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Biomedical subjects

M L Moss

Publications and source records attributed to M L Moss.

At least 19 recordsLinked to original sources
PubMed1992

Identification of actin and HSP 70 as cyclosporin A binding proteins by photoaffinity labeling and fluorescence displacement assays.

A novel family of cyclosporin A (CsA) binding proteins was identified by using the biologically active, radioiodinated photoaffinity probe [D-Lys-N epsilon-(4-azido-3-[125I]iodophenyl)propionyl)]8-CsA. In addition to cyclophilin, proteins with molecular masses of 43 kDa and approximately 50-55 kDa were labeled in Jurkat extracts and bovine calf thymus. Sequence analysis of the 43-kDa protein purified from calf thymus and subsequent Western analysis of CsA affinity-purified material from Jurkat extracts identified the 43-kDa component as actin. [D-Lys-N epsilon-(5-dimethylamino-1-naphthalenesulfonyl)]8-CsA, a fluorescent analogue of CsA, was prepared and used to measure the binding constants of cyclosporin derivatives to actin by means of a new fluorescence displacement assay. [D-Lys-N epsilon-(5-dimethylamino-1-naphthalenesulfonyl)]8-CsA and [N delta-t-butoxycarbonyl diaminobutyryl)]8-CsA bind to bovine actin at physiologically relevant concentrations, with dissociation constants of 60 +/- 33 and 570 +/- 380 nM, respectively. Because the ATPase fragment of heat shock cognate 70 (HSC 70) is structurally related to actin, the yeast homologue SSA1 was tested and found to be radiolabeled by the cyclosporin A photoaffinity reagent. The binding constant for [D-Lys-N epsilon-(5-dimethylamino-1-naphthalenesulfonyl)]8-CsA to SSA1 was determined and is 53 +/- 48 nM. These results indicate that actin and the 70-kDa heat shock protein family contain a structurally related domain for binding of cyclosporin A-related peptides.

Actins↗
PubMed1992

Fluorescence displacement method for the determination of receptor-ligand binding constants.

The equilibrium constant for the binding of a spectroscopically invisible ligand to its protein receptor can be determined in a competition experiment, by using a structural analog that contains a reporter group (fluorophor). A novel mathematical treatment of the multiple equilibria allows the analysis to be performed under tight-binding conditions. The equilibrium equation for mixtures of two mutually competitive tight-binding ligands can be expressed in a recursive form, a form in which the dependent variable appears on both sides and the solution is found iteratively. The algorithm is also applicable to the special case of weak binding, where the concentration of the bound ligand can be neglected in the mass balance. The fluorescence displacement method is demonstrated on the determination cyclophilin binding to cyclosporin A (CsA), in competition with its fluorescent derivative, [D-Lys(Dns)]8-CsA.

Amino Acid Isomerases↗
PubMed1992

When does a random flap die?

A random flap can be constructed, its circulation determined, and the ischemic portion identified. Left untreated for a period, the critical ischemia time, the ischemic portion will die and is clinically recognized several days later. What is not known is when this tissue, destined to die, actually dies. To ascertain this time, we compared the percent necrosis of a distal 3 x 3 cm segment of a 10 x 3 cm reverse McFarlane random flap with a known distribution of necrosis to the percent necrosis of the distal 3 x 3 cm of full-thickness skin grafts taken from a similar reverse McFarlane flap at 0, 4, 8, 12, and 16 hours after pedicle construction. Implicit in this experiment is the assumption that necrosis of the full-thickness skin grafts in excess of that of control animals represented skin no longer viable. Sometime between 8 and 12 hours, the percent necrosis of the full-thickness skin grafts surpassed that of the control, and it was concluded that this graft was dead prior to grafting. Thus it is suggested that critical ischemia time and death of the flap tissue are nearly identical, and the latter occurs at between 8 and 12 hours.

Animals↗
PubMed1992

Nuclear processing of the 3'-terminal nucleotides of pre-U1 RNA in Xenopus laevis oocytes.

U1 small nuclear RNA is synthesized as a precursor with several extra nucleotides at its 3' end. We show that in Xenopus laevis oocytes, removal of the terminal two nucleotides occurs after the RNA has transited through the cytoplasm and returned to the nucleus. The activity is controlled by an inhibitor of processing, which we call TPI, for 3'-terminal processing inhibitor. This inhibitor is sensitive to both micrococcal nuclease and trypsin treatment, indicating that it is a nucleoprotein. TPI inhibits the 3' processing of pre-U1 RNAs that have 5' ends containing m7G caps but not mature m2,2,7G caps; this finding suggests that TPI interacts directly or indirectly with the 5' end of pre-U1 RNA. The inhibition of processing by TPI, almost complete at 19 degrees C, is reversibly inactivated at slightly higher temperatures. TPI activity is solely in the soluble fraction of oocyte nuclear extracts, in contrast to the 3'-terminal processing activity, which is present in both the particulate and soluble fractions. We propose that the differential processing of the 3'-terminal nucleotides of pre-U1 RNA after its return from the cytoplasm, but not before its exit from the nucleus, may be due to the association of TPI with the m7G cap on the newly synthesized pre-U1 RNA.

