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Effect of structural change on acute toxicity and antiinflammatory activity in a series of imidazothiazoles and thiazolobenzimidazoles.

The effect of structural change on the biological activity of a series of imidazothiazoles and thiazolobenzimidazoles is described. It was found that compounds with polar substituents at the 2 or 3 position of the ring system are less acutely toxic while maintaining antiinflammatory activity. Other structural changes, such as the incorporation of a gem-dimethyl substituent in the 6 position, increase acute toxicity and eliminate antiinflammatory activity. The compound with the best activity/toxicity ratio contains an alkyl sulfonyl substituent on the thiazole ring. The thiazolobenzimidazole analogues are more potent than the imidazole analogues.

Animals

Relationship between initial cardiovascular structural changes and daytime and nighttime blood pressure monitoring.

It has been shown that in hypertensive patients the degree of target organ damage correlates more closely with average blood pressure as recorded by ambulatory monitoring (ABPM) throughout 24 h than with clinic blood pressure. We examined a group of 91 clinically healthy subjects, 23 normotensives and 68 hypertensives according to clinic blood pressure. Cardiac anatomy was investigated by echocardiography. As an index of arterial structural changes forearm minimal vascular resistance was calculated from mean arterial pressure and maximal postischemic blood flow, as assessed by venous occlusion plethysmography. The results were correlated to clinic blood pressure or ABPM values (measured by noninvasive ABPM ICR 5200, Spacelabs, Bellevue, CA). Left ventricular mass was correlated more closely with the average blood pressure recorded during 24 h, or during daytime or nighttime periods, than with clinic blood pressure. Minimal vascular resistance was also significantly correlated to ABPM values, but the correlation was similar to that observed with clinic blood pressure. Minimal vascular resistance was significantly correlated to blood pressure variability, as evaluated by the standard deviation of the mean. Minimal vascular resistance and left ventricular mass were higher in a subgroup of patients in whom blood pressure was not significantly reduced during the night. The results of this study confirm that elevated average ABPM values are associated to higher left ventricular mass; in addition, they suggest that increased blood pressure variability may be associated with vascular structural changes, as evaluated by minimal vascular resistance. It remains to be clarified whether cardiac hypertrophy and/or vascular structural changes are a cause or consequence of increased blood pressure values and variability.

Adult

Humoral trophic influence on cardiovascular structural changes in hypertension.

Since the early development of structural cardiovascular change in spontaneously hypertensive rats (SHR) and stroke-prone SHR (SHRSP) indicated the involvement of non-pressure-dependent factors in this process in hypertension, smooth muscle cells (SMC) from the aorta of SHR, SHRSP, and normotensive Wistar-Kyoto rats (WKY) were investigated under tissue culture conditions free from blood pressure and humoral factors in vivo. By the observation of such factors as growth rate and DNA or protein synthesis vascular SMC from these rats with genetic hypertension were proved to have intrinsically greater growth activity independently of blood pressure. Although serum from SHR and SHRSP had no specific stimulative effect on SMC growth, circulating epinephrine may accelerate cardiovascular structural changes because isoproterenol added to the culture media enhanced ornithine decarboxylase (ODC) activity. Moreover, SMC from SHR and SHRSP showed greater thymidine incorporation than those from WKY even in response to lower extracellular Na+ concentration. Local nutritional conditions of SMC, which were proved to have a great effect on the morphology and structure of cultured SMC, may be a basic determinant of the development of hypertension-induced structural vascular changes or lesions.

Animals

A fluorescence probe of energy-dependent structure changes in fragmented membranes.

The reaction of the fluorochrome, 8-anilino-1-naphthalene-sulfonic acid (ANS), with fragmented membranes from beef heart mitochondria has been studied. ANS fluorescence is found to be enhanced 25-fold on binding to the membrane fragments in the absence of energy conservation, and this enhancement is increased to 35-fold in the membrane energized by substrate plux oxygen. The fluorescence of bound ANS depends upon the energy state of the membrane fragments, as indicated by the effects of ATP, substrates of the respiratory chain, oligomycin, and uncouplers. It is concluded that the changes of ANS fluorescence indicate structural changes of the mitochondrial membrane associated with energy conservation. The time course of energization is readily followed by ANS, and has a half-time of two seconds at 26 degrees .

