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At least 649 records · Page 36Linked to original sources
PubMedDate not supplied

Simple radioactive assay for the estimation of DNA breaks.

The intactness of DNA is an important part of the normal cellular structure. Any change to the DNA in the form of breaks leads to a change in the integrity, which in turn leads to abnormality in the cellular activity. Many discrepancies have been reported among the various methods of detecting DNA damage. Here, a simple, sensitive and reproducible method has been developed for the detection of DNA breaks by radioactive labelling of 5' broken ends. The method was evaluated by studying chemically induced DNA damage by using both organochloride (2,4-dichlorophenoxyacetic acid and lindane) and organophosphorus (sevin and phosphamidon) compounds at different concentrations. Phosphamidon, one of the organophosphorus compounds studied, showed complete degradation of the DNA after treatment. Radioactive analysis of phosphamidon showed higher counts at the lowest concentration (20 microg) of the chemical when compared with the control (2752 scintillation counts per minute, scm). Studies on the chemically induced DNA breaks by radiolabelling revealed that the cumulative effect of the organophosphorus and organochloride compounds showed maximum counts in all the samples (the highest being 2904 scm) when compared with the organophosphorus and organochloride compounds studied separately (the highest being 1881 and 2260 scm, respectively). Radiolabelling studies on the blood samples of 23 pesticide workers by the newly developed assay showed a significant positive correlation (0.893) between the number of years of exposure and the scintillation counts. A maximum of 11,702 scm (for 18 years of exposure) and a minimum of 1682 scm (for 4 years of exposure) were recorded compared with 1253 scm for the negative control. This method can be used effectively for estimation of the DNA breaks, irrespective of its nature.

Agricultural Workers' Diseases↗
PubMed2006

Force meets chemistry: analysis of mechanochemical conversion in focal adhesions using fluorescence recovery after photobleaching.

Mechanotransduction--the process by which mechanical forces are converted into changes of intracellular biochemistry--is critical for normal cell and tissue function. Integrins facilitate mechanochemical conversion by transferring physical forces from the extracellular matrix, across the cell surface, and to cytoskeletal-associated proteins within focal adhesions. It is likely that force alters biochemistry at these sites by altering molecular binding affinities of a subset of focal adhesion proteins, but this has been difficult to quantify within living cells. Here, we describe how the fluorescence recovery after photobleaching (FRAP) technique can be adapted and used in conjunction with mathematical models to directly measure force-dependent alterations in molecular binding and unbinding rate constants of individual focal adhesion proteins in situ. We review these recent findings, and discuss the strengths and limitations of this approach for analysis of mechanochemical signaling in focal adhesions and other cellular structures. The ability to quantify molecular binding rate constants in the physical context of the living cytoplasm should provide new insight into the molecular basis of cellular mechanotransduction. It also may facilitate future efforts to bridge biological experimentation and mathematical modeling in our quest for a systems biology level description of cell regulation.

Animals↗
PubMed1998

Lysosomal enzyme trafficking between phagosomes, endosomes, and lysosomes in J774 macrophages. Enrichment of cathepsin H in early endosomes.

In this study we take advantage of recently developed methods using J774 macrophages to prepare enriched fractions of early endosomes, late endosomes, dense lysosomes, as well as phagosomes of different ages enclosing 1-micron latex beads to investigate the steady state distribution and trafficking of lysosomal enzyme activity between these organelles. At steady state these cells appear to possess four different cellular structures, in addition to phagolysosomes, where acid hydrolases were concentrated. The first site of hydrolase concentration was the early endosomes, which contained the bulk of the cellular cathepsin H. This enzyme was acquired by phagosomes significantly faster than the other hydrolases tested. The second distinct site of lysosomal enzyme concentration was the late endosomes which contain the bulk of cathepsin S. The third and fourth large pools of hydrolases were found in two functionally distinct types of dense lysosomes, only one of which was found to be secreted in the presence of chloroquine or bafilomycin. Among this secreted pool was soluble furin, generally considered only as a membrane-bound trans-Golgi network resident protein. Thus, the organelles usually referred to as "lysosomes" in fact encompass a growing family of highly dynamic but functionally distinct endocytic organelles.

Animals↗
PubMed2001

The concept of self-organization in cellular architecture.

In vivo microscopy has recently revealed the dynamic nature of many cellular organelles. The dynamic properties of several cellular structures are consistent with a role for self-organization in their formation, maintenance, and function; therefore, self-organization might be a general principle in cellular organization.

Animals↗
PubMed1980

Evaluation of the polyene antibiotic filipin as a cytochemical probe for membrane cholesterol.

Distribution of the filipin-cholesterol complexes has been studied by freeze-fractures techniques. While filipin-cholesterol complexes are formed in fixed as well as unfixed cells, the distribution of the complexes varies with fixation. Furthermore, filipin induces dramatic rearrangement of intramembrane particles in unfixed, but not in fixed, cells. Examination of thin sections of filipin-treated cells shows that filipin alone induces substantial cellular damage, while filipin treatment simultaneous to fixation or following fixation does not cause noticeable damage. For filipin to be an effective cytochemical probe of membrane structure, cellular damage must be minimized; this is best achieved by treating cells with filipin following glutaraldehyde fixation.

Animals↗
PubMed2006

Effects of microgravity on cell cytoskeleton and embryogenesis.

