Studies on antibody to intrinsic factor.

Sera from a group of 79 patients with pernicious anemia were studied for the presence of antibody to intrinsic factor. Two general types of antibody activity were found, and it was possible to distinguish three groups of pernicious anemia sera on the basis of their content of these types. Type I antibody blocks the binding of radioactive vitamin B(12) to intrinsic factor when added to intrinsic factor before the B(12); it is not detected on intrinsic factor when added after B(12). This antibody blocks intrinsic factor-mediated B(12) absorption in vivo when mixed in the sequence intrinsic factor + antibody I + B(12), but not when mixed in the sequence intrinsic factor + B(12) + antibody I. Type II antibody reacts with intrinsic factor when B(12) is attached. This antibody prevents the absorption of B(12) from intrinsic factor in pernicious anemia patients when mixed in the sequence intrinsic factor + B(12) + antibody II, and is thereby distinguished from antibody I.

Iron homeostasis in plasmacytoma-bearing mice.

Plasmacytomas in mice have been shown to contain significant quantities of nonheme iron, half of which is in the form of ferritin. The livers of animals with plasmacytomas have a decreased iron content, and plasma iron is reduced. The animals develop an anemia that is partially corrected by parenteral iron. However, when plasmacytoma-bearing mice are given excessive amounts of parenteral iron, the surplus is stored in the liver of animals rather than in the tumor itself. These findings suggest that plasmacytomas sequester iron and deprive other organs of it to satisfy their own need. However, the tumors do not function as a storage site for iron above their own need.

Expression of voltage-gated chloride channels in human glioma cells.

Voltage-gated chloride channels have recently been implicated as being important for cell proliferation and invasive cell migration of primary brain tumors cells. In the present study we provide several lines of evidence that glioma Cl- currents are primarily mediated by ClC-2 and ClC-3, two genes that belong to the ClC superfamily. Transcripts for ClC-2 thru ClC-7 were detected in a human glioma cell line by PCR, whereas only ClC-2, ClC-3, and ClC-5 protein could be identified by Western blot. Prominent ClC-2, -3, and -5 channel expression was also detected in acute patient biopsies from low- and high-grade malignant gliomas. Immunogold electron microscopic studies as well as digital confocal imaging localized a portion of these ClC channels to the plasma membrane. Whole-cell patch-clamp recordings show the presence of two pharmacologically and biophysically distinct Cl- currents that could be specifically reduced by 48 hr exposure of cells to channel-specific antisense oligonucleotides. ClC-3 antisense selectively and significantly reduced the expression of outwardly rectifying current with pronounced voltage-dependent inactivation. Such currents were sensitive to DIDS (200-500 microm) and 5-nitro-2-(3-phenylpropylamino) benzoic acid (165 microm). ClC-2 antisense significantly reduced expression of inwardly rectifying currents, which were potentiated by hyperpolarizing prepulses and inhibited by Cd2+ (200-500 microm). Currents that were mediated by ClC-5 could not be demonstrated. We suggest that ClC-2 and ClC-3 channels are specifically upregulated in glioma membranes and endow glioma cells with an enhanced ability to transport Cl-. This may in turn facilitate rapid changes in cell size and shape as cells divide or invade through tortuous extracellular brain spaces.

Aldose reductase, NADPH and NADP+ in normal, galactose-fed and diabetic rat lens.

NADPH and NADP+ levels were measured in rat lens from normal controls, from galactose-fed and diabetic rats during the first week of cataract formation. The level of NADPH in normal rat lens was determined to be 12.3 +/- 0.4 nmol/g wet weight, and that of NADP+ 4.6 +/- 0.2 nmol/g wet weight. In early cataract formation NADPH levels decreased rapidly during the first 2 days and then remained stable at 76% of control for galactose-fed and 84% for diabetic rats. NADP+ levels increased by 38% of control for galactose-fed and 54% for diabetic rats. Calculated NADPH/NADP+ ratios dropped from 3.36 +/- 0.21 to 1.86 +/- 0.16 in galactose fed rats, and from 2.81 +/- 0.15 to 1.61 +/- 0.16 in diabetic rats (P less than 0.001 for both experimental groups). These data are consistent with rapid NADPH oxidation during onset of lens cataracts. No significant changes in aldose reductase enzymatic activity levels were observed in either the galactosemic or the diabetic rats during the times measured.

Splenic sequestration with sickle cell-C disease.

During an episode of pneumonia and multiple pulmonary thromboembolic events, an adult male patient with sickle cell-C hemoglobinopathy developed a severe anemia associated with rapidly increasing splenic size and diminished splenic uptake of technetium Tc 99m sulfur colloid, indicating diminished splenic function due to hypersequestration, a rare and noteworthy phenomenon in adult patients with sickle cell-C disease.

Effectiveness of triglycyl vasopressin in persistent hematuria associated with sickle cell hemoglobin.

