Anti-CD38 prevents the development of the adaptive response induced by X-rays in human lymphocytes

Irradiation of human lymphocytes (1 cGy X-rays, 37 degrees C) evoked an approximately 30% decrease in the frequency of micronuclei upon subsequent X-irradiation (1.5 Gy). The response was reflected in a lower micronucleus frequency but not in the DNA repair rate measured by the comet assay directly after the challenge dose. Treatment of lymphocytes with anti-CD38 antibody 1 h before irradiation with the adaptive dose prevented the development of the adaptive response measured as micronuclei frequency, but adaptation was not reflected in a lower rate of DNA repair, measured by the alkaline version of the 'comet' assay. In lymphocytes that were anti-CD38-treated and irradiated and or irradiated with the adaptive dose the rate of DNA repair was not changed. However, the mean DNA damage level in adapted anti-CD38-treated lymphocytes was significantly lower than that in the control lymphocytes at all time points. We conclude that ligation of CD38 by antibody initiates signalling that prevents the development of the adaptive response induced by X-rays. Lower chromosome damage revealed by the cytokinesis block-micronucleus test in the adapted lymphocytes is unrelated to DNA repair rate.     

A major isoform of the maize plasma membrane H(+)-ATPase: characterization and induction by auxin in coleoptiles

The plasma membrane (PM) H(+)-ATPase has been proposed to play important transport and regulatory roles in plant physiology, including its participation in auxin-induced acidification in coleoptile segments. This enzyme is encoded by a family of genes differing in tissue distribution, regulation, and expression level. A major expressed isoform of the maize PM H(+)-ATPase (MHA2) has been characterized. RNA gel blot analysis indicated that MHA2 is expressed in all maize organs, with highest levels being in the roots. In situ hybridization of sections from maize seedlings indicated enriched expression of MHA2 in stomatal guard cells, phloem cells, and root epidermal cells. MHA2 mRNA was induced threefold when nonvascular parts of the coleoptile segments were treated with auxin. This induction correlates with auxin-triggered proton extrusion by the same part of the segments. The PM H(+)-ATPase in the vascular bundies does not contribute significantly to auxin-induced acidification, is not regulated by auxin, and masks the auxin effect in extracts of whole coleoptile segments. We conclude that auxin-induced acidification in coleoptile segments most often occurs in the nonvascular tissue and is mediated, at least in part, by increased levels of MHA2.     

The catalytic core of RNase P

A deletion mutant of the catalytic RNA component of Escherichia coli RNase P missing residues 87-241 retains the ability to interact with the protein component to form a functional catalyst. The deletion of this phylogenetically conserved region significantly increases the Km, indicating that the deleted structures may be important for binding to the precursor tRNA substrate but not for the cleavage reaction. Under some reaction conditions, this RNase P deletion mutant can become a relatively non-specific nuclease, indicating that this RNA's catalytic center may be more exposed. The catalytic core of the RNase P is formed by less than one third of the 377 residues of the RNase P RNA.     

A specific antidote for the treatment of digitalis poisoning in uremic patients

Two chronic haemodialyzed patients with digitalis intoxication are reported. One of them took digoxin 0.25 mg three times daily for an unknown period and the other took digitoxin 0.1 mg twice daily for two weeks. The symptoms of intoxication were mainly concealed by uremic syndrome. The diagnosis was established by noticed sinus bradycardia, first- and second-degree atrioventricular block in ECG and the determination of sera levels of glycosides (serum digoxin concentration was 7.36 ng/ml, serum digitoxin concentration was 46.5 ng/ml) in both cases. Considering the probable long elimination period of digitalis and the potentially life-threatening situation the patients were given digoxin-specific antibody (Fab) fragments with potassium replacement therapy. The symptoms disappeared within a few hours after therapy, side effects and rebound toxicity did not develop. In connection with these cases the aim of this report is to publish a method which can reverse the life-threatening digitalis intoxication in patients suffering from renal failure as well. As to the above method, the authors have not found any similar case reports in the Hungarian medical literature.     

Social constructivism and capacity building for environmental governance

The concept of capacity building for environmental protection and management refers to the process by which the abilities of societies to identify and solve environmental problems are developed. This work assumes a social constructivist perspective to the process of capacity building for environmental governance and thus depicts it as critically dependent on broader processes of political and socioeconomic modernization. In an attempt to depict such a process, a heuristic device is put forward as a model of capacity analysis. A theoretical discourse and new institutionalist perspectives are used to test such a model on a theoretical plane. Some preliminary conclusions are drawn from this theoretical exercise.     

