Lymphocytes from multi-transfused patients exhibit cytotoxicity against autologous cells. We previously demonstrated that multitransfused patients with severe aplastic anaemia (SAA) exhibit high numbers of alloreactive cytotoxic T lymphocyte precursors directed against their HLA identical siblings. In this study a group of patients who had received multiple blood transfusions for SAA, other haematological diseases or acute blood loss were tested for autocytotoxicity and the results compared with those of untransfused controls. These controls consisted of normal individuals, patients with chronic myeloid leukaemia (CML) or untransfused patients with SAA. There was a significantly higher degree of autocytotoxicity in multitransfused patients, than in the untransfused controls, including untransfused patients with SAA (P = 0.0001). These results suggest that blood transfusion is responsible for inducing autoreactivity. In one patient, in whom both alloreactive anti-non-MHC and autoreactive cytotoxic T lymphocytes (CTL) had been detected, it was demonstrated that there was no crossreactivity between the alloreactive and autoreactive CTL responses. Inhibition studies using monoclonal antibodies revealed the effector cells to be T lymphocytes and the restricting determinants to be both HLA class I and II molecules.


Epidermolysis bullosa acquisita induced by GM-CSF: a role for eosinophils in treatment-related toxicity. A patient treated with granulocyte-macrophage colony-stimulating factor (GM-CSF) developed eosinophilia and epidermolysis bullosa acquisita. The bullae were subepidermal, and filled with an inflammatory infiltrate composed predominantly of eosinophils. Immunofluorescence studies disclosed linear deposition of IgG, IgA and C3 at the basement membrane zone and immunoelectron microscopy demonstrated antibody deposition in the lamina densa and sublamina densa region; however, the patient's serum did not contain circulating antibody to basement membrane zone antigens. Staining with monoclonal antibodies revealed dense deposits of both eosinophil peroxidase and eosinophil major basic protein at the dermal-epidermal junction. The eosinophilia and skin lesions resolved upon discontinuation of GM-CSF. This case provides evidence for two hypotheses: (1) GM-CSF induced proliferation and activation of eosinophils may contribute to some of the toxicities of GM-CSF treatment, and (2) activated granulocytes, including eosinophils, may mediate blister formation in epidermolysis bullosa acquisita.


Remission with morphological myelodysplasia in de novo acute myeloid leukaemia: implications for early relapse. Myelodysplastic features of remission bone marrow were investigated in 46 adults with de novo acute myeloid leukaemia (AML) according to the FAB morphological criteria for myelodysplastic syndromes (MDS). Compared with a group of 18 patients with the common type of acute lymphoblastic leukaemia in remission, micromegakaryocytes (30.4% v. 5.6%, P less than 0.05), multi-separated nuclear megakaryocytes (45.7% v. 0%, P less than 0.01), degranulated neutrophils (39.1% v. 5.6%, P less than 0.05), and neutrophils with hyposegmented nuclei (34.8% v. 0%, P less than 0.01) were significantly more common in the AML patients. In contrast, dyserythropoietic changes had a similar incidence in both groups. When compared with the clinical features at initial diagnosis in AML cases, the dysmegakaryocytic changes in remission marrow were found to be significantly more frequent in patients with monocytic involvement (mainly M4) or trilineage myelodysplasia (T-MDS AML). Disease-free survival was significantly shorter in patients with micromegakaryocytes (P less than 0.05) or neutrophils with hypogsegmented nuclei (P less than 0.05) than in those without these features. Overall survival was also significantly shorter in patients with micromegakaryocytes (P less than 0.05). These findings may help in developing new strategies for the post-remission therapy of AML, and also suggest that myelodysplastic changes in the remission marrow of de novo AML patients may be related to haematopoietic recovery by a preleukaemic clone which eventually leads to early leukaemic relapse. Dysplastic marrow, however, was not necessarily associated with peripheral pancytopenia in the present series, warranting basic research on such putative clonal remissions.


Immunophenotypic subclassification of chronic lymphocytic leukaemia (CLL). To determine the significance of the immunophenotypic heterogeneity of B-cell chronic lymphocytic leukaemia (CLL), surface immunoglobulins (SIgs), mouse rosette assays (MR), and a panel of monoclonal antibodies for B cells, T cells and myeloid cells were performed on peripheral blood samples from 61 newly diagnosed cases. Four groups were observed: group I (SIg+, MR+, CD19/20+, CD5+, T antigen (Ag)-; 27 cases); group II (SIg+, MR+, CD19/20+, CD5+, T Ag+; 17 cases); group III (SIg+, MR+ CD19/20+, CD5-, T AG-; 12 cases); and group IV (SIg-, MR+, CD19/20+, Cd5+, T Ag-; 5 cases). Groups were compared according to French-American-British Cooperative Group subtypes, clinical and laboratory features, Rai staging, and survival. Typical CLL morphology (greater than 90% small lymphocytes) was present in 20/20 (100%) of group I cases and 23/27 (85%) group II, III and IV cases (P = 0.09). Expression of a myeloid antigen was seen in 5/27 group I cases (18%) and 1/16 group II cases (6%), but was not predictive of survival (P = 0.36). The CD5- group III had a lower haemoglobin level (P less than 0.0001), higher Rai stage (P less than 0.002), and poorer survival at 5 years (P less than 0.02) than the other groups. We conclude that at least four distinct immunophenotypic subgroups of B-cell CLL can be determined. Expression of myeloid or T-cell antigens does not appear to predict for patient survival; however, lack of CD5 antigen may be associated with more advanced stage of disease and poor patient survival.


Cytomorphology of childhood lymphoblastic leukaemia: a prospective study of 2000 patients. United Kingdom Medical Research Council's Working Party on Childhood Leukaemia. Blast cell morphology of children with lymphoblastic leukaemia (ALL) entering two national multicentre trials was prospectively reviewed by three haematologists to define the clinical importance of (a) French-American-British (FAB) classification, (b) the presence of cytoplasmic vacuoles, and (c) the presence of 'hand-mirror' cells. Of 2135 evaluable children, 1907 (89%) had FAB L1 morphology and 228 (11%) L2. (L3 patients were not eligible for the trials in question). L2 patients more frequently had residual disease 14 d after starting treatment and had a significantly inferior disease-free survival, but not if the analysis was stratified for age, sex and diagnostic white cell count (WBC). 627 (29%) had blast cells with cytoplasmic vacuoles, and showed a significant survival advantage over the remainder. Vacuoles were positively associated with a low WBC, age range 1-6 years and blast cell positivity for CD10, but their benign influence was apparent even when these variables were taken into account. 'Hand-mirror' (HM) cells were only studied in UKALL X, and were noted in 316/1402 (23%) children. There appeared to be an inverse correlation between HM cells and cytoplasmic vacuoles and a weak association with T-cell immunophenotype, but no prognostic significance was evident. FAB classification appears to be of less prognostic importance than has previously been supposed, though L2 disease is more resistant to current remission induction regimens. Hand-mirror cells may be more common in T-ALL, but are seen in all types and are not related to prognosis. Cytoplasmic vacuoles are predictive of a good response to current therapeutic schedules even allowing for other prognostic variables, and are the single most important morphological feature relating to prognosis in childhood ALL.


Comparison of chromosome analysis and BCL-1 rearrangement in a series of patients with multiple myeloma. Translocation (11;14)(q13;q32) is a recurring chromosome abnormality which is found non-randomly in non-Hodgkin's lymphoma, especially of follicular type, as well as in B-cell chronic lymphocytic leukaemia, Waldenström's macroglobulinaemia and multiple myeloma. To define further the prevalence of this abnormality in multiple myeloma, we studied a series of 17 patients with this disease with concomitant chromosome analysis and Southern blotting, using a probe specific for the major translocation cluster of the BCL-1 oncogene which is located at chromosome 11q13. Karyotype analysis of 14 evaluable patients revealed three cases with abnormalities of chromosome 11q13, two of them with t(11;14)(q13;q32) and one with del (11)(q13). Southern blot analysis showed no rearrangements in BclI and HindIII digests of DNA from 17 patients including the three patients with anomaly of chromosome 11q13, using a BCL-1 specific probe. A possible restriction length polymorphism was detected in EcoRI cut DNA of five out of 11 patients studied. Therefore the chromosomal break point in our cases with abnormality of chromosome 11q13 must lie outside the major translocation cluster of the BCL-1 gene. However, in centrocytic lymphoma rearrangement of the BCL-1 oncogene has been detected in up to 50% of cases. Location of the break point may therefore be dependent on the differentiation of the transformed B-cell.


Interleukin-4 and nerve growth factor can act as cofactors for interleukin-3-induced histamine production in human umbilical cord blood cells in serum-free culture. In order to better define the role of recombinant interleukins and growth factors in supporting histamine production by histamine-producing-cells (HPC), namely basophils or mast cells, we have conducted experiments in which we compared the abilities of IL-3, IL-4, IL-6 and nerve growth factor (NGF), either alone or in various combinations, to stimulate histamine production from human umbilical cord blood mononuclear cells cultured in either the presence or absence of serum. Over a period of 15 d, IL-3 was able to support intracellular histamine production, in both serum-supplemented and serum-free medium, although the effectiveness of IL-3 was lower in the serum-free cultures. IL-4, NGF and IL-6 taken individually, exhibited little, if any, histamine-promoting activity regardless of whether or not the cells were cultured in serum. Moreover, none of these cytokines significantly enhanced the effect of IL-3 in serum-supplemented cultures. In contrast, IL-3-promoted histamine production in serum-free cultures was significantly enhanced by IL-4 and NGF. These results demonstrate that both IL-4 and NGF can function as cofactors with IL-3 in the support of histamine production in human umbilical cord blood cells.


Myelodysplastic syndrome (MDS)-associated inhibitory activity on haematopoietic progenitor cells: contribution of monocyte-derived lipid containing macrophages (MDLM). We studied myelodysplastic syndrome-associated inhibitory activity (MDS-IA), which inhibited colony formation in vitro of normal granulocyte-macrophage progenitors (CFU-GM). When adherent marrow cells were incubated with fetal calf serum for 21-24 d, monocyte-derived lipid containing huge macrophages (MDLM) developed. MDLM from MDS marrow (MDS-MDLMs) and their conditioned medium (MDLM-CM) consistently suppressed the growth of normal CFU-GM colony formation. MDS-IA was active on CFU-GM during the S-phase and relatively resistant to heating. Monoclonal antibody against H subunit (acidic) ferritin and polyclonal antibody against placental ferritin neutralized the inhibitory activity of MDS-MDLMs. In addition, cell lysates of MDS-MDLMs reacted to both monoclonal anti-H subunit ferritin and polyclonal anti-placental ferritin in Western blotting analysis, indicating that the inhibitory activity was predominantly acidic isoferritin. On the other hand, MDLMs obtained from normal bone marrow had a CFU-GM enhancing activity. These results suggest that MDS-MDLMs may be responsible for the suppression of granulopoiesis in patients with MDS and that the suppression may be mediated by soluble factors, notably H subunit isoferritin.


