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R D Goldman

Publications and source records attributed to R D Goldman.

At least 37 records · Page 2Linked to original sources
PubMed2000

The 300-kDa intermediate filament-associated protein (IFAP300) is a hamster plectin ortholog.

Plectin is a high-molecular-weight cytoskeleton-associated protein that was initially identified in intermediate filament (IF)-enriched fractions of rat C6 glioma cells. At the cellular level, plectin has been found to associate with IF networks and IF-associated structures that are involved in cell-cell and cell-substrate adhesions. IFAP300 is an IF-associated protein that was initially identified in hamster cells by a monoclonal antibody directed against a high molecular weight protein present in IF-enriched cytoskeletal preparations. Plectin and IFAP300 display similar distribution patterns within cells as determined by immunofluorescence. Based upon this and the finding that their biochemical properties are similar, it has been suggested that they may actually be orthologous proteins. In this paper we demonstrate that this is the case. Cloning and sequencing of most of the hamster plectin cDNA demonstrates that plectin is found in hamster cells and that its sequence is highly conserved between species. Using immunological cross-reactivity, epitope mapping, and immunoelectron microscopy, we show that IFAP300 is actually the hamster ortholog of plectin.

Amino Acid Sequence↗
PubMed2000

Disruption of nuclear lamin organization blocks the elongation phase of DNA replication.

The role of nuclear lamins in DNA replication is unclear. To address this, nuclei were assembled in Xenopus extracts containing AraC, a reversible inhibitor that blocks near the onset of the elongation phase of replication. Dominant-negative lamin mutants lacking their NH(2)-terminal domains were added to assembled nuclei to disrupt lamin organization. This prevented the resumption of DNA replication after the release of the AraC block. This inhibition of replication was not due to gross disruption of nuclear envelope structure and function. The organization of initiation factors was not altered by lamin disruption, and nuclei resumed replication when transferred to extracts treated with CIP, an inhibitor of the cyclin-dependent kinase (cdk) 2-dependent step of initiation. This suggests that alteration of lamin organization does not affect the initiation phase of DNA replication. Instead, we find that disruption of lamin organization inhibited chain elongation in a dose-dependent fashion. Furthermore, the established organization of two elongation factors, proliferating cell nuclear antigen, and replication factor complex, was disrupted by DeltaNLA. These findings demonstrate that lamin organization must be maintained in nuclei for the elongation phase of DNA replication to proceed.

Animals↗
PubMed2000

Rapid displacement of vimentin intermediate filaments in living endothelial cells exposed to flow.

Hemodynamic shear stress at the endothelial cell surface induces acute and chronic intracellular responses that regulate vessel wall biology. The cytoskeleton is implicated by acting both as a direct connector to local surface deformation and as a distribution network for mechanical forces throughout the cell; however, direct observation and measurement of its position during flow have only recently become possible. In this study, we directly demonstrate rapid deformation of the intermediate filament (IF) network in living endothelial cells subjected to changes in hemodynamic shear stress. Time-lapse optical sectioning and deconvolution microscopy were performed within the first 3 minutes after the introduction of flow (shear stress, 12 dyn/cm(2)). Spatial and temporal dynamics of green fluorescent protein-vimentin IFs in confluent endothelial cells were analyzed. The imposition of shear stress significantly increased the variability of IF movement throughout the cell in the x-, y-, and z-directions compared with the constitutive dynamics noted in the absence of flow. Acute polymerization and depolymerization of the IF network were absent. The magnitude and direction of flow-induced IF displacement were heterogeneous at the subcellular level. These qualitative and quantitative data demonstrate that shear stress acting at the luminal surface of the endothelium results in rapid deformation of a stable IF network.

Animals↗
PubMed2000

Review: the dynamics of the nuclear lamins during the cell cycle-- relationship between structure and function.

