2 and 2) and 1-naphthoxymethyl (n = 4), and furthermore isomeric furanecarboxylato-(R = 2- or 3-furyl, n = 3, or 1) and thiophencarboxylatocupric complexes (R = 2- or 3-thienyl, n = 1 and 1), was examined by the methods of the 1st screening on selected anthropo- and phytopathogenic microorganisms. The effects of all aqua-complexes (suspension dosage form) on the representatives of bacteria and yeasts are minimal. On the other hand, the effect of these substances on the causative agents of dermatomycoses (Trichophyton terrestre, Microsporum gypseum) in the case of methoxybenzoato- and furoatocupric complexes achieves a MIC value of 500 micrograms/cm3 and lower. The activity against phytopathogenic fungi (both in vitro and in vivo experiments) is generally relatively low at 0.05% concentration of active ingredients (dispersible powders). At the same time the antimicrobial activity of the pertinent free carboxylic acids was investigated in relation to the cupric salts being formed. They are able to form the required pharmacoactive form prevalently as late as they are in the form of aqua-carboxylatocupric complexes.


[Does the spermatozoa survival test correlate with fertilization of oocytes in vitro?].

The spermatozoa survival test (SST)--which is used in our laboratory is considered to be a relatively simple test with a large probability determining the ability of spermatozoa to fertilize oocytes in vitro. The SST reveals the percentage of motile spermatozoa after 20hr. cultivation with an oocyte. 221 sperm samples used for oocyte fertilization were analyzed and divided into four groups: normospermia (123 samples), oligospermia (19), asthenospermia (45) and oligoasthenospermia (34). The results show a significant reduction of the fertilization rate (< 35%) below the range of 40% SST in all groups. Above the value of 40% motile spermatozoa after 20hr. cultivation is the fertilization rate in all groups higher (> or = 50%). The authors recommend the SST as a part of the screening examination before the couple is admitted to an assisted reproduction programme.


[Results of surgical treatment of ovarian dysfunction].

The authors operated 155 women where ovarian dysfunction was the cause of infertility. The first group was formed by 125 women with the Stein-Leventhal syndrome, the second group was formed by 30 women with dysgenesis of the gonads, karyotype, 46,XX. In the first group 97 of the operated women (77.6%) became pregnant. In the latter group after mere resection of the gonads 45 women (36%) became pregnant. After combined therapy (surgery and hormonal therapy) 52 infertile women (41.6%) became pregnant. Twenty-eight patients (22.4%) did not become pregnant. In the second group, i.e. in the group of gonadal dysgenesis 7 of 16 women (44%) became pregnant but only in the group of sclerocystic dysgenetic gonads. None of the women with streak or hypoplastic gonads became pregnant. In gonadal dysgenesis it is important to assess the quality of the follicular apparatus. For successful surgery of the ovary it is necessary to preserve a maximum of functional tissue and to use a careful surgical technique.


[Treatment of vesicovaginal fistulas].

The authors evaluated in a retrospective analysis their experience with treatment of vesicovaginal fistulae in 36 patients. Most frequently--almost in 90%--the fistulae developed after a gynaecological operation, in particular surgery on account of benign, but also malignant disease. The authors used for surgical correction of the fistulae a transvaginal approach in 28 (77.8%) patients, a transvesical approach in 5 (13.9%) patients and a transvesical and transperitoneal approach in two (5.5%) patients. One fistula receded spontaneously during drainage of the urinary bladder. Primary correction of the fistula was successful in 34 (94.4%) patients. In two patients further surgical treatment was necessary. With regard to their favourable experience the authors recommend a transvaginal approach which is in their opinion suited for the majority of non- complicated vesicovaginal fistulae. In corrections of extensive fistulae after radiotherapy a combined transvesical and transperitoneal approach with fixation of the omentum in the space between the vagina and urinary bladder proved useful.


[Diagnosis of malignant ovarian tumors using color Doppler sonography].

