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S David

Publications and source records attributed to S David.

At least 127 records · Page 7Linked to original sources

The rôle of adsorption in beta 2-microglobulin removal.

BK-PMMA 2.1 m2 surface dialysers were used in vivo and in vitro for evaluation of their capacity for beta 2-M removal by both adsorption and elimination through the membrane. As expected, in vivo, beta 2-M plasma concentrations were more effectively reduced when the dialyser was used in haemofiltration, rather than in haemodialysis. However, the mass recovered in the ultrafiltrate could not account for the observed difference in beta 2-M removal. In vitro tests demonstrated that membrane adsorption of beta 2-M is greater when the dialyser is used in convection than in diffusion. Therefore, the greater beta 2-M removal during haemofiltration depends on both increased adsorption and membrane elimination by convection. Thanks to adsorption, beta 2-M removal by haemodialysis was greater than for other synthetic membranes, but was insufficient to obtain beta 2-M removal corresponding to the estimated generation.

Adsorption↗

Conversion of 5-aminoimidazole ribotide to the pyrimidine of thiamin in enterobacteria: study of the pathway with specifically labeled samples of riboside.

Samples of 5-amino-1-(beta-D-ribofuranosyl)imidazole labeled with 13C at position C-1 or C-2 of the ribose part or with 15N at position N-3 or amino of the imidazole part were prepared by chemical synthesis. The incorporation of label from these samples into the pyrimidine of thiamin biosynthesized by a mutant strain of Salmonella typhimurium was studied by GC-MS. The results show clearly that in enterobacteria the methyl carbon atom and the N-1 nitrogen atom of one molecule of thiamin pyrimidine derive from the same molecule of 5-aminoimidazole ribotide. More specifically, the methyl carbon atom comes from the carbon C-2' of the ribose part and the nitrogen N-1 from nitrogen N-3 of the imidazole; furthermore, the amino nitrogen of the aminoimidazole becomes the amino nitrogen of the pyrimidine.

Carbon Isotopes↗

Preparation of disaccharides having a beta-D-mannopyranosyl group from N-phthaloyllactosamine derivatives by double or triple SN2 substitution.

Condensation of 1,2,3,4,6-penta-O-acetyl-beta-D-galactopyranose with methyl 3,6-di-O-benzyl-2-deoxy-2-phthalimido-beta-D-glucopyranoside, followed by alkaline methanolysis, gave a derivative of lactosamine that has an unsubstituted beta-D-galactopyranosyl group. Tributyltin oxide-mediated allylation gave a good yield of the 3'-O-allyl (9) and a poor yield of the 3',6'-di-O-allyl ether (8). Protection of 9 at O-6' was achieved by reductive opening of the 4',6'-O-anisylidene derivative, to give the 6'-O-(4-methoxybenzyl) ether 15. Conversion of 8 and 15 to their 2',4'-bis(trifluoromethanesulfonates), followed by SN2 reaction with benzoate, gave the corresponding beta-D-mannopyranosyl disaccharides. However, the model methyl 3-O-allyl-beta-D-galactopyranoside and 9 were converted into beta-D-mannopyranosyl derivatives in better yield (52-55%) by a one-pot, triple SN2 substitution of the tris(trifluoromethanesulfonates).

Amino Sugars↗

Coronary artery spasm induced by anesthesia: a case report and review of the literature.

Anesthesia-induced coronary vasospasm has been reported only rarely. We report a case, without previous cardiac history, in which immediately after anesthesia induction a marked ST elevation was noted on the EKG monitor. Premature ventricular contractions as well as non-sustained ventricular tachycardia were noted. These changes resolved immediately after nitroglycerin infusion and 75 mg of lidocaine were given. A coronary angiogram revealed normal coronary arteries and left ventriculogram. Ergonovine stimulation was not performed. The patient was discharged home on calcium entry blockers and nitrates. Exercise stress test two weeks after discharge was negative for ischemia. Induction of anesthesia triggering coronary spasm has been reported rarely, and to our knowledge never in the presence of angiographically normal coronary anatomy. Coronary vasospasm with typical EKG changes--namely, ST elevation and ventricular arrhythmias--has to be included as a possible complication of general anesthesia. Recognition of this syndrome allows prompt treatment and prevention of future episodes.

Anesthesia, General↗

Identification of a cadherin cell adhesion recognition sequence.

The molecular mechanisms by which the cadherins interact with one another to promote cell adhesion have not been elucidated. In particular, the amino acid sequences of the cadherin cell adhesion recognition sites have not been determined. Here we demonstrate that synthetic peptides containing the sequence HAV, which is common to all of the cadherins, inhibit two processes (compaction of eight-cell-stage mouse embryos and rat neurite outgrowth on astrocytes) that are known to be mediated by cadherins. The data suggest that the tripeptide HAV is a component of a cadherin cell adhesion recognition sequence.