Adenosine Triphosphate

D-ribulose-1,5-bisphosphate carboxylase/oxygenase: function-dependent structural changes.

The key carboxylating enzyme of the reductive pentose phosphate cycle, D-ribulose-1,5-bisphosphate carboxylase/oxygenase [RuBisCO] isolated from the chemolithoautotrophic, H2-oxidizing bacterium Alcaligenes eutrophus H16 has been analyzed by several different techniques that allow conclusions about structure and function-dependent structural changes. The techniques include a novel approach in which the enzyme was induced to form 2D-crystals suitable for electron microscopy in each of its three stable functional states: as active enzyme [Ea] (in the presence of Mg2+ and HCO3-); as inactivated enzyme [Eia] (in the absence of Mg2+ and HCO3-) and as enzyme locked in an in vitro transition state [CABP-E] (Ea fully saturated with the transition state analogue 2-carboxy-D-arabinitol-1,5-bisphosphate [CABP-E]). In conjunction with X-ray crystallography, X-ray small angle scattering and other biophysical and biochemical data, the results obtained by electron microscopy support the idea that drastic configurational changes occur. Upon transition from Ea to the CABP-E the upper and lower L4S4 halves of the molecule consisting of eight large and eight small subunits (L8S8; MW = 536,000 Da) are assumed to be laterally shifted by as much as 3.6 nm relative to one another while the location of the small subunits on top of the large subunits, and relative to them, remains the same. For the Eia a similar sliding-layer configurational change in the range of 2-2.5 nm is proposed and in addition it is suggested that other configurational/conformational changes take place. The proposed structural changes are discussed with respect to the current model for the tobacco enzyme and correlated with data obtained for various other plant and (cyano) bacterial L8S8 RuBisCOs leading to speculations about structure-function relationships.

Microscopy, Electron

Control of cell differentiation during proliferation. II. Myeloid differentiation and cell cycle arrest of HL-60 promyelocytes preceded by nuclear structural changes.

The time-dependent dynamics of nuclear structure, cell cycle transit and arrest, and cellular differentiation were studied using the human promyelocytic leukemia cell line HL-60. Myeloid differentiation of HL-60 cells was induced by 10(-6) M beta, all trans, retinoic acid (RA). During exponential growth the cells had G1, S, G2 and M durations of 9, 11, 0.5 and 0.5 h respectively. Significant growth arrest in the G1/0 phase of the cell cycle was apparent after 48 h of RA exposure or after two division cycles. Thereafter, cells arrested in G1/0 with wide dispersion in times of arrest which extended over several cell cycle generation times. The kinetics of phenotypic differentiation, detected by phorbol myristate acetate inducible superoxide production, paralleled those of G1/0 growth arrest with similar lag and dispersion. These kinetics are consistent with a model hypothesizing the existence of an S-phase differentiation control (DC) point regulating both terminal proliferation and differentiation. Before any cell differentiation or termination of cell proliferation occurred, the nuclei of RA-treated cells underwent a structural change detected by narrow-angle light scatter measured with flow cytometry. Narrow-angle light scatter was transiently reduced, reaching a nadir at 24-48 h and returning to control values at 96 h. This change was independent of cell cycle phase or total nuclear protein content. It was associated with a morphological change of the nuclear membrane from a smooth to dimpled or pitted structure. These findings focus attention on the potential significance of nuclear structural reorganization as an early event during cell differentiation.

Cell Cycle

Ca(2+)-dependence of structural changes in troponin-C in demembranated fibers of rabbit psoas muscle.