Two cases of persistent hematuria associated with the presence of sickle cell hemoglobin were treated intravenously with triglycyl vasopressin, a drug not previously used for this condition. One patient, a 16-year-old boy, had hemoglobin AS. Both patients had a history of severe hematuria persisting over several months, resistant to the usual forms of therapy, and requiring numerous transfusions. In each patient, the condition responded to intravenous triglycyl vasopression therapy, with cessation of hematuria. Experimental studies in dogs indicate that triglycyl vasopressin reduces renal blood flow substantially. Further trial of triglycyl vasopressin in severe hematuria associated with the presence of sickle cell hemoglobin appears to be indicated.

Complement and antibody mediate enhancement of HIV infection by increasing virus binding and provirus formation.

Previous studies have shown that infection of complement receptor (CR2)-bearing cells with HIV pretreated with antibody (Ab) plus complement (C) resulted in increased virus expression. The current study was designed to determine whether C-mediated 'enhancement' of HIV-1 production was the result of increased virus infection of cells as assessed by provirus formation and virus binding. Virus was incubated with anti-HIV Ab and/or C and added to CR2-positive MT-2 cells. Increased virus expression by MT-2 cells correlated with increased numbers of HIV-immunofluorescent-positive cells at 24 and 48 h and higher levels of provirus detected 8-28 h after infection. MT-2 cells also bound threefold more Ab-plus-C-treated virus than untreated virus. Serial dilutions of C showed that high levels of C with Ab did not enhance but rather suppressed virus expression. Studies were also performed which showed that terminal C components C5 and C8 were not necessary for the enhancing effect. The increased binding of C-coated HIV to CR-positive cells has important implications for the fate of virus in vivo.

Detection of HIV-1 provirus in bronchoalveolar lavage cells by polymerase chain reaction.

This study was undertaken to evaluate whether HIV-seropositive individuals harbor HIV provirus in cells obtained by bronchoalveolar lavage (BAL). BAL cells were obtained from 14 HIV-positive patients undergoing bronchoscopy for evaluation of acute pulmonary symptoms. Cells were fractionated into macrophage-enriched and lymphocyte-enriched populations. The quantity of HIV-1 proviral DNA in the unfractionated BAL cells and in each population of fractionated cells was determined following polymerase chain reaction (PCR) amplification. Detectable quantities (3-90 copies/100,000 cells) of HIV-1 proviral DNA were found in unfractionated BAL cells in 12 of 14 patients. In the other two patients, provirus was detected after a sevenfold enrichment of lymphocytes. Provirus was also detected in BAL macrophages from 8/14 patients although proviral content was significantly higher in the lymphocyte fraction (133 +/- 72 vs. 35 +/- 22 proviral copies, p = 0.03). No correlation was seen with the ability to detect provirus in lymphocyte- or macrophage-enriched fractions and clinical diagnosis (e.g., Pneumocystis carinii pneumonia). The data suggest that lymphocytes are the predominant cells that contain provirus found in the lungs, although macrophages may be infected in some patients.

The effect of estrogens on erythroid stem cells in polycythemic mice.

Plethoric mice treated with pharmacological doses of estradiol have decreased concentration of erythropoietin-responsive cells (ERC) in the marrow. We used the methylcellulose-culture system for growth of erythroid stem cells (CFU-E and BFU-E) to define more accurately these estrogen-induced changes. As an animal model we utilized plethoric mice given repeated injections of estradiol cypionate and found that at 14 days after the onset of treatment there was no significant change in the concentration of femoral CFU-E whereas there was a significant decrease of the BFU-E content. Both CFU-E and BFU-E increased progressively in the spleen over a 42-day period. Addition of estradiol directly to the cell-culture system showed no effect on CFU-E growth but induced a significant depression of BFU-E growth. This depression seemed to require the presence of adherent cells. It is our hypothesis that estrogens suppress only the early stages of erythroid proliferation and/or differentiation by a mechanism involving possibly the stromal (adherent) cells of the marrow microenvironment.

Identifying the promoter region of the human brain sodium channel subtype II gene (SCN2A).

Sodium channel genes are highly regulated. To begin analyzing the human brain sodium channel subtype II gene, SCN2A, at the transcriptional level, we mapped multiple transcriptional start sites within a 397 bp stretch of the 5'-UTR and -flanking region. When inserted into a basic luciferase reporter vector, this 397 bp region can promote luciferase expression in transiently transfected neuroblastoma cells, but not in non-neuronal cells. Thus, this study provides the initial description of a functional promoter in a human voltage-gated sodium channel gene.

Effect of plasma proteins and temperature on echogenicity of blood.

An explanation is proposed for the echogenicity to ultrasound scanning at 5 mHz and above of unclotted blood under conditions of stasis. In vitro experiments using blood from normal subjects and from patients with myeloma revealed that: 1) lysis of red cells prevented echogenicity, 2) echogenicity increased with increasing hematocrit, fibrinogen and other macromolecules, and temperature, and 3) blood from myeloma patients showed increased echogenicity and rouleau formation, a form of aggregation seen on peripheral smears. From these experiments it was concluded that red cell aggregation is a major cause of echogenicity of unclotted blood, requiring both intact red cells and conditions which are known to enhance red cell aggregation, such as the presence of macromolecules and increased temperature.

