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The ultrastructure and vascular supply of the different fibre types in the axial muscle of the sturgeon Acipenser stellatus, Pallas.

The ultrastructure and vascular supply of the different fibre types in the lateral muscles of the sturgeon Acipenser stellatus were studied by light- and electron microscopy and morphometry. Three fibre types form separate layers without intermingling. The red fibres are superficial, the white fibres deep and the intermediate fibres between them. From morphometric analyses, the mitochondrial volume fraction in red fibres is 30%, in intermediate fibres 3.7% and in white fibres 0.7%. Z lines are most fuzzy in the red fibres. Triads of the sarcotubular system are always situated at the level of the Z discs. In red fibres the three elements are arranged in a series along the myofibrils, whereas in white fibres they are arranged transversely and in the intermediate fibres they are aligned obliquely. The number of capillaries surrounding each fibre is 2.3, 0.9 and 0.2 for the red, intermediate and white fibres, respectively. In red fibres 16% of the surface is directly covered by capillaries. The corresponding percentages for intermediate and white fibres are 5 and 1, respectively. Per unit volume of the fibre, the directly vascularised fibre surface in red fibres is about ten times larger than that of white fibres. The degree of vascularisation of the fibre types is directly related to the volume fraction of mitochondria, and thus to their aerobic capacities.

Animals↗

Validation of a computer analysis to determine 3-D rotations and translations of the rib cage in upright posture from three 2-D digital images.

Since thoracic cage posture affects lumbar spine coupling and loads on the spinal tissues and extremities, a scientific analysis of upright posture is needed. Common posture analyzers measure human posture as displacements from a plumb line, while the PosturePrint claims to measure head, rib cage, and pelvic postures as rotations and translations. In this study, it was decided to evaluate the validity of the PosturePrint Internet computer system's analysis of thoracic cage postures. In a university biomechanics laboratory, photographs of a mannequin thoracic cage were obtained in different postures on a stand in front of a digital camera. For each mannequin posture, three photographs were obtained (left lateral, right lateral, and AP). The mannequin thoracic cage was placed in 68 different single and combined postures (requiring 204 photographs) in five degrees of freedom: lateral translation (Tx), lateral flexion (Rz), axial rotation (Ry), flexion-extension (Rx), and anterior-posterior translation (Tz). The PosturePrint system requires 13 reflective markers to be placed on the subject (mannequin) during photography and 16 additional "click-on" markers via computer mouse before a set of three photographs is analyzed by the PosturePrint computer system over the Internet. Errors were the differences between the positioned mannequin and the calculated positions from the computer system. Average absolute value errors were obtained by comparing the exact inputted posture to the PosturePrint computed values. Mean and standard deviation of computational errors for sagittal displacements of the thoracic cage were Rx=0.3+/-0.1 degrees , Tz=1.6+/-0.7 mm, and for frontal view displacements were Ry=1.2+/-1.0 degrees , Rz=0.6+/-0.4 degrees , and Tx=1.5+/-0.6 mm. The PosturePrint system is sufficiently accurate in measuring thoracic cage postures in five degrees of freedom on a mannequin indicating the need for a further study on human subjects.

Biomechanical Phenomena↗

Cell lineage analysis in ascidian embryos by intracellular injection of a tracer enzyme. II. The 16- and 32-cell stages.

Cell lineages during development of ascidian embryos were analyzed by injecting horseradish peroxidase as a tracer enzyme into identified cells of the 16-cell and 32-cell stage embryos of Halocynthia roretzi. Most of the blastomeres of these embryos developed more kinds of tissues than have hitherto been reported, and therefore, the developmental fates of each blastomere are more complex. It has been thought that every blastomere of the 64-cell stage ascidian embryo gives rise to only one kind of tissues, but the finding that the several blastomeres at the 32-cell stage developed into at least three different kinds of tissues, clearly indicates that the stage at which the fates of every blastomere are determined to one tissue is later than the 64-cell stage. The results also clearly demonstrate that muscle cells are derived not only from B-line cells (B5.1, B5.2, B6.3, and B6.4) but also from A-line cells (A5.2 and A6.4) and b-line cells (b5.3 and b6.5). Based on the present analysis as well as other studies, complete cell lineages of muscle cells up to their terminal differentiation have been proposed. In addition, lineages of nervous system, notochord, and epidermis are also discussed.

