Transesophageal echocardiographic assessment of left ventricular function in response to labetalol for control of postoperative hypertension. Although labetalol (LAB), the combination of an alpha- and beta-adrenergic blocking agent, is thought to be effective and safe for the control of postoperative hypertension, no study has focused on changes in left ventricular (LV) function when this drug was used to control postoperative hypertension. Therefore, this study determined the effects of LAB on hemodynamics and LV function assessed by 2D transesophageal echocardiography (TEE) in 17 patients undergoing abdominal aortic surgery who experienced a postoperative hypertensive episode. Postoperatively, patients were transferred while still intubated and under fentanyl sedation to the postanesthesia care unit where a TEE probe was inserted to provide a short-axis view of the LV. When their systolic blood pressure increased above 165 mmHg for more than 4 minutes, LAB was given in a dose of 0.75 mg/kg IV, over 2 minutes. If the blood pressure was not lowered to within 10% of the preoperative values, additional doses of LAB were given. Control of hypertension was obtained in all patients and was associated with a significant decrease in heart rate (90 +/- 19 to 70 + 13 bpm), cardiac index (4.52 +/- 1.65 to 3.36 +/- 1.55 L/min/m2), and mixed venous oxygen saturation (73 +/- 10 to 63 +/- 10%). With the lower blood pressure, end-diastolic area increased, indicating myocardial depression. In conclusion, LAB can be used to effectively control hypertension during the early postoperative period after abdominal aortic surgery. However, the reduction of blood pressure is achieved principally by the negative inotropic effect of LAB, which predominates over its vasodilator action.


Transesophageal Echo-doppler evaluation of the hemodynamic effects of positive-pressure ventilation after coronary artery surgery. Transesophageal echocardiography was used to extend knowledge about the impact of positive end-expiratory pressure (PEEP) during mechanical ventilation on right and left ventricular function and right ventricular impedance. At 20 cmH2O PEEP, a progressive increase of right ventricular end-diastolic area was seen (27%) that coincided with a reduction of early left ventricular filling velocity (25%) across the mitral valve, and a decrease of both pulmonary artery flow velocity (end-expiration 27% and end-inspiration 42%) and time-velocity index (end-inspiration 25%). As these changes were not accompanied by a change of the fractional area of contraction, the increase of the right ventricular diameter might be explained by right ventricular compensation due to an imbalance between augmented right ventricular impedance and reduced venous return.


Effects of fentanyl-diazepam-pancuronium anesthesia on heart rate variability: a spectral analysis. The effects of induction of anesthesia with fentanyl, 62.1 +/- 7.9 micrograms/kg, diazepam, 0.21 +/- 0.08 mg/kg, and pancuronium on beat-to-beat heart rate variations were studied in seven patients undergoing cardiac surgery. Spectral analysis was used to determine the intensity of the variations in each of the two main frequency bands in which variations are known to occur: (1) high-frequency band, mediated by the parasympathetic nervous system, at the frequency band of respiration, and (2) the low-frequency band, mediated by both sympathetic and parasympathetic nervous systems, at 0.04-0.12 Hz. Both the mean low- and high-frequency band areas were significantly decreased after induction of anesthesia. The ratio of high-to-low frequency band areas increased significantly after induction of anesthesia, from 45% to 62% (P less than 0.05). These results suggest that fentanyl-diazepam-pancuronium anesthesia decreases total autonomic nervous system activity and alters the balance between parasympathetic and sympathetic activities. Thus, spectral analysis of heart rate variations during anesthesia may provide a quantitative measure of assessing the activity of the autonomic nervous system.


The modified Fontan procedure: physiology and anesthetic implications. The modified Fontan operation has gained wide acceptance as a functional corrective procedure for patients with CHD with single ventricle physiology. Long-term survival and palliation of symptoms are excellent with most patients able to lead normal lives. The absence of a pulmonary contractile ventricle means that the single ventricle is responsible for perfusion of both the pulmonary and systemic circulations. Elevated systemic venous pressure is required to overcome PVR and this state of systemic venous hypertension has a significant impact on the anesthetic and postoperative care of these patients.


Pro: early endotracheal extubation is preferable to late extubation in patients following coronary artery surgery. Prolonged mechanical ventilation following CABG should not be uncritically considered "routine," and should only be used where indicated. A thorough physiologic and clinical evaluation with attention to hemodynamics, neurologic status, temperature and metabolism, hemostasis, and respiratory reserve should precede extubation. Continued post-operative ventilation is indicated in patients at high risk for complications, and it is possible to identify this subset preoperatively and upon arrival in the postoperative ICU. Early extubation (within 8 hours of arrival) should otherwise be the goal. The benefits of early extubation include improved cardiac function and patient comfort, reduction in respiratory complications, ease in management, and cost savings as the result of shortened length-of-stay in expensive postoperative units. More research is needed to clarify unanswered questions regarding ablating the stress response and avoiding myocardial ischemia.


Acute hemolysis in glutathione peroxidase deficiency. Severe acute hemoglobinemia and hemoglobinuria developed in a 17-year-old Japanese male with no history of exposure to any oxidizing agents. Examination of erythrocyte enzyme activity revealed that the patient's erythrocyte glutathione peroxidase (GSH-Px) activity was decreased to about a half that in control cells. Serum selenium (Se) concentration was within normal limits. Family studies showed that GSH-Px activity in the erythrocytes of two siblings was similarly decreased. It was suggested that the proband was suffering from a hereditary heterozygous GSH-Px deficiency, which appeared to be associated with acute hemolysis.


High serum human macrophage colony-stimulating factor level during pregnancy. Using a specific enzyme immunoassay, we monitored the serum levels of human macrophage colony-stimulating factor (hM-CSF) in pregnant women during gestation and after delivery. During pregnancy there was a marked elevation of maternal serum hM-CSF level, which returned to the baseline level within 3 weeks after delivery. The changes in maternal serum hM-CSF level were associated with changes in the numbers of monocytes and neutrophils in the maternal peripheral blood. Fetal sera prepared from cord blood samples also had a high hM-CSF level. In contrast to the baseline level of hM-CSF in maternal sera after delivery, the hM-CSF levels of newborn infants at 1 to 7 days after birth were higher than fetal levels, indicating that fetuses and newborn infants are a source of hM-CSF. The serum hM-CSF levels of infants at 22 to 35 days were between fetal levels and normal adult levels. Human M-CSF found in the serum of pregnant women and in cord blood was predominantly the large form of M-CSF, with a molecular mass of 85 kDa. Human M-CSF levels in amniotic fluid at 30 to 40 weeks were higher than those in cord blood sera at the same stage, indicating that the uterus is another source of hM-CSF. The roles played by hM-CSF during pregnancy need to be investigated.


Bone marrow transplantation for chronic myelogenous leukemia in blastic phase using a phenotypically identical unrelated volunteer donor. Nagoya Bone Marrow Transplantation Group (NBMTG), Tokai Marrow Donor Bank (TMDB). A 26-year-old male with chronic myelogenous leukemia in lymphoid blast crisis received a bone marrow transplant (BMT) from a phenotypically identical, mixed lymphocyte reaction (MLR)-weakly positive unrelated male volunteer donor. The volunteer was obtained from the Tokai Marrow Donor Bank (TMDB), which was established in Japan in 1989. This donor was selected from volunteer donors who were identical with our patient at the HLA-A,B loci, followed by matching at HLA-DQ, DR loci. On MLR testing, the donor's cells showed no response, but the patient's cells showed a low response to the donor's cells (relative response index 0.29). The patient showed rapid hemopoietic engraftment. He developed acute graft-versus-host disease (GVHD) with vesicle formation on palms and soles and mild liver damage, which were successfully treated with intravenous prednisolone 1 mg/kg per day. Although he also suffered from interstitial pneumonitis on day 64 and localized varicella-zoster infection on day 87, and has suffered from moderate stomatitis and dry skin characteristic of chronic GVHD, he is currently 22 months post-transplant with hematological remission and has a normal daily social life.


GM-CSF-mediated proliferation induction improves the susceptibility of leukemia cells to lymphokine-activated killer cells. Our studies on the susceptibility of fresh noncultured leukemia cells to interleukin-2 (IL-2)-induced lymphokine-activated killer (LAK) cells have shown that about two thirds of the leukemias are susceptible to the LAK-cell-mediated cytolysis. Analysis of 214 acute leukemias revealed considerable variation in the degree of cytotoxicity achieved. Cytolysis was substantially lower in fresh noncultured acute leukemia samples than in K562 and Daudi cell lines (mean Cr-release 21.0 +/- 16.0% versus 69.2 +/- 6.6% and 70.8 +/- 7.9%). Augmentation of susceptibility to LAK-cell lysis is desirable in connection with therapeutic application of IL-2-induced effector mechanisms. We observed a relationship between the LAK-cell susceptibility of leukemia cells and their spontaneous proliferation rate, which is significantly higher in LAK-cell-sensitive than in LAK-cell-resistant leukemias. It was therefore considered useful to examine the possibility of augmenting LAK-cell sensitivity by proliferation induction. These studies demonstrate that incubation of blast cells from acute myeloid leukemia (AML) and chronic myelogenous leukemia in myeloid blast crisis (CML-BC) with recombinant granulocyte-macrophage colony stimulating factor (GM-CSF) significantly augments LAK-cell susceptibility and that this is associated with an increased cell proliferation.


Colloidal gold uptake as a marker for monocyte differentiation and maturation in normal and leukemic cells. The uptake of colloidal gold particles by human monocytes was studied by electron microscopy, with special emphasis on changes in this uptake during the differentiation and maturation of these cells. The way in which leukemic cells of childhood acute non-lymphocytic leukemia (ANLL) can function in this gold uptake was also examined. In monocytes, microendocytosis was temperature-dependent; colloidal gold uptake increased as temperatures rose from 4 degrees C to 37 degrees C. It appeared that gold particles first adhered to the cell surface membrane, were then incorporated into the cytoplasmic vesicles, and then were transported into the granules. Original HL-60 cells and retinoic acid (RA)-treated HL-60 cells, which were differentiating and maturing along the granulocyte lineage, did not ingest colloidal gold particles, but 1,25(OH)2D3-treated HL-60 cells showed colloidal gold uptake during their differentiation and maturation along the monocyte lineage: 68.6% of the cells contained gold particles. Gold uptake was demonstrated in 27.3% of original U937 cells; the percentage increased to 70.3% when they were induced to mature by RA. In 15 specimens of childhood ANLL, none of the M1, M2 or M3 cells showed colloidal gold uptake, whereas 76-97% of M4 and M5 cells showed this uptake. These findings indicate that colloidal gold uptake is a marker of monocyte differentiation and maturation and can provide additional information for ANLL cytology.