The nuclear lamins are members of the intermediate filament (IF) family of proteins. The lamins have an essential role in maintaining nuclear integrity, as do the other IF family members in the cytoplasm. Also like cytoplasmic IFs, the organization of lamins is dynamic. The lamins are found not only at the nuclear periphery but also in the interior of the nucleus, as distinct nucleoplasmic foci and possibly as a network throughout the nucleus. Nuclear processes such as DNA replication may be organized around these structures. In this review, we discuss changes in the structure and organization of the nuclear lamins during the cell cycle and during cell differentiation. These changes are correlated with changes in nuclear structure and function. For example, the interactions of lamins with chromatin and nuclear envelope components occur very early during nuclear assembly following mitosis. During S-phase, the lamins colocalize with markers of DNA replication, and proper lamin organization must be maintained for replication to proceed. When cells differentiate, the expression pattern of lamin isotypes changes. In addition, changes in lamin organization and expression patterns accompany the nuclear alterations observed in transformed cells. These lamin structures may modulate nuclear function in each of these processes.

Animals↗
PubMed2000

Risk for one or two very low birth weight twins: a population study.

OBJECTIVE: To calculate the frequencies of very low birth weight (VLBW) neonates among twins in a large population database. METHODS: The database comprised 12,567 live-born twin pairs delivered from 1993 to 1998 in Israel. Low birth weight (LBW) and VLBW were defined as less than 2500 and 1500 g, respectively. We counted the number of pairs with VLBW neonates in three combinations: VLBW-VLBW, VLBW-LBW, and VLBW-over 2500 g. We compared the subsets of nulliparas and multiparas and the frequency of like- versus unlike-sex twins. RESULTS: The frequency of at least one VLBW twin was significantly higher among nulliparas than multiparas (odds ratio [OR] 2.3; 95% confidence interval [CI] 2.1, 2.6; P <.001). For pairs with VLBW-VLBW and VLBW-LBW combinations, a significantly higher frequency was found among nulliparas than multiparas (OR 2.0; 95% CI 1.7, 2.8; P <.001 and OR 2.6; 95% CI 2.2, 3.1; P <.001, respectively). The risk seemed to be accentuated in like-sex twins. Overall, the risk of having at least one VLBW infant was 1:5 among nulliparas and 1:12 among multiparas. The risk of having two VLBW twins among nulliparas (1:11) was double that of multiparas (1:22). CONCLUSION: Nulliparas are at significantly increased risk of delivering one or two VLBW twins.

Cross-Sectional Studies↗
PubMed2000

Adaptive growth restriction as a pattern of birth weight discordance in twin gestations.

OBJECTIVE: To challenge the hypothesis that discordant growth is a normal variation by relating birth weight discordance to total twin birth weight. METHODS Among 12,565 Israeli live-born twin pairs (1993-98), we compared total twin birth weight decile, the frequencies of three levels of discordance in the general population, over 25% discordance between like- versus unlike-sex pairs, and over 25% discordance between pairs delivered by primiparas versus multiparas. RESULTS: We found a marked change in the best-fit correlation function with increased discordance: level 15-24.9% was inversely linear whereas levels 25-34.9% and over 34.9% were inversely logarithmic (R(2) =.47,.88, and.9, respectively). The best-fit correlation of frequencies of more than 25% discordance was inversely logarithmic and similar in like- and unlike-sexed twins across deciles functions (P =.7, odds ratio [OR] 1.0, 95% confidence interval [CI] 0.9, 1.2). The overall frequencies of discordance were also similar (9.3% versus 10.2%, P =.11, OR 0.9, 95% CI 0.8, 1.0). The frequencies of primiparas decreased linearly (R(2) =.98) and the frequencies of more than 25% discordant pairs in multiparas and primiparas across the deciles had similar inversely logarithmic patterns (P =.55, OR 1.0, 95% CI 0.9, 1.2). Discordance over 25% was significantly more frequent among primiparas (P <.001, OR 1.45, 95% CI 1.3, 1.6). CONCLUSION: The observed patterns of birth weight discordance did not substantiate normal variation but an adaptive growth restriction that might explain why the likelihood of discordant growth decreases as total twin birth weight increases.

