N-acetyl D-galactosamine specific lectins were isolated from the seeds of Jack Fruit (Artocarpus integrifolia) and Winged bean (Psophocarpus tetragonolobus) and D-galactose specific lectin was isolated from peanut (Arachis hypogaea). These lectins were conjugated to Horse Radish Peroxidase (HRP) and were used to study the lectin binding properties of benign and malignant lesions of the thyroid. For comparison of the results 10 normal fresh autopsy specimens were included in the study. The Peanut lectin (PNL) and Jack fruit lectin (JFL) conjugates showed positive binding with the cells in different lesions, while Winged Bean Lectin (WBL), despite its having a common inhibitory sugar, showed no binding even after neuraminidase treatment. These lectins revealed difference in the composition of glycoconjugates of benign and malignant thyroid cells. The HRP conjugated JFL and PNL may be of use in distinguishing carcinomatous tissues from benign tissues which makes them potential tools in the differential diagnosis of thyroid lesions.
The prevalence and titre of IgG antibodies to human herpesvirus type 6 (HHV-6) were assayed in the serum samples from normal subjects and patients with Hodgkin's lymphoma (HL), non-Hodgkin's lymphoma (NHL), acute lymphoblastic leukaemia (ALL) and oral cancer (OC) using immunofluorescence and immunoperoxidase techniques. This forms the first study on the sero-prevalence and titre of antibodies to HHV-6 in India. There was no considerable difference in the prevalence (76%) and titre (10-160) of the antibodies in normal population from those reported for normal adults in other parts of the world. All the HL and ALL patients studied showed no significant elevation in the antibody titre, though a slight increase in the prevalence (95%) was noted. Antibody titre and prevalence were found highly elevated in OC. OC remained totally unstudied for the presence of anti-HHV-6 antibodies, and this is the first report of elevated levels of the antibody in this cancer. The role of HHV-6, if any, in the pathogenesis of OC is worth investigating.
Our data showed that the regenerative capacity of skeletal muscle in newborn guinea pigs was less than in adults. In newborn guinea pigs the active regeneration of muscle tissue was only observed during the 1st week after mincing of gastrocnemius muscle. The 14-day grafts were characterized by having smaller muscle fibers and greater amounts of interstitial connective tissue. Furthermore, the active growth of connective tissue was more pronounced. The grafts did not recover contractility. In 60 day grafts the ectopic formation of 2-3 bone-cartilage nodules was observed. In adult guinea pig grafts the quantity of muscle tissue was twice as great as in newborns. Muscle tissue was formed throughout the graft, but no bone-cartilage nodules were present. All 60 day grafts contracted when the tibial nerve was stimulated, but this contractility was very weak. However, the reestablishment of neuromuscular junctions had occurred. The age characteristics of skeletal muscle regeneration were the same both in the term guinea pig and in the rat.
In our previous study, we reported that monocyte-activation inhibitory factor was produced by stimulated fibroblasts. We also previously found that glycyrrhizin (GL) had an ability to affect fibroblasts because the proliferation of human fibroblasts was increased by GL. In this study, we demonstrated that culture supernatants from the GL-stimulated fibroblasts inhibited the activation of normal human peripheral monocytes in vitro. Then, studies were performed to know whether GL affects fibroblasts to suppress the granuloma formation. Pulmonary granulomas were induced in guinea pigs by Sephadex beads. The formation of granulomas was significantly suppressed by intraperitoneal injections of GL. Thus, GL was shown to have antigranulomatous effects in vivo.
The effects of the vagus and its action on the mucosal mast cells (MMC) of the secretory portion of the rat stomach are analyzed by observing the consequences caused by subdiaphragmatic truncal vagotomy on the MMC in terms of cell count and degranulation over a period of four months. Observations showed a gradual decrease in the number of mast cells/mm2, an increase in the percentage of MMC in a state of degranulation, but the same number of degranulated cells/mm2. This suggests that the vagus controls both numerically and functionally the MMC population in the secretory portion of the rat stomach.