Animals↗

The coupling ATPase complex: an evolutionary view.

Phospholipid micelles and vesicles, present in the primordial soup, formed both primitive (surface) catalyst and primitive replicative life forms. With the adoption of a common energy source, ATP, integrated biochemical systems within these vesicles became possible - cells. Fermentation within these primitive cells was favoured by the evolution, first of ion channels allowing protons to leak out, and then of an active ATP-driven pump. In the prokaryotic/mitochondria/chloroplast line, the proton channel was such as to be blocked by dicyclohexylcarbodiimide and the adenosine 5' triphosphate phosphohydrolase (ATPase) by 4-chloro 7-nitrobenzofurazan (Nbf-C1). The ATPase was initially simple (4 subunits) but later, possibly concomitant with its evolution to an ATP synthetase, became more complex (8 subunits). One of the steps in evolution probably involved gene duplication and divergence of 2 subunits (alpha and beta) from the largest of the ATPase subunits. From this stage, the general form of the ATPase was fixed, although sensitivity to, for example, oligomycin involved later, after divergence of the mitochondrial and chloroplast lines. A regulatory protein, the ATPase inhibitor, is found associated with a wide spectrum of coupling ATPases.

Adenosine Triphosphatases↗

Lateral organization and domain formation in a two-component lipid membrane system.

The thermodynamic phase behavior and lateral lipid membrane organization of unilamellar vesicles made from mixtures of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and 1,2 distearoyl-sn-glycero-3-phosphocholine (DSPC) were investigated by fluorescence resonance energy transfer (FRET) as a function of temperature and composition. This was done by incorporating a headgroup-labeled lipid donor (NBD-DPPE) and acceptor (N-Rh-DPPE) in low concentrations into the binary mixtures. Two instances of increased energy transfer efficiency were observed close to the phase lines in the DMPC/DSPC phase diagram. The increase in energy transfer efficiency was attributed to a differential preference of the probes for dynamic and fluctuating gel/fluid coexisting phases. This differential preference causes the probes to segregate (S. Pedersen, K. Jørgensen, T. R. Baekmark, and O. G. Mouritsen, 1996, Biophys. J. 71:554-560). The observed increases in energy transfer match with the boundaries of the DMPC/DSPC phase diagram, as measured by Fourier transform infrared spectroscopy (FTIR) and differential scanning calorimetry (DSC). We propose that the two instances of probe segregation are due to the presence of DMPC-rich and DSPC-rich domains, which form a dynamic structure of gel/fluid coexisting phases at two different temperatures. Monitoring the melting profile of each lipid component independently by FTIR shows that the domain structure is formed by DMPC-rich and DSPC-rich domains rather than by pure DMPC and DSPC domains.

Calorimetry, Differential Scanning↗

Modulation of expression of intermediate filaments during the development of established rat liver cell lines.

A number of clear epithelial-like cell lines were established from the liver of fetal and neonatal rats. The intermediate filaments of these cells were investigated using polyclonal prekeratin antisera, monoclonal antibodies against a cytokeratin subfamily and vimentin by means of the indirect immunofluorescence technique and SDS-PAGE analysis. Changes in the expression of cytokeratin filaments were found during the evolution of permanent cell lines. Cells positively stained for cytokeratins could be seen near to other cells which were negative. In early passages (up to the 40th) nearly all cells were strongly stained by the different keratin antibodies. During the following subcultivation the pattern of staining considerably changed. In FRL and NP-RL cell lines keratin-negative cells could already be observed in the early passages, rapidly increasing in the later passages. Compared to this, vimentin-staining of all cells remained constant in its morphological expression. The keratin filaments were seen in thick fiber bundles arranged particularly in the perinuclear ring as well as in finer networks throughout the cytoplasm. Every cell in the established lines showed their very individual staining pattern. The vimentin filaments extended to the whole cytoplasm up to the cell margin. Our observations demonstrate the variability of the system of keratin filaments in established epithelial-like liver cells under cultural conditions.