The Ca(2+)-dependence of structural changes in troponin-C (TnC) has been detected by monitoring the fluorescence from TnC labeled at Methionine-25, in the NH2-terminal domain, with danzylaziridine (TnC-DANZ) and then exchanged for endogenous TnC in glycerinated single fibers. The fluorescence-pCa relation obtained from fibers stretched to a sarcomere length greater than 4.0 microns evidenced two transitions: a small one, attributable to the binding of Ca2+ to the high affinity, Ca(2+)-Mg(2+)-binding sites of TnC; and a large one, attributable to the binding of Ca2+ to the low affinity, Ca(2+)-specific binding sites of TnC. In the fluorescence-pCa relation determined with fibers set to a sarcomere length of 2.4 microns, hence obtained in the presence of cycling cross-bridges, the large transition had the same Ca(2+)-dependence as did the development of tension. These results indicate that the NH2-terminal globular domain of TnC is modified by the binding of Ca2+ to sites located in both globular domains and that the structural changes in TnC resulting from the binding of Ca2+ to the low-affinity sites, but not to the high-affinity sites, are directly associated with the triggering of contraction.

Adenosine Triphosphate

The effects of protein synthesis inhibition on structural changes associated with learning in the chick.

The effect of the protein synthesis inhibitor anisomycin on the structural changes associated with passive avoidance learning in the chick was investigated. Chicks were trained when they were 24 h old by allowing them to peck at a shiny bead coated with either water or the aversive-tasting substance methylanthranilate (MeA). Chicks which peck the MeA-coated bead will on subsequent testing avoid pecking a similar, but water-coated bead. Behavioural testing was carried out 12 h after training and immediately afterwards the chicks were killed and their brains prepared for electron microscopy. A specific region of the forebrain, the intermediate and medial part of the hyperstriatum ventrale (IMHV) was investigated. When the IMHV of the MeA trained chicks was compared with that of water-trained controls structural changes of the synapse were detected. These changes involved a significant increase in the mean length of the postsynaptic density (LPSD) of symmetrical synapses in the left IMHV. Chicks injected with 0.8 mg of anisomycin 30 min before training with a MeA-coated bead showed aversion for the shiny bead when tested 12 h later. Electron microscopic analysis of the IMHV from these amnestic chicks showed no evidence for the change in LPSD demonstrated in the water-injected controls. These results are discussed in relation to the nature of the memory trace induced by training on a passive avoidance task.

Animals

Structural changes in the canine lung and pulmonary arteries after pneumonectomy.

The study set out to describe the structural changes in the remaining lung of beagle dogs after left-sided pneumonectomy, performed at 6-10 wk or 1 yr of age. After a 5-yr postoperative recovery, lung volume and weight had increased in the operated groups, though only the latter was significant in comparison with 5-yr-old controls. In both operated groups, total alveolar number in the right lung was normal. The total volume of alveolar ducts, mean linear intercept, and (in the young operated group) mean alveolar volume were increased, indicating a compensatory enlargement of distal air spaces. Lobar differences reinforced these conclusions. In the 1-yr operated group, medial hypertrophy was present in preacinar arteries, with a similar trend in intra-acinar arteries where peripheral extension of medial smooth muscle was also present. Despite the lack of structural change in their pulmonary arterial system, the young operated group had right ventricular hypertrophy. Previously reported evidence of alveolar proliferation shortly after pneumonectomy probably represents a speeding up of normal developmental patterns, but after prolonged recovery the final alveolar number is no greater than normal.

Animals

Immunohistochemical studies on structural changes of the hepatic lobules in chronic liver diseases.

Liver biopsy specimens were examined immunohistochemically to clarify structural changes of the hepatic lobules in chronic liver diseases. In normal liver carbohydrate antigen 19-9 was located in the biliary ductular epithelium, whereas factor VIII-related antigen was observed in the endothelium of portal veins, hepatic arteries, and central veins. This antigen was not detected in the sinusoidal endothelium. In contrasts, monoclonal antibody OKM5 was reactive with the sinusoidal endothelium but was unreactive with the endothelium of the portal blood vessels or central veins. In chronic active hepatitis and liver cirrhosis, both carbohydrate antigen 19-9 positive biliary ductular cells and factor VIII-related antigen positive endothelial cells were not only observed in the enlarged portal area but also extended into the parenchyma. They were occasionally accompanied by fibers. These findings suggest that fibrosis, ductular epithelial, and blood vascular proliferation in the portal space and their invasion into the parenchyma might gradually cause structural changes of the hepatic lobules in chronic liver disease.

