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M Schwartz

Publications and source records attributed to M Schwartz.

At least 631 records · Page 35Linked to original sources

Alterations in mRNA translation products associated with regenerative responses in the retina.

Translation products of mRNA from retinas of goldfish optic nerve (representing a regenerative CNS) and adult rabbit optic nerve (representing a nonregenerative CNS which can be induced to express regenerative characteristics) were examined by one- and two-dimensional gel electrophoresis. Translation products from retinas of the regenerating goldfish optic nerve included polypeptides barely detectable in the translation products of mRNA derived from retinas of uninjured controls. Some of these polypeptides, of apparent molecular weights 24-28, 43-49, 60, and 65 kilodaltons can be considered as growth-associated polypeptides described in other regenerative and developing systems. The induction of regeneration-associated characteristics in the injured adult rabbit optic nerve, "implanted" with diffusible substances from nonneuronal cells of regenerative or growing nerve, is reflected by changes in the mRNA translation products of the retina. Among such translation products are those of the following molecular weights: 16-18, 28, 32-35, 43-47, and 56-60 kilodaltons, and some higher-molecular-weight species.

Animals↗

Essential and nonessential sequences in malPp, a positively controlled promoter in Escherichia coli.

A plasmid bearing the malPp promoter was digested with Bal31 to obtain a set of deletions with closely spaced endpoints in the upstream region of this promoter. Some of these deletions were sequenced, and their effect on malPQ expression was determined after having transferred them onto the chromosome. We found that a site which binds the cyclic AMP receptor protein in vitro and which is centered at position -93 with respect to the site of transcription initiation could be deleted without affecting malPQ expression. In contrast, the activity of the malPp promoter decreased abruptly when the deletions reached position -72. The downstream region of the promoter was analyzed by using a technique of "sequence replacement" which involved the selection of Mal+ pseudorevertants from strains which carried small deletions in the -25 region. The pseudorevertants, which expressed the malPQ operon in a manner indistinguishable from wild type, had grossly different sequences downstream from position -38, except for a few positions, some of which must be important for promoter function. By combining all presently available information, it is suggested that the malPp promoter contains three binding sites for its activator, the product of gene malT. These sites are defined by three quasi-identical hexanucleotides present in one orientation around position -37 and twice in the other orientation around positions -60 and -73.

Base Sequence↗

Characterization and expression of the structural gene for pullulanase, a maltose-inducible secreted protein of Klebsiella pneumoniae.

Some strains of Klebsiella pneumonia secrete pullulanase, a debranching enzyme which produces linear molecules (maltodextrins, amylose) from amylopectin and glycogen. pulA, the structural gene for pullulanase, was introduced into Escherichia coli, either on a multiple-copy-number plasmid or as a single copy in the chromosome. When in E. coli, pulA was controlled by malT, the positive regulatory gene of the maltose regulon. Indeed, pulA expression was undetectable in a malT-negative mutant and constitutive in a malTc strain. Furthermore, the plasmid carrying pulA titrated the MalT protein. When produced in E. coli, pullulanase was not localized in the same way as in K. pneumoniae. In the latter case it was first exported to the outer membrane, with which it remained loosely associated, and was then released into the growth medium. In E. coli the enzyme was distributed both in the inner and the outer membranes and was never released into the growth medium.

Cell Membrane↗

Electrogenicity of the Na-K-ATPase pump in bullfrog cornea epithelium.

The effect of changing the K concentration in the aqueous solution was studied in the frog cornea. In general, when the K concentration was increased from 4 to 20 or 79 mM, the transepithelial PD and resistance decreased. If K was decreased from 79 to 4, 20 to 4, or 4 to 0 mM, or any other combination, the PD and resistance increased. These are normal PD responses. If after a K-free period of more than 5-10 min the K was increased to 4 mM, the PD increased, an anomalous PD response. If K was increased from 0 to 20 mM, there was an initial PD increase (anomalous response) followed by a PD decrease (normal response). If K was increased from 0 to 79 mM the PD decreased, normal response only. The resistance decreased with every increase in K concentration. Anomalous responses were abolished in Na-free solutions and in the presence of both 10(-3) M ouabain and 10(-4) M vanadate in the aqueous solution. We interpret the results on the basis of two pathways, a simple K-conductive pathway and an electrogenic Na-K-ATPase pump pathway with more Na's than K's per cycle. The normal or anomalous PD responses to changes in aqueous K concentration depend on the relative resistance of the two pathways.

Animals↗

Amniotic fluid and advances in prenatal diagnosis.