Animals↗
PubMed1991

Epithelioid hemangioendothelioma of the lung (intravascular bronchioloalveolar tumor) in a young girl.

This report describes a 7 year old girl who presented with scoliosis and a small right hemithorax. Findings on spine roentgenographs were pleural thickening on the right and parenchymal densities. Computed tomography of the chest and abdomen revealed multiple pulmonary nodules and two hypervascular lesions within the liver. The diagnosis established by lung biopsy was intravascular bronchioloalveolar tumor (epithelioid hemangioendothelioma of the lung). This is the youngest reported case of this rare tumor.

Biopsy↗
PubMed1991

Candidates for the mechanosensory system in bone.

Some potential mechanisms by which bone cells sense mechanical loads are described and hypotheses concerning the functioning of these mechanisms are explored. It is well known that bone tissue adapts its structure to its mechanical load environment. Recent research has illuminated the biological response of bone to mechanical loading at the cellular level, but the precise mechanosensory system that signals bone cells to deposit or resorb tissue has not been identified. The purpose of this paper is to describe the current status of this research and to suggest some possible mechanosensory systems by which bone cells might sense environmental loads.

Adaptation, Physiological↗
PubMed1990

Activation of lysine 2,3-aminomutase by S-adenosylmethionine.

Lysine 2,3-aminomutase, which catalyzes the interconversion of L-lysine and L-beta-lysine, is S-adenosyl-methionine-dependent, and the adenosyl-C-5' methylene group of this coenzyme mediates the transfer of hydrogen from C-3 of lysine to C-2 of beta-lysine. We here report experiments that address the mechanism by which S-adenosylmethionine activates lysine 2,3-aminomutase. We also describe an updated and improved purification procedure that produces enzyme with a specific activity substantially higher than that previously reported. Activation of the enzyme by less than 1 mol of S-adenosyl[1-14C]methionine/mol of subunits in the presence of lysine leads to the production of [14C] methionine in a kinetically biphasic process. After 1.8 min at 30 degrees C, 10% of the 14C is reisolated as [14C] methionine, and the cleavage increases to 19% after 10 min and to 51% after 40 min. Similar experiments with S-[8-14C]adenosylmethionine produce 5'-deoxy[14C]adenosine in amounts similar to the formation of methionine. The major radioactive products isolated in each case are [14C]methionine or 5'-deoxy[14C]adenosine, respectively, and unchanged 14C-labeled S-adenosylmethionine. These experiments support the hypothesis that activation of lysine 2,3-aminomutase involves a transfer of the 5'-deoxyadenosyl moiety from S-adenosylmethionine to another species associated with the enzyme, presumably another cofactor, to form an adenosyl cofactor that functions as the proximal, hydrogen abstracting species in the mechanism.

Amino Acid Isomerases↗
PubMed1989

Lysine 2,3-aminomutase. Support for a mechanism of hydrogen transfer involving S-adenosylmethionine.

The conversion of L-lysine to L-beta-lysine is catalyzed by lysine 2,3-aminomutase. The reaction involves the interchange of the 2-amino group of lysine with a hydrogen at carbon 3. As such the reaction is formally analogous to adenosylcobalamin-dependent rearrangements. However, the enzyme does not contain and is not activated by this coenzyme. Instead it contains iron and pyridoxal phosphate and is activated by S-adenosylmethionine. Earlier experiments implicated adenosyl-C-5' of S-adenosylmethionine in the hydrogen transfer mechanism, apparently in a role similar or analogous to that of adenosyl moiety of adenosylcobalamin in the B12-dependent rearrangements. The question of whether both hydrogens or only one hydrogen at adenosyl-C-5' participate in the hydrogen-transfer process has been addressed by carrying out the lysine 2,3-aminomutase reaction with S-[5'-3H] adenosylmethionine in the presence of 10 times its molar concentration of enzyme. Under these conditions all of the tritium appeared in lysine and beta-lysine, showing that C-5'-hydrogens participate. To determine whether hydrogen transfer is compulsorily intermolecular and intramolecular, various molar ratios of [3,3-2H2]lysine and unlabeled lysine were submitted to the action of lysine 2,3-aminomutase under conditions in which 10-15% conversion to beta-lysine occurred. Mass spectral analysis of the beta-lysine for monodeutero and dideutero species showed conclusively that hydrogen transfer is both intramolecular and intermolecular. The results quantitatively support our postulate that activation of the enzyme involves a transformation of S-adenosylmethionine into a form that promotes the generation of an adenosyl-5' free radical, which abstracts hydrogen from lysine to form 5'-deoxyadenosine as an intermediate.