In a prospective investigation the authors followed the circulation in 32 patients with ovarian tumours before operation. They examined the RI using colour Doppler sonography. Based on histological examination the authors divided the patients into groups with malignant and benign tumours resp. The peripheral resistance and RI in the group of patients with malignant tumours was 0.35 +/- 0.03, i.e. significantly lower than in the group with benign tumours, 0.65 +/- 0.08 (p < 0.001). In none of the patients with malignant tumours the RI was higher than 0.50. The authors recommend this method as a screening examination for early detection of malignant ovarian tumours.


[Detection of intrauterine fetal growth retardation using ultrasound fetometry].

Based on ultrasonic foetometry at an interval of 0-59 days before delivery the authors assessed in 93 women with risk pregnancies hospitalized during the 31st-43rd week of gestation the index of the femur length/abdominal circumference independent on gestation age, and they estimated the foetal weight by means of three logarithmic formulae comprising the biparietal diameter, femur/length and abdominal circumference. For detection of foetal hypotrophy, the estimated weight of the foetus in particular according to the formula of Hadlock et al. (sensitivity 58.3%, specificity 94.22%) was significantly more accurate than the index of femur length/abdominal circumference > or = 23.0 (sensitivity 45.8%, specificity 91.3%).


[Prophylactic administration of Claforan in patients with premature loss of amniotic fluid].

The authors compared two groups of pregnant women with early loss of amniotic fluid. In the first group Claforan was administered, 3 x 1 g by the i.m. route, and in the second group Ampicillin, 3 x 1 g by the i.m. route. The data were recorded in a special protocol and were divided into three groups: 1. patient before delivery, 2. patient during puerperium, 3. neonate. From the results two conclusions were drawn: 1. The authors did not detect any reasons why antibiotics should not be administered prophylactically in case of premature loss of amniotic fluid. 2. The main differences between the compared groups were found in neonates where after administration of Claforan there was a substantially lower incidence of positive bacterial cultures than after Ampicillin and there was also a lower incidence of RDS II and adnatal infections.


[Does transabdominal amniocentesis in the 2d trimester affect fetoplacental circulation?].

In a prospective study the authors focused attention on Doppler sonographic examination of the foetoplacental circulation and whether it is influenced by transabdominal amniocentesis. They examined 124 pregnant women subjected during the 17th or 18th week of pregnancy to amniocentesis in the out-patient department. Before amniocentesis and soon after it they examined the flow in the uterine artery on the right and in the umbilical artery. From five consecutive cycles they evaluated the S/D ratio and RI in both vessels and compared the results. They did not record any complications during amniocentesis. Evaluation of the results of both investigated parameters before and after amniocentesis did not reveal any statistically significant differences in the investigated vessels. Based on the assembled results, the authors assume that transabdominal amniocentesis, despite its invasive character, does not influence the foetoplacental circulation and does not have a negative impact on the course of pregnancy.


[Determination of growth scales and ponderal indices in neonates].

By retrospective processing of data from 10,002 neonates the authors prepared tables of birth weights and heights incl. percentiles for the 28th-43rd week of gestation. By processing data concerning 9,893 neonates the authors elaborated ponderal indexes for the 32nd-43rd week of gestation. The tables make it possible to evaluate the nutritional status and proportionality of neonates.


[Transdermal administration of estrogens in women with the postcastration and climacteric syndromes].

The authors report on the results of a follow-up of 64 women aged 25-64 years, to whom oestrogens were administered transdermally by means of the preparation Estraderm TTS 25 or 50 for three months. 55 women were treated for postcastration syndrome and nine women for climacteric syndrome. The authors deal with the problems of dosage of the transdermal oestrogen therapy, study the side-effects of treatment and evaluate the results of therapy. In conclusion, they recommend the transdermal form of treatment by Estraderm TTS because of its very good therapeutic effect, acceptability by the patients, and only minimal local and general side-effects.


[Determination of the fertile period during the menstrual cycle in women by monitoring changes in crystallization of saliva with the PC2000 IMPCON minimicroscope].