Lectin binding to tumor cells in tissue sections of nonmetastatic and metastatic murine Lewis lung carcinoma (LLC) was assessed by light and electron microscopy using a lectin-gold technique. Ulex europaeus agglutinin-I (UEA-I) and peanut agglutinin (PNA) showed no binding, whereas concanavalin A (Con A), soybean agglutinin (SBA), Dolichos biflorus agglutinin (DBA), Maclura pomifera agglutinin (MPA), and Ricinus communis agglutinin-I (RCA-I) bound equally to the transplanted sites and metastases. However, wheat germ agglutinin (WGA) bound to metastases more highly than to the transplanted sites and there was a statistically significant difference (P less than 0.01) between the transplanted sites and metastases with regard to pre-embedding method. The tumor cells binding to WGA clearly decreased in number after sialic acid pretreatment and were rich in more well-differentiated organelle. In the bromodeoxyuridine (BrdUrd) labeling in vivo, cell proliferation was greater in the metastatic sites than in the transplanted sites. The above findings suggest that glycoconjugates on the tumor cell surface are altered in the process of metastasis and correlate with metastatic potential and cell proliferation.
The neurotoxic cancer chemotherapeutic agent cis-diamminedichloroplatinum (II) (cis-platin) was tested in a model system of cultured embryonic chick dorsal root ganglion cells, in order to investigate cellular mechanisms of toxicity. At 7.5 ug/ml, the drug caused mild toxicity. At doses of 75 ug/ml, cis-platin was toxic to cultures in 6 hours, in both neuronal and non-neuronal cell populations. After 24 hours of incubation with 75 ug/ml cis-platin, there was extensive cell death. The trans isomer of the drug, trans-platin, was less toxic than cis-platin at similar doses, causing less severe damage to the cells as well as less cell death. With both drugs, abnormalities in patterns of nuclear staining were prominent, whereas neuronal cell membrane staining patterns were less affected. Both drugs seemed to affect non-neuronal cells to a greater extent than neurons. Ultrastructural findings with both drugs included nucleolar segregation; mitochondrial changes were nonspecific. In this in vitro system, both cis- and trans-platin are toxic. The toxicity appears to predominantly affect the nucleus, and to preferentially involve non-neuronal cells.
Human serum injected intravenously into rats caused multiple foci of acute enteritis. The enteritis had a predilection for the antimesenteric side of the intestine and for the zones between circumferential vessels. Despite their antimesenteric location, Peyer's patches tended to be spared. The details of distribution suggest that a gradient related to intestinal blood flow plays a role in development of the enteritis.
We examined the effect of type I interferon (IFN) on IFN-gamma-induced HLA-DR antigen expression in A431 cells, a human squamous cell carcinoma line. A431 cells expressed HLA-DR antigen when stimulated with IFN-gamma, but not with IFN-beta. Simultaneous addition of IFN-gamma and IFN-beta to A431 cells resulted in significantly decreased HLA-DR antigen expression when compared to treatment with IFN-gamma alone. Kinetic studies revealed that IFN-beta was required concomitantly or prior to stimulation with IFN-gamma in order to down-regulate expression of HLA-DR antigens. IFN-alpha also inhibited IFN-gamma-induced HLA-DR antigen expression in A431 cells. Analysis of cytoplasmic mRNA showed that simultaneous treatment of A431 cells with IFN-gamma and IFN-beta resulted in a marked decrease of the level of DR alpha specific mRNA when compared to a level reached after treatment with IFN-gamma alone. These results suggest that type I IFN antagonize the IFN-gamma-induced HLA-DR antigen expression in human keratinocyte system, and that this antagonistic effect of type I IFN is confirmed as evidenced by a change in HLA-DR mRNA levels.
Aortic lysosomal enzyme activities have been evaluated in relation to the extent and severity of aortic atherosclerosis in rhesus monkeys to see the biochemical and pathological effects of renal hypertension in experimental atherogenesis. The frequency and size of atherosclerotic plaques in aortas of atherogenic diet fed and/or hypertensive monkeys were calculated and an overall score of aortic atherosclerosis was computed on the basis of the gamut of pathological findings in relation to the biochemical alterations. This overall score of atherosclerosis was found to be significantly (p less than 0.01) greater in animals of all experimental groups as compared to the controls.
