Human Biopsy Material Research Articles

Giardia lamblia: Electrophysiology and ultrastructure of cytopathology in cultured epithelial cells

The pathogenicity of Giardia lamblia is a subject of debate. Some studies of human biopsy material have mentioned the presence of trophozoites inside the intestinal mucosa, while in others, flagellates have only been found attached to the epithelium. To study the possible cytopathic effects of G. lamblia cultured under axenic conditions, trophozoites of the human 1/Portland and WB strains were placed in contact with monolayers of Madin Derby Canine Kidney cells, a well characterized cell strain with morphological and functional properties similar to those of a transporting epithelium. After 24 and 48 hr of interaction, the effect of the parasite on epithelial cells was assessed by transmission, scanning, and freeze fracture electron microscopy. In addition, the possible action of living trophozoites and sonicates of G. lamblia on the transepithelial resistance of MDCK monolayers mounted in Ussing chambers was analyzed for periods varying up to 48 hr. The results demonstrate that G. lamblia trophozoites do not invade epithelial monolayers. Furthermore, the parasites fail to produce cytoplasmic changes on target cells and have no effect on transepithelial resistance as judged both electrophysiologically and by the failure to open the occluding junctions that bind together epithelial cells. Damage induced by the parasites to cultured cells was limited to focal distortion or depletion of microvilli at the site of adhesion, which may progress to leave circular areas devoid of microvilli, different from the adhesion marks reported by others for G. muris. Therefore, under the in vitro conditions described here, giardias showed no toxic or invasive effect.

Morphological features of the interaction between mucus and surfactant on the bronchial mucosa.

The secretion layer over the bronchial epithelium was investigated on human biopsy material. In the sol phase of the secretion, phospholipid membranes stretched out or arranged in vesicular structures could regularly be demonstrated. They displayed the same structure as the surfactant material in the alveolar space. It can be inferred therefrom that the surfactant is excreted via the bronchial system. It probably contributes to the stabilization of the sol phase of the mucus and thus influences the sliding capacity of the gel phase. So the surfactant has an important function in maintaining mucociliary clearance at the bronchial mucosa.

Relationship between the myofibrillar ATPase activity of human biopsy material and hemodynamic parameters.

The myofibrillar ATPase activity and pyrophosphate gel electrophoretic pattern of native myosin of fresh human left ventricular papillary muscles were examined in 52 cases of mitral valve replacement. The myofibrillar ATPase activity of hypertrophied myocardium did not differ from that of non-hypertrophied myocardium (mean +/- SD, 36.2 +/- 8.7 vs 31.8 +/- 8.6 nmolPi/mg/min, ns) and there was no significant difference in myofibrillar ATPase activity as a function of left ventricular enddiastolic pressure. Pyrophosphate gel electrophoresis of myosin revealed the presence of two components. It is questionable whether the component of higher electrophoretic mobility (approximately 25-35% in concentration) is identical with rat ventricular myosin VM-1 because an increase in this component seems to correlate with a decrease of myofibrillar ATPase activity, its concentration was significantly higher in the hearts with left ventricular hypertrophy, high enddiastolic pressure, high aortic pressure or low cardiac index. From these results, it is not necessarily clear whether hemodynamic overload in valvular heart diseases can alter left ventricular myofibrillar ATPase activity, but it can be said that the overload influences the concentration of the two components of native myosin revealed by pyrophosphate gel electrophoresis.

Discovery of an hourly rhythm of protein synthesis in human gastric biopsy material

Discovery of an hourly rhythm of protein synthesis in human gastric biopsy material

Arteriolar lesions in human renal biopsy material with special regard to the ultrastructural changes in the basal lamina network of the vascular wall.

The arteriolar changes in renal biopsy samples were studied by light and electron microscopy and immunohistologic observations. Arteriolar hyaline thickening was found to occur in virtually all renal diseases, regardless of whether these were accompanied by hypertension or not. Only amyloidosis and dense deposit glomerulonephritis were accompanied by specific ultrastructural arteriolar changes. The nonspecific "hyalin" was shown ultrastructurally to contain various components: accumulated basement membrane material, fine granular deposit (with filamentous or lipid details), and granulovesicular and threadlike membrane structures. Presumably the material constituting these structures originates partly from the blood and partly from elements of the vascular wall itself.

Epstein-Barr virus transcription in nasopharyngeal carcinoma.