The authors tested the possibility of assessment of fertile and infertile days during the menstrual cycle by investigating the crystallization of saliva by means of a minimicroscope PC 2000 IMPCON. They followed up for five months a total of 58 women where they assessed by a combination of at least three classical examination methods ovulatory and anovulatory cycles. They monitored a total of 120 cycles and examined 1649 specimens of saliva. Some women had to be eliminated on account of virosis during a flu epidemic (a total of 11 cycles). During the periovulatory period the authors recorded in ovulatory cycles the foreseen crystalline structures in saliva in 78.57%. In anovulatory cycles agreement between the expected character of the saliva specimen and anovulation was found in 84% of the examined cycles. In addition to structures suggesting fertile or infertile days, the authors identified also intermediary types. In the discussion the authors analyze various aspects as regards evaluation of the correlation between salivary and serum levels of gonadotropins and ovarian steroids. They evaluate physical and chemical changes of saliva and vaginal secretion and discuss the problem of crystallization of the saliva during the menstrual cycle. Based on their findings, when testing the minimicroscope PC 2000, they supplement the list of factors with influence the results of examinations. They consider PC 2000 a suitable modern equipment which extends the range of contraceptives and at the same time helps to assess the optimal time for conception in planned pregnancies.


[Sexuality in women after treatment of malignant breast tumors].

By means of a structured interview and four questionnaires (Heterosexual development of woman, Sexual activity of woman, Sexual function of woman and Questionnaire N5 which evaluates the presence and intensity of neurotic symptoms) the authors examined 154 women following treatment of a malignant breast tumour. During the examination which took place during spa treatment the age of the probands was 26-67 years. The most frequent adverse subjective sensations were problems of nakedness when seen by healthy women, when "seening themselves" and when seen by the husband (partner). Adverse feelings developed most frequently during the first week after operation. The adaptation was worst in women aged 40-44 years. Changes in emotional relations between partners were more favourable than in sexual life. Emotional as well as sex life improved in every fourth and more than in every 13th woman, as compared with the status before the disease. Men were quite tolerant. Fears and the degree of neuroticism corresponded with the deterioration of sexual function. Two-fifths of the women were interested in a reconstruction operation of the breast, however, only one in 40 of the group the operation was implemented or planned.


Construction and growth properties of a yeast strain defective in sterol 14-reductase.

We have transformed Saccharomyces cerevisiae with a genomic library contained in the replicative vector pFL44. The resulting transformants were screened for resistance to fenpropidin, a specific inhibitor of sterol 14-reductase. A plasmid was isolated that transformed yeast both to resistance to fenpropidin and to an increased specific activity of sterol 14-reductase. Sterol analysis of transformed cells grown in the presence of increasing concentrations of the inhibitor confirmed that resistance was a consequence of over-production of sterol 14-reductase. By chromosomal gene disruption, we have, for the first time, constructed yeast strains defective in sterol 14-reductase. As expected, since yeast in unable to take up sterols in aerobiosis, the disrupted strains do not grow in the presence of oxygen, even if exogenous sterols are supplied. However, disrupted cells grow in anaerobiosis with exogenous oleic acid and ergosterol supplements. They also grow in aerobiosis if they bear an additional mutation allowing sterol uptake. In this last growth condition the cells require a "sparking" ergosterol supplementation (25 nM) and accumulate ignosterol (ergosta-8,14-dienol) as the end-product of the sterol pathway. These results reveal that ignosterol is not obviously toxic to yeast membranes and strongly suggest that the molecular basis of the antifungal-activity morpholine and piperidine is directly related to the specific inhibition of ergosterol formation.


Biosynthesis of sulphur amino acids in Saccharomyces cerevisiae: regulatory roles of methionine and S-adenosylmethionine reassessed.

cys4-1, a mutation in the reverse trans-sulphuration pathway, relieves the sulphate assimilation pathway and homocysteine synthase from methionine-mediated repression. Since the mutation blocks the synthesis of cysteine from methionine downstream from homocysteine, this indicates that neither methionine nor S-adenosylmethionine serve as low-molecular-mass effectors in this regulatory system, contradicting earlier hypotheses.


The REV2 gene of Saccharomyces cerevisiae: cloning and DNA sequence.

