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Biomedical subjects

M A Goldstein

Publications and source records attributed to M A Goldstein.

At least 37 records · Page 2Linked to original sources
PubMed1993

The hydrophobic repeated domain of the Clostridium cellulovorans cellulose-binding protein (CbpA) has specific interactions with endoglucanases.

We overexpressed one of the hydrophobic repeated domains (HBDs) (110 amino acid residues) of the cellulose-binding protein (CbpA) from Clostridium cellulovorans by making a hybrid protein with the Escherichia coli maltose-binding protein (MalE). The HBD was purified to homogeneity, and interactions between the HBD and endoglucanases were analyzed by a novel interaction Western blotting (immunoblotting) method. The HBD had specific interactions with endoglucanases (EngB and EngD) from C. cellulovorans. These results indicated that the HBD was an endoglucanase binding site of CbpA.

ATP-Binding Cassette Transporters↗
PubMed1993

Targeted developmental overexpression of calmodulin induces proliferative and hypertrophic growth of cardiomyocytes in transgenic mice.

Calmodulin (CaM) levels are developmentally regulated in the mouse heart. During late gestational and early postnatal stages, CaM levels decline several-fold in close temporal association with the declining population of proliferating cardiomyocytes. This correlation suggests that CaM may influence cardiomyocyte cell cycle activity, particularly since CaM is implicated in cell cycle control in several eukaryotic nonmuscle cells. To test this possibility, nucleotides -500 to 77 of the human atrial natriuretic factor gene were linked to a chicken CaM minigene to establish two pedigrees of transgenic mice that express 3- to 5-fold increased levels of CaM in cardiomyocytes. Developmental overexpression of CaM in mouse cardiomyocytes produced a markedly exaggerated cardiac growth response, characterized by the presence of cardiomyocyte hypertrophy in regions demonstrated to overexpress CaM and by cardiomyocyte hyperplasia, apparent at early developmental stages. Early postnatal suppression of fusion gene expression in the cardiac ventricles correlated with regression of the ventricular growth response in transgenic relative to nontransgenic mice between 3 days and 6-10 weeks of age, but was not apparent in the cardiac atria, where levels of CaM remained constitutively elevated until advanced stages. To test the possibility that increased cytosolic Ca2+ buffering contributes to the growth response induced by CaM over-expression, two additional lines of transgenic mice were generated using the same human atrial natriuretic factor promoter to target expression of a CaM mutant (amino acids 75-82 deleted) in cardiomyocytes. This mutant has previously been shown to bind Ca2+ with kinetic properties similar to those of wild-type CaM, but was unable to activate several CaM-dependent target enzymes in vitro. Despite high level expression of the CaM mutant, no growth response was apparent in the hearts of transgenic relative to those of nontransgenic mice, suggesting that increased Ca2+ buffering is unlikely to contribute to the growth response induced by CaM overexpression. Taken together, these findings reveal that cardiomyocyte growth regulation is specifically influenced by CaM concentrations in transgenic mice.

Animals↗
PubMed1992

Primary sequence analysis of Clostridium cellulovorans cellulose binding protein A.

The cbpA gene for the Clostridium cellulovorans cellulose binding protein (CbpA), which is part of the multisubunit cellulase complex, has been cloned and sequenced. When cbpA was expressed in Escherichia coli, proteins capable of binding to crystalline cellulose and of interacting with anti-CbpA were observed. The cbpA gene consists of 5544 base pairs and encodes a protein containing 1848 amino acids with a molecular mass of 189,036 Da. The open reading frame is preceded by a Gram-positive-type ribosome binding site. A signal peptide sequence of 28 amino acids is present at its N terminus. The encoded protein is highly hydrophobic with extremely high levels of threonine and valine residues. There are two types of putative cellulose binding domains of approximately 100 amino acids that are slightly hydrophilic and eight conserved, highly hydrophobic beta-sheet regions of approximately 140 amino acids. These latter hydrophobic regions may be the CbpA domains that interact with the different enzymatic subunits of the cellulase complex.