Antigens

Structure-activity relationship between prostacyclin and its platelet receptor. Correlation of structure change and the platelet activity.

Correlation analysis between the structural changes of PGI2 and the corresponding changes in platelet activity was performed to look into the following structural features: (1) the spatial relationship of critical functional groups, (2) conformational flexibility of the molecule as a result of chemical modification; (3) charge distribution around C6a; (4) hydrogen-bonding capability and mispairing and (5) the steric effects which resulted from chemical modifications. Our studies led to three important conclusions: (1) The C1 carboxylate and C11, C15 hydroxyl groups of PGI2 are essential for platelet activity. Modifications that change their relative positions reduce the activity. The ring structure and the C5 and C13 double bonds are molecular designs to maintain this unique geometry. (2) The C6a oxygen, although not vital to the binding geometry, is important for the biological potency. Decreasing the electronegativity of C6a oxygen leads to decreased potency. (3) We propose that any additional hydrogen-bond donor present between C1 and C15 could cause a hydrogen-bond mispairing and therefore a decreased activity. These findings should be useful for designing new PGI2 analogues and for determining the configuration of receptor-associated PGI2 by a molecular mechanics technique.

Animals

Structural changes in the tracheae of preterm lambs induced by ventilation.

Compliant immature airways sustain significant deformation following positive pressure ventilation. To evaluate the structural changes induced by in vivo positive pressure ventilation, tracheae of preterm lambs (107-116 d gestational age) were studied histologically. Nonventilated (group I: n = 7) and ventilated (group II: n = 7) tracheal segments were excised and studied by histologic and morphometric techniques. Computerized image analysis was used to measure dimensions of tracheal wall components and of the tracheal section. The circumference, diameter, and cross-sectional area of the section as well as the length of the trachealis muscle were significantly greater; although the thickness of the muscle and cartilage were seen to be significantly lower in group II sections compared to group I sections. Also, in comparison to group I, in group II sections there was lesser overlap of the posterior free ends of tracheal cartilage and the epithelial layer was flattened and focally abraded. Our findings demonstrate structural changes in the airway of preterm animals and characterize alterations in the geometric arrangement of muscle and cartilage after PPV. These results suggest possible structural mechanisms for the functional changes seen during and subsequent to mechanical ventilation.

Animals

Long-term structural changes in the rat hippocampal formation following cerebral ischemia.

Histological and immunohistochemical techniques were used to investigate the long-term structural changes that occur in the rat hippocampal formation following the induction of transient forebrain ischemia. Histological analysis showed that after 6-12 months cell loss was still largely restricted to the CA1 region but within this region degeneration was progressive and culminated in a severe shrinkage of the stratum oriens and stratum radiatum. Using the immunohistochemical markers calbindin-D28K and parvalbumin, we were able to demonstrate some of the structural changes that reflect this shrinkage. Calbindin immunohistochemistry clearly illustrated that the shrinkage was primarily due to the loss of the pyramidal neurons and the framework normally provided by their long apical dendrites. Parvalbumin immunostaining demonstrated that although a few GABAergic interneurons survived the insult, their terminal network was reduced greatly and the dendrites which normally extended the length of the stratum radiatum were retracted. Additionally, the normally dense band of parvalbumin immunoreactivity in the stratum lacunosum-moleculare, thought to be indicative of a fiber bundle travelling through the CA1 region, was almost completely depleted. These data illustrate that the primary damage observed in the CA1 region following cerebral ischemia is not static but progressive and may thus have important functional implications.

Animals

Time-resolved X-ray diffraction study of structural changes associated with the photocycle of bacteriorhodopsin.