Animals↗

Facilitatory interactions between flashed lines.

The detectability of pairs of flashed lines was investigated as a joint function of their separation in both space and time. In contrast to previous studies of contrast interactions as a function of either spatial or temporal separation alone, the predominant interaction is not lateral inhibition, but a delayed facilitation. A quantitative model based on probability summation between lateral-inhibitory detectors does not appear able to account for these results. Two related explanations are suggested: either a motion detection system or the nonlinear 'transient' subsystem is being tapped.

Humans↗

[The treatment of iatrogenic pneumothorax with small-gauge catheters. The author's personal experience in 30 cases].

PURPOSE: Pneumothorax (PNX) is the most frequent complication in patients who have undergone lung biopsy. If PNX is asymptomatic and < 30%, it does not require treatment, while if it is > 30% and the patient is symptomatic treatment is needed. As a rule surgery is required and patients are hospitalized and undergo intrathoracic drainage with positioning of a large gauge catheter--i.e. over 15 French (F). In the last 10 years radiologists have begun treating PNX with much smaller catheters (7-10 F). We report the execution technique using 6.3 F catheters and the results obtained in 30 patients with symptomatic iatrogenic PNX and/or iatrogenic PNX > 30%. MATERIAL AND METHODS: All the patients underwent CT-guided lung biopsy. Immediately after the procedure some follow-up scans were performed and a further expiratory radiograph with the patient in upright position was carried out after at least 2 hours. If an asymptomatic PNX < 30% was found the patient was discharged and submitted to radiographic follow-up the following morning and every 24 hours thereafter for 2 days. If there was a symptomatic PNX and/or a PNX > 30% an intrathoracic drainage catheter was positioned. Under fluoroscopic or CT guidance we positioned a 5.7 F intrathoracic pig-tail catheter at a point corresponding to the 3rd or 4th intercostal space on the midclavear line. After manual suction of intrathoracic air we connected the catheter to a Hemlick valve and repeated the chest radiograph 4 hours later. If the PNX had not reformed the patient was discharged and submitted to radiographic follow-up every 24 hours for 3-5 days. On the contrary if the PNX had reformed, or if pain and/or dyspnea symptoms or signs persisted, the catheter was connected to a continuous-suction system and the patient rehospitalized for about 6 days. Oximetry was performed in all patients before biopsy, on PNX diagnosis, and after pulmonary re-expansion. RESULTS: All the cases were resolved and 9 patients were followed-up in the outpatients department. Drainage had to be repeated in 2 patients only and the 5.7 F catheters replaced with an 8 F and a 10 F catheters. Oximetric data were always correlated with the presence/absence of PNX. In particular, in PNX > 30% we found over 10% reduction relative to prebiopsy values. This datum was corrected and came to meet the prebiopsy value as soon as the lung was re-expanded. No significant changes were seen in PNX < 30%. CONCLUSIONS: Small gauge catheters provide the following advantages: the procedure presents a low risk of complications, is easy to carry out and much better tolerated by the patient; also in some cases the cost is lower because no hospitalization is required. The close correlation of oximetric values with the presence/absence of PNX < 30% could be considered to decrease follow-up radiographic examinations. Finally the possibility of treating iatrogenic PNX using radiological techniques further promotes the acceptability of lung biopsy by colleagues from other branches of medicine.

Biopsy, Needle↗

Electron paramagnetic resonance imaging of nitric oxide organ distribution in lipopolysuccaride treated mice.

