A ribonuclease (RNase) has been isolated from normal human pancreas obtained upon autopsy. About 5 mg of RNase is normally recovered per kilogram of pancreas, equivalent to ca. 70% of the total activity and a 700-fold purification from the initial acidified extract. The specific activity of the purified enzyme is identical with that of bovine pancreatic ribonuclease, and a single component is found in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, gel filtration, and reversed-phase high-pressure liquid chromatography. Aggregation of the protein is found upon ultracentrifugation under native and denaturing conditions, and several bands of equal specific activity are seen in polyacrylamide gel electrophoresis of the native protein. At least two components are glycoproteins. A molecular weight of 15 000 is estimated from sodium dodecyl sulfate gel electrophoresis, gel filtration, and amino acid and peptide analyses. The enzyme is related to bovine pancreatic RNase, but distinguishable by amino acid analysis, tryptic peptide maps, and low cross-reactivity of antibodies with the heterologous enzymes. The human enzyme is also inactivated by treatment with iodoacetic acid at pH 5.5 and is essentially identical with bovine RNase in its far-ultraviolet circular dichroism spectrum. The human RNase is like bovine pancreatic RNase catalytically; RNA is cleaved at pyrimidine residues, and activity against poly(cytidylic acid) is high.

Purification and characterization of human pancreatic ribonuclease.

Four major urine ribonuclease (RNase) activities, designated bands A-D, were identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and activity staining. Bands A, B, and C have alkaline pH optima and display molecular weights of 31 000, 23 000, and 20 000, respectively, upon sodium dodecyl sulfate (NaDodSO4) gel electrophoresis and weights of 44 000, 28 000, 22 000 upon gel filtration. Band D, with a pH optimum slightly below neutrality, has a molecular weight of 16 000 or 15 000, respectively, determined by the above methods. Band A, the most abundant activity in urine, is heterogeneous and resembles serum RNase 1 on electrophoresis and on phosphocellulose and Sephadex chromatography. Band B is similar to a minor, unnamed component of serum RNase activity while band C resembles serum RNase 3. Band D is similar to the leukocyte RNase-like activity of serum [Blank, A., & Dekker, C.A. (1981) Biochemistry (preceding paper in this issue)]. Band A is present in urine at a concentration high than that of RNase 1 in serum. In contrast, urine counterparts of serum RNases 2, 4, and 5 are not apparent upon either phosphocellulose chromatography [see also Yamanaka, M., Akagi, K., Murai, K., Hirao, N., Fujimi, S., & Omae, T. (1977) Clin. Chim. Acta 78, 191-201] or NaDodSO4 get electrophoresis; a urine counterpart of serum RNase 3 can be detected only by the more sensitive electrophoretic method. These results indicate that RNase 2-5 are processed differently by the kidney than RNase 1. After reconciliation of reported differences in their pH optima and molecular weights, five apparently diverse RNase preparations described in the literature can be related to band A activity and three preparations to band D. However, we are unable to confirm a previous report of a human urine enzyme indistinguishable from bovine pancreatic RNase A.

Multiple ribonucleases of human urine

Since Desialylated rebonuclease (RNase) from human urine has a highle alkaline isoelectric point (pI 9.5–10.5), an acidic buffer system is required for electrophoretic analysis of different urine specimens. In a common acid disc electrophoresis system in thin-layer polyacrylamide gel, the cathodal mobility of the RNase enzyme(s) was markedly reduced from that expected on the basis of the estimated pI. Dialysis urine against water had a very detrimental effect on the separation of the isozymes, and the addition of buffer to the dialyzed urine further exacerbated this effect. The urin RNase exhibited anomalous electrophoretic behavior in the form different urine specimens or from the same urine specimen adjusted to different concentrations. Treatment of urine with various cations had a significant, positive effect on the migration and resolution of the isozymes. This and other experimental evidence is consistent with the hypothesis that the anomalous behavior was due to the binding of anions from the buffer. A standardized protocol was developed for processing urine specimens, including treatment of urine with myoglobin, which abolished the artifactual variation observed previously and enabled reliable electrophoretic separation of the urine RNase isozymes. In addion, myoglobin treatment had a similar positive effect on the electrophoretic behavior of RNase enzymes from some other sources.