Amino Acid Sequence↗
PubMed1992

Cardiac morphology after conditions of microgravity during COSMOS 2044.

Light- and electron-microscopic studies were performed on cardiac muscle from rats flown on COS-MOS 2044 and from four control groups. Average cross-sectional area of myofibers was measured by video analysis of the light-microscopic images of papillary and ventricular muscle samples from all animals. This cross-sectional area was significantly decreased in flight rats (P = 0.03) compared with synchronous controls. Additional findings at the electron-microscopic level consistent with this atrophy were obtained by stereological analysis and optical diffraction analysis of papillary muscle samples. Slightly higher mitochondrial volume density values and mitochondria-to-myofibril ratios as well as normal A-band spacings (d1,0) and Z-band spacings of myofibrils were observed in the tail-suspension and flight groups. General morphological features similar to those in ventricular samples from the previous COSMOS 1887 flight were observed.

Animals↗
PubMed1991

Similar features in Z bands of both skeletal and cardiac muscle revealed by image enhancement.

We have shown previously that the small square (ss) and basket weave (bw) states of the Z band lattice in cardiac and skeletal muscle are related to the contractile state of the muscle. We have used two-dimensional image processing techniques on digitized electron micrographs to enhance the structural features of each projected lattice form in cardiac and skeletal muscle. Four different processing techniques were employed to assess the effect of enhancement artifacts on the resulting Z band images. We observed only slight differences between enhanced images of a particular Z band form produced by the four different techniques. Every enhanced image showed an approximate four-fold symmetry independent of muscle type or Z band lattice form. Each enhanced image showed four cross-connecting Z-filaments which appeared to connect each axial filament to the four nearest axial filaments. In bw images from both cardiac and skeletal muscle, axial filaments had a greater apparent diameter and a greater interaxial filament spacing than in the ss images. In both muscle types, the cross-connecting Z-filaments appeared to overlap half-way between axial filaments in the ss images while the bw images showed no such overlap. These structural features are consistent with a dynamic Z band lattice that participates in muscle contraction.

Algorithms↗
PubMed1991

Role of the Z band in the mechanical properties of the heart.

In striated muscle the mechanism of contraction involves the cooperative movement of contractile and elastic components. This review emphasizes a structural approach that describes the cellular and extracellular components with known anatomical, biochemical, and physical properties that make them candidates for these contractile and elastic components. Classical models of contractile and elastic elements and their underlying assumptions are presented. Mechanical properties of cardiac and skeletal muscle are compared and contrasted and then related to ultrastructure. Information from these approaches leads to the conclusion that the Z band is essential for muscle contraction. Our review of Z band structure shows the Z band at the interface where extracellular components meet the cell surface. The Z band is also the interface from cell surface to myofibril, from extra-myofibrillar to myofibril, and finally from sarcomere to sarcomere. Our studies of Z band in defined physiologic states show that this lattice is an integral part of the contractile elements and can function as an elastic component. The Z band is a complex dynamic lattice uniquely suited to play several roles in muscle contraction.

Animals↗
PubMed1991

Hydrogen peroxide pretreatment of perfused canine vessels induces ICAM-1 and CD18-dependent neutrophil adherence.

