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Biomedical subjects

K J Kramer

Publications and source records attributed to K J Kramer.

At least 73 records · Page 4Linked to original sources

Plasmodium falciparum: comparative analysis of antigens from continuous in vitro cultured and in vivo derived malarial parasites.

A comparison of metabolically labeled proteins from continuous in vitro and in vivo derived Plasmodium falciparum revealed both similarities and differences. Metabolic labeling of synchronized cultures showed that the uptake of label increased as the parasites matured from the ring to the schizont stage in both cultures. Also, in both continuous in vitro and in vivo derived cultures, prominent high-molecular-weight proteins were synthesized during the late developmental stages. However, the continuous in vitro cultured parasites incorporated twice as much of the label at each stage as did the in vivo derived parasites. Immunoprecipitation with serum samples from vaccinated Aotus trivirgatus griseimembra monkeys revealed major differences involving protein antigens that migrated in the molecular weight regions of b (Mr = 152,000), c (Mr = 143,000), j (Mr = 82,700), and n (Mr = 57,400). These antigens were more readily detected in the continuous in vitro cultured schizonts than in the in vivo derived schizonts.

Animals↗

Immunocytochemical evidence for the occurrence of insulin in the frontal ganglion of a Lepidopteran insect, the tobacco hornworm moth, Manduca sexta L.

The frontal ganglion of the adult forms of the tobacco hornworm, Manduca sexta, was investigated immunocytochemically for the occurrence of the gastro-entero-pancreatic (GEP) neurohormonal peptides, namely insulin, nerve growth factor, epidermal growth factor, insulin C-peptide, somatostatin, glucagon, glicentin, pancreatic polypeptide (PP), polypeptide YY (PYY), secretin, vasoactive intestinal peptide (VIP), gastric inhibitory peptide (GIP), gastrin, cholecystokinin (CCK), enkephalin, alpha- and beta-endorphins, substance P, neurotensin, bombesin, motilin, ACTH, serotonin, and calcitonin. Among all the antisera tested, positive immunostaining was obtained with anti-insulin B-chain serum only. The insulin B-chain immunoreactivity was localized in 4-6 large (30-40 microns) neurons, in the neuropile, and in the recurrent nerve. It is speculated that the insulin-like immunoreactive material may be transported to the neurohaemal organ (corpora cardiaca) through the nervi cardiaco-somatogastrici.

Animals↗

Plasmodium falciparum: protein antigens identified by analysis of serum samples from vaccinated Aotus monkeys.

Serum samples from Aotus trivirgatus subsp. griseimembra monkeys obtained at different stages of a vaccination experiment were analyzed for total antibody titer to Plasmodium falciparum and were used for identifying protective antigens of the human malaria parasite. Total malarial antibody titers were higher in serum samples from protected monkeys (vaccinated with antigen in an adjuvant) than in those from unprotected monkeys (vaccinated with either antigen or adjuvant only). Parasite proteins were labeled with [3H]isoleucine, solubilized with nonionic detergent, and reacted with immune Aotus sera. Immunoprecipitates obtained were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography. Thirteen protein antigen bands in the molecular weight range 73,000 to 180,000 were resolved. Serum samples obtained from protected Aotus monkeys reacted more intensely with these proteins than samples from unprotected monkeys did. Evidence is presented that the protective antigen is not a single, normally nonimmunogenic, protein that is recognized only in protected monkeys. Rather, the present data indicate that a heightened immune response to multiple proteins correlated with in vivo protection to P. falciparum in Aotus monkeys. This finding may have a significant bearing on strategies for the development of a human P. falciparum vaccine.

Animals↗

Immunohistochemical investigations of neuropeptides in the brain, corpora cardiaca, and corpora allata of an adult lepidopteran insect, Manduca sexta (L).

In the brain of adult specimens of the tobacco hornworm moth, Manduca sexta (L), cells immunoreactive for several kinds of neuropeptides were localized by means of the PAP procedure, by use of antisera raised against mammalian hormones or hormonal peptides. In contrast, no such neurosecretory cells were found in the corpora cardiaca and corpora allata (CC/CA); in the CC/CA, however, immunoreactive nerve fibres were observed, reaching these organs from the brain. The neurosecretory cells found in the brain were immunoreactive with at least one of the following mammalian antisera, namely those raised against the insulin B-chain, somatostatin, glucagon C-terminal, glucagon N-terminal, pancreatic polypeptide (PP), secretin, vasoactive intestinal polypeptide (VIP), glucose-dependent insulinotropic peptide (GIP), gastrin C-terminus, enkephalin, alpha- and beta-endorphin, Substance P, and calcitonin. No cells were immunoreactive with antisera specific for detecting neurons containing the insulin A-chain, nerve growth factor, epidermal growth factor, insulin connecting peptide (C-peptide), polypeptide YY (PYY), gastrin mid-portion (sequence 6-13), cholecystokinin (CCK) mid-portion (sequences 9-20 and 9-25), neurotensin C-terminus, bombesin, motilin, ACTH, or serotonin. All the neuropeptide-immunoreactive cells observed emitted nerve fibers passing through the brain to the CC and in some cases also to the CA. In CC these immunoreactive nerve fibers tended to accumulate near the aorta. It was speculated that neuropeptides are released into the circulating haemolymph and act as neurohormones.

Animals↗

Electron microscopic observations on alterations in cultured Plasmodium falciparum and infected erythrocytes after treatment with cyclic AMP in vitro.