Ultraviolet irradiation modulates MHC-alloreactive cytotoxic T-cell precursors involved in the onset of graft-versus-host disease. Ultraviolet B (UVB) irradiation of cellular blood components has been proposed as a new technology to prevent HLA sensitization in recipients. Earlier studies have shown that a dose of 2 J/cm2 abrogates the ability of lymphocytes to serve as stimulators in mixed lymphocyte cultures (MLC). In this study we have evaluated the effect of UV energy on T-lymphocytes for the prevention of transfusion-associated graft-versus-host disease (TA-GvHD). The response of cytotoxic T-lymphocyte precursors against host alloantigens was almost undetectable at a dose of 0.5 J/cm2. T-cell proliferation in MLC or in response to phytohaemagglutinin was inhibited by more than 95% at doses of 1 J/cm2 or higher. The data suggest that UV irradiation can be used to prevent both HLA sensitization and TA-GvHD in recipients.


Philadelphia positive acute leukaemia with minor breakpoint cluster rearrangement may be a stem cell disease. The biology of Philadelphia chromosome positive (Ph+) acute lymphoblastic leukaemia (ALL) is the subject of much interest. We present a case of Ph+ ALL with a minor breakpoint cluster (mBCR) rearrangement who subsequently relapsed with Ph+ mBCR+ acute myeloid leukaemia and later with Ph+ mBCR+ acute stem cell leukaemia. This case provides further evidence that Ph+ ALL with a mBCR rearrangement may arise from a pluripotent stem cell with similar potential to that of chronic granulocytic leukaemia to undergo blastic crises with differing lineage characteristics.


Specific binding between human neutrophils and heparin. Heparin binding on polymorphonuclear leucocytes (PMNL) was characterized. Heparin binding was specific, rapid, saturable and reversible. One single class of heparin binding sites was found with a dissociation constant of 1.22 mumol/l and 7.7 x 10(6) sites per PMNL. The binding was independent of the anticoagulant activity of heparin. Heparin affinity chromatography on radio-iodinated cell lysates followed by polyacrylamide gel electrophoresis in the presence of sodium dodecylsulphate revealed a 130 kD heparin binding protein. Heparin binding was inhibited by disodium salt of ethylenediamine tetraacetic acid. Cell surface bound heparin was functionally inactive and did not affect the inactivation of thrombin by antithrombin III. Our study demonstrates that heparin interacts with PMNL by a cell-surface binding protein. These instructions could be consistent with the modifications of some PMNL functional properties in the presence of heparin.


A family with haemolytic anaemia and three beta-globins: the deletion in haemoglobin Atlanta-Coventry (beta 75 Leu----Pro, 141 Leu deleted) is not present at the nucleotide level. Analyses of haemoglobin from a family with an unstable haemoglobin haemolytic anaemia demonstrated that the affected individuals had three beta-globins, namely, normal (beta A), Atlanta (beta At) with a mutation of beta 75 Leu----Pro, and beta-Atlanta-Coventry (beta At-Co) with mutation of beta 75 Leu----Pro and beta 141 Leu deleted. These were present in the ratio 66:23:11 respectively. The structure of the beta-globin cluster, however, was found to be normal by Southern blotting; also cytogenetic analysis failed to show any abnormality. DNA sequence analyses demonstrated the presence of the beta At mutation in genomic DNA isolated from leucocytes but the Coventry deletion of 141 Leu in beta At-Co was not present in genomic DNA. PCR amplification of the beta-globin cDNA and direct sequencing of the product also failed to demonstrate the Coventry deletion. Thus, it appears that the absence of 141 Leu in the beta At-Co globin is a consequence of the beta At mutation in these patients and that both beta At and beta At-Co are the product of a single gene. This unusual conclusion is paralleled in the bizarre case of Hb Vicksburg where the deletion of a leucine at beta 75 is not coded for in genomic DNA.


Beta 141 Leu is not deleted in the unstable haemoglobin Atlanta-Coventry but is replaced by a novel amino acid of mass 129 daltons. Reinvestigation of the structure of the beta-chain of Hb Atlanta-Coventry (beta 75 Leu----Pro, beta 141 Leu deleted) confirmed the presence of two abnormalities; however, analysis of the aberrant beta Co14 tryptic peptide by liquid secondary ion mass spectrometry indicated that the beta 141 Leu (mass 113 daltons) was not deleted but replaced by a novel amino acid of mass 129 daltons. The new amino acid in peptide beta Co14 was uncharged at pH 6.5, more hydrophillic than leucine and susceptible to cleavage by both chymotrypsin and carboxypeptidase A. We propose that the new residue is likely to be hydroxyleucine and that it results from post-translational oxidation of beta 141 Leu as a consequence of perturbation of the haem environment caused by the beta 75 Leu----Pro mutation in the E helix (E19). This proposal is entirely consistent with recent DNA analysis which showed that beta At-Co was not the product of a third beta-globin gene and that neither of the two beta-globin genes, beta A nor beta Atlanta, contained a deletion of the beta 141 Leu codon. We have subsequently found this modified amino acid at position beta 141 in two other unstable haemoglobins, both of which involve mutations on the haem side of the E helix.


A rapid solid-phase extraction and HPLC/DAD procedure for the simultaneous determination and quantification of different benzodiazepines in serum, blood and post-mortem blood. A rapid and quantitative method for the determination of benzodiazepines using high-performance liquid chromatography (HPLC) with diode-array detection (DAD) is reported. The drugs were extracted from serum, blood or post-mortem blood using C18 extraction columns. Brotizolam was used as internal standard. Experiments with spiked serum/blood samples resulted in recoveries between 75% and 94% for all investigated benzodiazepines. Excellent linearity was obtained over the concentration range 5-1500 ng benzodiazepine/ml. The limit of detection was approximately 2 ng/ml. The detection of low therapeutic serum levels of highly potent benzodiazepines is also possible.


The elimination kinetics of methanol and the influence of ethanol. Four male subjects aged between 20 and 29 years were given intravenous injections of methanol at a dosage of 10 mg per kg body weight, once without prior administration of ethanol, and once after oral ingestion of 0.3 g ethanol per kg body weight. The serum methanol concentration was monitored over the next 5 h (after methanol administration alone) and 6-7 h (after methanol administration following ethanol ingestion). The elimination of methanol administered alone was found to follow first-order kinetics with a rate constant for the elimination phase of 0.475-0.259 h-1, corresponding to an elimination half-life of 1.8-3.0 h. When ethanol was also administered methanol oxidation was found to be completely blocked until the blood ethanol concentration had fallen to 0.2 g/kg. When the ethanol concentration had dropped to zero, methanol elimination followed exactly the same course as that observed in the experiment without prior administration of ethanol (k: 0.378-0.231 h-1; t1/2: 1.5-2.7 h).


The screening for common drugs of abuse in whole blood by means of EMIT-ETS and FPIA-ADx urine immunoassays. The purpose of the paper was to compare the performance of ETS (EMIT) and ADx (FPIA) analyzers for screening blood samples for drugs of abuse after 2 alternative pretreatment procedures (acetone precipitation and ultrafiltration). Cannabinoids, benzodiazepines and benzoylecgonine were not detectable with both assays after ultrafiltration. The detectability of morphine in blood ultrafiltrates was distinctly lower than after acetone precipitation. The comparison of results obtained with ETS and ADx after acetone precipitation showed that ETS assay is slightly more sensitive but ADx is slightly more reproducible. Both assays may be used for blood examination with similar cut-off values. The ETS analyzer gave much better analytical performance (speed, flexibility) and lower reagent costs than the ADx analyzer.


Lethal myocardial infarction subsequent to compression of the left anterior descending coronary artery induced by traumatic hematoma. Post-traumatic myocardial infarction is a well-known although rare complication of blunt chest traumas. In cases of proximally located damage to the coronary arteries, modern vascular surgery such as bypass operations may be lifesaving. In this paper we describe a case, where a healthy 35-year-old man developed a lethal myocardial infarction 8 days after a chest trauma caused by a moped. The ECG obtained 4 h after the accident showed extensive transmural ischemia of the anterior wall. At autopsy a hematoma originating from a side branch was found to compress the left anterior descending coronary artery immediately distal to its origin from the main stem. This finding suggests that bypass surgery could have saved the life of this young man. Patients with evidence of myocardial ischemia following chest traumas should be transferred to a cardiac center as early as possible.


Evaluation of sperm specific lactate dehydrogenase isoenzyme C4 (LDH C4)--application to semen detection in stains. The usefulness of LDH C (or LDH X) as a semen-specific marker has been tested. Many different methods have been applied for identification using various electrophoretic methods such as agarose gel electrophoresis, IEF, rocket-immunoelectrophoresis with subsequent enzymatical, immunological and immunochemical detection methods. Further methods applied were dot blot analysis for LDH C and PCR analysis for X/Y-chromosome differentiation. In blood/semen mixtures the LDH C-bands were detectable in dilutions up to 50-fold after staining for total LDH activity. An appropriate extraction procedure was developed. Immunological tests using a polyclonal antibody against LDH C were found to be highly specific and highly sensitive. The immunological test gave positive results with azoospermic stains up to a few weeks and in normal sperm stains positive reactions were obtained even after 8 months storage. A comparison of different detection methods for semen showed that immunological detection methods for LDH C are, at least after longer time periods of storage, superior to microscopical detection. Swabs showed positive LDH C-reactions up to 48 h after intercourse. This reaction was closely correlated to PCR for male cells and to microscopical searching.


Interpretation of COHb concentrations in the left and right heart blood of cadavers. Carbon monoxide hemoglobin (COHb) concentrations in left and right heart blood samples from cadavers both exposed and not exposed to fire or CO gas were analyzed by the gas chromatographic method. The COHb concentration ratio between samples of left and right heart blood (L/R ratio) does not appear to be useful for establishing whether death has occurred before or after exposure to fire with the exception of cases where no soot can be detected in the airways by the naked eye and the COHb concentration in the blood sample is within the level considered normal for tobacco smokers.


Nerve conduction as a means of estimating early post-mortem interval. Methods in current practice for ascertaining time of death are largely based on the cooling of the body after death and are somewhat unreliable. A theoretical relationship is known to exist between the decline in the properties defining nerve conduction and time after death caused by the gradual cessation of metabolic activity in nerves. A number of such properties were measured in rats during life and after death. In most cases the relationship was found to be inconsistent. The chronaxie of the strength duration curve for the sciatic nerve was, however, found to increase consistently and reproducibly in a linear fashion over the first 90 min after death to a plateau value which was maintained beyond 135 min. These findings are discussed as the possible basis of a forensic method of determining the duration of the "post mortem interval" within the first few hours after death.