Perivascular Ki-1 + lesions.

Perivascular Ki-1 + lesions. We describe three male patients diagnosed by histologic and immunophenotypic criteria to have a Ki-1 + lymphoproliferative disorder. All three cases shared a unique morphologic finding, not previously described: prominent perivascular cuffing of anaplastic/pleomorphic tumor cells around small and medium sized vessels. One case was a Ki-1 + anaplastic large cell lymphoma (ALCL) which developed in the setting of a low grade follicular B-cell lymphoma. A pseudoglandular pattern caused referral for consultation as a possible adenocarcinoma. One case was a cutaneous Ki-1 + lymphoproliferative disorder consistent with lymphomatoid papulosis (LyP). The third case was a cutaneous Ki-1 + ALCL. This study provides evidence that although perivascular cuffing of tumor cells is not frequently seen in Ki-1 + lymphoproliferative disorders (3 of 116 cases in our consultation file), it may be a prominent architectural feature causing confusion with epithelial or mesenchymal tumors. For this reason we recommend inclusion of this feature in the list of architectural features already described for Ki-1 + lymphoproliferative disorders.


A multicenter prospective study on the treatment of chronic idiopathic thrombocytopenic purpura. A multicenter prospective study on the treatment of chronic idiopathic thrombocytopenic purpura (ITP) was conducted by the Idiopathic Disorders of Hematopoietic Organs Research Committee (IDHORC), the Ministry of Health and Welfare of Japan. The aim of the study was to establish an improved therapeutic guide for chronic ITP. Of the 247 eligible patients 175 have been followed up to the present time, 16 patients have died, and 56 have been lost to follow-up. The median follow-up time was 55 months (range: 1 to 91 months). Of the 206 patients treated with corticosteroids, 13.1% achieved sustained complete remission. Splenectomy produced remissions in 52.5% of 72 patients evaluated 12 months after operation. Eighty-five patients were treated with immunosuppressive agents, but the response rates were low and the effect was transient in most cases. In the patients followed up without any specific treatment, the mean platelet counts showed slight improvement. Of the 16 patients who died during the observation period, only one died of bleeding; the remaining 15 died of causes unrelated to thrombocytopenia. It is noteworthy that five patients who were treated with corticosteroids or immunosuppressive agents died of infection.


Danazol therapy in idiopathic thrombocytopenic purpura: the efficacy of low-medium dose therapy. To examine the effects and optimal dose of danazol on idiopathic thrombocytopenic purpura (ITP), we administered a low-medium dose to 14 patients with this disease. A low-medium dose of danazol was effective in maintaining the platelet count at a high level, even after the dose of prednisolone was reduced. A low-medium dose of danazol without other therapy was effective in 3 of 6 patients even after they had been refractory to other treatment. A low dose of danazol was also effective in some patients for whom the other regimes were not indicated. It is concluded that a low-medium dose of danazol instead of a high dose is worth trying in ITP when the patient has become refractory to other therapeutic approaches or when these are not indicated.


Frequent clonal proliferation of human T-cell leukemia virus type 1 (HTLV-1)-infected T cells in HTLV-1-associated myelopathy (HAM-TSP). Human T-cell leukemia virus type 1 (HTLV-1) integrates its proviruses into random sites in host chromosomal DNA. Random integration of the proviruses was observed in asymptomatic HTLV-1 carriers and patients with HTLV-1-associated myelopathy (HAM/TSP). However, clonal integration has been reported in patients with adult T-cell leukemia (ATL), including that in the smoldering, chronic, and acute states, indicating clonal expansion of infected cells. In this study, we found that about 20% of HAM/TSP patients and their seropositive family members harbored subpopulation(s) of clonally proliferated cells infected with HTLV-1, although they still maintained randomly infected cells as a major population. These clones were stable during examination periods of 4 months to 3 years. However, these carriers or HAM/TSP patients did not show any significant indication of ATL. This extremely high frequency of clonal expansion of HTLV-1-infected cells indicates that some clones of HTLV-1-infected cells have a tendency to proliferate more efficiently than the other population without malignant transformation.


Simultaneous genotypic and immunophenotypic analysis of interphase cells using dual-color fluorescence: a demonstration of lineage involvement in polycythemia vera. Fluorescent in situ hybridization has become a useful technique by which chromosomal abnormalities may be shown in interphase cells. We present a dual-fluorescence method whereby a chromosomal and immunophenotypic marker can be visualized simultaneously in the same interphase cell. Two patients with the myeloproliferative disorder polycythemia vera and trisomy for chromosome 8 have been studied using this technique and selective involvement of the myeloid and erythrocyte lineages has been shown by the detection of the trisomy in immunophenotyped cells. Simultaneous analysis of genotype and immunophenotype in individual cells from patients with myeloproliferative disorders or leukemia may help identify the developmental and lineage status of cells in which molecular alterations have resulted in clonal advantage.


Blood platelets stimulate the expression of chondroitin sulfate proteoglycan in human monocytes. Mononuclear phagocytes synthesize chondroitin sulfate proteoglycan (CSPG), which is constitutively secreted. Because mononuclear phagocytes are known to interact with blood platelets, the effect of platelets on the release of CSPG in cultured human monocytes was investigated. After 6 days in vitro, the monocytes were supplied with fresh medium with different additions and subsequently exposed to [35S]sulfate for 24 hours before the medium fractions were harvested and analyzed for content of [35S]CSPG. Indirect evidence for the release of stimulatory factors from blood platelets was found when the addition of medium containing 50% serum made from platelet-rich plasma increased the expression of [35S]CSPG almost sevenfold compared with serum-free medium, whereas medium containing 50% serum made from platelet-depleted plasma increased the expression of [35S]CSPG about fourfold. Further, direct evidence for the stimulatory effect of platelets was found as the addition of autologous platelets to serum-free medium increased the expression of [35S]CSPG about threefold, and addition of supernatant from a corresponding number of thrombin-stimulated platelets was almost as efficient. The effect of five different platelet-derived factors (which are all present in serum) was investigated. Both platelet-derived growth factor (PDGF), platelet factor 4 (PF 4), and prostaglandin E2 (PGE2) used in physiologic concentrations were found to stimulate the expression of [35S]CSPG twofold to threefold, whereas transforming growth factor-beta had a slight inhibitory effect. 12-Hydroxyeicosatetraenoic acid had no significant effect on the expression of [35S]CSPG. Further evidence for the stimulatory effect of PDGF, PF 4, and PGE2 was found as serum depleted of these factors had significantly less stimulatory effect than control serum. The increased incorporation of [35S]sulfate into [35S]CSPG in cultures stimulated with serum or platelet-derived factors was not due to differences in molecular size or extent of sulfation of the proteoglycan molecules.


Spectrin beta Tandil, a novel shortened beta-chain variant associated with hereditary elliptocytosis is due to a deletional frameshift mutation in the beta-spectrin gene. An Argentinian family with hereditary elliptocytosis (HE) associated with a shortened beta-spectrin (Sp) chain was studied. As with most of the other shortened Sp beta-chains that have been described, this variant, called SpTandil, has impaired ability to participate in Sp dimer self-association, has lost its ability to become phosphorylated, and is associated with the presence of increased amounts of the alpha I 74-Kd fragment after partial tryptic digestion of Sp. The 3' ends of the beta-Sp gene of affected patients were analyzed. cDNA was prepared by reverse transcription of peripheral blood mRNA and amplified by the polymerase chain reaction (PCR) using primers corresponding to sequences 400 bp apart on the cDNA, spanning the last three exons (X, Y, Z) of the beta-Sp gene. Agarose gel electrophoresis of the cDNA amplification showed the presence of one band, the size of which was apparently the same as the band amplified from mRNA of a normal control. cDNA from one HE patient was subcloned and sequenced. Several clones showed the presence of a 7-bp deletion at codon 2041 in exon X. Genomic DNA of all the affected members of the family were amplified by PCR using primers flanking the deletion and corresponding to sequences 128 bp apart on exon X. Analysis of the PCR products using electrophoresis on polyacrylamide gel showed the presence of 121- and 128-bp bands in all HE subjects, and an additional doublet migrating more slowly than the two bands, which corresponded to the presence of heteroduplexes. The mutation results in a shift of the normal reading frame and leads to a new amino acid sequence at the C-terminus of the mutant beta-Sp chain. A new in-frame stop codon is encountered downstream, leading to premature chain termination. The identification of the molecular defect in Sp beta Tandil provides information regarding the region of the beta-Sp chain that is important for both Sp dimer self-association and an indication of potential sites of phosphorylation of the chain.


Bone marrows of non-Hodgkin's lymphoma patients with a bcl-2 translocation can be purged of polymerase chain reaction-detectable lymphoma cells using monoclonal antibodies and immunomagnetic bead depletion. Using the extremely sensitive technique of polymerase chain reaction (PCR) to detect the bcl-2 translocation, only 50% of bone marrows could be purged of PCR-detectable lymphoma cells using a cocktail of three anti-B-cell monoclonal antibodies (MoAbs) and complement-mediated lysis. This observation is of clinical importance because those patients whose reinfused marrows harbored residual lymphoma cells showed a significantly increased incidence of relapse. To improve purging, we used PCR detection of the bcl-2 translocation to compare the efficiency of complement-mediated lysis with immunomagnetic bead depletion. Using either a three or a four MoAb cocktail followed by immunomagnetic bead depletion, all PCR-detectable cells were purged after three cycles of treatment. In these same patient samples, treatment with three MoAbs and complement purged only 11 of the 25 (44%) samples. The addition of a fourth MoAb followed by complement lysis purged the marrows of only an additional five patients. Immunomagnetic bead depletion was specific because there was no loss of committed myeloid progenitor cells. The above results suggest that immunomagnetic bead depletion of the harvested marrow will likely be superior to our previous method of purging and the lack of nonspecific toxicity to myeloid progenitor cells predicts that it will not impair engraftment. This methodology will now be used to determine whether the reinfusion of lymphoma free marrow affects the incidence of relapse after autologous bone marrow transplantation.


Bone marrow transplants may cure patients with acute leukemia never achieving remission with chemotherapy. About 30% of adults with acute lymphoblastic leukemia (ALL) and 20% to 40% of children and adults with acute myelogenous leukemia (AML) never achieve remission, even with intensive chemotherapy. Most die of resistant leukemia, often within 6 months or less. In this study of 126 patients with resistant ALL or AML, allogeneic bone marrow transplants from HLA-identical siblings produced remissions in 113 of 115 (98%) evaluable patients. The 3-year probability of leukemia-free survival was 21% (95% confidence interval, 15% to 29%). Leukemia-free survival was similar in ALL (23%, 12% to 40%) and AML (21%, 14% to 31%). Only 3 of 27 patients at risk relapsed more than 2 years posttransplant.


