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PubMed1985

Model of gas transport during high-frequency ventilation.

We analyze gas exchange during high-frequency ventilation (HFV) by a stochastic model that divides the dead space into N compartments in series where each compartment has a volume equal to tidal volume (V). We then divide each of these compartments into alpha subcompartments in series, where each subcompartment receives a well-mixed concentration from one compartment and passes a well-mixed concentration to another in the direction of flow. The number of subcompartments is chosen on the basis that 1/alpha = (sigma t/-t)2, where -t is mean transit time across a compartment of volume, and sigma t is standard deviation of transit times. If (sigma t/-t)D applies to the transit times of the entire dead space, the magnitude of gas exchange is proportional to (sigma t/-t)D, frequency, and V raised to some power greater than unity in the range where V is close to VD. When V is very small in relation to VD, gas exchange is proportional to (sigma t/-t)2D, frequency, and V raised to a power equal to either one or two depending on whether the flow is turbulent or streamline, respectively. (sigma t/-t)D can be determined by the relation between the concentration of alveolar gas at the air outlet and volume expired as in a Fowler measurement of the volume of the dead space.

Humans↗
PubMed2003

A multicompartment model of carboxyhemoglobin and carboxymyoglobin responses to inhalation of carbon monoxide.

We have developed a model that predicts the distribution of carbon monoxide (CO) in the body resulting from acute inhalation exposures to CO. The model includes a lung compartment, arterial and venous blood compartments, and muscle and nonmuscle soft tissues with both vascular and nonvascular subcompartments. In the model, CO is allowed to diffuse between the vascular and nonvascular subcompartments of the tissues and to combine with myoglobin in the nonvascular subcompartment of muscle tissue. The oxyhemoglobin dissociation curve is represented by a modified Hill equation whose parameters are functions of the carboxyhemoglobin (HbCO) level. Values for skeletal muscle mass and cardiac output are calculated from prediction formulas based on age, weight, and height of individual subjects. We demonstrate that the model fits data from CO rebreathing studies when diffusion of CO into the muscle compartment is considered. The model also fits responses of HbCO to single or multiple exposures to CO lasting for a few minutes each. In addition, the model reproduces reported differences between arterial and venous HbCO levels and replicates predictions from the Coburn-Forster-Kane equation for CO exposures of a 1- to 83-h duration. In contrast to approaches based on the Coburn-Forster-Kane equation, the present model predicts uptake and distribution of CO in both vascular and tissue compartments during inhalation of either constant or variable levels of CO.

Adult↗
PubMed2000

Ribosome biogenesis in man: current views on nucleolar structures and function.

Nucleoli develop when preribosomes are synthesized at the chromosomal nucleolar organizer regions. Typically they consist of at least three nucleolar subcompartments, the fibrillar center (FC), the dense fibrillar component (DF), and the granular component (GC). The understanding of the functional arrangements of these subcompartments relates to aspects in cell biology, pathology, and virus research. In the present review morphological studies are discussed in the light of molecular findings. The available data confirm the hypothesis that rDNA transcription is connected with the DF but not necessarily with the presence of an FC. Within the DF, rDNA transcription is restricted to foci, possibly representing single transcribing genes. FCs may serve to store inactive transcription factors, to initiate rDNA transcription, and may provide structural support for transcription. The GC can be interpreted as a collection of maturing preribosomes. More recently the nucleolar subcompartments were focused on in the context of virus research and tumor biology.

Cell Nucleolus↗
PubMedDate not supplied

Chronic exertional compartment syndrome: the controversial "fifth" compartment of the leg.

