Oligosaccharides accumulated in the bovineβ-mannosidosis kidney

Summary The phenotype of bovine-mannosidosis (-mannosidase deficiency), recently identified in Salers cattle, is similar to the caprine form of the disease (Abbittet al., 1991). This investigation was designed to characterize accumulated kidney oligosaccharides in bovine-mannosidosis. Oligosaccharides were extracted from the kidney of an affected Salers calf and purified by chromatographic techniques. The amount of accumulating oligosaccharides in 1 g of wet tissue was about 21µmol. Structures of derivatized oligosaccharides were characterized by high-performance liquid chromatography, mass spectrometry, methylation analysis and sequential exoglycosidase digestions. The major accumulating oligosaccharides were Man1-4GlcNAc and Man1-4GlcNAc1-4GlcNAc. Oligosaccharides accumulating in minor amounts were Man1-4GlcNAc1-4Man1-4GlcNAc, Man1-6Man1-4GlcNAc1-4GlcNAc and Man1-4GlcNAc1-4Man1-4GlcNAc1-4GlcNAc. As in caprine-mannosidosis, oligosaccharides with terminal-mannose residues and cleaved as well as uncleaved chitobiose linkages were identified in bovine-mannosidosis kidney. The accumulating oligosaccharides in tissue were thus identical in bovine and caprine-mannosidosis; however, the source of the novel oligosaccharides remains to be determined.     

Heterophilic interactions between cell adhesion molecule L1 and α<Subscript>v</Subscript> β<Subscript>3</Subscript>-integrin induce HUVEC process extension <Emphasis Type="Italic">in vitro</Emphasis> and angiogenesis <Emphasis Type="Italic">in vivo</Emphasis>

Cell adhesion molecule L1 was implicated in angiogenic processes, tumor formation and metastasis. Here, we provide evidence that the sixth Ig-like domain of L1 (L1Ig6) interacts with _{v 3} to induce process extension of human umbilical vein endothelial cells (HUVECs) in vitro and angiogenesis in vivo. HUVECs formed network-like structures on full-length L1 or L1Ig6 substrates comparable to structures found on matrigel. In the presence of mab _{v 3} or cyclic RGD, apoptosis was induced. In fibrin matrices where L1Ig6 was covalently incorporated, HUVECs formed multicellular and hollow processes through interactions between cell-surface _{v 3} and RGD-sites of matrix-immobilized L1Ig6. No such processes were induced by L1Ig6 having non-functional RDG-sites, or in the presence of mab _{v 3} or cyclic RGD. In those matrices, increased apoptosis was found. Co-immunoprecipitation of L1 or L1Ig6 with _{v 3} suggests close interactions. Furthermore, L1Ig6 stimulated HUVECs showed increased tyrosine phosphorylation of _{v 3} and phosphorylation of MAP kinases (ERK1 and ERK2) but not AKT indicating specific activation of _v and _{v 3} followed by activation of downstream kinases. Application of L1Ig6-modified fibrin matrices on CAMs induced 50–60% increased _v and _v3 protein expression and in vivo angiogenesis indicated by ~50% increased mean vascular length density. The results demonstrate angiogenic potential of L1Ig6 involving ligation and activation of _v3     

Interferonγ and tumour necrosis factorα induce a synergistic antiproliferative response in human Ewing's sarcoma cells in vitro

