[Development of specific immunotherapy for cancer]. We developed CTL therapy against cancer as an cancer specific immunotherapy. The primary CTL therapy showed certain efficacy on advanced cancer patients. However, when ratio of monocyte population in peripheral blood lymphoid cells (PBL) of advanced cancer patients increases more than that of CD8+ T cell population, so that CD8+ T cell/M3 ratio is less than 1, CTL directed to cancer cannot be induced in PBL and the primary CTL therapy is not materialized for these patients. Monocyte-macrophages in these patients release a suppressive factor(s) capable of inhibiting CTL induction and proliferation at induction phase but not inhibiting cytotoxic activity in target cell lysis at effector phase. Furthermore, these monocytes expressed strong message of TGF beta. If most of increased monocytes of PBL from advanced cancer patients are removed by a nylon wool column method from the PBL, CTL directed to cancer cells can be induced even in PBL of advanced cancer patients with high frequency, this improved CTL therapy against cancer showed remarkable efficacy as the cancer specific immunotherapy in various advanced cancer patients without any side effects.


[Adoptive immunotherapy using immobilized anti-CD3 mAb-activated autologous lymphocytes: the strong cytotoxicity was supported by CD16+ cells which proliferated in prolonged cultures]. Inflammatory cytokines are able to facilitate the homing of transferred lymphocytes, tumor cell lysis through induction of adhesion molecules, also able to reduce tumor cell susceptibility to LAK cells by increasing tumor cell class I antigen. Investigation with 12 cell lines suggested that promotion of lysis by ICAM-1 was more responsible than protection by (allogeneic) class I Ags (Fig. 1). PBMC were cultured in anti-CD3 coated flasks with rIL-2. CD3+ cells dominated until day 7, decreased thereafter with CD4+. CD8+ and CD16+ increased (Fig. 2). Strong cytotoxicity obtained in some cultures correlated well with CD16+, contributing exclusively among several variables to the activity estimation in multiple regression analysis (Fig. 4). Among 6 cases, in which 2 or more cycles of transfer was done, 1 was prophylaxis of recurrence, in 2 of 3 advanced metastasis cases in which cells were transferred as BRM in the course of chemotherapy, survival of half a year was obtained in good QOL with suppressed disease and adequate level of PBL number. In 2 other cases, inflammation eliciting local treatments were combined. In the case 4, three large liver metastasis from colon cancer which resisted topical ethanol injection and chemotherapy, responded to the transfer with reduced lesions to 1/8 (Fig. 8). In the case 5, abdominal metastasis from colon cancer were removed, liver metastasis were injected of ethanol, and cells were transferred. Responses were obtained to immunotherapy in a certain degree, while never to any chemotherapy.


[Establishment of EGF, EGF-R and FN-R positive human adenocarcinoma cell line (GAC-1)]. A human adenocarcinoma cell line designated as GAC-1, was established from ascites of the 56-year old male patient with rapidly progressive gastric cancer. The doubling time was about 18.5 hours in vitro, and cell cycle analysis using flow cytometry showed marked increase of S phase (46.1%). Immunohistochemical demonstration of GAC-1 cells revealed positive staining of TGF-alpha, EGF, EGF-R, FN-R, laminin and negative staining of fibronectin. Histogram of them indicated aneuploidy with modal number 57 and they formed tumors in nude mice.


[Development of an experimental model for spontaneous lymph node metastasis of human esophageal carcinoma in nude mice--histopathological analysis]. Three human squamous cell carcinoma cell lines (HPL-EsC-1-K, -S, and -M) originated from a male patient with esophageal carcinoma were established and were studied on their tumorigenic and metastatic properties in nude mice. All cell lines grew in the hind foot pads following subcutaneous inoculation and produced popliteal lymph node metastasis dose (2-8 x 10(6)/mouse)-dependently. Based on the histopathological findings on serial sections of the lymph nodes, the stages of lymph node invasion by cancer cells were classified into 4 stages (St. 0-III). The time course of lymph node metastasis of EsC-K cells were examined. Advanced stage of metastasis increased according to the time elapsed after tumor cell inoculation. Incidence of metastasis of EsC-K cells were not affected by host factors such as sex differences, anti-asialo GM1 antibody treatment on the hosts. Today, there are few experimental models for studies on spontaneous lymph node metastasis of human carcinomas. This experimental model provides a useful research tool for studies on the biology and therapy for lymph node metastasis of esophageal cancer.


[Establishment and characterization of a human squamous cell carcinoma cell line LK-52 which produce direct activator of coagulant factor X]. Uterus origin squamous cell carcinoma cell line LK-52 was established from surgical specimen of lung metastatic nodule. LK-52 produce poorly differentiated squamous carcinoma in nude mouse, and doubling time in vitro was 38 hours. Chromosome analysis show various abnormality and main mode number was 67 and 68. LK-52 shed active procoagulant substance into culture medium. The culture medium of LK-52 shortening recalcification time of normal human plasma and factor VII or factor IX deficient plasma but not factor X deficient plasma. Procoagulant activity of LK-52 product may induced with direct activation of coagulant factor X. Procoagulant activity which produced by cancer cell may play a important roll in the unbalanced haemostasis of cancer patient.


[Use of 2-mercaptoethanol in cell culture]. Survival and growth in in vitro cultivation of lymphocytes, lymphoma cells and some other cells including human carcinomas are profoundly improved by 2-mercaptoethanol. These cells hardly take up cystine, an essential nutrient in the culture medium, but in the presence of 2-mercaptoethanol they can utilize cystine. Recently it has been found that 2-mercaptoethanol is effective in the in vitro cultivation of pathogenic trypanosomes and in the in vitro development of bovine embryos. The mechanisms by which 2-mercaptoethanol improves the survival and growth of these cells are described.


Automated site-directed drug design: a method for the generation of general three-dimensional molecular graphs. A new algorithm for creating diverse, irregular and physically reasonable three-dimensional linear atomic chains is described. The linear chains of atoms, or molecular graphs, are generated by solving a series of trigonometric equations within geometric constraints for a given set of atom types. The nature and number of the chains that are produced can be controlled by changing the palette of atom types, so that a chemist user could generate template suggestions that are synthetically relevant to a drug design project. Testing has shown that the method is sufficiently robust to be used in a general context. The molecular graphs could serve as useful structural templates for joining up regions in an active site where a ligand might interact strongly with the receptor. This paper is concerned with the description and proof of the methodology. The approach will form part of a larger structural tool kit for helping chemists to design novel ligands for a specified site.


Automation of conformational analysis and other molecular modeling calculations. A software system has been developed for facilitating modeling calculations on large numbers of molecules. Using the system, it is possible to subject one or more molecules to a series of calculations, each requiring use of a different computer program. No user intervention is required: where necessary, output from one program is used automatically as input to the next. Names are assigned to output files automatically and in a systematic manner. As an example, the system can be used to perform a succession of calculations aimed at identifying the major low-energy conformers of each of a set of molecules, starting only from their chemical connectivities. The reliability of the results has been tested by calculations on 40 molecules taken from the Cambridge Structural Database. The observed crystal structure geometry could be found for the majority of these molecules.


Object Command Language: a formalism to build molecular models and to analyze structural parameters in macromolecules, with applications to nucleic acids. We have written a programming language OCL (Object Command Language) to solve, in a general way, two recurring problems that arise during the construction of molecular models and during the geometrical characterization of macromolecules: how to move precisely and reproducibly any part of a molecular model in any user-defined local reference axes; and how to calculate standard or user-defined structural parameters that characterize the complex geometries of any macromolecule. OCL endows the user with three main capabilities: the definition of subsets of the macromolecule, called objects in OCL, with a formalism from elementary set theory or lexical analysis; the definition of sequences of elementary geometrical operations, called procedures in OCL, enabling one to build arbitrary three-dimensional (3D) orthonormal reference frames, to be associated with previously defined objects; and the transmission of these definitions to programs that allow one to display, to modify and to analyze interactively the molecular structure, or to programs that perform energy minimizations or molecular dynamics. Several applications to nucleic acids are presented.


Finding and filling protein cavities using cellular logic operations. A method for solid-filling protein cavities is presented. The method uses a pattern-recognition technique based on cellular logic operations to distinguish between convex and concave regions of a protein. In doing this it solid fills protein cavities and automatically defines a boundary between cavity and exterior free space. The operations used to fill the cavities also can be used to process the filler to filter out small-scale features. So far the main use of the method has been in visualizing protein active sites for docking. The method can be used to find cavities of a given size range and could be used to find novel protein binding sites.


FOCUS: a program for analyzing molecular dynamics simulations, featuring digital signal-processing techniques. FOCUS is a program for analyzing molecular dynamics simulations. It enables the researcher to monitor structural and energetic properties during the trajectory, and to calculate the corresponding statistical averages, correlation functions and Fourier transforms. In addition to these conventional methods, the program also utilizes novel methods based on digital signal-processing techniques to characterize the various motions. The characteristic frequencies in the system are revealed by the frequency distribution function g(v), which is calculated from the Fourier transform of the atomic coordinates. A filtering technique is employed to remove uninteresting motion (e.g., high-frequency bond stretching) while retaining and focusing on important motion (e.g., low-frequency conformational motion). The filtering technique enables fast display of slow events without getting a blurry or jittery picture due to the high-frequency motions. Another new way for analyzing the motion is by extracting "characteristic modes" and associated frequencies. This yields a pictorial description of the oscillatory motions in a manner analogous to normal mode analysis.


XELE--a polypeptide model-building program for a graphics workstation. A model-building program, XELE, for use in protein crystallography has been written in C under UNIX on a graphics workstation. This program makes full use of the X Window system to display the electron density distribution and to manipulate the polypeptide model, and therefore is named XELE. It utilizes a fast three-dimensional rendering package, Dorè, and is portable to other types of graphics workstations. A part of the program for the man-machine interface uses the library of X Window and X Toolkit, and therefore is highly interactive. The structure analysis program package, PROTEIN, is also implemented in an interactive mode using X Window, and has been interfaced with XELE.


Improving data acquisition parameters of 31P in vivo spectra for signal analysis in the time domain. To obtain reliable NMR quantitation, experimental cautions concerning data acquisition must be taken when using automatic predictive calculations. For this study, 2000 31P in vitro and in vivo spectra were processed, using the enhancement procedure with linear prediction using singular value decomposition (EPLPSVD) method, and analyzed. The effects of quadrature detection modes (simultaneous or sequential), of the number of time-domain samples used are investigated and experimental conditions such as sample motions and spectral width are discussed.


