Biomedical subjects

B Grimm

Publications and source records attributed to B Grimm.

At least 19 recordsLinked to original sources

PubMed2005

Periprosthetic fractures around cementless hydroxyapatite-coated femoral stems.

We studied 14 periprosthetic femoral fractures out of a series of 619 hydroxyapatite coated hip implants and compared the outcome to published treatment algorithms using the Vancouver classification. There were five type A fractures, six B1, two B2, and one type B3 fracture. All but one type A fractures were treated conservatively. Compared with the Vancouver classification, we observed a different fracture type in the type B fractures. No fractures at the tip of the stem were seen, as in cemented implants. Three B1 fractures were treated operatively due to fracture displacement, and three were treated conservatively. The B2 and B3 fractures were managed with long, uncemented, revision stems because of a disrupted bone-prosthesis interface. All fractures healed well. This study confirms that the modified algorithm of management of periprosthetic fractures, using the Vancouver classification, is a simple, reproducible, classification system for uncemented prostheses. Conservative treatment is a valid option if the implant is stable whilst surgical intervention is mandatory if the implant is loose.

Aged↗

PubMed2004

Fetal MCG and fetal MEG measurements with a 3-channel SQUID system.

Since the high costs of common large array SQUID system may hinder widespread application of fetal magnetoencephalography (fMEG) and magnetocardiography (fMCG), we intended to investigate a small non-commercial 3-channel SQUID system. The system comprises 3 axial first order gradiometers with 7 cm base length, 2 cm diameter and 2x2 windings of niobium wire, dc-SQUIDs (UJ-111), and current locked mode SQUID electronics that form an equal length triangle (22.5 mm). The system is mounted in a Cryostat BFH-7 model 16 with 5 mm "warm"-"cold" distance. System noise is about 10 fT/Hz1/2. The fMEG and fMCG were recorded between 29 - 40 weeks of gestation after sonographic localization of the fetal head and heart using a 31-channel biomagnetometer (Philips) and the 3-channel-system, both in the same magnetically shielded room. The fMEG was recorded continuously over 500 sec (500 auditory stimuli, 100 dB SPL, 500 Hz, 50 ms, ISI 0.8-1.2/1.6-2.4 sec, trigger channel, maternal ECG lead, sampling rate 1 kHz). The fMCG was recorded over a period of 5 minutes after dewar readjustment. The detection rates of cortical auditory evoked responses (CAER) reached 100 % for both systems. Cross confirmation of the components was difficult and may have uncovered false positive component detection. The fMCG was characterized by a systematic increase in SNR under application of the smaller device. The small size array provides a profitable alternative for the fetal applications.

Cardiotocography↗

PubMed2002

Administration of recombinant rhesus interleukin-12 during acute simian immunodeficiency virus (SIV) infection leads to decreased viral loads associated with prolonged survival in SIVmac251-infected rhesus macaques.

The ability of recombinant rhesus interleukin-12 (rMamu-IL-12) administration during acute simian immunodeficiency virus SIVmac251 infection to influence the quality of the antiviral immune responses was assessed in rhesus macaques. Group I (n = 4) was the virus-only control group. Group II and III received a conditioning regimen of rMamu-IL-12 (10 and 20 microg/kg, respectively, subcutaneously [s.c.]) on days -2 and 0. Thereafter, group II received 2 microg of IL-12 per kg and group III received 10 microg/kg s.c. twice a week for 8 weeks. On day 0 all animals were infected with SIVmac251 intravenously. While all four group I animals and three of four group II animals died by 8 and 10 months post infection (p.i.), all four group III animals remained alive for >20 months p.i. The higher IL-12 dose led to lower plasma viral loads and markedly lower peripheral blood mononuclear cell and lymph node proviral DNA loads. During the acute viremia phase, the high-IL-12-dose monkeys showed an increase in CD3(-) CD8 alpha/alpha(+) and CD3(+) CD8 alpha/alpha(+) cells and, unlike the control and low-IL-12-dose animals, did not demonstrate an increase in CD4(+) CD45RA(+) CD62L(+) naive cells. The high-IL-12-dose animals also demonstrated that both CD8 alpha/alpha(+) and CD8 alpha/beta(+) cells produced antiviral factors early p.i., whereas only CD8 alpha/beta(+) cells retained this function late p.i. Long-term survival correlated with sustained high levels of SIV gag/pol and SIV env cytotoxic T lymphocytes and retention of high memory responses against nominal antigens. This is the first study to demonstrate the capacity of IL-12 to significantly protect macaques from SIV-induced disease, and it provides a useful model to more precisely identify correlates of virus-specific disease-protective responses.