Three human cell lines derived from Ewing's sarcoma (RM-82, VH-64, and WE-68) were investigated to establish the influence of recombinant human interferon (rhIFN) and tumour necrosis factor (rhTNF) on cell proliferation and survival and to characterize IFN and TNF receptor expression. Incorporation of [³H]thymidine into cells was inhibited by rhIFN after 24 h of incubation. Half-maximal inhibition was observed with 10–80 U/ml rhIFN. A maximal effect (50%–70% inhibition of cell proliferation) was achieved by treatment of cells with 250 U/ml rhIFN. The influence of rhTNF on proliferation was found to differ among cell lines and varied with the concentration and the duration of exposure of cells to this cytokine. In WE-68 and VH-64 cells [³H]thymidine incorporation was not affected by rhTNF up to 2000 U/ml after 96 h of incubation, where-as in RM-82 cells the incorporation was inhibited by 35% after 48 h of incubation with 100 U/ml rhTNF. However, all cell lines showed a synergistic antiproliferative response to the combination of rhIFN and rhTNF after 24 h of incubation. The human recombinant cytokines interleukin(IL)-1, IL-1, IL-2, IL-3, IL-4, IL-6 and granulocyte/macrophagecolony-stimulating factor, tested alone and in combination with rhIFN and rhTNF, had no influence on cell proliferation. Binding studies in the cell lines with¹²⁵I-rhIFN revealed a dissociation constant (K _d ) of 160–306 pM and approximately 8000–13500 receptors/cell. Binding experiments with¹²⁵I-rhTNF indicated 430–1250 receptors/cell withK _d ranging from 13 pM to 162 pM. These data indicate that, among various cytokines, only IFN and TNF are capable of potently reducing Ewing's sarcoma cell growth in vitro. Our data suggest that IFN alone or in combination with TNF may be useful in the design of novel strategies in Ewing's sarcoma therapy.     

A Three-dimensional Analysis of Blood Vessels in Bronchioloalveolar Carcinoma

The three-dimensional architecture of blood vessels within lung adenocarcinomas has not been well studied. In 19 cases with bronchioloalveolar carcinoma with central fibrosis, we three-dimensionally examined blood vessel architecture in 150 m thick sections stained with elastin staining and anti-CD34 antibody. We examined four regions: normal alveoli and three regions within the tumor including an area adjacent to the normal alveoli (external area), an area in which tumor cells were replacing epithelial cells (replacement area), and a central fibrotic area (fibrotic area). Elastin staining showed that elastic fibers formed the framework of the alveoli, and the alveolar structure shrank more strongly to the center of the tumor due to folding of alveolar walls invaded by adenocarcinoma cells. We also measured three vessel parameters in these four regions. The vessel diameters were 4.08±1.10 m, 3.95±1.02 m, 5.04±1.56 m, and 6.11±2.23 m, the circumferences of those vessels seen as complete circles were 43.11±12.78 m, 43.71±12.87 m, 95.21±39.32 m, and 126.77±54.65 m; the lengths between vessel bifurcations were 13.28±3.08 m, 13.47±4.58 m, 24.91±9.66 m, and 41.82±28.08 m in the normal alveoli, and the external, replacement, and fibrotic areas, respectively. Blood vessel architecture changed such that the vessels became larger and coarser towards the center of the tumor. Our three-dimensional analysis suggests continuous remodeling of alveolar capillaries rather than angiogenesis within bronchioloalveolar carcinoma.     

Compound heterozygosity for a β∘-thalassemia (frameshift codons 38/39; -C) and a nondeletional swiss type of HPFH (A→C at NT -110, Gγ) in a Czechoslovakian family

Summary We have analyzed the levels and composition of the fetal hemoglobin (Hb F) in several members of a Czechoslovakian family with a heterozygosity for a newly discovered -thalassemia (codons 38/39; -C), or for a newly detected nondeletional hereditary persistence of fetal hemoglobin (a form of Swiss-HPFH with an AC mutation at nucleotide –100 5 to the Cap site of G), or with a compound heterozygosity for these two conditions. The Hb F level in the -thalassemia heterozygotes averaged 0.3% with low G values ( 28%) and relatively high AT values ( 50%), that in the two Swiss-HPFH heterozygotes averaged 0.8% with 95% G, while that of the compound heterozygote was 3.1% with 95% G. The low Hb F levels were determined with a recently published cation exchange high-performance liquid chromatography (HPLC) procedure that is accurate at the 0.1%–0.2% Hb F level [3]. This method, together with a reversed-phase HPLC procedure, made it possible to detect this unusual type of nondeletional G-HPFH and provided the data indicating that the increased Hb F in the compound heterozygote was derived mainly from the chromosome with the HPFH determinant.This study was supported in part by USPHS Research Grant HLB-41544     