Lanthanide complexes of aminophosphonates as shift reagents for 7Li and 23Na NMR studies in biological systems. A systematic NMR characterization of various Dy(III) complexes of linear and macrocyclic aminophosphonates as 7Li and 23Na NMR shift reagents for biological systems was undertaken. Their efficacy as shift reagents (SR) was tested under constant aqueous solution ionic strength conditions at pH 7.5 as a function of rho = [SR]/[M+]. Further characterization of the two best SRs, Dy(PcPcP)2(7-) and Dy(DOTP)5-, led to the conclusion that, although quite sensitive to solution pH and the presence of alkali metal ions and Mg2+ and Ca2+, these complexes were stable towards hydrolysis by phosphatases. The lack of precipitation of its solutions in the presence of Ca2+, allowed the choice of Dy(DOTP)5- as the best overall SR for biological studies. Other SRs, like Dy(TTHA)3-, although less sensitive to pH and to divalent ions, require significantly higher concentrations to yield the same shifts, leading to large bulk susceptibility artifacts in perfused tissues and organs.


Magnetic resonance spectroscopy: a powerful tool for drug metabolism studies. Studies on the metabolism and disposition of drugs using nuclear magnetic resonance spectroscopy (MRS) as the analytical technique are reviewed. An overview of the main studies classed in terms of the observed magnetic nucleus (1H, 2H, 7Li, 13C, 19F, 31P, 77Se) is followed by some typical examples of the way in which 19F and 31P MRS can be profitably employed to gain more understanding about the metabolism and disposition of the anticancer fluoropyrimidines (5-fluorouracil (FU) and its prodrugs) and ifosfamide (IF). The results of three recent studies carried out in our laboratory are developed. They concern the direct quantitative monitoring of the hepatic metabolism of FU in the isolated perfused mouse liver, the elucidation of the origin of the cardiotoxicity of FU and the metabolism of IF from an analysis of biofluids of patients. Finally, the advantages and limitations of MRS for investigations on drug metabolism are discussed.


High resolution NMR spectroscopy of physiological fluids: from metabolism to physiology. High resolution NMR spectroscopy of physiological fluids provides quantitative, qualitative and dynamic information on the metabolic status of the interstitial and plasma compartments under a variety of pathophysiological conditions. The simultaneous detection and quantitation by NMR spectroscopy of numerous compounds of the intermediary metabolism offers a new insight in the understanding of the milieu intérieur. NMR spectroscopy of physiological fluids offers a unique way to define and monitor the global metabolic homeostasis in humans. The development of this analytical approach is still limited by the scarcity of pluridisciplinary teams able to fully exploit the wealth of information present on the NMR spectrum of a fluid. While application in pharmacology and toxicology is already established, the main areas of current development are cancer, hereditary metabolic disorders, organ transplantation and neurological diseases.


Accurate estimation of inter-atomic distances in large proteins by NMR. Recently a method was proposed which permits the extraction of the exact interatomic distance information from the measurement of the evolution of a single cross-peak relative to the mixing time in a NOESY experiment. This is performed through a careful multi-exponential analysis allowing the extraction of the relaxation parameter, and, consequently, the inter-proton distance. We investigate in the present paper whether this technique, already evaluated theoretically, can be used in a real experimental case. We have recorded and analyzed a set of 56 NOESY experiments on a lysozyme sample. Some 81 nOe build-up curves obtained from these data were analyzed in terms of distance. It is shown that the correlation between the measured distances and the reference distances obtained from crystallographic studies, is quite good. An accuracy of the order of 10% is obtained.


Modelling the dynamics of an antigenic peptide using NMR relaxation data. The internal dynamics of a cyclic peptide which was designed to mimic an antigenic loop of the haemagglutinin, is studied through heteronuclear relaxation along the 13C alpha-1H alpha vectors and through homonuclear relaxation along the 1H alpha-1HN and 1H beta-1H beta' vectors. Order parameters are extracted from the longitudinal and cross-relaxation data. Molecular dynamics simulations are performed and the order parameters are calculated in different ways from the trajectories. The simulation, which is performed in vacuo, gives smaller order parameters (vector motions of larger amplitude) than the experimental results. However, the general features of the experimental order parameters are reproduced by the molecular dynamics simulation. The flexibility of the molecule can then be investigated from the results of the molecular dynamics. It shows that the mobility observed through the order parameters is due to motions in flanking regions, remote from the observed vectors.


[Discovery of carnosine and anserine. Some of their properties]. The history of discovery of carnosine and anserine is reviewed with special reference to the structure and distribution of the dipeptides in various tissues during ontogenesis. The state of the dipeptides in muscle cells, their metabolism and role in muscle activity are considered. The properties of carnosine and anserine phosphoric esters are described, and their putative role in mitochondrial oxidative phosphorylation is discussed. The membranotropic activity of carnosine and anserine is demonstrated.


Progress in multidimensional NMR investigations of peptide and protein 3-D structures in solution. From structure to functional aspects. 2-D and 3-D NMR techniques were used to investigate the conformations in solution of several peptides and proteins for which crystalline structures are not available yet. Insect defensin A is a small (40 aa) antibiotic protein exhibiting a characteristic 'loop-helix-beta-sheet' structure. A striking analogy was found with charybdotoxin, a scorpion toxin in which a CSH (cysteine stabilized alpha-helix) motif is also present. Wheat phospholipid transfer protein (PLTP) (90 aa) has a 3-D structure resulting from the packing of four helices and of a C-terminal less well-defined fragment. Preliminary results show that PLTP forms a complex with lyso-PC and that such an interaction results in a conformational change affecting principally the C-terminal half of the protein. A last example is given with surfactin, a lipopeptide biosurfactant from bacterial origin. Its protonated form shows a very compact structure in which the two acidic residues located on the top of a 'horse saddle' topology face each other, whereas the ionized form could adopt a more extended conformation. A common property of these compounds is their capacity to interact with lipids. The present structural data open the way for a further establishment of structure-activity relationships.


NMR characterization of a diamagnetic model of unliganded alpha chains from human hemoglobin. This paper reports the reconstitution and spectroscopic characterization of a complex between alpha globin from human adult hemoglobin and protoporphyrin IX-Zn(II). Optical and proton one-dimensional (1-D) NMR spectra indicate that the prosthetic group binds in a 1:1 stoichiometry to the apoglobin in a single conformation. Using 2-D proton NMR techniques we assigned resonances corresponding to the majority of porphyrin substituents and to several side chains of amino acids in contact with the porphyrin. Analysis of nuclear Overhauser enhancement interactions between identified protons indicated that the complex contains only one rotation isomer of the prosthetic group. The diamagnetic Zn(II) ion is coordinated to the proximal histidine (His87) and does not bind O2 or CO as a sixth ligand. The ring current effects on protons from the distal valine (Val62) are considerably higher than in the liganded form providing strong evidence for a more compact ligand binding pocket relative to the carbon monoxy state. Therefore, protoporphyrin-Zn(II)/alpha globin complex is a suitable diamagnetic model for unliganded alpha chains and will be used for structure determination by NMR and modeling methods.


[The place of carnosine among physiologically active peptides]. The results of analysis of EROP-Moscow data base concerning structural and functional peculiarities of endogenous regulatory oligopeptides are reviewed in relation to carnosine, the first endogenous peptide bioregulator. The dipeptide fragment ala-his is widely distributed in natural systems, in particular in various representatives of living organisms. The main structural peculiarity of carnosine is its elongated "filamentous" structure with a positively charged N-terminus and a cyclic radical characteristic of large physiologically active oligopeptides. The relatedness of carnosine to other oligopeptides also becomes apparent during the analysis of its role in various regulatory systems of the body.


[Carnosine: biological role and prospects for use in medicine]. The biological role of the histidine-containing dipeptide carnosine (beta-alanyl-L-histidine) has been reviewed. The properties and putative biological role of the dipeptide in vertebrate tissues are considered. The antioxidative activity of carnosine and related compounds is described. The author's conception of the membranoprotective effect of carnosine on cells, tissues, and whole organism has been formulated. The properties of carnosine as an antistressory radioprotective agent are discussed. The data presented suggest that carnosine is a perspective immunomodulating tool which has many applications in medicine.


The inhibition of bovine heart hexokinase by 2-deoxy-D-glucose-6-phosphate: characterization by 31P NMR and metabolic implications. The glucose analog, 2-deoxy-D-glucose (2DG), has been used widely for studying the initial steps in the metabolism of glucose by radio-isotope tracer methods and by 31P NMR. In the rat heart perfused with acetate/2DG (both 5 mM) plus insulin, trapping of phosphorus by 2-deoxy-D-glucose-6-phosphate (2DG6P) results in a steady state exhibiting high 2DG6P (55 mM) and low ATP concentrations but near-normal function, as observed in an earlier 31P NMR study. In order to understand how the 2DG6P concentration is stabilized, we studied the inhibition of a mammalian hexokinase by 2DG6P in vitro by a 31P NMR technique. Inhibition, previously unobserved, was found. It is similar to inhibition by G6P in that it is competitive with ATP and not competitive with 2DG, but the inhibition constant (1.4 mM) is much larger. The experimental protocol includes provisions for enzymatic destruction of stray inhibitors such as G6P. The results show that the high 2DG6P and low ATP concentrations found in the steady state of the perfused heart should strongly reduce the rate of phosphorylation of sugars by hexokinase.


NMR studies of interactions between inhibitors and porcine pancreatic phospholipase A2. Two-dimensional NMR studies were performed on the complexes of porcine pancreatic phospholipase A2, bound to a micellar lipid-water interface of fully deuterated dodecylphosphocholine, with competitive inhibitors derived from the following general structure: [formula: see text] X and Y are alkyl chains with various 'reporter groups'. The interactions between the inhibitor and the enzyme were localized by comparison of 2-D nuclear Overhauser effect spectra using protonated and selectively deuterated inhibitors, and inhibitors with groups having easily identifiable chemical shifts. These experiments led us to the following conclusions for the phospholipase A2/inhibitor/micelle complex: i) the His48 C2 ring proton is in close proximity to both the amide proton and the methylene protons at the sn-1 position of the glycerol skeleton of the inhibitor, ii) the acyl chain of the inhibitor at the sn-2 position makes hydrophobic contacts near Phe5, Ile9, Phe22 and Phe106; iii) no interactions between the acyl chain at the sn-1 position and the protein could be identified. Comparison of our results on the enzyme/inhibitor/micelle ternary complex with the crystal structure of the enzyme-inhibitor complex shows that the mode of inhibitor binding is similar. However, in several cases we found indications that the hydrophobic chains of the inhibitors can have multiple conformations.


