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Biomedical subjects

E Sanders

Publications and source records attributed to E Sanders.

At least 37 records · Page 2Linked to original sources

The Mycobacterium tuberculosis 38-kDa antigen: overproduction in Escherichia coli, purification and characterization.

The 38-kDa protein (Ag38) of the Gram+ bacterium, Mycobacterium tuberculosis H37Rv, is an immunodominant antigen of potential utility for diagnosis and vaccine development. Assessment of this potential requires large amounts of the purified protein that would be difficult, if not impossible, to obtain from M. tuberculosis itself. The gene coding for Ag38 had been previously cloned and in the present study was expressed as an unfused protein in Escherichia coli under the control of strong transcriptional (bacteriophage lambda pLpR) and translational (atpE) signals. Fermentation of the recombinant E. coli K-12 strain CAG629[pMS9-2], which is deficient in Lon protease and the heat-shock response, produced recombinant Ag38 (reAg38) at high levels (about 10% of total cellular protein). The reAg38, which accumulated as inclusion bodies, was completely solubilized in 6 M guanidine.HCl, refolded and purified to apparent homogeneity. The product showed the expected amino acid composition and M(r), and had similar reactivities as the native protein with three different mAb. Polyclonal antibodies raised against reAg38 reacted strongly with the native antigen in enzyme-linked immunosorbent assay. These results demonstrate that reAg38, which cannot be distinguished antigenically from the native protein of M. tuberculosis, can be prepared in quantity from E. coli.

Amino Acid Sequence↗

Subacute idiopathic demyelinating polyradiculoneuropathy.

Seven cases of subacute idiopathic demyelinating polyradiculoneuropathy had a monophasic illness characterized by progressive weakness of all four limbs that evolved during 4 to 8 weeks. Neurophysiological investigations implied demyelination in all seven cases. In two patients, sural nerve biopsy specimens that were taken showed macrophage-associated demyelination. All patients made substantial or complete recoveries with oral prednisolone (four cases) or without treatment (three cases). None of the patients required ventilation or had autonomic complications. These cases provide a link between the acute idiopathic demyelinating form of Guillain-Barré syndrome and chronic idiopathic demyelinating polyradiculoneuropathy.

Adult↗

High cell density fermentation of recombinant Escherichia coli with computer-controlled optimal growth rate.

In recent years recombinant DNA technology has enabled us to produce various proteins of therapeutic importance with microorganisms. As an appropriate host organism, E. coli plays a dominant role. Yields of E. coli dry cell mass in shaker flask culture range from 1-2 g/L, whereas in fermentors up to 10 g dry cells/L can be achieved. ZIMET and GBF have developed a high cell density fermentation process that produces E. coli (on a glucose/mineral salt medium) up to more than 100 g dry cells/L in a special fed-batch mode. This cultivation strategy prevents oxygen limitation and hence the accumulation of acetate and other metabolic byproducts. The specific growth rate can be adjusted so that product formation reaches its optimum value. An example of the production of alpha1-interferon is presented. The high cell density fermentations were realized in 30- and 450-L Chemap fermentors (ZIMET) and in a three-stage bioreactor scale-up system (72, 300, and 1,500 L) developed in cooperation with GBF and B. Braun Melsungen AG. Multiloop controllers were used to control the process variables.

Biotechnology↗

Bone-alkaline phosphatase as indicator of bone formation.

Bone-alkaline phosphatase was determined in patients at risk of osteoporosis due to treatment with oral corticosteroids, and in patients at risk of increased bone synthesis because of treatment with cyclosporin. Both a significant decrease of bone-alkaline phosphatase during corticosteroid treatment, and a significant increase of bone-alkaline phosphatase during cyclosporin treatment could be demonstrated. It is concluded that bone-alkaline phosphatase is a useful parameter for monitoring changes in bone formation.

Adult↗

Isomyosin expression in developing chicken atria: a marker for the development of conductive tissue?