ABH-related antigens in human male genital tract. A histochemical examination. The localization of ABH related antigens in human male reproductive tract was examined using monoclonal antibodies and an avidin biotin complex method. No positive reaction with blood group antibodies on spermatozoa was observed in testis and ductus epididymidis apart from erythrocytes and endothelial cells. The expression of ABH and ABH related antigens in ductuli efferents testis, ductus epididymidis, seminal vesicle and prostate was complexly coded by a combination of H, Se, Le and X genes. The results obtained in this study indicate that the ABH antigens detected on spermatozoa of seminal stains are coating antigens and not inherent to the cell membrane, and the ABO, H, Se, Le and X genes are subjected to a tissue-dependent differential expression.


Genetic polymorphism of human alpha 2 HS-glycoprotein (AHSG) in the resident population of the Basque Country (northern Spain). The genetic polymorphism of human alpha 2 HS-glycoprotein (AHSG) was studied in a sample of 466 healthy unrelated individuals resident in the Basque Country (Northern Spain) by isoelectric focusing on micro-ultrathin polyacrylamide gels followed by immunoblotting. The allele frequencies obtained were AHSG*1 = 0.7253, AHSG*2 = 0.2683 and AHSG*3 = 0.0064. These allele frequencies were compared with those reported in other European populations.


Identification of human remains by amplification and automated sequencing of mitochondrial DNA. The highly decomposed remains of a corpse were identified by the amplification and direct sequencing of mitochondrial (mt) DNA. Degraded DNA was extracted from bone fragments and a necrotic skin sample and amplified at 2 hypervariable segments within the mitochondrial non-coding region using 2 rounds of nested PCR. Both strands of the amplified regions were sequenced and compared with each other to ensure fidelity of the data. Sequences from the bone and skin were found to be identical and matched data generated from a blood sample provided by a presumptive sister of the deceased. Thus, this evidence was consistent with the sister and the deceased being related.


Diffusion as a mechanism of postmortem drug redistribution: an experimental study in rats. In some cases of drug overdose there is a reservoir of unabsorbed drug in the stomach and gut. Furthermore, agonal aspiration might establish a second reservoir in the lungs. Two experimental rat models were used to study if diffusion from these reservoirs could contribute to the phenomenon of postmortem drug redistribution. Overnight fasted rats were sacrificed by CO2 and 75 mg of amitriptyline (AMI) was administered by a gastric tube. In the first series (n = 19), the tubes were removed after AMI administration. In the second series (n = 17), the trachea was ligated and cut prior to drug administration to prevent airways contamination. The rats were left at room temperature on their back for a period of 5, 10, 24, 48, 96 up to 192 h and samples of heart blood, blood from the inferior vena cava, tissue samples from heart, lungs, different liver lobes, kidney and psoas muscle were taken. In both series of rats we observed that as early as 5 h postmortem increasing concentrations of amitriptyline were found in the liver lobes lying closest to the stomach. In rats where the trachea was not ligated, drug contamination of the lungs also resulted in an increase in drug concentration within 5 h in heart blood and heart muscle. In rats where the trachea had been ligated, amitriptyline was found in the lungs after 96 h postmortem. The main metabolite nortriptyline was also detected.(ABSTRACT TRUNCATED AT 250 WORDS)


The time-dependent rearrangement of the epithelial basement membrane in human skin wounds--immunohistochemical localization of collagen IV and VII. In 62 human skin wounds (surgical wounds, stab wounds and lacerations after surgical treatment) we analyzed the immunohistochemical localization of collagen IV in the epithelial basement membrane. In 27 of these wounds the distribution of collagen VII, which represents a specific component of the basement membrane of stratified epithelia, was also analyzed. We were able to demonstrate a virtually identical co-distribution of both collagen IV and VII in the wound area with no significant time-dependent differences in the appearance of both collagen types. Fragments of the epithelial basement membrane could be detected in the wound area from as early as 4 days after wounding and after 8 days a complete restitution of the epithelial basement membrane was observed. In all cases with a wound age of more than 21 days the basement membrane was completely reformed over the former lesional area. The period between 8 and 21 days after wounding was characterized by a wide variability ranging from complete restitution to deposition of basement membrane fragments or total lack of the epidermal basement membrane.


Time-dependent appearance of myofibroblasts in granulation tissue of human skin wounds. Human skin wounds (66) inflicted between 20 h and 7 months prior to biopsy were studied. In order to identify the type of cellular differentiation of the fibroblastic cells in the granulation tissue, alpha-smooth muscle actin and desmin were immunohistochemically localized. The value of any presumed time-dependent appearance and/or disappearance of positively stained cells was tested for the estimation of wound age. In skin specimens with a wound age less than 5 days (n = 15) no typical granulation tissue had developed and no alpha-actin-positive myofibroblasts could be detected. The first appearance of positively reacting myofibroblasts was noted in a 5-day-old wound. In 57% of the lesions with a wound age between 5 and 31 days (25 out of 44 cases) typical granulation tissue formation was present and myofibroblasts with positive reaction for alpha-smooth muscle actin could be identified. Numerous positively reacting cells could generally be found in wounds aged between 16 and 31 days, but also in wounds less than 16 days old. In 29% of the cases with a wound age of more than 31 days (2 out of 7 cases) alpha-sma-positive myofibroblasts also occurred. Fibroblastic cells positive for desmin could not be seen at all in our series. Our results demonstrate the appearance of alpha-sma-positive myofibroblasts with the initial formation of typical granulation tissue in human skin lesions as early as approximately 5 days after wounding. In contrast to recent experimental results these cells remained detectable in wounds aged more than 2 months in some cases.(ABSTRACT TRUNCATED AT 250 WORDS)


Prehospital thrombolysis in a rural community: short- and long-term survival. In order to assess the feasibility and outcome of using prehospital thrombolysis in acute myocardial infarction in a rural community, we performed an open randomized study of patients with symptoms of acute myocardial infarction of less than 6 hours. One hundred and forty-five patients with acute myocardial infarction were allocated to receive IV streptokinase prehospital by means of a mobile coronary care unit (MCCU) (n = 43) or to receive IV streptokinase in hospital (n = 102). The mean delay time to treatment was 138 minutes (MCCU group) and 172 minutes (hospital group) (p less than 0.02). Reperfusion time was 88 minutes for the MCCU group and 92 minutes for the hospital group. Mortality at 14 days was 2.3% for the MCCU group and 11.7% for the hospital group (p less than 0.05). Six month mortality was 4.9% for the MCCU group and 17.3% for the hospital group (p = 0.03). Mortality at 1 year was 6.1% for the MCCU group and 20.0% for the hospital group (p = 0.04). There were no significant adverse events in either treatment group. Thus, prehospital thrombolysis by streptokinase is feasible and allows significant reduction in the delay time to treatment initiation. There are encouraging improvements in both short- and long-term survival with no apparent reduction in safety profile.


Influence of the rt-PA dose (1 mg/kg versus 1.5 mg/kg) and duration of administration on the patency of infarct-related coronary arteries in 81 patients. This open study compared the efficacy and side effects of two dose regimens of intravenous rt-PA initiated before the fifth hour after the onset of myocardial infarction. The first 40 patients (group A) received 1 mg/kg of rt-PA infused over 90 minutes, including a 10-mg initial bolus. The following 41 patients (group B) received 1.5 mg/kg (20-mg initial bolus, 60% over 60 minutes and 40% over 120 minutes). Noninvasive clinical, electrocardiographic, and biochemical parameters of reperfusion were recorded systematically, along with serial measurements of fibrinogen and hemoglobin levels. Coronary arteriography and left ventriculography were performed at the 48th hour or earlier, on an emergency basis, in the absence of signs of reperfusion, or if there were clinical and ECG signs of rethrombosis. Patency of the infarct-related artery (TIMI grades 2 and 3) was achieved in 55% of the group A patients vs. 83% of the group B patients (p less than .01). Twelve emergency coronary arteriographies were performed in each group. No significant difference was observed in the ejection fraction (48% in group A vs. 52% in group B). The peak CPK level was similar in both groups, but the peak occurred earlier in group B (p less than .001). There was no statistically significant difference between the two dose regimens in terms of reduction in fibrinogen or plasminogen levels. Two deaths occurred in group A vs. one death in group B. No cerebrovascular accidents occurred in either group.(ABSTRACT TRUNCATED AT 250 WORDS)


The total ischemic burden European trial (TIBET): design, methodology, and management. The TIBET Study Group. TIBET is a European multicenter, double-blind parallel group trial with the main objective of investigating whether total ischemic burden has important prognostic implications in patients with stable angina on treatment. A secondary objective is to compare the antianginal and antiischemic effects of atenolol (50 mg bid), nifedipine (20-40 mg bid), and their combination using standardized exercise testing and Holter monitoring techniques. The main primary end points are cardiovascular morbidity and mortality. The secondary end points are time to onset of significant ischemia, angina on exercise stress testing, exercise capacity at onset of angina, 1-mm ST-segment depression and termination of exercise, total duration and number of significant ischemic episodes during 48 hours of Holter monitoring, and their circadian distribution. The target population, the assessments, and the management of the trial are described in detail.


Intermittent infusion of dobutamine in the therapy of severe congestive heart failure--long-term effects and lack of tolerance. Twenty patients with refractory heart failure NYHA class IV were randomly assigned to infusion therapy with 9.25 micrograms/kg/min dobutamine over 24 hours or placebo. Eight infusions over a 4-week period were performed in the hospital; between infusions breaks of 3 days were scheduled. A dose titration was performed before study during which dobutamine was infused at 2.5 micrograms/kg/min and increased by 2.5 micrograms/kg/min steps every 15 minutes up to a maximum dosage of 10 micrograms/kg/min. After dobutamine, exercise duration on the treadmill stress test increased from 177 +/- 110 seconds to 251 +/- 120 seconds (p less than 0.05). The heart-rate response to exercise increased (91 +/- 20 to 116 +/- 26 beats/min at baseline, 88 +/- 17 to 132 +/- 26 beats/min after therapy). Body weight decreased from 70.9 +/- 15.5 to 68.9 +/- 14.2 kg (p less than 0.03). On placebo, no significant changes were evident. Systolic time intervals and hemodynamic parameters showed only minor and not significant changes in both groups. No excess mortality emerged during intermittent dobutamine therapy. No clinical or hemodynamic signs of tolerance development were evident during control assessment 3 days after the last infusion. Intermittent therapy with dobutamine seems to be a promising concept in the management of refractory severe heart failure.