Maternal blood, amniotic fluid, and chorionic villus samples may be evaluated in the prenatal detection of neural tube defects, cytogenetic disorders, and inborn errors of metabolism. Laboratory tests for these abnormalities usually involve very specialized methods. They should be accompanied by equally rigorous clinical follow-up methods and expert counseling.

Adult↗

Head injuries.

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Brain Injuries↗

Target dependent and independent stages in regeneration.

Goldfish retinae deprived of their target by tectal ablation showed characteristics of regeneration, such as sprouting activity in culture and changes in protein synthesis. These features resemble changes occurring in regenerating retinae of optic nerves which were subjected only to crush injury. Retinae of tectal ablated nerves maintained their ability to sprout in culture longer than retinae of crushed-injured optic nerves. This may indicate a possible absence of the machinery which regulates growth termination as reflected in this case, by the enduring growth activity in culture. Among the changes in protein synthesis which occurred in retinae following either tectal ablation or crush injury, the most pronounced were in tubulin and in some other polypeptides of the following molecular weights: 46-49, 65 and 74 kdalton. Results are discussed with respect to the possible role of the target in initiating regeneration upon disconnection of the nerve, and in terminating growth upon reconnection.

Animals↗

Molecular events associated with increased regenerative capacity of the goldfish retinal ganglion cells following X-irradiation: decreased level of axonal growth inhibitors.

In our previous work we established conditions to study the contribution of non-neuronal cells to the process of goldfish optic nerve regeneration. This issue has been studied successfully by adapting the use of X-irradiation to manipulate division of non-neuronal cells associated with the injured nerve. The regenerative capacity of the goldfish retinal ganglion cells was determined subsequent to the X-ray treatment. In this work we present an analysis of the molecular events associated with regeneration and enhanced regenerative capacity which follows X-irradiation. Under normal conditions the non-neuronal cells surrounding an intact nerve released axonal growth inhibitors, the level of which was only slightly sensitive to X-irradiation. In contrast, regenerating nerves showed a marked decrease in substances having an inhibitory effect on sprouting in vitro. Moreover, their level was significantly reduced following X-irradiation which was accompanied by increased accumulation of fibrous collagen adjacent to astrocytes. The alteration in the reciprocal relationship between the axon and the surrounding non-neuronal cells was also manifested by changes in the profile of labeled proteins released by the non-neuronal cells. The results of this work, therefore, indicate the sensitivity of retinal ganglion cells to the environment provided by the cells surrounding the regenerating fibers and thus may open a new perspective relating to the capacity of neurons to regenerate.

Animals↗

Effect of reduced pH in absence of HCO3(-) on anomalous and normal potential responses in bullfrog antrum.

Effect of changing [K+], [Na+] and [Cl-] in nutrient solution on potential difference (PD) and resistance was studied in bullfrog antrum with and without nutrient HCO3(-) but with 95% O2/5% CO2 in both cases. In both cases, changing from 4 to 40 mM K+ gave about the same initial PD maximum (anomalous response) which was followed by a decrease below control level. Latter effect was much less with zero than with 25 mM HCO3(-). Changing from 102 to 8 mM Na+ gave initial normal PD response about the same in both cases. However, 10 min later the change in PD with zero HCO3(-) was insignificant but with 25 mM HCO3(-) the PD decreased (anomalous response of electrogenic NaCl symport). PD maxima due to K+ and Na+ were largely related to (Na+ + K+)-ATPase pump. Changes in nutrient Cl- from 81 to 8.1 mM gave only a decrease in PD (normal response). Initial PD increases are explained by relative increases in resistance of simple conductance pathways and of parallel pathways of (Na+ + K+)-ATPase pump and Na+/Cl- symport. Removal of HCO3(-) and concurrent reduction of pH modify resistance of these pathways.

Animals↗

Topology of phage lambda receptor protein. Mapping targets of proteolytic cleavage in relation to binding sites for phage or monoclonal antibodies.

Phage lambda receptor protein of Escherichia coli (LamB protein or maltoporin ) was purified in a mild detergent and subjected to prolonged proteolysis by either trypsin or subtilisin. Cleavage occurred at a limited number of sites without affecting the trimeric structure of the protein. Fragments could be dissociated only by heating in sodium dodecyl sulfate to 100 degrees C. The positions of purified fragments were determined with respect to the uncleaved 421-residue polypeptide by chemical analyses. The regions containing target sites were mapped around residues 159, 203, 245, and 370. Based on kinetics of appearance of the different peptides, early cleavage events occurred at sites near residues 159, 203, and 245 and could be distinguished from late events around residue 370. Information regarding the topological orientation of the cleavage sites could be obtained from the effect of in vitro proteolysis on the ability of the protein to bind phage lambda or monoclonal antibodies. Loss of phage lambda neutralizing activity coincided with early cleavage events, whereas loss of antigenic determinants, known to be exposed at the cell surface, appeared late. Cleavage regions are thus likely to be exposed at the cell surface, a conclusion compatible with the location of mutations affecting the interaction of LamB protein with phage in vivo.