Amino Acid Isomerases↗
PubMed1989

Skull development in the muscular dystrophic mouse.

Roentgencephalometric tracings of skulls of 7-week-old normal and muscular dystrophic mice were compared. A marked size reduction of the dystrophic skulls relative to the normal ones was observed. However, the visceral parts of the dystrophic skull were more reduced in size than the neural parts. Marked differences in shape were also noticed. Differences in angular values were primarily found between skull parts, whereas angles between adjacent bones were remarkably similar in the two groups. Only a few exceptions of this condition were observed, as angles between adjacent bones in the posterior part of the cranial vault of the dystrophic animals differed from those of the normal animals. The observed differences between normal and dystrophic mice skulls may chiefly be explained as the results of differences in the action of diseased and normal muscles on bone.

Animals↗
PubMed1989

Fluorometric analysis of an attempt to reclaim ischemic flaps in rats with Fluosol.

An experiment was designed to answer two questions as they apply to random skin-flap survival: Is there a therapy that can improve random skin-flap survival when given postoperatively? And if so, when does one start such a therapy? Fluosol-DA 20% (Fluosol) has increased random skin-flap survival when given preoperatively in our laboratory. An experiment was devised to see if it could rescue failing flaps. One-hundred Sprague-Dawley rats were divided into a control (N = 25) and five experimental groups (N = 15). All had 10 X 13 cm reverse McFarlane random flaps raised and reinset. The experimental groups underwent hemodilution with either Ringer's lactate or Fluosol at 4, 8, and 12 hours after flap elevation. All were kept in 50% oxygen for 72 hours postoperatively. The flaps and their corresponding necrotic areas were measured on day 7. As to when to institute a therapy, we simultaneously evaluated the use of a microfluorometer as a monitor of flap survival. Analysis of flap survival showed little difference between control and experimental Ringer's lactate or Fluosol groups. Analysis of the microfluorometric data led to the following points. First, as a monitor of flap viability, it is limited by a lack of specificity and sensitivity. Second, comparison of the data from portions of the flap destined to live with those destined to die suggests that it may not be failure of circulatory inflow that leads to flap death.

Animals↗
PubMed1988

Finite element method comparison of murine mandibular form differences.

The differences of adult female murine mandibular form between two inbred strains of mice (BALB/cBy and C57BL/6By) were analyzed by use of the finite element method (FEM). In the present instance, this method overcomes the conceptual and methodologic constraints of both craniometry and roentgenographic cephalometry by providing two-dimensional, reference-frame-invariant, independent comparisons of the size, shape, and location of the point continua enclosed within the boundaries for each local, finite element into which the mandible was discretized. At the present scale of observation, it is suggested that the FEM kinematic descriptions of form comparisons can be correlated with dynamic interpretations offered by the functional matrix hypothesis, i.e., that certain discrete, causal relationships exist between the continuum form of single elements and the form, or function, of anatomic structures associated with these elements. It is concluded that, in addition to its previously demonstrated use in the analysis of growth, the finite element method seems well suited for comparisons of form.

Animals↗
PubMed1987

Studies on orthocephalization: growth behavior of the rat skull in the period 13-49 days as described by the finite element method.

Rat cranial skeletal growth was studied, using a cross-sectional data set, for the period 13-49 days by the application of the concepts of continuum mechanics and the numerical techniques of the finite element method (FEM). In contrast to the methods of conventional craniometry (CM) and roentgenographic cephalometry (RCM) the FEM permits fine scale, reference frame invariant descriptions and analysis of growth behavior. This advantage was demonstrated by a numerical example of the use of FEM. The skull was discretized into a number of two-dimensional, triangular elements, whose enclosed areas corresponded closely to both specific skeletal structures and to related functional matrices. Since it was assumed presently that the growth behavior of all of the points enclosed within a given element was similar, the application of the functional matrix hypothesis permitted an integrated description of the growth of the skeletal structure and functional matrix related to each element. The principal locus of rotation of the facial skull, relative to the cranial base, is the inferior frontoethmoidal articulation, a motion that includes a rigid body rotation. Other active and passive skeletal and visceral growth events associated with orthocephalization were located and described. Finally it was shown that the morphogenetically important growth behavior of other portions of the rat head were not directly involved in orthocephalization.