Effects of altered extracellular potassium and pacing cycle length on the class III antiarrhythmic actions of dofetilide (UK-68,798) in guinea-pig papillary muscle. The effects of altered extracellular K+ concentrations ([K+]o) and pacing cycle lengths (CLs) on the electrophysiological actions of dofetilide (UK-68,798), a potent class III antiarrhythmic agent, were examined in isolated guinea-pig ventricular papillary muscle. At a normal [K+]o (4 mM) and at CL between 300 and 5000 msec, dofetilide (10 nM) significantly increased the action-potential duration (APD) and the effective refractory period (ERP), whereas other action-potential parameters were unaffected. Elevation of [K+]o to 10 mM reduced membrane diastolic potential (MDP), action-potential amplitude (APA), and the maximum rising velocity of the action-potential upstroke (Vmax). These changes were accompanied by a small shortening of APD90, but with an increase in ERP; i.e., the ERP/APD90 ratio was increased. Dofetilide also significantly lengthened APD90 at 10 mM [K+]o and at each CL. Even at the short cycle lengths (300 and 500 msec), dofetilide-induced increases in APD90 were not attenuated whether [K+]o was at 4 or 10 mM. These results indicate that at various pacing CLs, 10 nM dofetilide increases myocardial APD and ERP to a similar extent without significant reverse use-dependence when the cell membrane is normally polarized or partially depolarized by elevated [K+]o. Dofetilide may, therefore, be expected to be beneficial in the treatment of cardiac tachyarrhythmias related or unrelated to regional myocardial hyperkalemia during myocardial ischemia.


Modified enucleation via lateral orbitotomy for choroidal melanoma with orbital extension: a report of two cases. We report two cases of choroidal melanoma with massive orbital extension that were managed by modified enucleation by way of a lateral orbitotomy. In both cases, the orbital component of the tumor was removed intact along with the globe. This approach allows placement of an orbital implant and avoids the long healing process associated with orbital exenteration.


Measurement of globe position in complex orbital fractures. I. A modification of Hertel's exophthalmometer, using the external auditory canal as a reference point. Where there has been displacement or removal of the lateral orbital rim or zygomaticomaxillary complex secondary to trauma or surgery, the measure of globe position by the Hertel exophthalmometer is problematic and inaccurate. A modification of Hertel's instrument that uses the external auditory canal (EAC) rather than the orbital rim as reference point has been developed. Measurement of globe position of 120 normal volunteers was obtained with the Hertel and the modified EAC-fixated exophthalmometer. Although the mean absolute difference between right and left measurements for both methods (Hertel mean, 0.36 mm; modified mean, 0.51 mm; n = 120) differed statistically from 0 (p = 0.0001), that difference was not clinically significant. A more meaningful comparison was that of the variability of the two methods. The variance of the Hertel method (0.32) did not differ from the variance of the modified method (0.40; p = 0.06). No statistically significant difference was noted for sex (p = 0.33) or for race (p = 0.11). We believe this instrument to be of practical use in the assessment of enophthalmos or exophthalmos in patients whose facial deformity, trauma, surgery, or disease precludes the use of a rim-based exophthalmometry.


Measurement of globe position in complex orbital fractures. II. Patient evaluation utilizing a modified exophthalmometer. Globe position was assessed by both the Hertel exophthalmometer and the modified external auditory canal (EAC)-fixated device in 27 patients with complex orbital fractures (18 tripod and 9 Le Fort fractures). Although 94% of the patients with tripod fractures had relative exophthalmos on the fracture side or no difference between eyes by Hertel exophthalmometry, greater than 30% of the same patients showed relative enophthalmos when measured by the EAC-fixated device. In three of four patients undergoing surgical repair of the orbital floor, modified exophthalmometry showed exophthalmos of greater than or equal to 2 mm postoperatively on the fractured side. The relatively low incidence of enophthalmos in tripod fractures indicates a need for selective orbital floor repair; the uniform exploration of the orbital floor should be discouraged. Multiple comminuted facial fractures (Le Fort II and III) showed a greater variability in globe position and a high frequency (90%) of enophthalmos, suggesting a need for early orbital repair in these patients. EAC-fixated exophthalmometry can provide meaningful information regarding globe position in orbitofacial fracture patients in which orbital rim-based methods are precluded.


Worsening of blepharoptosis in downgaze. We report a group of patients with involutional blepharoptosis most pronounced in downgaze. Retrospective analysis was performed on all patients who underwent involutional blepharoptosis repair in a 30-month period. Physical examination and visual field testing confirmed the increased blepharoptosis in downgaze. The average increase in visual field obstruction is 12.5 degrees and the drop in the upper eyelid relative to the visual axis is 1.9 mm. We conclude that the increased visual field obstruction in downgaze in this subset of patients is significant and deserves clinical evaluation.


Acquired ptosis: dehiscences and disinsertions. Are they real or iatrogenic? Acquired ptosis has previously been felt to be due to a dehiscence or disinsertion of the levator aponeurosis. In the early years of levator surgery, the aponeurosis was often exposed by using blunt dissection. We have found that with a change in surgical technique, which eliminates blunt dissection, the incidence of aponeurotic defects decreases dramatically. Dehiscence and disinsertion may therefore be caused by traumatic surgical dissection of an attenuated aponeurosis.


Complete ptosis as a result of removal of epibulbar lipodermoid. Partial ptosis following epibulbar epidermoid removal is a known complication. The mechanism is generally thought to be symblepharon following aggressive dissection, injury to the levator aponeurosis, or injury to the branch of the ocular motor nerve to the levator muscle. We now report a case of complete ptosis thought to be neurogenic in origin. The possible mechanism is discussed.


Botulinum A toxin for (expressionistic) ptosis overcorrection after frontalis sling. Botulinum A toxin was injected into the frontalis muscle in two patients with complete third nerve palsies to limit intermittent upper lid retraction after a frontalis sling procedure. This form of lid retraction is noted during periods of active facial movement with occipitofrontalis muscle contraction. Although upper lid position may be symmetric when the facial muscles are adynamic, the upper lid may retract during periods of active facial expression. This type of lid retraction was corrected using Botulinum A toxin injections into the frontalis muscles, without affecting the lid position when the facial muscles are adynamic. Both improvement in appearance and intermittent exposure were noted in both cases. Additionally, a blunting of the transverse forehead creases occurred over a defined area after this injection, representing a clinical example of a denervation field produced by a point injection of botulinum toxin.


Blastomycosis of the eyelid and conjunctiva. A 37-year-old man with a recurrent papillomatous lesion of the upper eyelid and four separate bulbar conjunctival ulcers was found to have North American blastomycosis. This represents the first reported case with conjunctival lesions that were not simply contiguous with eyelid involvement. This case underscores the importance of considering blastomycosis in the differential diagnosis of granulomatous conjunctivitis and when examining a lesion of the eyelid resembling a squamous cell carcinoma or atypical papilloma.


Spontaneous orbital subperiosteal hematomas in patients with liver disease. Spontaneous orbital subperiosteal hematomas are rare entities. They are often associated with systemic coagulopathies. We present two cases of spontaneous orbital subperiosteal hematomas in patients with systemic coagulopathies resulting from liver disease. One patient was diagnosed with hepatitis B and had spontaneous resolution of the hematoma. Another patient had alcoholic cirrhosis and required an orbitotomy for drainage of a hematoma for compressive optic neuropathy. With the rising incidence of hepatitis B and the prevalence of alcoholic liver disease, it is important to include liver disease in the systemic evaluation of patients with spontaneous orbital subperiosteal hematomas.


DNA flow cytometry of sebaceous cell carcinomas of the ocular adnexa: introduction to the technique in the evaluation of periocular tumors. DNA content abnormalities are well recognized in tumor cell biology and kinetics. We introduce the technique of DNA flow cytometry through the study of the sebaceous cell carcinomas of the ocular adnexa. By correlating the data to the standard histopathologic parameters of pagetoid changes, degree of anaplasia, and stromal inflammation, significant associations are revealed. All aneuploid tumors demonstrate pagetoid spread, and more severe anaplasia and stromal inflammation. All diploid tumors are nonpagetoid and have lesser degrees of anaplasia and stromal inflammation. A complete review of the technique with a discussion of the implications and applicability to the study of ocular adnexal tumors is presented.


Complications of surgery for orbital tumors. Orbital tumors represent a small but significant portion of ophthalmologic surgery, and few reports of the associated complications are available. This retrospective review was undertaken to assess the overall complication rate of a large series of orbital tumors, as well as to determine possible risk factors that may guide surgeons in the future. There were 137 orbital tumors managed by a total of 14 surgeons at one hospital during a 7-year period. There was an overall 12.4% complication rate (17/137), affecting only 3 of 97 anterior orbitotomies (3%), but 14 of 40 lateral orbitotomies (35%). The location appeared to be the most significant factor, with the majority of complications associated with intraconal tumors.


[Clinico-statistical observations on malignant ocular tumors surgically resolved at the Cluj-Napoca Ophthalmological Clinic between 1970 and 1990]. During the period mentioned above there ware hospitalized 54 patients having epibulbar tumours. The anatomopathological exam which was effectuated for 33 patients hes shown Bowen's disease for 14 cases, spinocellular carcinoma for 15 patients and basocellular for 3 patients. There were 4 cases for epibulbar precancerous. There was retinoblastoma for 19 cases; it had been practiced the enucleation of ocular globe for 17 patients and for two cases it had been practiced the exenteration of the eye socket. At the same period there were 91 uveal melanosis 6 of which having iridial localization, 10 cases of ciliary body [correction of cylar corpus], 75 cases of choroid. The hepatic scintigraphy was practiced for 56 patients and had shown for 8 cases hepatic metastasis. The histopathological exam of the melanomata of ciliary and choroidal corpus had shown type A fusiform cells for 22 cases, type B for 43 cases, epithelioid cells for 6 cases and mixed type cells for 13 cases. The hepatic metastasis were epithelioid cells for all the cases and, for 2 cases there were with mixed type cells, It is necessary an active discovering of intraocular and episcleral malign tumours, for increasing the chance of an earlier diagnosis during a stadium that would be able to be treated as conservator as possible be.


[Open-angle glaucoma yesterday and today]. Glaucoma with open angle is now considered as an early optical neuropathy, that is determined by an uneasiness in flowing of the aqueous humour at the trabecular level. The ocular hypertension is the most frequent manifestation of the evolution of glaucoma, but it is not obligatory because there is a glaucoma with open angle that develop with low pressure. The automatically perimetry and the photography of the retinal arcuate fibers in aneritre light, may evidence incipient lesions before increasing of the ocular tension. Stereophotographies of the optic papillae make possible the calculus of the neuroretinal ring surface that replaces the old ratio cup/disk. The investigation of the microcirculation of the papillae, the determination of the papillar blood tide by pneumotonography and the velocimetry laser Doppler have brought new data for determining the incipient lesions of the optic disk. The large utilization of beta-blockings has changed the life of a glaucoma patient and the filtrant surgery has extended and diversified so being prolonged the visual function.


