The exotoxin ofBacillus thuringiensis: A new C-mitotic agent

Summary Thuringiensin A, an exotoxin fromBacillus thuringiensis, a constituent of the microbial insecticide thuricide has been found to inhibit mitotic spindle, condense and scatter chromosomes. It may therefore be a promizing tool in future cell biological studies.We thank Prof.H. Sharatchandra for facilities. CBSR is a National Associate of the University Grants Commission, New Delhi.     

Antagonism of terramycin on action ofBacillus thuringiensis ‘exotoxin’ inDrosophila melanogaster

Zusammenfassung Subletale Konzentrationen von Terramycin hemmen inDrosophila teilweise die toxische Wirkung des hitzestabilen Exotoxins vonBacillus thuringiensis, wobei die Steilheit der Dosis-Mortalitätskurven nicht verändert wird. Es wird die Hypothese begründet, dass es sich um eine kompetitive Hemmung des Exotoxins durch das Antibioticum handeln könnte.     

On the chemical nature of the heat-stable exotoxin ofBacillus thuringiensis

Zusammenfassung Das hitzestabile Exotoxin vonBacillusthuringiensis — auf Insekten spezifisch toxisch wirkend — wurde aus dem Kulturmedium angereichert und stark gereinigt. Sämtliche toxischen Fraktionen zeigen ein Absorptionsmaximum bei 258–260 nm, ein Absorptionsminimum bei 232–234 nm und eine positive Aldopentosereaktion. Bei der toxischen Substanz scheint es sich um ein Nucleosid oder Nucleotid des Adenins oder Uracils zu handeln.     

The toxic action ofbacillus thuringiensis ‘Exotoxin’ onDrosophila reared in yeast-containing and yeast-free media

Zusammenfassung Die toxische Wirkung des sogenannten «Exotoxins» vonBacillus thuringiensis aufDrosophila wurde geprüft: (A) in Mais-Trockenhefe-Agar-Medium, (B) synthetischem Medium C nachSang ² und (C) synthetischem Medium plus 2% Trockenhefe. In hefefreiem Medium ist die LC₅₀ bedeutend niedriger und die Probitkurve viel steiler als in hefehaltigen Medien. Hefe reduziert also die toxische Wirkung des «Exotoxins» beträchtlich.

---

---

Supported by Fellowship No. 1560 of the National Research Counsel of Canada.     

Nachweis von Nahrungsmittel-Antikörpern mittels der Hämagglutination

Summary By means of ahaemo-agglutination test using erythrocytes sensitised with tannic acid, we were able to demonstrate the presence of cow's milk antibodies in humans. The use of the haemo-agglutination test enables us now to give exact information on the titer value of the antibody in human serum. Previously this was limitately possible using the method of antibody demonstration by means of the complement-compound reaction.We have well-established reasons to assume that other antibodies caused by various antigens, can be determined by means of the haemo-agglutination test.     

Presence of anti-Trypanosoma cruzi antibodies in the sera of mice with experimental autoimmune myocarditis

Summary The existence of antigens shared in common byT. cruzi and heart muscle cells is suggested by the presence of antibodies binding to the parasite surface in the serum of mice with autoimmune myocarditis induced by immunization with syngeneic heart antigens.     

Alteration of capsular type of encapsulated strains ofStaphylococcus aureus during freeze-drying and storage

Summary Remarkable alteration was shown in capsular type antigen production in encapsulated strains ofStaphylococcus aureus stored by lyophilization for 10 years. This alteration was further elucidated by antibody production in rabbits immunized with the altered strain and by absorbing the antibodies with representative capsular type strains.     

Oral and parenteral toxicity ofBacillus thuringiensis ‘exotoxin’, and its inactivation in larvae ofGalleria mellonella

Zusammenfassung Es wird gezeigt, dass Larven der grossen Wachsmotte durch oral appliziertes «Exotoxin» vonBacillus thuringiensis vergiftet werden, die LD₅₀ aber ca. 250mal grösser ist als bei parenteraler Applikation. Oral verabreichtes «Exotoxin» wird zu einem grossen Teil im Raupendarm inaktiviert. Es konnte eine mathematische Beziehung zwischen der Inkubationszeit und der Inaktivierung des «Exotoxins» gefunden werden, die für einen enzymatischen Abbau des «Exotoxins» spricht.     

