Ribosome-inactivating proteins depurinate poly(ADP-ribosyl)ated poly(ADP-ribose) polymerase and have transforming activity for 3T3 fibroblasts

It has been known that ribosome-inactivating proteins (RIPs) from plants damage ribosomes by removing adenine from a precise position of rRNA. Subsequently it was observed that all tested RIPs depurinate DNA, and some of them also non-ribosomal RNAs and poly(A), hence the denomination of adenine polynucleotide glycosylases was proposed. We report now that ricin, saporin-L2, saporin-S6, gelonin and momordin depurinate also poly(ADP-ribosyl)ated poly(ADP-ribose) polymerase (auto modified enzyme), an enzyme involved in DNA repair. We observed also that all RIPs but gelonin induce transformation of fibroblasts, possibly as a consequence of damage to DNA and of the altered DNA repair system.     

An immunotoxin containing a rat IgM monoclonal antibody (Campath 1) and saporin 6: effect on T lymphocytes and hemopoietic cells

Summary The elimination of the cells responsible for graft-versus-host disease in allogeneic bone marrow transplantation has been attempted with a variety of methods, including the use of the ribosome-inactivating toxin ricin bound to monoclonal antibodies acting as carriers. However the high nonspecific toxicity of these immunotoxins containing the whole toxin greatly limited clinical application. Toxicity can be reduced using the A-chain of ricin or other ribosome-inactivating proteins (RIPs) which are devoid of a B-chain with lectin properties. We used saporin 6 purified from Saponaria officinalis seeds, which was conjugated with the rat IgM monoclonal antibody Campath 1 specific for mature T and B lymphocytes as well as for monocytes. The immunotoxin retained both RIP and antibody activity, inhibiting protein synthesis both in a cellfree system and in cells bearing the Campath 1 antigen; it also abolished methyl ³H-thymidine uptake in phytohemagglutinin-stimulated T lymphocytes. Myeloid progenitors were largely spared as shown by myeloid stem cell (CFUGM) growth which was scarcely affected. Toxicity of the immunotoxin to cell lines not expressing the antigen recognized by Campath 1 monoclonal antibody was not greater than the toxicity due to free saporin 6, while the immunotoxin was more toxic to mice than free saporin.Work supported by grants of Regione Emilia Romagna, delibera n. 1970, 13/5/86, by the Italian National Research Council, Roma, Finalized Project Oncologia, contracts n. 86.00589.44 and n. 86.00603.44 and by the Pallotti's Legacy for Cancer ResearchAngelo Dinota is supported by a grant from Associazione Italiana per la Ricerca sul Cancro (AIRC), Milan, Italy.     

The ecology and morphology of Australia's desert turtle (Emydura macquarii emmotti)

Cooper Creek is one of Australia's largest unregulated river systems and one of the world's most variable large river systems. It is a dynamic environment that oscillates between booms and busts; yet, many species thrive in it. One of these species, the Cooper Creek turtle (Emydura macquarii emmotti) has received little attention, despite being one of Australia's largest freshwater turtles and living further inland than any other Australian turtle. We conducted surveys for E. m. emmotti in 2001–2004, 2019, and 2022, focussing predominantly on the Waterloo waterhole. Waterloo had a large population of E. m. emmotti (508 estimated individuals; 95% CI = 447–596) with an estimated density of 64.8 turtles/ha (95% CI = 57.0–76.2) and estimate biomass of 74.4 kg/ha (95% CI = 57.6–100.3 kg/ha). Juveniles were highly abundant in all years, representing up to 63.6% of captured individuals. It was slightly (but not significantly) male-biased in 2001–2004 and significantly female-biased in 2019. All sizes and sexes used the floodplain during a flooding event in 2022, but more males than females were captured on the floodplain, and there was evidence of male-biased dispersal across the years. Compared to Murray River turtles (Emydura macquarii macquarii), E. m. emmotti exhibited megacephaly across all ages and sexes, with particularly pronounced megacephaly in adult females. Algae were present on many individuals (including on the skin and plastron) but was relatively more abundant on juveniles. Leeches were not detected on any of the 66 turtles that were examined for them. The following injuries/malformations were noted: missing or injured limbs (3.2%), missing or injured eyes (1.3%), damaged shells (8.0%), scute/shell anomalies and malformations (10.6%), and marginal scute seams extending into the costals (67.4% of adults, 1.2% of juveniles). This paper presents some of the first work on this unusual turtle and makes recommendations for future research.     