The REV2 gene of Saccharomyces cerevisiae was cloned and sequenced; it contains an open reading frame of 1986 bp with a coding potential of 662 amino acids. Interruption of the chromosomal REV2 gene by integrating the URA3 gene coupled with partial deletion of the 3' terminal region produced viable haploid rev2 delta mutants. This indicates that the REV2 gene is non-essential for growth. The rev2 delta mutant is slightly more UV-sensitive than strains carrying various rev2 alleles (rev2-1, rev2x, rad5-1, rad5-8). The putative Rev2 protein is probably a globular protein containing a highly conserved nucleotide-binding site and two zinc-finger domains.


Genetic mapping of 1,3-beta-glucanase-encoding genes in Saccharomyces cerevisiae.

The map position of three 1,3-beta-glucanase-encoding genes in S. cerevisiae has been determined following conventional meiotic and mitotic mapping combined with recombinant DNA techniques. EXG1, EXG2 and SSG1 were localized to chromosomes XII, IV and XV, respectively, by hybridizing the cloned genes to Southern blots of chromosomes separated by pulsed-field gel electrophoresis, in conjunction with the rad52-1-dependent chromosome-loss mapping technique. Meiotic tetrad analyses further localized the EXG1 gene 6.1 centimorgans centromere-proximal to CDC25 on the right arm of chromosome XII. EXG2 was positioned between LYS4 and GCN2 on the right arm of chromosome IV, at distances of 6.2 centimorgans from LYS4 and 4.9 centimorgans from GCN2. Finally, the SSG1 locus mapped on the right arm of chromosome XV, about 8.2 centimorgans to the centromere-proximal side of HIS3.


Regulation of pho1-encoded acid phosphatase of Schizosaccharomyces pombe by adenine and phosphate.

Expression of pho1-encoded acid phosphatase of Schizosaccharomyces pombe has been reported to be regulated by phosphate. In this communication we show that it is also regulated by adenine. Starving adenine auxotrophic strains for adenine leads to a drastic increase of the enzymatic activity while adenine represses this activity. Full repression by adenine only occurs when phosphate is not growth limiting and vice versa. Regulation occurs at the level of mRNA. We isolated adenine non-repressible mutants. They define four genes (anr1, anr2, anr3, and anr5) which are involved in adenine-dependent pho1 expression. All anr mutants are also phosphate non-repressible. These results indicate that the generation and/or transduction of the intracellular signal responsible for pho1 repression is simultaneously dependent on both adenine and phosphate.


An NADP(+)-dependent glycerol dehydrogenase in Aspergillus nidulans is inducible by D-galacturonate.

In Aspergillus nidulans there is an NADP(+)-dependent glycerol dehydrogenase that is specifically induced on transfer to D-galacturonate medium. In contrast to the previously characterised constitutive NADP(+)-dependent glycerol dehydrogenase it has a much broader substrate specificity, having activity as an ethanol dehydrogenase, and is subject to carbon-catabolite repression. In addition to the two NADP(+)-dependent glycerol dehydrogenases, alcohol dehydrogenase I and II are also present on transfer to D-galacturonate medium, and have weak activity as glycerol dehydrogenases.


Structure and evolution of myxomycete nuclear group I introns: a model for horizontal transfer by intron homing.

We have examined five nuclear group I introns, located at three different positions in the large subunit ribosomal RNA (LSU rRNA) gene of the two myxomycete species, Didymium iridis and Physarum polycephalum. Structural models of intron RNAs, including secondary and tertiary interactions, are proposed. This analysis revealed that the Physarum intron 2 contains an unusual core region that lacks the P8 segment, as well as several of the base-triples known to be conserved among group I introns. Structural and evolutionary comparisons suggest that the corresponding introns 1 and 2 were present in a common ancestor of Didymium and Physarum, and that the five introns in LSU rRNA genes of these myxomycetes were acquired in three different events. Evolutionary relationships, inferred from the sequence analysis of several different nuclear group I introns and the ribosomal RNA genes of the intron-harbouring organisms, strongly support horizontal transfer of introns in the course of evolution. We propose a model that may explain how myxomycetes in natural environments obtained their nuclear group I introns.


Extrachromosomal ribosomal DNA of Didymium iridis: sequence analysis of the large subunit ribosomal RNA gene and sub-telomeric region.