BACKGROUND: Cytotoxic products of neutrophils (polymorphonuclear leukocytes, PMNs) contribute to ischemia-reperfusion injury of several tissues. Hydrogen peroxide (H2O2), one of the cytotoxic products of PMNs, also promotes the adherence of PMNs to cultured vascular endothelial cells in vitro. The present study was undertaken to determine if H2O2 also augmented adhesion of PMNs to intact vessels perfused ex vivo and to determine if H2O2-induced PMN adherence to intact canine carotid arteries and external jugular veins or to cultured canine venous endothelium is mediated by specific adherence ligands on the neutrophil and/or the endothelium. METHODS AND RESULTS: Vessels were perfused for 20 minutes with oxygenated Krebs-Henseleit bicarbonate buffer with and without H2O2, washed with buffer alone, and then exposed to 111In-labeled isolated PMNs (10(7) cells/vessel) under static conditions for up to 20 minutes before being washed again. Residual radioactivity retained by the washed vessel was counted as an index of PMN retention. The adherence of unlabeled PMNs to cultured endothelial cells was determined by a visual assay method after pretreatment of the endothelium with H2O2 for brief periods followed by washing. Perfusion of vessels with H2O2 produced a transient, concentration-dependent increase in PMN adhesion to both canine carotid arteries and external jugular veins that was two to four times that of control values at 1 mmol/l and declined at higher H2O2 concentrations. Peak retention of PMNs by canine carotid arteries occurred 10 minutes after exposure to 1 mmol/l H2O2 and then rapidly declined to control values; this effect was replicated by a second 20-minute exposure of canine carotid arteries to 1 mmol/l H2O2 60 minutes after the first exposure. Scanning and transmission electron microscopy revealed not only adherence of PMNs to but migration through the vascular endothelium of the carotid artery after H2O2 perfusion. The endothelium was intact in H2O2-treated arteries not exposed to PMNs. H2O2-induced PMN retention was completely inhibited by addition of catalase or the hydroxyl radical scavenger dimethylthiourea to the perfusate by incubation of the PMN with a monoclonal antibody (Mab) against CD18 (R15.7) or by perfusion of the H2O2-treated vessel with CL18/6, a Mab against canine ICAM-1 (intercellular adhesion molecule-1). Similar effects of Mabs on PMN adhesion to H2O2-pretreated cultured endothelium were noted. The retention of PMNs by vessels mechanically denuded of endothelial cells was markedly increased. H2O2 pretreatment of these vessels did not further augment PMN adherence, and no inhibitory effect of R15.7 was noted. Incubation of carotid arteries and PMNs with a specific platelet-activating factor antagonist, WEB2086, completely inhibited the H2O2-induced increased PMN retention by these vessels. CONCLUSIONS: These results indicate that H2O2 in the absence of evidence for permanent endothelial cell injury, can induce a transient, reversible, platelet-activating factor-dependent adherence of PMNs to vessels by mechanisms that depend on an intact endothelium and involve CD18 on the PMN and ICAM-1 on the endothelium.

Animals↗
PubMed1990

Efficacy of transtelephonic electrocardiographic monitoring in pediatric patients.

The results of transtelephonic electrocardiography for transient symptomatic event recording in 61 consecutive pediatric patients (aged 4 months to 21 years) were reviewed. In 13 (21%) of 61 patients, previous arrhythmia diagnoses had been made by electrophysiologic study; monitoring was utilized to evaluate symptoms before or after pharmacological/surgical therapy. Transtelephonic electrocardiography was used to study undocumented symptomatic events consistent with an arrhythmia in the remaining 48 (79%) of 61 patients. Adequate transmissions during symptoms were obtained in 36 (59%) of 61 patients. Goals of monitoring were achieved in all 13 patients with previous diagnoses. Of the 25 of 48 patients without prior diagnoses who provided transmissions during symptoms, 15 had sinus rhythm, while 10 manifested an arrhythmia. Abnormal transmissions were most common in patients with palpitations and never evident in patients with symptoms of chest pain.

Adolescent↗
PubMed1990

Two structural states of the vertebrate Z band.