Local staging with CT of tumors of the upper urothelium

OBJECTIVE: To evaluate the ability of computerized tomography (CT) to stage transitional cell carcinoma of the upper urinary tract. METHODS: 29 transitional cell carcinoma of the upper urinary tract submitted to nephroureterectomy were retrospectively evaluated. All 29 tumors had preoperative CT scans performed to stage the lesion. The pathological staging was compared to that of CT. RESULTS: 10 of the 29 tumors had CT evidence of tumor extension and 19 had localized noninvasive tumor on CT. Of the 10 patients with CT findings of tumor extension, 2 (20%) had superficial tumors and 8 (80%) had tumors that invaded into the adventitial fat, renal parenchyma or perirenal fat (pT3, pT4). Of the 19 patients with localized noninvasive tumor on CT, 13 (68%) had superficial tumors and 6 (32%) had pT3 or pT4 tumors. CT sensitivity for tumor invasion was 57% with a specificity of 87.5%. CONCLUSIONS: Our analysis shows that CT is of limited value in staging these tumors. When CT demonstrates direct tumor extension through the renal pelvic or ureteral wall, it is a sensitive indicator of high-stage tumor. However, the results obtained in low stage tumors must be viewed with caution.     

A column centrifugation method for the reconstitution in liposomes of the mitochondrial F0F1 ATP synthase/ATPase

A method for reconstitution of membrane proteins into unilamellar liposomes is described. The model enzyme was the F0F1 ATP synthase from mitochondria when in complex or free from its inhibitor protein. The enzymes were first solubilized with either of two detergents, i.e., n-dodecyl-beta-D maltoside or lauryldimethylamine oxide. After solubilization, the enzymes were passed through a column of Sepharose-AH using an ADP/sodium cholate selective elution buffer. The enzymes recovered from the column were subsequently passed through a centrifuge column of Sephadex G-50 fine. The eluate contained liposomes in which the F0F1 complex (with and without inhibitor protein) had been reconstituted. The reconstituted enzymes were capable of hydrolyzing ATP with formation of electrochemical H+ gradients. They also catalyzed the ATP-Pi exchange reactions. Thus the F0F1 complex which is formed by 18 subunits can be rapidly reconstituted into liposomes in a fully functional state. Moreover the data show that the interactions between the enzyme and its inhibitor protein are not perturbed in the reconstitution procedure.     

Fracture of glass in tensile and bending tests

To fully characterize the mechanical behavior of ceramics, it is useful to study their response to both tensile and bending tests. In this investigation, a quantitative comparison is made between tensile and bending results from (circularly) cylindrical glass rods of borosilicate glass and of fused silica. These experimental results show that in each material, the Weibull exponentm is approximately the same for both types of test. The fractured samples clearly showed that all failures initiated at surface-located flaws. The stress level at which the probability of fracture in tension equals 50 Pct was determined; the maximum tensile stress for 50 Pct probability of fracture in bending was also determined. The ratio of these two stresses was predicted previously from simple, theoretical considerations based upon the applicability of the Weibull function and on the distribution of defects in the specimens. This theoretical result is inconsistent with the experimental observations in view of the fractography. Therefore, it is concluded that failure in the materials tested is not satisfactorily described by Weibull statistics.     

Properties of permease dimer, a fusion protein containing two lactose permease molecules from Escherichia coli

An engineered fusion protein containing two tandem lactose permease molecules (permease dimer) exhibits high transport activity and is used to test the phenomenon of negative dominance. Introduction of the mutation Glu-325-->Cys into either the first or the second half of the dimer results in a 50% decrease in activity, whereas introduction of the mutation into both halves of the dimer abolishes transport. Lactose transport by permease dimer is completely inactivated by N-ethylmaleimide; however, 40-45% activity is retained after N-ethylmaleimide treatment when either the first or the second half of the dimer is replaced with a mutant devoid of cysteine residues. The observations demonstrate that both halves of the fusion protein are equally active and suggest that each half may function independently. To test the possibility that oligomerization between dimers might account for the findings, a permease dimer was constructed that contains two different deletion mutants that complement functionally when expressed as untethered molecules. Because this construct does not catalyze lactose transport to any extent whatsoever, it is unlikely that the two halves of the dimer interact or that there is an oligomeric interaction between dimers. The approach is consistent with the contention that the functional unit of lactose permease is a monomer.