Animals↗

PubMed2002

Subsidence in impaction grafting: the effect of adding a ceramic bone graft extender to bone.

The incidence of revision total hip arthroplasty is increasing dramatically and the associated demand for allograft bone is likely to exceed the available supply. In addition, allograft presents potential problems with regard to infection, antigenicity, availability, reproducibility and cost. It is therefore desirable to develop an alternative to allograft. This study investigated BoneSave, a porous tricalcium phosphate-hydroxyapatite ceramic for use in impaction grafting of the femur at revision total hip arthroplasty. The findings of an in vitro mechanical study comparing the initial stability of pure allograft, a volume mixture of 50 per cent allograft and 50 per cent BoneSave, and a volume mixture of 10 per cent allograft and 90 per cent BoneSave are reported. The BoneSave-allograft mixtures exhibit both much greater mechanical stability and reproducibility than the pure allograft (p < 0.05) at all tested loads (200-800 N). At high peak loads the high volume (90%, v/v) BoneSave mix also provided higher mechanical stability than the medium volume (50 per cent BoneSave-50 per cent allograft) mix (p < 0.05). These results demonstrate thatfrom a mechanical standpoint the tested ceramic provides adequate initial stability to be used as a bone graft extender with allograft in impaction grafting of the femur.

Animals↗

PubMed2001

Tobacco uroporphyrinogen-III decarboxylase: characterization, crystallization and preliminary X-ray analysis.

Uroporphyrinogen-III decarboxylase from Nicotiana tabacum is a plastidial enzyme involved in the biosynthesis of chlorophyll and haem. Sedimentation equilibrium with protein producing diffracting crystals clearly indicates that the enzyme is a homodimer under similar ionic strength conditions to those found in the chloroplast stroma. Additionally, dynamic light scattering reveals an ionic strength dependence for this oligomerization state. Crystals were obtained in the hexagonal space group P622 with one molecule per asymmetric unit and diffracted to 2.3 A resolution using synchrotron radiation.

Crystallization↗

PubMed2001

Crystal structure and substrate binding modeling of the uroporphyrinogen-III decarboxylase from Nicotiana tabacum. Implications for the catalytic mechanism.

The enzymatic catalysis of many biological processes of life is supported by the presence of cofactors and prosthetic groups originating from the common tetrapyrrole precursor uroporphyrinogen-III. Uroporphyrinogen-III decarboxylase catalyzes its conversion into coproporphyrinogen-III, leading in plants to chlorophyll and heme biosynthesis. Here we report the first crystal structure of a plant (Nicotiana tabacum) uroporphyrinogen-III decarboxylase, together with the molecular modeling of substrate binding in tobacco and human enzymes. Its structural comparison with the homologous human protein reveals a similar catalytic cleft with six invariant polar residues, Arg(32), Arg(36), Asp(82), Ser(214) (Thr in Escherichia coli), Tyr(159), and His(329) (tobacco numbering). The functional relationships obtained from the structural and modeling analyses of both enzymes allowed the proposal for a refined catalytic mechanism. Asp(82) and Tyr(159) seem to be the catalytic functional groups, whereas the other residues may serve in substrate recognition and binding, with Arg(32) steering its insertion. The crystallographic dimer appears to represent the protein dimer under physiological conditions. The dimeric arrangement offers a plausible mechanism at least for the first two (out of four) decarboxylation steps.

Amino Acid Sequence↗

PubMed2001

The application of fetal magnetocardiography (FMCG) to investigate fetal arrhythmias and congenital heart defects (CHD).