Primary dexamethasone treatment of multiple myeloma. Intermittent courses of dexamethasone (DEX) were administered to 112 consecutive, previously untreated patients with multiple myeloma (MM). Using criteria based on a 75% or greater reduction of calculated tumor mass, the overall response rate was 43%. Among comparable patients, response rate were approximately 15% less than those observed previously with vincristine-doxorubicin by continuous infusion with intermittent DEX (VAD) and similar to those with melphalan-prednisone. The projected survival times with VAD or DEX were similar. Results indicated that DEX accounted for most of the plasma cell reduction achieved with VAD. Serious complications occurred in 27% of patients treated with VAD, but in only 4% of those who received DEX. In view of the similar outcome with fewer serious complications, DEX provided a simple, effective, and safe primary treatment for a large fraction of patients with MM. Patients who appear most likely to benefit include those with hypercalcemia or pancytopenia, or who require simultaneous radiotherapy for a pathologic fracture.


Polycythemia vera. II. Hypersensitivity of bone marrow erythroid, granulocyte-macrophage, and megakaryocyte progenitor cells to interleukin-3 and granulocyte-macrophage colony-stimulating factor. Polycythemia vera (PV) is a clonal disease of the hematopoietic stem cell characterized by a hyperplasia of marrow erythropoiesis, granulocytopoiesis, and megakaryocytopoiesis. We previously reported that highly purified PV blood burst-forming units-erythroid (BFU-E) are hypersensitive to recombinant human interleukin-3 (rIL-3). Because these cells may be only a subset, and not representative of marrow progenitors, we have now studied partially purified marrow hematopoietic progenitor cells. Dose-response experiments with PV marrow BFU-E showed a 38-fold increase in sensitivity to rIL-3 and a 4.3-fold increase in sensitivity to recombinant human erythropoietin (rEpo) compared with normal marrow BFU-E. In addition, PV marrow colony-forming units-granulocyte-macrophage (CFU-GM) and CFU-megakaryocyte (CFU-MK) also showed a marked hypersensitivity to rIL-3 and to human recombinant granulocyte-macrophage colony-stimulating factor (rGM-CSF). Dose-response curves with rGM-CSF and blood BFU-E showed a 48-fold increase in sensitivity. No effect of rIL-4, rIL-6, human recombinant granulocyte-CSF (rG-CSF), or macrophage-CSF (rM-CSF) was evident, nor was there any effect of PV cell-conditioned medium on normal BFU-E, when compared with normal cell-conditioned medium. Autoradiography with 125I-rEpo showed an increase in Epo receptors after maturation of PV BFU-E to CFU-E similar to that shown with normal BFU-E, but no increase of specific binding of 125I-rIL-3 by PV CD34+ cells was seen compared with normal CD34+ cells. These studies show that PV marrow hematopoietic progenitor cells are hypersensitive to rIL-3 and rGM-CSF, similar to PV blood BFU-E. While the mechanism does not appear to be due to enhanced binding of rIL-3, the hypersensitivity of PV progenitor cells to IL-3 and GM-CSF may be a key factor in the pathogenesis of PV.


Binding of primitive hematopoietic progenitor cells to marrow stromal cells involves heparan sulfate. Blast colony-forming cells (BI-CFC) and pre-colony-forming unit-granulocyte, monocyte (CFU-GM) in human bone marrow bind to marrow-derived stromal layers grown in the presence of methylprednisolone (MP+), but do not bind to stroma grown without MP (MP-). The BI-CFC bind to stroma and form colonies when overlaid with agar; the pre-CFU-GM bind to stroma and release CFU-GM into the supernatant culture medium (delta assay). These two classes of progenitor may represent similar stages of hematopoietic cell development. Their binding to stroma depends on the presence of heparan sulfate proteoglycan (HS-PG) in the extracellular matrix secreted by the stromal cells. Here, we have analyzed the functional and biochemical properties of HS-PG isolated from MP+ and MP- stromal cultures. HS-PG or isolated HS glycosaminoglycan (GAG) side chains partially blocked progenitor cell binding when they were added to the 2-hour binding phase of the BI-CFC or delta assays. Gel electrophoresis of HS-PG resolved more bands in matrix preparations from MP+ cultures than in preparations from MP- cultures. The blocking activity of the eluted MP+ HS-PG bands depended partly on the amount of GAG attached to the protein core and presumably partly on the structure of the core itself. Time course studies demonstrated that the HS-dependent phase of the binding interaction was limited to the first 30 to 60 minutes of the 2-hour binding phase. The different blocking effects of MP+ and MP- HS indicate that they have different biochemical properties. The HS-GAG in MP+ stroma has a higher degree of sulfation and a greater negative charge to mass ratio compared with MP- HS-GAG. Variations in HS may determine specific binding by hematopoietic progenitor cells and a heparan sulfate receptor is envisaged as acting in concert with further cell adhesion molecules (CAMs) on the progenitor cell surface.


Role of individual gamma-carboxyglutamic acid residues of activated human protein C in defining its in vitro anticoagulant activity. To evaluate the contributions of individual gamma-carboxyglutamic acid (gla) residues to the overall Ca(2+)-dependent anticoagulant activity of activated human protein C (APC), we used recombinant (r) DNA technology to generate protein C (PC) variants in which each of the gla precursor glutamic acid (E) residues (positions 6, 7, 14, 16, 19, 20, 25, 26, and 29) was separately altered to aspartic acid (D). In one case, a gla26V mutation ([gla26V]r-PC) was constructed because a patient with this particular substitution in coagulation factor IX had been previously identified. Two additional r-PC mutants were generated, viz, an r-PC variant containing a substitution at arginine (R) 15 ([R15]r-PC), because this particular R residue is conserved in all gla-containing blood coagulation proteins, as well as a variant r-PC with substitution of an E at position 32 ([F31L, Q32E]r-PC), because gla residues are found in other proteins at this sequence location. This latter protein did undergo gamma-carboxylation at the newly inserted E32 position. For each of the 11 recombinant variants, a subpopulation of PC molecules that were gamma-carboxylated at all nonmutated gla-precursor E residues has been purified by anion exchange chromatography and, where necessary, affinity chromatography on an antihuman PC column. The r-PC muteins were converted to their respective r-APC forms and assayed for their amidolytic activities and Ca(2+)-dependent anticoagulant properties. While no significant differences were found between wild-type (wt) r-APC and r-APC mutants in the amidolytic assays, lack of a single gla residue at any of the following locations, viz, 7, 16, 20, or 26, led to virtual complete disappearance of the Ca(2+)-dependent anticoagulant activity of the relevant r-APC mutant, as compared with its wt counterpart. On the other hand, single eliminations of any of the gla residues located at positions 6, 14, or 19 of r-APC resulted in variant recombinant molecules with substantial anticoagulant activity (80% to 92%), relative to wtr-APC. Mutation of gla residues at positions 25 and 29 resulted in r-APC variants with significant but low (24% and 9% of wtr-APC, respectively) levels of anticoagulant activity. The variant, [R15L]r-APC, possessed only 19% of the anticoagulant activity of wrt-APC, while inclusion of gla at position 32 in the variant, [F31L, Q32gla]r-APC, resulted in a recombinant enzyme with an anticoagulant activity equivalent to that of wtr-APC.


Role of plasma viscosity in platelet adhesion. Platelet adhesion to the vessel wall is initiated by transport of blood platelets from the bulk flow to the wall. The process of diffusion and convection of the platelets is affected by rheological conditions such as well shear rate, red blood cell (RBC) deformability, and viscosity of the medium. To study the effect of plasma viscosity on platelet adhesion, perfusion experiments with a rectangular perfusion chamber were performed. Reconstituted blood, consisting of washed platelets and washed RBCs, was circulated through this chamber for 5 minutes at a wall shear rate of 300 s-1. Different albumin concentrations were made, to obtain different medium viscosities (0.89 to 1.85 mPa.s). Platelet adhesion decreased with increasing medium viscosity up to viscosities of 0.95 mPa.s, but increased with medium viscosity above this value. Instead of human albumin solution, different plasma viscosities were obtained by dilution of Waldenström plasma with buffer. Plasma was depleted of fibronectin, which gave a final plasma viscosity of 2.0 mPa.s, and was dialyzed against HEPES buffer and subsequently diluted with the dialysis buffer in different fractions (0.89 to 2.00 mPa.s). Perfusions were performed over a purified von Willebrand factor coating on glass, or over an endothelial cell matrix, preincubated with von Willebrand factor. With both surfaces, platelet adhesion was dependent on the plasma viscosity in a similar way: at low plasma viscosities, adhesion was decreased with increasing plasma viscosity, while at higher plasma viscosities, adhesion increased with plasma viscosity. Adhesion values at higher plasma viscosity or at higher human albumin concentrations could be explained by effects of the medium on the rigidity of the RBCs, since platelet adhesion is known to be increased by enhanced RBC rigidity. Effects of the medium on the deformability of the RBCs were measured separately with the laser diffraction method. These experiments confirmed that presence of human albumin or plasma in the measuring suspension increased the rigidity of RBCs. To prevent influence of the medium on the RBCs in perfusion experiments, the RBCs were fixated with glutaraldehyde. Perfusion experiments with fixated RBCs in plasma over a von Willebrand factor preincubated endothelial cell matrix, showed a consequent decrease in adhesion with increasing plasma viscosity, according to the diffusion theories, whereas the increase of adhesion at high plasma viscosities was lacking. This suggests that the latter effect was entirely due to increased transport of platelets by more rigid RBCs.


