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Biomedical subjects

I Schmidt

Publications and source records attributed to I Schmidt.

At least 55 records · Page 3Linked to original sources
PubMed1997

The leptin receptor mediates apparent autocrine regulation of leptin gene expression.

The possibility that the leptin receptor (LEPR) mediates autocrine regulation of leptin expression in adipose tissue was examined in 10-day-old Zucker rat pups with different copy numbers of the leptin receptor mutation (Lepr(fa)). Plasma leptin concentrations and adipose tissue mRNA levels for leptin were related to copy number of the mutation (fa/fa > fa/+ > +/+). These relationships were independent of plasma insulin concentration. Reduced copy number for the functional leptin receptor apparently results in a diminished negative feedback signal to the leptin gene in adipose tissue. Thus, leptin appears to close a short regulatory loop controlling its own synthesis in adipose tissue.

Adipose Tissue↗
PubMed1997

Studies on spontaneous promoter-up mutations in the transcriptional activator-encoding gene phIR and their effects on the degradation of phenol in Escherichia coli and Pseudomonas putida.

The activator-encoding gene phlR was identified upstream of the plasmid-encoded operon for phenol degradation in Pseudomonas putida strain H by cassette mutagenesis and DNA sequence analysis. The deduced amino acid sequence of PHLR shows high homology to DmpR of P. putida sp. CF600 and to the chromosomally encoded PhhR of P. putida P35X reported previously. Trans-activation of phenol degradation was observed when phlR was overexpressed in a phlR insertion mutant. Transconjugants of Escherichia coli carrying pPGH11, which contains the complete set of phl genes, are unable to grow on phenol as carbon source. However, two types of mutants were selected for further characterization that were able to metabolize phenol as sole source of carbon and energy. In both types of mutants enhanced expression of phlR is responsible for the Phl+ phenotype. In type I (pPGH13) a deletion of 1 bp made the -35 region and the spacing between the -35 and -10 regions of the phlR promoter more similar to the consensus structure. In type II (pPGH14) a duplication of the phlR 5' region was identified that includes part of the -35 motif and reduces the spacing between the -35 and -10 regions. In addition, due to the duplication of part of phlR, the distance from the phlR promoter to the catabolic phl operon is increased. Different transcriptional start sites have been identified by primer extension analysis in clones harboring pPGH14 or the wild type phlR. Quantitative primer extension analysis revealed that the greatest amount of phlR transcript is expressed from the partial, phlR duplication. Growth on phenol and phenol hydroxylase activity reflect the high level of phlR transcript in E. coli transconjugants. Overexpression of PhlR was also observed when pPGH14 was transferred into P. putida, and results in earlier induction of the phenol degradation operon relative to the wild-type strain.

Bacterial Proteins↗
PubMed1997

Leptin does not reduce body fat content but augments cold defense abilities in thermoneutrally reared rat pups.

A previous study showed that recombinant leptin markedly affects the body fat content and thermoregulatory energy expenditure of lean (+/+ and +/fa) suckling-age rats, and we wanted to find out whether leptin in doses that halved body fat of cold-reared lean pups had any effect in thermoneutrally reared lean pups. When +/+ pups were artificially reared from 4 to 16 days of age at thermoneutrality and treated as before with leptin from day 7, their total metabolic rate throughout the treatment period was only 4% higher than that of the control littermates and their final body fat content only 4% lower (both P>0.05). We conclude from comparisons of the results in +/+ pups at cold and thermoneutral conditions that leptin does not stimulate, but rather disinhibits, sympathetically mediated thermoregulatory thermogenesis.

Adipose Tissue↗
PubMed1997

Postnatal changes in mitochondrial protein mass and respiration in skeletal muscle from the newborn pig.