The ribosomal DNA of the myxomycete Didymium iridis is organized as extrachromosomal linear molecules of about 20 kb, containing only one transcription unit of the ribosomal RNA genes. We have determined the sequence of the large subunit ribosomal RNA (LSU rRNA) gene as well as the sub-telomeric and telomeric regions. The LSU rRNA gene was found to encode a 3857 nucleotide-long LSU rRNA, interrupted by a transcribed spacer and two group I introns. A complete secondary structure model of D. iridis LSU rRNA has been constructed. The compact sub-telomeric region of D. iridis rDNA was found to contain several directly repeated sequence elements that include the simple telomere motif TTAGGG. Based on pairwise comparisons of LSU rRNA sequences, the time of divergence between the two myxomycete genera Didymium and Physarum was estimated.


Zinc fingerprinting for Phytophthora species: ZIF markers.

We have assessed the potential of using zinc finger markers for identification of fungal species using Phytophthora as a model organism, since it is particularly difficult to classify. The results show that such markers are suitable for species identification of Phytophthora but do not appear to aid strain identification within species.


Circular extrachromosomal DNA codes for a surface protein in the (+) mating type of the zygomycete Absidia glauca.

A small protein with a molecular mass of 15 kDa, which is specifically found on the hyphal surface of a (+) mating-type strain of the model zygomycete Absidia glauca, was purified to electrophoretic homogeneity and partially sequenced. The corresponding gene was cloned by means of an oligonucleotide probe deduced from the protein sequence. It could be localized on an extrachromosomal circular DNA element with a total length of 1250 bp. Electron microscopic analysis of A. glauca DNA showed that small extrachromosomal DNAs with varying length are a common feature of this zygomycete. There are no indications of additional chromosomal copies of the gene for this surface protein, and the plasmid is absent from DNA preparations of the (-) mating type. The copy number ranges around three per haploid genome, and a single transcript with a length of 400 bp, coding for the surface protein, could be found by employing a hybridization probe which spans the complete fungal plasmid. This is the first report of naturally occurring extrachromosomal DNA in a Mucor-like fungus, and the only example where an integral protein of the cell wall is encoded by a plasmid.


The plastome-encoded zfpA gene of a moss contains procaryotic as well as eucaryotic promoter consensus sequences and its RNA abundance is modulated by cytokinin.

Plastid DNA of the moss Physcomitrella patens has been sequenced. An open reading frame (ORF 315) was identified downstream from rbcL, between trnR-CCG and psaI. This ORF shares homology with zfpA, a putative regulatory gene in Pisum sativum. The moss ORF is preceded by a Shine-Dalgarno sequence, two plastid promoter consensus sequences, and three TATA boxes. A specific probe detected three transcripts of low abundance in the wild-type moss and a cytokinin-sensitive chloroplast mutant. Steady state levels of zfpA transcripts were different in the two genotypes. In mutant protonemata treated with cytokinin, steady state levels of the largest transcript decreased significantly.


A ten-minute protocol for transforming Saccharomyces cerevisiae by electroporation.

We present a simplified and rapid method for the transformation of yeast cells by electroporation. Stationary cells, scraped off the agar of Petri dish cultures stored in the refrigerator for up to 6 weeks, are suspended in sorbitol buffer, spun down by gentle centrifugation, transferred into the electroporation cuvette, and immediately subjected to transformation via electroporation. Transformation efficiency of this 10-min method, which does not require the preparation of cell cultures, is about 10% of the hitherto best performing transformation procedure using cells of defined growth phase.


High-level resistance to cycloheximide resulting from an interaction of the mutated pdr3 and cyh genes in yeast.

In addition to pdr3-1, the S. cerevisiae nuclear pleiotropic drug resistance mutant 2D was found to contain another recessive nuclear mutation, cyh, conferring specific resistance to cycloheximide only. The cycloheximide resistance level due to either the pdr3-1 or the cyh mutation alone was low and was not altered by the ogd1 mutation which increased the physiological acidification of the culture. When pdr3-1 and cyh mutations occurred simultaneously in the haploid yeast strain their interaction was synergistic and resulted in high-level resistance to cycloheximide.


In vivo conformation of mitochondrial DNA in fungi and zoosporic moulds.