The recent development of electron paramagnetic resonance (EPR) permits its application for in vivo studies of nitric oxide (NO). In this study, we tried to obtain 3D EPR images of endogenous NO in the abdominal organs of lipopolysuccaride (LPS) treated mice. Male ICR mice, each weighing about 30 g, received 10 mg/kg of LPS intraperitoneally. Six hours later, a spin trapping reagent comprised of iron and an N-dithiocarboxy sarcosine complex (Fe(DTCS)2, Fe 200 mM, DTCS/Fe = 3) were injected subcutaneously. Two hours after this treatment, the mice were fixed in a plastic holder and set in the EPR system, equipped with a loop-gap resonator and a 1 GHz microwave. NO was detected as an NO-Fe(DTCS)2 complex, which had a characteristic 3-line EPR spectrum. NO-Fe(DTCS)2 complexes in organ homogenates were also measured using a conventional X-band EPR system. NO-Fe(DTCS)2 spectra were obtained in the upper abdominal area of LPS treated mice at 8 h after the LPS injection. 3D EPR tiled and stereoscopic images of the NO distribution in the hepatic and renal areas were obtained at the same time. The NO-Fe(DTCS)2 distribution in abdominal organs was confirmed in each organ homogenate using conventional X-band EPR. This is the first known EPR image of NO in live mice kidneys.

Animals↗

[Changes in the distribution of adenylate cyclase activity in PC 12 cells under the influence of nerve growth factor].

Changes in distribution of adenylate cyclase in PC 12 cells under the influence of nerve growth factor (NGF) have been studied using cytochemical methods. The adenylate cyclase activity was predominantly associated with the plasma membrane. In cell cultures without NGF the activity was revealed on the contacting surfaces of cell aggregates; single grains of reaction product were revealed on exposed cell surface only in cultures with a high cell density. One day after administration of NGF, the adenylate cyclase activity on exposed cell surface increased, and three days later the whole cell surface was covered with lead sediment. The enzyme activity was also revealed in growth cones, filopodia and microcytospheres. The role of adenydlate cyclase system in neuron-like differentiation of PC 12 cells is discussed.

Adenylyl Cyclases↗

Air embolism in central venous catheterization: diagnosis, treatment, and prevention.

Air embolism associated with central venous catheterization carries with it a significant morbidity and mortality. The diagnosis should be suggested by sudden alterations in cardiovascular, respiratory, or central nervous system function in a patient with a central venous catheter. A "mill wheel" cardiac murmur is characteristic. Placing the patient in the left lateral decubitus with the head down allows displacement of the air from the pulmonary outflow tract. Prevention of the complication involves thorough patient and physician preparation before subclavian catheterization, use of a Luer lock between catheter and tubing, meticulous catheter care, and employment of a pump with an in-line air detector.

Adult↗

Integrin alpha 6 expression is required for early nervous system development in Xenopus laevis.

The integrin alpha 6 subunit pairs with both the beta 1 and beta 4 subunits to form a subfamily of laminin receptors. Here we report the cDNA cloning and primary sequence for the Xenopus homologue of the mammalian integrin alpha 6 subunit. We present data demonstrating the spatial and temporal expression of alpha 6 mRNA and protein during early development. Initially, alpha 6 transcripts are expressed in the dorsal ectoderm and future neural plate at the end of gastrulation. Later in development, alpha 6 mRNAs are expressed in a variety of neural derivatives, including the developing sensory placodes (otic and olfactory) and commissural neurons within the neural tube. Integrin alpha 6 is also expressed in the elongating pronephric duct as well as a subset of the rhombencephalic neural crest, which will form the Schwann cells lining several cranial nerves (VII, VIII and X). In vivo expression of an alpha 6 antisense transcript in the animal hemisphere leads to a reduction in alpha 6 protein expression, a loss of adhesion to laminin, and severe defects in normal development. In 35% of cases, reduced levels of alpha 6 expression result in embryos that complete gastrulation normally but arrest at neurulation prior to the formation of the neural plate. In an additional 22% of cases, embryos develop with severe axial defects, including complete loss of head or tail structures. In contrast, overexpression of the alpha 6 subunit by injection of full-length mRNA has no apparent effect on embryonic development. Co-injection of antisense and sense plasmid constructs results in a partial rescue of the antisense-generated phenotypes. These data indicate that the integrin alpha 6 subunit is critical for the early development of the nervous system in amphibians.

3T3 Cells↗

A tibial-based coordinate system for three-dimensional data.