Atypical myeloblastic leukemia with differentiation into “Paraneutrophils”

Summary A case of a patient is presented, who suffered from acute leukemia, showing predominantly atypical myeloblasts in the bone marrow. In contrast, a high amount of atypical neutrophils were present in the peripheral blood, as well as there was a conspicuous number of monocyte-like intermediate forms between the myeloblasts and the atypical segmented neutrophils. The cytochemical pattern of the Neutrophils was highly atypical, the main part being devoid of sudanblack B-positive lipids, as well as of peroxidase and chloroacylesterase activity, which characteristically are present in large amounts in the neutrophil granulocytes. Moreover, atypical distribution of alkaline phosphatase and naphthylamidase activities were encountered in these cells. The intermediate or transitional cells exhibited a cytochemical pattern different from that of normal blood monocytes, but corresponding to that of the atypical neutrophils. Thus, the assumption of a direct maturation way from myeloblasts to paraneutrophils with mononuclear, monocyte-like intermediate stages was supported by the results of cytochemical investigations. In contrast to the elevated number of neutrophils in the peripheral blood (10 350/mm³) very rare normal neutrophil precursors with typical cytochemical pattern were encountered in the haematopoietic organs.

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Zusammenfassung Es wird über eine Patientin mit einer akuten myeloischen Leukämie berichtet, die im Knochenmark überwiegend atypische Myeloblasten aufwies. Im Gegensatz dazu fanden sich im peripheren Blut eine große Anzahl ebenfalls atypischer Neutrophiler sowie eine beträchtliche Zahl monozytenähnlicher Zellen, die wirals Zwischenformen zwischen den atypischen Myeloblasten und den pathologischen Neutrophilen auffassen. Die pathologischen Neutrophilen zeigen ein ungewöhnliches zytochemisches Muster, in dem der größte Teil dieser Zellen keine Sudanschwarz-B-positiven Lipide sowie kaum Peroxydase-Naphthol-AS-D-Cloroacetat-Esterase-Aktivität aufweist. Diese drei zytochemischen Reaktionen geben normalerweise eine starke positive Reaktion in den neutrophilen Granulozyten. Außerdem wurde eine atypische Werteinteilung sowohl der alkalischen Phosphatase als auch der Naphthylamidase festgestellt. Die monozytenähnlichen Zwischenformen unterscheiden sich deutlich in ihrem zytochemischen Muster von normalen Monozyten und gleichen den atypischen neutrophilen Zellen. Diese zytochemischen Befunde stützen die Ansicht, daß es bei der Patientin zu einer Ausreifung der Myeloblasten über ein monozytenähnliches Zwischenstadium zu atypischen neutrophilen Granulozyten gekommen ist. Trotz der hohen Zahl neutrophiler Granulozyten im peripheren Blut (10 350/mm³) wurden in den blutbildenden Organen nur wenig typische neutrophile Vorstufen mit einem normalen Enzymmuster festgestellt.

     

Role of Central Interleukin-1β in Gastrointestinal Motor Disturbances Induced by Lipopolysaccharide in Sheep

Cytokines are involved in the symptoms of theacute phase response induced by infectious diseases inhumans as well as in animals, and interleukin-1(IL-1 ) has a pivotal role in these changes. The role of central IL-1 in the gastrointestinalhypomotility and fever evoked by intravenousadministration of lipopolysaccharide (LPS) and themechanisms involved, were investigated in sheep as anexperimental model. LPS (0.1 g/kg, intravenously)induced gastrointestinal hypomotility and fever thatwere significantly reduced by priorintracerebroventricular administration of IL-1receptor antagonist protein (IL-1ra, 2 g/kg). The effects of LPS were mimickedby intracerebroventricular IL-1 (50 ng/kg),whereas IL-1 injected intravenously at the samedose only caused a slight and transient fever withoutmodifying the gastrointestinal motility. Priorintracerebroventricular administration of thecyclooxygenase inhibitor indomethacin (100 g/kg) butnot the corticotropin-releasing factor (CRF) receptorantagonist -helical CRF_9-41 (5 g/kg) blocked alleffects evoked by both LPS and IL-1. These resultssuggest that in sheep, LPS induces digestive motordisturbances through a central release of IL-1 andprostaglandins.     