Ultrastructural analysis of the vertebrate Z band suggests that two reversible states of a single intricate lattice are essential for the contractile process. The two structural states of the Z band lattice (ss and bw) have been described in cross section in skeletal and cardiac muscle in different physiological states. The lattice responds to active tension but resists passive deformation. Changes in Z band form and dimension are correlated with cross-bridge binding. Two-dimensional image processing techniques show enhanced structural features that vary with the observed changes in lattice dimension. All projected images from all lattices show an approximate four-fold symmetry. Each image reveals differences in the appearance of axial filaments which enter from opposite sides of the Z band and cross-connecting filaments of similar curvature which appear to connect each axial filament to four nearest axial filaments. In the ss images, the apparent diameter of cross-cut axial filaments and the Z band interaxial filament spacing are smaller than in bw images. Cross-connecting filaments appear to overlap in the region half-way between axial filaments in ss images. We conclude that the Z band is an essential and dynamic part of the sarcomere, uniquely suited to transmit tension while maintaining dimensions appropriate for cross-bridge interaction.

Animals↗
PubMed1989

Superoxide generation by photomediated redox cycling of anthraquinones.

Anthraquinones (AQs) comprise one important class of secondary metabolites predominantly produced by fungi and higher plants but also produced by a variety of other organisms. Humans orally ingest AQs from environmental sources as well as through direct use as nonprescription laxatives, and some AQ derivatives are used as topically applied antipsoritic agents. Some AQs are mutagenic. We present evidence that aqueous solutions of several AQs in the presence of an appropriate reducing agent and dissolved oxygen generate superoxide when they are illuminated with broad-spectrum light. Redox cycling of AQs could be responsible for some aspects of their toxicity in biological systems.

Anthraquinones↗
PubMed1989

Comparison of types of pulmonary stenosis with the state of the ventricular septum in complete transposition of the great arteries.

The pulmonary valve and left ventricular outflow tract of 214 hearts with d-transposition of the great arteries (d-TGA) were visually inspected (126 of 214 with intact ventricular septum and 88 of 214 with ventricular septal defect [VSD]). Pulmonary stenosis (PS), either valvular, subvalvular, or in combination, was present in 26 cases and was found to be more common in the presence of a VSD (20.5%) than intact ventricular septum (6.3%). PS occurred more commonly in the presence of a supracristal VSD than an infracristal VSD (70%, 7 of 10 vs 15%, 10 of 66). Further, infracristal or supracristal VSDs were associated with different types of pulmonary obstruction. In seven of ten cases with infracristal VSD and PS, the stenosis was caused by an anomaly of an atrioventricular valve. In six of seven cases with supracristal VSD and PS, the latter was caused by membranous encirclement of the left ventricular outflow tract.

Cardiomyopathy, Hypertrophic↗
PubMed1989

The Z-band lattice in skeletal muscle in rigor.

Previous work with tetanized and relaxed muscle has shown a correlation between active tension and the structure of the Z-band. This suggests that there is a correlation between the cross-bridge binding in the A-band and the structure of the Z-band. Using electron microscopy and optical diffraction we have examined this correlation in glycerinated muscle in rigor and in unstimulated intact muscle. We have found that the Z-bands of muscles in rigor always show the basketweave form, while those of the unstimulated muscles always show the small square form. The basketweave form found in rigor muscles is similar in form and dimension to that found in tetanized muscle. Thus it appears that the small square form of the Z-band is found in physiological states with little cross-bridge binding and the basketweave form is found in states with a high degree of cross-bridge binding.

Animals↗
PubMed1989

Two structural states of Z-bands in cardiac muscle.

We have compared the form and dimensions of the Z-band lattice in rat papillary muscle fixed at rest with and without ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) using electron microscopy and optical diffraction. In unstimulated muscle, the Z-band lattice form called basket weave predominated, and the Z-spacing (defined as the repeat distance of a tetragonal array of cross-cut thin filaments from the same sarcomere) was 23.93 +/- 0.37 nm. Muscles exposed to EGTA exhibited the small square-lattice form, and the Z-spacing was 20.50 +/- 0.19 nm. The Z-spacings in the two lattice forms were similar in cardiac and skeletal muscles such that the decrease in Z-spacing in the transition from basket weave to small square in this study was similar to the increase in Z-spacing previously demonstrated in skeletal muscle in the transition from small square to basket weave. The Z-lattice form and dimensions in unstimulated cardiac muscle resembled those in tetanized skeletal muscle. These findings are consistent with the higher resting tension in cardiac muscle and suggest that Ca2+ may be important for the maintenance of the expanded Z-lattice form.