[Carnosine and anserine as specialized pH-buffers--hydrogen ion carriers]. Data are reviewed that carnosine (pK 6.9) and anserine (pK 7.1) act as specific mobile pH buffers promoting a rapid elimination of pH gradients between different parts of muscle fibers (very large cells containing various structural components, such as actomyosin, Z-discs, mitochondrial and sarcoplasmic reticulum, etc.). The specific role of these muscle dipeptides as pH buffers (H(+)-translocators) provides explanation for the so-called "Severin phenomenon"-a manyfold enhancement by carnosine of the ability of the isolated muscle to contract and accumulate lactate.


[Carnosine and anserine in working muscles--study using proton NMR spectroscopy]. NMR spectroscopy was used to study carnosine and anserine metabolism in rat tissues under intensive muscle loading. Muscle loading was accompanied by the dipeptide (predominantly anserine) accumulation in muscle tissues. Preliminary per os administration of carnosine (250 mg/kg of body mass) did not increase the dipeptide content in muscle tissues but diminished the lactate content in rat muscles under intensive muscle loading.


13C NMR studies of bacterial fermentations. We describe the experimental methods used and the constraints that apply in studies of anaerobic cell metabolism by 13C NMR. We review some of the results of our recent work in this area. Clostridium neopropionicum was shown to ferment ethanol into propionate by the acrylate, non-randomizing pathway. The same metabolic route accounts for 50% of the propionate formed in the complex ecosystem that inhabits the pig's large intestine. The rest is formed via the randomizing succinate pathway. Reductive, hydrogenotrophic acetogenesis was studied in several ecosystems. Although it is usually overshadowed by methanogenesis in the competition for hydrogen, it may become an efficient electron sink when methane biosynthesis is blocked by a specific inhibitor.


[Use of gamma-aminobutyric acid from various sources in the synthesis of homocarnosine in the brain of animals of various ages]. The dynamics and intensity of radiolabelled carbon incorporation from [14C] putrescine into homocarnosine in the brains of rats of various age have been studied. In the brains of 1-, 7-, and 2-day-old rats putrescine is the main GABA source for homocarnosine synthesis. In the brains of 14- and 31-day-old animals the GABA formed from glutamic acid participates in homocarnosine synthesis alongside with putrescine. In the brains of adult rats the rate of [14C] incorporation from glutamic acid into homocarnosine is 7 times as low as that of [14C] incorporation from putrescine.


In vivo 31P NMR study of early cellular responses to hyperosmotic shock in cultured glioma cells. Cell volume regulation in the face of osmotic stress is a fundamental homeostatic activity, and is most critical in brain, which is spatially constrained. Despite the importance of this phenomenon, little is known about volume regulation in the brain, primarily because of the cellular heterogeneity in the tissue. We describe here simultaneous in vivo 31P nuclear magnetic resonance (NMR) measurements of cell volume, intracellular pH and phosphate metabolites during early responses to hyperosmotic stress in C6 glioma cells perfused in NMR-compatible bioreactors. Cell volume was measured using dimethyl methylphosphonate (DMMP) as a probe which has an intracellular NMR resonance shifted upfield from the extracellular resonance. The sensitivity of these measurements allowed 31P NMR spectra to be collected every 30 s. Following an increase in osmolarity from 320 to 480 mOsm by addition of NaCl to the perfusate, C6 glioma cells shrank to 67% of their original volume. We also observed a simultaneous increase of intracellular pH coincident with the decrease in cell volume. The signals from ATP decreased by 10%, but those from phosphocreatine (PCr) increased by 31% after hyperosmotic shock. However, correcting the ATP signals for the decrease in cell volume indicated that its intracellular concentrations increased after treatment. Signals from glycerophosphorylcholine (GPC) and glycerophosphorylethanolamine (GPE) were not changed significantly. This is the first in vivo report of early cellular responses monitored by NMR spectroscopy following hyperosmotic shock in cultured cells.


[Carnosine as a stimulator of cytotoxic and phagocytic function of peritoneal macrophages]. Biochemical changes in peritoneal macrophages and their relatedness to the cytostatic and phagocytotic function in C3HA mice injected with a single intraperitoneal dose of 0.45 mM carnosine and 4-methyluracil or stimulated with peptone have been studied. During the first 24 hours after injection both carnosine and 4-methyluracil increase the activity of adenosine deaminase and purine nucleoside phosphorylase, the key enzymes of purine catabolism which is the main source of O2-. radicals in macrophages. In carnosine-stimulated macrophages the activity of membrane 5'-AMP nucleotidase decreases on days 1-3 after injection which points to alleviation of adenosine-induced inhibition as well as to macrophage activation. Carnosine increases the cytostatic and phagocytotic activities of macrophage coupled to O2-. production. The mechanism of the stimulating effect of carnosine on macrophages seems to consist in the dipeptide interaction with specific receptors localized on the plasma membrane of macrophagal cells.


[Effect of carnosine and 4-methyluracil on the development of experimental hepatitis in rats]. A comparative study of the hepatoprotective effect of carnosine and 4-methyluracil under CCl4-induced acute toxic hepatitis has been carried out. The extent of liver injury and its regeneration were established from morphological data as well as from changes in the activities of alanine aminotransferase (ALT) and histidase and the bilirubin content in blood serum. Hyperlipoperoxidation in the liver and serum was assessed by the amount of TBA-active products. It was found that by day 10 of experimental hepatitis ALT and histidase levels in blood sera of untreated animals exceeded the normal values 1.3- and 3.9-fold, whereas those in the carnosine-treated group approximated the values characteristic of intact animals. The activity of serum ALT in animals treated with vitamin B12 or 4-methyluracil exceeded normal values 1.5 and 1.6 times, whereas that of histidase was 2.5 and 2.7 times as high. Carnosine and 4-methyluracil inhibited (in approximately the same degree) the formation of TBA-active products in the liver. According to morphological dta, cessation of CCl4 injections was accompanied by rapid regeneration of liver tissues in all animal groups. Carnosine enhanced regenerative processes in parenchymatous and connective tissues in a far greater degree in comparison with other drugs. The mitotic index in the carnosine-treated group exceeded more than twofold the corresponding parameters in untreated animals. Possible mechanisms of carnosine action on liver repair are discussed.


[The effect of carnosine on the liver enzyme system in the irradiated body]. The effect of carnosine on post-radioactive changes in lipid peroxidation (LPO) products in blood serum and cytochrome P-450 content in liver microsomes has been studied. Per os administration of carnosine 24 hours prior to irradiation in a minimal lethal dose (7 Gr) markedly decreases the post-radioactive accumulation of LPO products in rat blood serum one hour after irradiation and fully restores the post-radioactive decrease in the cytochrome P-450 content in rat liver microsomes on day 5 after irradiation. Besides, the ability of carnosine to prevent the post-radioactive decline in the activity of UDP-glucuronyl transferase. Another key enzyme of the liver detoxifying system, has been demonstrated. The data obtained testify to the ability of carnosine to provide effective protection against post-radioactive intensification of LPO in irradiated organisms.


[The effect of carnosine on hematopoietic stem cell activity in irradiated animals]. Carnosine given to adult animals together with potable water one day prior to gamma-irradiation or injected in a single intraperitoneal dose one hour after irradiation enhances colony formation by haemopoietic stem cells migrating from the bone marrow to the spleen. In young animals with a high colony-forming activity carnosine either decreases or does not influence at all the efficiency of colony production.


[The effect of autoblood in a hyposmotic state on the colony-forming activity of hematopoietic stem cells]. It has been shown that hypoosmotic autoblood injected sub- or intracutaneously stimulates the colony-forming activity of haemopoietic stem cells in mice. Autoblood injected to animals immediately after their irradiation stimulates haemopoiesis even after a single dose. When mice are injected with autoblood prior to irradiation, the time between the first injection and the day of irradiation is critical for manifestation of the immunomodulating effect. Autoblood infusions immediately before, the day before, or two days before irradiation markedly deteriorate the clinical status of experimental animals and cause death in some of them. It is suggested that stimulation of haemopoiesis is associated with the appearance in the blood stream of a population of radiosensitive cells, apparently T-cell precursors.


[The protective effect of carnosine in hypoxia and reoxygenation of the isolated rat heart]. The effect of carnosine (15 mM) on the contractile activity of isolated rat hearts contracting in an isotonic regime (37 degrees C at a 5 Hz stimulation frequency) has been studied. Carnosine added to the perfusing solution had no effect on the contractile activity either in hypoxia or during reoxygenation but decreased it with a simultaneous increase in the coronary flow during reoxygenation. Carnosine inhibited by 60% the lactate dehydrogenase release from cardiac cells. A conclusion is drawn that the protective effect of carnosine is due to its membrane-stabilizing action which is implemented during inhibition of peroxidation of membrane lipids.


[Reperfusion injury of myocardial biomembranes after acute fatal hemorrhage and their correction with carnosine]. Studies on inbred male rats revealed that ischemia associated with clinical death caused by haemorrhage with subsequent recirculation and reoxygenation during resuscitation led to an increase in enzyme activities in blood serum and perfusate passed through the coronary bed of isolated hearts, presumably due to impaired membrane integrity. Damage to cardiomyocyte membranes was caused by intensification of LPO concomitant with enhanced glycolysis and lactate accumulation in the cardiac muscle. Carnosine (25 mg/kg) injected simultaneously with recirculation and reoxygenation prevented membrane damage and created optimal conditions for the functioning of membrane enzymes.


[Prospects for designing medicines based on carnosine (several new applications)]. The perspectives in application of carnosine, its analogs (histidine-containing dipeptides), and their derivatives as components of medicinal drugs are reviewed. These applications are based on antioxidative properties of carnosine and its analogs, their chelating activity towards transient valency metals as well as on their specific neurotransmitter functions in the brain. Combination of carnosine with other antioxidants and the use of copper or zinc complexes with histidine-containing dipeptides are considered as perspective trends in the design of new drugs.