Anamalous electrophoresis behavior of rebonuclease from human urine and a novel treatment method for reliable analysis

The nutritional quality of artificial diets composed of pupal holotissues of Diatraea saccharalis (Fabr.) (Lep.: Crambidae) from three different age classes (1-2, 3-4 and 5-6 days old) were tested for rearing Trichogramma galloi Zucchi and T. pretiosum Riley (Hym.: Trichogrammatidae) in vitro. Pupal holotissues were added to egg yolk, bovine fetal serum, lactoalbumine hydrolysate and preservatives, and offered to the parasitoids into artificial eggs. The quality of these diets in supporting the development of both parasitoids was evaluated by assessing the acceptance and parasitism of the artificial host eggs, larval and pupal survival, size, parasitism capacity and longevity of the in vitro-reared females, and the presence of deformed adults. Diets composed of 5-6 d old pupal holotissues did not support the larval development of both parasitoids as well as reduced the acceptance and/or parasitism of artificial eggs by T. galloi and T. pretiosum. The factors affecting the nutritional quality of pupal holotissues collected from different developmental stages are discussed.

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Effect of the age of the pupal holotissue on the nutritional quality of artificial diets for Trichogramma spp. (Hymenoptera: Trichogrammatidae)

Lufenuron (chitin synthesis inhibitors) and Oryza sativa bran extract (waste product) exhibited insecticidal activity on females of Schistocerca gregaria when they were treated as one day old of the 5th nymphal instar with different concentrations of tested compounds. Some biological aspects, failure of ecdysis to adults, adult mortality, prolongation in the age of last nymphs and developmental events were investigated after treatment. Quantitative and qualitative determination of protein and DNA contents were investigated in normal and affected eggs which resulted from females Schistocerca   gregaria treated as one day old 5th nymphal instarwith LC50 ofLufenuron and Oryza sativa bran extract during embryogenesis. The total protein content of normal eggs at 0-day old, was 81.07 mg protein/gm eggs, after one day of oviposition  the total protein increase significantly (p<0.05). Then the protein content, during the early incubation period, to the 4th day post oviposition (pop) significantly (p<0.05) declined sharply to 38.74mg protein / gm eggs. While in the 7th, 11th &12th it was decreased significantly (p<0.05) during the late organogenesis stage. The treatment resulted a significant decrease in the total protein of rice bran extract affected eggs compared with the control at (1, 11& 12) days old eggs, and a significant increase at 0 and 4 days pop compared with control. Treatment with Lufenuron resulted a significant decrease of total protein in (0, 1 & 12) days old eggs compared with the control but there were no significant increase at 4, 7 and 11 old eggs. A total of (10 -12) protein bands were separated by electrophoresis during normal embryogenesis of Schistocerca   gregaria. Fraction protein of affectedeggs with LC50 of Lufenuron and rice bran extract resulted a separation of (10 – 14) and (11- 14) protein bands with molecular weights from 12 to 325 KDa. Data recorded significant differences between DNA content of normal and the affected Schistocerca gregaria eggs throughout embryogenesis (0-12 days) post-oviposition

https://eajbsf.journals.ekb.eg/article_41514_7fab03bd063c5fc0c60e44c56aefe57d.pdf

Toxicological and Biochemical Effects of Lufenuron and Rice Bran on Desert Locust, Schistocerca gregaria (Forskal) (Orthoptera: Acrididae)

Quantitative changes in DNA, RNA and protein during the entire developmental period—including embryogenesis, larval and pupal stages — in the house-fly were determined. The development of housefly from egg to adult takes around 8·5 days at 35 ± 1° C. During the course of embryonic development toal RNA and protein decreased. On the other hand, total DNA increased and registered peak value in 4 hr old eggs. The calorific content of the developing eggs showed a decrease from 5649 cal/g dry weight to 5361 cal/g dry weight indicating the utility of energy reserves during embryonic development. Matured larvae registered high calorific content (6260 cal/g dry weight).

A Srinivasan

Papers

Biochemical characterisation of the development ofMusca domestica