Amino Acid Sequence↗

Macrophages can modify the nonpermissive nature of the adult mammalian central nervous system.

Although astrocytic gliosis has been linked to failure of axonal regeneration in the adult mammalian CNS, its role is not fully established. We used an in vitro assay to investigate the role of reactive astrocytes and macrophages in influencing axonal growth in the lesioned adult rat optic nerve. Soon after optic nerve transection, the nonpermissive nature of the optic nerve is altered to a permissive state near the lesion. This may account for injury-induced axonal sprouting and may contribute to the failure of these sprouts to elongate beyond the site of the lesion in vivo. We provide evidence that this lesion-induced change in the axonal growth-promoting properties of the CNS near the lesion may be produced by mononuclear phagocytes. In addition, several months after optic nerve transection, the degenerated nerves, which consist mainly of astrocytes and lack myelin, i.e., astrocytic "scar" tissue, are a good substrate for neurite growth. Taken together, these results suggest that in this in vitro system, substantial inhibitory effects are not associated with regions of astrocytic gliosis and that the nonpermissive nature of the CNS white matter can be modified by macrophages.

Animals↗

A dual laminin/collagen receptor acts in peripheral nerve regeneration.

A regeneration chamber was created in vivo by suturing a synthetic tube sealed at its distal end onto the proximal stump of a severed rat sciatic nerve. Nerves regenerated into tubes coated with laminin at a rate of 0.33 mm/day after a lag of about 2 days. At 25 days, regenerating nerves had extended 23% farther into laminin-coated tubes as compared with uncoated ones. Monoclonal antibody 3A3, which functionally interferes with a dual laminin/collagen receptor, inhibited nerve regeneration into laminin-coated tubes by 32%. In contrast, monoclonal antibody JG22, which inhibits chicken matrix receptors, had no significant effect on regeneration. Immunohistochemical studies of teased adult rat sciatic nerves indicate that 3A3 bound to Schwann cells and possibly to axons. In other studies, the heterodimeric, laminin/collagen receptor recognized by 3A3 has been shown to be a member of the integrin superfamily of adhesive receptors. These data provide evidence that an integrin receptor functions in nerve regeneration in vivo.

Animals↗

Nucleotide sequence and expression in Escherichia coli of the Lactococcus lactis citrate permease gene.

The plasmid-encoded citrate determinant of the Lactococcus lactis subsp. lactis var. diacetylactis NCDO176 was cloned and functionally expressed in a Cit- Escherichia coli K-12 strain. From deletion derivative analysis, a 3.4-kilobase region was identified which encodes the ability to transport citrate. Analysis of proteins encoded by the cloned fragment in a T7 expression system revealed a 32,000-dalton protein band, which correlated with the ability of cells to transport citrate. Energy-dependent [1,5-14C]citrate transport was found with membrane vesicles prepared from E. coli cells harboring the citrate permease-expressing plasmid. The gene encoding citrate transport activity, citP, was located on the cloned fragment by introducing a site-specific mutation that abolished citrate transport and resulted in a truncated form of the 32,000-dalton expression product. The nucleotide sequence for a 2.2-kilobase fragment that includes the citP gene contained an open reading frame of 1,325 base pairs coding for a very hydrophobic protein of 442 amino acids, which shows no sequence homology with known citrate carriers.

Amino Acid Sequence↗

Involvement of interferon-system in the regulation of cell growth and differentiation.

In this report we review the current knowledge on the involvement of the interferon (IFN) system in the regulation of cell growth and differentiation. We also summarize our own data which provide evidence for the strong correlation between IFN-mediated growth-arrest of transformed cells and the elevated enzymatic activity of an IFN-induced protein. Similarly, it is demonstrated that elevated levels of IFN-induced proteins accompany the early phases of in-vitro cell differentiation. IFN-treatment of NIH/3T3 mouse fibroblasts transformed by Moloney-murine sarcoma virus (MSV) resulted in a significant reduction in the rates of cell growth, protein synthesis and cloning efficiency. In parallel, 2-5A-synthetase activity was induced ten-fold above the background level. Treatment of these cells for 3 days with 450 international units (IU)/ml of IFN followed by its removal, resulted in a gradual increase in all parameters associated with cell growth while the 2-5A-synthetase activity was reduced to its normal level. However, almost no recovery occurred when cells were treated with 1,800 IU/ml. In parallel, 2-5A-synthetase activity remained highly elevated even at 3 days after the removal of IFN. In these cells, the expression of both c-myc and v-mos was reduced rapidly following IFN treatment. Upon removal of IFN after 24 h of treatment, the expression of both genes was resumed but with a different kinetics, suggesting that different mechanisms are responsible for the reduction in gene expression. In rat skeletal muscle cultures which differentiate to form myotubes, the level of both 2-5A-synthetase and protein kinase activities was transiently elevated, reaching a peak at 3 days followed by a decrease to background levels. This peak activity precedes the appearance of the major muscle differentiating proteins.