A VLA-4 alpha-chain specific monoclonal antibody enhances CD3-induced IL-2/IL-2 receptor-dependent T-cell proliferation. Cross-linking the T-cell receptor-associated CD3 complex using the immobilized monoclonal antibody OKT3 can induce low levels of proliferation of purified resting T cells. The effect of coimmobilizing a monoclonal antibody 19H8 specific for the alpha-chain of the integrin VLA-4 on T-cell activation was evaluated. The level of proliferation induced by coimmobilization of the anti-VLA-4 with OKT3 was about 2- to 3-fold over proliferation induced by maximal OKT3 stimulation. The costimulatory activity of 19H8 was dependent on CD3 stimulation since immobilized 19H8 by itself did not induce proliferation. IL-2 secretion was found to be increased over 2-fold with 19H8 costimulation. Addition of exogenous IL-2 resulted in enhanced proliferation of both OKT3 and OKT3 plus 19H8-stimulated cells, but T cells coactivated with 19H8 exhibited a greater capacity to proliferate in response to exogenously supplied IL-2. Analysis of IL-2 receptor expression by flow cytometry revealed that the percentage of CD25-positive cells activated with either OKT3 or OKT3 plus 19H8 is comparable, but the mean fluorescence of cells coactivated with 19H8 is about 3-fold over cells stimulated with OKT3 alone. Dependency of the 19H8 enhanced proliferation on the IL-2/IL-2 receptor system was established by using IL-2-specific neutralizing antisera that reduced the proliferation of T cells activated with OKT3 alone or OKT3 plus 19H8 to comparable levels.2+hese results demonstrate that adhesion molecules may operate at the level of cytokine production and expression of its receptors to modulate the activation state of a cell.


Ex vivo secretion of tumor necrosis factor and interleukin-2 by rat splenocytes after intestinal ischemia and shock. We studied the ex vivo secretion of tumor necrosis factor (TNF) and interleukin 2 (IL-2) by splenocytes after circulatory shock induced by intestinal ischemia and reperfusion in rats. Shock was induced by total occlusion of the superior mesenteric artery followed by reperfusion. In a second group, vascular occlusion was maintained throughout the experimental protocol. A third group of sham rats and a fourth group of control rats with a negligible surgical procedure were also studied. "Spontaneous" (untriggered) secretion of TNF by splenocytes was higher in the ischemia-reperfusion group than in all other groups (p < 0.01), but did not increase significantly after stimulation with LPS. Splenocytes from control rats exhibited a marked increase in TNF secretion after stimulation with LPS to values similar to those in the ischemia-reperfusion group. A diminished, though statistically significant increase in LPS-stimulated secretion of TNF was detected in the sham and ischemia only groups of rats (p < 0.05) from untriggered values in each. Untriggered secretion of IL-2 was similar in all groups. However, when compared to control rats, splenocytes from the three surgically manipulated groups exhibited suppressed secretion of IL-2 in response to stimulation with Con A (p < 0.05). These results support the role played by TNF in mediation of shock and point to spleen macrophages as a source of TNF after intestinal ischemia and reperfusion. Our results also demonstrated postinjury alteration in immune function manifested by depressed ability of splenocytes to increase the production of IL-2 after stimulation with Con A.


Transforming growth factor-beta 1 down-regulates expression of membrane-associated lymphotoxin and secretion of soluble lymphotoxin by human lymphokine-activated killer T cells in vitro. Lymphokine-activated T killer lymphocytes (T-LAK) are important effector cells in various diseases of tissue destructive reactions. They require stimulation with various cytokines to proliferate and mature into function effector cells. We have examined the role of various endogenously and exogenously added cytokines, and tumor necrosis factor (TNF)/lymphotoxin (LT) receptors in this process in vitro. The present report is a continuation of these studies. We found that human T-LAK cells express membrane-associated LT (mLT) but not TNF, and secrete high amount of soluble LT (sLT) but low levels of TNF. When added to the initial cultures or immature T-LAK cells, transforming growth factor-beta 1 (TGF-beta 1) suppressed both mLT expression and sLT secretion by 30-40%. Coculture of mature T-LAK cells with TGF-beta 1 caused 35% down-regulation of both mLT expression and sLT secretion after 18 h of incubation. Kinetic experiments indicated reduction of LT mRNA synthesis could occur in as little as 1 h when cocultured with 5 ng/ml of TGF-beta 1. TGF-beta 1 also reduced mLT induced T-LAK cell cytolytic activity on L929 cells in vitro. It appears TGF-beta 1 can down-regulate LT mRNA syntheses, mLT expression, and sLT secretion of human T-LAK cells in vitro.


Effects of tumor necrosis factor alpha (TNF-alpha) on collagen arthritis. Louvain rats were administered tumor necrosis factor alpha (TNF-alpha) via a continuous osmotic infusion pump. These rats were then immunized with native type II collagen (CII) to determine the effects of exogenous TNF-alpha on collagen-induced arthritis (CIA), an animal model of rheumatoid arthritis (RA). In this highly susceptible strain, 100% of experimental and control rats developed arthritis although TNF-alpha-treated rats had more severe disease as judged by both clinical and blinded radiographic parameters. Humoral responses to collagen were high in both groups, but cellular responses to CII were augmented by TNF-alpha. Serum IL-6 levels were significantly increased in all arthritic rats. This study suggests that TNF-alpha is a proinflammatory cytokine in CIA and that future studies targeting TNF-alpha might be therapeutic.


Development and applications of a radioimmunoassay (RIA) for the in vitro and in vivo quantification of murine IL-1 beta. A specific, precise, and accurate radioimmunoassay (RIA) for murine interleukin-1 beta (mIL-1 beta), with a sensitivity of 250 pg/ml, has been established. Although mIL-1 beta shares structural homology and multiple biological properties with mIL-1 alpha, this RIA did not detect mIL-1 alpha or other murine cytokines such as TNF and IL-6. Recombinant mIL-1 beta, freshly added in different concentrations to murine plasma, was recovered from 88 to 104%, and intra- and interassay coefficients of variation never exceeded the 10% value. Parallel analysis showed that murine plasma and cell or organ supernatants did not affect the test. This characteristic allowed mIL-1 beta analysis directly in the nonmanipulated biological specimens. In murine macrophage supernatants collected after 24 h of in vitro stimulation with LPS, nanogram fractions of IL-1 beta were detected by RIA. These values corresponded to approximately 50% of the total IL-1 detected by the LAF bioassay. In spleen, liver, and lung, IL-1 beta appeared at significant levels (110 ng/g of lung, 638 ng/g of spleen, and 78 ng/g of liver) as early as 1 h after LPS administration, reached the plateau 1-2 h later, and then slowly but progressively decreased. In plasma and brain, nanogram fractions of IL-1 beta were detectable by 4 h post-LPS. Thereafter, IL-1 beta levels progressively increased to reach the value of 44 ng/g in the brain and 2 ng/ml in plasma 8 h after LPS treatment.


Overexpression of metallothionein confers resistance to the cytotoxic effect of TNF with cadmium in MCF-7 breast carcinoma cells. Experiments were designed to determine whether cells that overexpress metallothionein acquire resistance to the cytotoxic action of tumor necrosis factor (TNF). Human MCF-7 mammary carcinoma cells, which are sensitive to the cytotoxic action of TNF, were stably transfected with a vector in which the human metallothionein-IIA (hMT-IIA) gene was placed under the control of the constitutively active beta-actin promoter. MT-expressing clones displayed greater resistance to cadmium toxicity than control cell lines. Neither control-transfected nor MT-expressing cell lines were sensitive to the cytotoxic effect of TNF alone, even at concentrations as high as 200 ng/ml. However, treatment of control-transfected cells with TNF in the presence of CdCl2 produced a greater cytotoxic effect than CdCl2 alone. MT-expressing cell clones were protected from this synergistic cytotoxic effect of TNF and CdCl2. These results suggest that under certain conditions MT expression may protect tumor cells from the cytotoxic effects of TNF.


Prothymosin alpha enhances human natural killer cell cytotoxicity: role in mediating signals for NK activity. We have investigated the effects of prothymosin alpha (ProT alpha) on the in vitro NK activity of various human cell populations obtained by successive purification of large granular lymphocytes (LGLs) and CD16+ cells. The results of this study indicate that ProT alpha is able to enhance the spontaneous NK activity of cells from normal donors. This effect was time and dose dependent in the range 7 to 350 nM, occurred over a wide range of effector/target cell ratios, and appeared not to require the presence of accessory cells. In addition, ProT alpha exhibited additive effects with recombinant interferon-gamma (rIFN-gamma) and recombinant interleukin-2 (rIL-2). Finally our data suggest that the effect of ProT alpha enhancing the NK cell cytotoxicity appears to involve enhancement of p70 IL-2R expression and internalization of IL-2, and was independent of cell proliferation.


Growth velocity of some fetal parameters. I. Brain weight and brain dimensions. In this study, fetal growth velocities of the brain weight (whole brain and infratentorial part) and of the right and left fronto-occipital diameters were established from 420 normal fetuses and neonates. The age of the subjects ranged from 10 to 41 gestational weeks. Growth rates were computed by time intervals, and velocity curves were plotted with their 95% confidence intervals. The whole brain displays an accelerating pattern of growth until 35 weeks; thereafter, the growth rate breaks down. The growth velocity of the infratentorial part of the brain increases throughout fetal life, without a fall at 35 weeks. Moreover, it grows faster than the whole brain after 24 weeks. The lengths of the hemispheres present a steady decreasing pattern of growth velocity throughout pregnancy.


Growth velocity of some fetal parameters. II. Body weight, body length and head circumference. In this study, fetal growth velocities of the body weight, crown-heel length, crown-rump length and head circumference were established from 478 normal fetuses and neonates, aged 8-41 gestational weeks. The growth rates were computed by time intervals, and the velocity curves were plotted with their 95% confidence intervals. The body weight displayed an accelerating pattern of growth until 34-35 weeks and a breakdown of the growth rate afterwards, as we already observed for the brain weight in another study published in Biology of the Neonate. Decreasing patterns of growth velocity throughout pregnancy were observed for the crown-heel and the crown-rump lengths, with a marked fall at 35 weeks. The growth velocity curve of the head circumference is also decreasing, but in a more irregular way. A curious revival of growth velocity was observed in most parameters at about 38 weeks. These changes in the growth rhythm were similar to those found previously with ultrasound data.