Amino Acid Sequence↗

Potential difference responses due to K+, Na+ and Cl- changes in bullfrog antrum with and without HCO3-.

The effect of changing [K+], [Na+] and [Cl-] in nutrient solution was studied in bullfrog antrum with and without HCO3- in nutrient. In 25 mM HCO3- (95% O2/5% CO2) and in zero HCO3- (100% O2), nutrient pH was maintained at 7.3. Changing from 4 to 40 mM K+ or from 81 to 8.1 mM Cl- gave a decrease 10 min later in transmucosal PD (nutrient became more negative)--a normal response. These responses were less in zero than in 25 mM HCO3-. A decrease from 102 to 8 mM Na+ decreased PD (anomalous response of electrogenic NaCl symport). This effect was attenuated or eliminated in zero HCO3-. In contrast, change from 4 to 40 mM K+ gave initial anomalous PD response and change from 102 to 8 mM Na+, initial normal PD response with either zero or 25 mM HCO3-. Both responses were associated with (Na+ + K+)-ATPase pump and were greater in zero than in 25 mM HCO3-. Initial PD increases in zero HCO3- are explained as due to increase in the resistance of passive conductance and/or NaCl symport pathways. Thus, removal of HCO3- modifies conductance pathways of nutrient membrane.

Animals↗

Early regenerative responses induced by monoclonal antibodies directed against a surface glycoprotein of goldfish retinal ganglion cells.

Monoclonal antibodies directed primarily against antigenic determinants associated with the goldfish optic nerve were prepared and characterized. One selected clone 23-4-C(IgG2a), detected antigenic determinants of glycoprotein nature with an apparent mol. wt. of 140 000. Following injury the level of these molecules increased with a peak at 5-7 days after the lesion (2- to 3-fold higher than the basal level). The results strongly suggest that the increase derives, at least partially, from a real increment in the level of these molecules in the retinal ganglion cells rather than from changes in accessibility. Immunofluorescence studies indicate that the retinal ganglion cells carry the antigenicity. Intraocular injection of the monoclonal antibodies, concomitantly with crush injury, resulted in an earlier and higher regenerative response, reflected by sprouting capacity, protein synthesis and accumulation of radiolabeled material in the tectum contralateral to the side of injury. This may indicate that the antibodies directly activate retinal cells via interaction with surface molecules. Alternatively, the antibodies may be directed against surface molecules which are associated with axonal growth inhibitors, and may therefore mask these surface antigens from further interaction with their native substrate.

Animals↗

Colicin E2 release: lysis, leakage or secretion? Possible role of a phospholipase.

Results presented here and by others indicate that the release of colicins from producing cells can be uncoupled from the decline in culture turbidity which usually occurs within 2-3 h after the induction of colicin synthesis. This excludes lysis as a necessary event in colicin release. Conversely, the failure to dissociate colicin release from the normally simultaneous release of a specific subset of soluble proteins argues against the idea of a specific colicin secretion system sensu-stricto. Rather, colicin release appears to be a consequence of semi-specific leakage resulting from an alteration of the permeability properties of the cell envelope. This alteration is caused by the 'lysis protein' known to be encoded by most multiple copy number Col plasmids. The finding that the expression of the lysis gene of plasmid ColE2 renders the cells exquisitely sensitive to lysozyme demonstrates that the permeability of the outer membrane must indeed be altered. Evidence is presented that this alteration could be due at least in part to the activation of the detergent-resistant phospholipase A (pldA product). Lysophosphatidylethanolamine, a product of the action of phospholipase on phosphatidylethanolamine, is a membrane perturbant which could alter the permeability properties of the envelope and allow some proteins such as colicin to leak out of the cell.

Cell Survival↗

Antibodies to gangliosides inhibit goldfish optic nerve regeneration in vivo.