Quantitative and functional changes occurring in mitochondria were studied in pig skeletal muscle between birth and 5 days of life. Postnatal changes were followed separately on intermyofibrillar and subsarcolemmal mitochondria isolated from rhomboïdeus (RH) and longissimus dorsi (LD) muscles. The integrity and purity of the isolated mitochondria was checked by electron microscopic observations. The mass of mitochondrial protein was not different between muscles at birth. It increased tremendously during the first 5 days of life, by 49% in LD (P < 0.001) and 93% in RH (P < 0.001) muscle and was 30% higher in RH than in LD muscle at 5 days of life (P < 0.05). Mitochondria isolated from RH muscle exhibited 30% higher oxidative and phosphorylative capacities than those from LD muscle at 5 days of life (P < 0.05). Intermyofibrillar (IM) mitochondria had high respiration rate, enzyme activities and coupling parameters (respiratory control ratio, phosphorus-oxygen ratio) from birth. Subsarcolemmal (SS) mitochondria were less active than IM mitochondria; their respiration rate and enzyme activities were 60% lower (P < 0.01) and increased with age, particularly in LD muscle (P < 0.05). Short-term cold exposure had no effect on mitochondrial mass and activity. These results suggest that muscle mitochondria are functional from birth and are changing primarily quantitatively. SS and IM mitochondria exhibit specific changes that are probably involved in the postnatal acquisition of skeletal muscle oxidative metabolism.

Animals↗
PubMed1997

Positive selection and transplantation of peripheral CD34+ progenitor cells: feasibility and purging efficacy in pediatric patients with neuroblastoma.

Peripheral stem cells were mobilized and collected in 20 children with stage 4 neuroblastoma. A total of 37 leukaphereses were performed in the 20 patients. The mean number of collected cells was 5.6 +/- 2.4 x 10(8)/kg (range 1.9-10.5), and the number of collected CD34+ progenitors was 6.1 +/- 6.3 x 10(6)/kg (range 0.75-21.7). CD34-positive selection was performed using the CellPro method. Of the adsorbed cells, 42 +/- 20% (range 4.3-76.6) stained positively for CD34, and the number of positively selected CD34+ cells was 2.0 +/- 1.9 x 10(6)/kg (range 0.09-7.1). The mean recovery of CD34+ cells was 36 +/- 20% (range 6-67). For detection of contaminating neuroblastoma cells before and after CD34-positive selection, a murine antidisialoganglioside GD2 antibody (14.G2a) was used, followed by the alkaline phosphatase antialkaline phosphatase (APAAP) method. Before the positive selection, various numbers of contaminating neuroblastoma cells were found in the leukaphereses of 7 patients. After positive selection, neuroblastoma cells were still detectable in all 7 patients, with a mean log depletion of tumor cells of 1.41 +/- 0.45 (range 0.69-2.13). In 1 patient, contaminating neuroblastoma cells were found only after CD34-positive selection. In 15 of the 20 patients, high-dose chemotherapy was performed, and positively selected CD34+ cells were reinfused in 12 patients. In 10 of these, the mean time to reach > 0.5 x 10(9)/L granulocytes was 12.3 +/- 1.7 days (range 10-16). One patient died at day 7 due to sepsis, and in 1 patient the backup was given at day 15. Because of the low number of collected CD34+ cells, 3 patients were grafted with a combination of unmanipulated PBSC and CD34+ progenitors. In summary, we have shown that positive selection of peripheral CD34+ progenitors is feasible in pediatric patients. However, strategies to improve the recovery of the CD34+ cells and the purging efficacy of this method (i.e., higher enrichment of CD34+ cells, combination of positive and negative selection methods) should be evaluated further.

Animals↗
PubMed1997

Different physiological traits underlying increased body fat of fatty (fa/fa) and heterozygous (+/fa) rats.

To find out whether the most characteristic physiological traits distinguishing suckling-age fa/fa pups from lean littermates also differ between +/+ and +/fa littermates, we analyzed the body composition and cold defense of 7- and 16-day-old pups and the plasma concentrations of insulin, glucose, triglycerides, and free fatty acids in 16-day-old pups. Zucker rat x Brown Norway hybrid pups were genotyped by using a molecular marker within 0.5 cM of the fa gene. At both ages the +/fa pups had significantly more body fat than their +/+ littermates. At 7 days this difference was as large as that between +/fa and fa/fa pups, but at 16 days it was only one-seventh of the fa/fa vs. +/fa difference. In contrast, there were no heterozygote differences for three parameters that show crucial abnormalities in the fa/fa pups: thermoregulatory thermogenesis and plasma concentrations of insulin and triglycerides. The physiological mechanisms underlying the increased fat content of +/fa pups thus differ from those known to fuel most of the excessive fat deposition of their fa/fa littermates.