Local and regional immunotherapy of cancer with interleukin 2

Conclusion The results obtained in preclinical systems as well as the first clinical trials suggest that local IL-2 immunotherapy may represent a novel approach to the treatment of some neoplasms. However, the experimental results should be confirmed and substantially extended before definitive conclusions can be drawn. We hope that the data and considerations discussed in this article will facilitate thoughts concerned with future clinical IL-2 trials and be instrumental in the optimization of IL-2 cancer immunotherapy.The Journal of Cancer Research and Clinical Oncology publishes in loose succession Editorials and Guest editorials on current and/or controversial problems in experimental and clinical oncology. These contributions represent exclusively the personal opinion of the author     

Protein kinase-ζ interacts with munc18c: role in GLUT4 trafficking

Aims/hypothesis Insulin-stimulated glucose transport requires a signalling cascade through kinases protein kinase (PK) C/ and PKB that leads to movement of GLUT4 vesicles to the plasma membrane. The aim of this study was to identify missing links between the upstream insulin-regulated kinases and the GLUT4 vesicle trafficking system.Materials and methods A yeast two-hybrid screen was conducted, using as bait full-length mouse munc18c, a protein known to be part of the GLUT4 vesicle trafficking machinery.Results The yeast two-hybrid screen identified PKC as a novel interactor with munc18c. Glutathione S transferase (GST) pull-downs with GST-tagged munc18c constructs confirmed the interaction, mapped a key region of munc18c that binds PKC to residues 295–338 and showed that the N-terminal region of PKC was required for the interaction. Endogenous munc18c was shown to associate with endogenous PKC in vivo in various cell types. Importantly, insulin stimulation increased the association by approximately three-fold. Moreover, disruption of PKC binding to munc18c by deletion of residues 295–338 of munc18c or deletion of the N-terminal region of PKC markedly inhibited the ability of insulin to stimulate glucose uptake or GLUT4 translocation.Conclusions/interpretation We have identified a physiological interaction between munc18c and PKC that is insulin-regulated. This establishes a link between a kinase (PKC) involved in the insulin signalling cascade and a known component of the GLUT4 vesicle trafficking pathway (munc18c). The results indicate that PKC regulates munc18c and suggest a model whereby insulin triggers the docking of PKC to munc18c, resulting in enhanced GLUT4 translocation to the plasma membrane.     

Heat shock protein induction in rat pancreatic islets by recombinant human interleukin 1β

Summary Interleukin 1, potentiated by tumour necrosis factor , is cytotoxic to pancreatic Beta cells in vitro. We have hypothesized that interleukin 1 induces oxygen free radicals in Beta cells. Since cytotoxicity induced by free radicals and by heat may activate the same cellular repair mechanism (the heat shock response), the aim of this study was to investigate the pattern of protein synthesis in isolated islets after exposure to interleukin 1 (150 pg/ml, 24 h), tumour necrosis factor (50 ng/ml, 24 h), heat shock (43°C, 30 min) and H₂O₂ (0.1 mmol/l, 20 min). By polyacrylamide gel electrophoresis, autoradiography, Western-blot analysis and partial peptide mapping of ³⁵S-methionine labelled islets, interleukin 1 was found to induce a 73 kilodalton protein belonging to the heat shock protein family heat shock protein 70, a heat shock protein 90, and haem oxygenase. A minor induction of heat shock protein 73 and haem oxygenase was seen after H₂O₂. Interleukin 1 did not induce heat shock proteins in rat thyroid cells, rat mesangial cells or in human monocytes. Tumour necrosis factor did not induce selective protein synthesis. Pre-exposure of islets to heat, tumour necrosis factor , or H₂O₂ did not prevent the impairment of glucose-stimulated insulin release seen after 24 h of interleukin 1 exposure. The data are compatible with free radical induction by interleukin 1. However, the heat shock response is not specific for oxidative injury, and previous studies have shown discrepant effects as to a protective effect of free radical scavengers against interleukin 1-mediated beta-cytotoxicity. Thus, a role for free radicals in this context is not definitely proven.     