Intraocular injection of antiserum to mixed ganglioside or to GM1 inhibited the regeneration of goldfish optic axons following an optic nerve crush. For example, injections of antiserum on 5 consecutive days beginning the day before the crush resulted in a decrease of about 40% in the axonal outgrowth distance measured at 10 days after the crush. The inhibition was observed even when the treatment was begun a few days after the lesion, and greater degrees of inhibition were observed when the treatment was given later in regeneration. This indicates that the antiganglioside serum interfered with axonal elongation more than with the initial sprout formation. The antiganglioside treatment did not impair the enlargement of the cell bodies and nucleoli that accompanies regeneration, nor did it affect fast axonal transport of protein, glycoprotein, or glycolipid in regenerating nerves. Thus inhibition of outgrowth by antiganglioside treatment was not mediated by a gross change in the metabolism of the regenerating neurons. Treatment of normal neurons with the antiserum produced a 20-30% increase in the amount of 3H-glucosamine-labeled glycoproteins and glycolipids conveyed by fast axonal transport. These results suggest that membrane gangliosides may normally influence the supply of axonally transported glycosylated macromolecules. However, the effect of antiganglioside on axonal transport of glycosylated molecules and on axonal outgrowth are not necessarily related to each other.

Animals↗

Effects of luminance and flicker on ocular dominance shift in kitten visual cortex.

We raised monocularly deprived kittens in visual environments with low level illumination that was either steady or flickering. With steady scotopic luminance ocular dominance shifted as it does in normal photopic lighting. In flickering light with an average frequency of 2 Hz there was virtually no ocular dominance shift, while in flickering light averaging 0.1 Hz there was a significant shift. Recordings from the 2 Hz flicker-reared were similar to the dark-reared recordings. The flickering illumination was produced in one case by a high contrast-low brightness TV near the cage, and in another case, by a low voltage incandescent bulb driven by a pseudo-random sequence generator. This circuit delivered either a maximum ON time of 1.7 s or a maximum of 40 s, for the 2 Hz and 0.1 Hz respectively. Both the TV and flickering bulb produced average illumination comparable to the dim (0.01 cd/m2) steady scotopic illumination. We conclude that dim flickering light is not a sufficient stimulus for promoting ocular dominance shift in kittens in the critical period unless the flicker rate approaches 0.1 Hz. Furthermore results from the TV rearing suggest that flicker may be capable of preventing an ocular dominance shift expected from a concurrent steady low light level background.

Animals↗

Hexosaminidase: a biochemical marker for necrotizing enterocolitis in the preterm infant.

To evaluate hexosaminidase as a biochemical marker for the early detection of necrotizing enterocolitis in preterm infants, 33 preterm infants without necrotizing enterocolitis and 18 preterm infants with necrotizing enterocolitis had hexosaminidase activity measured during the course of their hospitalization. Although hexosaminidase activity could not identify those preterm infants with necrotizing enterocolitis who had impending perforation, the data strongly suggest that measurements of serum hexosaminidase activity may provide an early biochemical indication of the presence of necrotizing enterocolitis in the preterm infant. Hopefully, early detection of necrotizing enterocolitis in these infants will reduce the morbidity and mortality of this disease.

Clinical Enzyme Tests↗

Regenerative capacity of the goldfish visual system is affected by antibodies specific to gangliosides injected intraocularly.

An association between gangliosides and neuronal regeneration in goldfish is demonstrated in the present study. A single intraocular injection of affinity purified anti-GM1 antibodies administered simultaneously with crush injury of the optic nerve, inhibits the regenerating process as expressed by two parameters: protein synthesis in the retina and in vitro sprouting ability from the retina. The retinal level of several gangliosides (such as GD3, GD1a, GD1b and GT1b) is enhanced during regeneration. Although GM1 appears to be a minor retinal ganglioside, antibodies to GM1 exert a marked effect on retinal regenerative process. It is assumed that such antibodies could interact with more abundant retinal gangliosides such as GD1b which shows enhanced biosynthesis during regeneration and which shares a similar disaccharide terminal residue with GM1.

Animals↗

Adult erotosexual status and fetal hormonal masculinization and demasculinization: 46,XX congenital virilizing adrenal hyperplasia and 46,XY androgen-insensitivity syndrome compared.

Among 30 young women with a history of the treated adrenogenital syndrome (CVAH), 11 (37%) rated themselves as bisexual or homosexual. Among a control group consisting of 15 women with the 46,XY androgen-insensitivity syndrome (AIS) plus 12 with the Rokitansky syndrome (MRKS), the corresponding figure was 2 (7%), both bisexual. Chi-square was significant beyond the 0.01 level. In Kinsey's 1953 sample 15% of women experienced homoerotic arousal imagery by age 20, and 10% had had homoerotic partner contact. The most likely hypothesis to explain the CVAH findings is that of a prenatal and/or neonatal masculinizing effect on sexual dimorphism of the brain in interaction with other developmental variables.

Adrenal Hyperplasia, Congenital↗