Migratory behaviour of mitochondrial DNA (mtDNA) from fungal species belonging to Plectomycetes, Loculoascomycetes and the zoosporic moulds, Oomycetes, has been studied by Pulsed Field Gel Electrophoresis (PFGE). Electrophoretic profiles demonstrate that long, linear molecules of a heterogenous size are the prevailing in-vivo form of organelle DNA in all examined species. These profiles are consistent with the presence of the rolling-circle mode of mtDNA replication that occurs in all branches of true fungi and zoosporic moulds.


[The first stage of clinical trials of hydrogel lenses manufactured in Czechoslovakia].

The authors give an account of the results of the first stage of clinical trials of Czechoslovak disc-shaped hydrogel lenses made from the copolymer HEMA and methacrylic acid. Hitherto assembled experience provided evidence that the envelope technique for opening of the anterior capsule does not ensure sufficient stability of these lenses in the later stage and their decentration may cause serious complications. Therefore the optimal solution is to place the implant in the capsule after circular capsulorhexy. Improvement of the geometrical and optic properties of the lenses during the subsequent follow-up and their perfect harmony with the surgical technique will produce even better functional results in operated patients.


[A method for determining peroperative centering and monitoring the development of postoperative decentration of intraocular lenses].

After implantation of intraocular lenses (IOL) Purkinje images 3 and 4 (Pi3 and Pi4) are clearer on the anterior and posterior surface of its optical part (subsequently only lens). Along with the corneal Pil this phenomenon was already used in clinical work for more accurate calculations of the degree of decentration and tilt of the lens in the eye. The authors describe a simplified method for assessment of the position of the lens by means of Pil, 3 and 4, using apparatuses with an approximately coaxial light. The method uses overlapping of Pi1, 3 and 4 in perfect centering of the lens in the axis of the eye (it is assessed by drawing a perpendicular line on the centre of the cornea) and marked dislocation of Pi3 in the direction of decentration of the planoconvex lens with the convexity facing the cornea. The inclination of the lens must be eliminated according to positions Pi1 and Pi4. The method is suitable for peroperative centering and for assessment of the direction and extent of decentration of these lenses. The authors discuss the possible use of this method also in biconvex and planoconvex lenses with the convexity facing the retina. In a group of 10 implanted planoconvex lenses the authors demonstrate the possibility of perfect peroperative centring of the lens and other applications of the method. Incorrect position of the IOL at the end of the operation, which can be one of the factors of later complications, did not change during the early postoperative period.


[Analysis of the first 1000 implants of intraocular lenses].

The authors give an account of the development of implantology and extracapsular extraction in their department. In 1987 they performed 342 operations of cataract, an intraocular lens was implanted in 13.2%, extracapsular extraction was performed in 12.9%. In the first half of 1991 354 operations of cataract were performed, an intraocular lens was implanted in 82.5%, and extracapsular extraction was performed in 96.9%. The most frequent peroperative complication was rupture of the posterior capsule--from 14.0% to 20.1%. The most frequent postoperative complication (306 implantations in the first half of 1991)--keratopathia striata 179, fibrinous exudate--79, elevated intraocular pressure--50. The time spent in hospital in the first half of 1991 was on average 2 days in 156 operated patients who did not develop any or only one postoperative complication. The most frequent postoperative complication was posterior capsule opacification--11%. The most serious late complication during the entire follow-up period was endo-ophthalmitis in 8 eyes which in 5 patients after an interval up to 12 months following operation ended by evisceration of the bulbus--in all instances Russian lenses which were sterilized by ourselves were involved. The visual acuity on admission was below 6/60 in 72.3%, on discharge 6/12 or better in 32.8% and 6 months after operation 6/12 or better in 73.9%, incl. 651 eyes (88%) without supplementary correction. Despite the difficult conditions extracapsular extraction with implantation of an intraocular lens has become a routine method in our department, with a short period of hospitalization and minimal contraindications.


[The individual A constant in the SRK II formula].

The authors investigated the postoperative dioptric deviation from the calculated optic power of emetropic intraocular lenses by means of the formula SRK II without and with use of the individual regression constant A. Within the range of +/- 1.5 D the deviation varied for both methods in 87.7% of operated eyes; between -1.0 and +1.0 D the percentage of dioptric deviations without personalization was 66.2 and with the use of the individual constant A it was 73.8.