Effects of injectable beta-carotene and vitamin A on lymphocyte proliferation and polymorphonuclear neutrophil function in piglets. Newborn piglets were injected with either (1) vehicle; (2) 20 mg beta-carotene; (3) 40 mg beta-carotene; (4) 25,000 IU vitamin A palmitate, or (5) 50,000 IU vitamin A palmitate. Blood was collected at 0.5, 1, 3, and 6 weeks of age and lymphocyte proliferation, polymorphonuclear neutrophil (PMN) phagocytosis and intracellular killing of live Escherichia coli were measured. Lymphocyte proliferation in all piglets was high at birth but decreased thereafter. Piglets injected with either high beta-carotene or vitamin A palmitate showed higher phytohemagglutinin- and pokeweed mitogen-induced lymphocyte proliferation at week 3 compared to controls. Similar results were seen with 20 micrograms/ml concanavalin A in piglets injected with high vitamin A palmitate. Phagocytosis by PMN isolated from all piglets decreased to a nadir at week 3 but returned to values observed in the newborn by week 6. Treatment had no effect on phagocytosis. Piglets injected with beta-carotene or vitamin A palmitate had lower intracellular killing ability at week 3 compared to controls. Therefore, beta-carotene and vitamin A palmitate enhanced mitogen-induced lymphocyte proliferation but reduced killing ability by PMN in piglets.


Effects of interleukin-3 on murine fetal liver hemopoiesis in utero. The normal regulation of primitive hemopoietic stem cells (PHSCS) throughout development involves locally produced factors and humoral factors. While considerable information is available on the effects of candidate hemopoietic growth factors (HGFs) on postnatal hemopoietic tissues, little is known about the regulatory events of developing hemopoietic stem cells within fetal microenvironments. Fetal hemopoiesis represents expanding populations and may be under different regulatory control mechanisms. The microinjection of purified interleukin-3 (IL-3), a candidate HGF, into 13-day-old mouse fetuses via the yolk sac, allowed us to evaluate its effects on morphogenetic events and, more specifically, on fetal liver populations using quantitative in vitro clonal assays for hemopoietic precursors. In view of the sensitivity of fetal development during the early organogenetic period, considerable care was taken to identify the stress effects of the surgical laparotomy and the microinjection procedure. Control studies, required to distinguish stress effects of surgical laparotomy and microinjection, clearly revealed that the fetal liver is a sensitive organ responding with limited tissue disorganization, reduced cellularity and erythropoietic activity, as monitored 24 h after experimental intervention. The microinjection of 15 units of IL-3 promoted a significant expansion of depleted liver hemopoietic-cell populations and had stimulatory effects on the distribution of connective tissue mast cells and absolute cell numbers, including hemopoietic precursors (erythroid, granulocyte, macrophage, megakaryocyte), compared to controls. These studies suggest that (1) fetal liver hemopoiesis is selectively sensitive to maternal stress but has an effective regenerative capacity to maintain essential hemopoiesis in utero, and (2) fetal hemopoietic cells require an ability to respond to IL-3 early in fetal development.


Maturation of jejunoileal gradients in rat intestine: the role of intraluminal nutrients. Jejunoileal gradients of intestinal function are thought to be established during the third week of life in the rat when postnatal intestinal maturation occurs. In order to investigate the normal development of jejunoileal gradients and whether either the absence of intraluminal nutrients or the form in which they are provided affected the development of jejunoileal gradients, gradients for mucosal DNA, protein, lactase and sucrase were studied in suckling rats undergoing normal weaning and compared to gradients in rats receiving no intraluminal nutrients or rats receiving nutrients in elemental form. In suckling animals, preexisting jejunoileal gradients for DNA and protein persisted through the weaning period, gradients for lactase formed by rapid decline of ileal function and sucrase gradients formed by rapid increase in jejunal activities. Intraluminal nutrients in elemental form resulted in the formation of jejunoileal gradients similar to those in intestines of normally weaned rats. The lack of intraluminal nutrients resulted in no qualitative differences in the expression of jejunoileal gradients for sucrase, but provision of elemental nutrients resulted in increased jejunoileal differences for this enzyme. The lack of intraluminal nutrients resulted in no gradients for DNA, less pronounced jejunoileal differences for protein and delayed maturational decline of ileal lactase which prevented development of jejunoileal gradients for the enzyme. These studies indicate that the formation of jejunoileal gradients in the maturing rat intestine for the parameters investigated require intraluminal nutrients regardless of the form in which they are provided for their normal expression.


A dose-ranging study of the behavioral and cardiovascular effects of CCK-tetrapeptide in panic disorder. Recent animal studies have shown that pretreatment with centrally active cholecystokinin (CCK) antagonists blocks the anxiogenic effects of CCK-tetrapeptide (CCK-4). In order to determine whether pretreatment with these antagonists can block the anxiogenic effects of CCK-4 in patients with panic disorder, a suitable challenge dose of CCK-4 must be selected. Thus, we conducted a dose range study in which patients with panic disorder (n = 29) were challenged with CCK-4 (10, 15, 20, or 25 micrograms) or placebo on two separate occasions, in a balanced incomplete block design. Patients received in random order 10 micrograms (n = 12), 15 micrograms (n = 11), 20 micrograms (n = 12), or 25 micrograms (n = 12) of CCK-4 or placebo (n = 11). CCK-4 induced anxiety and panic responses in a dose-dependent fashion. The incidence of panic attacks following the CCK-4 challenge was 17% (10 micrograms), 64% (15 micrograms), 75% (20 micrograms), and 75% (25 micrograms). None of the patients panicked with placebo. Moreover, a strong linear relationship between CCK-4 and increases in heart rate and diastolic blood pressure was found. The findings of this study suggest that a dose of 20 micrograms of CCK-4 (ED75) might be suitable for efficacy studies of CCKB antagonists and other potential antipanic drugs in patients with panic disorder.


MRS detection of whole brain lactate rise during 1 M sodium lactate infusion in rats. Proton magnetic resonance spectroscopy (1H MRS) performed in vivo on nine Sprague Dawley rats detected a threefold increase in whole brain lactate during intravenous 1 mol/L sodium lactate infusion. Significant increases in whole brain lactate were detected within 5 min after starting lactate infusion, progressively rose to a maximum level estimated at 3.2 +/- 1.5 mmol/L (all values +/- SD) immediately postinfusion, then decreased towards baseline levels during the next hr. Venous lactate concentration, increasing from 2.3 +/- 2.4 mmol/L to 43.0 +/- 8.0 mmol/L during the infusion, exhibited a steeper rise and then decreased more rapidly in comparison to changes in whole brain lactate. These data suggest MRS can be used in vivo to study acute changes in brain lactate associated with increasing blood lactate concentrations.


Lithium effect on smooth pursuit eye movements of healthy volunteers. Smooth pursuit eye movement (SPEM) dysfunctions in major affective disorder patients have been reported to be associated with lithium treatment. We report that SPEM of 13 healthy volunteers, either taking lithium (n = 7) or placebo (n = 6), were not significantly impaired by lithium. This could point to a pathophysiologic difference between affective disorder patients and a normal population.


Startle gating deficits occur across prepulse intensities in schizophrenic patients. The effects of prepulse stimuli of different intensities in inhibiting the startle reflex was assessed in 14 age-matched and gender-matched schizophrenic patients and 14 normal controls. The subjects were presented with startling stimuli consisting of bursts of white noise (106 dBA) with or without prepulse stimuli. Four intensities of prepulse stimuli were utilized: 75, 80, 85, and 90 dBA. Throughout the testing, the background noise was maintained at 70 dBA. The prepulse stimuli more effectively inhibited the startle reflex in the control group compared to the schizophrenic patients who showed deficient prepulse inhibition (gating) of the startle reflex. These results suggest that schizophrenics have impaired central inhibitory mechanisms over a fairly broad range of background noise to prepulse ratios. Further studies are needed to clarify exactly which ratios are optimal in eliciting prepulse inhibition (PPI) and in differentiating between schizophrenic and control groups.


The human brain resonance of choline-containing compounds is similar in patients receiving lithium treatment and controls: an in vivo proton magnetic resonance spectroscopy study. Lithium specifically and potentially inhibits membrane transport of choline. However, the effect of lithium on human neuronal choline content is unknown. This study was performed to determine if lithium alters the human brain choline concentration in vivo. In vivo proton magnetic resonance spectroscopy was used to compare the relative brain concentration of choline-containing compounds in seven lithium-treated patients and six lithium-free controls. No significant difference was observed in the mean relative choline resonance between the patient and control groups. Lithium treatment did not appear to alter the overall brain content of choline-containing compounds. It remains possible that a component of these compounds, particularly free choline, is elevated during lithium treatment.


Dopaminergic function in panic disorder: comparison with major and minor depression. Several lines of evidence suggest that dopamine might be involved in anxiety states. In this study, we assessed the growth hormone (GH) response to apomorphine (a dopaminergic agonist) 0.5 mg SC in nine drug-free inpatients meeting Research Diagnostic Criteria (RDC) for panic disorder who were age-matched and gender-matched with nine major depressive, and nine minor depressive inpatients. The three groups differed significantly in their mean GH peak response: 5.29 +/- 2.75 ng/ml in major depressives, 26.27 +/- 12.71 ng/ml in minor depressives, and 37.28 +/- 10.58 ng/ml in panics, with a significantly higher response in panic than in either minor or major depressive patients. These results support dopaminergic overactivity in panic disorder as compared with major and minor depression.


Familial subtypes of unipolar depression: a prospective study of familial pure depressive disease compared to depression spectrum disease. In a large multicenter effort, major depressives were systematically studied at index admission and prospectively followed up for 5 years. Primary unipolar depressives with a family history of alcoholism (depression spectrum disease) differ from depressives with a family history of depression only (familial pure depressive disease) in having more familial anxiety and somatization disorder, more divorce, more suicide attempts, more negative life events, and needed more time to recover from the index episode. In the 5-year follow-up they are more likely to develop alcoholism and drug abuse. Depressive spectrum disease patients are more likely to meet systematic criteria for neurotic depression. The data suggest that major depression is a syndrome that is heterogeneous, and may be a final common pathway of more than one familial illnesses.


Daily sleep reports and circadian rest-activity cycles of elderly community residents with insomnia. Sleep patterns were investigated in 29 insomniacs and 22 controls selected from a large sample of elderly residents of an urban community. According to sleep logs kept for a mean of 14.7 days, insomniacs took longer to fall asleep and stayed awake longer when they woke at night. In 14 insomniacs and 8 controls, wrist movements were monitored with a portable instrument. A circadian rhythm of motor activity was found in both groups, with some notable differences. Insomniacs were more active during periods of bedrest, and their mean daily rest-activity pattern--representing mean activity at equivalent times of day--was flatter. The mean level of activity over the entire experiment was similar in both groups. Insomnia in many elderly persons is thus associated with diffusion of activity over the 24-hr day, leaving bedrest periods only partially filled by rest or sleep.