The effect of AZT on serum HIV p24 antigen and endogenous serum alpha interferon levels was studied in AIDS and ARC patients. Following administration of AZT there was a rapid decline in the serum levels of both HIV p24 antigen and alpha interferon. When AZT treatment was interrupted, the levels of both HIV p24 antigen and of interferon rapidly increased. These findings suggest that HIV or some other AZT sensitive microorganism is the inducer of interferon which is characteristically found in the serum of AIDS and symptomatic HIV infected patients. They also suggest that the rapid decline in interferon levels may underlie some of the symptomatic benefit that follows administration of AZT.
Whether infection with influenza B virus alters hepatic function was examined in the ferret. Also, the possibility that viral-specific antibodies (Ab) could be produced well before their detection in serum was explored. During the febrile period of influenza, reductions in the serum potassium, anion gap, ammonia, albumin and CPK and elevations of the BUN, creatinine and the GGTP levels occurred. With convalescence, the electrolytes, BUN and creatinine normalized, FFA, SGPT and CPK levels rose and the serum GGTP rose even further. Hepatic fatty acid (FA) oxidation, ornithine transcarbamylase (OTC) and carnitine palmitoyltransferase (CPT) activities were minimally altered and liver ATP and total lipid content remained normal. Following experimental secondary viremia, serum FFA continued to rise, TG decreased and CPK remained elevated while SGPT and GGTP levels normalized. In the liver, FA oxidation and OTC rates remained unchanged but CPT activity was inhibited and the liver content of ATP was significantly reduced. Immune complex (IC) protein recovered from postmicrosomal supernatant fractions by polyethylene glycol precipitation was progressively increased in livers from convalescent and viremic animals. While the amount of IC protein recovered in the spleen also increases during convalescence, this is not the case after viremia when the IC formed seem to be processed largely by the liver. By SDS/PAGE, the major proteins identified in the IC were IgM and other viral proteins. However, the viral proteins could not be validated by immunoblot with Ab produced against purified influenza B hemagglutinin (HA) and neuraminidase (NA) most probably due to phagocytic alterations of glycoprotein immunodeterminants. These findings indicate that during influenza, convalescence and post viremia changes in the concentrations of several serum and liver components occur that reflect hepatic involvement. Also, antiviral Ab, largely IgM, appears to be produced early, complexes with Ag and can be found sequestered in both the liver and spleen at a time when Ab is not detectable in the serum.
Biopsy samples from one hundred and two patients with squamous cell carcinoma of the uterine cervix and tissues from twelve healthy normal cervical tissues (post hysterectomy) were examined for HSV-type 2 and HPV-11 DNA sequences by molecular hybridization technique. In the carcinoma tissue extracts 53% contained HSV-2 DNA, 27% -HSV-1-DNA and 36% showed HPV-11 gene sequences while 5.7% were found to contain both HPV and HSV-2 DNA. Biopsies from healthy cervix were completely negative for HSV-2 and HPV-11 DNA sequences.
Paraffin sections of biopsies from histopathologically confirmed cases of uterine cervical carcinoma, cervical scrapings from dysplasia, chronic cervicitis, tumour cells from carcinoma of the oral cavity and normal tissues from healthy normal cervix and oral cavity scrapings were examined for the presence of Human papilloma virus antigens. The techniques adopted were the Indirect Immunofluorescence Stainig and the Peroxide-Anti-Peroxidase techniques. The HPV-antigen was present in 38 percent and 41 percent of invasive carcinoma cervix, by PAP and IIF methods respectively. In cervical dysplasia 8-13% revealed HPV antigen while oral carcinoma cells and normal tissue samples were totally negative.
MRL1/1 & NZB/WF1 female mice were treated with high-dose cyclophosphamide (20 mg/kg/week), and the distribution of T cells in the Peyer's patches was examined in treated and untreated mice. A multiple layering technique was used for immunohistochemical detection of the lymphocyte surface antigens of T cells (Thy1.2, L3T4, Lyt2). High-dose cyclophosphamide inhibited the increase of the T cell population of MRL1/1 female mice with age, but little change was observed NZB/WF1 female mice.