Adipose Tissue↗
PubMed1997

[The treatment concept in severe hand injuries].

Treatment of complex injuries of the hand aims at best possible reconstruction of function and quick social and professional rehabilitation. This objective is hardly to achieve in most cases due to the severity of injury. Outcome depends on patients' age and injury pattern and essentially on the strategy of surgical treatment. Errors in prehospital emergency treatment can hardly be corrected, no matter how excellent the surgical technique may be. Traumatic multiple finger or thumb amputations are clear indications for replantation. Beginning with osteosynthesis, suture of tendons and ligaments the restoration of blood supply is the most important step. Nerve reconstruction should be performed immediately, but if tension free repair is impossible, secondary nerve grafting is preferred. Primary skin closure should be carried out if possible, in case of defects skin grafts, local or free flaps have to be used.

Amputation, Traumatic↗
PubMed1996

The aphid transmission factor of cauliflower mosaic virus forms a stable complex with microtubules in both insect and plant cells.

We analyzed the distribution of the cauliflower mosaic virus (CaMV) aphid transmission factor (ATF), produced via a baculovirus recombinant, within Sf9 insect cells. Immunogold labeling revealed that the ATF colocalizes with an atypical cytoskeletal network. Detailed observation by electron microscopy demonstrated that this network was composed of microtubules decorated with paracrystalline formations, characteristic of the CaMV ATF. A derivative mutant of the ATF, unable to self-assemble into paracrystals, was also analyzed. This mutant formed a net-like structure, with a mesh of four nanometers, tightly sheathing microtubules. Both the ATF- and the derivative mutant-microtubule complexes were highly stable. They resisted dilution-, cold-, and calcium-induced microtubule disassembly as well as a combination of all three for over 6 hr. CaMV ATF cosedimented with microtubules and, surprisingly, it bound to Taxol-stabilized microtubules at high ionic strength, thus suggesting an atypical interaction when compared with that usually described for microtubule-binding proteins. Using immunofluorescence double labeling we also demonstrated that the CaMV ATF colocalizes with the microtubule network when expressed in plant cells.

Animals↗
PubMed1996

Apoptotic cell death in activated monocytes following incorporation of clodronate-liposomes.

The present study was performed to elucidate whether sterically stabilized liposomes laden with clodronate, which lead to depletion of macrophages (Mphis) and amelioration of experimental autoimmune arthritis in vivo, selectively affect cells of the mphi lineage in vitro. The rates of incorporation of drug-free, fluorescent liposomes and the rates of cell death following exposure to clodronate-liposomes were assessed in human peripheral blood monocytes, as well as in polymorphonuclear leukocytes (PMNs), T cells, endothelial cells, and fibroblasts, both at rest and following activation. Gel electrophoresis of nuclear extracts and ultrastructural analyses were performed to identify the modality of cell death. Monocytes, particularly upon activation, were more efficient in incorporating sterically stabilized liposomes than all other cells except PMNs. Twenty percent of resting monocytes and up to 65% of activated monocytes died within 24 h of exposure to clodronate-liposomes, whereas the other cell types, including PMNs, remained unaffected. Activated monocytes exposed to clodronate-liposomes, but not resting or activated monocytes exposed to drug-free liposomes, showed clear signs of apoptotic cell death. In most of the assays, sterically stabilized liposomes were more efficient than conventional phosphatidylcholine-liposomes. Sterically stabilized clodronate-liposomes preferentially affect cells of the mphi lineage, particularly if activated. Selective elimination of activated Mphis by apoptosis may explain both therapeutic efficacy and safety of clodronate-liposomes in experimental models of autoimmunity.

Analgesics, Non-Narcotic↗
PubMed1996

Improved traceability of pH measurements.

Traceability is a prerequisite for the comparability and uniformity of measurements. Although pH-measurements are carried out on a large scale in laboratory and industry, the problems involved in the traceability of pH values have not adequately been solved in the past. The comparability of pH measurements is limited, among other parameters, by the accuracy of the pH values of the standard buffer solutions used to calibrate the pH meter-electrode assemblies. The measured pH(X) value must be traceable to primary standard pH(PS) values through an unbroken chain of comparisons, all values having stated uncertainties. A new primary standard measurement device for pH is used to certify primary pH reference materials from which these secondary reference materials can be derived.