B cell adrenoceptors and sulphonylurea-induced insulin release in mouse islets

Summary Interactions of tolbutamide and glibenclamide with B cell adrenoceptors have been reported. This study evaluated the possible role of such interactions in the stimulation of insulin release. Mouse islets were incubated in the presence of 10 mmol/l glucose alone or with tolbutamide (10 mol/l) or glibenclamide (0.02 mol/l). At 0.01–10 mol/l, blockers of ₂-adrenoceptors (yohimbine, idazoxan) or ₁-adrenoceptors (prazosin) had practically no effect on glucose-induced insulin release and did not affect its potentiation by sulphonylureas, except for a slight increase by 10 mol/l prazosin and idazoxan. Nonspecific -blockers (phentolamine, dihydroergotamine) increased control release at 10 mol/l, but only the latter amplified the response to tolbutamide. Blockers of -adrenoceptors were tested at 0.1–100 mol/l: propranolol (₁, ₂), metoprolol (₁) and compound ICI 118-551 (₂). They increased glucose-induced insulin release at 100 mol/l but variably altered the effect of sulphonylureas. Blockers of adrenoceptors have, thus, no effect on insulin release in vitro at therapeutic concentrations. At high concentrations, they non-specifically affect the action of sulphonylureas. We conclude that an interaction with B cell adrenoceptors is not involved in the insulinotropic action of sulphonylureas.     

“Bound” insulin: in vivo and in vitro biologic activity

Summary Partially purified preparations of human serum bound insulin were injected intraperitoneally or intravenously into intact fed, fasted or fasted-refed rats, or incubatedin vitro with their isolated epididymal adipose tissue. The biologic activity of bound insulin on muscle and adipose tissue,in vivo andin vitro, was compared with the activity of crystalline insulin standards. Bound and crystalline insulin injected intraperitoneally into rats stimulated the incorporation of glucose-u-¹⁴C into the glycogen of muscle and into the glycogen and fat of adipose tissue. The activities of both bound and crystalline insulin were affected by the nutritional state of the animals. Intravenous administration of partially purified bound insulin or crystalline insulin stimulated the incorporation of glucose-u-¹⁴C into the muscle glycogen of intact fed, fasted or fasted-refed rats and into the fat of the adipose tissue of fed or fasted-refed rats. Bound insulin, but not crystalline insulin stimulated fat synthesis in the adipose tissue of fasted rats. Bound or crystalline insulin, at the concentration used, failed to stimulate glycogen synthesis in adipose tissue. Bound insulin stimulated the oxidation of glucose into CO₂ in isolated rat adipose tissue and the incorporation of glucose-u-¹⁴C into the glycogen and fat of the adipose tissue. The effect of bound or crystalline insulin on isolated adipose tissue was also affected by the nutritional state of the animal.