Adaptation, Physiological↗
PubMed2000

Delivery of breech first twins: a multicenter retrospective study.

OBJECTIVE: To assess the risk of vaginal birth of breech first twins by Apgar scores and mortality. METHODS: We did a retrospective case-control analysis of data from 13 centers that allow vaginal birth for breech first twins. We used depressed 5-minute Apgar scores and neonatal mortality as main outcome measures between vaginal (n = 239) and cesarean (n = 374) deliveries of pairs with breech first twins, stratified by parity, birth weights of first twins, and types of cesarean. The 95% power of our sample size (alpha = .05) was sensitive enough to detect differences of 5% of the overall sample and 25-30% of subgroups. RESULTS: Vaginal birth was attempted in 61% of 613 pairs. There were significantly more depressed Apgar scores (P = .008, odds ratio [OR] 2.4, 95% confidence interval [CI] 1.2, 4.7) and neonatal deaths (P < .001, OR 9.5, 95% CI 4.0, 23.4) among vaginal births of pairs in whom first twins weighed less than 1500 g but not among the higher-birth-weight cohort (for depressed Apgar scores: P = .76, OR 1.1, 95% CI 0.6, 2.1). Multiparity and elective cesarean seemed to have little influence on outcome measures. Neonatal mortality was associated with extremely preterm twins. CONCLUSION: There was no evidence that vaginal birth is unsafe, in terms of depressed Apgar scores and neonatal mortality, for breech first twins that weighed at least 1500 g.

Apgar Score↗
PubMed2000

Fast transport of neurofilament protein along microtubules in squid axoplasm.

Using squid axoplasm as a model system, we have visualized the fast transport of non-filamentous neurofilament protein particles along axonal microtubules. This transport occurs at speeds of 0.5-1.0 microm/second and the majority of neurofilament particles stain with kinesin antibody. These observations demonstrate, for the first time, that fast (0.5-1.0 microm/second) transport of neurofilament proteins occurs along microtubules. In addition, our studies suggest that neurofilament protein can be transported as non-membrane bound, nonfilamentous subunits along axons, and that the transport is kinesin-dependent. Microtubule-based fast transport might therefore provide a mechanism for the distribution and turnover of neurofilament, and perhaps other cytoskeletal proteins, throughout neurons.

Amino Acid Sequence↗
PubMed1999

Integrating the actin and vimentin cytoskeletons. adhesion-dependent formation of fimbrin-vimentin complexes in macrophages.

Cells adhere to the substratum through specialized structures that are linked to the actin cytoskeleton. Recent studies report that adhesion also involves the intermediate filament (IF) and microtubule cytoskeletons, although their mechanisms of interaction are unknown. Here we report evidence for a novel adhesion-dependent interaction between components of the actin and IF cytoskeletons. In biochemical fractionation experiments, fimbrin and vimentin coprecipitate from detergent extracts of macrophages using vimentin- or fimbrin-specific antisera. Fluorescence microscopy confirms the biochemical association. Both proteins colocalized to podosomes in the earliest stages of cell adhesion and spreading. The complex is also found in filopodia and retraction fibers. After detergent extraction, fimbrin and vimentin staining of podosomes, filopodia, and retraction fibers are lost, confirming that the complex is localized to these structures. A 1:4 stoichiometry of fimbrin binding to vimentin and a low percentage (1%) of the extracted vimentin suggest that fimbrin interacts with a vimentin subunit. A fimbrin-binding site was identified in the NH(2)-terminal domain of vimentin and the vimentin binding site at residues 143-188 in the CH1 domain of fimbrin. Based on these observations, we propose that a fimbrin-vimentin complex may be involved in directing the assembly of the vimentin cytoskeleton at cell adhesion sites.

Actins↗
PubMed1999

A high molecular weight intermediate filament-associated protein in BHK-21 cells is nestin, a type VI intermediate filament protein. Limited co-assembly in vitro to form heteropolymers with type III vimentin and type IV alpha-internexin.