EEG coherence in unmedicated schizophrenic patients: topographical study of predominantly never medicated cases. Electroencephalographic (EEG) power and coherence were compared in 11 unmedicated schizophrenics (including 9 never mediated patients) and in 15 normal controls. There was no significant difference in power between the two groups. However, interhemispheric coherence between O1-O2 was higher in the schizophrenics in the delta and beta bands, and interhemispheric coherence between T5-T6 was higher in the delta band. These results suggest that coherence is more sensitive than power for comparison of these two groups, and that cerebral function is less lateralized in schizophrenics.


Nialamide, an MAO inhibitor, increases urinary excretion of endogenously produced bufotenin in man. Nialamide, an MAO inhibitor, was given per os (PO) to a normal man who volunteered in two separate trials (total intake 300 mg and 1000 mg, respectively), and his bufotenin excretion was followed by consecutive urine samples. In both experiments the excretion rose well above the values measured from the same test subject when not taking nialamide (median 0.089 nmol/mmol creatinine, range 0.002-1.78). At its highest, the excretion was 16.5 nmol/mmol creatinine, and the maximum urinary output was 495 nmoles (56 micrograms) in 24 hr. The levels of bufotenin in plasma required for the excretion of the latter amounts are not far from those that produce psychic symptoms in man.


Plasma prolactin as a predictor of relapse in drug-free schizophrenic outpatients. A low plasma prolactin concentration has been reported to be associated with an increased risk of subsequent relapse in patients with schizophrenia. Prolactin concentration was measured in samples from stable schizophrenic men who were outpatients just prior to neuroleptic withdrawal. No relationship between prolactin concentration and time to subsequent relapse was found. Prolactin concentration may predict time to relapse only in populations characterized by specific demographic features or medication history.


Effect of m-chlorophenylpiperazine on plasma homovanillic acid concentrations in healthy subjects. In view of the abundant anatomical and functional interactions between serotonin and dopamine systems, this study examined the effect of the serotonin agonist, m-chlorophenylpiperazine (mCPP) on plasma concentrations of the dopamine metabolite, homovanillic acid. Plasma prolactin levels, body temperature, and mCPP blood level were also measured. mCPP (0.35 mg/kg) and placebo were administered orally to 10 healthy men in a randomized double-blind design. Variables were measured for 210 min after administration of capsules. mCPP raised prolactin and temperature as compared to placebo, but did not affect plasma homovanillic acid concentrations. Results suggest that mCPP does not alter dopamine function.


Good sleep, bad sleep: a meta-analysis of polysomnographic measures in insomnia, depression, and narcolepsy. Primary insomnia, major depression, and narcolepsy are usually considered to be separate disorders, distinguished by different polysomnographic profiles. But do polysomnographic data provide adequate evidence to segregate the three disorders, or might they display fundamentally the same sleep disturbance, differing only in degree? To test the viability of these two alternate hypotheses, the authors performed a meta-analysis of controlled polysomnographic studies of these disorders. A summary measure of degree of sleep disturbance was constructed from five variables: wakefulness after sleep onset, percentage of stage 1 sleep, percentage of stage 3 + 4 sleep, rapid eye movement (REM) latency, and REM density. The results of available studies for each variable were combined using a weighted average of effect sizes. An overall "sleep disturbance index" was then calculated by combining the estimates for the five above listed variables. On both the individual measures and especially on the summary index, insomnia, depression, and narcolepsy were arrayed on a simple continuum of progressively more severe sleep disturbance--congruent with the clinical observation that these disorders display progressively more disturbed sleep. These findings suggest that sleep can be disturbed in only a limited number of ways: in evaluating sleep architecture, it may not be possible to elaborate much beyond a single axis of good-to-bad sleep. Thus, polysomnographic measures may not provide adequate evidence to classify insomnia, depression, and narcolepsy as separate entities.


Mescaline-induced psychopathological, neuropsychological, and neurometabolic effects in normal subjects: experimental psychosis as a tool for psychiatric research. The psychological, neuropsychological, and neurometabolic effects of the hallucinogenic agent mescaline were investigated in 12 normal men who were volunteers. Mescaline produced an acute psychotic state 3 1/2-4 hr after drug intake, as measured by the Brief Psychiatric Rating Scale (BPRS) and Paranoid Depression Scale (PDS). The Assessment of Altered States of Consciousness (APZ) questionnaire revealed specific effects of mescaline in the visual system. Neuropsychological effects were studied with a face/nonface decision task with known right-hemisphere advantage, in which mescaline induced a decrease of functioning of the right hemisphere. In functional brain imaging using single photon emission computed tomography (SPECT), mescaline produced a "hyperfrontal" pattern with an emphasis on the right hemisphere, which was correlated with mescaline-induced psychotic psychopathology. Our findings question the validity of the concept of hypofrontality as an explanation for schizophrenic symptomatology. The study of psychoactive substances under controlled laboratory conditions has the methodological advantage of intraindividual control, and hence, minimal variability of data.


Long-term test-retest reliability of event-related potentials in normals and alcoholics. The long-term test-retest reliability of event-related potentials (ERP) measures was examined in a group of 44 controls and 71 chronic alcoholics, retested after an average of 14 months. Correlational analyses revealed moderately significant test-retest correlations for visual and auditory target N1, N2, and P3 amplitudes, with significant correlations for N1, N2 and P3 latencies. Controls and alcoholics produced similar test-retest correlations for visual and auditory ERP measures. Men and women produced equally stable ERP measures over time. Overall N1 and P3 amplitudes were most reliable in both groups followed by N2 amplitude, N1 and N2 latency, and P3 latency. The stability of ERP measures found in this study over a 14-month period in both normals and chronic alcoholics supports the use of ERPs in the study of normal and disordered cognitive functioning.


Respiratory and cardiac rhythms as windows to central and autonomic biobehavioral regulation: selection of window frames, keeping the panes clean and viewing the neural topography. Respiratory and cardiovascular processes figure importantly in biobehavioral regulation. Various cardiac and respiratory measures may, furthermore, index the activity of relatively distinct central and autonomic mechanisms. In this regard, I consider ventilatory indices of central drive and timing of respiration, and rhythmic cardiac-interval fluctuations as reflections of parasympathetic and sympathetic cardiac influences. Particular emphasis is placed upon the phenomenon of respiratory sinus arrhythmia (RSA). A number of conceptual and methodological questions are addressed concerning quantification and inference. Among others, these include the following issues: (1) What is the evidence that the three cardiac periodicities slower than heart rate reflect distinct autonomic mechanisms? (2) Does RSA reflect tonic or phasic parasympathetic regulation of heart rate? (3) Do specific quantification procedures for measuring these rhythms provide superior estimates? (4) What are some potential pitfalls for quantification and inference.


Applied use of cardiac and respiration measures: practical considerations and precautions. Cardiac and respiratory measures can be successfully applied to "real world" environments and these measures have certain advantages over both performance and subjective measures that are typically used to monitor operator state and workload. However, because of large differences between laboratory and "real world" environments one must utilize caution in directly applying laboratory data and theories to the day-to-day world environment. While most workers are highly over-trained in their jobs, laboratory subjects are often under-trained in the cognitive tasks that are used to study cognitive activity. It is possible that a substantial portion of experimental effects reported in laboratory studies is due to learning effects. In addition, relatively small changes in cardiac and respiration measures are reported to experimental manipulations in the laboratory while a much larger range of changes are reported in "real world" environments. These differences highlight questions about laboratory/real world similarities and the need to develop a database of actual work environment data. A third area of concern is the relative lack of control over the experimental situation that is the case with most applied research. The possible confounding of changes due to cognitive and physical activity levels is a major concern and strategies for overcoming these problems are suggested. The potential for valuable contributions by cardiac and respiratory measures to applied research make overcoming these difficulties worthwhile.


Respiration in psychophysiology: methods and applications. With the use of advanced equipment, respiratory measures can unobtrusively and reliably be assessed in a variety of psychophysiological research settings. New computerized analysis techniques can break down respiration into a number of components that provide valid estimates of variations in respiratory control mechanisms in the brain stem. Thus analysed, respiratory responses may vary in at least two dimensions: (A) with regard to drive and timing aspects, and (B) with regard to the metabolic appropriateness of the respiratory response. Assessment of respiratory responses may be relevant for a broad variety of research areas, including studies of the physiological effects of mental load and stress, investigations of physiological correlates of emotions and affect, and research linking physiological responses to subjective distress and psychosomatic disorders.


Measurement and analysis methods of heart rate and respiration for use in applied environments. Cardiovascular measures are used in applied settings to assess mental load. It is neither desirable nor possible to adapt the working situation to the needs of the experimenter, as can be done in the laboratory; the purpose of this paper is to discuss how invested effort, mental efficiency, and changes in cardiovascular state can be measured in applied settings, including non-stationary ones. This paper discusses the theoretical background of fluctuations in heart rate and respiration and the application of existing methods in laboratory and normal working situations. Data acquisition and analysis methods are then presented, particularly the problems of artifact detection and correction and variability indices in spectral bands in relation to the reliability of these measures. In the last section the interpretation of data acquired in environments and the specific problems inherent in such situations are discussed.


Spectral analysis of heart rate and psychological state: a review of its validity as a workload index. This paper provides some background information on concepts of mental workload and stress and the possibility of indexing them by means of spectral analysis of heart rate. A literature review of practical implementations of the technique in laboratory and field studies is provided with the purpose of evaluating its utility as a tool in the study of mental workload and stress.


Assessing pilot workload. Why measure heart rate, HRV and respiration? The application of heart rate and respiratory measures to the human factors of flight is discussed. The concept of pilot workload is related to the concept of arousal and distinguished from physical workload. Finds from studies of pilot workload using heart rate measures are reviewed for flight, simulated flight and related real-life challenges. Measurement techniques and transducers are discussed from the perspective of field measurement. Recommended procedures are presented as are directions for future work.