Sustained antithrombotic activity of hirudin after its plasma clearance: comparison with heparin. Thrombus extension in patients with venous thromboembolism is due to the accretion of fibrin onto existing thrombi. Extension is promoted by both circulating and thrombus-bound thrombin, which convert fibrinogen to fibrin. Heparin is an effective antithrombotic agent, but it requires continuous administration to achieve persistent inhibition of thrombus extension. Heparin is highly effective in inhibiting fluid phase thrombin, but is a relatively ineffective inhibitor of thrombus-bound thrombin. Hirudin, unlike heparin, inactivates both circulating and fibrin-bound thrombin and, therefore, has the potential to prevent thrombus extension even after a short course of treatment. The aim of this study was to evaluate the time course of the accretion of new fibrin onto preexisting rabbit jugular vein thrombi after a 3-hour infusion of saline, heparin, and hirudin. Heparin and recombinant (r)-hirudin (CGP 39399) were infused at doses that doubled the activated partial thromboplastin time (aPTT). At the end of the 3-hour infusions in rabbits treated with saline, heparin (0.75 mg/kg), or r-hirudin (1.25 mg/kg), accretion of 125I-fibrinogen was 59 +/- 5 micrograms, 34 +/- 4 micrograms, and 21 +/- 2 micrograms, respectively (heparin and r-hirudin v saline, P less than .01; r-hirudin v heparin, P less than .01). Three hours after the end of the infusions, the accreted 125I-fibrinogen in the saline-, heparin-, and hirudin-treated animals was 89 +/- 6 micrograms, 51 +/- 7 micrograms, and 23 +/- 3 micrograms, respectively; 9 hours after the end of the infusions, the accreted 125I-fibrinogen was 112 +/- 9 micrograms, 82 +/- 7 micrograms, and 25 +/- 3 micrograms, respectively. aPTT and thrombin clotting time (TCT) returned to the baseline value 90 minutes after the end of heparin or r-hirudin infusion. During in vitro experiments, human fibrin clots previously incubated in human plasma containing r-hirudin did not promote fibrinopeptide A (FPA) generation when washed and then incubated in human plasma in the absence of thrombin inhibitors. This persistent inhibition of FPA production was not observed after incubation in human plasma of human plasma clots preincubated with heparin. We conclude that heparin is effective in inhibiting thrombus extension while it is present in the circulation, but that this effect is rapidly lost after its plasma clearance. In contrast, the antithrombotic activity of r-hirudin is sustained beyond its plasma clearance, presumably because of its ability to inactivate thrombus-bound thrombin. Our findings indicate that r-hirudin might be an effective antithrombotic agent even when used for short periods.


Tumor necrosis factor-induced endothelial tissue factor is associated with subendothelial matrix vesicles but is not expressed on the apical surface. Cultured endothelial cells can be induced by tumor necrosis factor/cachectin (TNF) and other cytokines to synthesize the procoagulant cofactor tissue factor (TF). Intact monolayers of TNF-treated endothelial cells showed only minimal TF activity. In contrast, after permeabilization of these monolayers with detergent (saponin, 0.02%), there was approximately 10- to 20-fold increase in TF-mediated, factor VIIa-dependent factor Xa formation. Extracellular matrix derived from TNF-treated endothelium, prepared after removing the cells by hypotonic lysis or ammonium hydroxide (0.1 N), also had similarly enhanced TF activity. Incubation with a blocking monoclonal antibody to TF inhibited the procoagulant activity of both TNF-stimulated endothelial cells, whether they were intact or permeabilized, and of their matrices. However, when the apical cell surface was pretreated with anti-TF antibody, washed, and then cells were lysed with water or permeabilized with saponin, similar augmentation of TF activity was still observed, suggesting the presence of a pool of TF to which the antibody did not initially gain access. Consistent with this concept, the presence of TF in the matrix of TNF-treated endothelial cells was shown by immunoblotting and morphologic studies; cultured endothelial monolayers and the native endothelium of aortic segments after exposure to TNF showed TF in extracellular matrix, associated with vesicles. In contrast, TF was virtually undetectable on the apical endothelial surface. Taken together, these findings suggest that endothelial TF can be present in a cryptic pool that only gains access to the blood after alteration in the integrity of the endothelial monolayer.


Relationship of antiphospholipid antibodies to pregnancy loss in patients with systemic lupus erythematosus: a cross-sectional study. To determine whether an association exists between the presence of antiphospholipid antibodies and pregnancy loss, a cross-sectional study was performed. Consecutive women who were referred to three outpatient rheumatology clinics and who had systemic lupus erythematosus (SLE) and a history of one or more pregnancies were evaluated. Patients were interviewed to determine outcomes of all previous pregnancies. Blood was taken on two separate occasions at least 3 months apart to test for the presence of the lupus anticoagulant and anticardiolipin antibodies; on both occasions, five tests of the lupus anticoagulant, with well-defined normal ranges, and an enzyme-linked immunosorbent assay to measure IgG anticardiolipin antibodies were performed. Patients were considered to be positive for the lupus anticoagulant if one or more tests was abnormal on both occasions and positive for anticardiolipin antibodies if the test was abnormal on both occasions. Forty-two women were studied. Statistically significant associations were shown between lupus anticoagulant positivity and previous pregnancy loss (odds ratio [OR], 4.8; 95% confidence intervals [CI], 1.0 to 23.6; P = .05) and between anticardiolipin antibody positivity and previous pregnancy loss (OR, 20.0; 95% CI, 1.3 to 97.0; P = .01). All seven women with multiple episodes of pregnancy loss were lupus anticoagulant positive and four of these were also anticardiolipin antibody positive. If patients who are transiently positive for lupus anticoagulant and/or anticardiolipin antibodies are considered to be test positive, the associations with pregnancy loss are no longer statistically significant. Within the group of lupus anticoagulant-positive patients, we observed stronger associations between the presence of six or more positive tests and pregnancy loss than between the presence of two to five positive tests and pregnancy loss. No single test for the lupus anticoagulant provides a statistically significant association with pregnancy loss. The results of our study show that by performing multiple lupus anticoagulant tests and by repeating testing for lupus anticoagulant and anticardiolipin antibodies on more than one occasion, significant associations between the presence of antiphospholipid antibodies and previous pregnancy loss can be shown in patients with SLE.


Shear rate-dependent impairment of thrombus growth on collagen in nonanticoagulated blood from patients with von Willebrand disease and hemophilia A. Thrombus formation on collagen fibrils was quantified at venous (100/s) and arterial (650/s and 2,600/s) wall shear rates in blood from patients with various subtypes of von Willebrand disease (vWD) and with hemophilia A (HA). Nonanticoagulated blood was drawn directly from an antecubital vein over purified type III collagen fibrils exposed in parallel-plate perfusion chambers. Blood-collagen interactions were differentiated and quantified by morphometry as platelet adhesion, thrombus height, thrombus volume, and deposition of fibrin strands. Sixteen patients with vWD, including four type III, six type I, four type IIA, and two type IIB, were compared with 26 normal subjects and nine patients with HA, including six severe HA and three mild HA. Platelet adhesion and thrombus formation at 2,600/s were significantly decreased in blood from patients with vWD type III, IIA, and IIB, but not in blood from patients with type I and in HA. The abnormal thrombus formation was apparently not related to the decreased levels of factor VIII (F.VIII), because thrombus height and volume were normal in severe and mild HA. Thrombus formation at 650/s was also significantly decreased in patients with vWD type III, IIA, and IIB and slightly reduced in type I. Significant reduction in thrombus volume and height was also observed in blood from patients with severe HA, but not in mild HA. Thrombus dimensions were not affected at 100/s in the vWD subtypes. However, significantly decreased thrombus height and virtually absent fibrin deposition were observed in blood from patients with severe HA. Apparently, F.VIII supports thrombus formation at low and intermediate shear conditions, presumably through the generation of thrombin. In contrast, von Willebrand factor (vWF) mediates not only platelet adhesion, but also thrombus formation at intermediate and high shear rates. Thus, the relative contribution of coagulation (F.VIII) and platelet function (vWF) in thrombus formation appears to be shear rate dependent, but having optimal effects at different shear conditions.


A simple method for the quantitative evaluation of neuromuscular blockade in mice. The purpose of this study was to develop a simple and noninvasive method for the quantitative evaluation of neuromuscular blockade in mice. The sciatic nerve in anesthetized animals (etomidate, 50 mg/kg or mephenesin, 250 mg/kg) was stimulated using transcutaneous electrodes; repetitive supramaximal stimulations (3 Hz during 3.3 sec) were applied each minute until recovery or death occurred. Evoked action potentials of the gastrocnemius muscle were recorded. The effect of each pharmacological agent was expressed as the ratio (S5/S1) of the area of the fifth response (S5) to the first one (S1). The performance of the method was tested using neuromuscular blocking agents such as alcuronium, suxamethonium, or alpha-bungarotoxin. It was concluded that the method exhibits the required statistical performances (sensitivity, repeatability, and specificity) to be recommended for in vivo investigation of neuromuscular impairment.


Antipyrine as a dialyzable reference to correct differences in efficiency among and within sampling devices during in vivo microdialysis. Antipyrine was investigated as a dialyzable substance that could be used to quantitate relative differences in the efficiency of dialysis among multiple microdialysis probes and by a single probe over time. The contribution of effective membrane surface area to recovery variability was tested by the introduction of air into microdialysis probes. Reduction of effective membrane surface area reduced antipyrine recovery. Dialysates from probes implanted in the jugular vein, brain, and liver of rats receiving antipyrine demonstrated differences in antipyrine concentration among probes within the same rat. These results suggest dissimilar efficiencies of the probes to recover antipyrine, which should be uniformly distributed throughout body water. Dialysates from blood, brain, and liver probes in rats that received both antipyrine and tritiated water (3H2O) showed differences in antipyrine and 3H2O concentrations among probes. Variability of antipyrine and 3H2O concentrations over time within a probe were positively correlated, suggesting that the cause(s) of temporal variability affected both of these markers of body water. Correction of antipyrine tissue/blood ratios, using 3H2O blood/tissue ratios from the same sampling period, reduced the variability in antipyrine tissue/blood ratios, producing ratios closer to the expected value of 1. Differences in probe efficiency contributing to the variability of antipyrine and 3H2O recovery would also be expected to influence the recovery of other substances during microdialysis. The administration of antipyrine during microdialysis experiments is suggested to enable reduction of temporal and site-related differences in substance recovery that are due to differences in probe efficiency. Other methods are necessary to determine the actual extracellular concentration of dialyzed substances and the integrity of the blood-brain barrier.