BACKGROUND: The existence of a "fifth" compartment in the leg capable of developing distinct chronic exertional compartment syndrome remains a subject of controversy. HYPOTHESIS: Specific pressure recordings and dissection will confirm or disprove the existence of a fifth compartment. STUDY DESIGN: Empirical anatomic study. METHODS: Radiopaque dye was injected directly into the tibialis posterior muscle of 25 embalmed cadaveric legs while intracompartmental pressure was monitored. Radiographs demonstrated dye distribution, and dissection-documented fascial and epimysial layers. RESULTS: Evidence was found that the fibular origin of the flexor digitorum longus muscle, when present, could create subcompartments within the deep posterior compartment. The nature of this attachment varied from being absent, to small (<8 cm), to extensive (>8 cm). The attachment partially covered the tibialis posterior muscle in the majority of the 14 legs that developed high pressures, and it was limited or absent in the 11 legs that did not. Radiographs demonstrated that the dye was confined to the tibialis posterior muscle in four legs. CONCLUSIONS: No consistent fifth compartment exists in the leg; however, subcompartments within the deep posterior compartment created by the fibular origin of the flexor digitorum longus muscle may develop pressures congruent with chronic exertional compartment syndrome. CLINICAL RELEVANCE: Potential deep posterior subcompartments demand accurate pressure investigation. A modified technique to decompress the entire deep posterior compartment, including the tibialis posterior muscle, is necessary for successful treatment of chronic exertional compartment syndrome.

Aged↗
PubMed1998

Site-selective differences in cytochrome P450 isoform activities. Comparison of expression in rat and rhesus monkey lung and induction in rats.

The distribution of pulmonary cytochrome P450 (P450 or CYP) isoforms has been investigated primarily in immunohistochemical studies, which are neither quantitative nor reflective of the functions of these enzymes. Studies of enzyme activities have been performed using whole-lung homogenates or isolated cells, but there is little information on the regioselective expression of P450 monooxygenases. The aims of this study were to compare the activities of P450 monooxygenases in different lung subcompartments in two commonly studied animal models, i.e. rats and monkeys, and to explore the possibility that inducing agents would result in activity up-regulation that is highly site-selective, using rats as a model. Microdissection techniques were used to separate the airways from blood vessels and lung parenchyma. In rats, CYP1A1 (ethoxyresorufin) and CYP2B (pentoxyresorufin) dealkylase activities were highest in the parenchyma, whereas CYP2E1 (p-nitrophenol) hydroxylase activity was highest in the airways. P450 reductase activities were similar in airways and parenchyma and were lower in trachea. In monkeys, no significant site-selective differences in CYP1A1 and CYP2B1 activities were found. In contrast, CYP2E1 activity was higher in the distal bronchioles and parenchyma than in the proximal airways. P450 reductase activities were similar in microsomes prepared from all subcompartments of monkey lung. Induction of rat CYP1A1 activity by beta-naphthoflavone (administered ip) was much greater in the airways and lung parenchyma ( approximately 30-fold) than in the liver ( approximately 10-fold) or trachea ( approximately 2.5-fold). Oral administration of phenobarbital or acetone increased CYP2B and CYP2E1 activities in rat liver but had no significant effect on P450 activities in subcompartments of rat lung. These findings support the conclusion that there are regiospecific and species-specific differences in the activities of P450 isoforms and that the inducibility of rat pulmonary P450s is dependent on the isoform and lung region.

Animals↗
PubMed1991

Organization of the reticular network of rabbit Peyer's patches.

The organization of the network of collagen fibers of rabbit Peyer's patches was examined by scanning electron microscopy (SEM) in alkali-water macerated tissues. The relationship between this network and the reticular cells within it was further studied by SEM of ultrasonicated tissues. Collagen fibrils (about 60 nm in diameter) formed collagen fibers or sheets. There were sheets of collagen fibrils with numerous pores beneath the patch dome epithelium. Within the patches, collagen fibers repeatedly divided and fused, forming the reticular network. The reticular network within the follicle was looser than within the dome, the corona, or the interfollicular area. The latter three compartments showed similar structures and consisted of numerous intercommunicating small subcompartments. Reticular cells were in contact with groups of free cells lodged in these subcompartments within the reticular network. Reticular cell processes with numerous fenestrations embraced not only collage fibers forming the reticular network, but also sheaths of collagen fibers of blood and lymphatic vessels. Sheaths of collagen fibers of high endothelial venules and lymphatic vessels were also fenestrated, indicating the sites through which lymphocytes and other free cells migrate. These results indicate that the reticular network of Peyer's patches is organized so as to facilitate migration and lodging of free cells and thus facilitate antigen-to-cell and cell-to-cell interactions during an immune response. The naked areas on the collagen fibers seem to provide a scaffolding for free cells during their migration.

Animals↗
PubMed1992

Changes of cytochemical properties in the Golgi apparatus during in vivo differentiation of the ameloblast in developing rat molar tooth germs.