2',5'-Oligoadenylate Synthetase↗

Plasmid transformation by electroporation of Leuconostoc paramesenteroides and its use in molecular cloning.

In this report, we demonstrate the utility of electroporation as an efficient method for genetic transformation of Leuconostoc paramesenteroides. We optimized several factors which determine the transformation frequency, resulting in transformation efficiencies of up to 4 x 10(3) transformants per micrograms of pNZ12 DNA, which contains the promiscuous Lactococcus lactis pSH71 replicon. Slightly lower efficiencies were obtained with a deletion derivative of the broad-host-range plasmid pAM beta 1. These plasmids could be stably maintained in L. paramesenteroides NZ6009 for more than 100 generations, even in the absence of selective pressure. In order to show the use of the developed host-vector system, we cloned the Lactococcus lactis gene encoding phospho-beta-galactosidase in L. paramesenteroides. Expression of this heterologous gene in L. paramesenteroides under control of Lactococcus lactis expression signals was evident from the presence, in transformants, of phospho-beta-galactosidase activity and a specific phospho-beta-galactosidase protein band on Western blots (immunoblots). In addition, we transformed a lactose-deficient derivative of L. paramesenteroides with a plasmid carrying a Lactococcus lactis-Escherichia coli lacZ gene fusion. The resulting transformants synthesized high levels of beta-galactosidase, indicating the efficiency of heterologous gene expression signals in L. paramesenteroides.

Blotting, Western↗

2'-5'-oligoadenylate synthetase gene expression in normal and murine sarcoma virus-transformed NIH 3T3 cells.

Mouse fibroblasts transformed by murine sarcoma virus (MSV) are highly sensitive to the antiproliferative effect of interferon (IFN) (M. Bakhanashvili, D. H. Wreschner, and S. Salzberg, Cancer Res. 43:1289-1294, 1983). To elucidate the mechanism leading to this IFN sensitivity, the expression of the 2'-5'-oligoadenylate synthetase (2-5A synthetase) gene, the presence of the 2-5A synthetase protein, and the level of its enzymatic activity were determined in IFN-treated and untreated cultures. NIH 3T3 mouse fibroblasts were compared with two different NIH 3T3 clones transformed by MSV. Cultures were treated with 300 IU of beta IFN (IFN-beta) per ml for 16 to 24 h. While no detectable 2-5A synthetase-derived transcripts were seen in untreated NIH 3T3 cells, two size classes of RNA transcripts, i.e., 1.7 and 4.2 kilobases, were detected in IFN-treated cultures. Surprisingly, a similar amount of transcripts were present in untreated transformed cells. However, following IFN treatment, an eightfold increase in the level of RNA was readily detected in these cells, with no change in the size classes. Similar results were obtained with the 2-5A synthetase protein, for which three size classes of 42, 71, and 102 kilodaltons were demonstrated by immunoblotting, and with the enzymatic activity, for which again, the highest level was seen in IFN-treated MSV-transformed cultures. The basal level of 2-5A synthetase gene expression in the transformed cells has biological significance since these cells were more resistant to mengovirus infection than NIH 3T3 mouse fibroblasts. Medium collected from transformed cultures failed to induce 2-5A synthetase activity in NIH 3T3 cells. Furthermore, antibodies directed against mouse IFN-beta failed to inhibit 2-5A synthetase activity detected in transformed cultures. These results suggest that at least IFN-beta secretion is not involved in the elevated level of 2-5A synthetase gene expression in these cells.

2',5'-Oligoadenylate Synthetase↗

Glucose metabolism and larger molecule removal in long-term high efficiency hemofiltration.

High efficiency hemofiltration (HF) was carried out for 9 months in 6 patients on hemodialysis (HD). Comparative studies on carbohydrate metabolism and 500- to 1,500-dalton serum solute concentrations were performed during HD and HF. After 5 months of HF, intravenous glucose tolerance tests showed an improved peripheral glucose utilization, with unchanged insulin secretion. Larger solute concentrations, measured by gel chromatography, simultaneously dropped in serum suggesting that HF removes some toxic substances that inhibit peripheral insulin action. After 9 months of HF, an unexpected increase in the larger solute concentration coincided with an impaired glucose tolerance, stressing the probable toxic rule of such solutes. Changes in other glucoregulatory hormone secretions were not unequivocal. On account of the later increase in 500- to 1,500-dalton solute concentrations, HF failed to prevent larger toxic molecules accumulating in uremic sera.