[Determination of correction based on natural visual acuity in myopes].

The authors demonstrate the possibility to assess the correction in ametropes according to the value of their natural vision. To this end they use normalized optotype tables with Landolt rings which have a constant diminution coefficient between individual consecutive lines. By means of this elaborated method the authors were able to assess in twenty myopic subjects that a correction of -1.0 corresponds to an improvement by four lines, one line thus corresponds to a correction of -0.25 dioptres.


[Isolation of Dirofilaria repens in vitreoretinal findings].

The authors present the case of a 18-year-old patient who reported deteriorated vision and mobile dark shadows on the left eye. Examination revealed in the vitreous body an extremely mobile filiform, worm-like formation, white in colour, of the size of an arteriole, about 9 mm long. In the course of hospitalization the formation was isolated by surgery of the pars plana vitrectomy and subjected to detailed helminthological examination. The worm was identified as Dirofilaria repens (Reillet et Henry 1911) and described in detail. The presented finding is the first finding of this parasite in this country and at the same time the first finding of this parasite in Czechoslovakia.


[Retinal detachment with retinal dialysis].

A group of 165 eyes operated on account of detachment of the retina comprised 16 eyes with oral dialysis (i.e. 9.7%). Only in 6 eyes of the group there was a positive traumatic case-history. The anatomical results of the operations were favourable (81% amotions cured), the functional results depended on the state of the macula before operation.


[5 years' experience in the treatment of neovascular glaucoma using cryocoagulation. I. Development of intraocular pressure].

The authors evaluate the results of reduction of intraocular pressure in 38 eyes with neovascular glaucoma after a cryosurgical operation. In 12 eyes they used cyclocryocoagulation as an independent method, in 26 eyes transscleral panretinal cryocoagulation combined with cyclocryocoagulation in one session. The achieved results indicate, that intraocular pressure in neovascular glaucoma can be influenced more readily by a combined cryosurgical operation than by cyclocryocoagulation alone. When using transscleral panretinal cryocoagulation in combination with cyclocryocoagulation, they achieved after operation an intraocular pressure under 26 mm Hg within 3 days after operation in 50%, within 10 days in 76.9%. They observed a subsequent rise of the intraocular pressure in 15%. Hypotension developed in 30.8%. A quite high percentage of eyes with extreme hypotension and subsequent atrophy of the bulbus with amourosis (27%) is according to the authors due to too extensive cyclocryocoagulation.


[5 years' experience in the treatment of neovascular glaucoma using cryocoagulation. II. Treatment of pain and the development of visual acuity].

The authors evaluate the therapeutic results of pain of the eyes and the development of visual acuity of 38 eyes with neovascular glaucoma treated by cryocoagulation. In 12 eyes they used cyclo-cryocoagulation as an independent method and in 26 eyes the method of transscleral panretinal cryocoagulation combined with cyclo-cryocoagulation. The authors did not find a correlation between the degree of increased intraocular pressure and the painfulness of the eye. In the treatment of pain they achieved markedly better results when using transscleral panretinal cryocoagulation than cyclo-cryocoagulation alone. The better therapeutic results of treatment of pain by combined cryosurgery is associated with a better compensation of intraocular pressure. In the authors' view a significant in the treatment of pain is also played by the newly described so-called analgetic effect of panretinal cryocoagulation which develops probably as a result of damage of sensitive nerves of the bulbus during freezing of the sclera and the choroid. The development of the so-called analgetic effect is probably associated with two facts: relief from severe pain was perceived also by patients where during the early postoperative period a high intraocular pressure still persisted and also in patients where a high intraocular pressure rose again. This condition was, however, as a rule not associated with the onset of new pain. The use of transscleral panretinal cryocoagulation combined with cyclo-cryocoagulation achieved regression of pain within 5 days after operation in 100%. The author provide evidence that the visual acuity in neovascular glaucoma recedes very rapidly and irreversibly.(ABSTRACT TRUNCATED AT 250 WORDS)


[Use of 5-fluorouracil in complicated cases of glaucoma--personal experience].

