[Properties of intracellular virus-specific ribonucleoproteins containing negative and positive hepatitis A virus RNA]. The characteristics of the intracellular virus-specific nucleocapsids containing either a negative or a positive RNA strand were studied. The immunosorption of nucleocapsids by the monoclonal antibodies against the three epitopes of NP protein failed to reveal any antigenic difference between the negative strand or positive strand-containing nucleocapsids. On the other hand, the sensitivity of virus-specific RNA in the nucleocapsids to digestion by the pancreatic ribonuclease proved to be lower for the positive strand-containing nucleocapsids.


Automatic measurement of hemoglobin F in blood obtained from patients with hemoglobin E/E and beta-thalassemia/hemoglobin/E. This paper presents an automated determination of hemoglobin (Hb) F in Hb E/E disease using Hi-Auto A1c. Blood specimens collected in Bangkok were frozen, and sent to Japan by air mail for the determination. The automatically determined values showed a high correlation with the values obtained by the classical alkali denaturation method. Hb E/E cases showed 4.24 +/- 1.75% of Hb F. On the other hand, Hb, Hct, MCV and MCH in the disease samples were lower than in the controls, but higher than those of beta-thalassemia/HbE disease. From the results it was concluded that Hb E/E could be differentiated from beta-thalassemia/HbE by combination of Hb F value and MCH or Hb in CBC.


Red blood cell and platelet induced differentiation of endothelial cells into capillary-like structure. We cultivated endothelial cells of human umbilical vein origin in the presence of red blood cells and platelet-rich plasma, and observed the phenomena which occurred in the petri dish under phase contrast microscopy. Many small particles were observed after an overnight incubation. We washed the dish two times, then added thrombin to the dish. The network of thread-like strands appeared within 10 to 20 minutes of the addition, and at the same time the small particles adhered on the surface of the strands, swelled and fused gradually to cover the surface of the strands completely. Within 30 to 60 minutes the network of the strands changed into a capillary-like structure. These phenomena were not observed if we omitted red blood cells or platelet-rich plasma. Studies by transmission electron microscopy revealed that the inner surface of the lumen of the structure was covered with cells. The cells isolated from the lumen by trypsin grew to confluence in the conventional culture medium, and showed vWF antigen on their surface. These observations indicated that the method described is useful for in vitro study of angiogenesis.


[Changed levels of substance P and somatostatin in HIV-positive children]. Substance P (sP) and Somatostatin (SOM), so as other neuropeptides can modulate neurologic and immunologic functions. sP has been described to enhance both in vitro and in vivo immunoglobulin synthesis. On the contrary, SOM has an inhibitory effect on the same activity. The modulating effect is more evident on IgA isotype. Hypergammaglobulinemia and in particular high levels of IgA is a common finding in pediatric AIDS and an imbalance among regulatory effects of neuropeptides might be suggested. In order to evaluate the plasma levels of sP in pediatric AIDS we studied 15 children with HIV infection (status P2), 10 seronegative children born to HIV positive mothers and 10 healthy children of the same age. All the HIV positive children had high plasma levels of IgG and IgA. The plasma level of sP was extremely higher in HIV positive children while no significant difference was found between seronegative children born to HIV positive mothers and healthy children. SOM was decreased in HIV positive children when compared to control groups but a significant difference was not reached. It might be supposed that HIV infection, through a dysregulation among neuropeptides interferes on immune functions and in particular on IgA synthesis. On the other hand it might be suggested that the imbalance between sP and SOM depends on the viral infection of immune cells since it has been demonstrated that SOM and other neuropeptide are synthesized by lymphoid tissue. Further studied relevance of neuropeptide disorders in pediatric AIDS.


Reconstruction of neural networks is organized by co-existence of various signal molecules or input activation in Helix pomatia L. The overlapping neural networks consisting of multifunctional neurons were shown to take part alternatively in regulation of various visceral functions or behaviour. The reconstruction of the neural networks was shown to be a function of input activation or transmitter cocktail used at the vicinity of the neurons. In rearrangement of neural networks acetylcholine (Ach), serotonin (5HT) and molluscan neuropeptide, FMRFamide were found to be effective. The modulation occurs at the level of ion channels and at least three different types, namely the Ca-, delayed K- and the unspecific cation channels were shown to be responsible for it. The modulatory action of low molecular weight neurotransmitters and peptides was more often directed to the same species of ion channels, but the maximum of channel activation can appear at different voltage ranges. The modulation of afferent and efferent pathways by signal molecules can assure the mosaic-like functioning of the units of neural networks.


Somatotopic representation of the head areas in the cerebral ganglion of the snail Helix pomatia. This study gives a detailed description of the central representation of the head regions in the cerebral ganglion after parallel nickel-lysine and cobaltic-lysine backfilling of different pairs of cerebral nerves. The backfilling of the cerebral nerves demonstrate different groups of labelled neurons in the cerebral ganglion as in the procerebrum, mesocerebrum, metacerebrum, as well as in the pleural, pedal and commissural lobes of the postcerebrum. Each head area except the anterior and posterior tentacle is represented in each labelled cell group. The tentacles are not represented in the mesocerebrum and the pedal lobe of the post-cerebrum. The different head areas in the procerebrum, metacerebrum and the pleural lobe are represented in fronto-caudal a somatotopic order.


Immunohistochemical and ultrastructural study of the enteric nervous system of earthworm, Lumbricus terrestris L. Light and electronmicroscopic data reveal the presence of a well developed nerve plexus in the gut of the earthworm. The plexus contains subepithelial solitary nerve cells and fibers and an extensive neuropil among the muscle cells. There are two types of nerve cells in the enteric plexus. The first type contains mainly dense-core vesicles, and exhibits glyoxylic-acid induced fluorescence. Since none of these cells showed serotonin immunoreactivity, they are probably noradrenergic or dopaminergic. The second type contains large dense granules, suggesting that these cells are peptidergic (neurosecretory). A part of these cells are substance P immunoreactive, however no NPY, CGRP, or proctolin immunopositive cells were found. Ultrastructurally seven types of nerve fibers can be distinguished in the neuropil. Their distribution shows great variability within parts of the enteric canal. The observation that only two types of nerve cells are located within the gut makes it probable that some of the axons are extrinsic. According to immunohistological studies they may come from the stomatogastric system or from the segmental nerves. This is further supported by the fact that there is a well-developed subepithelial serotoninergic plexus in the fore-gut. Two types of neuromuscular junctions can be visualized in the muscular layer. The first type, representing a phylogenetically earlier form, exhibits wide junctional gap and pre- or postjunctional membrane thickening. The second type is the close contact. There are significantly more junctions observed in the fore-gut than in other parts of the gut.


Activation of a divalent cation-conducting channel in heart ventricle muscle cells of the snail Lymnaea stagnalis by the molluscan cardioactive peptide FMRFamide. Ion channels with characteristics of Ca2+ channels have been found in isolated heart ventricle cells of the snail Lymnaea stagnalis. Although spontaneous Ca2+ or Ba2+ currents were seen only occasionally, spontaneous inward Na+ currents were readily observed in the absence of patch pipette Ca2+ between membrane potentials of -100 mV and +20 mV. These currents were blocked by 2 mM Ni2+, 2 mM Co2+ and 10 microM Ca2+. The channels usually ceased conducting within a few minutes after seal formation with the patch pipette and could not be re-activated with depolarizing voltage steps. However, at the cell's resting potential, 10(-8) to 10(-6) M of the molluscan cardioactive peptide FMRFamide or its analogue FLRFamide2+ applied to the cell membrane away from the patch pipette, induced unitary Ba2+ currents or, in the absence of Ca2+ in the patch pipette, Na+ currents. This suggests that a secondary messenger is involved in the FMRFamide-induced activation of these channels rather than a direct activation of a channel-receptor complex by the peptide.


Vital staining with 5(6)-carboxyfluorescein in the feeding system of Lymnaea and the selective photoinactivation of the filled neurones. Lymnaea feeding interneurones were stained with 5(6)-carboxyfluorescein by cutting the nerves containing their axons. They were located under low-intensity blue light and were healthy with normal pharmacology. The interneurones were selectively killed by high-intensity blue light.


[Palliative treatment with an endoluminal prosthesis for obstruction of the esophagus and the cardia]. Between December 1984 and September 1991, a total of 68 palliative esophageal intubations were performed in 57 patients. The mean age was 64 years, there were 40 males and 17 females. The more frequent causes of esophageal obstruction were malignant tumors of the esophagus (49%), lung (23%) and cardia (19%). Total dysphagia (24.5%), dysphagia to liquids (47%) and esophagotracheal aspiration (14%) were present in these patients. In 65% of patients insertion of the prosthesis was the first attempt at palliation, in the remaining patients it followed the failure of another type of palliation. The decision to palliate esophageal obstruction was established preoperatively in 86% of patients, in the remaining 14% it was decided at the time of surgery. The intubation by pulsion (Atkinson) [1] was successful in 57% of patients and by traction (Celestin) [2] in 86% of patients. The associated morbidity was 36% for the Atkinson prosthesis and 13% for the Celestin prosthesis, although the latter required a laparotomy. The in hospital mortality was 12.5%. The mean survival was 85 days. A normal or semi-solid diet was tolerated by over 60% of patients following intubation. Palliation of esophageal obstruction by intubation improves the quality of life but is associated with a high morbidity.


The effect of isoprinosine treatment on histamine release from murine peritoneal mast cells. The study was undertaken to examine the effect of isoprinosine treatment on in vitro histamine release from peritoneal mast cells. The experiments were done on mice which received isoprinosine orally, at a dose of 50 or 100 mg per kilogram of body weight per day, in divided doses, every 8 hours, for a period of 1-9 days. After isoprinosine treatment, there was a significant decrease of in vitro Con A-induced histamine release from mast cells. This inhibitory effect was observed in both tested groups. In the second experiment, with mice sensitized with egg albumin, the treatment with isoprinosine gave also a significant inhibition of anaphylactic histamine release from mast cells, as compared with that of control without isoprinosine treatment. Results of both experiments suggested that isoprinosine may have some inhibitory effect on mast cell activation.


Seleno-organic compounds induce interferon and tumor necrosis factor in human but not in rat or mouse lymphoid cells. The seleno-organic compounds are highly active in several anti-inflammatory assays performed in mice and rats. However, they differ from the classical non-steroidal anti-inflammatory drugs including indomethacin despite the fact that both types of drugs are inhibitors of prostaglandins and leukotrienes. Furthermore, ebselen and analogs are potent anti-oxidants in many animal cell cultures. The toxicity of the drugs is low because selenium in their structure is not bioavailable. We have discovered that the seleno-organic compounds induce interferon gamma (IFN-gamma), IFN-alpha, tumor necrosis factor alpha (TNF-alpha) and other cytokines in human peripheral blood leukocytes (PBL). Furthermore, the action of the drugs and PHA or Con A was synergistic. However, ebselen and analogs were found to be inactive as the cytokine inducers in cultured rat or mouse lymphoid cells. In contrast to their effects in human PBL, the drugs even inhibited the production of IFN-gamma after stimulation with PHA or Con A. The inhibition was dose dependent. We suggest that the induction of IFN by ebselen and analogs is species specific and it may depend on interaction of the drugs with a specific receptor and/or signal-transducing system present in human but not in some animal cells.