The cytochemical changes of the Golgi stacks occurring concomitantly with cell differentiation were examined in ameloblasts of developing rat molar tooth germs using osmium impregnation and cytochemistry with nicotinamide adenine dinucleotide phosphatase (NADPase), thiamine pyrophosphatase (TPPase), and acid phosphatase (Acpase). NADPase, TPPase, and Acpase activities were already present in the Golgi stacks of the inner enamel epithelial cells, the undifferentiated form of the ameloblast: NADPase activity existed in the medial Golgi cisternae, TPPase activity in the trans Golgi cisternae, and Acpase activity in almost all cisternae and strongly in the trans-most cisterna of the Golgi stack. At this stage, however, osmium deposits after impregnation were not observed in the cisterna of Golgi stacks but were present in some small vesicles. These vesicles were located throughout the cytoplasm. Osmiophilic cisternae in the Golgi stacks were apparent for the first time at the stage when the Golgi apparatus developed and migrated to the region distal to the nucleus with the progression of cell differentiation. These findings indicate that the cis subcompartment of the Golgi apparatus was incomplete in the inner enamel epithelial cells with regard to appearance of its cytochemical property, as compared with the medial and trans subcompartments. It is suggested that the cis compartment of the Golgi stack may be completed only in the last stage of the compartmentalized Golgi organization during differentiation of the ameloblast.

Acid Phosphatase↗
PubMed1997

A multicompartment geometric model of the liver in relation to regional induction of cytochrome P450s.

A geometric, multicompartment model of the liver was developed to examine regional protein induction and to provide model output suitable for predicting the degree of induction in both the whole liver and in specific regions. The model was based on functional hexagonal arrays within the liver. A geometric representation was used to divide these functional units into five zones: a concentric periportal zone, a fenestrated periportal region that interconnects among multiple functional units, and three concentric centrilobular areas, referred to, respectively, as zones 1 through 5. The surface areas (and volumes for hexagonal cylinders) of these live zones were, respectively, 13.5, 25.2, 33.9, 20.3, and 6.8% of the total liver. The pharmacokinetic model for induction had dissociation constants (Kd) and Hill constants (n) for interactions of transcriptional activator-ligand complexes with response elements on DNA. Estimates of regional induction were converted to color intensities to "paint" the two-dimensional liver for a visual comparison with immunohistochemical observations. To obtain sharp moving boundaries of induced areas with increasing dose (as noted in various experiments), n values in each subcompartment must be large. To create realistic total induction curves that are relatively smooth, the differences in Kd values between adjacent subcompartments must be less than fivefold. Because of the high n values, the low-dose induction characteristics predicted with the multicompartment liver model differ significantly from those predicted with a model that considers the liver as a single homogeneous compartment.

Animals↗
PubMed1994

Heinrich Wieland--prize lecture. Transport of proteins across mitochondrial membranes.

The vast majority of proteins comprising the mitochondrion are encoded by nuclear genes, synthesized on ribosomes in the cytosol, and translocated into the various mitochondrial subcompartments. During this process proteins must cross the lipid membranes of the mitochondrion without interfering with the integrity or functions of the organelle. In recent years an approach combining biochemical, molecular, genetic, and morphological methodology has provided insights into various aspects of this complex process of intracellular protein sorting. In particular, a greater understanding of the molecular specificity and mechanism of targeting of mitochondrial preproteins has been reached, as a protein complex of the outer membrane which facilitates recognition and initial membrane insertion has been identified and characterized. Furthermore, pathways and components involved in the translocation of pre-proteins across the two mitochondrial membranes are being dissected and defined. The energetics of translocation and the processes of unfolding and folding of proteins during transmembrane transfer are closely linked to the function of a host of proteins known as heat-shock proteins or molecular chaperones, present both outside and inside the mitochondrion. In addition, the analysis of the process of folding of polypeptides in the mitochondrial matrix has allowed novel and unexpected insights into general pathways of protein folding assisted by folding factors. Pathways of sorting of proteins to the four different mitochondrial subcompartments--the outer membrane (OM), intermembrane space, inner membrane (IM) and matrix--are only partly understood and reveal an amazing complexity and variation. Many additional protein factors are involved in these latter processes, a few of which have been analyzed, such as cytochrome c heme lyase and cytochrome c1 heme lyase, enzymes that catalyze the covalent addition of the heme group to cytochrome c and c1 preproteins, and the mitochondrial processing peptidase which cleaves signal sequences after import of preproteins into the matrix. Thus, the study of transport of polypeptides through the mitochondrial membranes does not only contribute to the understanding of how biological membranes facilitate the penetration of macromolecules but also provides novel insights into the structure and function of this organelle.