The authors give an account of the results of trabeculectomy with the use of 5-fluorouracil in a group of 8 patients with complicated glaucoma. Treatment was successful in 5 patients, in three patients it failed. The use of 5-fluorouracil in specific cases of glaucoma gives greater hope of successful surgery.


[Rhabdomyosarcoma of the orbit].

A twenty-eight year-old female patient developed within one week a protrusion and deviation of the eyeball associated with pain and loss of vision. The sudden development, atypical course, obscure clinical picture suggest rather an inflammatory or vascular affection in the orbit; only subsequent development confirms the development of a malignant process in the orbit.


[Therapeutic and cosmetic effect of cryocoagulation therapy of basaliomas in the area of the eye].

The authors evaluated the results of cryosurgical intervention in a group of patients with advanced basaliomas with surgically adverse localizations from the aspect of relapses and the cosmetic effect. Due to the simple procedure and achieved effect this technique is a considerable asset.


[Betoptic, a new possibility in the treatment of glaucoma].

The authors present an account of their findings in connection with a new selective beta 1 adrenergic antagonist in the preparation Betoptic. They compared its effect on intraocular pressures, pulse rate, systemic blood pressure and vital capacity of the lungs in ten patients with primary open-angle glaucoma with the effect of Timoptol and placebo in equally sized groups. They found that the preparation reduces reliably the intraocular pressure, it does not reduce the pulse rate and does not influence the vital capacity and systemic blood pressure. They did not detect any local or general undesirable effects. It can be therefore used even in subjects where unselective beta blockers are contraindicated or poorly tolerated.


[Surgical treatment of strabismus in children (a 12-year study)].

A group of 2205 operations of strabismus in the course of 12 years reveals a clear predominance of operations of dynamic strabismus (94%), as compared with surgery of paralytic strabismus and ocular torticollis on account of nystagmus (6%). This fact provides evidence of a marked ratio of a non-paralytic aetiology of strabismus in the child population. In esotropia, the most frequent type of strabismus, the authors consider as most suitable the technique of weakening of the inner rectus muscles by a dosed elongation according to Gonin-Hollwich, as compared with the classical retroposition of this muscle. In exotropia the authors recommend reinforcing operations only or in combination with a weakening operation of the rectus muscles. The gradual development of application of the technique of surgery of the hyperfunctional lower oblique muscle is in favour of treble partial myotomy (elongation). They operate paretic strabismus when the IIIrd, IVth, VIth nerve are affected and supranuclear paresis of the levators by a complex procedure incl. transposition operations of the functional muscles. The authors operate ocular torticollis after a careful analysis of the congenital nystagmus, using special techniques on the rectus and oblique muscles which adjust the position of the head and bulbs.


[Bone age of preschool children in Eastern Slovakia].

The authors present the results of evaluation of the skeletal maturation in 579 children aged 1-5 years. Bone maturation was evaluated, based on skeletal age, by the Tanner-Whitehouse II method (Tanner et al., 1975). It was found that girls up to the age of three years had lower and in the more advanced age groups higher values of skeletal age than boys. Boys were retarded as regards skeletal age in relation to chronological age on average by 0.28 years and girls by 0.25 years.


[Personal experience with genetic study of the hearing-impaired].

In 1988-1990 the authors examined on account of impaired hearing in the family 78 families, which were divided into two groups. The first group comprises 48 families of actively invited pupils from schools with children with impaired hearing in Prague, the second group comprises 30 families examined in the framework of common counselling and consultation services. On the first group a genetic aetiology of the disorder was revealed in 75% and in the second group in 73.3% of the cases. An autosomal recessive type of heredity was proved unequivocally in 31.3% in group 1 and in 20.0% in group 2. An autosomal dominant type of heredity was found in 6.25% in group 1 and in 6.7% in group 2. X-linked deafness was proved only in group 1 in 4.2% and sporadic syndromes only in group 2 in 13.3%. In 33.3% in both groups it was not possible to differentiate the type of heredity (most frequently the child and both parents were affected). In the first group three genetic syndromes were detected and in group 2 seven genetic syndromes. As the percentage of genetically conditioned hearing disorders in both groups is relatively high, the authors consider active screening of deaf children more useful so far.