Histamine secretion from human mesenteric and adenoidal mast cells. Human mast cells were obtained from adenoids and mesentery by enzymatic dispersion of the tissues with the enzyme collagenase. The digestion of the tissues resulted in a cell suspension which contained 1-2% mast cells. 37.3% (adenoids) and 33.4% (mesentery) of total histamine initially present in the tissues was recovered in the dispersed cell suspensions. More than 90% of the cells were viable. The adenoidal mast cells could be sensitized passively in vitro with homologous reaginic serum and released histamine after challenge with specific antigen. Both populations of mast cells were sensitive to the action of anti-human IgE; the reversed anaphylaxis with anti-IgE was higher in mesenteric mast cells. Both examined mast cell populations were sensitive to the challenge with polymyxin B, concanavalin A and ionophore A23187, however, histamine release was only up to 10% and 20% for adenoidal and mesenteric cells, respectively. Only mesenteric mast cells responded to the action of compound 48/80. Histamine release, induced by polymyxin B, was rapid (maximal release within 5 min), maximal in the presence of 3 mM extracellular calcium ions (but also occurred in the absence of the cation).


Failure of myocardial inactivation: a clinical assessment in the hypertrophied heart. Abnormal intracellular calcium handling is observed in hypertrophied cardiac muscle and in end-stage heart failure muscle. This abnormal calcium handling results in prolongation of the calcium transient and in a biphasic calcium transient with prominent late component. In the present studies, the mechanical correlates of abnormal calcium handling were investigated in the hypertrophied human left ventricle by analysis of: 1) isovolumic left-ventricular relaxation kinetics after drastic left-ventricular unloading in patients with severe aortic stenosis after sequential balloon aortic valvuloplasty-arterial vasodilation; and 2) morphology of the diastolic left-ventricular pressure signal in patients with aortic stenosis and hypertrophic cardiomyopathy. Drastic left-ventricular unloading in patients with severe aortic stenosis by sequential aortic valvuloplasty-arterial vasodilation resulted in a slow and dyssynchronous left-ventricular relaxation pattern, as evident from a prolongation of the time constant of left-ventricular pressure decay from 46.6 +/- 12.5 to 73.2 +/- 23.3 ms (p < 0.01), and from the development of a convex downward negative dP/dt upstroke pattern. Abnormal diastolic left-ventricular pressure wave forms consisting of continuous left-ventricular pressure decay throughout diastole and/or a secondary pressure rise in mid-diastole were observed in patients with aortic stenosis and with hypertrophic cardiomyopathy. Postextrasystolic potentiation caused further slowing of this abnormal diastolic left-ventricular pressure decay, as evident from the decrease in phase of the first harmonic of a Fourier transform applied to the diastolic left-ventricular pressure wave. When an abnormal diastolic left-ventricular pressure wave form was observed at rest or after postextrasystolic potentiation, a simultaneously recorded left-ventricular monophasic action potential signal revealed the occurrence of delayed afterdepolarizations. The mechanical correlates of abnormal calcium handling or of inactivation failure in the hypertrophied human left ventricle consist of slow and dyssynchronous left-ventricular isovolumic relaxation after left-ventricular unloading and of diastolic left-ventricular aftercontractions, which hinder left-ventricular filling and which are accompanied by delayed afterdepolarizations.


Growth factors, growth factor response elements, and the cardiac phenotype. Fibroblast growth factors (FGF) and type beta-1 transforming growth factor (TGF beta 1) are pleiotropic regulatory peptides which are expressed in myocardium in a precise developmental and spatial program and are up-regulated, in the adult heart, by ischemia or a hemodynamic burden. The accumulation of trophic factors after aortic banding supports the hypothesis that autocrine or paracrine pathways might function to mediate, in part, the consequences of mechanical load. Our laboratory has demonstrated that cardiac muscle cells are targets for the action of peptide growth factors and, more specifically, that modulation of the cardiac phenotype by basic FGF (bFGF) and TGF beta 1 strongly resembles the induction of fetal cardiac genes--including skeletal alpha-actin (SkA), beta-myosin heavy chain, and atrial natriuretic factor--which are characteristic of pressure-overload hypertrophy. Unexpectedly, and despite effects like those of bFGF on five other cardiac genes, acidic FGF (aFGF) was found to repress, rather than stimulate, SkA transcription in neonatal cardiac muscle cells. The proximal 200 nucleotides of a heterologous SkA promoter were sufficient for basal tissue-specific transcription, for induction by bFGF, and for inhibition by aFGF. Thus, both positive and negative regulation by peptide growth factors can be localized to the proximal SkA promoter. Full promoter activity required each of three CC[A/T]6GG motifs similar to the serum response element (SRE) for activation of the c-fos proto-oncogene, as previously shown for SkA transcription in a skeletal muscle background. The most proximal SRE, SRE1, was sufficient in the absence of other SkA promoter sequences for efficient tissue-specific expression in cardiac myocytes (versus cardiac fibroblasts), and was stimulated by bFGF to the same extent as the full-length promoter and endogenous gene. Despite its ability to repress the SkA promoter, aFGF had no significant effect on SRE1. Both FGFs up-regulated the canonical fos SRE, to a comparable degree. Thus, SRE1 can discriminate between signals generated in cardiac myocytes by bFGF and aFGF. In cardiac myocyte extracts, two predominant proteins contact SRE1: serum response factor (SRF) and a second protein, F-ACT-1. Thus, serum response factor and F-ACT-1 are candidate trans-acting factors for basal transcription of the SkA gene in cardiac muscle cells and for induction of SkA by bFGF and, potentially, other trophic signals.


Thyroid neoplasms. Can we do any better with quantitative cytology? The differential diagnosis of thyroid neoplasms by routine cytology presents major difficulties. We therefore looked for measurable nuclear parameters that could be generated from Feulgen-stained smears obtained by fine needle aspiration biopsy (FNAB). These parameters would then be used to differentiate between benign and malignant lesions. Seventy-six patients whose cold thyroid nodule was surgically excised after FNAB and examined by a pathologist were used in this study: 56 benign, 18 malignant and 2 atypical adenomas. A set of 3,662 cells from the 33 benign cases was compared with the set of 1,712 cells from the 11 malignant nodules. Discrimination between the two populations was based on four nuclear features ranked according to their discriminating power. The first ranked was a textural parameter, followed by a densitometric and finally two other textural parameters. The rate of nuclei that could be regarded as benign was computed for each case using the ranked parameters. The average rate for the 33 benign cases was 85% (50.2 = 100%, set at the 95% confidence interval). In a prospective study of the 32 remaining patients, sensitivity and specificity were, respectively, 92% and 88%, which were higher than those obtained by conventional cytology.


Computer-aided S-phase fraction determination in DNA static cytometry in breast cancer. A preliminary methodologic study on cytologic material. This methodologic study was performed on a single-cell-cycle breast carcinoma to evaluate the feasibility of computer-aided S-phase fraction determination in DNA static cytometry. The investigation was performed on Feulgen-stained cytologic material in which the total optical density values of 1,000 consecutive, randomly selected nuclei were analyzed (MultiCycle software). A good correlation in the S-phase fraction value with flow cytometry was obtained when the G2/G1 ratio was fixed at 1.95, when the histogram data points were smoothed at least once and the coefficient of variation of the G2 peak was the same as that of G0-G1 or when a first-order S-phase polynomial model was used. The percentages of nuclei in G0-G1 and G2 were somewhat similar to those obtained with flow cytometry. The greatest discrepancy with flow cytometry was observed in the value of the coefficient of variation of the G0-G1 peak of the static cytometric data: it was at least twice as great. It always remained high despite the software options used. As for the influence of the sample size in the S-phase calculation, the software was also run on samples of 600 and 200 nuclei. When the G2/G1 ratio was fixed at 1.95, the data obtained from 600 nuclei did not differ from those obtained with 1,000 nuclei, whereas an analysis on 200 nuclei showed a substantial variation. The software also allowed calculation of the ratio of the G0-G1 peak of the neoplastic population against that of the diploid reference (DNA index), the value of which in flow cytometry was 1.0.(ABSTRACT TRUNCATED AT 250 WORDS)


Molecular structure and mechanisms of action of cyclic and linear ion transport antibiotics. As a direct result of the vision, determination, and magnetic personality of Yuri Ovchinnikov a collaboration between the Shemyakin Institute of Bioorganic Chemistry and the Medical Foundation of Buffalo was begun in the early 1970's. The collaboration generated valuable insight into the structural basis for the capture, transport, and release of ions by ion transport antibiotics and the basis for the ion selectivity of these compounds. The collaboration produced dozens of joint publications on the structure and function of cyclic and linear ion transport antibiotics, fostered fruitful exchange visits between scientists in the two Research Institutes and has been a major source of creativity in my scientific career and those of many of my colleagues in Buffalo. This review summarizes major accomplishments of the collaboration.


New techniques for three-dimensional data analysis in histopathology. This paper describes two three-dimensional (3D) analytical techniques based on 3D mathematical morphology that have been found useful in quantifying the 3D spatial distribution of S-phase cells in a tubular tumor of the human breast. One technique is based on determining the normalized radial distribution of the S-phase cells with respect to the central axis of the tumor. The other technique is a novel extension of the polyhedra of Voronoi to quantify the distribution. The Voronoi polyhedron of a given S-phase cell nucleus is that polyhedron of minimal volume defined by planes all of which are perpendicular bisectors of the vectors extending from the given cell to all other S-phase cells in the tumor. Methods are demonstrated for generating these polyhedra and for histogramming their volumes. An illustration is given of using the histogram to sort the S-phase cells according to their 3D positional relationships. Displays showing the sorted cells in 3D and their associated Voronoi polyhedra are provided.


Prevalence of hepatitis C virus antibody among Korean adults. To estimate the prevalence of hepatitis C virus (HCV) infection among Korean adults and to present the putative route of HCV transmission among them, serum samples from 4917 adults older than 20 years of age were tested for antibody to HCV (anti-HCV), and histories of blood transfusion and other pertinent information were obtained by self-administered questionnaires. The overall prevalence of anti-HCV was 1.7%; prevalence was 1.4% in subjects with normal levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT), 3.3% in those with slightly elevated and 5.9% in those with markedly elevated levels of the enzymes. The prevalence of anti-HCV increased with increasing age (P < 0.01), but was not associated with blood transfusion. The present study suggests that the prevalence of HCV infection was 1.4% and that the major routes of HCV transmission may be other than blood transfusion in healthy Korean adults.