Anti-bovineβ-lactoglobulin antibodies react with a human lactoferrin fragment and bovineβ-lactoglobulin present in human milk

Human milk samples react against anti-bovine-lactoglobulin rabbit antibodies, as measured by a competitive radioimmunoassay. Immunoreactivity was positive even in milk from mothers consuming a diet free of cow's milk. An increase with a diet rich in cow's milk proteins was detected by immunoelectrophoresis. The human milk fraction cross-reacting with anti-bovine-lactoglobulin antibodies corresponds to the 20 kDa fragment from the N-terminal end of human lactoferrin. Three regions of this fragment exhibit sequence homology with a sequence contained in cow's-lactoglobulin (between residues 124 and 141).     

The effect of continuously cooling the hypothalamic preoptic area on antibody titre in the rat

Summary Continuous cooling of the hypothalamic preoptic area for 5 days, as well as sublethal infection withSalmonella enteritidis, increased the titre of antibodies against sheep erythrocytes, suggesting that the febrile response stimulates the humoral immune response.We thank Dr W. Mannheim from the Institute of Hygiene for kindly supplying the cultures ofS. enteritidis. This work was supported by the Schwerpunktprogramm Temperaturregulation und-adaptation of the Deutsche Forschungsgemeinschaft.     

Production of antibodies to amanitins as the basis for their radioimmunoassay

Summary The production of antibodies against amanitins is described. By means of these antibodies, a radioimmunoassay was developed which allows detection of as little as 0.5 ng of amanitins in 1 ml of serum. By this method, the clearance of α-amanitin from the blood of poisoned mice was measured. Acknowledgments. We thank ProfessorT. Wieland and Dr.H. Faulstich (Heidelberg) for their generous gift of β-amanitin and [³H]O-methyl-demethyl-γ-amanitin. This work was supported by grants from C.N.R., Rome.     

Antibody-induced formation of caps inToxoplasma gondii

Summary Trophozoites ofToxoplasma gondii from mouse peritoneal exudate move their surface membrane antigens towards one pole of the cell when incubated with antibodies. The phenomenon may be induced in up to 50% of incubated parasites. It is prvented by some metabolic inhibitors and low temperatures (0–4°C). These properties do not change in parasites subpassaged after repeated incubation with antibodies.We wish to thank Dr. J.lusarczyk of the National Institute of Hygiene for his help in preparation the photomicropgraphs.     

Monoclonal antibodies to L-asparaginase

Summary Five hybridomas secreting monoclonal antibody toE. coli L-asparaginase were isolated. These monoclonal antibodies were classified into 3 different subclasses; Ig G₁ (1 clone), Ig G₂ (2 clones) and Ig G₃ (2 clones). One of them possessed anti-L-asparaginase neutralizing activity. Four antibodies examined demonstrated a linear Langmuir binding plot and binding affinities, with equilibrium dissociation constant (K_d) ranging between 2.5×10^–9M and 6.3×10^–10 M. The monoclonal antibodies should be useful probes for investigation of the enzyme activity.     

Production and characterization of antibodies cross-reactive with major aflatoxins

Antibodies cross-reactive with 4 major aflatoxins were demonstrated three weeks after immunization of rabbits with an immunogen which was prepared by conjugating aflatoxin B₃ to bovine serum albumin. Aflatoxin B₃ was first converted to its hemisuccinate before conjugation to the protein. Tritiated aflatoxin B₁ (AFB₁) was used as the marker ligand both for antibody titer determination as well as for analysis of antibody specificity. Competitive RIA revealed that the antibodies have good cross-reactivity with aflatoxins B₁, B₂, G₁, and G₂ when tritiated AFB₁ was used as the marker ligand. The concentrations causing 50% inhibition of binding of³H-AFB₁ to the antibodies by unlabeled aflatoxins B₁, B₂, G₁, G₂ and B₃ were found to be 0.25, 3.34, 0.32, 4.0 and 0.53 ng/assay, respectively. The antibodies could be used for simultaneous analysis of aflatoxins B₁ and G₁, two of the most important toxic metabolites produced byAspergillus flavus andA. parasiticus.     