OBJECTIVES: Fetal magnetocardiography (FMCG), a new non-invasive diagnostic tool in the analysis of the electrophysiological changes of the heart, was selectively applied in cases of fetal arrhythmias and congenital heart defect (CHD) to demonstrate its value for diagnosis and prenatal management. METHODS: The FMCG was analysed and compared to the postnatal ECG in four cases of fetal arrhythmia [supraventricular tachycardia (two cases), complex tachy-/bradycardia (one case), ventricular extrasystoles (one case)] and a case of right heart hypoplasia diagnosed by established methods prior to investigation. RESULTS: A Wolf-Parkinson-White (WPW) syndrome was diagnosed by its characteristic features and the appropriate transplacental therapy chosen. The types of arrhythmia could be characterised in accordance with postnatal ECG findings and irregular conduction was demonstrated in association with a CHD. CONCLUSIONS: The use of the FMCG provides additional information to the common diagnostic tools that influence therapeutic decisions and thus contributes to optimal pre- and postnatal management.

Adult↗

PubMed2001

Loss of alpha-tocopherol in tobacco plants with decreased geranylgeranyl reductase activity does not modify photosynthesis in optimal growth conditions but increases sensitivity to high-light stress.

The enzyme geranylgeranyl reductase (CHL P) catalyses the reduction of geranylgeranyl diphosphate to phytyl diphosphate in higher-plant chloroplasts and provides phytol for both chlorophyll (Chl) and tocopherol synthesis. The reduction in CHL P activity in transgenic tobacco (Nicotiana tabacum L.) plants is accompanied by the reduction in total Chl and tocopherol content and the accumulation of geranylgeranylated Chl (ChlGG). The photosynthetic performance and the susceptibility to photo-oxidative stress have been investigated in these transgenic plants. The reduced total Chl content in Chl P antisense plants resulted in the reduction of electron transport chains per leaf area without a concomitant effect on the stoichiometry, composition and activity of both photosystems. However, Chl P antisense plants were much more sensitive to light stress. Analyses of Chl fluorescence quenching indicated an increased photoinhibitory quenching at the expense of the pH-dependent fluorescence quenching after short illumination (15 min) at moderate light intensities. Prolonged illumination (up to 1 h) at saturating light intensities induced an increased photoinactivation from which the Chl P antisense plants could not recover or could only partially recover during a subsequent low light phase. Our data imply that the presence of ChlGG has no influence on harvesting and transfer of light energy in either photosystem. However, the reduced tocopherol content of the thylakoid membrane is a limiting factor for defensive reactions to photo-oxidative stress.

Chlorophyll↗

PubMed2001

Regulatory network of tetrapyrrole biosynthesis--studies of intracellular signalling involved in metabolic and developmental control of plastids.

Plant tetrapyrrole metabolism is located in two different organelles and distributes end products into the whole cell. A complex regulatory network is involved to prevent metabolic imbalance and inefficient allocation of intermediates as well as to correlate the metabolic activities with organelle development. This review presents new findings about the control of tetrapyrrole biosynthesis and addresses the question of which regulatory principles are involved in controlling the expression of the participating enzymes and the metabolic flow in the entire pathway. It is suggested that functional organelles are required for nuclear gene expression and that metabolic signals participate in a signalling cascade transferring information from plastids to the nucleus. Recent reports about plastid-localised control mechanisms for plant tetrapyrrole metabolism are summarised and compared with results obtained in experiments on nucleus-plastid communication.

5-Aminolevulinate Synthetase↗

PubMed2001

Characterization of Chlamydomonas mutants defective in the H subunit of Mg-chelatase.