An accurate and repeatable tibial measurement system will aid in the definition of tibial geometry and improving tibial prosthesis design. Unlike in the femur, there is no standardized method for constructing a tibial coordinate frame. Most tibial measurements are given relative to femoral axes or the coordinate frame of the CT/MRI scanner or radiograph machine. The objective of this study was to establish an independent tibial coordinate frame. Data consisted of CT scans from 34 subjects. The tibial anatomical axis was chosen as the axial axis. The anteroposterior (AP) axis was selected to be parallel to the lateral surface of the tibial shaft and orthogonal to the anatomical axis and from this the mediolateral axis could be derived. The selected AP axis was compared with the surgical tibial AP axis by measuring their variability relative to a common axis, the posterior tibial condylar line (PTCL). The mean angle between the selected AP axis and the perpendicular to the PTCL was measured as -4.07 degrees, standard deviation of 4.28 degrees. The mean angle between the surgical AP axis and the perpendicular to the PTCL was measured as -18.56 degrees, standard deviation of 4.66 degrees. There was no significant difference in the variance of the two sets of measurements (p=0.63). Variability of the selected AP axis was even smaller (standard deviation of 2.74 degrees) when measured independently from the PTCL reference axis, by aligning virtual resection profiles. Anatomically, the selected AP axis was almost perpendicular to the posterior tibial condylar axis. This coordinate system can aid in gathering consistent and repeatable anthropometric data that can be used to improve tibial implant design and could also, in combination with CT/MR imaged-based computer assisted surgery, be used as a guideline for tibial component positioning in TKR.

Adult↗

A system for monocytic differentiation of leukemic cells HL 60 by a short exposure to 1,25-dihydroxycholecalciferol.

The human promyelocytic cell line HL 60 can be induced to differentiate toward more mature myeloid or monocytic forms by a variety of agents. This process is thought to require several days of exposure to the inducer, thus making it difficult to identify the early cellular changes which are fundamental to the differentiation program, and to relate the induction to phases of the cell cycle. In order to study the kinetics of leukemic cell differentiation we have developed a system for the induction of rapid monocytic maturation in a subpopulation of HL 60 cells. The cells are exposed to 10(-7) M 1,25-dihydroxycholecalciferol for 4 hr in serum-free medium. Subsequent incubation in a complete medium results in cellular differentiation recognizable by several criteria (phagocytosis, nonspecific esterase reaction, adherence to substratum, cell morphology) beginning at 10 hr from the exposure to the inducer. Approximately 20 hr later 30-40% of the cells in culture show the differentiated phenotype and are capable of phagocytosis. The proportion of differentiated cells in culture decreases thereafter. This system has been utilized to study the expression of c-myc oncogene in relation to the kinetics of maturation, and it was found that the inhibition of the expression of this gene precedes the onset of phenotypic differentiation by approximately 8 hr, is transient, and is accompanied by a brief retardation of cell proliferation, which resumes the normal rate within 24 hr of the exposure to the inducer.

Calcitriol↗

Human ocular counterrolling induced by varying linear accelerations.

Ocular counterrolling (OCR) has previously been studied using static head tilt or continuous rotation about the line of sight as a stimulus to the otolith organs. This study presents the first measurements of OCR in humans induced by linear accelerations. Dynamic measurements of the response to lateral linear acceleration indicate the eye movements to be on the order of 2 degrees for 0.2 g peak acceleration, 0.2 Hz sinusoidal acceleration. These values are consistent with static OCR studies. The dynamics of the response are similar to a low order linear system with a dominant time constant of 0.33 s. A previous model predicts a time constant of 0.32 s. Sinusoidal oscillation at 0.2, 0.4, and 1.0 Hz with a 0.2 g peak acceleration showed good agreement with the model in both gain and phase. The question of amplitude linearity remains unsettled. This otolithocular reflex, over short periods at least, appears to be stationary in the statistical sense.

Acceleration↗

Nitric oxide synthase in the glossopharyngeal and vagal afferent pathway of a teleost, Takifugu niphobles. The branchial vascular innervation.