The time course of structural changes accompanying the transition from the M412 intermediate to the BR568 ground state in the photocycle of bacteriorhodopsin (BR) from Halobacterium halobium was studied at room temperature with a time resolution of 15 ms using synchrotron radiation X-ray diffraction. The M412 decay rate was slowed down by employing mutated BR Asp96Asn in purple membranes at two different pH-values. The observed light-induced intensity changes of in-plane X-ray reflections were fully reversible. For the mutated BR at neutral pH the kinetics of the structural alterations (tau 1/2 = 125 ms) were very similar to those of the optical changes characterizing the M412 decay, whereas at pH 9.6 the structural relaxation (tau 1/2 = 3 s) slightly lagged behind the absorbance changes at 410 nm. The overall X-ray intensity change between the M412 intermediate and the ground state was about 9% for the different samples investigated and is associated with electron density changes close to helix G, B and E. Similar changes (tau 1/2 = 1.3-3.6 s), which also confirm earlier neutron scattering results on the BR568 and M412 intermediates trapped at -180 degrees C, were observed with wild type BR retarded by 2 M guanidine hydrochloride (pH 9.4). The results unequivocally prove that the tertiary structure of BR changes during the photocycle.

Bacteriorhodopsins

[Structural changes in the kidneys in myocardial infarct].

The structural changes of kidneys in myocardial infarction patients were investigated by histological, histochemical and histometric methods. Results indicate that the kidneys of these patients showed dystrophic, necrobiotic and necrotic changes related to disorders of both general and intrarenal hemodynamics.

Aged

[Structural changes in ganglioside micelles during interaction with serotonin].

Structure changes of ganglioside micelles in the process of serotonin bonding are considered. It was shown by viscosimetric and differential refraction studies that stretching of elliptical ganglioside micelles in the serotonin bonding process took place. The scope of micelles form changes depends on the pH value. An increase of the micelles summary negative charge at pH 9 increased the number of bonding centres and simultaneously caused deeper deformation of micelles due to the repulsion of the identically charged groups.

Binding Sites

Structural changes in the cell wall of Schizosaccharomyces pombe during cell division.

Streiblová, Eva (Czechoslovak Academy of Sciences, Prague, Czechoslovakia), I. Málek, and K. Beran. Structural changes in the cell wall of Schizosaccharomyces pombe during cell division. J. Bacteriol. 91:428-435. 1966.-Individual stages of growing and dividing cells of Schizosaccharomyces pombe were studied by means of fluorescence and electron microscopy with the use of metal-shadowed isolated walls, replicas, and ultrathin sections. Vegetative cells were found to contain division scars (six at the most); their formation and structure are described. More data on the growth of arthrospores were obtained. New structural observations were made on the architecture of the cell wall (original wall ring, polar cell wall, plug wall band, additional wall ring). Structural changes of cell surfaces and lateral walls during fission are represented schematically to the fourth generation. The question of origin of the septum is discussed, and on this basis the entire structure of the cell wall is interpreted.

Ascomycota

Crystal structures of modified myoglobins. I. Heme orientation and structural changes around heme in myoglobins reconstituted with isopemptoheme, pemptoheme, 2-ethyldeuteroheme, and 4-ethyldeuteroheme.

The crystal structures of sperm whale metmyoglobins reconstituted with four modified hemes, isopemptoheme, pemptoheme, 2-ethyldeuteroheme, and 4-ethyldeuteroheme, have been determined and refined at 2.2 A resolution to R = 0.217, 0.218, 0.213, and 0.222, respectively. All the crystals of these myoglobins are isomorphous with that of native metmyoglobin. The structural changes of the modified myoglobin from the native myoglobin were examined on difference Fourier maps; the orientation of 4-ethyldeuteroheme in the heme pocket is such that the heme is rotated by 180 degrees about an axis through the alpha-gamma-meso carbons, whereas the orientations of the other three hemes are the same as that of the protoheme in the native myoglobin. The changes of the structures around the heme become greater in the order of isopemptoheme, 2-ethyldeuteroheme less than pemptoheme less than 4-ethyldeuteroheme. The magnitudes of the changes seem to be related to the oxygen affinities of these four reconstituted myoglobins.

Animals