The fine structure of Plasmodium falciparum treated with cyclic AMP in vitro was studied. Cyclic AMP stimulated the appearance of membranous structures in P. falciparum-infected erythrocytes. Two types of membranous structures originating from the host cell were observed: multilaminate membranous structures and multistranded layer-like membranous structures. The multilaminate structures may play a role in gametocytogenesis and the maturation of the gametocyte. The multilaminate structures were either free in the cytoplasm of infected erythrocytes or present in association with the parasitophorous vacuole membrane surrounding immature gametocytes. These structures may originate from the erythrocyte plasma membrane and the parasitophorous vacuole membrane. Other notable findings in P. falciparum treated with cyclic AMP included the presence of loop-like membrane structures protruding from the plasma membrane of the parasite and termination of some plasma membranes of the parasite in dense granular structures.

Cell Membrane↗

Guanylate cyclase activity in human leukemic and normal lymphocytes. Enzyme inhibition and cytotoxicity of plant extracts.

Cyclic GMP is thought to be involved in lymphocytic cell proliferation and leukemogenesis. In general, the nucleotide is elevated in leukemic vs. normal lymphocytes and changes have been reported to occur during remission and relapse of this disease. Although the cA/cG ratios are higher for normal lymphocytes the basal guanylate cyclase (EC 4.6.1.2) activities do not correlate with altered cyclic GMP levels. The crude guanylate cyclases display classical Michaelis-Menten kinetics with Kms for Mn-GTP of 463 mumol/l and 20-90 mumol/l for normal and leukemic lymphocytes, respectively. An extract from the bitter melon (Momordica charantia) preferentially inhibits the soluble guanylate cyclase from leukemic lymphocytes. This inhibition correlates with its preferential cytotoxic effects for these same cells. Analyses of nine other cytotoxic plant extracts revealed that only an extract from the Lawson's cypress, Chamaecyparis lawsonianna, exhibits comparable cytotoxicity and guanylate cyclase inhibition levels.

Cyclic AMP↗

Purification and characterization of the entomocidal protoxin of Bacillus thuringiensis.

A procedure for purifying the insecticidal parasporal protoxin of Bacillus thuringiensis and a description of its biochemical and biophysical properties is provided. Mild alkali titration was necessary to generate a functional protoxin in a soluble form, and anion-exchange chromatography was used to remove contaminating cytoplasmic proteases that are nonspecifically bound to whole native parasporal crystals. Polyacrylamide gel electrophoresis, gel filtration chromatography, and meniscus depletion sedimentation equilibrium analysis revealed an apparent molecular weight for the protoxin of 1.34 x 10(5). The only NH2-terminal residue found was methionine. The soluble protoxin was 2.5 times more toxic to insect larvae than was the parasporal crystal. At alkaline pH the protoxin slowly converted to a low molecular weight toxin (apparent Mr = 6.8 x 10(4)). The molar specific toxicities of the protoxin and toxin were identical.

Amino Acid Sequence↗

Comparative biochemistry of entomocidal parasporal crystals of selected Bacillus thuringiensis strains.

Parasporal crystals of Bacillus thuringiensis subspp. kurstaki, tolworthi, alesti, berliner, and israelensis were compared by electron microscopy, polyacrylamide gel electrophoresis, amino acid analysis, tryptic peptide mapping, immunological analysis, and insecticidal activity. Spore coats also were compared by polyacrylamide gel electrophoresis. B. thuringiensis subsp. israelensis crystals were lethally toxic to mosquito larvae and nontoxic to tobacco hornworm larvae. Conversely, crystals from the other subspecies killed tobacco hornworm larvae but were ineffective against mosquitoes. Crystalline inclusion bodies of all subspecies contained a protoxic subunit that had an apparent molecular weight of approximately 1.34 X 10(5). However, polyacrylamide gel electrophoretic patterns of solubilized crystals revealed a small-molecular-weight component (apparent molecular weight, 26,000) in B. thuringiensis subsp. israelensis that was absent in the other subspecies. Also, differences were noted in amino acid composition and tryptic peptide fingerprints. Crystal proteins were found in spore coats of all subspecies. The results suggest that insecticidal specificity is due to unique polypeptide toxins.

Amino Acids↗

Interaction of thyroxine with lysozyme.

L-thyroxine and hen's egg-white lysozyme form an insoluble complex at neutral pH. Previous reports have suggested that the enzyme and hormone alos associate in solution. No evidence for the soluble interaction between thyroxine and lysozyme was obtained by using spectral, enzymatic, and immunological techniques.

Animals↗

Alterations in the distribution and proliferative responses of rhesus monkey peripheral blood and spleen cells during malaria (Plasmodium knowlesi) infection.

Rhesus monkeys are used frequently as animal models in malaria research, but few studies have evaluated lymphocyte functions in these animals after experimental infections with the primate malarial parasite Plasmodium knowlesi. In this study, the distribution and mitogen responses of mononuclear cells in the peripheral blood and spleens of 16 P. knowlesi-infected rhesus monkeys were followed. All animals included in the study developed acute infections and were bled out with parasitemias of more than 50%. With progression of the infection, alterations in the peripheral blood mononuclear cells were observed, including decreases in the percentage of T cells (measured by E rosette formation) and the total numbers of E and EAC rosette-forming cells per cubic millimeter. In addition, peripheral blood mononuclear cells displayed reduced responses to mitogen stimulation with phytohemagglutinin, concanavalin A, and pokeweed mitogen. Peripheral blood mononuclear cells from infected animals showed similar reductions in mitogen responses when cultured in media containing 15% autologous pre- or postinfection plasma. The mitogen responses of spleen cells did not appear to be affected, but a significant reduction in the proportion of splenic T cells was observed. These lymphocyte changes in P. knowlesi-infected rhesus monkeys are similar to those reported for mice with acute rodent malaria and for humans with chronic Plasmodium falciparum infections.

Animals↗