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Biomedical subjects

H Hardt

Publications and source records attributed to H Hardt.

At least 19 recordsLinked to original sources

Altered phosphoglycerate kinase from old rat muscle shows no change in primary structure.

Phosphoglycerate kinase (ATP:3-phospho-D-glycerate 1-phosphotransferase, EC 2.7.2.3) from young and old rat muscle was purified to homogeneity. After ascertaining that each preparation of the enzyme obtained from the latter indeed possessed altered properties, matched pairs of young and old enzymes were subjected to amino acid analysis and peptide mapping by HPLC. Following S-carboxymethylation, the respective young and old enzymes were digested with each of the following three proteinases: trypsin, chymotrypsin and S. aureus V8 proteinase. The corresponding peptides were resolved by reverse-phase HPLC. The peptide patterns obtained from both enzyme forms were identical. Even when the peptides obtained from digestion of phosphoglycerate kinase with S. aureus V8 proteinase were further digested with trypsin, no differences were observed. Comparative amino acid analyses also showed no differences. These results provide direct evidence that there are no changes in the sequence of altered rat muscle phosphoglycerate kinase and support the hypothesis that the differences in properties between the young and old forms of the enzyme result from a conformational modification.

Aging↗

The involvement of a high potential acceptor in the acid-base induced reduction of the acceptor Q in chloroplasts.

Study of the acid-base induced fluorescence transient, the so-called 'reverse electron flow', in chloroplasts revealed the following new properties: (1) Experiments in which the acid-base transition was performed in the absence of the measuring beam showed that the high fluorescence state induced by a pH-jump was attained and decayed even in complete darkness. These results indicated that a non-photochemical electron transfer was affected by the pH transition. A pH-induced reverse electron transfer from some secondary electron acceptor to Q probably occurred during that process. (2) This conclusion was supported by the effect of the Photosystem I electron acceptor methyl viologen. Methyl viologen accelerated the decay phase of the transient showing that this phase was controlled by the rate of electron flow to Photosystem I, but this acceptor did not diminish the size of the transient's initial rise, probably because this rise reflected a pH effect on a non-photochemical step located between Q and P-700. (3) The size of the fluorescence transient was dependent upon the reduction state of both parts of the secondary pool of electron acceptors, A2 and A1. (4) Redox potential measurements using ferricyanide-ferrocyanide mixtures showed that the size of the transient was directly dependent on a midpoint potential of +385 mV at pH 6.9 and with n = 1. This suggested the involvement of a high potential secondary electron acceptor in the acid-base induced reduction of Q.

Chloroplasts↗

Comparison of photosynthetic activities of spinach chloroplasts with those of corn mesophyll and corn bundle sheath tissue.

Bundle sheath and mesophyll chloroplasts from Zea mays showed comparable rates of O(2) evolution, which amounted to about half of the rate observed in spinach (Spinacia oleracea) chloroplasts.Ratios of 4.5, 4.6, and 6.2 Mn(2+) atoms per 400 chlorophylls were observed in mesophyll, bundle sheath, and spinach chloroplasts, respectively. These ratios roughly correspond to the observed O(2) evolution rates.Rates of electron transport from water to methylviologen (photosystem I and II) in both types of corn chloroplasts were about one-third that in spinach. Compared to spinach, transport rates from reduced diaminodurene to methylviologen (photosystem I) were about one-third and greater than one-half in mesophyll and bundle sheath material, respectively.In both types of corn chloroplasts, electron flow from photosystem II to P700 was abnormal. This observation, together with the low rates of all activities, suggests that damage occurred during isolation. Such damage may limit the quantitative significance of observations made with these materials (including the following data).Measurements of flash yields of O(2) evolution or O(2) uptake showed that the size of the photosynthetic unit was the same in photosystems I and II and in all three types of chloroplasts (about 400 chlorophylls per equivalent).Similarity of the photochemical cross-section of the two photosystems in the three preparations was also found in optical experiments: that is the half-times of the fluorescence rise in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) (photosystem II) and of the photooxidation of P700 (photosystem I).The ratio of P700 to chlorophyll appeared to be about 2-fold higher in bundle sheath chloroplasts than in the other materials (1/200 versus 1/400).

Journal Article↗

[Gamma scan findings in various bone processes in the jaw region].

Bone scintigraphy is a new means to early diagnosis of primary and secondary bone tumors, before significant radiological changes occur. The method is used with the osteotrope radioisotope 99mTc. Scintigraphy is most important in the determination of extent and localization of malignancies, relative to the indication for surgery, the tactics of surgery and for the diagnosis of recidives. In osteomyelitis, it facilitates early discovery and judgement of a florid or chronic process.

Bone Diseases↗

Plastocyanin as the possible site of photosynthetic electron transport inhibition by glutaraldehyde.

Treatment of spinach chloroplasts with glutaraldehyde causes an inhibition in the electron transport chain between the two photosystems. Measurements of O(2) flash yields, pH exchange, and fluorescence induction show that the O(2) evolving apparatus, photosystem II and its electron acceptor pool are not affected. The behavior of P700 indicates that its reduction but not its oxidation, is severely inhibited. Cytochrome f is still reducible by photosystem II but also slowly oxidizable by photosystem I. The sensitivity of isolated plastocyanin to glutaraldehyde further supports the conclusion that glutaraldehyde inhibits at the plastocyanin level and thereby induces a break between P700 and cytochrome f.

Journal Article↗

Stabilization by glutaraldehyde of high-rate electron transport in isolated chloroplasts.

Treatment of isolated chloroplasts with glutaraldehyde affects their ability to photoreduce artificial electron acceptors. The remaining rate of O2 evolution approaches zero with methyl viologen, is low with ferricyanide, but nearly normal with lipophilic Photosystem II acceptors, like oxidized p-phenylenediamine and oxidized diaminodurene. Since Photosystem I donor reactions are also affected, a specific site of inhibition of electron transport to Photosystem I is indicated. At the same time, glutaraldehyde prolongs the longevity of the chloroplasts stored in dark. In control samples the half-life of Photosystem II activity varied between 5 days at 4degreesC and 1 day at 25degreesC. Glutaraldehyde treatment increased these half times approx. 3-fold. The glutaraldehyde doses required to induce inhibition and stabilization were very similar.

Aldehydes↗

Circular polarization of fluorescence of chlorophyll in solution and in native structures.

Chlorophyll dimers in solution, subchlorplast particles and chloroplasts were investigated by their circular dichroism and circular polarization of their fluorescence, which reflect their optical rotatory power in the ground state and electronically excited state, respectively. The chlorophyll dimers in fluid solution lose their optical activity upon electronic excitation, reflecting a marked concomitant change in the structure of the dimers. This change is arrested in a solution of very high viscosity. The pronounced difference between the circular polarization of the dimers in fluid media and that of subchloroplast particles and chloroplasts indicates that the former are not suitable models for associated chlorophyll in native structures in electronically excited states. Impairment of the photochemical activity of chloroplasts by heat treatment is accompanied by a reduction of the circular polarization of the fluorescence, which probably reflects a disorganization in structure. The same extent of circular polarization was observed in the fluorescence of chloroplasts regardless whether the reaction centers are open or closed; thus either the same molecules are emitting in the two cases or, if different molecules emit, they are packed in a similar way.

Binding Sites↗