Research methods for measurement of heart rate and respiration. Strategies for heart rate and respiration quantification are dependent upon a knowledge of physiology and statistics. To develop appropriate and sensitive psychophysiological measures, it is necessary to understand the neural control of the autonomic nervous system and the statistical characteristics of physiological data. Moreover, it is proposed that inferences derived from physiological measures are dependent upon a statistical interaction between the characteristics of the physiological data and the measurement techniques. The steps required to analyze heart rate and respiration data are presented, ranging from data acquisition and editing to data analysis and physiological inference. A variety of techniques are described and contrasted with recommendations for future research.


[Immunological changes in chronic osteomyelitis]. We have studied several aspects of cellular and humoral immunity in 19 patients with chronic osteomyelitis (CO) compared with 11 healthy controls of similar characteristics. Patients with CO showed significantly higher values of GSR, reactive protein C (RPC), IgG and lymphocytes CD3+ and lower values of the CD4+/CD3+ ratio, as well as an hypoergic response to 7 antigens in the different cutaneous hypersensibility tests, compared with healthy controls. The rate of "in vitro" blastic stimulation by different lectins was significantly lower in the group of patients, compared with controls. These changes in the cellular immunity are not correlated with the extent, chronicity and prognosis of the disease, although we did not performed sequential studies of the immunitary condition. None of these immunological markers seem to be a better predictor of the bone infectious activity than the traditional GSR or RPC.


[Azidothymidine in the treatment of patients with human immunodeficiency virus infection and persistent generalized adenopathies]. The effectiveness and security of azidothymidine (AZT) in the treatment of patients with infection by the human immunodeficiency virus (HIV) and persistent generalized adenopathies (PGA), were assessed. Thirty six patients with HIV infection and PGA participate in the study. Eighteen were treated with AZT and the other 18 were included in the control group, since they did not accept the treatment. Both groups were homogeneous with respect to their clinical, immunological and virological characteristics. A common study protocol was used and the clinical, immunological and virological effectiveness was assessed. Lymphocyte subpopulations were quantified by flow cytometry, viral antigens were determined by sandwich-type ELISA and antibodies against viral proteins (anti-gp120, anti-gp160, anti-gp41, anti-gp24 and anti-p18) were detected by Western blot. Naranjo and Busto's algorithm was used for the causality of adverse effects. We did not observe any significant differences regarding the presence of infection and the evolution of AIDS in both groups. A positive response to thrombocytopenia was observed, more evident in patients under low doses of AZT. The small initial transitory improvement of the immunological parameters was not statistically significant. The viral antigen was not modified by the treatment. With respect to the behaviour of the several antibodies studied, no differences were observed. The initial doses of AZT had to be modified in 44% of patients due to their hematological toxicity, more frequent in the first stages of the treatment. In two patients, the treatment had to be finally discontinued due to severe neutropenia. 25% of patients showed mild to moderate gastrointestinal manifestations.(ABSTRACT TRUNCATED AT 250 WORDS)


[Immunoglobulins, beta-2 microglobulin and lymphocyte subpopulations in patients addicted to parenteral drugs (IVDA)]. Disorders in serum immunoglobulins, beta-2 microglobulin and lymphocyte subpopulations in parenterally drug-addict patients (PDAP) are analyzed. The study is divided in three parts. In the first one, a group of 33 HIV-negative PDAP without intercurrent diseases is compared with a control group of healthy non-addict persons. In the second part, the differences between a group of 58 HIV-negative PDAP and a group of 95 HIV-positive PDAP are studied. In the third part, the differences between the group of HIV-negative PDAP without associated procedures and a subgroup of HIV-positive patients including 31 asymptomatic carriers in phase II, are studied. No statistically significant differences were detected between the control group and the HIV-negative PDAP group. The HIV-positive PDAP group showed lymphocytes CD3, CD4 and a CD4/CD3 ratio statistically lower than the HIV-negative group, as well as a significant increase of the immunoglobulin IgG and beta-2 microglobulin. The same results were obtained when the subgroup of HIV-positive patients in phase II was compared with the group of HIV-negative PDAP without intercurrent diseases. According to these results, we conclude that the immunological disorders detected in PDAP patients seem to be more related with the infection by the human immunodeficiency virus and with other associated infections than with the drug-addiction itself.


[Solitary metastases of unknown origin. Apropos 64 cases]. Metastatic cancer of unknown origin is frequent subject of medical articles because of it's difficult approach. Most commonly it's present in multiple organs (more than 50%). We reported a study about solitary metastases of cancer of unknown origin and we tried to identify where are the original neoplasms that most frequently appeared in this form.


[The assessment of liver microsomal oxidative capacity via caffeine and antipyrine elimination in patients with chronic active hepatitis: preliminary results]. The hepatic metabolism of drugs in patients with chronic active hepatitis has been studied in a few occasions, finding discordant results. In our preliminary study we pretend to appraise the microsomal oxidative hepatic metabolism in 10 chronic active hepatitis HBs Ag(+) outpatients without clinical symptoms. The concentrations of both substances were measured in saliva and the caffeine and antipyrine tests of these patients were compared to control group. The difference found in the caffeine and antipyrine elimination, between the chronic active hepatitis group and the control group, have been low, without statistics significance (p > 0.1). Although, the results are preliminaries and is necessary to follow this study in many patients, probably the caffeine and antipyrine elimination are not decreased in the chronic active hepatitis without clinical symptoms. In these patients, the microsomal oxidative hepatic metabolism is not altered.


[Acute pancreatitis in thrombotic thrombocytopenic purpura. Apropos 2 cases]. Thrombotic thrombocytopenic purpura (TTP) is a syndrome characterized by fever, microangiopathic hemolytic anemia, thrombocytopenia, renal disease and neurologic changes. The development of little thrombi in the systemic microcirculation is thought to play an important role, however the etiology is unknown. We present two cases in which pancreatic disease was the first manifestation of TTP.


[Empyema due to Moraxella (Branhamella) catarrhalis]. We present a case of empyema by Moraxella (Branhamella) catarrhalis in a patient affected by a bronchogenic epidermoid carcinoma. We describe the basic clinical characteristics of the infection by Moraxella (Branhamella) catarrhalis and we confirm the low incidence of empyema by such germ.


[Myopathy associated with infection by the human immunodeficiency virus type-1 (HIV-1)]. We present a case of an active homosexual patient who was a paid donor of plasma and presented myopathy associated to infection by the human immunodeficiency virus type 1 (HIV-1). In the ultrastructural study of the muscular biopsy, we stress the formation of central intermyofibrillary "minocores", as well as the presence of nuclear extrusions in which the core is surrounded by mitochondrial accumulations. We review the clinical, histological and ultrastructural characteristics used to identity this pathology against the group of myopathies induced by zidovudine, as well as the several empirical therapies currently recommended.


[Fulminant meningoencephalitis due to Listeria monocytogenes]. We present a case of meningoencephalitis due to Listeria monocytogenes type IV in a patient in which the diagnosis of small-cells malignant diffuse non-Hodgkin's lymphoma had been established. He was hospitalized with vague clinical manifestations and without showing any neurological focus. Hours later, he presented clinical brain stem semiology. Within a 24-hour period, Listeria monocytogenes was isolated from blood and cephalorhachidian fluid (being the first time that this microorganism had been isolated for the past 20 years in our laboratory). The patient evolved to a state of coma, with Cheyne-Stokes's respiration, abolition of pupillary reflexes, decerebration position and exitus laetalis. The affection of the central nervous system has been fully documented. However, its fulminant presentation, as in the case that we present here, is less frequent, although it has been described in the literature by other authors, like Finegold et al., with cases in which the interval between the onset of the neurological focality and death ranged from 8 to 16 hours. The antibiogram showed a wide sensibility to several groups of drugs, despite the fast evolution of the disease that prevented the modification of the initial antibiotherapy (Vancomicine and imipenem).


[The quality of life and cancer].

[The quality of life and cancer]. Quality of life has been one of the main goals of medicine during the past years. With the increase of chronic degenerative diseases and the stability of the life expectancy at birth, we are beginning to think that it is not only important the number of years that we live, but as well the number of years that we live free of discapacities. On the other hand, the increase in the survival rate due to the advance of the biotechnology, as it happens with cancer, lead us to consider several psychosocial aspects neglected a few years ago.


[Single oral dose of phosphomycin trometamol versus pipemidic acid and norfloxacin in treating uncomplicated low-level urinary tract infections]. To make an assessment of the effectiveness and safety of a single oral dose compared with the conventional way of treating low-level Urinary Infections presenting no complications. This was an investigative and prospective study, using a control, a simple blind and randomised sample. SITE. Health Centres in Oviedo and Zaragoza. We looked at 150 patients and included 106 females between the ages of 16 and 75 in the study. Their clinical symptoms were compatible with low-level urinary infections presenting no complications. 49 patients were given Trometamol Phosphomicine (TP) in a single oral dose; 36 took Pipemidic Acid (PA) and 21 Norfloxacine (NFX), both of these in a dose of 400 mg twice a day for 5 to 7 days. Clinical and bacteriological assessments of the results were made 3, 7, and 28 days after treatment. A total of 75 gems were isolated: E. coli was the most common (68%). In all three groups, the main symptoms rapidly disappeared after the start of the treatment. Having considered the bacteriological and clinical findings, as well as tolerance levels, we are able to conclude that a single oral dose of Trometamol Phosphomicine is a good alternative to conventional therapy for the treatment of low-level Urinary Infections Which present no complications.


[Obesity in the province of León. Differing criteria, different prevalences]. To assess the prevalence of obesity among the adult population of our province. Cross-sectional. SITE. León Province. This was a random sample with 572 individuals over 17. They were picked from the electoral register. The confidence level was 95%, with a 3.5% length of interval. They were selected by means of proportional, multi-stage stratified sampling. The Hansen method was used for a subsequent survey of the cases missed. Socio-demographic data were collected, and weight and size measured, using standard procedures. Those who had a Quetelet index > or = 30 kg/m2 or with a relative weight > or = 120%, in line with different weight tables, were considered to be obese. Under the first criterion there was a 23.25% prevalence of obesity (CI: 19.79-26.71). The second showed long variations, running from 59.92% (CI: 55.64-64.20) to 16.17% (CI: 13.06-19.28). The prevalence of obesity among adults in our province is above that in other zones of Spain and other countries. The calculation of relative weight is not possible in an important number of cases because age or size are not properly included in the reference table. The criteria for defining someone as obese need to be unified in order to be able to make comparisons between different surveys.