Sequences which encode Epstein-Barr virus (EBV) RNA in nasopharyngeal carcinoma (NPC) tissue have been identified. We utilized human biopsy material directly as well as NPC grown in nude mice. Total RNA was extracted from the tumor material and separated into polyadenylated and nonpolyadenylated fractions by oligodeoxythymidylate-cellulose chromatography. This material was used as template to construct 32P-labeled cDNA. The labeled cDNAs were hybridized to Southern blots of recombinant EBV DNA fragments. Three of the biopsies, F, 49, and 55, contained polyadenylated RNA homologous to the EBV BamHI fragments V and K, and EcoRI-DIJhet. These same fragments encode the most abundant polyribosomal RNAs in latently infected lymphoblastoid cell lines. The sequences which encoded nonpolyadenylated RNA in NPC tumor 49 were more extensive and included BamHI fragments C, V, B, E, and K, and EcoRI fragments DIJhet, E, F, and G1, a result that indicates selective polyadenylation in EBV RNA processing. A fourth biopsy, NPC tumor 18, contained polyadenylated RNA homologous to the BamHI fragments H, B, K, Y, B1, I1, and A and EcoRI fragments F and G2. A similar pattern of transcription was identified in three tumor specimens from nude mice, 4, 5, and 8. Transformation of lymphocytes did not occur after cocultivation in vitro with explants from these nude mice tumors. This transcriptional pattern may represent an activated state of the EBV genome, formerly not detected in tumor tissue, which is analogous to the state of abortive infection identified in induced in vitro cell systems.

A rapid ultrastructural embedding procedure for renal biopsy tissue

The increasing reliance of pathologists on ultrastructural morphology for the diagnosis of human biopsy material has created a need for rapid ultrastructural embedding procedures (RUEP) to accelerate the preparation of material. The reduction of technical time required for complete (wet tissue to stained grid) processing of a biopsy specimen, from the several days [3–5] of conventional tissue processing to less than 1 day, greatly aids the physician in patient management, particularly in renal pathology. Differentiation between lipoid nephrosis, dense deposit disease, membranous glomerulonephritis, and amyloidosis is accomplished easily at the ultrastructural level, and in some cases, can best be diagnosed by electron microscopy. The extent and pattern of distribution of immune complexes in systemic lupus erythematosus determine both the aggressiveness of treatment and the prognosis.

Separation and identification of subpicomole amounts of methotrexate polyglutamates in animal and human biopsy material

A rapid, quantitative procedure for separating methotrexate (MTX) polyglutamates utilizing molecular-sieve column chromatography has been developed. MTX polyglutamates were separated in 25 min and detected by a radioassay with a sensitivity of 0.1 pmol MTX, allowing analysis on 2 to 5 mg of tissue. This technique has been used to demonstrate and quantitate the levels of MTX polyglutamates in liver, kidney, and brain tissues of rats treated with weekly low doses of MTX, and in liver and red cells of patients with acute leukemia treated with weekly low doses of MTX.

Cutaneous Form of Bovine Papular Stomatitis in Man

A cutaneous form of bovine papular stomatitis (BPS) infection was diagnosed in eight persons at the School of Veterinary Medicine at Auburn University, Auburn, Ala. The initial outbreak occurred in five persons who were involved in the care of a bull that required manual placement of an oral feeding tube. Confirmation of diagnosis was based on clinical findings, cytopathological effects in tissue culture, and isolation of typical paravaccinia virus particles in tissue culture. Transmission studies were performed successfully in three normal calves using tissue culture prepared from human biopsy material. In man, the cutaneous form of BPS infection shows gross lesions similar to the cutaneous form of contagious ecthyma ("orf") or pseudocowpox ("milkers' nodules") infection. Because BPS in cattle occurs most often without evidence of readily observable lesions, unlike contagious ecthyma in sheep or pseudocowpox in cattle, the transmission of BPS to man in the cutaneous form could occur without apparent source. The mild clinical manifestations make the condition relatively minor; however, the occasional case may have more severe lesions.

High resolution radioautography of central dopaminergic fibers labelled in vitro with [ 3H]dopamine or [ 3H]norepinephrine

An original method is reported for the radioautographic visualizing of dopaminergic axon terminals in the rat central nervous system. This method is based on an in vitro labelling with either tritiated norepinephrine or dopamine under conditions of specificity previously established by fluorescence histochemistry. The immobilization of [ 3H]catecholamines in their sites of uptake and storage during post-fixation for electron microscopic processing was ensured by the use of osmic acid vapours instead of osmic acid solution. Selective labelling of the dopaminergic fibers may be achieved throughout large brain sections and the techniques should be applicable to human biopsy material.