Production and characterisation of a human monoclonal thyroid peroxidase autoantibody. A human-mouse hybridoma has been produced by fusion of Hashimoto thyroid lymphocytes with the mouse myeloma line X63-Ag8.653. The cloned hybridoma secreted 2.5 micrograms per 10(6) cells per day of an IgG kappa thyroid peroxidase (TPO) autoantibody (2G4) with high affinity (2.5 x 10(9) molar-1) and specificity for human TPO. 2G4 did not react with lactoperoxidase, horseradish peroxidase or human myeloperoxidase or with porcine TPO or with human thyroglobulin. Plastic tubes coated with 2G4 bound about 50% of 125I-labelled human TPO added and the binding was inhibited by IgGs prepared from 18/18 TPO autoantibody-positive sera. This indicated that all 18 sera contained autoantibodies which recognised the same (or closely related) epitope as 2G4. Plastic tubes coated with IgGs from different TPO autoantibody-positive patient sera also bound 125I-labelled TPO but inhibition by 2G4 in this system was not complete. This suggested that the sera contained at least 2 types of TPO autoantibodies, with only one type of autoantibody reactive with the same epitope as 2G4.


Lymphocytes utilise sialylated surface molecules to accumulate in developing lesions of autoimmune encephalomyelitis. The accumulation of desialylated radiolabelled normal spleen cells and non-neuroantigen specific CD4 T-lymphocytes was measured in the lumbosacral spinal cord of Lewis rats with autoimmune encephalomyelitis (EAE) induced with myelin basic protein in Freund's adjuvant. The labelled cells were preincubated with sialidase and thoroughly washed prior to intravenous injection into rats exhibiting early clinical signs of EAE. Four hours later, the rats were killed and blood and spinal cord samples were radioassayed. Compared with untreated cells, desialylation markedly reduced the accumulation of both normal spleen cells and memory T-lymphocytes in the spinal cord, despite similar levels of cells being present in the blood. In another experiment, the accumulation of desialylated, macrophage-depleted spleen lymphocytes was measured during the onset, recovery and short-term "relapse" phases of acute EAE. Again, compared with controls the accumulation of desialylated lymphocytes was always significantly less, despite similar numbers of cells in the circulation. Lastly, intravenous injections of sialidase produced delayed onset of both clinical and histological signs in rats with passively-transferred EAE. These data confirm and extend previous findings, using a different animal model, that sialyl residues on the lymphocyte surface are important to the accumulation of such cells at inflammatory sites in the central nervous system. The possible relevance of these findings to human demyelinating disease is discussed.


[A case of hepatocarcinoma preceded by several years by "isolated" increase in alphafetoprotein]. Hepatocarcinoma (HCC), the most frequent malignant hepatic neoplasia, is sometimes difficult to diagnose at an early stage since the symptoms may be attributed to concomitant hepatic cirrhosis. The assay of alpha-fetoprotein associated with an ultrasound examination of the hepatic parenchyma is an important screening tool for high-risk patients. Ultrasound examination is considered the most sensitive method and alpha-fetoprotein is a supplementary diagnostic tool. Elevated alpha-fetoprotein only occasionally precedes morphological anomalies and even in these cases the neoplastic aspect emerges within a short period of time. The case reported here illustrates the "astronomic" increase of alpha-fetoprotein in a high-risk patient for HCC (positive HBsAg cirrhosis) without the manifest appearance of any instrumental or histological data confirming the presence of the tumour for two years. When the tumour was identified in instrumental tests it had spread throughout the entire hepatic parenchyma in a form which could no longer be treated using any form of therapy. The case reported here emphasizes the diagnostic value of alphafetoprotein in high-risk patients for HCC, even in the prolonged absence of all other data regarding neoplastic transformation.


Autoantibodies to the Ro/SSA antigen are conformation dependent. I: Anti-60 kD antibodies are mainly directed to the native protein; anti-52 kD antibodies are mainly directed to the denatured protein. Recent studies have shown that Ro/SSA autoantigen is heterogeneous and the autoanti-Ro/SSA response is correspondingly heterogeneous. There are two isoform families; the 60 kD forms and the 52 kD forms. We studied the antigenic difference between the native and denatured Ro/SSA isoforms and found that the autoanti-Ro/SSA response to the native 60 kD antigen is quite homogeneous. All anti-Ro/SSA sera recognize the native kD antigen regardless of the reactivities to the 60 kD band on the Western blot. Surprisingly, no anti-Ro/SSA sera without anti-La/SSB reacts with the native 52 kD Ro/SSA, although sera with both precipitating anti-Ro/SSA and anti-La/SSB can immunoprecipitate the native 52 kD antigen. Anti-Ro/SSA sera exist which react exclusively with the native 60 kD Ro/SSA protein (10/43, 23%) while no anti-Ro/SSA sera have been found which react exclusively with the denatured 52 kD Ro/SSA antigen. In sera with anti-Ro/SSA precipitins alone, only antibody to the denatured 52 kD Ro/SSA molecule is found! In sera with anti-Ro/SSA and anti-U1 RNP precipitins, no antibody to either native or denatured 52 kD Ro/SSA is found, while in sera with both anti-Ro/SSA and anti-La/SSB precipitins, antibodies to both the native and denatured forms of 52 kD Ro/SSA are present. These data suggest that the anti-Ro/SSA response to the 60 kD molecule is driven by the native 60 kD Ro/SSA molecule while the molecular identification of the antigen drive in the anti-52 kD Ro/SSA response is unknown.


The role of intermediate filaments in early Xenopus development studied by antisense depletion of maternal mRNA. The effects of depleting a maternal cytokeratin mRNA on the developing embryo are described. Cytokeratins are members of the intermediate filament family of cytoskeletal proteins, and are expressed in a cortical network of the superficial cytoplasm of the oocyte. After fertilisation, a new cortical network is built up, which comes to occupy only the most superficial cells of the blastula. The maternal cytokeratin mRNA is abundantly translated, both during oogenesis, and during oocyte maturation and after fertilisation. Depletion of the mRNA results in depletion of the cortical filaments at the blastula stage and leads to gastrulation abnormalities. We discuss the various possible control experiments required for antisense oligo depletion studies and the implications of these results for cytokeratin function.


Effect of benzyl-alpha-GalNAc, an inhibitor of mucin glycosylation, on cancer-associated antigens in human colon cancer cells. Many cancer-associated antigens are present on mucin glycoproteins. These include peripheral antigens such as sialyl Lea and sialyl Lex and core region carbohydrate antigens such as T, Tn, and Sialyl Tn. We have recently described an inhibitor of mucin glycosylation, benzyl-alpha-GalNAc. The purpose of this study was to determine its effect on expression of mucin carbohydrate antigens. HM7 colon cancer cells were treated for 2 days in culture with 2 mM benzyl-alpha-GalNAc. This treatment did not affect viability or doubling time, but inhibited synthesis of [3H]glucosamine-labeled mucins. There was also secretion of benzyl-oligosaccharides and a decrease in the proportion of long oligosaccharides on 3H-labeled mucins. Mucins were purified from spent media by gel filtration and assayed for binding of monoclonal antibodies and lectins. Mucins from benzyl-alpha-GalNAc-treated cells had increased binding of peanut agglutinin (specific for T antigen, Gal beta 3GalNAc) and Vicia villosa agglutinin B4 (specific for Tn antigen, GalNAc alpha-Thr/Ser), but decreased binding of monoclonal antibodies 19-9, SNH3, and 91.9H (specific for sialyl Lea, sialyl Lex, and sulfomucin, respectively). Treatment of the cells with benzyl-alpha-GalNAc also decreased their binding to E-selectin (ELAM-1), which recognizes sialyl Lea and sialyl Lex. Thus, benzyl-alpha-GalNAc treatment, which decreases the level of peripheral carbohydrate carbohydrate antigens on mucins with accumulation of core region antigens, may be useful in modifying the immunological and biological properties of colon cancer cells.


Measurement of uptake and incorporation of nucleic acid precursors by preimplantation mouse embryos after development in vivo and in vitro. To assay DNA and RNA synthesis by developing mouse embryos in vitro and in vivo, we measured the uptake and incorporation of 3H-thymidine and 3H-uridine by morulae and blastocysts. We also evaluated the effect of adding EDTA to the culture medium on the uptake and incorporation of nucleic acid precursors by blastocysts. Thymidine and uridine incorporation increased after morulae developed into early blastocysts both in vitro and in vivo. However, the rates of uptake and incorporation were significantly lower by embryos grown in vitro than by those grown in vivo. The ratios of incorporation to total uptake were similar in embryos grown in vitro and in vivo. EDTA (100 microM) added to the culture medium significantly increased the incorporation of uridine into RNA by blastocysts grown in vitro (P < 0.01) but did not increase the total uptake of uridine. These observations showed that both DNA and RNA synthesis increased during the early development of preimplantation embryos and that those activities were reduced in embryos undergoing development in vitro. The results also suggested that through the mechanism of EDTA effect in embryo culture remains unknown, it appeared to reduce the retardation of RNA synthesis by embryos cultured in vitro through a selective stimulation of uridine incorporation.


[The diagnostic value of erythrocyte polyamines (EPA) in prostatic adenocarcinoma (PA): apropos of 100 patients]. Spermidine and spermine are ubiquitous polyamines which are intensely metabolised in the prostate. Polyamines are involved in the processes of proliferation and differentiation of normal and neoplastic cells. As the erythrocyte levels of these polyamines are correlated with the intratumoral levels, we assayed EPA in 45 controls, 66 patients with benign prostatic hypertrophy and 100 patients with prostatic cancer. Erythrocytic polyamines are markers of proliferation and prostatic metastases and can be used to distinguish between hormone-sensitive and hormone-resistant patients. Although non-specific, polyamines constitute circulating markers of the state of tumour proliferation of a given patient and definitely have a prognostic value which needs to be evaluated by further studies.


[Progression after radical prostatectomy of cancer of the prostate: prognostic criteria and the role of PSA in monitoring]. Progression after radical prostatectomy, evaluated by a rise in plasma PSA and/or the appearance of a pelvic nodule positive on biopsy and/or the presence of bone metastases confirmed by bone scan, was studied in a series of patients with prostatic cancer with a follow-up of between 6 months and 5 years. The progression-free survival rate was 86% at 1 year and 60% at 5 years. A progression-free survival rate and the relative risk of progression were established on the basis of the morphological characteristics (anatomical stage, tumour volume, seminal vesicle invasion, condition of the prostatic capsule and lymph nodes, positive resection margins at the apex) and histological features (Gleason's score) of the cancer, allowing determination of the influence of prognostic criteria on the outcome. The positive resection margins at the apex were due to preservation of the nervi erigentes. The preservation of the neurovascular pedicles may not be justified in the case of a tumour confined to the prostatic apex.