Journal Article↗
PubMed1996

Recovery of speed of information processing in closed-head-injury patients.

After severe traumatic brain injury, patients almost invariably demonstrate a slowing of reaction time, reflecting a slowing of central information processing. Methodological problems associated with the traditional method for the analysis of longitudinal data (MANOVA) severely complicate studies on cognitive recovery. It is argued that multilevel models are often better suited for the analysis of improvement over time in clinical settings. Multilevel models take into account individual differences in both overall performance level and recovery. These models enable individual predictions for the recovery of speed of information processing. Recovery is modelled in a group of closed-head-injury patients (N = 24). Recovery was predicted by age and severity of injury, as indicated by coma duration. Over a period up to 44 months post trauma, reaction times were found to decrease faster for patients with longer coma duration.

Adolescent↗
PubMed1996

Perinatal energy stores and excessive fat deposition in genetically obese (fa/fa) rats.

To find out whether perinatal differences in energy stores are essential for the phenotypic manifestation of the fa/fa genotype in 16-day-old rats, we investigated Zucker rats (13M) x Brown Norway (BN) hybrids. +/fa and fa/fa hybrids were identified by molecular polymorphisms approximately 1 cM from the fa locus. The fat content of all pups increased rapidly after birth, and differences between +/fa and fa/fa hybrid pups were significant in 7- and 16-day-old rats but not at 0, 1, and 4 days of age. For 16-day-old 13MBN hybrids as well as 13M Zucker rats, plots of carcass fat vs. body mass were closely fitted by parallel regression lines, with carcass fat similarily higher in fa/fa than +/fa pups of both strains. Fetal and neonatal plasma insulin concentration and hepatic glycogen in the 13MBN hybrid pups did not differ between +/fa and fa/fa litter-mates, nor did the hepatic and adipose tissue triglycerides in 24-h-old 13MBN hybrid pups. The manifest differences between fatty and lean 16-day-old pups thus do not depend on differences in perinatal energy stores.

Adipose Tissue↗
PubMed1996

Is the onset of obesity in suckling fa/fa rats linked to a potentially larger milk intake?

We wanted to find out whether fatty (fa/fa) sucklings show abnormal intake when given access to an abundant milk reservoir. To do this, we gravimetrically determined the milk ingested by small groups (4-5 pups) of 5- to 15-day-old lean (+/fa) and fatty littermates allowed to suckle for 30 min after their mother had not been nursing for periods of between 1 and 7 h. The pups were grouped randomly and their phenotypes retrospectively identified. Within both genotypes, the intakes of simultaneously tested pups were significantly higher in pups deprived for longer periods. Deprived and undeprived fa/fa pups ingested, however, slightly but significantly less milk than +/fa littermates did in the same nursing bout. In the first 2 wk of life, when fa/fa pups deposit nearly twice as much body fat as their +/fa littermates do, fa/fa pups will thus suckle less rather than more milk. This extends previous findings showing that the onset of fa/fa obesity is independent of larger intakes and thus questions that fa impairs a receptor primarily controlling food intake.

Animals↗
PubMed1996

Manipulation of potential perinatal zeitgebers for the juvenile circadian temperature rhythm in rats.

We investigated the importance of pre- and postnatal maternal rhythmicity for the development and synchronization of the juvenile circadian core temperature (Tc) rhythm in rats by evaluating the Tc of artificially reared pups in six litters derived from mothers maintained in continuous bright light (LL) and impregnated after drinking behavior stopped showing circadian periodicity. Pups removed from their aperiodic mothers on postnatal day 9 and artificially reared for 3 days showed a free-running Tc rhythm whose amplitude (3.1 +/- 0.1 degrees C, n = 47) was only slightly smaller than that of control pups born to mothers maintained in a 12:12-h light-dark cycle. In four litters the acrophases were not much less synchronized than in the four control litters (mean vector lengths 0.79 +/- 0.05 vs. 0.94 +/- 0.04). In two litters synchronization among littermates was not significant. Additional experiments with cross-fostered pups showed that synchronization is not caused by the time of birth. We conclude that synchronization among littermates can develop even when maternal rhythmicity has been suppressed even before conception, possibly because the Tc decrease of several littermates happening to have similar phases at birth acts as zeitgeber.