Treatment of adult mice with gold sodium thiomalate made the normally non-lethal Semliki Forest virus infection lethal. Associated with this was a significant increase in brain virus titre and a depression of peritoneal macrophage lysosomal enzyme activity. In contrast, treatment of adult mice with the constituent part, thiomalate, did not make the non-lethal Semliki Forest virus infection lethal. Brain virus titre and peritoneal macrophage lysosomal enzyme activity were comparable to the controls. The mechanism by which gold sodium thiomalate increases the virulence of Semliki Forest virus is discussed.
Surgical samples of human hepatic tissue were analysed morphologically and biochemically and highly differentiated hepatomas were compared with two control groups: morphologically normal liver tissue surrounding the tumour, and tissue from normal livers. In tumour homogenates cholesterol levels were more than twice, ubiquinone levels about half and the concentration of free dolichol about 10% of the control value. The levels of dolichyl phosphate were basically similar, whereas the phospholipid level was slightly lower in the tumours. In microsomes isolated from hepatomas, the level of cholesterol was about 30% higher than the control value. HMG-CoA reductase activity in microsomes isolated from hepatomas was elevated almost 100% in comparison to control. In hepatomas, no major alterations in the compositions of dolichol or dolichyl phosphate could be observed. The relative amounts of alpha-saturated and alpha-unsaturated polyprenols were also basically unaltered in hepatomas. Liver samples were incubated with 3H-mevalonic acid and radioactivity was monitored in polyprenols. With control tissue, incorporation was considerably higher in alpha-unsaturated polyprenols than in their alpha-saturated counterparts. In the tumours the rates of incorporation into both polyprenol fractions were much lower, although still higher in the alpha-unsaturated fraction. Labelling of polyisoprenols containing 19 isoprene residues was higher than that of 20 residues. The pattern of labelling in the polyisoprenyl-P fraction was similar. In hepatomas the incorporation into cholesterol and ubiquinone-10 was about 100% higher and 50% lower respectively compared with control tissue. The results in this study of hepatomas indicate that the levels of various lipids may be influenced not only by the regulatory enzyme HMG-CoA reductase, but also by other enzymes catalysing reactions subsequent to this regulatory point. It is also suggested that levels of cholesterol, ubiquinone and dolichol may be regulated independently subsequent to the branch point at farnesylpyrophosphate.
The development of left ventricular ischaemic contracture and its correlation with ultrastructural and sarcolemmal permeability defects were studied in isolated rat hearts during global subtotal ischaemia. With acetate as substrate the hearts exhibited a rise in diastolic tension after 8-10 min at which time small foci of contracted myocytes were scattered throughout the myocardium. In hearts with 5% of the maximum diastolic tension (termed 5% contracture), the foci were situated predominantly in the subendocardium and papillary muscle. Contracted myocytes in these foci were capable of excluding ionic lanthanum thus demonstrating retention of normal sarcolemmal permeability properties. With 30% contracture ultrastructural damage had spread to the subepicardium and with further contracture there was an associated increase in the number and size of foci in all regions. In these foci, swelling of the tubular sarcolemmal system and occasionally of the sarcoplasmic reticulum appeared to precede myofibrillar contraction. At 50% contracture lanthanum influx into contracted cells became more frequent. Hearts developed full contracture by 15-18 min at which time most myocytes were contracted and retained lanthanum intracellularly. The heterogeneity of the response at a cellular level may offer a possible explanation for the lack of correlation between contracture and tissue ATP. A possible sequence of structural injury leading to impaired calcium homeostasis is also suggested.
'Stress Proteins in Inflammation' provided a forum for the discussion of topical issues in this rapidly moving field. The mycobacterial 65 kDa stress proteins play a key role in certain animal models of inflammatory arthritis. However, the impression emerging is that the mechanism probably involves more than a simple cross-reaction between mycobacterial SP65 and either the host SP65 or a cartilage antigen, and that evidence for a primary role in human rheumatoid arthritis is lacking. A realistic role for immune responses against stress proteins might be the amplification or perpetuation of inflammation. If so, this is unlikely to be limited to arthritis.