[Benign and malignant Brenner tumor of the ovary: a clinicopathological and immunohistochemical study]. The clinicopathological and immunohistochemical features of 7 cases of benign and malignant Brenner tumor of the ovary, including 5 benign and 2 malignant tumors are described. Microscopically, all of the benign cases were composed of both epithelial nest and fibrous stroma. Two cases of the malignant Brenner tumor showed that the histologic features resembled the structure of non-keratinized squamous carcinoma or transitional cell carcinoma. Immunohistochemistry showed that tumor cells of the epithelial nest were keratin and EMA positive in 7 cases; CEA-positive in 5 cases; and negative in 2 cases of benign Brenner tumor. The results indicated that Brenner tumor is an epithelial neoplasm in nature. The diagnostic criteria and histogenetic origin are discussed.


[Sarcomas of the uterus: immunohistochemical characterization and diagnosis]. Pathologic findings and immunohistochemical characterizations of 18 cases of uterine sarcomas were studied. In endometrial stromal sarcoma (ESS), 5 out of 10 cases had ovarian sex cord-like pattern and 4 out of 10 cases had smooth muscle differentiation. Immunohistochemical findings showed vimentin, desmin and cytokeratin positive in 9/10, 6/10, 2/10 cases respectively which reflects that ESS may differentiate into both epithelium and muscle components morphologically. In malignant mixed Mullerian tumors (MMT), its carcinomatous structure may be positive about vimentin, and its sarcomatous structure may be positive to the epithelium markers, which indicates that both the sarcoma and carcinoma structures have possibly a common origin. It is considered to be of value for the diagnosis of MMT, if the tumor has differentiated both epithelium and mesoderm components or to be positive to myoglobin, NSE* in immunoreaction, accompanying with the morphologic characterizations of the tumor.


[The inhibitory effects of catechin derivatives on the activities of human immunodeficiency virus reverse transcriptase and DNA polymerases]. Catechin derivatives including (-)-epicatechin gallate (ECG), (-)-epigallocatechin gallate (EGCG), (-)-epigallocatechin (EGC) and green tea extract (GTE) were found to inhibit the activities of cloned human immunodeficiency virus type 1 reverse transcriptase (HIV-1 RT), duck hepatitis B virus replication complexes reverse transcriptase (DHBV RCs RT), herpes simplex virus 1 DNA polymerase (HSV-1 DNAP) and cow thymus DNA polymerase alpha (CT DNAP alpha). EGCG and ECG were shown to be very potent inhibitors of HIV-1 RT. According to the IC50 values for HIV-1 RT, these compounds can be ordered as EGCG 0.0066 mumol/L > ECG 0.084 mumol/L > GTE 0.1 microgram/ml > EGC 7.2 mumol/L. DHBV RCs RT was the least sensitive to these compounds. Kinetic study showed that EGCG exerts a mixed inhibition with respect to external template inducer poly (rA).oligo (dT) 12-18 and a noncompetitive inhibition with respect to substrate dTTP for HIV-1 RT. Bovine serum albumin significantly reduced the inhibitory effects of catechin analogues and GTE on HIV-1 RT. In tissue culture GTE inhibited the cytopathic effect of coxsackie B3 virus, but did not inhibit the cytopathic effects of HSV-1, HSV-2, influenza A or influenza B viruses.


[The application of monoclonal antibodies (McAbs) against human serum IgA in single radial immunodiffusion (SRID)]. Among many McAbs to human IgA, two McAbs (A1A10 and A1C9) against two determinants were selected and mixed. They formed opaque precipitation rings with IgA in agarose containing 2% PEG 6000. SRID with our mixed McAbs or PcAbs was used to determine serum IgA concentration in 40 undiluted serum specimens: a comparison of the two showed similar results, with a correlation coefficient of 0.9493 (P < 0.001). We conclude that mixed McAbs A1A10 and A1C9 can replace PcAbs in SRID for the measurement of human total serum IgA.


Photosensitivity of 8BrcGMP-induced conductance in ROS-excised patches. Inside-out patches from ROS plasma membranes contain the basic enzymes of the phototransduction cascade. Similar to a native photoreceptor cell, such patches are capable of responding to light, the effect of which suppresses the cGMP-activated current. Photoresponses are observed only in the presence of GTP, whereas ATP essentially accelerates the current recovery to a dark level. Photoresponses are also observed in the presence of 8BrcGMP. Phosphodiesterase (PDE) hydrolyzes 8BrcGMP two orders of magnitude slower than cGMP, so the light inhibition of the 8BrcGMP-induced current cannot be accounted for by PDE activation. It seems that activity of cGMP-gated channels depends not only on cGMP concentration, but is additionally controlled by some other regulatory mechanisms.


[Hepatocellular adenoma with a high level of alpha fetoprotein. Apropos of a case observed in the Central hospital of Yaoundé, Cameroon]. The authors report a case of a young girl aged 23 years, suffering from hepato-cellular adenoma associated with a high level of alpha-foeto-protein. The patient after having been operated recovers and the level of alpha-foeto-protein returns to normal.


Optical principles of loupes.

Optical principles of loupes. The use of magnification to improve precision in dental procedures goes back more than a century. However, inadequate understanding of the optical principles employed in the various types of dental loupes can lead to poor fit and adjustment and, eventually, unnecessary eye strain. Eye movement, depth of field, working distance, illumination, crossover point and exit pupil are defined and discussed relative to choosing dental loupes.


The anterior single-tooth implant restoration. Placing an anterior single-tooth implant requires thorough planning, informed abutment selection and precise surgical technique. Planning procedures are outlined; abutments, including UCLA's non-segmented, Nobelpharma's single-tooth, CeraOne and non-rotational, implant innovations' pre-angled and non-rotational and DIA's anatomic, are described, with indications and contraindications; and placement considerations are discussed.


Precision attachment removable partial dentures. Although added preparations and skill are required to provide precision attachment removable partial dentures, more favorable esthetics and load distribution may outweigh the disadvantages. Basic design principles, categories (intracoronal, extracoronal, stud, bar and plunger) and selection of precision attachments are reviewed.


Microscopes in endodontics.

Microscopes in endodontics. The introduction of microscopes in clinical and surgical endodontics is fairly recent. Potential applications, including removal of fractured instruments, localizing calcified canals, soft tissue management and root-end procedures, are reviewed.


Rapid identification of Actinobacillus actinomycetemcomitans based on analysis of 23S ribosomal RNA. Actinobacillus actinomycetemcomitans is a key microorganism in the pathogenesis of several different forms of periodontal diseases. Identification of this bacterium from clinical specimens may often be complicated by the fact that the colony morphology on TSBV selective medium closely resembles that of Haemophilus aphrophilus and a key differentiating characteristic, catalase reaction, may be variable. Recent genetic studies have shown that the 23S ribosomal RNA molecule is split into two smaller forms in A. actinomycetemcomitans, but is intact in H. aphrophilus. Based on this finding, we describe a new, rapid method for identifying A. actinomycetemcomitans in which single colonies isolated from culture on TSBV agar in 5% CO2 in air are lysed, electrophoresed on 1.5% submarine agarose gels and visualized by staining with ethidium bromide. Using this assay, A. actinomycetemcomitans can be easily distinguished from morphologically similar colonies such as H. aphrophilus strains by differences in 23S rRNA within 2 h.


[Biological actions and therapeutic perspectives of double stranded polyribonucleotides: a reappraisal]. Double-stranded polynucleotides, which are composed of two complementary homopolyribonucleotides containing no genetic information, are synthetic molecules capable of mimicking the action of natural double-stranded RNA or viral RNA on cells. Double-stranded polyribonucleotides act as an alarm system alerting the cell to the presence of an external aggression, e.g. a viral attack. In addition, polyribonucleotides have a more active function in that they trigger cell defense processes through activation of a family of genes, of which some encode cytokines, activation of cytoplasmic enzymes involved in antiviral mechanisms or signal transduction, and activation of nonspecific immune responses. Double-stranded polyribonucleotides containing one mismatched base pair per helix have been found to be especially interesting. The best known example is poly(I).poly(C12U), also called ampligen. Poly(I).poly(C12U) is capable, in experimental models, of limiting the development of viruses (including HIV), reducing tumor growth, eliminating metastases, and, according to one report, preventing steady declines in T-cell counts in HIV-positive patients. Therapeutic doses used in the USA as an experimental drug induced little toxicity. In vitro, poly(I).poly(C12U) acts synergistically with interferon, interleukin 2 or AZT, suggesting that these latter drugs may be effective in lower, less toxic doses when used in combination with poly(I).poly(C12U). The therapeutic activity of poly(I).poly(C12U) holds promise. More extensive prospective studies of this agent are warranted.


Intraovarian regulation: the IGF-I example. A large body of information now supports the existence of an intraovarian system of insulin-like growth factors (IGFs) replete with ligands, receptors, and binding proteins. The intraovarian IGF system is most likely concerned with the amplification of gonadotrophin hormonal action, other potential regulatory roles remaining speculative at this time. There is every reason to believe that work in the next few years will yield the insight necessary to establish whether or not IGFs are truly indispensable to ovarian function; this remains to be demonstrated through selective ablation of ovarian IGF-I gene expression and the examination of the impact of such manipulation on reproductive potential.


[Effects of amiodarone on kinetics of procainamide and its major metabolite disposition in rabbit]. The effects of steady state amiodarone on the kinetics of procainamide (PA) and its major metabolite, acetylprocainamide (NAPA), disposition in rabbits were examined. When PA was given alone, the pharmacokinetic parameters alpha, beta, Kmo, K1m.V1.V-1m, were 0.20 +/- 0.04, 0.016 +/- 0.003, 0.027 +/- 0.01, and 0.043 +/- 0.02 min-1, respectively. When PA was given in combination with amiodarone, the corresponding values were 0.37 +/- 0.08, 0.0059 +/- 0.002, 0.0096 +/- 0.004, and 0.016 +/- 0.003 min-1, respectively. The results indicated that the disposition kinetics of both PA and NAPA were significantly changed by steady state amiodarone.