Animals↗
PubMed1996

Leptin reduces juvenile fat stores by altering the circadian cycle of energy expenditure.

The recently identified hormone leptin (ob protein) secreted by white adipose tissue is widely thought to provide a feedback signal limiting fat storage by decreasing food intake. By artificially rearing leptin-treated and control littermates fed identical amounts of milk, however, we show here that lean suckling-age rats treated with recombinant murine leptin can reduce fat storage solely by increasing energy expenditure. Continuous measurements of core temperature and metabolic rate show that this increase is not uniform throughout the day but is especially prominent in the morning when rat pups usually conserve energy by entering a torpor-like state. Leptin's alleviation of hypometabolic, torpor-like states is thus not restricted to cases of impaired hormone production but seems instead to be a normal biological function independent of its effects on food intake.

Adipose Tissue↗
PubMed1996

Dimeric lipoprotein lipase is bound to triglyceride-rich plasma lipoproteins.

Lipoprotein lipase hydrolyzes the triglyceride-rich core of chylomicrons and very low density lipoproteins. It is also a ligand, in vitro, for binding of lipoproteins to the low density lipoprotein receptor-related protein and may play a central role in the receptor-mediated removal of triglyceride-rich lipoproteins. The aim of the present study was to determine to which lipoprotein subclass the enzyme is bound in preheparin plasma and when released into plasma by heparin injection. Tetrahydrolipstatin, a potent inhibitor of serine lipases, was used to block lipolytic activity, thereby preventing changes in plasma lipoproteins due to ex vivo lipolysis. To analyze the distribution pattern of lipoprotein lipase dimers among lipoprotein classes, a specific ELISA was used and gel filtration was performed in pre- and postheparin plasma from five subjects with triglyceride ranging from 69 to 522 mg/dl. When lipolytic activity was not inhibited, lipoprotein lipase dimers eluted in association with low and high density lipoproteins, reproducing results previously obtained by several groups of investigators. However, in pre- and postheparin samples treated with tetrahydrolipstatin, most of the dimeric enzyme was found associated with very low density lipoprotein particles. In conclusion in pre- and postheparin samples most of the lipoprotein lipase dimers are associated with very low density lipoproteins when ex vivo lipolytic activity is inhibited, which supports the hypothesis that, in vivo, lipoprotein lipase may affect the receptor-mediated removal of these particles. Moreover, it suggests that the association between lipoprotein lipase and cholesterol-rich lipoproteins might be an ex vivo phenomenon due to lack of inhibition of lipolytic activity.

Adult↗
PubMed1995

Sec-dependent thylakoid protein translocation. Delta pH requirement is dictated by passenger protein and ATP concentration.

A Sec-type system is responsible for the translocation of a subset of proteins across the thylakoid membrane in higher plant chloroplasts. Previous studies have suggested that the thylakoidal delta pH plays a minor role in this translocation mechanism, but we show here that it can be essential for the translocation process, depending on the identity of the passenger protein and the concentration of ATP. Studies using chimeric proteins show that, whereas the presequence dictates the translocation pathway, the delta pH requirement is dictated exclusively by the passenger protein; some passenger proteins are virtually delta pH-independent whereas others are absolutely dependent. delta pH requirement is not related to charge characteristics of the passenger proteins, ruling out an electrophoretic effect. Analysis of the 33-kDa photosystem II protein reveals an inverse relationship between delta pH requirement and ATP concentration; import into isolated thylakoids is inhibited 14-fold by nigericin at moderate ATP concentrations, and totally inhibited when the ATP concentration is reduced to 2 microM. The results indicate that the roles of the delta pH and ATP overlap and suggest that the delta pH may be obligatory when the passenger protein is abnormally difficult to translocate, possibly due to the folding of the polypeptide chain. We compare the energetics of this system with those of prokaryotic systems from which the chloroplast system is believed to have evolved.

Adenosine Triphosphate↗