Actin Cytoskeleton↗
PubMed1988

Structural states in the Z band of skeletal muscle correlate with states of active and passive tension.

In skeletal muscle Z bands, the ends of the thin contractile filaments interdigitate in a tetragonal array of axial filaments held together by periodically cross-connecting Z filaments. Changes in these two sets of filaments are responsible for two distinct structural states observed in cross section, the small-square and basketweave forms. We have examined Z bands and A bands in relaxed, tetanized, stretched, and stretched and tetanized rat soleus muscles by electron microscopy and optical diffraction. In relaxed muscle, the A-band spacing decreases with increasing load and sarcomere length, but the Z lattice remains in the small-square form and the Z spacing changes only slightly. In tetanized muscle at sarcomere lengths up to 2.7 micron, the Z lattice assumes the basketweave form and the Z spacing is increased. The increased Z spacing is not the result of sarcomere shortening. Further, passive tension is not sufficient to cause this change in the Z lattice; active tension is necessary.

Animals↗
PubMed1987

DNase I interactions with filaments of skeletal muscles.

DNase I-actin interactions were studied by electron microscopy and SDS-polyacrylamide gel electrophoresis. Electron micrographs of glycerinated avian pectoralis major muscle fibres treated with 1 mg ml-1 DNase I demonstrated that the Z-lattice was destroyed. The axial filaments of the Z-lattice were still present but were thinner and less ordered than those of control fibres. The small diameter cross-connecting filaments of the Z-lattice were not present in DNase I treated muscle fibres. Treatment with DNase I had the same effect on fast avian skeletal muscles and fast and slow rat skeletal muscles suggesting that the effect was not muscle type or species dependent. The effect of DNase I could be abolished by lowering the DNase I concentration (0.1 mg ml-1 or less). Preincubation of DNase I with purified skeletal muscle actin also abolished the effect of the DNase I. Analysis of the extracts obtained during DNase I treatment by gel electrophoresis demonstrated that substantial quantities of DNase I did not remain associated with the myofibrils. Two proteins, one with an apparent molecular weight of 43 kDa which reacted with an actin antibody and another with an apparent molecular weight of 95 kDa which did not react with an alpha-actinin antibody, were observed in the DNase I extracts. These data suggest that DNase I interacts with protein(s) in the Z-lattice and this interaction causes the subsequent release of actin and other Z-band proteins.

Actin Cytoskeleton↗
PubMed1987

Z band dynamics as a function of sarcomere length and the contractile state of muscle.

The Z band in skeletal muscle has two distinct structural states--a relaxed (small square or ss) form and a maximally activated (basket weave or bw) form. We have examined by electron microscopy and optical diffraction Z lattice forms and dimensions and A band spacings in relaxed, tetanized, stretched, and stretched-and-tetanized rat soleus muscle. We have tested the independent contributions of passive load, active tension, and sarcomere length to Z band state. As the A band spacing decreased with increasing load and increasing sarcomere length in the untetanized muscles, the Z lattice remained in the ss form and the Z spacing changed only slightly. Computer-enhanced images from digitized electron micrographs showed that the ss Z lattice resisted deformation regardless of load or method of stretching. In contrast, when the muscle was tetanized at sarcomere lengths of up to 2.7 microns, the Z lattice assumed the bw form and the Z spacing was increased by 20%. Regardless of lattice form, Z spacing did not vary significantly with sarcomere length. Images from freeze-substituted preparations showed both lattice forms comparable to those in images from glutaraldehyde-fixed muscles. Thus, Z band state appears to be a function of the presence (or absence) of active tension. Our previous three-dimensional model is compatible with these observations and with the sub-structures revealed by computer-enhanced images of both lattice forms.

Animals↗