BHK-21 fibroblasts contain type III vimentin/desmin intermediate filament (IF) proteins that typically co-isolate and co-cycle in in vitro experiments with certain high molecular weight proteins. Here, we report purification of one of these and demonstrate that it is in fact the type VI IF protein nestin. Nestin is expressed in several fibroblastic but not epithelioid cell lines. We show that nestin forms homodimers and homotetramers but does not form IF by itself in vitro. In mixtures, nestin preferentially co-assembles with purified vimentin or the type IV IF protein alpha-internexin to form heterodimer coiled-coil molecules. These molecules may co-assemble into 10 nm IF provided that the total amount of nestin does not exceed about 25%. However, nestin does not dimerize with types I/II keratin IF chains. The bulk of the nestin protein consists of a long carboxyl-terminal tail composed of various highly charged peptide repeats. By analogy with the larger neurofilament chains, we postulate that these sequences serve as cross-bridgers or spacers between IF and/or other cytoskeletal constituents. In this way, we propose that direct incorporation of modest amounts of nestin into the backbone of cytoplasmic types III and IV IFs affords a simple yet flexible method for the regulation of their dynamic supramolecular organization and function in cells.

Animals↗
PubMed1999

The relation between inter-twin birth weight discordance and total twin birth weight.

OBJECTIVE: We tested the hypothesis that the frequency of growth discordance among twins is not related to the uterine capacity for carrying twins. METHOD: We counted and compared the frequencies of birth weight discordance of more than 25% in an unlike-sexed twin cohort (n = 1244) and in a population-based twin cohort (n = 7570) across the deciles of the total twin birth weight (twin A + twin B) distribution. The birth order of the heavier twin was noted. RESULTS: Similar frequencies of discordant pairs were found in both cohorts (11% and 12%, respectively; Mantel-Haenszel chi2 test: P = .131, odds ratio (OR) 0.9, 99% confidence interval (CI) 0.67, 1.11; Woolf test for heterogeneity: two-tailed P = .472). In the discordant pairs, twin A was considerably more often the heavier twin in all birth weight deciles (unlike-sexed cohort: P < 10(-8), OR 5.9, 99% CI 3.0, 11.7; population-based cohort: P < 10(-8), OR 3.1, 99% CI 2.3, 4.0), and in both cohorts (inter-cohort difference: P = .109, OR 1.4, 99% CI 0.83, 2.32). Both cohorts showed a similar nonlinear trend: given that X = decile order, discordance decreased as a function of 22.0 - 6.54 ln[X] for the unlike-sexed twins cohort and 23.0 - 8.18 ln[X] for the population-based cohort, with r values of 0.967. CONCLUSION: The more favorable the uterine milieu for carrying twins, the smaller the likelihood of discordant twin growth. Birth order of the heavier twin appears to be an integral part of the discordance phenomenon. The similarity of the cohorts suggests that these conclusions are valid for both like and unlike-sexed twins.

Birth Weight↗
PubMed1999

Parental estimates of their child's weight: accurate for resuscitation drug doses.

OBJECTIVE: The aim of this study was to assess parental reliability in estimating child body weight in emergency situations, when weighing the child is often impossible. METHODS: 312 parents were asked to complete an anonymous questionnaire that included estimating their child's weight. 233 questionnaires were enrolled in the study and were assessed statistically using Students t test, and chi2 and ANOVA tests. RESULTS: 51.5% of parents estimated their child's body weight within +/-5% of the true weight, 73.4% within +/-10%, and 87.5% within +/-20%. A significant difference was found between paternal and maternal estimations, with 56% of mothers and only 40.3% of fathers estimating within a 5% range of accuracy (P < 0.05). CONCLUSIONS: Parents, especially mothers, can estimate their child's body weight within clinically acceptable limits. These estimations can reliably be used to calculate drug doses in prehospital and emergency department situations, when children's weight is not known and cannot be measured.