To examine the presence of nitric oxide synthase (NOS) in the sensory system of the glossopharyngeal and vagus nerves of teleosts, nicotinamide adenine dinucleotide phosphate diaphorase (NADPHd) activity and immunoreactivity for NOS were examined in the puffer fish Takifugu niphobles. The nitrergic sensory neurons were located in the ganglia of both the glossopharyngeal and the vagal nerves. In the vagal ganglion, positive neurons were found in the subpopulations for the branchial rami and the coelomic visceral ramus, but not for the posterior ramus or the lateral line ramus. In the medulla, nitrergic afferent terminals were found in the glossopharyngeal lobe, the vagal lobe, and the commissural nucleus. In the gill structure, the nitrergic nerve fibers were seen in the nerve bundles running along the efferent branchial artery of all three gill arches. These fibers appeared to terminate in the proximal portion of the efferent filament arteries of three gill arches. On the other hand, autonomic neurons innervating the gill arches were unstained. These results suggest that nitrergic sensory neurons in the glossopharyngeal and vagal ganglia project their peripheral processes through the branchial rami to a specific portion of the branchial arteries, and they might play a role in baroreception of this fish. A possible role for nitric oxide (NO) in baroreception is also discussed.

Afferent Pathways↗

Myocardial infarct expansion: recognition, significance and pathology.

Infarct expansion can be defined pathologically as a distortion of ventricular topography produced by thinning and disproportionate dilation of the infarct segment. Large transmural infarcts tend to be associated with greater propensity for infarct expansion. Two-dimensional echocardiography has made it feasible to detect these acute alterations in cardiac topography by serial examination of patients with acute myocardial infarction. A practical approach to the echocardiographic quantification of expansion involves analysis of end-diastolic cross-sectional echo views at the papillary muscle level, which can be used as fixed internal landmarks to divide the left ventricle into 2 segments, anterior and posterior. An off-line computer system can be used to track relative lengths of these segments as well as their thicknesses over time. In the initial clinical study, one third of patients with acute anterior transmural infarcts showed an average 50% increase in the infarct segment length beginning within the first 3 days of infarction, characterized by disproportionate progressive dilation and transmural thinning of this zone. These patients demonstrated a significantly higher mortality than those without expansion. Later studies demonstrated not only continuing dilation of the infarcted anterior wall, but also progressive dilation of the noninfarcted posterior wall, underscoring the importance of continuing long-term noninvasive follow-up. Not only is expansion associated with a poor clinical outcome; it has also been shown experimentally and clinically to be modifiable or even preventable by various therapeutic maneuvers, which may well improve survival. Because of the limitations of the echocardiographic window, it is often possible to obtain only a single cross-sectional view of high quality, and even then technical quality may not be sufficiently high to enable detailed quantitative analysis.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Influence of cholesterol on gramicidin-induced HII phase formation in phosphatidylcholine model membranes.

The influence of cholesterol incorporation on gramicidin-induced hexagonal HII phase formation in different phosphatidylcholine model systems was investigated by 31P- and 2H-NMR, small-angle X-ray diffraction and differential scanning calorimetry. In liquid-crystalline distearoylphosphatidylcholine systems cholesterol inhibits gramicidin-induced HII phase formation. In dioleoylphosphatidylcholine the opposite effect is observed. Cholesterol appears to preferentially interact with gramicidin under liquid-crystalline conditions in both systems. Two phenomena that had been reported for gramicidin-treated erythrocyte membranes and derived liposomes (Tournois, H., Leunissen-Bijvelt, J., Haest, C.W.M., De Gier, J. and De Kruijff, B. (1987) Biochemistry, 26, 6613-6621) could also be observed in more simple dioleoylphosphatidylcholine-gramicidin-cholesterol systems. These are (i) an increase in tube diameter in the gramicidin-induced HII phase with increasing temperature, which is ascribed to the presence of cholesterol in this phase, and (ii) the loss of the hexagonal HII phase related 31P-NMR line shape at lower temperatures despite the presence of this phase as demonstrated with X-ray diffraction. This latter phenomenon appears to be due to restrictions in the rate of lateral diffusion of the phospholipids around the HII tubes due to the presence of gramicidin.

Chemical Phenomena↗