Immunofluorescence studies on the subcomponents of the first component of complement (C1): Detection of C1q and C1s in different cells of biopsy material and on Human as well as on guinea pig peritoneal macrophages

The first component of complement (C1) ist a macromolecule consisting of three distinct subcomponents, C1q, C1r, and C1s. In regard to its production site and its role in phagocytic processes it was of interest to find out whether these different subcomponents could be detected in human biopsy material only as a complex in individual cells or whether C1 subcomponents could be found on different cells. To study this question, monospecific fluorescein-labelled anti-human-C1q IgG and monospecific rhodamine-labelled anti-human C1s IgG were used. Biopsy material from human rectum was stained with fluoresceinated antisera, either by use of one antiserum or by double staining. Using this technique, these oberservations were made: C1q as well as C1s were detectable in individual cells in the subepithelial area of the gut. Furthermore, C1q and C1s could be found together in the same cell or separately in different cells. These findings were supported by experiments with cultured peritoneal macrophages either from human or from guinea pig. The examination of the cultured cells with the two antisera revealed that individual cells were stained either by anti-C1q or by anti-C1s antibodies. The specificity of the detection of the individual subcomponents was also proven by the peroxidase technique and by using fluoresceinated anti-human C1q F(ab') 2. The membrane immunofluorescent staining revealed the presence of C1q on the membrane of the macrophage.

The significance of in vitro activation of guinea pig complement in glomeruli of human renal biopsy materials from varied subtypes of glomerulonephritis.

To measure the potential ability of complement activation by glomerular-bound immune complex or C3 convertase, the in vitro fixation of guinea pig complement (GPC) components to glomeruli was examined by immunofluorescence on the frozen sections of human renal biopsy materials, from the varied subtypes of glomerulonephritis (GN). Study for the pathway of complement activation in each subtype was also done. The extent of GPC activation fairly well paralleled with those of the autologous immunoreactants depositions and further with the morphological alterations of glomeruli in the cases of proliferative GN, advanced membranoproliferative GN (MPGN), lupus nephritis, purpura nephritis of non-IgA nephritis type and advanced focal segmental glomerular sclerosis. The ability of GPC activation was present but generally weak in the cases of post-streptococcal acute GN (post-str. AGN) and subsiding MPGN. The rate of positivity was further reduced in the cases of membranous nephropathy (MN), IgA nephritis and related purpura nephritis. The dominant alternative pathway of GPC activation was found in MPGN, post-str. AGN, IgA nephritis, and related purpura nephritis, the only classical pathway was noted in MN.

Tooth ankylosis in man and the oxytalan fibre meshwork

Abstract There is no published information on the presence of the oxytalan fibre meshwork in the periodontal ligament of human ankylosed teeth. Contrary to the limited term experimental animal study of Hurst (1972), the present investigation demonstrates that extensive oxytalan structures can persist over many years in the presence of ankylosed teeth. Human biopsy material of four ankylosed first and second permanent molars exhibited a well-defined oxytalan meshwork that linked the teeth and periodontal vessels to each other proving that oxytalan-vascular associations are maintained in the absence of tooth oscillation. Thus, oxytalan fibres are not only found in connective tissues subject to stress (Fullmer et al. 1974), but also in the relatively stress-protected periodontium of submerged teeth. These findings raise new questions regarding the possible functional role of the oxytalan fibre meshwork.

Changes in Morphology and Cellular Replacement of Surface Epithelia in Human Gastric Mucosa Affected by Different Pathological Changes: A Scanning Electron Microscopical Study of Human Gastric Biopsy Material

The surface epithelium of normal gastric mucosa shows a constant turnover, migrating from the proliferation zone at the neck of the gastric glands to the mucosal surface. The cytologic changes associated with the concomitant cellular differentiation were so far studied in sections by light and electron microscopy only. With these methods neither the changes of surface morphology associated with cellular maturation nor the shedding of surface epithelia can be recorded. With the scanning electron microscope it is possible to study especially these changes of the mucosal surface plus those of the single surface epithelia over a wide range of magnification and with a sufficient depth of focus. In this study therefore an attempt was made to correlate some scanning electron microscopic findings concerning these changes of mucosal and cellular surface with the results of parallel light microscopic examinations. For this purpose double biopsies of human gastric mucosa were taken during routine endoscopic examination. One of these biopsies was processed for light microscopic examination after formaldehyde fixation. The other was fixed in buffered glutaraldehyde solution and 1 % OS04 solution and thereafter the material was processed for scanning electron microscopic study by dehydration against an increasing acetone gradient, critical point drying and sputter coating of the dry specimens which were mounted on metal stubs with conductive silver. Some aspects of surface morphology of the mucosa and the single cells in correlation to different types of gastritis are demonstrated and the morphological expressions of cytological degeneration and shedding from the mucosal surface are commented on.