Inhibition of protein synthesis by ricin: experiments with rat liver mitochondria and nuclei and with ribosomes from Escherichia coli (Short Communication)

1. Ricin, a toxic protein from the seeds of Ricinus communis which inhibits poly(U)-directed polyphenylalanine synthesis by rat liver ribosomes (Montanaro et al., 1973), does not affect protein synthesis by isolated rat liver mitochondria. 2. The toxin is ineffective also on poly(U)-directed polyphenylalanine synthesis in reconstituted systems with ribosomes isolated from rat liver mitochondria or from Escherichia coli. 3. Ricin inhibits protein synthesis by isolated rat liver nuclei, but at concentrations much higher than those affecting rat liver ribosomes.     

Histidyl-proline diketopiperazine: its biological role as a regulatory peptide

Summary Histidyl-proline diketopiperazine [cyclo(His-Pro)] is a metabolic of thyrotropin releasing hormone (TRH). This review summarizes the literature concerning cyclo (His-Pro) and, in addition, some studies dealing with TRH and other peptides that are considered of interest. The enzymes concerned with the metabolism of TRH are discussed. Distribution studies of peptides by immunological methods show that, while TRH is concentrated in synaptosomes, cyclo (His-Pro) is not, suggesting that cyclo (His-Pro) is not a classical neurotransmitter. Rat brain contains approximately three times as much cyclo (His-Pro) as TRH, mainly localized in the pituitary and hypothalamus. While the TRH is found in a free form, the cyclo (His-Pro) is bound to a carrier of molecular weight approximately 70 000. While specific membrane receptors for TRH have been detected in pituitary cells, no such receptors for cyclo (His-Pro) have yet been found in brain or pituitary; however, there is a specific binding of cyclo (His-Pro) to adrenal cortex membranes, Both TRH and cyclo (His-Pro) have effects in the central nervous system or pituitary. These include effects on prolactin release, thermoregulation, CNS depression, stereotypic behavior and cyclic nucleotide levels. Possible mechanisms and interrelations of these effects are discussed.     

Inhibition by ricin of protein synthesis in vitro. Ribosomes as the target of the toxin

1. Ricin (a toxic protein from the seeds of Ricinus communis) is a powerful inhibitor of the poly(U)-directed incorporation of phenylalanine into polypeptides catalysed by isolated rat liver ribosomes and elongation factors 1 and 2 (EF 1 and EF 2). The inhibition can be largely overcome by increasing the concentration of ribosomes. 2. The toxin does not affect the binding of phenylalanyl-tRNA to ribosomes catalysed by EF 1, nor does it inhibit the puromycin reaction used as a test for peptide-bond formation catalysed by ribosomes. 3. Ricin inhibits the ribosome-linked GTP hydrolysis catalysed by EF 2. 4. Ribosomes treated with ricin and washed through sucrose gradients containing 0.6m-NH(4)Cl are functionally inactive in those assay systems that are sensitive to the presence of added toxin. 5. It is suggested that ricin brings about an irreversible modification of ribosomes which impairs their ability to interact with EF 2. Since ricin inhibits at a molar concentration much lower than that of ribosomes it probably acts catalytically. No added cofactor is necessary for the inhibitory action of the toxin.     

Continuity or discontinuity of the life-style in central Italy during the Roman Imperial Age-Early Middle Ages transition: diet, health, and behavior

Dento-alveolar pathologies and alterations (dental wear, caries, abscesses, ante mortem tooth loss (AMTL), calculus, hypoplastic defects, and chipping) and skeletal markers of health (cribra orbitalia and periostitis) were analyzed in two skeletal samples from the necropolises of Quadrella (I-IV c. AD) and Vicenne-Campochiaro (VII c. AD) in the Molise region of central Italy. The aim was to determine if the Roman Imperial Age-Early Middle Ages transition characterized by political, socioeconomic, and cultural transformations affected the biology of these populations, particularly their alimentation and health status. The frequencies of caries and AMTL, similar in the two samples, suggest a high consumption of carbohydrates. The higher levels of heavy wear, calculus, and interproximal chipping in the Vicenne population indicate a greater use of fibrous foods (both meat and others), in line with the dietary model of Germanic peoples. Health conditions do not appear to have been good in either period, as shown by the high frequencies of linear hypoplasia and the presence of cribra orbitalia and periostitis. The diet of the individuals buried with horses of the Vicenne population did not differ from that of the rest of the population, whereas there were evident differences in the use of the teeth for nonmasticatory activities among these individuals. Therefore, from the point of view of alimentation and health status, the profound socioeconomic and cultural transformations during the Late Antiquity-Early Middle Ages transition do not seem to have been translated into a true discontinuity of the two Molisan populations.