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Zusammenfassung Teilweise gereinigte Präparate von gebundenem Insulin aus menschlichem Serum wurden normalen gefütterten, hungernden oder nach Hungern wiederaufgefütterten Ratten intraperitoneal oder intravenös injiziert oderin vitro mit dem isolierten epididymalen Fettgewebe dieser Ratten inkubiert. Die biologische Aktivität des gebundenen Insulins am Muskel und Fettgewebe,in vivo undin vitro, wurde mit der Aktivität von Standardlösungen kristallinen Insulins verglichen. Wurde gebundenes und kristallines Insulin Ratten intraperitoneal injiziert, kam es zu einer vermehrten Einlagerung von Glukose-U-¹⁴C in Muskelglykogen und in das Glykogen und Fett des Fettgewebes. Sowohl die Aktivität des gebundenen als auch des kristallinen Insulins wurden durch den Ernährungszustand der Tiere beeinflußt. Die intravenöse Verabreichung des teilweise gereinigten gebundenen Insulins oder des Kristallinsulins führte zu einer verstärkten Einlagerung von Glukose-U-¹⁴C in das Muskelglykogen der gefütterten, hungernden oder der nach Hungern wiederaufgefütterten Ratten und auch in das Fett des Fettgewebes der gefütterten und nach Hungern wiederaufgefütterten Ratten. Nur das gebundene Insulin, nicht das kristalline Insulin steigerte die Fettsynthese des Fettgewebes der hungernden Ratten. Weder das gebundene noch das kristalline Insulin führten bei den verwendeten Konzentrationen zu einer Steigerung der Glykogensynthese des Fettgewebes. Das gebundene Insulin stimulierte die Oxydation von Glukose zu CO₂ am isolierten Fettgewebe der Ratte und die Einlagerung von Glukose-U-¹⁴C in das Glykogen und Fett des Fettgewebes. Die Wirkung des gebundenen und des kristallinen Insulins auf das isolierte Fettgewebe wurde ebenfalls durch den Ernährungszustand der Tiere beeinflußt.

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Résumé Cette étude porte sur l'injection intrapéritonéale ou intraveineuse de préparations partiellement purifiées d'insuline liée de sérum humain. Des rats nourris ad libitum, mis à jeun, ou renourris après une période de jeûne, ont servi pour ces injections, et leur tissu adipeux épididymaire a servi à l'étude de l'effet in vitro de ces préparations. L'activité biologique de cette insuline liée a été comparée avec celle d'une insuline cristalline standard. L'insuline liée tout comme l'insuline cristalline stimule l'incorporation de glucose-u-C¹⁴ dans le glycogène du muscle et dansle glycogène et la graisse du tissu adipeux, à la suite de leur injection intrapéritonéale. L'état de nutrition des animaux injectés modifie les activités et de l'insuline liée et de l'insuline cristalline. L'injection intraveineuse d'insuline liée partiellement purifiée, ou d'insuline cristalline, stimule l'incorporation de glucoseu-C¹⁴ dans le glycogène musculaire de rats nourris ad libitum, à jeun, ou renourris après un jeûne prolongé. Elle stimule également l'incorporation du glucose dans la graisse du tissu adipeux de rats nourris ou renourris après jeûne. L'insuline liée seulement, non l'insuline cristalline, stimule la lipogénèse du tissu adipeux de rats à jeun. Tant l'insuline cristalline que l'insuline liée est restée sans effet, pour les dosages utilisés, sur la synthèse du glycogène dans le tissu adipeux. L'insuline liée stimule l'oxydation du glucose par le tissu adipeux isolé ainsi que son incorporation dans le glycogène et la graisse de ce tissu. L'action de l'insuline liée ou cristalline est également modifiée par l'état de nutrition de l'animal dont le tissu adipeux est utilisé in vitro.

     

Acute thermal injury of the esophagus

Acute thermal injury to the esophagus has not been reported previously in the radiographic literature. We present a case of a young adult who developed an intramura blister that ultimately communicated with the esophageal lumen. A double-contrast esophagogram outlined the resulting mucosal flap. A brief review of other injuries to the esophagus is included.     