Techniques for drug delivery to the airways, and the assessment of lung function in animal models. A common approach to understanding the mechanisms underlying clinical asthma and in new drug development is to mimic the disease in animal models. When developing animal models of pulmonary diseases, such as asthma, the experimentally induced disease may be characterized in terms of pathophysiological changes induced (e.g., inflammation, smooth muscle contraction) or by the indices of lung function that are effected by such changes. Similarly, the effects of drugs can be assessed in terms of the reversal of disease- or mediator-induced changes in lung function. Small animals, such as the guinea pig and rat, are commonly used for the assessment of lung function in models of pulmonary diseases, such as asthma, and to evaluate the effects of drugs. A variety of techniques, differing in their level of sophistication, has been developed to measure parameters of lung function in small laboratory animals. Simple techniques involve the visual assessment of the response of a conscious animal to bronchoconstriction induced by an inhaled spasmogen or antigen. This technique is rapid but gives results that are difficult to interpret in physiological terms. Bronchospasm can be better assessed in anesthetized, mechanically ventilated animals by recording bronchial tone as changes in either 1) ventilation circuit pressure or 2) air overflow as the lungs are inflated. These techniques are widely used but because they require surgical intervention they are not suited to long-term or repeat studies. In addition, they give only a limited indication of the physiological changes that affect airway caliber. To improve the models available, researchers have subsequently developed techniques that use the same physiological principles as some of the tests applied to the assessment of lung function in humans. These techniques allow the measurement of parameters of respiratory mechanics, such as lung compliance and airway resistance, that determine the relationship between pulmonary pressure changes and air flow into and out of the lungs. Continued development has resulted in models that use nonsurgical plethysmographic techniques. These allow the long-term or repeated measurement of lung function in conscious animals under minimal restraint. In the treatment of asthma, inhalation is the preferred route of administration of a drug as it allows rapid drug delivery to the site of action. Systemic effects are reduced, and the therapeutic dose is minimized. Drugs are generally inhaled as either nebulized liquids or dry-powder formulations. Because drug inhalation requires patient cooperation, techniques have been modified to allow drug delivery to the airways of experimental animals.(ABSTRACT TRUNCATED AT 400 WORDS)


The pathogenesis of DOCA-salt hypertension. Deoxycorticosterone acetate (DOCA) is an agent commonly used to induce hypertension in experimental animals. This form of hypertension is dependent on altered regulation of central pressor mechanisms including the brain renin-angiotensin system. Additionally, there are characteristic changes involving the cardiovascular system and baroreflex responses. This review will discuss aspects of the pathogenesis of DOCA hypertension and the effect of various antihypertensive agents on the development of this form of hypertension.


Measurement of cardiac left ventricular pressure in conscious rats using a fluid-filled catheter. A fluid-filled catheter consisting of 100 cm of PE50 polyethylene tubing welded to 7 cm of PE10 polyethylene tubing (PE50/PE10) was constructed for the purpose of measuring the rate of left ventricular pressure development (+dP/dt) in conscious, freely moving rats. Prior to in vivo experiments, four PE50/PE10 catheters were randomly selected, and their natural frequencies and damping ratios were determined using a square wave impact. The mean (n = 4), natural frequency of these catheters was shown to be 35.0 +/- 5.5 Hz, and the mean damping ratio was 0.83 +/- 0.10. Natural frequency plotted against increasing PE50 tubing length was shown to have a slope of -0.44 Hz/cm with a correlation coefficient of 0.99. The effect of the 7-cm PE10 tubing segment on the catheter damping ratio was also demonstrated. One of the four PE50/PE10 type catheters exhibited a damping ratio of 0.74 +/- 0.09. When the 7-cm PE10 tube was removed, the damping ratio was reduced to 0.31 +/- 0.04. Left ventricular +dP/dt obtained in conscious rats with a PE50/PE10 catheter (n = 7; 6300 +/- 300 mmHg/sec) was significantly less than the +dP/dt obtained using a 100-cm PE50 catheter (n = 6; 9400 +/- 400 mmHg/sec). The results of this study make it clear that the PE50/PE10 catheter is suitable for the measurement of left ventricular +dP/dt in the conscious rat, and that catheter design has a profound influence on both the catheter natural frequency and damping ratio.


Chronic intrathecal catheterization in the sheep. This paper describes a relatively simple and noninvasive method for the chronic implantation of intrathecal catheters in the sheep. The technique has been carried out on 17 occasions in nine sheep, with 60% of attempted catheterizations producing a correctly positioned, functional catheter. The placement and integrity of the catheters were confirmed by radiography using a contrast medium. Correctly placed catheters have been maintained for up to 16 months without problems.


Epicardial His bundle recordings in the guinea pig in vivo. In small animals, His bundle activity is commonly registered using intracavitary electrodes in Langendorff-perfused hearts. The present study evaluates the feasibility of epicardial registration of His bundle activity in 30 guinea pigs in vivo, anesthetized with 60 mg/kg sodium pentobarbital i.p. Following median sternotomy, a bipolar electrode, mounted on a flexible stand, was placed into the aortic-right atrial groove. His bundle activity was easily detected in all animals. Mean PA, AH, and HV intervals were 16.7 +/- 1.4 msec (range: 6.0-34.0 ms), 36.4 +/- 1.5 ms (range: 22.0-54.0 msec) and 14.4 +/- 0.4 msec (range: 9.0-20 msec), respectively. Administration of verapamil and vagal stimulation induced significant prolongations of AH intervals without affecting HV intervals; lidocaine prolonged both AH and HV intervals. Both at baseline and following drug administration there was excellent agreement between the AH and HV intervals assessed from simultaneously registered epicardial and endocardial His bundle electrograms (n = 12). The presented model provides reproducible values for atrioventricular (AV) nodal and His-to-ventricular conduction intervals in the guinea-pig in vivo and will, thus, be a valuable tool in electrophysiologic drug evaluation.


Technique of reflection electron microscopy. Details of the technique of reflection electron microscopy (REM) are described. Step by step instruction is given on how to do it on an ordinary electron microscope. Also given are some specimen preparation techniques and strategy of REM investigation.


Recent studies of surface dynamic processes by reflection electron microscopy. Reflection electron microscope studies of surface dynamic processes are reviewed and illustrated with recent new observations. They include: surface electromigration and current dependent structures of Si surfaces; surface etching by oxidation of Si surface; and growth of two dimensional alloyed adsorbate by co-deposition of metals on Si surface. The observations revealed details of the surface dynamic processes, which are difficult to obtain with other surface analysis techniques.


Application of ultrahigh vacuum reflection electron microscopy for the study of clean silicon surfaces in sublimation, epitaxy, and phase transitions. The construction and performance of an ultrahigh vacuum reflection electron microscope (UHV REM) on the base of a transmission electron microscope with top entry stage are described. Some results of in situ study of structural transformations on clean silicon surfaces during sublimation, surface phase transitions, and initial stages of epitaxial growth are shown.


Degradation, amorphization, and recrystallization of ion bombarded Si(111) surfaces studied by in situ reflection electron microscopy and reflection high energy electron diffraction techniques. In this paper we report the effect of noble gas ions bombardment on the degradation of atomically flat Si(111) surfaces at room and high (400 degrees C-600 degrees C) temperatures. Reflection high energy electron diffraction (RHEED) and reflection electron microscopy (REM) have been used to characterize the topography and structure of the as-implanted and post annealed surface layers. It is shown that the fading of the specularly reflected beam is not directly related to the amorphization of the surface. This experimental study has also evidenced the difficulties one meets to regrow a defect-free material after amorphization by noble gas bombardment. For high temperature for which the amorphization is not possible, the surface loses its stepped structure and turns into a monocrystalline but atomically rough surface. This roughness is a function of substrate temperature.


Bloch wave treatment of symmetry and multiple beam cases in reflection high energy electron diffraction and reflection electron microscopy. Bloch wave equations for the multiple beam cases in reflection high energy electron diffraction (RHEED) are derived from the integral equation by forward- and back-scattering Green function operators. A linearization is achieved through separation in a forward- and a back-scattering component for each beam. This leads to a set of fundamental equations similar to the transmission case, but with a non-Hermitian matrix, and the beams may be entered as either forward-, back-scattered, or both. The number of beams needed to be included in RHEED calculations is thus reduced, and so are the computing time and computer space required. The systematic row case, corresponding to reflections from planes parallel to the crystal surface, is treated in detail and illustrated by calculations of dispersion surfaces and rocking curves for Au(001). Symmetry relations for the systematic row and between reciprocal rows are discussed and illustrated.


Developments in the dynamical theory of high energy electron reflection. High energy electron reflection (HEER) is an important technique in surface science and uses the information carried by high energy electrons reflected from surfaces to study surface structures and surface electronic states. With the development of reflection high energy electron diffraction (RHEED), high energy electron microscopy (REM), and high energy electron energy loss spectroscopy (EEL) in surface science, the usefulness of HEER has been widely recognized and demonstrated. However, a stationary dynamical solution for an arbitrary surface for HEER has not been obtained yet. In this paper, some developments in understanding the dynamical theory of HEER, particularly in recent years, are reviewed: 1. The introduction of the concept of current flow for a semi-infinite crystal model has removed the confusion around the wave points in the "band gap." 2. The consistency between the Bloch wave and multislice in the Bragg case has verified the validity of the argument of current flow and led to the emergence of the BMCR method (Bloch wave + Multislice Combined for Reflection). 3. The failure of the Bloch Wave-Only solution (the BWO solution) on Au (110) surfaces in the Bragg case revealed by the BMCR method implies that previous BWO calculations in the Bragg case might be at fault. 4. The 2-D dependence of the electron wave fields and Picard iteration-like character of multislice calculation in the Bragg case has led to the emergence of an Edge Patching method in Multislice-mode-Only (the EPMO method). The new method yields an infinitely convergent stationary dynamical solution for an arbitrary surface.


Reflection electron energy-loss spectroscopy and imaging for surface studies in transmission electron microscopes. A review is given on the techniques and applications of high-energy reflection electron energy-loss spectroscopy (REELS) and reflection electron microscopy (REM) for surface studies in scanning transmission electron microscopes (STEM) and conventional transmission electron microscopes (TEM). A diffraction method is introduced to identify a surface orientation in the geometry of REM. The surface dielectric response theory is presented and applied for studying alpha-alumina surfaces. Domains of the alpha-alumina (012) surface initially terminated with oxygen can be reduced by an intense electron beam to produce Al metal; the resistance to beam damage of surface domains initially terminated with Al+3 ions is attributed to the screening effect of adsorbed oxygen. Surface energy-loss near-edge structure (ELNES), extended energy-loss fine structure (EXELFS), and microanalysis using REELS are illustrated based on the studies of TiO2 and MgO. Effects of surface resonances (or channeling) on the REELS signal-to-background ratio are described. The REELS detection of a monolayer of oxygen adsorption on diamond (111) surfaces is reported. It is shown that phase contrast REM image content can be significantly increased with the use of a field emission gun (FEG). Phase contrast effects close to the core of a screw dislocation are discussed and the associated Fresnel fringes around a surface step are observed. Finally, an in situ REM experiment is described for studying atomic desorption and diffusion processes on alpha-alumina surfaces at temperatures of 1,300-1,400 degrees C.