[Retinal venous obliteration and general pathology]. The study of 148 retinal venous obliterations have shown 81 occlusions of central vein and 67 of I and II venous branch. A number of 90 was for the feminine gender (sex) and 59 for the masculine sex. The average age for the appearance of the venous occlusions was 62 years old, having extreme limits between 36-84 years old. Bilaterality has been for 3 cases. Concerning the associated medical affections, hypertension was for 67 patients, myocardiosclerosis have been mentioned for 67 patients, atherosclerosis for 21 patients, pulmonary scleroemphisis for 12 patients. Arterial hypertension with its aspersion that is arteriosclerosis are the main factors that have generated retinal circulation modifyings and have led to a degree of arterial insufficiency. Comparing the ophthalmological aspect to the pressure in the ophthalmic artery, most of the patients had a concordance of TACR and the retinal and choroidal angiosclerosis. The oscillometric examination to the inferior members has been effectuated for 21 patients and it has shown diminished values only for 3 cases. The forecast of the disease is still reserved. Following a group of 40 patients having OVR between 5 and 15 years old it has been established an average survival of 6.2 years. It is mentioned that 26% between these have dyed during the first six years.


[The eye manifestations in phosphorus-calcium metabolic disorders]. The investigation of a number of 286 patients having disorders of binocular seeing shows a very frequent association of these to spasmophilia, especially for persons of feminine sex during their third or fourth decades of life. The medicamentous treatment of spasmophilia has ameliorated the disturbing binocular seeings, which had appeared again after stopping the treatment for a long time. The authors recommend the cover-test end the examination using Maddox's experiment for tracing rapidly out the binocular seeing disturbings. In order to stimulate the movements of the ocular globe in an reverse sense to the deviation formed by the oculomotor existed lack of poise, it had been used the prescription of classes with prisms assembled conversely to the classic rules, having the indication of wearing them for 3-4 hours a day.


[The serology of toxoplasmosis in uveitis]. Determinations for decelerating the antitoxoplasma anticorps that have been effectuated for 696 uveas proved positivity at 123 aerums (17.6%). The repartition of the positive serology concerning the clinic form of the uvea has proved 48 iridocyclitis, 26 serous central corioretinals, 23 in panuveitis, 13 in posterior uveitis, 11 muscular chorioretinitis, 2 in hyalitis. Taking into consideration only the equal or less than 1:160 titrures, these have been at 7 iridocyclitis, 4 serous central chorioretinitis, 4 panuveitis, 3 posterior uveitis and 3 atrophic central chorioretinitis. It is shown that the diagnosis of ocular toxoplasma must be effectuated just corroborating the clinic data with the positive serology. The antiparasitic treatment doesn't influence upon the low titrures of serous anticorps, which generally maintain at the same value all the life.


[Heredity in myopia].

[Heredity in myopia]. The authors recall various theories that try to explain the appearance and the evolution of the short-sightedness. It is noticed the fact that none of these theories can include all the cases, but heredity proves a variable percentage. The examination of 200 cases of short-sightedness shows for 120 cases an hereditary factor. The transmission of it was dominant with variable penetration. Concerning the short-sightedness as a disease or malign one, the authors talk about a genetic condemnation of the sclera.


[The marking of the anterior crystalloid in the extracapsular extraction of the crystalline lens by the intrasaccular (in the bag) technic]. The authors try to mark the anterior crystalloid before sectioning it, using the chemical touch (solution three coloured acetic or sodium hydroxide) or by means of physical methods (electrocoagulation or diathermy for minimum electric power). In vivo the two last methods have given stable whitish points by the middle of which had been executed the sectioning of the crystalloid. The authors emphasize the advantages that are known by the technique of the extracapsular extraction of the crystalline in the bag and the simplifying of the manoeuvre for marking the anterior crystalloid.


[Obesity as a risk factor in diabetic retinopathy]. The relations between the glucidic and lipidic metabolism fatness and diabetes are known for a long time. The fatness as a factor of risk for the diabetic retinopathy and consecutively for the appearance of the cecity have less been studied. There are studied the variations of lipids, cholesterol, the fatness (for 3 degrees by Broca) for a group of diabetic patients, type I or II, the age of the patients (between 18-80 years old) there are presented the retinal modifying: arterial, venous, microaneurysm, haemorrhages, the macular modifying. In comparison to the normoponderal diabetic patients, for these who are fat patients especially I or II degree, all the modifying are exaggerated. There was chosen the fatness from the lots of the factors of diabetes'risk, because I think it may be influenced. The treatments of retinopathy are few and less encourage. The doxium and the photocoagulation could eventually make the cecity to be late but they belong to the group of treatment, but it is not a prophylactic element for cecity. The real prophylaxis has to begin since the childhood with a normocaloric food diet. The prophylaxis of the diabetes is the true prophylaxis of the cecity.


[Spontaneous luxation of the crystalline lens into the anterior chamber (a clinical case report)]. A patient of 45 years old presents a sprained crystal in the anterior chamber, with secondary acute glaucoma at the left eye and surgically aphakia with ciliary staphyloma and optic atrophia for the right eye. VOR zero; VOL-PL; TOR-17,3 mm Hg; TOL-81, 9 mm Hg. After hypotension of the eye using a perfusion of mannitol and acetazolamide the ocular tension is normalized. It is practiced the extraction of the lens from the anterior chamber with a good post-surgically evolution. The disease may be joined to Marfan, Marchesani, syndrome homocystinuria or an early diagnosis of the zone without any manifest cause.


[Toxoplasmic chorioretinitis and hydrocephalus]. A child of 4 years old presents cicatricial toxoplasmic chorioretinitis, hydrocephaly and neurological disturbance. The titre of serous anticorps antitoxoplasma have been presented at 1/10. The child doesn't fix the objects, he presents nystagmus of fixation. Being in the presence of a case of congenital toxoplasmosis sequela stadium, it is not necessary a treatment. We insist upon a periodically control for the pregnant women and the treatment for those having positive serology. There are also necessary the prophylactic orders respecting the norms of hygiene and especially the use of milk and meat after an adequate thermic treatment of it.


[The etiopathology of phacoantigenic uveitis and phacolytic glaucoma]. The phacoantigenic uveitis is a self-immune affection at the lens proteins, which may be determined by a 3rd-type hypersensitivity, and some sterner forms by a 4th-type hypersensitivity. In phacolytic glaucomas, the macrophagic reaction at degraded lens proteins obstructs the trabecular gaps, determining the raise of ocular tension with the appearance of secondary glaucomas.


[Correction with contact lenses. The components of correction with contract lenses]. In order to understand the way contact lens adapts, we considered as a necessity the successive analysis of the components participating of the correction. Thus we have presented the role of the cornea and the evolution of the tear film under the contact lens. Afterwards contact lens of different types, corneo-scleral, hard or soft as well as the discontinuous, prolonged and permanent wear of lens have been presented.


[The staging of malignant uveal melanoma]. There have been investigated 30 malignant uveal melanomas hospitalized in the Ophthalmologic Clinic of Craiova during the last 15 years. The disease has developed more frequently among the female sex in a 3/2 proportion, the maximum age ranging between, 40 and 70 years old. As to the localization, 2 were at the level of the iris, 7 at the level of ciliary body and 21 at the choroid level. In anterior localizations tumour extirpation was possible in the 2 cases through iridectomy and in only one case through cyclectomy. Anatomicopathologically, most cases presented mixed-type cells where prevailed the type B fusiform cells and epithelioid cells with big quantitative variations from one zone to another. The tumours detected in an incipient stage benefited from laser photocoagulation or application of CO60 and I125 radioactive disks. In advanced stages enucleation was practised preceded by 2 treatments of radiotherapy for the devitalization of tumorous cells and prevention of metastases.


[The genetic predisposition in open-angle glaucoma]. The genealogic tree examination of a female patient with open angle glaucoma showed the presence of eight more cases of glaucoma in the same family. The transmission was autosomal predominant with a 75% penetration of the pathological gene. In the patient's family the affection appeared practically at the same age and evolved with the same specific feature.


[The echographic diagnosis of intraocular tumors in children]. The echographic exam done on 14 children with leukocoria (white pupil) confirmed in 7 cases the retinoblastoma diagnosis. The exam differentiates the solid tumours from the cystic ones and points out the tumours with non-typical clinical aspect. Results from here the great importance of echography effectuation, especially in non-typical forms with difficult clinical diagnosis. The association of the echographic examination, together with the classical methods of investigation, contributes to the establishing of a correct diagnosis of the size and exact extension of the tumour.


[The importance of the pleoptophor in treating children with amblyopia]. The author presents her own experience in pleoptic treatment, using the occlusion and the exercises at the amblyophor, on 161 children, during a 12 years period of time. The results of the therapy lead to cure in 38% of cases, to improvement in 46% of cases and fail in 16% of cases. The indications, the preparation, the practical development of the treatment at pleoptophor are discussed; the reasons of the failures are carefully analyzed. It is emphasized that the good results are due to the tracking down of children with strabismus in kindergartens, to the application of the amblyopic preventive optical treatment and to pleoptic treatment maintained for years with monthly checkups to prevent relapses.


[The practical aspects of the monocular wearing of a contact lens]. The contact lens was prescribed to 64 cases, 38 of which having postsurgical monolateral aphakia. The lens extraction was made in 28 cases with traumatic cataract, 7 cases with complicated cataract and 3 cases with monolateral congenital cataract. The application of contact lens was made from two months to three years after the cataract operation; 6 patients wore before the surgical act usual glasses to correct the aphakia. After that the patients were called back monthly to controls; we used these opportunities to collect data about the advantages and disadvantages of contact lens wearing.


[Melanosis oculi and malignant choroid melanoma]. The patient aged 75 years presents since her youth, in the left eye, ocular melanosis with abnormal pigmentation of the sclerotic, hyperpigmentation of the iris and abnormal pigmentation of the fundus oculi. Six months ago, she noticed that she is not able to see with this eye, and the fundus oculi examination emphasizes a malignant choroidal melanoma, with a fixed detachment of the retina. The authors show that the denomination Bourquin's benign ocular melanosis, under which the affection is known, is improper, as 10% of the cases of ocular melanosis degenerate into melanosarcomas. The term melanosis oculi is the most adequate one.


[The artificial crystalline lens implant in diabetic patients]. Are presented the results obtained by artificial lens pupillary implant (Fedorov) at 22 eyes, 18 patients between 48-81 years old, to which the development of cataracta was characterized by a great difference between the eyes. It have been obtained 63% very good visual acuity (5/10 and over) and 31% good (5/15-5/40). The authors Conclusion is the artificial lens implant may be done on diabetic patients if FO doesn't have great modifications.