[Early biological markers of anaphylactoid reactions occurring during anesthesia]. Three markers of in vivo histamine release, i.e. plasma histamine and tryptase, and urinary methylhistamine, were assessed using sensitive radioimmunoassays in 18 patients who had experienced an adverse reaction to an anaesthetic agent. Controls were obtained from 35 patients following a general anaesthetic, which included a muscle relaxant, and who remained free from any adverse reaction. A first blood sample was obtained from all 18 patients a mean 25 +/- 26 min after the reaction, and a second one in thirteen a mean 120 +/- 65 min after the reaction. Ten patients had had a life-threatening reaction. Plasma histamine levels were increased in all these cases, and tryptase concentrations in 9 out of 10. Urinary methylhistamine rarely reached pathological levels (4 out of 10). Skin tests were positive in the four tested patients. Plasma histamine concentration was still high in 8 cases thirty minutes after the reaction, and remained increased for more than 2 h in two patients. Among the other eight patients with a moderate reaction, 3 had high histamine levels, with normal or weakly increased tryptase concentrations, and normal urinary methylhistamine. Two of these patients had positive skin tests. There were no abnormal findings in any of the investigations carried out in the other five patients, except for a slightly positive skin test to atracurium in one patient. Plasma histamine had a higher sensitivity than tryptase levels. Methylhistamine concentrations were only rarely of interest. There were no false positives with the three investigated markers.(ABSTRACT TRUNCATED AT 250 WORDS)


Combined bromodeoxyuridine immunohistochemistry and Masson trichrome staining: facilitated detection of cell proliferation in viable vs. infarcted myocardium. Cells in the S-phase of the cell cycle can be identified in tissue sections by immunohistochemical localization of the thymidine analogue bromodeoxyuridine (BrdU). Generally, a single counterstain is used to visualize the underlying tissue; however, interpretation of morphologic detail is often difficult. We have utilized BrdU to localize proliferating cells in myocardium exposed to angiogenic mitogens. To facilitate identification of labelled nuclei in the context of infarcted vs. viable myocardium, BrdU immunohistochemistry was followed by a modified Masson trichrome stain. The time of exposure to the counterstains and the wash protocol were re-revised, permitting clear identification of the labelled brown nuclei against a background of red viable myocardium vs. blue infarct. The combined technique also provides color contrast suitable for computer-based image analysis.


Golgi method without osmium tetroxide for the study of the central nervous system. A variant Golgi technique was developed that consisted of substituting osmium tetroxide with formaldehyde as the initial fixative in intracardiac perfusion, along with the addition of glacial acetic acid to the chromating fluid. This procedure avoids disposal of dangerous waste substances into the environment. Other advantages include 1) reduction of cost, danger to lab workers, and risk of disruption of the tissue slices during their handling by eliminating the osmium tetroxide, 2) clear tissue background, 3) greater quantity of impregnated neurons than in the classical procedure, with distinct morphological details easily identified even in gross sections and 4) reduction in processing time.


Anti-centromere antibody and CREST syndrome in patients with primary biliary cirrhosis. Anti-centromere antibodies (ACA) in 41 sera from patients with primary biliary cirrhosis (PBC) were analyzed by an immunoblotting method and the correlation between the presence of ACA and the clinical features in these PBC patients was studied. In 10 of 16 ACA-positive PBC patients, one or more clinical features of CREST syndrome (PBC-CREST) were found. Statistical differences were observed in age at disease onset, serum levels of IgM and total bilirubin and titer of anti-M2 antibody, between PBC-CREST patients and the PBC patients without CREST symptoms (PBC-non CREST). By immunoblotting analysis, three major epitopes of ACA were identified at 18 kD, 80 kD and 140 kD polypeptides. The 18 kD polypeptides were detected in all 16 ACA-positive PBC patients. From these results, it is suggested that ACA-positive PBC-CREST patients can be separated from ACA-negative PBC-CREST and PBC-non CREST patients.


Primary mediastinal diffuse large cell lymphoma initially presented with pericardial infiltration. A 33-year-old Japanese woman visited our hospital with progressive dyspnea. A chest roentgenogram and a chest computed tomogram revealed a mediastinal tumor and pericardial effusion. Ultrasound cardiography revealed the existence of cardiac tamponade. She was successfully treated with emergency pericardial drainage, thoracocentesis and chemotherapy. The histological diagnosis of diffuse large B cell lymphoma was established with the tumor specimen obtained by transcutaneous fine needle aspiration biopsy and through immunohistochemical analysis. This is a rare case of primary mediastinal diffuse large cell lymphoma with initial presentation of a symptom related to pericardial effusion.


Polysaccharide storage myopathy associated with recurrent exertional rhabdomyolysis in horses. A polysaccharide storage myopathy is described in nine Quarterhorses, Quarterhorse crossbreds, American Paints and Appaloosa horses which had a history of recurrent exertional rhabdomyolysis. Muscle biopsies were characterized by high muscle glycogen concentrations with up to 5% of type 2 muscle fibers containing inclusions which stained positively with the periodic acid Schiff (PAS) stain. The inclusions were classified as an acid mucopolysaccharide, based on their histochemical staining characteristics. Ultrastructural studies revealed that the inclusions were composed of beta glycogen particles interspersed among arrays of filamentous material. In addition, many type 2 fibers contained multiple subsarcolemmal vacuoles. These vacuoles stained lightly with eosin and did not stain positively with PAS. Centrofascicular atrophy and necrosis of scattered type 2 fibers were present in biopsies from some horses. No glyco(geno)lytic enzyme deficiencies were identified using a biochemical screening test for anaerobic glycolysis. Attempts to measure branching enzyme activities in both affected and control samples were unsuccessful, employing methods developed for human muscle. The polysaccharide accumulation in these horses may represent a hereto yet undefined metabolic disorder of skeletal muscle.


[Neurohormonal regulation of the development and regenerative processes of bone and cartilage tissues]. Neurohumoral regulation of cartilaginous and osseous tissues was investigated in different stages of ontogenesis. It has been shown that these tissues are targets practically for all hormones produced by the body. The reciprocal reaction of osseous and cartilaginous tissues to different concentrations of hormones was estimated. It has been also demonstrated that the action of hormones in these tissues manifests itself in accordance with the general regularities and that their effects are strictly specific for the test target tissues. An opinion is advanced about the breach of the endocrine situation in the body as a risk factor of the development of dystrophic processes in cartilages.


[Hereditary (dysplastic) diseases of the joints]. Using the intentional procedures to reveal systemic semantic interrelations between different phenomena that form the causative-sequential chain in joint diseases of dysplasia genesis and assuming the pathology theory laws as a basis, applying the methodologies of different interdisciplinary sciences and the technology of cognitive simulation, the conceptual model "joint dysplasia--dysplasia arthrosis" was developed. This model depicts the characteristic features of joint dysplasia. The use of the model permitted one to base and describe a group of inherited (depressional) diseases of joints as an independent scientific orthopedo-arthrological problem. The application of the conceptual model confirmed its high cognitive and reforming role. Using this model on a knee joint new syndromes of dysplasia have been described; the system of roentgenometric diagnosis of dysplasia anomalies and new methods of surgical intervention have been elaborated; the foundation has been laid of prophylactic surgery aimed at the exclusion or at the minimizing of the risk of gonarthrosis in persons suffering from different inherited syndromes.


[Stress and the body].

[Stress and the body]. The paper deals with new data on the influence of stress on the neurophysiological, hematological, cardiological, hormonal and trophic processes occurring in the body of man and animals. Experimental emotional stresses which offer an opportunity to effectively explore systemic stressful reactions are described. It is shown that neurogenic stress plays the pathogenic role in the onset of blood disorders, in activation of the adrenergic system, and in disorders of the conduction system of the heart. Emphasis is laid on the processes of dystrophy and destruction in osseous and cartilaginous elements of the bones and joints. The author base the role of emotional stress (according to P. K. Anokhin) in the genesis of diseases of motion organs (osteochondrosis, arthrosis, parodontosis).


[Pathological changes induced by environmental chemical pollutants in tissues of the locomotor system]. The most widespread contaminants that accumulate in the environment due to the activity of man are mercury, lead, phenol and sulphur compounds. They have a harmful effect on the body and to an even greater extent on its progeny. A study was made of structural and functional changes in supporting (hyaline cartilage, bone) and hemopoietic (bone marrow) tissues under conditions of lead and phenol poisoning of albino rats and guinea pigs. The methods applied were histochemistry, historadiography, and electron microscopy. It has been established that lead can be accumulated by tissues (particularly by cartilage and bone), inducing disorders of plastic and energy metabolism in the cell. This is attended by the growth delay, osteoporosis and inhibition of normal and reparative osteogenesis. The toxic effect of phenol on the cells manifests by inhibition of the enzymic activity; there is evidence of disturbance of oxidative phosphorylation and of the mechanisms of specific biosynthesis and energy metabolism in the cells. Under conditions of phenol intoxication the chondrocytes and osteoblasts show disorders in the biosynthesis of collagen and sulphated glycosaminoglycans with an evident disproportion of their macromolecules in the cartilage and bone matrix.


[New approaches to developing drugs with cartilage protective properties]. The conjuncture and technological analysis was made of the current status and approaches to further development of the drugs--chondroprotectors on the basis of glycosaminoglycans. The concept of the goal-oriented search of the new type of drugs for the treatment of diseases of joints and other pathologies was worded on the basis of the systematization and summarization of the reported data and the results of own investigations. This concept rests on the current views of biochemistry of the nature of metabolism and the mechanisms of its regulation. The approach suggested lies in obtaining specific products--protein fragments (polypeptides) that contain oxidized functional groups not found, as a rule, under natural conditions in normal physiological state. The conversion of protein fragments to a higher degree of oxidation means the conversion to the metabolic state in which the possibility of regulating disturbed metabolism is provided. In order to realize the given approach, it is proposed that the oxidation-hydrolytic modification of human placenta may be performed with the aid of chlorine dioxide or direct synthesis. The results of studying the physicochemical and pharmacological properties of the products obtained are presented. Animal experiments have demonstrated a powerful hitherto unknown pharmacotherapeutic action in a dose of 0.3 mg/kg, namely chondroprotective, resolving, reparative, antiinflammatory, antioxidant, analgesic, antistress, and immunotropic. The regulatory mechanism of the action of the substances obtained is suggested.


[Coxarthrosis: clinical aspects, diagnosis and treatment]. Based on a large clinical material the authors analyze the causes of the development of secondary coxarthrosis, suggest new methods of the surgical treatment of the disease, worked out at the Pathology Clinic of Large Joints, Prof. M. I. Sitenko Kharkov Research Institute of Orthopedics and Traumatology, with regard to the shortcomings of the common treatment methods. The term "hyperplastic coxarthrosis" is introduced, the treatment method for that pathology is offered. The problem of further studies and of the design of new methods for the treatment of chronic nonspecific coxitis is under discussion.


[Lumbar spondylarthrosis].

[Lumbar spondylarthrosis]. The results are presented of pathomorphological studies and clinical examinations of 572 patients with lumbar zygo-apophyseal joint arthrosis using arthrography, intraarticular treatment- and- diagnostic blockades, intracutaneous joint denervation and surgical treatment of arthrogenous lumbar canal stenosis. Based on the data obtained, the pathogenetic classification of lumbar spondyloarthrosis has been elaborated and presented in the given paper.