Anuria↗

Pre-postdilution haemofiltration.

Postdilution haemofiltration is a treatment indicated for patients who are unable to tolerate standard dialysis. However, its duration is uncomfortably prolonged in cases of low blood flow from the vascular access. In a predilution mode greater clearance rates can be reached even if a greater amount of sterile and pyrogen-free solutions are required. Although an in-line solution-producing system lowers the cost, theoretical calculations indicate that a predilution mode is not advantageous, in terms of the amount of infused solutions, if urea clearances greater than 150 ml/min at 300 ml/min blood flow are required. Theoretically, the combination of predilution and postdilution is the best system to utilise relatively small amounts of sterile solution in order to enhance treatment efficiency at low blood flow. However, the predilution mode may ultimately be preferable because of easier utilisation of the hardware. The results of clinical application of pre-postdilution haemofiltration indicate that at a 400 ml/min blood flow, a urea clearance of more than 220 ml/min can be obtained, and the duration of treatment can be reduced to only 3 h in the majority of patients.

Anuria↗

Immature optic nerve glia of rat do not promote axonal regeneration when transplanted into a peripheral nerve.

The ability of immature central nervous system (CNS) glia to promote axonal regeneration was studied by grafting segments of embryonic and neonatal rat optic nerves into the sciatic nerves of adult rats. Unexpectedly, very few axons regenerated through these grafts. The majority of the axons bypassed the grafts and were associated with Schwann cells. These results were similar to those obtained with grafts of adult rat optic nerves. The failure of immature CNS glia to promote axonal regeneration under these conditions suggests that they may be less effective than Schwann cells in promoting the regeneration and growth of axons.

Aging↗

Laminin and heparan sulphate proteoglycan in the lesioned adult mammalian central nervous system and their possible relationship to axonal sprouting.

Several in vitro studies indicate that the extracellular matrix (ECM) glycoprotein laminin can promote neurite outgrowth from CNS (central nervous system) neurons. Laminin has been detected immunohistochemically in astrocytes in the embryonic but not the uninjured adult mammalian CNS. In the injured adult CNS, it is found in some reactive astrocytes located near the site of CNS lesions. In the present study, we have attempted to examine the relationship between these laminin+ astrocytes and the axonal sprouting that occurs after CNS injuries. This was studied in the intracranially transected adult rat optic nerve which consists of a cranial segment devoid of all retinal ganglion cell axons, and a retinal segment attached to the retina which contains some viable axons that undergo sprouting. Laminin+ reactive astrocytes were found in the cranial segment, but not in the retinal segment. In addition, the cut ends of the retinal and cranial segments were capped by an intensely laminin+, glial fibrillary acidic protein negative (GFAP) region. Axonal sprouts from the transected retinal ganglion cell axons, identified by anterogradely transported rhodamine isothiocynate (RITC), were confined to laminin-, GFAP+ regions of the retinal segment. These results suggest that injury-induced axonal sprouting in the adult mammalian CNS in vivo may be promoted by molecules other than laminin, that may be associated with astrocytes. The presence of heparan sulphate proteoglycan HSP G was also examined in the transected optic nerve because the neurite outgrowth promoting factors found in conditioned media derived from several cell types in vitro have been shown to consist of a complex of laminin and heparan sulphate proteoglycan. No significant changes in heparan sulphate proteoglycan-like immunoreactivity was observed after transection. The presence of laminin and HSPG were also examined in the lesioned adult rat cerebral cortex.

Animals↗

Neurite outgrowth from mammalian CNS neurons on astrocytes in vitro may not be mediated primarily by laminin.

A number of in vitro studies suggest that certain components of the extracellular matrix (ECM), such as the glycoprotein laminin, can promote neurite outgrowth. In the present study the presence of laminin and heparan sulphate proteoglycan, another ECM molecule, on CNS glial (astrocytes) and non-glial (leptomeningeal) cells in vitro was examined. In addition, the ability of these cells and laminin-coated tissue culture substrates to promote neurite outgrowth from developing mammalian cerebellar cortical neurons was also assayed. Leptomeningeal cells were found to be labelled much more intensely with antibodies against laminin and heparin sulphate proteoglycan than were astrocytes. However, the proportion of neurons extending neurites was fourfold greater on astrocyte monolayers than on leptomeninges, and twofold greater than that on laminin. In addition, the length of the neurites growing on astrocyte monolayers was three- to fourfold greater than that on laminin or leptomeninges. Neurite outgrowth on leptomeninges and laminin could not be enhanced by culturing in astrocyte conditioned medium. The ability of various antibodies to block neurite outgrowth on these monolayers was also studied. These results suggest that the robust neurite outgrowth from mammalian CNS neurons plated on astrocytes may be mediated via an astrocyte surface molecule distinct from laminin.

Animals↗