Two chlorophyll-deficient mutants of Chlamydomonas reinhardtii, chl1 and brs-1, are light sensitive and, when grown heterotrophically in the dark, accumulate protoporphyrin IX and exhibit yellow/orange pigmentation. The lesions in both mutants were mapped to the gene (CHLH) for the plastid-localized H subunit of the heterotrimeric magnesium chelatase that catalyzes the insertion of magnesium into protoporphyrin IX. The genetic defects in the mutants could be assigned to +1 frameshift mutations in exon 9 (chl1) and exon 10 (brs-1) of the CHLH gene. In both mutants, the H subunit of magnesium chelatase was undetectable, but, as shown for chl1, the steady-state levels of the I and D subunits were unaltered in comparison to wild type. The CHLH gene exhibits marked light inducibility: levels of both the mRNA and the protein product are strongly increased when cultures are shifted from from the dark into the light, suggesting that this protein may play a crucial role in the light regulation of chlorophyll biosynthesis.

Amino Acid Sequence↗

PubMed2001

Chronic immune stimulation accelerates SIV-induced disease progression.

The contribution of chronic immune stimulation on the progression of lentivirus-induced disease was evaluated in the SIVmac251 macaque model of AIDS. Following SIV inoculation, seroconversion and control of the acute viral replication phase, repeated immune stimulations with tetanus toxoid (TT), keyhole limpet hemocyanin (KLH) and allogeneic peripheral blood mononuclear cells (PBMC) were initiated in four monkeys. These animals showed a significant shortening of survival when compared with eight non-immune-stimulated control animals inoculated with the same route, dose and stock of SIVmac251 (median survival 9.5 months versus 17 months, P = 0.010). In addition, when the comparison was extended to another 22 control animals of different origin but inoculated by the same route with similar doses and stocks of SIVmac251, the difference in survival was still significant (9.5 versus 18 months, P = 0.003). This accelerated progression of symptomatic disease was not accompanied with significant increases in plasma viral loads, but suboptimal antibody responses to the immunizing antigens were noted, correlating with the length of survival. These findings may have implications for HIV-infected humans suffering from chronic infectious diseases.

Animals↗

PubMed2001

Extensive developmental and metabolic alterations in cybrids Nicotiana tabacum (+ Hyoscyamus niger) are caused by complex nucleo-cytoplasmic incompatibility.

The genetic basis of multiple phenotypic alterations was studied in cell-engineered cybrids Nicotiana tabacum (+ Hyoscyamus niger) combining the nuclear genome of N. tabacum, plastome of H. niger and recombinant mitochondria. The plants possess a complex, maternally inheritable syndrome of nucleo-cytoplasmic incompatibility, severely affecting growth, metabolism and development. In vivo, the syndrome was manifested as: late germination of seeds; dramatic decrease of chlorophyll and carotenoids in cotyledons and leaves; altered morphology of cotyledons, leaves and flowers; and dwarfism. The leaf phenotype depended on light intensity. In 'green flowers' (an extreme phenotype), homeotic function B was downregulated. In vitro, the incompatibility syndrome was restricted to the pigment deficiency of cotyledons. Electron microscopy revealed perturbations in the differentiation of chloroplasts and palisade parenchyma cells in bleached leaves. The pigment deficiency accompanied by retarded growth is discussed as a result of plastome-genome incompatibility, whereas other features are likely to be due to nucleo-mitochondrial incompatibilities.

Cell Fusion↗

PubMed2001

Impaired expression of the plastidic ferrochelatase by antisense RNA synthesis leads to a necrotic phenotype of transformed tobacco plants.

Protoporphyrin IX is the last common intermediate of tetrapyrrole biosynthesis. The chelation of a Mg2+ ion by magnesium chelatase and of a ferrous ion by ferrochelatase directs protoporphyrin IX towards the formation of chlorophyll and heme, respectively. A full length cDNA clone encoding a ferrochelatase was identified from a Nicotiana tabacum cDNA library. The encoded protein consists of 497 amino acid residues with a molecular weight of 55.4 kDa. In vitro import of the protein into chloroplasts and its location in stroma and thylakoids confirm its close relationship to the previously described Arabidopsis thaliana plastid-located ferrochelatase (FeChII). A 1700-bp tobacco FeCh cDNA sequence was expressed in Nicotiana tabacum cv. Samsun NN under the control of the CaMV 35S promoter in antisense orientation allowing investigation into the consequences of selective reduction of the plastidic ferrochelatase activity for protoporphyrin IX channeling in chloroplasts and for interactions between plastidic and mitochondrial heme synthesis. Leaves of several transformants showed a reduced chlorophyll content and, during development, a light intensity-dependent formation of necrotic leaf lesions. In comparison with wild-type plants the total ferrochelatase activity was decreased in transgenic lines leading to an accumulation of photosensitizing protoporphyrin IX. Ferrochelatase activity was reduced only in plastids but not in mitochondria of transgenic plants. By means of the specifically diminished ferrochelatase activity consequences of the selective inhibition of protoheme formation for the intracellular supply of heme can be investigated in the future.