Simultaneous attainment of high electron and ion temperatures in discharges with internal transport barriers in ASDEX upgrade

Internal transport barriers have been demonstrated to exist also under conditions with T(e) approximately T(i) approximately 10 keV and predominant electron heating of the tokamak core region. Central electron cyclotron heating was added to neutral beam injection-heated ASDEX Upgrade discharges with a preexisting internal transport barrier, established through programmed current ramping leading to shear reversal. Compared to a reference internal transport barrier discharge without electron cyclotron resonance heating, the electron heat conductivity in the barrier region was found not to increase, in spite of a fivefold increase in electron heat flux, and also angular momentum and ion energy transport did not deteriorate.

Inferior vena cava thrombosis. Unusual presentation of testicular tumor.

We report an unusual case of testicular tumor presenting as thrombosis of the inferior vena cava. However, in contrast to a previous report, thrombosis in this patient was not caused by compression of the vena cava by neoplasm metastases. In our patient, we postulate that tumor invaded the vena cava and that thrombosis was exacerbated by a hypercoagulable state induced by the tumor.

Measurement of proteases in human subgingival dental plaque by fluorescence polarization.

Fluorescence polarization (FP) was examined as a rapid quantitative method to assay the proteases in subgingival plaque. Protease activity was measured by a decrease in FP at 0.5-min intervals over 5 min, using BODIPY-alpha-casein, a protein substrate. To quantitate activity, the least absolute deviation (LAD) slope for each assay was determined. Protease activity increased with the quantity of plaque (r=0.416, P<0.001). Of the 208 subgingival plaque samples, 87 contained detectable protease activity, with a mean of about 4 microg trypsin equivalents above a general background of 1 microg per site. The mean plaque protease activity of 89 paired samples from 15 individuals had decreased by 1.1 microg trypsin equivalents per site when measured at 8 months after tooth scaling and root planing (P<0.01). Most isolates of Porphyromonas gingivalis, Treponema denticola, Prevotella nigrescens, and Prevotella intermedia implicated in the pathogenesis of adult periodontitis exhibited high activity in the FP assay. The assay is rapid, quantitative and requires only one-tenth of the plaque sampled using a single pass with a Gracey curette at a single tooth site.

The effect of protein deprivation on erythroid stem cell growth.

The anemia of protein deprivation is due mainly to a decrease in erythropoietin (Ep) production. In this study we investigated the effect of protein deprivation on the number of erythroid stem cells (CFU-E, BFU-E). Mice given a protein-free diet for six days had significantly fewer CFU-E and BFU-E numbers per femur compared to mice given a diet with normal protein content. The decrease in BFU-E cannot be ascribed solely to decreased Ep production and occurs before changes in the femoral multipotential haematopoietic stem cells (CFU-S).

[Three infants with epidural hematoma]. / Drie zuigelingen met een epiduraal hematoom.

Three infants, two boys aged 10 months and one girl aged 5 months, developed drowsiness and emesis within a few hours of a fall (after which they had not lost consciousness). Radiological examination revealed an epidural haematoma which was treated with emergency decompression. The children subsequently made a good recovery. An epidural haematoma is a potentially life-threatening event. More than 20% of all cases of epidural haematoma occur in childhood. In contrast with distinct symptoms in most adults and older children, an epidural haematoma in infancy can have a minimum of symptoms. The open fontanelle means compression occurs less rapidly, but consequently blood loss can be greater, leading to acute anaemia, hypovolaemic shock and consumptive coagulopathy.

The microvasculature in the equine distal phalanx: implications for fracture healing.

OBJECTIVES: To describe the intra-osseous microvasculature of the distal phalanx of the equine forelimb with regard to its potential clinical relevance. METHODS: Eleven clinically normal equine forelimbs were used from six adult horses (range: 4 to 18 years old) euthanatized for reasons unrelated to lameness. In each limb the median artery was catheterized at the level of the carpus and India ink was injected under constant manual pressure. The limbs were frozen and 5 mm thick sections of the foot were cut in the sagittal, coronal, or transverse planes on a band saw. The sections were fixed in 10% formalin and cleared using a modified Spalteholz technique. Once cleared, the sections were photographed and the microvascular anatomy identified. RESULTS: The vascular injections revealed a rich intra-osseous microvascular supply of the distal phalanx originating from the medial and lateral palmar digital arteries. In addition, numerous smaller vessels from the terminal arch, formed by anastomosis of the medial and lateral palmar digital arteries, could be seen branching into the distal aspects of the distal phalanx. This distal portion of the distal phalanx appeared more densely vascularized than the proximal part in all specimens examined. CLINICAL SIGNIFICANCE: The increased vascularity demonstrated in the distal portion of the distal phalanx appears to correlate with improved fracture healing reported in this area. This may also explain why healing fractures which involve both the distal and proximal portions of the distal phalanx have been described as progressing from distal-to-proximal.