[Origin and distribution of seasonal allergies. A primary care view]. To describe the seasonal allergy distribution and its relation with grass pollen in environment. Ecological and descriptive study. SITE. Primary care centre in townshipe Coslada (Madrid). PATIENTS OR OTHER PARTICIPANT: People who are attended because of seasonal allergy from 20th April to 14th June in 1990 and 1991, of the total people covered by a primary care team. No differences were found in relation to sex. 5-19 age group cumulated 66.1 and 61.2% of the episodes in 1990 and 1991 respectively. 36.2 and 31% (1990-1991) described respiratory difficulties and 21.5 and 18.8% respiratory wheezings. 82.7 and 88.4% of patients asserted that the symptoms repeated annually. Patients who related known allergy, 91.8 and 98.3% attributed their problems to grass pollen. Correlations of r = 0.5207 (p < 0.001) in 1990 and r = 0.4978 (p < 0.01) in 1991 between grass pollen the day before and the notification date were found. We found the highest care assistance in the infant-juvenile age group, the same as an important number of cases with asthmatic symptomatology. We seen a moderate relationship between the episodes notification date and grass pollen in the environment.


[Advanced booking as strategy for diminishing appointments on demand: an approach]. We looked at a group of patients included in the arterial hypertension (AHT) sub-programme, in order to analyse whether an appointment booked in advance can reduce the number of on-demand consultations requested by the patients themselves. We also assessed whether these patients sought more on-demand consultations than consumers not included in any sub-programme. A retrospective, observational, crossover study, with no random distribution. SITE. Barrio del Carmen Health Centre in Murcia. 541 consumers not included in any sub-programme and 67 patients involved in the AHT sub-programme. Patients involved in the AHT sub-programme had 9.9 +/- 8.7 on-demand consultations per year, one year before their involvement. In 1987, they had 8.3 +/- 6.2; and in 1989, 7.3 +/- 6.3 (p > 0.05). These patients did not have any more on-demand consultations than those not involved in any sub-programme, except in the 50 to 65 age-group (p < 0.001). Our model does not show that a booked medical appointment is an adequate tool for reducing demand, even when the number of booked consultations significantly increases.


[Prevalence of serological markers of the hepatitis B virus in the out-patient population of the city of Vigo]. To discover the prevalence of serological markers of the Hepatitis B virus in the out-patient population of the city of Vigo. A year-long retrospective study (from January to December, 1990) of a total of 4,581 serums belonging to patients. These came to our laboratory from patients who attended the out-patient clinics of the Hospital Xeral in Vigo. The HBs Ag, anti-HBc and anti-HBc IgM (this last if the total anti-HBc showed positive) were then determined. SITE. Primary Care and Digestive Clinics of the Hospital Xeral of Vigo's Health Area. PATIENTS OUR OTHERS PARTICIPANTS: All pregnant women were excluded from the study. The rest were men and women between 18 and 69 who provided a total sample of 4,581 serums. These findings were as follows: 954 serums had some positive marker, which presumes a prevalence of 20.8% with a Confidence Interval of +/- 0.79 (p < 0.05). Pattern 1 had 39 cases, which was 0.8% of the total number of serums studied. Pattern 2 had 141 cases (3.1%). Pattern 3 had 5 cases (0.1%). Pattern 4 had 769 cases (16.8%). Pattern 4 was the most common (80.6%), followed by Pattern 2 (14.8%) and Pattern 1 (4.1%). The rarest was Pattern 3 with 0.5%. The percentage of positive markers was greater in men than in women: this was a statistically significant difference. Overall prevalence was 20.8%. In order to reduce the number of people infected by this important disease--one which carries extremely serious socio-economic repercussions--it would be in principle advisable to vaccinate all those in high-risk groups.


Analysis of antibody reactivity in the sera of 42 patients with paraproteinaemia. The clinical expression of disease in patients with conditions in which autoimmunity is thought to contribute to the pathogenesis of disease is the result of an unfortunate combination of predisposing and environmental factors. The presence of autoantibodies showing a variety of antigen specificities in sera from many of these patients has been closely correlated with particular spectra of organ involvement or tissue destruction. Their precise role in the disease process is as yet unclear. Sera from patients with paraproteinaemia also often contain autoantibodies to a variety of cell components, although symptoms of autoimmune disease are rarely found in this group of individuals. In this study of 42 sera from patients with paraproteinaemia we have confirmed the presence of autoantibodies in 33% (13/42) of samples. Amongst the autoantibodies detected were those to human neutrophils (3), U1RNP (8) and cardiolipin (4). In five sera, the immunoglobulin class of autoantibody did not correlate with that of the monoclonal band. This study extends previous reports of the repertoire of autoantibodies present in sera from patients with paraproteinaemia.


Aberrant expression of GM1 on lymph node cells of MRL/Mp-lpr/lpr mice: influences on the autoreactivities of anti-asialo GM1 antibodies. Although changes in surface carbohydrate expression of abnormally expanded MRL/Mp-lpr/lpr (MRL/lpr) lymph node (LN) cells have previously been described, the composition and function of glycolipids present on these cells as well as the spectrum of specificity of anti-carbohydrate antibodies reactive with these cells remains obscure. Analysis of antibodies to a panel of 22 carbohydrate structures using a liposome immune lysis assay (LILA) showed that, except for anti-asialo GM2 (GA2) antibodies, marked reduction of antiglycolipid antibody levels was observed in sera from 4-mo-old MRL/lpr mice compared with these from MRL/Mp(-)+/+ (MRL/+) mice. Absorption experiments revealed that both anti-asialo GM1 (GA1) and globoside antibodies had binding capacity to MRL/lpr LN cells. To elucidate the glycolipid profiles of MRL/lpr LN cells, glycolipids were extracted from LN cells of both MRL/lpr and MRL/+ mice and analysed. A 30-fold elevation of GM1 was found in MRL/lpr LN cells compared with MRL/+ LN cells. From the results of LILA using GA1/GM1 mixed liposomes, aberrantly expressed GM1 inhibited the classical complement pathway but did not interfere with the binding of anti-GA1 antibodies to liposomal GA1. These findings suggest that a drastic GM1 increase on MRL/lpr LN cells would inhibit the action of anti-GA1 antibodies and complement on the cell surface. This may explain the escape of these cells from an activated self directed immune response.


Elevated gene expression of argininosuccinate synthetase in peripheral lymphocytes from systemic lupus erythematosus (SLE) patients. Argininosuccinate synthetase (ASS) is a rate-limiting enzyme of urea cycle and functions primarily in the liver, whereas ASS activity is hardly detected in normal lymphocytes. In this study, we examined the level of ASS gene expression in peripheral blood lymphocytes (PBL) from human SLE patients by amplification of reverse-transcribed mRNA using the polymerase chain reaction. We have demonstrated that (a) approximately 40% of SLE patients exhibited 2.5 to 5 times higher expression of ASS gene in PBL than those of healthy PBL and (b) the elevation of ASS gene expression of PBL significantly correlates with the active pathogenesis of SLE patients according to the criteria of Japanese Ministry of Health and Welfare (p < 0.001 by student's two-tailed t-test). Thus, it is suggested that ASS gene expression is a promising marker of hyperactivated lymphocytes uniquely generated in patients with systemic autoimmune disease.


Anti-glycoprotein IIb/IIIa autoantibodies are reversibly internalized into platelets in idiopathic (autoimmune) thrombocytopenic purpura. We used flow cytometry to investigate the binding of platelet-binding IgG (PBIgG) to unfixed platelets in idiopathic thrombocytopenic purpura (ITP), including that of anti-glycoprotein (GP) IIb/IIIa antibodies. Anti-GPIIb/IIIa antibodies were detected in 13/64 ITP patients using antigen-capture ELISA and immunoblotting. When unfixed platelets were incubated with ITP plasma, the PBIgG level was significantly higher than after incubation with normal plasma. When 1 microM ADP was added to unfixed platelets, which were incubated with ITP plasma and washed, the PBIgG level increased additively. GMP-140 is a constituent of platelet alpha-granules, and a monoclonal antibody directed against this protein showed weak binding to platelets after 1 microM ADP stimulation. The increase of PBIgG produced by ADP was significantly greater when ITP plasma positive for anti-GPIIb/IIIa antibody was used compared with that obtained using antibody-negative ITP plasma. This increase of PBIgG was markedly inhibited by the removal of extracellular calcium with EDTA or the dissociation of the GPIIb/IIIa complex by EDTA treatment at 37 degrees C. These results suggest that anti-GPIIb/IIIa autoantibodies are internalized by unfixed ITP platelets and stored somewhere other than the alpha-granules. This stored antibody pool can be reversibly redistributed on the platelet surface by weak stimulants such as ADP and a functional GPIIb/IIIa complex appears to be necessary for this to occur.


Enhanced tumor necrosis factor in anti-CD3 antibody stimulated diabetic NOD mice: modulation by PGE1 and dietary lipid. Administration of OKT3 anti-CD3 monoclonal antibody (mAb) to patients for transplant rejection, is associated with a distinct and often severe clinical syndrome related to massive cytokine release. Previous reports have similarly demonstrated increased levels of serum tumor necrosis factor alpha (TNF alpha) in normal mice following administration of 1452-C11 anti-CD3 mAb. In this study, we compared serum TNF alpha levels at baseline and after anti-CD3 stimulation among three groups of mice: normal BALB/c controls, pre-diabetic non-obese diabetic (NOD) mice, and diabetic NOD mice. Baseline serum TNF alpha levels, as measured by L929 cell bioassay, were 2xhigher in diabetic NOD and 3xhigher in pre-diabetic NOD compared with BALB/c. Ninety minutes after anti-CD3 mAb stimulation, serum from BALB/c controls and pre-diabetic NOD contained 2- to 8-fold higher levels of TNF-alpha as compared to untreated control mice. In contrast, following anti-CD3 mAb, there was a dramatic 20-fold increase in serum TNF alpha in diabetic NOD mice (levels > 5000 pg/ml). Additionally, anti-CD3 mAb increased the steady-state TNF alpha mRNA transcripts. Spleens from diabetic mice given anti-CD3 mAb had higher steady-state TNF alpha mRNA than spleen from normal mice similarly treated. The enhanced release of circulating TNF alpha after anti-CD3 mAb in diabetic NOD mice was abrogated by pre-treatment of mice with prostaglandin E1 (PGE1) 30 min prior to anti-CD3 mAb stimulation.(ABSTRACT TRUNCATED AT 250 WORDS)