Evidence for a role of IFN γ in control ofListeria monocytogenes in T cell deficient mice

Summary The role of interferon (IFN) in controlling chronic infections ofListeria monocytogenes (Listeria) was studied in athymic C57BL/6nu/nu mice, and by treating thymectomized C57BL/6 +/+ mice with monoclonal rat CD4 and CD8-specific monoclonal antibodies (Mab). Mice treated with a combination of the two T cell subset antibodies were similar to athymic, nude mice in being able to contol Listeria infection, keeping the titers below 3–5 log₁₀ bacteria per organ, but they could not eliminate them completely. Treatment with antibodies to IFN of nude or CD4⁺ + CD8⁺-T cell-depleted mice suffering from chronic Listeria infection caused a marked increase of Listeria titers, in liver and spleen. This result implies a role of IFN in maintaining anti-Listeria resistance in mice lacking mature T cells.     

Gewinnung präzipitierender Insulin-Antikörper von der Ziege,Caper domesticus

Summary Antiserum was produced against beef insulin in male goats (goat anti-insulin-serum). It contains precipitating insulin antibodies; they were identified by immunelectrophoresis as a fraction of-globulin.     

Insulin activates Gαil,2 protein in rat hepatoma (HTC) cell membranes

Insulin action is initiated by binding to its cognate receptor, which then triggers multiple cellular responses by activating different signaling pathways. There is evidence that insulin receptor signaling may involve G protein activation in different target cells. We have studied the activation of G proteins in rat hepatoma (HTC) cells. We found that insulin stimulated binding of guanosine 5′-O-(3-thiotriphosphate) (GTP-γ-³⁵S) to plasma membrane proteins of HTC cells, in a dose-dependent manner. This effect was completely blocked by pertussis toxin treatment of the membranes, suggesting the involvement of G proteins of the Gα _i/Gα _o family. The expression of these Gα proteins was checked by Western blotting. Next, we used blocking antibodies to sort out the specific Gα protein activated by insulin stimulation. Anti-Gα _il,2 antibodies completely prevented insulin-stimulated GTP binding, whereas anti-Gα _o,i3 did not modify this effect of insulin on GTP binding. Moreover, we found physical association of the insulin receptor with Gα _i1,2 by copurification studies. These results further support the involvement of a pertussis toxin-sensitive G protein in insulin receptor signaling and provides some evidence of specific association and activation of Gα _i1,2 protein by insulin. These findings suggest that Gα _i1,2 proteins might be involved in insulin action. Received 23 September 1998; received after revision 23 November 1998; accepted 25 November 1998     

Suppression and enhancement of humoral immune response toToxoplasma gondii by passive antibody

Summary Anti-toxoplasma antibodies administered passively to mice may lead to suppression or enhancement (reported for the first time with Protozoan parasites) of subsequent antibody response when these animals are later infected withToxoplasma gondii. The outcome is dependent on infecting strain of Toxoplasma and the antigen-antibody ratio.     

Über die Beziehungen zwischen Albumin-γ-Globulin-Quotienten und der immunologischen Reaktionsfähigkeit

Summary 169 cats were immunized with heat-killed antigen ofBrucella abortus. No correlation between the albumin--globulin-quotient and the level of agglutinating antibodies could be demonstrated.     

Using a recombinant bispecific antibody to block Na<Superscript>+</Superscript>-channel toxins protects against experimental scorpion envenoming

In recent years, several molecular engineering methods of designing bispecific antibodies in various formats have been developed. Tandem-scFvs comprising two scFvs fused together via a peptide are 55-kDa molecules, and are one of the most promising and most straightforward approaches to bispecific antibody production. We report an attempt to design more effective antivenoms to the Androctonus australis scorpion using murine scFvs as building blocks to create a unique bispecific molecule that neutralizes the potent neurotoxins AahI and AahII. The tandem-scFv was produced in recombinant bacteria, purified by immobilized metal ion affinity chromatography, and analyzed by polyacrylamide gel electrophoresis, Western blot, gel filtration, mass spectrometry, and direct and competitive radioimmunoassay. In vivo, it neutralized the binding of the AahI and AahII toxins to their receptor, and protected mice against experimental envenomation. The findings reported here highlight the potential of recombinant antibody fragments for protecting against scorpion venom toxicity. Received 8 September 2006; received after revision 10 November 2006; accepted 27 November 2006