[Angiography in the differential diagnosis of mono-oligo-arthritides in children and adults]. The results of the use of angiography in the diagnosis of chronic mono-oligoarthritides are summarized. The author provides the data on 113 patients aged 2 to 50 years, in whom the diagnosis was confirmed histologically. Describes the angiographic appearance of synovial sarcoma, pigmented villonodular synovitis, juvenile chronic arthritis, juvenile rheumatoid arthritis, tuberculous gonitis, chondromatosis, chondroblastomas, post-traumatic synovitis, hemangiomas, and reactive synovitis in the articular form of osteoid osteoma. The use of angiography improved the radiodiagnosis of mono-oligoarthritides by 45.8%. The conclusion is made that in accordance with indications, angiography may be used as a modality in the system of all-round examination of patients suffering from chronic mono-oligoarthritides.


[Contact radiotherapy of rheumatoid arthritis]. The authors have devised a method of contact radiotherapy with open radionuclides (198Au) to be used in the treatment of patients suffering from rheumatoid arthritis. Indications for therapy have been defined. In 360 patients suffering from rheumatoid arthritis, radioactive colloid gold was administered to the affected joints. The therapeutic effect was slow in development. The best clinical results were recorded at month 12. Considerable improvement was noticed in 70% of the patients, improvement in 20%, and in 10% of the patients, no therapeutic effect could be recorded. Good effect was ascertained in patients with X-ray Stage I and II disease (70%).


[Joint prosthetics using carbon implants]. A group of "Intost" materials based on carbon were developed by the I. M. Sechenov Moscow Medical Academy jointly with the Research Institute of Graphite. After experimental studies the materials were authorized for use in clinical practice for implantation. A concept of the design of endoprostheses of the joints from "Intost" was developed, on the basis of which unipolar combined endoprostheses of the hip joint (127 cases), metatarsophalangeal articulation (27 cases), humeral and metacarpophalangeal articulations (two cases each), and condyles of the tibia (5 cases) were made and applied in clinical practice. A detailed analysis is given of ten years of experience gained with the use of the combined unipolar endoprosthesis of the hip joint manufactured from "Intost-1" and "Intost-3". Long-term beneficial results were obtained in 95% of patients. Endoprosthesis is recommended for the treatment of fractures and false joints of the neck of the femur in elderly and senile patients.


[Restoration of movement in the joints with the aid of a hinge-distraction apparatus]. The device described presents an external hinge which is fixed by transosseous wires and reproduces as far as possible the normal kinematics of the joint. By removing all load from the joint surfaces and maintaining a constant predetermined space between them, the apparatus eliminates friction, prevents pain and muscle spasm, and minimizes abnormal mobility (or instability). It also permits gradual and controlled reduction of old traumatic or congenital dislocations or contractures. Once the bones are in proper relationship, the apparatus permits graduated, controlled, nearly frictionless motion about the correct axis so that a good range of motion is restored and joint surfaces of the proper shape are formed. Finally, ununited fractures close to the joint surface may be immobilized while joint motion is permitted. Experience gained with the apparatus has shown that the method described is physiological and not difficult, complications associated with its use being rare and not serious. The good results obtained with the use of the apparatus warrant its wider application.


[Recombination (regroupings), an important mechanism of quantitative changes in live systems]. In natural phenomena a new quality may occur not only in the course of qualitative changes but also by means of different recombinations (regroupings) of the elements that constitute a system, without any change in their number. Different versions of such regroupings are discussed. Recombination (conformation) changes, widely spread in live nature in particular, are of paramount importance, playing a role in the realization of a wide range of adaptation reactions of the body.


[Changes in creatine kinase activity and protein spectrum of the developing muscle culture treated with creatine phosphate]. A study was made of the influence of creatine phosphate (CP) on creatine kinase activity and the total protein status of the primary chicken myoblast culture. It has been established that the two-hour exposure of the developing culture in the presence of 2 and 4 mM CP is accompanied by the lowering of the content of water extractable protein and a considerable increase of creatine kinase activity. Meanwhile the total activity of the enzyme rises in the non-differentiated and falls in the differentiated one. Electrophoretic analysis has shown CP to produce not only quantitative but also deep qualitative changes in the myoblast protein spectrum. It is assumed that the changes revealed are nonspecific.


[Systemic concepts of real complexity of the organization of the joints]. Cognitive problems arising in arthrology due to discrepancies of analytical means of high complexity as regards the object under examination are analyzed. The use of systemic analysis opens up possibilities of overcoming these difficulties since it permits one (a) to understand specific conformities to natural laws caused by the integrity of the locomotor apparatus and its joints organization, thus affording one to reveal connections between the structure, metabolism, blood supply and joint function in health and disease; (b) to overcome difficulties in interdisciplinary mutual understanding and realization of various knowledge integration on the basis of systemic simulation of the integral object; (c) to arm arthrology with the methodology of task force and goal-oriented planning and organization of multidisciplinary all-round investigations on the basis of integral vision and theoretical analysis of large-scale problems.


Synthesis and secretion by mouse 3T3 cells of an inhibitor of cell growth (mIGFBP-3): correlation with cell proliferation. Our results show that stimulation by serum of dense cultures of 3T3 cells rapidly induced increased synthesis of a growth inhibitor (mIGFBP-3) capable of binding IGF. mIGFBP-3 was secreted by stimulated cells immediately after its synthesis, and accumulated in the medium. Accumulation of mIGFBP-3 in the medium increased, as a function of growth factor (bFGF, PDGF, insulin) concentrations and time. bFGF was the best stimulatory factor for both DNA synthesis and accumulation of mIGFBP-3 in the first 24 h of incubation. DNA synthesis was arrested after 48 h of incubation with bFGF when accumulation of mIGFBP-3 was maximal. Since we showed that mIGFBP-3 is able to inhibit bFGF stimulation of DNA synthesis in mouse fibroblasts, it is possible that the accumulation of mIGFBP-3 induces a feedback regulation of cell growth.


Changes in the ribonucleoprotein constituents of the nucleus during the differentiation of muscle cells in the chick embryo. The ribonucleoprotein components of the nucleus of chick embryo muscle cells in different stages of development were studied by electron microscopic quantitative stereology. The changes of the constituents were related with the appearance of the innervation by means of silver impregnation for light microscope. The numerical density of the perichromatin granules (PCG) is low in mononuclear cells and myotubes. It is noteworthy that the frequency of the PCG does not change during the transition of the cells in mitotic cycle to postmitotic myoblasts and during myofibril differentiation. However, there is an important increment in this parameter when the motor nerve fibers arrive at the muscle and the synaptic contacts are established. This change is correlated with appearance, or at least with a great increase, of the importance of posttranscriptional controls of the expression of some genes. The augmentation in the frequency of PCG is not accompanied by alterations of the abundance of total RNP particles, in close resemblance with the phenomena occurring in neuroblast during the differentiation of synaptic endings. The variations of the nucleolar volume coincide with the changes in rRNA synthesis.


Studies of the molecular biology of parathyroid hormone-like peptide. Molecular biology techniques have been used to examine alternative splicing of the PLP gene, interspecies homology of PLP, the genetic relationship of PLP and PTH. Molecular biology approaches can also be successfully employed to examine both the regulation of PLP production and its function both in emulating PTH and in non PTH-related actions.


Cystatin C based peptidyl diazomethanes as cysteine proteinase inhibitors: influence of the peptidyl chain length. The peptidyl diazomethanes Cbz-Gly-CHN2, Boc-Val-Gly-CHN2, H-Leu-Val-Gly-CHN2, Cbz-Leu-Val-Gly-CHN2 and Cbz-Arg-Leu-Val-Gly-CHN2, with peptidyl portions modelled after the proposed cysteine proteinase interacting N-terminal segment of human cystatin C, were synthesized. Their efficiency as cysteine proteinase inhibitors was tested against papain, human cathepsin B and bovine cathepsin B. All, except Cbz-Gly-CHN2, were found to be irreversible inhibitors of the tested enzymes. Each addition of an amino acid residue to their peptidyl portions resulted in an increased inhibition rate of all three enzymes. These data suggest that the arginyl residue of the tetrapeptidyl diazomethane, and also the corresponding arginyl residue in native cystatin C, interact with a S4 substrate pocket subsite of both papain and cathepsin B. The most efficient inhibitor, Cbz-Arg-Leu-Val-Gly-CHN2, inhibited papain and cathepsin B with rate constants of the same order of magnitude as those for L-3-carboxy-trans-2,3-epoxypropionyl-leucylamido-(4-guanidin o)butane (E-64). The high water-solubility of Cbz-Arg-Leu-Val-Gly-CHN2 allowing it to be dissolved to molar concentrations without use of non-physiological additives, makes it suitable for in vitro and in vivo cysteine proteinase inhibition studies.


Beta-lactams: a new class of conformationally-rigid inhibitors of gamma-aminobutyric acid aminotransferase. A structural similarity of several monobactams (2-4), 3-aminonocardicinic acid (6), 6-aminopenicillanic acid (7), 7-aminocephalosporanic acid (8), and 7-aminodesacetoxycephalosporanic acids (9, 10) to gamma-aminobutyric acid (GABA) and to known inhibitors and substrates of GABA aminotransferase is described. Because of this, the above-mentioned compounds were tested as competitive inhibitors and as inactivators of pig brain GABA aminotransferase. All of the compounds were competitive inhibitors of GABA aminotransferase. On the basis of the inhibitory potency of these conformationally-rigid GABA analogues it is hypothesized that GABA is bound at the active site with its amino and carboxylate groups in a syn orientation. None of the compounds inactivates GABA aminotransferase. These beta-lactam analogues represent the first examples of a new class of inhibitors of GABA aminotransferase.


Inhibition of human alpha-, beta- and gamma-thrombin by mono-, bis-, tris- and tetra-benzamidine structures: thermodynamic study. The inhibitory effect of mono-, bis-, tris- and tetra-benzamidine structures (benzamidine, DAPP, TAPB and TAPP, respectively) on the catalytic properties of human alpha-, beta- and gamma-thrombin (alpha-, beta- and gamma-thrombin, respectively) was investigated (between pH 2.0 and 7.0, I = 0.1 M; T = 37.0 +/- 0.5 degrees C). The affinity of DAPP, TAPB and TAPP for alpha- and beta-thrombin is higher than that found for benzamidine association around neutrality, converging in the acidic pH limb; in contrast, benzamidine, DAPP, TAPB and TAPP show the same value of the association inhibition constant (Ki; M-1) for gamma-thrombin over the whole pH range explored. On lowering the pH from 5.5 to 3.0, the decrease in affinity for benzamidine binding to alpha-, beta- and gamma-thrombin, as well as for DAPP, TAPB and TAPP association to gamma-thrombin reflects the acidic-pK shift, upon inhibitor binding of a single ionizing group. On the other hand, values of Ki for DAPP, TAPB and TAPP binding to alpha- and beta-thrombin appear to be modulated by the acidic-pK shift, upon inhibitor association, of two equivalent proton-binding residues over the same pH range. By considering molecular models of the serine proteinase:inhibitor complexes, the observed binding behaviour of benzamidine, DAPP, TAPB and TAPP to alpha-, beta- and gamma-thrombin has been related to the inferred stereochemistry of the enzyme:inhibitor contact region(s).