Observation of atomic steps on single crystal surfaces by a commercial scanning electron microscope. Atomic steps on (111) and (100) crystal surfaces of Pt were observed using a commercial scanning electron microscope (SEM) in secondary electron mode. By comparing the SEM images and those by reflection electron microscopy (REM), the observed contrast was confirmed to be that from atomic steps on crystal surfaces. The contrast mechanism is briefly discussed. One application of this imaging technique is also shown.


Observation of double line contrast in surface imaging. The double line contrast of a single-atom height step observed in surface imaging for a single crystal in reflection electron microscopy is studied under a variety of experimental conditions. It is suggested that this abnormal contrast is directly associated with the dynamical electron diffraction process. The behavior of the double line contrast is closely related to the order of the Bragg reflected beam, and can be observed mostly under one of the two commonly cited resonance conditions. This phenomenon clearly reveals the differences in the surface imaging for various resonance conditions.


Preparation and characterization of MgO surfaces by reflection electron microscopy. We have employed several different methods to prepare (100) and (111) surfaces of MgO crystals. (100) surfaces prepared by simple cleaving give good reflection high energy electron diffraction (RHEED) patterns and surfaces with a high density of coarse steps. Chemical polishing of this surface results in a roughening of the topography whilst annealing in oxygen considerably smoothens the surfaces although they appear to be contaminated. Under certain conditions we find that the MgO crystals will cleave along the (111) plane. Both cleaved and mechanically polished (111) surfaces are atomically flat and reconstructed after oxygen annealing.


Characterization of the annealed (0001) surface of sapphire (alpha-Al2O3) and interaction with silver by reflection electron microscopy and scanning reflection electron microscopy. Annealed (0001) surfaces of single-crystal sapphire (alpha-Al2O3) rod have been studied in the electron microscope using reflection electron microscopy (REM), scanning reflection electron microscopy (SREM), and reflection high energy electron diffraction (RHEED). Annealed surfaces of (0001) sapphire are vicinal and characterized by close-packed (0001)-oriented terraces separated by faceted multiple-height steps, with edges parallel to energetically preferred low-index directions (less than 1010 greater than and less than 1120 greater than). These structural features are not seen on cleaved surfaces or polished surfaces treated at temperatures less than 1,250 degrees C. Oxygen-annealing produces clean surfaces which prove useful for investigating the interaction of deposited metals with the (0001) sapphire. Both REM and SREM (with microdiffraction spots) techniques have been used to observe fine structure of flat Ag islands on the scale of 1-100 nm on the (0001)-oriented terraces as well as aggregations at the steps. A preliminary result on interaction with Cu is also included.


Reflection electron microscopy and interferometry of atomic steps on gold and platinum single crystal surfaces. Two beam interferences produced using an electrostatic biprism, which is inserted in the position of the selected area diaphragm of a commercial electron microscope, may be used in reflection electron microscopy to determine the phase shifts induced by structures on single crystal surfaces. A description of our interferometrical and holographical experiments on the phase shift at steps on (111)Au and (111)Pt single crystal surfaces is given and a straight forward interpretation of the results in terms of refraction will be discussed. As a particular result phase shifts of pi and 0.9 pi were measured for monatomic steps on (111)gold and (111)platinum surfaces, respectively.


Observation of surfaces by reflection electron holography. Reflection electron holography is described as a method to observe sub-A surface morphology. Phase shift of a Bragg-reflected electron wave was measured by means of holographic interferometry using an electron microscope equipped with a field emission electron gun and an electron biprism. A short wavelength of high energy electrons is the essential key to the high vertical sensitivity of this method, since geometrical path differences produced by the surface topography are measured in units of wavelengths in interferometrical measuring. Phase shift at a monoatomic step and the displacement field around a dislocation emerging on the surface were observed.


SpCoel1: a sea urchin profilin gene expressed specifically in coelomocytes in response to injury. SpCoel1 is a single copy gene that is specifically expressed in most of the coelomocytes of the adult purple sea urchin, Strongylocentrotus purpuratus. The 4-kb transcript from this gene has a relatively short (426 nucleotide) open reading frame (ORF) with long 3' and 5' untranslated regions. The ORF encodes a protein that has strong amino acid sequence similarity to profilins from yeast to mammals. Transcript titrations of SpCoel1 show significant increases per coelomocyte in animals that have been physiologically challenged. Increases in transcript levels are of similar magnitudes between animals receiving different treatments, such as injuries from needle punctures or from injections of foreign cells. The evidence presented here implies a molecular mechanism by which this lower deuterostome defense system responds to external insult, viz that an external "injury signal" activates a signal transduction system, which in turn mediates the alterations in cytoskeletal state that are required for coelomocyte activation.


Alternative pathways for the sorting of soluble vacuolar proteins in yeast: a vps35 null mutant missorts and secretes only a subset of vacuolar hydrolases. vps35 mutants of Saccharomyces cerevisiae exhibit severe defects in the localization of carboxypeptidase Y, a soluble vacuolar hydrolase. We have cloned the wild-type VPS35 gene by complementation of the vacuolar protein sorting defect exhibited by the vps35-17 mutant. Sequence analysis revealed an open reading frame predicted to encode a protein of 937 amino acids that lacks any obvious hydrophobic domains. Subcellular fractionation studies indicated that 80% of Vps35p peripherally associates with a membranous particulate cell fraction. The association of Vps35p with this fraction appears to be saturable; when overproduced, the vast majority of Vps35p remains in a soluble fraction. Disruption of the VPS35 gene demonstrated that it is not essential for yeast cell growth. However, the null allele of VPS35 results in a differential defect in the sorting of vacuolar carboxypeptidase Y (CPY), proteinase A (PrA), proteinase B (PrB), and alkaline phosphatase (ALP). proCPY was quantitatively missorted and secreted by delta vps35 cells, whereas almost all of proPrA, proPrB, and proALP were retained within the cell and converted to their mature forms, indicating delivery to the vacuole. Based on these observations, we propose that alternative pathways exist for the sorting and/or delivery of proteins to the vacuole.


Actin- and tubulin-dependent functions during Saccharomyces cerevisiae mating projection formation. Several conditional-lethal mutant alleles of the single-copy Saccharomyces cerevisiae beta-tubulin and actin genes were used to evaluate the roles of microtubules and actin filaments in the pheromone-induced extension of mating projections. Mutants defective in tubulin assembly form projections indistinguishable in appearance from those formed by wild-type cells. However, the tubulin mutants are unable to move their nuclei into the projections and to orient the spindle pole body associated with each nucleus toward the projection tip. Actin mutants are defective in spatial orientation of cell-surface growth required for formation of normal mating projections. Migration of nuclei into mating projections and Spa2p segregation to projection tips are also defective in actin mutants. Studies with abp1 null mutants showed that the function of the Abp1p actin-binding protein is either not required for projection formation or there are other proteins in yeast with similar functions. Our findings demonstrate that actin is required to restrict cell-surface growth to a defined region for pheromone-induced morphogenesis and suggest that nuclear position and orientation in mating projections depend on direct or indirect interaction of microtubules with actin filaments.


Phosphorylation of FAR1 in response to alpha-factor: a possible requirement for cell-cycle arrest. Exposure of yeast a cells to alpha-factor causes cells to arrest in the G1 phase of the cell cycle. The FAR1 gene is required for this cell-cycle arrest; its product is necessary for the inhibition of a G1 cyclin, CLN2. Earlier work demonstrated that alpha-factor caused an increase in the transcription of FAR1 severalfold over a measurable basal level. We now show that transcriptional induction of FAR1 from a heterologous promoter is not sufficient to inhibit CLN2 in the absence of alpha-factor. We also show that FAR1 is phosphorylated in response to alpha-factor and propose that this phosphorylation may be required for FAR1 activity.


Membrane potential modulates release of tumor necrosis factor in lipopolysaccharide-stimulated mouse macrophages. Lipopolysaccharide (LPS)-mediated synthesis of macrophage gene products such as tumor necrosis factor (TNF) is controlled by different signaling pathways. We investigated intracellular free Ca2+ (Ca2+ic) and the membrane potential as early cellular responses to LPS and their role in the synthesis and release of TNF. In peritoneal macrophages and in the RAW 269 mouse macrophage cell line, LPS and its biologically active moiety lipid A stimulated TNF synthesis but exerted no significant effects on these early cellular responses using Fura-2/Indo-1 to measure Ca2+ic and bis-oxonol, as well as the patch-clamp technique to monitor membrane potential. In contrast, the platelet-activating factor transiently induced both an increase in Ca2+ic and cell membrane depolarization but no significant TNF release. Increased extracellular K+ concentrations or K(+)-channel blockers, such as quinine, tetraethylammonium, or barium chloride, inhibited the LPS-stimulated release of TNF alpha, as well as the accumulation of cell-associated TNF alpha as found by enzyme-linked immunosorbent assay analysis, but did not inhibit TNF alpha mRNA accumulation. Concentrations of quinine (greater than 125 microM) or of enhanced extracellular K+ (25-85 mM) required to inhibit TNF production both significantly depolarized macrophages. These results indicate a lack of ion transport activities as early cellular responses of macrophages to LPS but suggest an important regulatory role of the membrane potential on the posttranscriptional synthesis and release of TNF in macrophages.


tra-2 encodes a membrane protein and may mediate cell communication in the Caenorhabditis elegans sex determination pathway. The Caenorhabditis elegans sex-determining gene, tra-2, promotes female development in XX animals. In this paper we report the cDNA sequence corresponding to a 4.7 kb tra-2 mRNA and show that it is composed of 23 exons, is trans-spliced to SL2, and contains a perfect direct repeat in the 3' untranslated region. This mRNA is predicted to encode a 1475 amino acid protein, named pTra2A, that has a secretory signal and several potential membrane-spanning domains. The molecular analysis of tra-2 loss-of-function mutations supports our open reading frame identification and suggests that the carboxy-terminal domain is important for tra-2 activity. We propose that in XX animals the carboxy-terminal domain of pTra2A negatively regulates the downstream male promoting fem genes. In XO animals, tra-2 is negatively regulated by her-1, which acts cell nonautonomously. Because hydropathy predictions suggest that pTra2A is an integral membrane protein, pTra2A might act as a receptor for the her-1 protein. We propose that in XO animals, the her-1 protein promotes male development by binding and inactivating pTra2A. The role of cell communication in C. elegans sex determination might be to ensure unified sexual development throughout the animal. If so, then regulation of sexual fate by her-1 and tra-2 might provide a general model for the coordination of groups of cells to follow a single cell fate.