Isomyosin expression patterns in embryonic chicken atria during the first two weeks of development were analyzed immunohistochemically. In the 3-days embryonic chicken heart (HH19-20), strong coexpression of both isomyosins can be found as band-like zones at the lateral sides of the sinoatrial junction. The zones converge on the bottom of the atrium and continue as a band around the atrioventricular canal. In the 5-days heart (HH27-28) the coexpression area encompasses the entire sinoatrial junction and extends into parts of the sinus venosus and into the dorsocaudal atrial wall. In the 7-days heart (HH 32-33) the relative extension of coexpression areas reaches its maximum. Coexpression is also found in a ring-like band in the ventral (bottom) wall of the atria peripheral to the ring-like band in the atrioventricular junction. The latter band has now become continuous with the coexpression area in the bottom of the interatrial septum. Caudally coexpression extends behind the atrioventricular cushions towards the interventricular septum and cranially coexpression of the atrioventricular junction has become continuous with that of the ring around the outflow tract (cf Sanders et al. 1986). In the second week of incubation a decrease of coexpression is observed. The isomyosin expression pattern described in this study has put forward additional arguments that the conductive tissue originates from areas that continue to express both isomyosins relatively late in development.

Animals↗

The local expression of adult chicken heart myosins during development. II. Ventricular conducting tissue.

The development of the ventricular conducting tissue of the embryonic chicken heart has been studied using a previous finding that morphologically recognizable atrial conducting tissue coexpresses the atrial and the ventricular myosin isoforms. It is found that, by these criteria, at 9 days part of the ventricular conduction system consists of a myocardial ring located around the infundibula of the aorta and truncus pulmonalis. Part of this ring is formed by the retro-aortic root branch. The ring continues via the septal branch into the atrioventricular bundle and its branches, that all express both myosin isoforms. The retro-aortic root branch could be traced back as a part of the myocardial wall of the truncus arteriosus at the 4 days embryonic stage. At the 16th day of development, the septal branch, atrioventricular bundle and left and right bundle branches no longer express the atrial isomyosin, but two bundles originating from the septal branch still express both isomyosins, one being the retro-aortic root branch, the other being only immunologically recognizable and directed to the ventral side of the truncus pulmonalis; this latter we call the pulmonary root branch. Both bundles are remnants of the myocardial ring.

Animals↗

The conducting tissue in the adult chicken atria. A histological and immunohistochemical analysis.

A three-dimensional reconstruction from serial sections of adult chicken heart was made to verify whether Purkinje cells, that can be recognized by a number of well-known histological criteria, form specialized tracts in the adult chicken atria. This reconstruction revealed a loosely arranged network of Purkinje cells connecting the two atria. This network has not been described before. No tracts could be detected between the sinoatrial and the atrioventricular nodes. These atrial Purkinje cells express the atrial and ventricular myosin isoform, as determined by the use of monoclonal antibodies that were prepared against atrial and ventricular myosin isoform, respectively. Some atrial myocytes that are topographically closely related to the Purkinje cells and that cannot be distinguished from the surrounding myocytes with conventional histological criteria, express, apart from the atrial myosin isoform, also the ventricular myosin isoform. The similar expression pattern of these two cell types and their close topographical relationship suggest the presence of a more elaborate system specialized in conduction than the well-known conductive system found with conventional histological techniques.

Animals↗

The local expression of adult chicken heart myosins during development. I. The three days embryonic chicken heart.

Immunofluorescence studies were performed on serial sections of three days embryonic chicken hearts using antibodies specific for adult atrial and ventricular myosin heavy chains respectively. The anti-ventricular myosin serum reacted with the entire myocardium showing a decreasing intensity going from the truncus arteriosus to the atrial part; however, the antiatrial myosin serum reacted weakly with the myocardium of the atrial part. Two other interesting observations were made, i) the anti-atrial myosin serum reacted with non-myocardial cells in the cardiac jelly, ii) both antisera reacted with a thin myocardial layer, extending from the ventral wall of the atrial part via the medio-dorsal wall of the atrio-ventricular canal to the dorsal wall of the ventricular part.

Animals↗

Cortical distribution of prostaglandin and renin in isolated dog glomeruli.