Morphological characterization of exovesicles and endovesicles released from human erythrocytes following treatment with amphiphiles. In order to morphologically characterize exo- and endovesicles released during treatment of erythrocytes with amphiphiles and to look for possible amphiphile-specific effects on the vesiculation pattern, human erythrocytes were treated at 37 degrees C with amphiphiles at concentrations where they exhibit maximum protection against hypotonic haemolysis (cAHmax). Released exo-and endovesicles and treated cells were studied by means of transmission (TEM) and scanning (SEM) electron microscopy. All sphero-echinocytogenic amphiphiles induced a release of both spherical and tubular exovesicles. Dodecyl maltoside, a nonionic amphiphile with a bulky polar head, induced a release of predominantly tubular exovesicles, while all other sphero-echinocytogenic amphiphiles induced a release of predominantly spherical exovesicles. Some branched tubular exovesicles were released by a double-chained cationic amphiphile. Tail- and tongue-like structures were often seen on the exovesicles. Spherical exovesicles were frequently invaginated. Stomatocytogenic amphiphiles induced endovesiculation. In erythrocytes treated with most of the stomatocytogenic amphiphiles the endovesicles were clustered, but with some amphiphiles the endovesicles were randomly distributed. Large ringformed endovesicles (octaethyleneglycol alkyl ethers) and endovesicles in chains (octyl and decyl glucopyranoside) also occurred. The endovesicle membrane was often budding onto the lumen of the vesicle and in some cases this could ultimately lead to a vesicle inside the endovesicle. We conclude that amphiphiles do not only trigger vesiculation, but may also specifically affect the vesiculation processes.


[A tuberculous etiology in Mikulicz's syndrome]. A patient 20 years old presents bilateral hypertrophy of the lacrimal glands with discreet exophthalmos and conjunctival injection at the upper supero-external fornix level. The ocular affection appeared 6 weeks before as a painless tumefaction initially at the right eye and after that at the left eye also, with palpebral fold reduction. From childhood the patient has a ganglionary tuberculosis confirmed by the biopsy of a lateral cervical ganglion. Under the rim of the orbit on palpate the lacrimal gland as a depressible, painless mass, which is continued in the orbit. A lacrimal gland biopsy, effected at the left eye, shows the presence of dispersed nodules with round cells in the whole fragment of lacrimal gland, and epithelioid and giant cells in the center. The inflammatory phenomenon yield slowly with tuberculostatics generally and hydrocortisone in local instillations.


[The ocular manifestations in craniopharyngioma]. 11-years-old patient, for which the cranial radiograph shows modifications specific to craniopharyngioma, presents diminution of visual acuity at both eyes, psychic retardation and signs of insipidus diabetes. It is emphasized the unusual association between craniopharyngioma and exophthalmos, discordantly appeared on a background of global glandular insufficiency.


Induction of cystine transport in bovine pulmonary artery endothelial cells by sodium arsenite. Sodium arsenite is one of a number of agents reported to induce a 30-34 kDa 'stress' protein in cells. Other agents which induce this stress protein, including diethyl maleate (DEM) and H2O2, have also been reported to be inducers of cystine transport in fibroblasts, macrophages, endothelial cells and other cell types. We have determined that micromolar levels of sodium arsenite increase cystine transport in bovine pulmonary artery endothelial cells (BPAEC), resulting in increases in intracellular glutathione (GSH). The increase in cystine transport appears to be due to stimulation of the synthesis of a protein analogous to the xc- transport system, a sodium-independent system specific for cystine and glutamate. We have determined that this stimulation is maximal between 8-16 h after addition of sodium arsenite and is inhibited by exogenous GSH. Others have reported that synthesis of the 30-34 kDa stress protein is maximal between 2-4 h and returns to baseline by 6-10 h. We conclude that cystine transport may be considered a 'secondary' stress response and is likely to be modulated by sulfhydryl-reactive agents.


Stimulation-enhanced 3-O-methylglucose efflux from the frog sartorius: kinetics and properties of the system. The characteristics of the process by which contraction enhances glucose transport in the frog sartorius were studied. Electrical stimulation increased the permeability of muscles to 3-O-methylglucose (3-O-MeGlc), a nonmetabolizable glucose analogue, increasing efflux as well as uptake. Enhanced efflux was due to an increase in Vmax of the efflux process. A lactacidosis had no effect on basal 3-O-MeGlc efflux, and replacement of media Na+ with Li+ did not affect stimulation-induced uptake. Also, basal and stimulated uptake was not affected by 1 microM 12-O-tetradecanoylphorbol-13-acetate (TPA), a protein kinase C activator. Lastly, N-carbobenzoxy-glycyl-L-phenylalaninamide, which inhibits insulin-enhanced, but not basal, glucose uptake in adipocytes, inhibited both basal and stimulated 3-O-MeGlc fluxes in the frog sartorius. From these findings, we conclude: (1) contraction and exercise enhance glucose transport in muscle by increasing the number of transporters in the plasma membrane, or their turnover, by an unknown process; and (2) basal glucose transport of muscle, unlike that of adipocytes, can not be distinguished from stimulated transport on the basis of its insensitivity to N-carbobenzoxyglycyl-L-phenylalaninamide.


[The invasion of neighboring structures in malignant choroid melanoma]. It has been followed the microscopical evolution of the malignant melanoma of the choroid. After a stage of local growth, often with necrosis and intratumoral haemorrhages, it invades the neighbouring structures. The invasion of retina is produced at the apex level, making a breach in Bruch's membrane, the tumour becomes likewise a dumb-bell, a mushroom or a clepsydra. When the tumour is in contact with the retina, it produces a proliferation of pigmentary tumoral cells, glial cells and lymphoplasmocytary inflammatory reaction, which emphasize the sensorial retina detachment. The sclera invasion takes place especially in plaqued melanoma, through the points of minimal resistance which are the nervous routes, breaking up the scleral structures. The appearance of rudimentary vessels, with tumoural cells in lumen, constitutes a serous prognostic because of tumoural embolisms.


Identification of a broad-specificity nucleoside transporter with affinity for the sugar moiety in Giardia intestinalis trophozoites. A broad-specificity nucleoside transporter has been identified in Giardia intestinalis trophozoites, using a rapid sampling assay to measure influx of [3H]deoxycytidine, [3H]adenosine and [3H]guanosine at 0 degrees C. The influx of each labelled nucleoside was inhibited strongly by all common, naturally-occurring nucleosides but only poorly or not at all by nucleobases, indicating that the transporter recognizes structural features on the furanosyl moiety of ribo- and 2'-deoxyribonucleosides. Both 2'- and 5'-deoxyadenosine were potent inhibitors of influx (greater than 95% inhibition at 2 mM), whereas 3'-deoxyadenosine was significantly less effective (approx. 70% inhibition), and 2',3'-dideoxycytidine and cytosine arabinoside were virtually inactive (0-20% inhibition). The data reveal that the 2'- and 5'-hydroxyl groups are not necessary for the recognition of nucleosides by this transporter. However, the 3'-hydroxyl appears to be important. Michaelis-Menten constants (Km) were calculated for the influx at 0 degrees C of deoxycytidine (220 +/- 116 microM) and adenosine (45 +/- 24 microM), with respective Vmax values of 13 +/- 4 and 11 +/- 2 pmol min-1 (10(6) cells)-1. Only 12-26% of [3H]thymidine influx occurred through this transporter, the remainder entering the cells through a thymine/uracil-specific transporter described previously. Thymidine exhibited a Ki of 205 +/- 90 microM against [3H]deoxycytidine influx.


[The adaptation to contact lenses].

[The adaptation to contact lenses]. The indications of contact lens adaptation in optical purpose are widely large, starting, with refractive errors [correction of vices], unilateral and bilateral aphakia, myopia, anisometropia and astigmatism, together with the use of contact lens in esthetic purpose. We have been presented the adaptation techniques, the supervise, and maintenance of contact lens.


A low-affinity nucleobase transporter in the protozoan parasite Giardia intestinalis. A membrane transporter with general affinity for purine and pyrimidine bases has been identified in Giardia intestinalis trophozoites by measuring cellular influx of [3H]adenine, [3H]guanine and [3H]thymine at 0 degrees C. The base transporter is distinct from the thymine/uracil-specific (type 1) and broad-specificity (type 2) nucleoside transporters of G. intestinalis. Influx of each labelled base was retarded by unlabelled bases, with inhibition in the order: hypoxanthine greater than adenine greater than thymine greater than uracil. The IC50 values for these bases (measured for [3H]adenine influx) were 0.46 mM, 1.15 mM, 1.52 mM and 2.28 mM, respectively. Nucleosides did not inhibit base influx (less than or equal to 15% inhibition at 2 mM, a 400-fold molar excess, at which concentration [3H]nucleoside influx was inhibited by greater than 95%). The Michaelis-Menten constant (Km), calculated for adenine and thymine influx at 0 degrees C, was 1.44 +/- 0.08 mM and 1.61 +/- 0.37 mM, respectively, with corresponding Vmax of 383 +/- 16 and 498 +/- 112 pmol min-1 (10(6) cells)-1. The data demonstrate the existence of a low-affinity, facilitative base transporter with no detectable affinity for nucleosides. The inability of uridine or thymidine to significantly reduce the rate of thymine influx indicates that the previously described thymine/uracil-specific (type 1) thymidine transporter cannot transport thymine, despite its affinity for the base.


The effects of 1,25-dihydroxyvitamin D-3 deficiency on Ca(2+)-transport and Ca(2+)-uptake into brush-border membrane vesicles from pig small intestine. BBMV were prepared from duodenal segments of untreated, 1,25-(OH)2D3- or vitamin D-3-treated rachitic piglets and from non-rachitic controls by the Mg2+ precipitation method. The rachitic piglets were offspring from the 'Hannover Pig Strain' which suffer from pseudo vitamin D-deficiency rickets, type I (no renal 1-hydroxylase activity). Initial uptake of Ca2+ (up to 35 s) at low [Ca2+] (between 0.02-0.25 mmol/l) into isolated BBMV consisted of a saturable and non-saturable component. The apparent Vmax of the saturable component was significantly lower in rachitic piglets than in control piglets. In the presence of an inside negative potassium diffusion potential, the difference in uptake extended over at least 15 min. The apparent Km of Ca(2+)-uptake was not influenced by the rachitic condition. Treatment of rachitic piglets with sequential doses of 1,25-(OH)2D3 for three days (1 microgram/day) or with a single dose (2.5 mg) of vitamin D-3 elevated the saturable Ca(2+)-uptake component to values similar to those of control piglets. Addition of 1 mmol/l verapamil to the vesicular suspension inhibited Ca(2+)-uptake in BBMV of control piglets by 40-60% but was without effect on preparations from rachitic piglets. It was concluded from the study that 1,25-(OH)2D3 dependent Ca(2+)-uptake into BBMV constituted a saturable process which can be inhibited by a known Ca(2+)-channel blocking agent. It appears that 1,25-(OH)2D3 increases the number of verapamil sensitive Ca(2+)-transport components in brush-border membranes. The vitamin D-dependent changes in vesicular Ca(2+)-uptake were paralleled by the expression of an active transmural Ca(2+)-transport across the mucosa.