Cloning, Molecular↗

PubMed2000

Comparative analysis of the low molecular weight and enzymatic antioxidants in response to the phototoxicity of accumulating uroporphyrin and protochlorophyllide in barley leaves treated with cesium chloride.

Cesium chloride treatment of illuminated barley leaves leads to accumulation of uroporphyrinogen which is subsequently either oxidised to uroporphyrin in continuous light or converted to protochlorophyllide in darkness [Shalygo et al. (1998) J Photochem Photobiol 42: 151-158]. We were interested to elucidate the differences in the phototoxicity of uroporphyrin and protochlorophyllide in the CsCI-treated leaves. Photosensitization and the induction of oxidative stress responses in the barley leaves occurred much faster upon protochlorophyllide than upon uroporphyrin accumulation. We compared the time resolved changes in the pool sizes of low molecular weight antioxidants, such as ascorbate, glutathione and tocopherol, as well as of the enzymatic activities of catalase, ascorbate peroxidase, glutathione reductase and superoxide dismutase in illuminated barley leaves which accumulated uroporphyrin or protochlorophyllide. A rapid loss of the antioxidant levels correlated with the accumulation of reactive oxygen species. The contents of low molecular weight antioxidants and the activities of most of the antioxidative enzymes declined more rapidly in the presence of protochlorophyllide than of uroporphyrin. Due to its high lipophilicity, free protochlorophyllide is associated with biomembranes. Therefore, it is assumed that it exerts its phototoxic effects to membranes more rapidly than uroporphyrin.

Journal Article↗

PubMed2000

Decreased and increased expression of the subunit CHL I diminishes Mg chelatase activity and reduces chlorophyll synthesis in transgenic tobacco plants.

The chelation of Fe2+ and Mg2+ ions forms protoheme IX and Mg-protoporphyrin IX, respectively, and the latter is an intermediate in chlorophyll synthesis. Active magnesium protoporphyrin IX chelatase (Mg-chelatase) is an enzyme complex consisting of three different subunits. To investigate the function of the CHL I subunit of Mg-chelatase and the effects of modified Mg-chelatase activity on the tetrapyrrole biosynthetic pathway, we characterized N. tabacum transformants carrying gene constructs with the Chl I cDNA sequence in antisense and sense orientation under the control of the CaMV 35S promoter. Both elevated and diminished levels of Chl I mRNA and Chl I protein led to reduced Mg-chelatase activities, reflecting a perturbation of the assembly of the enzyme complex. The transformed plants did not accumulate the substrate of Mg-chelatase, protoporphyrin IX, but the leaves contained less chlorophyll and possessed increased chlorophyll a/b ratios, as well as a deficiency of light-harvesting chlorophyll binding proteins of photosystems I and II. The expression and activity of several tetrapyrrolic enzymes were reduced in parallel to lower the Mg-chelatase activity. Consistent with the lower chlorophyll contents, the rate-limiting synthesis of 5-aminolevulinate was also decreased in the transgenic lines analyzed. The consequence of reduced Mg-chelatase on early and late steps of chlorophyll synthesis, and on the organization of light harvesting complexes is discussed.

Aminolevulinic Acid↗

PubMed2000

Overexpression of plastidic protoporphyrinogen IX oxidase leads to resistance to the diphenyl-ether herbicide acifluorfen.