Selective A2A adenosine agonist ATL-146e attenuates acute lethal liver injury in mice

Background d-Galactosamine (GalN)/lipopolysaccharide (LPS)-induced liver injury is an experimental model of fulminant hepatic failure in which tumor necrosis factor- (TNF-) plays a pivotal role. We examined the effects of a highly selective adenosine A_2A receptor agonist (ATL-146e) on GalN/LPS-induced fulminant hepatic failure.Methods Mice were given an intraperitoneal dose of GalN (800mg/g body weight)/LPS (100ng/g body weight) with and without ATL-146e (0.01µg/kg) treatment. Liver injury was assessed biochemically and histologically. Also, TNF- levels in the serum were determined.Results The serum liver enzyme (ALT) level in vehicle-treated mice was 20960 ± 2800IU/ml and was reduced by 63% to 7800 ± 1670IU/ml by treatment with 0.01µg/kg per minute ATL146e, P < 0.05. Treatment with ATL-146e significantly reduced serum TNF- and greatly reduced inflammation assessed by histopathologic examination compared with control mice treated with GalN/LPS. ATL-146e also reduced lethality at 12h from 65% to 13%.Conclusion The present findings suggest that the highly selective adenosine A_2A receptor agonist (ATL-146e) prevents endotoxin-induced lethal liver injury by suppression of TNF- secretion.     

Unspezifische Phosphatasen und Esterasen im Melanoblastom der Chorioidea

Zusammenfassung Melanoblastome der Chorioidea bestehen aus wenig pigmentierten Melanocyten und stark pigmentierten als Makrophagen bezeichneten perivasculären Zellen. In sieben histochemisch untersuchten Melanoblastomen der Chorioidea besaßen die Makrophagen eine starke Aktivität unspezifischer Esterasen und saurer Phosphatasen, in den Melanocyten war die Aktivität schwach. Ein Melanoblastom vom epitheloiden Typ enthielt vorwiegend Zellen mit starker Enzymaktivität und zeigt außerdem alle Übergänge zwischen Melanocyten und Makrophagen. In allen Melanoblastomen waren die Fraktionen der isodynamen Esterasen und sauren Phosphatasen (Zymogramme nach Agarelektrophorese des Tumorextraktes) gleichartig, lediglich die Aktivität der einzelnen Fraktionen ist etwas verschieden. Die als Makrophagen bezeichneten Zellen sind ebenso wie die Melanocyten tumoreigene Zellen. Die starke Phosphataseaktivität wird auf den verstärkten Einbau von Phosphat (P³²) in die Tumorzellen bezogen.

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Summary The malignant melanoma of the choroid consists of melanocytes with slight pigmentation and of tumor cells (macrophage-like) that are heavily pigmented and perivascularly localized. In 7 malignant melanomas of the choroid belonging to the spindle cell and epithelioid type (Reese), the macrophage-like tumor cells histochemically showed a high activity of nonspecific esterases and acid phosphatases; the activity in melanocytes was low. A malignant melanoma of the epithelioid type chiefly contained tumor cells with high enzymatic activity and showed all transitional cells from melanocytes to macrophages. Zymograms of all melanomas (electrophoresis of tumor extract on agar-agar) showed similar fractions of isodynamic esterases and acid phosphatases which differed only in activity. The macrophage-like cells and the melanocytes were tumor cells. A relation presumably existed between the high activity of phosphatases and the increased incorporation of phosphate (P³²) into the tumor cells.

     

Rheumatoid arthritis associated with transient gamma 2-heavy chain disease

Summary This paper describes the clinical history of a patient (F.-O.) with longstanding rheumatoid arthritis (RA), who subsequently developed a transient gamma 2-heavy chain disease (₂-HCD). Immunochemical studies comprised serial determinations of serum levels of intact IgG, the -HCD protein, IgA, and IgM. New applications of the rocket immunoselection and the radial immunodiffusion were used for the quantitation of the -HCD protein and intact IgG, respectively, in the presence of one another. Immunofluorescent microscopy on bone marrow cells showed cells containing -heavy chains but devoid of light chains. Protein studies of the isolated -HCD protein revealed a molecular weight of 72 000 in the dimeric form, a carbohydrate content of 9.7%, and a PCA-Val-Gln NH₂-terminal amino acid sequence. The literature on the rare coexistence of RA and -HCD in a single patient is reviewed.     