Animals↗
PubMed1991

Microassays for androgen and progesterone receptor quantitation as compared with standard saturation analyses in human prostatic tissues.

Simultaneous measurement of androgen and progesterone receptor content in cytosol and salt extractable nuclear subcompartments of benign hyperplastic prostatic tissue was carried out with various microassay techniques and compared to the results from analyses on bulky tissue from the same tissue specimens. The microassays were carried out as modified saturation analyses or as single concentration assays at various degrees of dilution with tris-EDTA-glycerol (TEG) buffer. Tissue samples for the standard assay weighed between 1.76 and 3.22 g, whereas the microassay samples weighted between 0.14 and 0.47 g. When considering the results of the standard assay as the "true" value, the microassays on the same tissue samples tended to underestimate both the androgen and progesterone receptor contents. Data from the microassays showed a wide variation of the androgen and progesterone receptor content in cytosol and nuclei. With the standard assay technique no detectable amount of progesterone receptor was found in the nuclei, whereas the microassays often indicated false-positive progesterone receptor content in this subcompartment. Therefore, the measurements of steroid receptors using biochemical microassays in prostatic tissue are unreliable and not suitable for clinical use, at least with the techniques available today. Reports in the literature based on such assays should therefore be interpreted with great caution.

Cytosol↗
PubMed1986

Ultrastructural localization of the major proteoglycan and type II procollagen in organelles and extracellular matrix of cultured chondroblasts.

The mechanisms of synthesis and intracellular routing of the various cartilage matrix macromolecules are still unclear. We have studied this problem in cultured chondroblasts at the ultrastructural level using monospecific antibodies against the core protein of the keratan sulfate/chondroitin sulfate-rich cartilage proteoglycan (KS:CS-PG) or Type II procollagen, and cuprolinic blue, a cationic dye that binds to the glycosaminoglycan chains of proteoglycans. Intracellularly, the proteoglycan antibodies localized KS:CS-PG and its precursors primarily in the Golgi complex and secretory vesicles. In contrast, the bulk of Type II procollagen was found within the rough endoplasmic reticulum (ER). While devoid of collagen, the extracellular matrix was rich in KS:CS-PG molecules some of which studded the chondroblast plasmalemma. Cuprolinic blue staining indicated that the proteoglycans present in the Golgi complex fell into a predominant class of large proteoglycans, probably representing KS:CS-PG, and a minor class of smaller proteoglycans. Groups of these divergent proteoglycans often occupied distinct Golgi subcompartments; moreover, single large proteoglycans appeared to align along the luminal surface of Golgi cisternae and secretory vesicles. These results suggest that in cultured chondroblasts KS:CS-PG and Type II procollagen are differentially distributed both in organelles and in the extracellular matrix, and that different proteoglycan types may occupy distinct subcompartments in trans Golgi.

Animals↗
PubMed1995

Morphological plasticity of axotomized retinal ganglion cells following intravitreal transplantation of a peripheral nerve segment.

During normal development of retinal ganglion cells when the axons are growing, transient dendritic spines have been observed. Similar dendritic spine-like processes are also exhibited by retinal ganglion cells undergoing axonal regeneration into a peripheral nerve grafted to the damaged optic axons. Here we show, using the intracellular injection of Lucifer Yellow, that when a segment of peripheral nerve is transplanted to the vitreous body, a procedure which induces ectopic sprouting of axon-like processes from the cell bodies and dendrites of some retinal ganglion cells, similar spine-like processes appear on the dendrites of cells with ectopic sprouts. Quantitative analysis indicated that there were significant changes with posttransplantation survival time in the distributions of spine-like processes and axon-like processes on these sprouting retinal ganglion cells following the intravitreal transplantation of a piece of peripheral nerve. The remodelling of the spine-like processes and axon-like processes correlated with one another suggesting that plastic changes can occur in certain dendritic subcompartments independent of the growth activity of the other dendritic subcompartments.