The use of herbicides to control undesirable vegetation has become a universal practice. For the broad application of herbicides the risk of damage to crop plants has to be limited. We introduced a gene into the genome of tobacco (Nicotiana tabacum) plants encoding the plastid-located protoporphyrinogen oxidase of Arabidopsis, the last enzyme of the common tetrapyrrole biosynthetic pathway, under the control of the cauliflower mosaic virus 35S promoter. The transformants were screened for low protoporphyrin IX accumulation upon treatment with the diphenyl ether-type herbicide acifluorfen. Leaf disc incubation and foliar spraying with acifluorfen indicated the lower susceptibility of the transformants against the herbicide. The resistance to acifluorfen is conferred by overexpression of the plastidic isoform of protoporphyrinogen oxidase. The in vitro activity of this enzyme extracted from plastids of selected transgenic lines was at least five times higher than the control activity. Herbicide treatment that is normally inhibitory to protoporphyrinogen IX oxidase did not significantly impair the catalytic reaction in transgenic plants and, therefore, did not cause photodynamic damage in leaves. Therefore, overproduction of protoporphyrinogen oxidase neutralizes the herbicidal action, prevents the accumulation of the substrate protoporphyrinogen IX, and consequently abolishes the light-dependent phytotoxicity of acifluorfen.

Blotting, Northern↗

PubMed2000

Role of magnesium chelatase activity in the early steps of the tetrapyrrole biosynthetic pathway.

Magnesium-protoporphyrin IX chelatase (Mg-chelatase) is located at the branchpoint of tetrapyrrole biosynthesis, at which point protoporphyrin IX is distributed for the synthesis of chlorophyll and heme. We investigated the regulatory contribution of Mg-chelatase to the flow of metabolites. In plants, the enzyme complex consists of three subunits, designated CHL D, CHL I, and CHL H. Transgenic tobacco (Nicotiana tabacum) plants expressing antisense RNA for the Mg-chelatase subunit CHL H were analyzed to elucidate further the role of Mg-chelatase in the distribution of protoporphyrin IX into the branched tetrapyrrolic pathway. The transgenic plants displayed a reduced growth rate and chlorophyll deficiency. Both phenotypical properties were correlated with lower Mg-chelatase activity. Unexpectedly, less protoporphyrin IX and heme accumulated, and a decrease in 5-aminolevulinate (ALA)-synthesizing capacity and ALA dehydratase activity paralleled the progressive reduction in Mg-chelatase activity in the transformants compared with control plants. The reduced activities of the early enzymatic steps corresponded with lower levels of transcripts encoding glutamyl-tRNA reductase and ALA-dehydratase. The decreased expression and activities of early enzymes in the pathway could be explained by a feedback-controlled mechanism in response to lower Mg-chelatase activity. We discuss intercompartmental signaling that synchronizes the activities of the first steps in tetrapyrrolic metabolism with the late steps for the synthesis of end products.

Lyases↗

PubMed1999

Mg-chelatase of tobacco: the role of the subunit CHL D in the chelation step of protoporphyrin IX.

The Mg-chelation is found to be a prerequisite to direct protoporphyrin IX into the chlorophyll (Chl)-synthesizing branch of the tetrapyrrol pathway. The ATP-dependent insertion of magnesium into protoporphyrin IX is catalyzed by the enzyme Mg-chelatase, which consists of three protein subunits (CHL D, CHL I, and CHL H). We have chosen the Mg-chelatase from tobacco to obtain more information about the mode of molecular action of this complex enzyme by elucidating the interactions in vitro and in vivo between the central subunit CHL D and subunits CHL I and CHL H. We dissected CHL D in defined peptide fragments and assayed for the essential part of CHL D for protein-protein interaction and enzyme activity. Surprisingly, only a small part of CHL D, i.e., 110 aa, was required for interaction with the partner subunits and maintenance of the enzyme activity. In addition, it could be demonstrated that CHL D is capable of forming homodimers. Moreover, it interacted with both CHL I and CHL H. Our data led to the outline of a two-step model based on the cooperation of the subunits for the chelation process.

Chlorophyll↗