Aging↗
PubMed1987

Fluorocarbon enhancement of skin flap survival in rats.

Fluorocarbons exhibit two unique properties: oxygen saturation in direct proportion to the percent administered and low viscosity which improves microcirculation. These properties were investigated in improving survival in random skin flaps in rats. Modified McFarlane flaps were raised in 30 Sprague-Dawley rats and divided into three equal groups. Group 1 rats served as controls, group 2 rats were hemodiluted with Ringer's lactate, and group 3 rats were hemodiluted with Fluosol-DA (20%). All groups were kept in a high (80%) oxygen environment for 48 hours. Areas of necrosis were measured using a computer system. Necrosis in control flaps averaged 14.96 percent; in flaps hemodiluted with Ringer's lactate, 10.12 percent; in flaps hemodiluted with Fluosol, 4.76 percent. These differences were statistically significant. We conclude that fluorocarbons significantly enhance flap survival in rats.

Animals↗
PubMed1987

Effects of pre-weaning undernutrition on 21 day-old male rat skull form as described by the finite element method.

Conventional Roentgenographic Cephalometric Methods (RCM) have certain conceptual and geometric constraints that ensure that their descriptions of cephalic growth and comparisons of cephalic form are reference-frame-dependent; and it is impossible to determine which, if any, RCM reference frame provides a biologically more correct description or comparison. The use of the concepts of continuum mechanics and of the numerical techniques of the Finite Element Method (FEM) overcomes these constraints and provides reference-frame-independent (invariant) growth descriptions and form comparisons. In the FEM, the structure of the head is subdivided (discretized) into a number of smaller, finite, elements. The FEM then describes the growth behavior, or compares the form, of the entire continuum of points enclosed with each element independently; in RCM only the behaviors, or comparisons, of the landmark points are described. The FEM is described and its use illustrated by a comparison of the cephalic forms of 4 groups of weanling (21 d old) rats. The 1st (control) group is derived from normally nourished dams, while the other 3 groups are derived from dams who were malnourished throughout the weaning period in 3 group specific manners. The FEM permitted reference-frame-independent comparisons of the 4 different, and group specific, head forms. These differences were reflected in the forms of each of the finite elements. The differences in element forms between the 4 groups were shown to be related primarily to nutritionally produced alterations of either neurocranial or splanchocranial viscera. These differences were expressed in the alterations of the forms of the continua enclosed within the boundaries of the several finite elements, and secondarily in the form of the cranial skeleton.

Animals↗
PubMed1987

Morphological analysis and computer-aided, three dimensional reconstruction of chondrocytic columns in rabbit growth plates.

Proximal tibial growth plates of New Zealand white rabbits were serially sectioned in parasagittal and horizontal planes for three dimensional, light microscopic analysis of the chondrocytic columns. A total of 431 columns was analysed. Of these, 258 columns extended through the full height of the growth plate. The remaining columns were considerably shorter, being located either predominantly in the epiphyseal half of the growth plate (100) or in the metaphyseal half of the growth plate (73). The epiphyseal and metaphyseal columns were found in clusters in the plate. Some columns in all three groups had interruptions along their length, while others had duplications. Computer-aided, three dimensional graphic reconstructions were prepared of a selected group of columns. The reconstructions illustrated the variability in the morphology and the dimensions of the neighbouring chondrocytic columns. The observations suggest that chondrocytic columns in rabbit growth plates are replaced regularly and that the small cell zone may play an important role as the cellular source for column renewal.

Animals↗
PubMed1987

Studies on orthocephalization. 10. Behaviour of the visceral part of the rat head during the first 14 days after gestation.

The present paper considers the significance of interosseous flexions of the palatal complex in the process of orthocephalization of the rat skull between birth and 7 d p.n. The study is based on a sample of 90 rats divided into 4 age groups, i.e. 0, 4, 7, and 14 d. These rats have been X-rayed, and their photographs subsequently analysed. During the studied period, the constituents of the bony palate, i.e. the horizontal part of the palatine bone, the palatal process of maxilla and the palatal part of premaxilla, increase markedly in length, but with individual differences in growth rate. There is, in the period, a marked decrease in angulation between the cranial base and the palatal plane. This means that the rat skull becomes more orthocranial. There is also a straightening (orthopalatalization) of the palate, as the angle between maxilla and premaxilla becomes more obtuse, and a marked decrease in angulation between the palatine bone and the cranial base. The patterns of angular changes suggest that the process of orthocephalization in the period between birth and 14 d p.n. primarily is a result of an upwards rotation of the palatine bone relative to the cranial base, while interosseous deflections in the palate only play a minor role.

Animals↗