A study on the digestion of human muscle biopsies for trace metal analysis using an organic tissue solubilizer

A commercially available tissue solubilizer, based on tetraalkyl-ammonium hydroxide in toluene, has been compared with conventional digestion producers in order to determine the minimum amount of tissue needed for the determination of trace elements such as Al, Cd, Cu, and Mn in human biopsy material using flameless atomic absorption spectrophotometry. The solubilizer can be recommended for digestion of biopsy samples in order to determine Cu, Cd, and Mn, but not Al.

Ultrastructural observations on giardiasis in a mouse model. II. Endosymbiosis and organelle distribution in Giardia muris and Giardia lamblia.

Ultrastructural observations of Giardia muris in a mouse model revealed endosymbiotic microbes not previously reported in Giardia. Endosymbionts 240--360 nm wide, 600--1,400 nm long, and with an internal structure similar to that of bacilli were not seen entering Giardia but were found and appeared to divide within Giardia. No evidence was found of digestion of the endosymbionts by the giardia host in either the trophozoite or the cyst form. Endosymbionts were concentrated centrally around the nuclear area and were uncommon in peripheral feeding regions. The same cellular organelles seen in G. muris were found in Giardia lamblia from human jejunal biopsy material, but no endosymbionts were identified in G. lamblia trophozoites from the seven patients examined. Endosymbionts within Giardia may be found to alter trophozoite pathogenicity, metabolism, range of infectivity, antigenic surface characteristics, and host specificity, as they do in other protozoa.

Amplified direct immunofluorescence (AMDI) for detection of Epstein-Barr virus nuclear antigen

Low antigen concentrations could be identified in human cells by sequential application of (a) FITC-labeled human antibodies at high dilution, (b) rabbit antiserum to the hapten fluorescein isothiocyanate (FITC), and (c) FITC-anti-rabbit globulin. The high specificity of direct immunofluorescence (a) was not affected by the amplifying steps (b) and (c). Using this AMDI technique Epstein-Barr (EB) virus nuclear antigen (EBNA) could be specifically stained with human sera up to a dilution of 1 : 4000. Owing to the high dilutions applied, unwanted antibody reactivity in the FITC-labeled serum could be blocked by preincubating with unlabeled undiluted human sera. Thus EBNA was selectively stained in EB virus producer cells. Moreover, EBNA was specifically detected in human tumor biopsy material by the use of AMDI.

Healing of the dento-epithelial junction following subgingival plaque control. 1. As observed in human biopsy material : WAERHAUG J. J. Periodontol. 49 (1978) 1–8

Healing of the dento-epithelial junction following subgingival plaque control. 1. As observed in human biopsy material : WAERHAUG J. J. Periodontol. 49 (1978) 1–8

Creutzfeldt-Jakob disease: Do patients have viremia?

Evidence has been obtained for the presence of infectious virus in the blood of guinea pigs with experimental Creutzfeldt-Jakob disease. If these results are replicated in humans with this type of spongiform encephalopathy, it could explain both systemic spread of the disease and raise a warning about transmissibility (through blood transfusions, for instance) during the long, symptomless period of incubation. The work was reported recently by Elias and Laura Manuelidis, both MDs and professors of neurology and pathology at the Yale University School of Medicine. In 1975 these investigators were the first to transmit Creutzfeldt-Jakob disease to laboratory rodents using human biopsy material from patients with the disease. In the current experiments reported in the June 2, 1978, issue of Science , they used brain tissue from guinea pigs that had been inoculated originally with material derived from the brain of a patient with Creutzfeldt-Jakob disease. This guinea-pig brain tissue was

Electron-microscopic studies on primate neurons: microtubule-pigment associations.

The relationship between microtubules and the lipoprotein pigment found in senile Cynomolgus brain and senile human brain biopsy material was investigated. Numerous microtubules were present in all parts of the cytoplasm and within the pigment areas, running parallel or obliquely to the pigment bodies and being associated with its lateral aspects. Microtubules also occurred in the periphery of neurons or appearing to enter the perineuronal oligodendrocytes. These observations indicate a possible role of microtubules in the transport of pigment bodies. The oligodendrocyte from human brain biopsy material has definitely taken the role of a phagocyte in ingesting pigment bodies. Its numerous microtubules may offer a fasting moving system for disposing off pigment residures to the capillary endothelium.