Opioid peptide interactions with lipid bilayer membranes. The interaction of the delta-opioid receptor selective peptides, cyclic [D-Pen2, D-Pen5]-enkephalin [DPDPE] and its acyclic analog, DPDPE(SH)2, with neutral phospholipid bilayer membranes was examined by permeability and calorimetry measurements. The permeabilities were accomplished by entrapping either peptide inside of unilamellar liposomes (composed of a mixture of a molar ratio 65:25:10 phosphatidylcholine/phosphatidylethanolamine/cholesterol) then monitoring the peptide efflux through the bilayer. The initial permeability of DPDPE (first 12 h) averaged over four experiments was (0.91 +/- 0.47).10(-12) cm s-1. In contrast the average permeability of the acylic DPDPE(SH)2 was (4.26 +/- 0.23).10(-12) cm s-1. The effect of these peptides on the phase transition, Tm, of 1,2-dipalmitoylphosphatidylcholine (DPPC) bilayers was examined by high sensitivity differential scanning calorimetry. The Tm, the calorimetric enthalpy, and the van 't Hoff enthalpy of DPPC were not significantly altered by the presence of DPDPE, whereas the calorimetric data for DPPC with DPDPE(SH)2 showed a small, yet significant, increase (0.2 degrees C) in the Tm with a 30% decrease in the cooperative unit. Both the permeability and calorimetry data reveal a stronger peptide-membrane interaction in the case of the more flexible acyclic peptide.


In vitro cytotoxicity of liposome-encapsulated doxorubicin: dependence on liposome composition and drug release. We have investigated the in vitro cytotoxicity of free doxorubicin (DOX) and liposome-entrapped DOX (L-DOX) against a human ovarian carcinoma cell line (OV-1063) using a colorimetric assay. DOX was encapsulated in the inner water phase of liposomes by an ammonium sulfate-generated proton gradient. Liposomes varied in phospholipid composition but were of a similar size. It was found that the cytotoxic activity of L-DOX is substantially decreased when liposomes containing phospholipids of high phase-transition temperature (Tm) are used. The type of negatively charged headgroup did not have any significant influence on the cytotoxicity observed. Experiments using resin beads that bind free and protein-bound DOX, but do not interact with L-DOX, indicated that the cytotoxic effect is mediated by the release of drug from the liposomes into the extracellular medium; no evidence was found for direct cellular uptake of liposome-encapsulated drug. The use of the ionophore nigericin to induce the release of DOX from high-Tm liposomes increased cytotoxicity to a level comparable to free DOX, suggesting that 'remote release' techniques may substantially improve the efficiency of liposome-mediated drug delivery and allow for the full exploitation of the favorable pharmacokinetic properties of specific high-Tm formulations.


GM1b and GM1b-GalNAc: major gangliosides of murine-derived macrophage-like WEHI-3 cells. WEHI-3 cells, derived from a BALB/c mouse, are a myelomonocytic leukemic cell line with macrophage-like properties. We have isolated, purified and characterized the monosialogangliosides from WEHI-3 cells by 1D-HPTLC, 2D-HPTLC, enzymatic degradation, HPTLC-immunostaining, gas-liquid chromatography and fast atom bombardment-mass spectrometry (FAB-MS). Quantitative 2D-HPTLC shows two monosialogangliosides are the major components, constituting 77% of the total, with a third monosialoganglioside being 3%. The two major components were identified as (NeuAc)GM1b and (NeuAc)GM1b-GalNAc and the minor component as (NeuAc)GM1b-GalNAc-Gal. The presence of GM1b in this myelomonocytic cell line is consistent with its presence in other murine immune cells and tissues. GM1b-GalNAc and GM1b-GalNAc-Gal have been reported in T-lineage cells but not in resident or stimulated murine macrophages. Each of these monosialogangliosides belongs to the asialoGM1 synthetic pathway. Preliminary results indicate a disialo member of this pathway, GDlc, may also be present as a minor component. This ganglioside pathway, containing species which are not sialylated on the internal galactose, appears to be dominant in and may be characteristic of murine immune cells.


Operation and energy dependence of the reducing-equivalent shuttles during lactate metabolism by isolated hepatocytes. The participation and energy dependence of the malate-aspartate shuttle in transporting reducing equivalents generated from cytoplasmic lactate oxidation was studied in isolated hepatocytes of fasted rats. Both lactate removal and glucose synthesis were inhibited by butylmalonate, aminooxyacetate or cycloserine confirming the involvement of malate and aspartate in the transfer of reducing equivalents from the cytoplasm to mitochondria. In the presence of ammonium ions the inhibition of lactate utilization by butylmalonate was considerably reduced, yet the transfer of reducing equivalents into the mitochondria was unaffected, indicating a substantially lesser role for butylmalonate-sensitive malate transport in reducing-equivalent transfer when ammonium ions were present. Ammonium ions had no stimulatory effect on uptake of sorbitol, a substrate whose oxidation principally involves the alpha-glycerophosphate shuttle. The role of cellular energy status (reflected in the mitochondrial membrane electrical potential (delta psi) and redox state), in lactate oxidation and operation of the malate-aspartate shuttle, was studied using a graded concentration range of valinomycin (0-100 nM). Lactate oxidation was strongly inhibited when delta psi fell from 130 to 105 mV whereas O2 consumption and pyruvate removal were only minimally affected over the valinomycin range, suggesting that the oxidation of lactate to pyruvate is an energy-dependent step of lactate metabolism. Our results confirm that the operation of the malate-aspartate shuttle is energy-dependent, driven by delta psi. In the presence of added ammonium ions the removal of lactate was much less impaired by valinomycin, suggesting an energy-independent utilization of lactate under these conditions. The oxidizing effect of ammonium ions on the mitochondrial matrix apparently alleviates the need for energy input for the transfer of reducing equivalents between the cytoplasm and mitochondria. It is concluded that, in the presence of ammonium ions, the transport of lactate hydrogen to the mitochondria is accomplished by malate transfer that is not linked to the electrogenic transport of glutamate across the inner membrane, and, hence, is clearly distinct from the butylmalonate-sensitive, energy-dependent, malate-aspartate shuttle.


Growth of human cultured cells exposed to a non-homogeneous static magnetic field generated by Sm-Co magnets. A static magnetic field, with a strong spatial gradient, was established on the surface of cell culture dishes by use of a gilded iron needle set vertically above an Sm-Co magnet. The calculated magnetic flux density was more than 1.5 T at the center of the needle tip, and the products of the flux density and its gradient were about 200 and 60 T2/m at distances of 0.1 and 0.3 mm, respectively, from the center. The DNA content, DNA synthesis and labeling index of cultured cells located within 0.1 mm from the center of the needle, and the growth rate of cells located within 0.3 mm from the center, were measured. HeLa cells grew at a normal rate for 96 h in the magnetic field and showed no significant change in shape, detectable by scanning electron microscopy. The growth of HeLa cells was not influenced by exposure to the magnetic field. Similarly, exposure for 48 h to the magnetic field had no effect on growth of normal human gingival fibroblasts (Gin-1). The DNA content, assayed by microfluorometry of the nuclei of both types of cells stained by the Feulgen reaction, was not significantly different from that of controls. Moreover, exposure to the magnetic field had no effect on DNA synthesis or the labeling index of HeLa cells assayed by autoradiography of incorporated [3H]thymidine. It is concluded that a non-homogeneous magnetic field of the intensity and the gradient used in this study does not significantly influence the growth of HeLa cells or Gin-1 cells.


Mechanism for endothelial cell injury induced by 15-hydroperoxyeicosatetraenoic acid, an arachidonate lipoxygenase product. The mechanisms for endothelial cell injury induced by the lipid hydroperoxide 15-hydroperoxyeicosatetraenoic acid (15-HPETE), an arachidonate lipoxygenase product, were explored in cultured bovine endothelial cells. In serum-free medium, there was significant incorporation of [3H]-15-HPETE into the phospholipids of endothelial monolayers, and 15-HPETE induced severe endothelial cell injury, which was determined by the 51Cr-release assay. In contrast, in serum containing medium, there was little incorporation of [3H]-15-HPETE into the cells, and no cellular injury occurred. In the serum free condition, [3H]-15-HPETE was mainly incorporated into the phospholipids. The incorporated 15-HPETE produced lipid peroxidation, which was determined by the accumulation of malondialdehyde in the cells. The 15-HPETE-induced lipid peroxidation was suppressed by radical scavengers (MK-447, MCI-186), anti-oxidants (alpha-tocopherol, butylated hydroxytoluene) and iron chelators (desferrioxamine,2,2'-bipyridine). Furthermore, these agents also suppressed the 15-HPETE-induced cytotoxicity. These results indicate that 15-HPETE-induced endothelial cell injury depends on iron-mediated lipid peroxidation.


Mechanisms for plasma-mediated activation of human blood cell aldehyde dehydrogenase. Aldehyde dehydrogenase (ALDH; EC 1.2.1.3) activity assays were carried out on isolated human blood cells in phosphate-buffered saline (PBS) and in PBS mixed with human plasma. In assays with intact erythrocytes or sonicated leukocytes, the presence of 50% (v/v) or greater of plasma in the reaction mixtures produced a 2-fold increase in the rate of aldehyde oxidation. In corresponding assays with sonicated erythrocyte samples, the ALDH activity was enhanced on an average 1.5-fold, whereas a slight decrease was observed in assays with intact leukocytes. The ALDH inhibitor disulfiram almost completely abolished the enzyme activity both in the absence and presence of plasma. In assays with sonicated leukocytes, the activation effect could be antagonized by EDTA, indicating that it was caused largely by divalent cations. With sonicated erythrocytes, a significantly reduced ALDH activity was found only with the highest concentration of EDTA tested, and since a similar reduction was obtained also when plasma was omitted, the plasma-mediated activation of erythrocyte ALDH was suggested to be due to a different mechanism. After separation of plasma by gel filtration, an active fraction was identified by GC-MS and 1H-NMR to contain pyruvic acid, lactic acid and glucose. When tested at physiological plasma concentrations, pyruvic acid caused an increase in erythrocyte ALDH activity similar to that obtained with plasma, while lactic acid and glucose did not. Pyruvic acid did not activate the leukocyte ALDH. Based on these results, it is indicated that the plasma-mediated activation of erythrocyte ALDH is due to pyruvic acid, which reoxidizes NADH via lactate dehydrogenase (EC 1.1.1.27) and, thereby, increases the rate of dissociation of NADH from the terminal enzyme-NADH complex, the rate-limiting step in the ALDH pathway.