Effects of 4-hydroxyandrost-4-ene-3,17-dione and its metabolites on 5 alpha-reductase activity and the androgen receptor. The steroidal aromatase inhibitor, 4-hydroxyandrost-4-ene-3,17-dione (4OHA) and its metabolites, 4-hydroxytestosterone (4OHT), 3 beta,17-dihydroxy-5 alpha-androstan-4-one (metabolite A) and 3 alpha, 17-dihydroxy-5 beta-androstan-4-one (metabolite B) were evaluated as inhibitors of the human prostatic 5 alpha-reductase enzyme and for binding to the rat prostatic androgen receptor. 4OHA and 4OHT were weak inhibitors of 5 alpha-reductase with IC50 values of 15-29 microM. Metabolites A and B had no significant inhibitory activity. 4OHA and metabolites A and B bound weakly to the androgen receptor. The binding affinities (RBA) relative to mibolerone (RBA = 100) were 0.085, 0.485 and 0.016, respectively. However, 4OHT (RBA = 75) was a more potent binder than the endogenous androgen 5 alpha-dihydrotestosterone (RBA = 66). The ability of these metabolites, in particular 4OHT, to bind to the androgen receptor may explain the in vivo androgenic activity of 4OHA.


Irreversible inhibition of thymidylate synthase by pyridoxine (B6) analogues. Three vitamin B6 analogues have been synthesized and tested as inhibitors of thymidylate synthase. The compounds are: 4',5'-dichloro-, 4',5'-dibromo- and 4',5'-diiodo-pyridoxine. All three analogues inhibited the enzyme irreversibly. The kinetic data for the chloro- and bromo-analogues showed that a limiting rate of inhibition is approached as the inhibitor concentration is increased, which indicates that a reversible enzyme:inhibitor affinity complex is formed prior to the irreversible reaction. 4',5'-Dibromo-pyridoxine exhibited a greater binding affinity (lower Ki) for thymidylate synthase than 4',5'-dichloro-pyridoxine, and it also reacted faster to irreversibly inhibit the enzyme. The presence of the substrate dUMP (10 microM) completely protected thymidylate synthase from inhibition. These data suggest that the halogenated vitamin B6 analogues are active site-directed inhibitors of thymidylate synthase, which first bind reversibly to the catalytic site and then react irreversibly with the enzyme.


5838-DNI, a deoxyribonuclease inhibitor produced by Streptomyces sp. strain no. A-5838. 5838-DNI, an inhibitor of deoxyribonuclease (DNase) II from porcine spleen was produced by Streptomyces sp. strain No. A-5838. The structure of 5838-DNI was shown to be 1,4,4a,5,12,12a-hexahydro-4,4a,11,12a-tetrahydroxy-3,8-dimethoxy-9- methoxycarbonyl-10-methyl-1,5,12-trioxo naphthacene. Although similar in structure to tetracenomycin C, which is an antibiotic against Gram-positive bacteria, 5838-DNI has different antibacterial activity. 5838-DNI was distinguished from 5923-DNI, a previously reported DNase II inhibitor, in inhibitory activity against each enzyme. 5838-DNI showed dependency of inhibition on pH and temperature, and inhibited phosphodiesterase I in a competitive manner. These data suggest that 5838-DNI is the first reported example of an inhibitor of microbial origin which is able to inhibit DNase II and phosphodiesterase I.


Novel inhibitors of enkephalin-degrading enzymes. IV: Structure-activity relationships within the penicillins as enkephalinase inhibitors. A range of penicillins have been examined as competitive reversible inhibitors of enkephalinase (neutral endopeptidase; EC 3.4.24.11). Carfecillin (Ki = 0.18 microM) was the most potent inhibitor in the series, whereas cloxacillin (27.5 microM), ampicillin (41.0 microM), nafcillin (58.7 microM) and carbenicillin (158 microM) had moderate potency and benzyl penicillin (885 microM), mezlocillin (473 microM) and azlocillin (556 microM) were weak inhibitors. Structure-activity relationships within the series have been rationalised from a consideration of molecular graphics analysis of the match between receptor binding groups with thiorphan as well as log P values.


Purification and characterization of invertase inhibitors from Dioscorea rotundata tuber. Three invertase inhibitors (A), (B) and (C) from Dioscorea rotundata tuber were resolved on DEAE-cellulose ion exchanger. Two of the inhibitors, (B) and (C), were proteins and homogenous on polyacrylamide gels Mr 21,000 +/- 85 and 26,982 +/- 40/36,307 +/- 50 respectively. The inhibitors (B) and (C) were inactivated at 60 degrees C and had activity-pH optima at 5.2 and 6.4 respectively. (B) and (C) were non competitive inhibitors of invertase from yam and other sources.


Involvement of nitric oxide in spinally mediated hyperalgesia in the mouse. Intrathecal injection of N-methyl-D-aspartate (NMDA) induces a short duration hyperalgesia in mice. An inhibitor of nitric oxide synthase (NOS), N omega-nitro-L-arginine methyl ester (L-NAME), administered either systemically or intrathecally, blocked the NMDA-induced hyperalgesia. This effect was partially reversed by the NOS substrate, L-arginine. Intrathecal hemoglobin mimicked the effects of L-NAME. Intrathecal injection of the NO-donating compounds, sodium nitroprusside (SNP) and hydroxylamine, resulted in a hyperalgesia that lasted 3 h and was reduced by coadministration of hemoglobin. Thus, nitric oxide production appears to mediate NMDA-induced hypersensitivity and may contribute to other forms of centrally induced hyperalgesia.


Neonatal nerve growth factor treatment alters the preganglionic innervation pattern of rat superior cervical ganglion. We treated rat pups with nerve growth factor (10 micrograms/animal/day s.c.) over postnatal days 1-7. Subsequent adult neuron numbers and tyrosine hydroxylase content in superior cervical ganglion were normal, but preganglionic inputs, as gauged from ganglionic choline acetyltransferase, were reduced. In parallel, intraganglionic axon terminals containing calcitonin gene-related peptide, but not those containing substance P, were increased in number. We postulate that neonatal nerve growth factor stimulates sprouting of ingrowing axons that have entered the ganglion soon after birth and that this represses subsequent establishment of cholinergic preganglionic synapses.


Projections and chemical coding of neurons with immunoreactivity for nitric oxide synthase in the guinea-pig small intestine. The distribution of nitric oxide synthase (NOS) immunoreactivity was investigated in the guinea-pig small intestine. There were many immunoreactive nerve cell bodies in the myenteric plexus but very few in submucous ganglia. NOS immunoreactivity was not found in non-neuronal cells except for rare mucosal endocrine cells. Abundant immunoreactive nerve fibres in both myenteric and submucous ganglia, and in the circular muscle, arose from myenteric nerve cells whose axons projected anally along the intestine. NOS immunoreactivity coexisted with VIP-immunoreactivity, but not with substance P immunoreactivity. We conclude that nitric oxide synthase is located in a sub-population of enteric neurons, amongst which are inhibitory motor neurons that supply the circular muscle layer.


Ultrastructural studies of medullary synaptic inputs to vasopressin-immunoreactive neurons in the supraoptic nucleus of the rat hypothalamus. Direct projections from the A1/C1 catecholaminergic cell group in the caudal ventrolateral medulla oblongata to neurons containing vasopressin (VP) in the supraoptic nucleus (SON) were studied electron microscopically by a double-labeling technique which combines anterograde tracing after injection of wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP) into the A1/C1 cell group with VP immunocytochemistry. WGA-HRP-labeled axon terminals originating from neurons in the A1/C1 cell group were found to make synaptic contacts with VP-immunoreactive cell bodies and dendrites in the SON, most often forming axo-dendritic synapses. This indicates that VP-containing neurosecretory neurons in the SON receive monosynaptic catecholaminergic input from the A1/C1 cell group.


Restricted origin and distribution of projections from the lateral to the medial septal complex in rat and guinea pig. The origin and distribution of projections from the lateral to the medial septal complex were studied at the light- and electron-microscopical level in the rat and the guinea pig, with the use of sensitive anterograde tracing techniques. Injections in the lateral septal complex resulted in only weak to moderate labeling in the medial septal nucleus. In contrast, injections in all but the ventral subdivision of the lateral septal complex labeled restricted terminal arborizations in the angular portions of the vertical diagonal band. Analyses at the electron-microscopical level indicated that these fibers form both symmetrical and asymmetrical synapses with dendrites and somata, but not with axons. These findings are discussed in the light of a presumed functional circuit from the hippocampal formation, via the lateral septal complex, to cells in the medial septal complex that originate projections to the hippocampal formation.


Phosphorylation sites on tau by tau protein kinase I, a bovine derived kinase generating an epitope of paired helical filaments. Tau protein kinase I (TPKI) isolated from bovine brain has been determined to phosphorylate tau at four distinct sites by detecting modified Ser and Thr residues with protein sequencer. Ser199, Thr231, Ser396 and Ser413 were all found to have been phosphorylated by TPKI (numbering of amino acids was done in relation to the longest human tau [Neuron, 3 (1989) 519-526]). These phosphorylations generate an epitope of PHF (paired helical filaments) and eliminate the recognition of tau by the monoclonal antibody, tau-1. These results suggested that TPKI might be responsible for at least some of the phosphorylation of tau to induce PHF formation.


Met-enkephalin-Arg6-Gly7-Leu8- and substance P-containing projections from the nucleus preopticus medianus to the paraventricular hypothalamic nucleus. The nucleus preopticus medianus (POMe) is known to be important for the regulation of fluid balance and cardiovascular control. Direct projections from the POMe to the paraventricular hypothalamic nucleus (PVN), where vasopressin-containing neurons exist, were examined in the rat using immunohistochemistry combined with a retrograde tract tracing method. After injection of WGA-HRP-colloidal gold into the PVN, many neurons were retrogradely labeled in the POMe; some of them were immunoreactive to Met-enkephalin-Arg6-Gly7-Leu8 (mE8) or substance P (SP). The results indicate that mE8- and SP-immunoreactive neurons in the POMe send their axons to the PVN.