Anthropometry↗
PubMed1998

Motile properties of vimentin intermediate filament networks in living cells.

The motile properties of intermediate filament (IF) networks have been studied in living cells expressing vimentin tagged with green fluorescent protein (GFP-vimentin). In interphase and mitotic cells, GFP-vimentin is incorporated into the endogenous IF network, and accurately reports the behavior of IF. Time-lapse observations of interphase arrays of vimentin fibrils demonstrate that they are constantly changing their configurations in the absence of alterations in cell shape. Intersecting points of vimentin fibrils, or foci, frequently move towards or away from each other, indicating that the fibrils can lengthen or shorten. Fluorescence recovery after photobleaching shows that bleach zones across fibrils rapidly recover their fluorescence. During this recovery, bleached zones frequently move, indicating translocation of fibrils. Intriguingly, neighboring fibrils within a cell can exhibit different rates and directions of movement, and they often appear to extend or elongate into the peripheral regions of the cytoplasm. In these same regions, short filamentous structures are also seen actively translocating. All of these motile properties require energy, and the majority appear to be mediated by interactions of IF with microtubules and microfilaments.

Animals↗
PubMed1998

Rapid movements of vimentin on microtubule tracks: kinesin-dependent assembly of intermediate filament networks.

The assembly and maintenance of an extended intermediate filament (IF) network in fibroblasts requires microtubule (MT) integrity. Using a green fluorescent protein-vimentin construct, and spreading BHK-21 cells as a model system to study IF-MT interactions, we have discovered a novel mechanism involved in the assembly of the vimentin IF cytoskeleton. This entails the rapid, discontinuous, and MT-dependent movement of IF precursors towards the peripheral regions of the cytoplasm where they appear to assemble into short fibrils. These precursors, or vimentin dots, move at speeds averaging 0.55 +/- 0.24 micrometer/s. The vimentin dots colocalize with MT and their motility is inhibited after treatment with nocodazole. Our studies further implicate a conventional kinesin in the movement of the vimentin dots. The dots colocalize with conventional kinesin as shown by indirect immunofluorescence, and IF preparations from spreading cells are enriched in kinesin. Furthermore, microinjection of kinesin antibodies into spreading cells prevents the assembly of an extended IF network. These studies provide insights into the interactions between the IF and MT systems. They also suggest a role for conventional kinesin in the distribution of non-membranous protein cargo, and the local regulation of IF assembly.

Animals↗
PubMed1998

The intermediate filament protein, vimentin, in the lens is a target for cross-linking by transglutaminase.

Mere addition of Ca2+ to a lens cortical homogenate (bovine) generates a series of products composed of a variety of high molecular weight vimentin species. The Ca2+-induced cross-linking of this cytoskeletal element seems to be mediated by the intrinsic transglutaminase of lens, because the reaction could be blocked at the monomeric state of vimentin by the inclusion of small synthetic substrates of the enzyme dansylcadaverine or dansyl-epsilon-aminocaproyl-Gln-Gln-Ile-Val. These compounds are known to compete against the Gln or Lys functionalities of proteins that would participate in forming the Nepsilon(gamma-glutamyl)lysine protein-to-protein cross-links. The cytosolic transglutaminase-catalyzed reactions could be reproduced with purified bovine lens vimentin and also with recombinant human vimentin preparations. Employing the latter system, we have titrated the transglutaminase-reactive sites of vimentin and, by sequencing the dansyl-tracer-labeled segments of the protein, we have shown that residues Gln453 and Gln460 served as acceptor functionalities and Lys97, Lys104, Lys294, and Lys439 as electron donor functionalities in vimentin. The transglutaminase-dependent reaction of this intermediate filament protein might influence the shape and plasticity of the fiber cells, and the enzyme-catalyzed cross-linking of vimentin, in conjunction with other lens constituents, may contribute to the process of cataract formation.

Amino Acid Sequence↗