This study was performed (1) to evaluate the ability of renal superficial (SP) and juxtamedullary (JM) isolated glomeruli to synthesize 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha) and prostaglandin E2 (PGE2) when stimulated with different doses of arachidonic acid (AA) and (2) to determine the differential effects that the formation of these prostaglandins may have on renin release. Renal SP and JM glomeruli were isolated separately from dog renal cortex using a passive sieving technique. Glomeruli were superfused within glass chambers with a Krebs Ringer solution and three different concentrations of AA. Effluent concentrations of 6-keto-PGF1 alpha (primary metabolite of PGI2), PGE2, and renin were determined by radioimmunoassay. Synthesis of 6-keto-PGF1 alpha by SP glomeruli increased in a dose-dependent manner in response to the three increasing concentrations of AA. These three doses of AA also evoked proportional increments in the release of renin which were significantly correlated with the increases in 6-keto-PGF1 alpha. In contrast, PGE2 synthesis was maximally stimulated by all doses of AA and was not correlated with renin release. In JM glomeruli, AA evoked the same pattern in the synthesis of 6-keto-PGF1 alpha and PGE2. However, renin release increased significantly only with the perfusion of the highest concentration of AA. These perfusion results show that SP and JM glomeruli have the same ability to synthesize 6-keto-PGF1 alpha and PGE2. However, renin release was significantly higher in SP than in JM glomeruli and correlated with 6-keto-PGF1 alpha synthesis.

6-Ketoprostaglandin F1 alpha↗

Computer-aided emergency telecommunications for the deaf.

Computer-aided telecommunications provide deaf teletypewriter users with 24-hour toll-free access to emergency services. An interface and software link the deaf caller's teletypewriter (TDD) with a microprocessor by reducing and inverting voltage levels between the two devices. This system facilitates rapid transmission of linguistically controlled triage questions to meet the communication needs of deaf patients.

Communication Devices for People with Disabilities↗

Renin release selectively stimulated by prostaglandin I2 in isolated rat glomeruli.

Renal glomeruli were isolated from rat kidneys using a passive mechanical sieving technique. Glomerular microsomal fraction, glomerular homogenate, or intact glomeruli were incubated with [1-14C]arachidonic acid, and the profile of prostaglandin (PG) synthesis was determined by thin-layer chromatography. The three incubation systems produced 15.3, 20.8, and 40.4% 6-keto-PGF1 alpha; 19.1, 23.5, and 15.3 PGF2 alpha; 5.7, 9.1, and 3.9% thromboxane (TX) B2; 36.0, 35.1, and 37.0% PGE2; and 23.9, 11.3, and 3.4% PGD2, respectively. Glomeruli were placed in suspension within glass chambers and superfused with Krebs solution. Superfusion with 1.6 x 10(-4) M arachidonic acid stimulated a significant release of renin from glomeruli, whereas 2.7 x 10(-6) M PGE1, PGE2, PGF2 alpha, TXB2, PGD2, or a stable analog of PGH2 had no effect on renin. When the rapid breakdown of PGI2 was counteracted by either increasing the concentration to 1.7 x 10(-4) M or stabilizing in Krebs at pH 9.4, it stimulated a significant increase in renin release. Reducing the arachidonic acid concentration to 1.6 x 10(-5) M eliminated both renin release and PGI2 synthesis, while increased PGE2 synthesis persisted. Finally, using an inhibitor of PGI2 synthesis, azo analog 1 (2.8 x 10(-6) M), 6-keto-PGF1 alpha produced in response to arachidonic acid was eliminated, as was the concurrent release of renin, but PGE2 synthesis was not affected. These results suggest that the mechanism of direct interaction between renal PG and renin in isolated glomeruli is selectively due to the action of PGI2.

Animals↗

On the pathogenesis of glomerulonephritis: a clinico-pathological study indicating that neutrophils attack and degrade glomerular basement membrane.

Lysosomal proteinase activity was assayed in urines from 157 patients with renal disease. 16 were found to be excreting acid and neutral proteinases. 15 of these 16 had a severe proliferative glomerulonephritis with polymorphonuclear leucocyte (PMN) infiltration of the glomeruli. These same 16 patients excreted excess quantities of glomerular basement membrane (GBM) antigen. Only urines with neutral proteinase activity were capable of degrading GBM in vitro. Clinical recovery was accompanied by disappearance of both proteinases and excess GBM fragments from the urine. Detailed studies on one patient showed that the neutral proteinase activity was, at least in part, due to PMN lysosomal elastase. Calculation shows that in these patients enough PMN neutral proteinase was present in the glomeruli to cause significant GBM damage.

Antigens↗