Apparent anterograde transport of basic fibroblast growth factor in the rat nigrostriatal dopamine system. Five hours after the stereotaxic injection of approximately 1 ng of [125I]basic fibroblast growth factor (bFGF) into rat substantia nigra (SN), small but consistent amounts of radioactivity were found in the ipsilateral striatum (CP). None was found in the ipsilateral thalamus or contralateral CP, SN or thalamus. The amount found in the ipsilateral CP was very markedly reduced in rats injected 5-7 days after 6-hydroxydopamine-induced lesions of the nigrostriatal dopaminergic tract but was unaffected by prior kainic acid lesion of the CP. No radioactivity was found in the ipsilateral SN 5 h after injections of similar amounts of radioactive bFGF into the CP. These results are consistent with anterograde transport of bFGF in the dopaminergic nigrostriatal tract.


Nitric oxide synthase and GABA colocalize in lamina II of rat spinal cord. A dense plexus of fibers in the substantia gelatinosa contains nitric oxide synthase (NOS). Using electron microscopic double-labeling immunocytochemistry for NOS combined with GABA or glutamate, we find that all NOS-positive terminals in this region also contain GABA but are not enriched in glutamate. In an attempt to verify that NOS-positive terminals do not originate from primary afferents, we combined NOS immunocytochemistry with anterograde tracing from the sciatic nerve. An intrinsic spinal origin for the NOS-positive plexus is suggested. The results are discussed in the light of the possible involvement of nitric oxide in hyperalgesia.


Histochemical demonstration of NADPH-diaphorase activity, a marker for nitric oxide synthase, in neurons of the rat pancreas. To clarify the role of nitric oxide (NO) in the pancreas, we histochemically investigated NADPH-diaphorase, a marker for NO synthase, in the rat pancreas. NADPH-diaphorase activity was localized in the neurons and endothelium of vessels. The nerve fibers with NADPH-diaphorase activity were observed in both the exocrine and the endocrine pancreas associated chiefly with the vascular systems. Most nerve cell bodies (98.5%) within the pancreatic parenchyma showed strong activity for this enzyme. These results suggest that NO may participate in the hemodynamic control of the pancreas and in the neuronal regulation of pancreatic exocrine and endocrine functions.


Laryngeal hyperplastic lesions, follow-up study and application of lectins and anticytokeratins for their evaluation. A retrospective study of 878 biopsy specimens from 692 patients with laryngeal hyperplastic aberrations was performed according to the Kambic-Lenart classification. Special attention was focused on 88 patients with persistent or recurring disease. In these carcinoma developed in 17 (2.4%) patients, 12 (1.7%) of whom had had atypical hyperplasia. We therefore propose that the term precancerosis, which so definitely implies cancer, should be replaced with the expression risky epithelium where nothing is determined in advance, but a careful follow-up of the patients is imperative. In particular cases of laryngeal hyperplastic lesions, mainly in abnormal and in atypical hyperplasias when the tissue specimens are cut tangentially, the exact identification and position of individual epithelial cells is essential. In such cases histochemical and immunohistochemical methods yield more precise evaluation. Lectins and cytokeratins provide good markers of epithelial maturation. These results contribute to a more useful evaluation of laryngeal hyperplastic lesions, crucial for the choice of adequate therapy.


Tenascin in reactive lymph nodes and in malignant lymphomas. Tenascin is an extracellular matrix protein which accumulates in the stroma of various malignant and some benign neoplasms. This has been verified in several immunohistochemical studies. The distribution of tenascin immunoreactivity in lymphatic tissues and neoplasias, however, has not been thoroughly studied. In this investigation we analyzed tenascin immunoreactivity in several benign and malignant lymphatic lesions, including both Hodgkin's and non-Hodgkin's lymphomas. In benign lymph nodes, faint reticular immunoreactivity could be observed in the lymphatic tissue. In benign reactive hyperplasias, a stronger reticular pattern of tenascin immunoreactivity was observed in the interfollicular and medullary areas, while the lymphoid follicles contained only a few positive fibers. A similar immunoreactivity was observed in malignant follicular lymphomas. In diffuse lymphomas, a diffuse meshwork of positively stained fibers was seen. This was also the case for the three cases of Hodgkin's disease of the lymphocyte-predominance nodular subtype. There was no difference in the intensity of the immunoreactivity between benign and malignant disorders. However, in Hodgkin's disease of the nodular sclerosis and lymphocyte-depletion subtypes, a much more pronounced immunoreactivity could be observed in the fibrous septa and the cords. This suggests that the tumor cells are possibly capable of synthesizing growth factors which stimulate fibroblasts to synthesize tenascin. The results indicate that tenascin does not accumulate in the stroma of malignant lymphoid neoplasms with the exclusion of some subtypes of Hodgkin's disease. The distribution of tenascin immunoreactivity in lymphatic tissue is similar to that of the reticular fibers suggesting that the molecules are associated with these structures.(ABSTRACT TRUNCATED AT 250 WORDS)


[Pharmacokinetics of procainamide and its metabolite depending on acetylator phenotype]. Procainamide (Pa) and its active metabolite--N-acetylprocainamide (NAPA)--pharmacokinetics was studied in 12 healthy volunteers in relation to acetylation phenotype. Acetylation phenotype was determined with sulphadimidine test. Blood serum levels of PA and NAPA were determined 0.5; 1.0; 1.5; 2.0; 3.0; 4.0; 8.0; and 12 hours following a single oral dose of 500 mg. Blood levels of both PA and NAPA were assayed with immunofluorescence polarization technique (FPIA), using TDx apparatus manufactured by Abbott. Pharmacokinetic parameters were calculated with the aid of pharmacokinetics independent of the model principles. All results were analysed statistically (AWOA). It was found that PA and NAPA pharmacokinetics depends on acetylation phenotype. Blood serum PA levels were higher in slow acetylators during the whole follow-up whereas NAPA levels were lower. Blood serum PA levels in rapid acetylators were decreased while NAPA levels were increased. Acetylation phenotype determined in sulphadimidine test confirmed bimodal procainamide acetylation.


Effects of testosterone, and testosterone plus estrogen, in the castrated rat ventral prostate--histopathological and immunocytochemical studies. The effects of testosterone (T) and 17 beta-estradiol (E2) on the prostate of castrated rats were investigated by histopathological and immunocytochemical procedures. A significant increase in prostatic weight occurred after 6 weeks treatment with T alone and in combination with E2. The greatest increase in prostatic weight occurred after the administration of T plus E2. Histopathologically, glandular hyperplasia of the prostate was noted, and the number of bromodeoxyuridine (BrdU)-positive cells showed a significant increase over that induced by testosterone alone.


[A differential approach to the surgical treatment of liver hemangiomas]. Treatment results and indications for surgery were analysed in 50 cases of hepatic hemangioma versus pattern of hemangiomatosis and size, site and extent of tumor. Surgery was performed in 41 (82%) patients. Radical surgery was possible and justified in as few as one-third of them. Palliative treatment including ligation and roentgen-endovascular occlusion of the hepatic artery and its branches, local ferromagnetic embolization of tumor and suturing and sclerotherapy of hemangioma were performed in 28 cases. End results were analysed in 33 cases followed up to as long as 8 years posttreatment. The extent and type of surgery for hepatic hemangioma should be different for each of the four patterns of the disease identified.


[Palliative operations in pancreatic cancer]. A procedure of transcutaneous draining of bile ducts through a guided replaceable drain using the gallbladder and formation of cholecystostomy and cholecystohepatico-, gastro entero- and enteroenteroanastomoses is suggested for palliation of pancreatic cancer patients. The procedure offers a good clinical effect assuring a significantly longer survival as compared to standard palliative operations used for pancreatic cancer treatment.


[The treatment of nonorganic neoplasms of the lesser pelvis]. In 1967-1989, 330 patients with non-visceral tumors of the small pelvis (children--110 and adults--220) were registered. One hundred and five patients, 17 children included, presented with benign tumors whereas 225 (children--93 and adults--132)--with cancer. Absolute and relative, local and general contraindications to surgery were outlined. The most important criteria are the extent of tumor, patient's status, equipment and surgical skills. Tumors were approached through 9 different accesses depending on their size and site. Surgery wat performed in 242 patients, viz. 89.5% of patients with benign tumors and 65.7% of those with cancer. The percentage of operated children (37.6%) was much lower than that of adults (85.6%) since the former presented with more advanced tumors. Radical surgery was performed in 192 (79.3%) patients; benign tumors were removed in 86.7% whereas cancer--in 44.9%. Complications were encountered in 56.2% of cases, bleeding being the most frequent one (47.0%). Lethality following radical surgery was 3.1%. Radiation and chemotherapy alone proved ineffective. Complex treatment assured better clinical course. One-ten-year survival rate in adults with malignant tumors was 51.2%-31.8%. As few as 17.2% of children survived one year whereas only 6.5%-2 years which points to more aggressive clinical course of malignant non-visceral tumors of the small pelvis in children.


[The characteristics of the combined and complex treatment of colloid rectal cancer]. The effectiveness of combined (surgery + radio- or chemotherapy) or complex treatment for colloidal cancer of the rectum was evaluated in 180 patients with such tumors which included mucinous adenocarcinoma, mucinous cancer and signet-ring cell cancer. Five-year survival rate in 49 patients treated with adjuvant chemo- and/or radiotherapy (39.9 +/- 9.1%) did not differ significantly from that in cases undergoing surgery alone (39.1 +/- 4.7%) (p > 0.05).


[The expression of cytokeratin No. 17 in squamous cell cancer of the cervix uteri: an immunohistochemical study of 19 cases]. Monoclonal antibodies E3 against cytokeratin No. 17 were used for immunohistochemical examination of samples from 19 squamous-cell carcinomas of the uterine cervix and 10 cases of squamous-cell metaplasia of the endocervix. The above antibodies selectively labelled reserve cells of the cervical canal mucosa, basal layer of mature metaplastic epithelium and parabasal and basal cells of immature metaplastic squamous epithelium. In large-cell keratinizing, non-keratinizing and small-cell carcinomas, the expression of cytokeratin No. 17 revealed tendency to basal orientation and was in many respects similar to that in metaplastic squamous epithelium. It was suggested that all the squamous-cell carcinomas studied originated from reserve (metaplastic) cells.


[Familial polymorph ventricular extrasystole associated with Pierre Robin syndrome]. In a family of 9 persons over 3 generations, 6 had incessant polymorphic ventricular extrasystoles, often in salves, resembling unsustained bidirectional ventricular tachycardia. Ventricular repolarisation was abnormal with giant U waves. Five persons had suffered syncopes. All had a facial abnormality: one woman had a complete Pierre Robin syndrome with mandibular hypoplasia, glossoptos and cleft palate; in the other cases, minor forms were observed, with micrognathia and ogival palate. Two patients had bone abnormalities of the feet with total agenesis of the last 2 phalanges of several toes. This familial polymorphic ventricular extrasystole with repolarisation abnormalities has many features in common with the congenital long QT syndrome; associated with the Pierre Robin syndrome and bone abnormalities of the extremities it would seem to be a new multiple congenital abnormalities syndrome.