Guanine nucleotide regulatory protein levels and function in spontaneously hypertensive rat vascular smooth-muscle cells. We compared G-protein levels and function in membranes from vascular smooth-muscle cells (VSMC) derived from mesenteric arteries from SHR, WKY and Wistar rats. Basal adenylyl cyclase activity was significantly reduced in SHR membranes compared with Wistar, but was similar to WKY. Isoproterenol stimulation (10(-4) M) was significantly lower in SHR membranes compared to WKY, but was similar to that in Wistar, which was also significantly lower than WKY. Forskolin (10(-4) M) and NaF (10(-2) M), resulted in a higher stimulatory response in SHR membranes. Biphasic effects of GTP on isoproterenol-stimulated membranes demonstrated unaltered Gi function in SHR membranes. No significant differences were seen in the levels of Gs alpha (44- and 42-kDa forms), Gi2 alpha and the beta-subunit in immunoblotting studies of the membranes. Amounts of Gq alpha/G11 alpha and Gi3 alpha were also unchanged. In conclusion, there are differences in adenylyl cyclase responses in SHR VSMC membranes which are not a consequence of altered levels of G-proteins, but may reflect genetic differences rather than effects of hypertension.


A human osteoblastic cell line, MG-63, produces two molecular types of macrophage-colony-stimulating factor. A human osteoblastic cell line, MG-63, mouse primary osteoblasts, and a mouse osteoblastic cell line, MC3T3-E1, were shown to produce macrophage-colony-stimulating factor (M-CSF) by bone-marrow-cell colony assay, using a specific neutralizing antibody for M-CSF. Immunoblot analysis of M-CSF, produced by MG-63 cells, revealed the presence of a higher-molecular-weight species of M-CSF, in addition to the 85-kDa M-CSF. The higher-molecular-weight species had a high affinity to the DEAE-Sephacel column and was sensitive to chondroitinase ABC and AC. These physico-chemical profiles were wholly compatible with those of the proteoglycan form of M-CSF (PG-M-CSF), which was recently identified by our group in the conditioned medium of Chinese hamster ovary cells transfected with the 4.0-kb cDNA of the M-CSF gene. Conditioned medium of MG-63 cells was fractionated by DEAE-Sephacel column chromatography, and the M-CSF of each fraction was measured by both enzyme-linked immunosorbent assay and bone-marrow-cell colony assay. The fractions eluted by 0.3-0.6 M NaCl, which were shown to contain only PG-M-CSF on immunoblot analysis, also have macrophage-colony-stimulating activity.


Angiotensin II and active phorbol esters induce proto-oncogene expression in isolated rat hepatocytes. In isolated rat hepatocytes PMA, angiotensin II and to a lesser extent other hormones induce an early genetic response (increased expression of c-fos, c-mos, c-myc and beta-actin) without altering the expression of the glyceraldehyde 3-phosphate dehydrogenase gene. PMA, PDB and O-met-PMA, but not alpha-phorbol, stimulated c-fos expression. The effect of angiotensin II was inhibited by the AT1 antagonist, Losartan (DuP 753) (Ki approx. 25 nM), but not by the AT2 antagonist PD123177. Angiotensin II was much more effective than vasopressin or epinephrine in inducing proto-oncogene expression which suggests that angiotensin II receptors may exert actions in addition to those shared with the receptors for the other calcium-mobilizing hormones. The effect of PMA and angiotensin II on c-fos expression took place rapidly, with half times of 7 and 12 min, respectively. Actinomycin D markedly diminished basal c-fos expression whereas cycloheximide had the opposite effect. Actinomycin D diminished the effect of PMA and angiotensin II but it did not block them. PMA and the calcium-mobilizing hormones increased c-fos expression above the level observed with cycloheximide alone. These data suggest that PMA and the calcium-mobilizing hormones increased both transcription of the c-fos gene and stabilization of the proto-oncogene mRNA.


Gangliosides inhibit platelet-derived growth-factor-stimulated increases in intracellular calcium in Swiss 3T3 cells. Individual Swiss 3T3 cells stimulated by platelet-derived growth factor delivered by means of a picopump device respond with a brisk, large, and sustained increase in intracellular calcium concentration ([Ca2+]i). Preincubation of cells with either GM1 or GT1b gangliosides inhibited the proportion of responding cells and caused a dose-related diminution in the magnitude of the increase in [Ca2+]i. This effect of ganglioside is probably part of the mechanism through which gangliosides exert their biological effects, including inhibition of platelet-derived growth-factor-induced mitogenesis.


[Seasonal changes in the human constant electrical field in various age groups]. Seasonal changes of the constant electric field of healthy people of various age groups were studied. The constant electric field was shown to be characterized by a fine structure of distribution of electric potential differences (EPD) along the skin in relation to a referent point located on the neck in the intersection region of trapeziform and clavicular-nipple muscles. The constant electric field of the people of all ages undergoes seasonal changes involving displacement of all EPD values to the positive region during autumn-winter period as compared to the spring-summer one. It is suggested that such EPD change is conditioned by a change of the organism metabolic activity. The discovered differences in EPD displacement values of various age groups can serve as physiological characteristics of transitional processes in the growing organisms.


[Origin and evolution of life and intellect from the point of view of information processing]. Beginning of life and intellect on the Earth is determined by the fact that synthesis of semantic information about algorithms of chemical transformation for appointed classes of chemical compounds is not accompanied by the information synthesis, but is controlled by chance in genetic material. Genetic information as memorizing accidental choice always arises as a result of the environment interaction, when memorizing realization in the form of repeated reproduction occurs on the basis of processes radically different from those which created initial genetic chance. Such stepped synthesis of information has hierarchical character and it is described by entropy of different dimensions. Entropy is determined by conditional probabilities. Therefore as a form of life becomes complicated, the entropy is decreased within the limits of the given hierarchical step. The part of entropy depending on the number of elements of the system (cells, individuals, etc.) always increases in the memorizing process, i. e. life defines the maximum of entropy production (but not the minimum like in nature). This fact settles paradoxes of chance in Darwinism: as a result of entropy decrease within higher hierarchical step the simbiotic processes for more complex forms of life can arise with great probability. For instance, rapidity of evolution of human brain given by supplementary chance in the mechanism of fecundation under the conditions of biochemical pressure of androgenes both, on the brain development and on muscular force at the same time. Synthesis of information in the brain is determined by the same principles, but extremums of the thermo-dynamic potential (their analogues in logic) are based on an arbitrary system of axioms. As a whole life and intellect are not fluctuations pointed against entropy increases, but they arose according to spontaneous process of entropy development.


[Activation of superoxide dismutase molecules under the effect of ultraviolet radiation]. The effect of UV light (240-390 nm) with dose range 1.51-30.2 x 10(2) J/m2 on the functional properties of superoxide dismutase was investigated at various pH values. Photoactivation was shown to be maximum at pH values extreme for the display of superoxide dismutase activity.


[Change in adaptability and kinetics of T-lymphocyte adaptation to hypotonia in pathological states of mammals]. The kinetics of adaptation of the volume of T-lymphocytes and their membrane potential in a hypotonic medium is investigated. It is shown to have a quasiperiodic character. The shortest adaptation period T = 2 min characterizes the adaptation of T-lymphocytes test animals, the most complete adaptation of the volume and the membrane potential in hypotonic medium. Upon adaptation their values are only 6% higher than respective values in an isotonic medium. If an aseptic inflammatory process develops in a mammalian organism, the adaptation period increases to T = 2.5 min, while the volume and the membrane potential remains 28% higher than in an isotonic medium. In the course of development of induced cancerogenesis, 10 min after the cells have been placed in a hypotonic medium, their size is equal to that at the point of maximal swelling, while the membrane potential is 20% lower than that in an isotonic medium. The reason for the varying adaptability of T-lymphocytes is, in our opinion, that, as the inflammation develops, the concentration of corticosteroids increases, the concentration of intracellular K+ decreases, the concentration of macromolecules increases and the potential of the plasmatic membrane of the cell rises, thus leading to the slowing down of the process of adaptation and its decreased extent. The lack of regulation of the volume by T-lymphocytes of mammals in the organisms of which cancerogenesis develops, testifies to the fact that, in such a case, cells lose their adaptability to changing external conditions. Therefore, the disappearance of adaptability of T-lymphocytes on the third day of development of cancerogenesis represents an early test of the process.


[Infrasound and biorhythms of the human brain]. A relationship is established between the frequency of the human brain alpha-rhythms and the frequency of infrasonic radiation emanating from overturned waves crests on the surface of seas and oceans. On this ground a recommendation is presented for maintaining the state of man's keeping awake.


[Theory of sound attenuation in the blood and erythrocyte suspensions]. Mechanisms of the ultrasound attenuation in blood and erythrocyte suspension in the long wave range are examined. It is shown that the theory proposed for dilute suspension of structured microobjects has a good coincidence with the known experimental data, both on erythrocyte suspensions and on blood. The contribution of viscous losses to attenuation are decreased with frequency and reach 44% in water suspensions of erythrocytes and 24% in the whole blood at 1 MHz.


[Thermal properties of chromatin in vivo during interaction with cis[Pt(CuO)2(CuOH)2]]. Thermal properties of chromatin in cell composition of BALB/c mice and whole chromatin at injection of cis-Pt(CuO)2(CuOH)2 to experimental animals were studied by high sensitive DSM assigning for investigating complex biological systems. It is established that injection of cis-Pt(CuO)2(CuOH)2 to leukemic mice in vivo changes the thermal characteristics of spleen cells, so that they look like the thermal properties of the healthy mice spleen cells. It was supposed that small doses of Pt(CuO)2(CuOH)2 in the case of leukemia show anticarcinogenic properties and the overdoses promote malignization; in the norm at any concentration of the injected salt cancerogenic properties are exhibited.


[Conformation of a triple collagen spiral as a function of primary structure]. It has been shown by the method of conformational analysis that conformation of the polypeptide chain of a collagen molecule varies from one type of sequence to another, but in all the sequences the only two types of conformations can be characterized by minimal conformational energy. The collagen molecule as a whole is a combination of the single-bond Rich and Crick model and new double-bonded structure. Rigorous comparison with "thick" conformations obtained by Scheraga and coworkers was undertaken using precise calculations with flexible proline.


[An ellipsometric study of the antigen-antibody interaction in the interphase solid/solution area]. Ellipsometric studies have proved that monoclonal immunoglobulin G(IgG) against gamma-interferon (gamma-INF) and immunoglobulin fraction (Ig-fraction) of rabbit blood serum against human serum albumin (HSA) are adsorbed according to the Langmuir model on the surfaces of mirror plates of covalently modified gamma-INF or HSA, respectively. The maximum surface concentrations (Tmax) and equilibrium adsorption constants (K) for IgG and Ig-fraction are equal to 2.57 pmol/cm2 and 2 x 10(7) M-1, 3.3 mg/m2 and 0.1 cm3/micrograms, respectively. The additional treatment of gamma-INF modified surfaces with Tween-20 leads to an increase of K IgG ut to 2.7 x 10(-7) M-1 while Tmax decreases up to 1.12 pmol/cm2 which is conditioned by the blocking of protein non-specific binding sites. The role of specific and non-specific interactions of IgG and Ig-fraction with covalently immobilized antigens was studied at antibody-antigen mixture adsorption. The necessity to apply this method to quantitative determination of gamma-IHF and HSA in solutions was proved.