A histone variant, H2AvD, is essential in Drosophila melanogaster. H2AvD, a Drosophila melanogaster histone variant of the H2A.Z class, is encoded by a single copy gene in the 97CD region of the polytene chromosomes. Northern analysis shows that the transcript is expressed in adult females and is abundant throughout the first 12 h of embryogenesis but then decreases. The H2AvD protein is present at essentially constant levels in all developmental stages. Using D. melanogaster stocks with deletions in the 97CD region, we have localized the H2AvD gene to the 97D1-9 interval. A lethal mutation in this interval, l(3)810, exhibits a 311-base pair deletion in the H2AvD gene, which removes the second exon. P-element mediated transformation using a 4.1-kilobase fragment containing the H2AvD gene rescues the lethal phenotype. H2AvD is therefore both essential and continuously present, suggesting a requirement for its utilization, either to provide an alternative capability for nucleosome assembly or to generate an alternative nucleosome structure.


Sec23p and a novel 105-kDa protein function as a multimeric complex to promote vesicle budding and protein transport from the endoplasmic reticulum. A cell-free protein transport reaction has been used to monitor the purification of a functional form of the Sec23 protein, a SEC gene product required for the formation or stability of protein transport vesicles that bud from the endoplasmic reticulum (ER). Previously, we reported that Sec23p is an 84-kDa peripheral membrane protein that is released from a sedimentable fraction by vigorous mechanical agitation of yeast cells and is required for ER to Golgi transport assayed in vitro. We have purified soluble Sec23p by complementation of an in vitro ER to Golgi transport reaction reconstituted with components from sec23 mutant cells. Sec23p overproduced in yeast exists in two forms: a monomeric species and a species that behaves as a 250- to 300-kDa complex that contains Sec23p and a distinct 105-kDa polypeptide (p105). Sec23p purified from cells containing one SEC23 gene exists solely in the large multimeric form. A stable association between Sec23p and p105 is confirmed by cofractionation of the two proteins throughout the purification. p105 is a novel yeast protein involved in ER to Golgi transport. Like Sec23p, it is required for vesicle budding from the ER because p105 antiserum completely inhibits transport vesicle formation in vitro.


A novel transcriptional enhancer is involved in the prolactin- and extracellular matrix-dependent regulation of beta-casein gene expression. Lactogenic hormones and extracellular matrix (ECM) act synergistically to regulate beta-casein expression in culture. We have developed a functional subpopulation of the mouse mammary epithelial cell strain COMMA-1D (designated CID 9), which expresses high level of beta-casein, forms alveolar-like structures when plated onto the EHS tumor-derived matrix, and secretes beta-casein unidirectionally into a lumen. We have further shown that ECM- and prolactin-dependent regulations of beta-casein occur mainly at the transcriptional level and that 5' sequences play an important role in these regulations. To address the question of the nature of the DNA sequence requirements for such regulation, we analyzed the bovine beta-casein gene promoter in these cells. We now have located a 160-bp transcriptional enhancer (BCE1) within the 5' flanking region of the beta-casein gene. Using functional assays, we show that BCE1 contains responsive elements for prolactin- and ECM-dependent regulation. BCE1 placed upstream of a truncated and inactive beta-casein promoter (the shortest extending from -89 to +42 bp with regard to the transcription start site) reconstitutes a promoter even more potent than the intact promoter, which contains BCE1 in its normal context more than 1.5 kb upstream. This small fusion promoter also reconstitutes the normal pattern of regulation, including a requirement for both prolactin and ECM and a synergistic action of prolactin and hydrocortisone. By replacing the milk promoter with a heterologous viral promoter, we show that BCE1 participates in the prolactin- and ECM-mediated regulation.


Optimization of a model of full-thickness epidermal burns in the pig and immunohistochemical study of epidermodermal junction regeneration during burn healing. In order to obtain a wound model in which healing involved epidermis rebuilding and epidermodermal junction (EDJ) regeneration without involvement of any dermal repair, we optimized a previous model of experimental cutaneous burning with an aluminum bar by testing various conditions of burning associated with different pre- and postburn skin treatments. On the optimized model of full-thickness epidermal burns without any dermal injury, we investigated the kinetics of regeneration of 4 EDJ components, from day 2 to day 23 after burning. The epidermal healing was studied by light microscopy and EDJ regeneration by indirect immunofluorescence with one bullous pemphigoid (BP) serum, antisera to fibronectin and to type IV collagen (Coll IV) and the monoclonal antibody 4C 12-8 to laminin. Histologically, neoepidermis, detected from day 2, appeared as a reepidermization tongue which progressed from the burn edges between the overlying necrotic burned epidermis and the underlying uninjured dermis. Epidermis continuity was found to be restored at day 9. Immunohistochemically, labelling of BP antigen (BPA), Coll IV and laminin extended all along the neo-EDJ, from day 2 to day 23. In contrast, fibronectin labelling was detected only in the proximal and median portions of the neo-EDJ before day 7, then all along the neo-EDJ, from day 7 to day 23. For all the components except Coll IV, the intensity of the labelling beneath the neoepidermis was higher than that of the residual labelling remaining under the necrotic epidermis. Therefore, BPA and laminin regenerated synchronously to neoepidermis whereas fibronectin first regenerated with delay, then synchronously.(ABSTRACT TRUNCATED AT 250 WORDS)


Photochemotherapy with topical khellin and sunlight in vitiligo. In order to evaluate the efficacy of topical khellin the vitiligo macules of one side only were painted in 41 patients with a 2% solution of khellin in acetone and propylene glycol (90 and 10%, respectively) and exposed to sunlight for a period of 4 months with 3 weekly applications and with exposure times up to 90 min. The macules of the other side were treated in 36 of the 41 patients with acetone and propylene glycol only and sun-exposed with the same schedule, while in the remaining 5 patients they were neither treated with khellin or placebo nor sun-exposed. No significant difference was evidenced between the khellin and placebo-treated sides: no excellent result (repigmentation more than 75% of the affected area) was found, and good results (repigmentation more than 50%) were found in 24.9% of khellin- plus sunlight-treated macules and in 22.3% of placebo- plus sunlight-treated macules.


Telogen effluvium due to recombinant interferon alpha-2b. Five of 10 patients receiving long-term recombinant interferon alpha-2b therapy complained of considerable hair loss. The pull test and the trichogram showed a telogen count consistent with a telogen effluvium. In 2 patients the telogen count progressively improved despite continuing the treatment, whereas in 3 patients the trichogram continued to show an increased telogen count until interferon therapy was discontinued. Pohl-Pinkus hair constrictions were also present in 2 patients.


Alopecia areata and interleukin-2 receptor. Interleukin-2 (IL-2) is a lymphokine produced by activated T cells and its receptor (IL-2R) is expressed on T cells; one of the IL-2R components can be measured as a soluble serum protein (sIL-2R). Levels of sIL-2R were measured as a sign of T cell activation in serum of 12 patients suffering from alopecia areata (totalis or universalis) and in a group of healthy control subjects. An enzyme-linked immunosorbent assay was used to determine the levels of sIL-2R in blood samples drawn during both the active and resting phase of the disease. In patients with alopecia areata in active phase the sIL-2R concentrations were significantly higher than in stable phase and in controls. The biologic role of sIL-2R is still unknown, but one could speculate that T lymphocyte activation with subsequent secretion of IL-2 and IL-2R expression may contribute to the immune inflammatory mechanism of alopecia areata.


Cutaneous complications of artificial hair implantation: a pathological study. Five patients who developed severe cutaneous complications after artificial hair implantation were subjected to a scalp biopsy. The pathology showed the presence of hyperplastic epidermal proliferations that produced infundibulum-like structures around the implanted fibers in the superficial dermis. A dense acute inflammatory infiltrate surrounded the artificial fibers in the superficial dermis. In the deep dermis a granulomatous infiltrate was present whereas in the hypodermis the inflammatory infiltrate was sparse and the fibers were embedded in fibroplasia. The pathology of a patient who did not present any skin inflammation after artificial hair implantation showed similar pathological features but the absence of acute inflammation suggesting that bacterial infections play a major role in the development of the cutaneous complications of hair implantation. Since definitive treatment of the infections is ineffective until the fibers are removed from the scalp, surgical treatment was required in 2 of our patients.


Keratosis follicularis spinulosa decalvans: report of a case with ultrastructural study and unsuccessful trial of retinoids. Keratosis follicularis spinulosa decalvans (KFSD) is a genetic disorder characterized by disseminated follicular hyperkeratosis, especially localizated to scalp and face. We report the case of a new patient displaying typical features of KFSD. Ultrastructural study was performed and displayed round keratohyalin granules in follicular keratinocytes. Trial with etretinate, which has not been reported before in this disease, proved to be ineffective.


Wells' syndrome: report of 2 cases.

Wells' syndrome: report of 2 cases. Wells' syndrome is a rare dermatological disease whose origin remains uncertain. Based on clinical, biological and etiological data, it might be recognized as a major form of urticaria. The triggering factors are numerous and unspecific. As revealed by the 51Cr-EDTA test which measures intestinal permeability, the lesions do not seem to be caused by an excess of antigens passing through the intestinal mucosa.


Acute generalized exanthematic pustulosis induced by cefaclor and acetazolamide. We report a case of acute generalized exanthematic pustulosis induced by 2 different drugs, cefaclor and acetazolamide. The diagnosis was confirmed by challenge tests, and these 2 drugs respectively were proven to be responsible for the patient's pustular eruptions.


Systemic mastocytosis treated with histamine H1 and H2 receptor antagonists. A case of systemic mastocytosis with skin characteristics of telangiectasia macularis eruptiva perstans is reported. Systemic involvement was confirmed as a combination of highly increased urinary excretion of methyl imidazole acetic acid (Melm AA) and increased amounts of mast cells in skin, liver and colon transversum. Treatment with cimetidine (H2 receptor antagonist) resulted in diarrhea, however, the addition of cyproheptadine (H1 receptor antagonist) was beneficial in amelioration of the cutaneous symptoms of mastocytosis. The excretion of Melm AA was unchanged during treatment. After 7 months of continuous cimetidine and cyproheptadine treatment no side effects were observed.


Is Microsporum canis infection about to become a serious dermatological problem? In Europe, especially in the Mediterranean, the incidence of Microsporum canis infection has been on a steep increase during the recent years. In some countries (Italy) and geographic areas (Slovenia), M. canis is the most often isolated dermatophyte. In Slovenia (Yugoslavia), a dramatic increase in the incidence of M. canis infection has been observed recently, both in absolute figures and as compared to the rest of isolated dermatophytes. M. canis is a cause of tinea most prevalent in children. All attempts made during the past 6 years to eliminate the natural source of infection, represented mainly by stray cats, have failed. M. canis infection is becoming a serious epidemiologic problem in Europe. For its solution, integrated efforts of medical and veterinary services, and probably more stringent rules and controls in this particular area, will be required.