Animals↗
PubMed1996

Actin and the coordination of protrusion, attachment and retraction in cell crawling.

To crawl over a substrate a cell must first protrude in front, establish new attachments to the substrate and then retract its rear. Protrusion and retraction utilise different subcompartments of the actin cytoskeleton and operate by different mechanisms, one involving actin polymerization and the other myosin-based contraction. Using as examples the rapidly locomoting keratocyte and the slowly moving fibroblast we illustrate how over expression of one or the other actin subcompartments leads to the observed differences in motility. We also propose, that despite these differences there is a common coordination mechanism underlying the genesis of the actin cytoskeleton that involves the nucleation of actin filaments at the protruding cell front, in the lamellipodium, and the relocation of these filaments, via polymerization and flow, to the more posterior actin filament compartments.

Actins↗
PubMed1996

Proliferative heterogeneity of human renal cell carcinomas and prevalence of ras gene point mutations.

The variable prevalence and a possible stage-dependent increase of ras gene point mutations in human tumors might correspond to clonal growth advantages of ras-activated cells. Tumor areas with activated ras genes might thus differ in proliferative activity from those lacking ras gene activation. This hypothesis is studied in a series of human renal cell carcinomas that had been used previously for an analysis of proliferative compartments after post-operative vascular [3H]/[14C]thymidine perfusion [Rabes et al. (1979) Cancer 44: 799-813]. The growth fraction of different subcompartments of these tumors was studied by immunohistochemistry with mib1 antibody, recognizing a fixation- and embedding-resistant epitope of Ki-67 protein. Thirty subpopulations of 14 human renal cell carcinomas that exhibited a broad spectrum of proliferative activity were chosen for an analysis of the prevalence of K-ras point mutations in exon 1 by a mutation-enriching primer-mediated restriction-fragment-length-polymorphism analysis and/or direct sequencing of polymerase-chain-reaction-amplified material. The combined autoradiographic and immunohistochemical analysis confirmed the intra- and intertumoral proliferative heterogeneity. Compared to [3H]/[14C]thymidine labeling indices, mib1 labeling indices are higher. The ratio of mib1 to [3H]/[14C]thymidine labeling indices varies from 1.9 to 4.1 for the individual tumor subcompartments. However, neither in K-ras codons 12/13 nor in adjacent codons did we detect any mutations in the various tumor compartments. The results suggest that neither mode of proliferation nor type of differentiation is related to K-ras exon 1 point mutations in human renal cell carcinomas.

Adult↗
PubMedDate not supplied

Endocytic routes to the Golgi apparatus.

The endocytic routes of labelled lectins as well as cationic ferritin were studied in cells with a regulated secretion, i.e. pancreatic beta cells, and in constitutively secreting cells, i.e. fibroblasts and HepG2 hepatoma cells, paying particular attention to routes into the Golgi apparatus. Considerable amounts of internalised molecules were taken up into the trans Golgi network (TGN) and into Golgi subcompartments in all three cell types as well as in secretory granules of the pancreatic beta cells. The internalised materials did not pass rapidly the TGN and Golgi stacks, but were still present hours after internalisation, being then particularly concentrated in TGN-elements and in the transmost Golgi cisterna. Endocytosed materials reached forming secretory granules present in the TGN. Further, direct fusion between endocytotic vesicles and mature secretory granules was observed. Golgi subcompartments as well as endocytic TGN containing endocytosed materials were in close apposition to specialised regions of the endoplasmic reticulum. The Golgi apparatus including its parts containing endocytosed materials were transformed into a tubular reticulum upon treatment with the fungal metabolite Brefeldin A. Rarely, internalised material was observed in the lumen of the endoplasmic reticulum, thus providing evidence for an endocytic plasma membrane to endoplasmic reticulum route.

Animals↗
PubMed1992

Dynamics of vinblastine-induced autophagocytosis in murine pancreatic acinar cells: influence of cycloheximide post-treatments.