Dysphagia and expiratory air flow

Expiratory air flow preserves the freedom of the upper airway from foodway contamination in patients with dysphagia. Valving the tracheostomy cannula, quad coughing, the Heimlich maneuver, the supraglottic swallow, and coupling to a ventilator each has a place among the measures used for treating aspiration.     

Distribution pattern of α and β myosin in normal and diseased human ventricular myocardium

Summary All fibers in three normal, four dilated, and two ischemic human ventricles were classified according to their myosin content using three sets of monoclonal antibodies each specific for one myosin heavy chain isoform (, and ). Numerous fibers contained only myosin heavy chain (denoted as fibers), others contained either and , or and myosin heavy chain (denoted as and fibers, respectively). The percentages of fibers were systematically determined along the walls of seven homologous regions of the ventricular myocardium.In all ventricles, there was an -fiber transmural gradient, with less fiber in the subendocardium than in the subepicardium. More fibers were found in the right than in the left ventricular wall but there was no difference between the mid-portion and the apex of the free wall of each ventricle. The diseased ventricles contained a lower fiber percentage than the normal hearts. fibers were very rare in the normal ventricles (less than 5%) and almost inexistent in pathological hearts. The correlation between the mean fiber percentages of the diseased hearts and their cardiac indices (r=0.88, P<0.05) suggests that the small amount of myosin distributed in a large number of ventricular fibers could play a role in the contractile performance of the heart. In conclusion, this study provides evidence for 1) an fiber transmural gradient, and 2) a lower myosin ratio in discased than in normal human ventricle.This work was supported in part by L'Institut National de la Santé et de la Recherche Médicale 101 rue de Tolbiac, 75013 Paris     

Improvement in Obstructive Sleep Apnea in the Supine “Knees-Up” Position

We report a 38-year-old man with obstructive sleep apnea whose sleep-disordered breathing was substantially reduced by sleep in the supine, knees-up position, relative to his sleep in the customary supine, knees-down position. No obvious anatomic or pathophysiologic alterations explained this phenomenon. The effect was reproducible in the patient 4 years later. Potential mechanisms underlying such improvement, including alterations in upper airway/lung volume dependence and venous supply to upper airway vasculature, are discussed. This manipulation could be an important adjunctive treatment for a subset of obstructive sleep apnea patients demonstrating such an effect.     

Thalassemia intermedia: compound heterozygous β∘/β+-thalassemia and co-inherited heterozygous α+-thalassemia

Summary The relative excess of - over -globin chains in the erythroid precursors is the chief pathophysiological factor of homozygous -thalassemia. The clinical picture is usually characterized by a transfusion-dependent dyserythropoietic anemia (thalassemia major). However, some patients present with moderate anemia that does not require regular blood transfusions (thalassemia intermedia). The molecular heterogeneity of -thalassemia mutations and changes of - and -globin gene expression play an important role in modifying the clinical phenotype. We report here on a female Greek patient with homozygous -thalassemia but normal growth and development, excellent exercise tolerance, and no need of blood transfusions. She is thus mildly affected clinically, although there is marked pallor, jaundice, and hepatosplenomegaly. These signs correspond to her marked hypochromic, microcytic anemia with erythroid hyperplasia of the bone marrow. -Globin genotyping shows her to be compound heterozygous for the codon 39 C T -nonsense mutation and for the T C ⁺-mutation at position 6 of the splice consensus at the exon 1/intron 1 junction (CD39 C T/IVS 1–6 T C). -Globin gene mapping demonstrates the presence of a 3.7-kb ⁺-thalassemia deletion on one allele (–^3.7/). Taken together, this study identifies a complex interaction of genetic factors that do not significantly alter the clinical phenotype when present alone but ameliorate the course of homozygous -thalassemia when inherited in combination.Abbreviations Hb hemoglobin - Hct hematocrit - HPFH hereditary persistence of fetal hemoglobin - IVS intervening sequence - MCH mean corpuscular hemoglobin - MCV mean corpuscular volume - PCR polymerase chain reaction