Evaluation of the frequency of contact allergic reactions to Kathon CG in the Maastricht area--1987-1990. In the period 1987-1990, 999 patients suspected of a contact allergy were patch-tested with Kathon CG 100 ppm active ingredients aq. Of these patients, 84 (8.4%) showed a positive reaction (70% females). In 62% of these cases clinical relevance was established. Most of them suffered from contact dermatitis of the face, the hands or the perianal region.


Sensitization to the isothiazolinone biocide. Report of the Swiss Contact Dermatitis Research Group 1988-1990. The rate of sensitization to isothiazolinones (Kathon CG) detected in Switzerland rose from 3.5% (out of 2,491 patients) in 1987 to 6.3% (out of 982 patients) in 1988 and 5.6% (out of 2,295 patients) in 1989-90. This rate of sensitization appears to be related to the more and more widespread use of isothiazolinone biocide in cosmetics, domestic products and in industry.


How to better understand the angry back syndrome. The angry back or excited skin syndrome consists of patch test reactions that are positive only when associated with adjacent or distant inflammatory skin lesions. Recent experiments in the mouse and man have provided the knowledge on the critical role of tumor necrosis factor in contact dermatitis as well as its importance for the induction of adhesion molecules. Based on these new findings we speculate on mechanisms that might explain this poorly understood clinical phenomenon.


Optical illusions in clinical dermatology: the Mach band phenomenon and afterimages. A survey of dermatologists was conducted to determine whether the perceptions of afterimages and Mach bands impacted on clinical dermatology practice. 26.5% (13/49) of respondents indicated that they perceived one or both of these optical illusions. No false-positive potassium hydroxide interpretations of skin scrapings for hyphae were reported due to perceived afterimages, and no skin biopsies were reportedly performed as a result of Mach bands.


Polymorphisms of HLA class III genes in allergic contact dermatitis. HLA class III polymorphisms (BF, C4A, C4B) were studied in 55 patients of different age and sex suffering from allergic contact dermatitis, with sensitization to different substances. In the overall group of patients no significant correlation between the disease and HLA markers was found. BF F allele was present in 34% and BS S in 64% of patients suffering from allergic contact dermatitis to nickel only versus 16.45% (relative risk, RR = 2.61) and 80.76% (RR = 0.42), respectively, of the control population. The BF FB subtype frequency was 23.91% versus 7.57% in the control samples (RR = 3.88). We thus hypothesize that this polymorphic serum protein might be involved in the pathogenesis of allergic contact dermatitis to nickel.


Contact allergy to isothiazolinone derivatives: unusual clinical presentations. Contact allergic reactions to the mixture of 5-chloro-2-methyl-4-isothiazoline-3-one and 2-methyl-4-isothiazoline-3-one are most frequently associated with intolerance to cosmetics. The present article points out that such reactions, particularly on the face, can have unusual clinical presentations that are very similar to seborrheic eczema, lupus erythematosus, lymphocytic infiltrate or photodermatitis. Atopic dermatitis is also often erroneously suspected.


Treatment of atopic dermatitis by allergen-antibody complexes: long-term clinical results and evolution of IgE antibodies. This study describes the long-term follow-up of the clinical response of 10 adult patients suffering from atopic dermatitis (AD) who were treated by administration of complexes made of Dermatophagoides pteronyssinus (Dpt) allergens and autologous antibodies specific to that allergen. We have already described the clinical improvement observed after 1 year of treatment involving regular injections of complexes; this improvement was maintained throughout a second year, even though the number of injections was greatly reduced. At the end of the second year of treatment, 5 patients were completely free of disease, and 3 had had a short-lasting recurrence of low-severity dermatitis. Using a disease intensity index the mean improvement for these 8 patients was 83% compared to baseline values. One patient showed a significant recurrence of symptoms, and 1 patient left the study for personal reasons when she was in good clinical condition. A significant reduction of specific anti-Dpt IgE antibody titers was observed in 7 out of the 8 patients in clinical remission, while the level of total IgE antibodies was unchanged until the very end of the study. This study not only confirms that clinical benefit can be obtained from the treatment of AD patients hypersensitive to Dpt by injections of allergen-antibody complexes but also indicates that the therapy induces a suppression of IgE antibody production that is specific for the particular allergen.


Pigmented dermatofibrosarcoma protuberans (Bednar tumour). A case of pigmented dermatofibrosarcoma protuberans in a 37-year-old man is reported. The tumour presented as a nodular lesion located in the left scapular area. Surgical resection with wide margins was performed. No recurrences have appeared to date. The presumptive origin of this entity is briefly discussed.


Vitiligo-like depigmentation and morpheas after specific intralymphatic immunotherapy for malignant melanoma. We report the case of a patient with stage 2 malignant melanoma (MM), who received a specific immunotherapy consisting of intralymphatic injections of irradiated MM cells. She developed subsequently vitiligo-like leukoderma and several plaques of localized scleroderma. Simultaneously an increase in the patient's cytotoxic activity against MM cells was detected in vitro. The responsibility of immune phenomena and specific immunotherapy for the appearance of depigmentation and morpheas is discussed.


Postoperative hyperemia of the glans penis with ulcers following revascularization surgery for vascular impotence. A case of postoperative hyperemia of the glans penis following revascularization surgery for erectile impotence is described. This new surgical concept (Hauri) combines arterial revascularization with an arteriovenous shunt. The so-called 'postoperative hyperemia of the glans penis' is observed in 10-20% of the cases after arterialization of the dorsal penile vein, i.e. a pure arteriovenous shunt, and in 2-3% of the cases in arterioarterial anastomoses. The ulcers of the glans penis, which occurred 1 year after the operation, healed under careful local anti-inflammatory and antibiotic therapy.


Focal epithelial hyperplasia.

Focal epithelial hyperplasia. In 2 children (brother and sister) focal epithelial hyperplasia could be diagnosed by means of histology, human papillomavirus typing and Southern blot. In the 14-year-old girl the history could be followed over many years. The high familial incidence and the special geographic distribution of focal epithelial hyperplasia are pointed out.


Congenital hypertrophy of the lateral nail folds of the hallux. A case of a 7-month-old boy with bilateral symmetrical congenital hypertrophy of the lateral nail folds of the hallux is described. The body showed no other skin symptoms. The types of ingrowing nails in infancy are discussed. A partial remission of the disease after 1 year was observed.


Localized unilateral hyperhidrosis.

Localized unilateral hyperhidrosis. We describe a case of localized unilateral hyperhidrosis on the forehead in a 52-year-old woman. The pathophysiology of this unique clinical picture and its treatment are described.


Targetoid hemosiderotic hemangioma.

Targetoid hemosiderotic hemangioma. A new case of hemosiderotic targetoid hemangioma is reported. The literature is briefly reviewed.


Mycetoma of the foot due to Fusarium sp. treated with oral ketoconazole. We report the case of a 30-year-old Nigerian patient with mycetoma of the foot without bone involvement caused by Fusarium sp..Long-term administration of ketoconazole produced a significant improvement with good clinical and biological tolerance.


Syringolymphoid hyperplasia with alopecia--a syringotropic cutaneous T-cell lymphoma? Report on a 44-year-old male patient who developed a solitary plaque on the trunk. Histologically there is hypertrophy of adnexal duct structures and a lymphoid infiltrate with a T helper phenotype. The term 'syringotropic variant of mycosis fungoides' is proposed for this distinct disorder.


Disseminated granuloma anulare: therapy with vitamin E topically. Topical use fo vitamin E (E-Mulsin) is recommended as an alternative treatment modality in otherwise recalcitrant granuloma anulare.


Fox-Fordyce disease: successful treatment with topical clindamycin in alcoholic propylene glycol solution. We report a 66-year-old woman with a 3-year history of intensely pruritic follicular papules in the axillar, pubic and inguinal areas. Previous treatment with topical fusidic acid and gentamicin sulfate was ineffective. The clinical and histological examination was consistent with Fox-Fordyce disease. Application of clindamycin in an alcoholic propylene glycol solution led to the clearing of the lesions within 1 month. Nine months later, the treatment was stopped, and no recurrence was observed.


Systemic corticosteroids do not prevent postherpetic neuralgia. We review the use of corticosteroids in preventing postherpetic neuralgia (PHN) in a retrospective study over 5 years and 10 months. Out of 113 patients evaluable, 46 (40%) had PHN. 21 of these 46 patients (38%) had received prednisone (p = 0.49; n.s.). Duration and intensity of PHN were not different in the prednisone-treated group. This long-term study does not support the use of prednisone for preventing PHN.


Histiocytic lymphophagocytic panniculitis (Rosai-Dorfman disease): a case report. A 68-year-old woman presented with a symmetrical enlargement of the face and submandibular area associated with essential thrombocytosis. Biopsy of the lesions revealed a septal and lobular panniculitis disclosing a lymphohistiocytic and plasma cell proliferation with lymphophagocytosis by histiocytes, compatible with Rosai-Dorfman disease. The lesions regressed spontaneously without therapy over a period of 4 months. Diagnosis was confirmed by electron-microscopic examination and immunocytochemical presence of S-100 antigen within the histiocytic cells. A 2-year follow-up did not demonstrate any evidence of recurrence.


From one to another.

From one to another. Organ transplantation is now well established as an excellent treatment for many previously fatal diseases of vital organs. For an organ to be of use to the recipient the cells must be alive but apart from kidney and hepatic lobe donation from a close blood relative, organs are removed from dead patients. The cause of death is usually a head injury or intracerebral bleed with permanent and complete destruction of the brain stem. Thus organ donation to save a life is usually associated with a tragedy for the donors and their families.


A cardiac transplant programme--the nurses role. In 1967 the first heart transplant was performed by Dr Christian Barnard in Capetown. Attempts to establish the technique at units worldwide produced poor results, mainly due to rejection and infection. In the 1970s, particularly at Stanford University Hospital, California, further work related to the detection and treatment of rejection allowed improvements in patient management and subsequent encouraging results in mortality and quality of life. The major breakthrough was, however, due to the introduction of cyclosporin into clinical practice in 1983. This immunosuppressive agent significantly reduced the mortality related to rejection. At the same time progress was made in several countries to establish the legal definition of brain death and to develop means of transporting donor organs over long distances.


Total quality management within the NHS. Total quality management is a system which enables managers to analyse and develop their services within a common framework. It incorporates the needs and preferences of service users in the setting and improvement of quality standards, within resource constraints.