Accumulation of autophagic vacuoles (AVs) was monitored by electron microscopic morphometry in murine pancreatic acinar cells during the 5-hr period after a single injection of vinblastine (VBL). The expansion of the autophagic compartment (AC) occurred in two waves. AVs accumulated in the first 90 min and regressed in the next hour, but thereafter AC expanded again, and 5 hr following the VBL injection, as much as 5.3% of the cytoplasmic volume was found sequestered into the AC. The high rates of accumulation of AVs indicated that VBL stimulated AV formation (segregation) during both expansion phases. To have a deeper insight into the dynamics of the process segregational inhibitor cycloheximide (CHI) was given 1 and 3 hr after VBL and the subsequent regression of the AC and its subcompartments (i.e., early, advanced, and late AVs) were measured during the next 90 min. We found that regression of AVs was fast in the first expansion and slowed down in the second expansion phase during which only early AVs regressed. CHI proved to be a fast and effective inhibitor of autophagic segregation, whether it was given before, simultaneously, or after the VBL injection. The aforementioned results argue for a dual mode of action of VBL (i.e., a prompt stimulation of segregation and a delayed retardation of AV maturation). The two effects of the alkaloid prevail differently along the time course. A further analysis of the behavior of the AC subcompartments showed that CHI perhaps inhibits segregational step(s) occurring prior to the actual formation of the autolysosomes.

Animals↗
PubMed1996

Effects of the structure of a toxicokinetic model of butadiene inhalation exposure on computed production of carcinogenic intermediates.

A flow-limited physiologically based toxicokinetic model was constructed for uptake, metabolism, and clearance of butadiene (BD) and its principal metabolite 1,2-epoxy-3-butene (EB), using physiological and biochemical parameters from the literature where available. The model includes compartments for blood, liver, lung, fat, GI tract, other rapidly perfused tissues, and slowly perfused tissues. The blood was distributed among compartments for arterial plus venous blood and subcompartments for vascular spaces associated with each of the tissue compartments. The lung contained a subcompartment for the alveolar space. Metabolic activation of BD by cytochrome P450-catalyzed epoxidation was modeled as occurring in liver, lung, and the rapidly perfused tissue compartments. The detoxication of EB catalyzed by epoxide hydrolase and glutathione S-transferase (GST) was modeled as occurring in liver, lung, and the rapidly perfused tissues compartments and by blood GST activity. The model also includes depletion of glutathione (GSH) by GST-catalyzed conjugation of EB and 3-butene-1,2-diol and resynthesis of GSH from cysteine. Values of biochemical parameters that were unavailable in the literature were estimated by iteratively reweighted least squares optimization to reproduce data for uptake of BD and EB by rats and mice in closed chambers. The resulting model also reproduced the depletion of GSH in liver and lung in flow-through systems. It reproduced the concentrations of expired EB produced from BD in closed chambers but overpredicted separately measured blood EB concentrations in flow-through systems, indicating an inconsistency between these two experiments that cannot be resolved by this model or an inadequacy in the model. Equilibration of chamber gases with the alveolar space and alveolar gas with lung capillary blood results in much less dilution of the inhaled gas in the blood compared with the predictions of models in which chamber gas equilibrates directly with the total circulation. The production of EB predicted by the present model was found to be sensitive to a number of physiological and biochemical parameters. A valid and useful toxicokinetic model must have reliable physiological and enzymological data for BD biotransformation before it can be credibly used for human risk assessment.

Administration, Inhalation↗
PubMed1982

Protein degradation in rat liver. Evidence for populations of protein degradation rates in cellular organelles.

Protein degradation in liver subcellular and submitochondrial fractions from adult rats has been measured by a double isotope technique. Protein subunits have been resolved by two-dimensional polyacrylamide gel electrophoresis. No consistent relationship between subunit size or isoelectric point and degradation rate has been found in any fraction. Frequency analysis of isotope ratios for protein subunits in subcellular and submitochondrial fractions has been carried out. The analyses demonstrate that degradation rates of resolved protein subunits in organelles or organelle subcompartments can be grouped into populations. Resolved protein subunits in subcellular and submitochondrial fractions can be grouped into one, two or three populations. The results suggest limited heterogeneity of protein degradation rates within each subcellular organelle and support the notion that morphological subcompartment rather than molecular property may be the prime determinant of rates of degradation of individual proteins.

Animals↗