The aim of this review is to compile, summarize and discuss the effects of microgravity on embryos, cell structure and function that have been demonstrated from data obtained during experiments performed in space or in altered gravity induced by clinostats. In cells and tissues cellular structure and genetic expression may be changed in microgravity and this has a variety of effects on embryogenesis which include death of the embryo, failure of neural tube closure, or final deformities to the overall morphology of the newborn or hatchling. Many species and protocols have been used for microgravity space experiments making it difficult to compare results. Experiments on the ways in which embryonic development and cell interactions occur in microgravity could also be performed. Experiments that have been done with cells in microgravity show changes in morphology, cytoskeleton and function. Changes in cytoskeleton have been noted and studies on microtubules in gravity have shown that they are gravity sensitive. Further study of basic chemical reactions that occur in cells should be done to shed some light on the underling processes leading to the changes that are observed in cells and embryos in microgravity.

Animals↗
PubMed1986

S.E.M. study I: Gastric and duodenal lesions induced by non-steroidal anti-inflammatory drugs (aspirin, piroxicam) in man.

A scanning electron microscopy (SEM) study was performed on the gastric and duodenal mucosa of 14 patients treated with Aspirin or Piroxicam. Six patients with normal mucosal morphology were treated with either 1.5 g/day of Aspirin (3 patients) or 20 mg/day or Piroxicam (3 patients) for one month. In addition, 8 rheumatic patients were treated with a similar dose of Aspirin (4 patients) or Piroxicam (4 patients) for at least 4 months. SEM was used to evaluate the following parameters: mucosal structure, cellular exfoliation and anisocytosis, alteration of the microvilli (blebs). The mucosa showed various aspects of alterations ranging from minimal changes (microvillar alteration) to a completely subverted structure both in the stomach and in the duodenum. Mucosal changes, both on the edge of the macroscopic lesions and at a distance from these, were visible with SEM even when endoscopy was normal. Piroxicam appeared slightly less damaging than Aspirin, and no difference was observed between short-term and long-term treated patients, either with Aspirin or with Piroxicam.

Adult↗
PubMedDate not supplied

The natural subcellular surface structure of the bovine sperm cell.

We used atomic force microscopy (AFM), which utilizes a novel 3D image-contrast mechanism, to obtain nanometer-resolved, topographic data images of the natural surface structures of untreated bovine sperm cells. Freshly ejaculated, thawed, sonicated, and demembranated bovine sperm were adsorbed passively or by motility from suspension onto a coverglass substrate and directly imaged in normal air and saline environments without damaging the cells. Our AFM images of the surface structures of unfixed sperm imaged in normal air were consistent with previous electron microscope results on frozen or fixed sperm, demonstrating that the accurate preservation of small cellular structures is achievable using greatly simplified AFM sample preparation and imaging environments. Our AFM results also indicate that imaging sperm in physiologic buffer provides more native views of sperm due to the retention of cytoplasmic structures easily disrupted by drying forces. In addition, the AFM images show that numerous nanometersized subcellular structures of the sperm head and tail regions could be clearly visualized on rapidly prepared, unfixed, intact cells. Consequently, AFM should be considered a new tool for studying sperm structure abnormalities and monitoring the specific effects of, or damage caused by, various chemical reactants or other treatments on the structures of metabolically active or partially demembranated sperm. AFM is now emerging as an important new structural technique for imaging hydrated cells and organelles and, in addition, has the capabilities to physically "interrogate" them with the local probe.

Acrosome↗
PubMed2026

Single-cell and spatial transcriptomic technologies for lung cancer tumor microenvironment analysis.

Lung cancer remains one of the leading causes of cancer-related mortality worldwide; beyond its rising incidence, its marked molecular heterogeneity and complex tumor microenvironment (TME) hinder treatment response and drive resistance, contributing directly to its high mortality rate. Single-cell RNA sequencing (scRNA-seq) and spatial transcriptomics (ST) provide complementary approaches for dissecting these features. scRNA-seq enables high-resolution analysis of cellular diversity and transcriptional states but requires tissue dissociation and therefore loses spatial context. In contrast, ST preserves tissue architecture and provides insights into how gene-expression programs within the TME are organized, although no currently available spatial platform combines whole-transcriptome coverage with true single-cell resolution over large tissue areas. Together, these technologies have enabled detailed mapping of tumor, immune and stromal populations and of their spatial interactions, revealing functionally distinct cellular niches that contribute to immune evasion, metastasis and response to therapy. In this narrative review we organize the primary literature around a single question, how spatially structured cellular ecosystems, rather than individual cell types, determine therapeutic response and resistance in lung cancer - and we explicitly separate observations that are reproducible across independent cohorts and platforms from those that remain confined to single studies. We further summarize the technical, analytical and logistic barriers that currently prevent spatially resolved signatures from entering routine diagnostic pathology. Understanding dysregulated pathways and spatially constrained intercellular communication within the TME helps identify candidate biomarkers and may support the identification of therapeutic approaches directed at tumor-intrinsic programs as well as at microenvironment-driven resistance mechanisms.

Cell-cell communication↗
PubMed1998

Aging and the liver: an update.

The issue of whether or not liver function is compromised in the elderly population remains unresolved. Numerous age-related changes in hepatic structure and function have been described, but many of these observations are qualitative, were made under suboptimal experimental conditions, or are simply contradictory. Changes in hepato-cellular structural parameters, e.g., increased hepatocyte size, increase in the number of binucleated cells, altered mitochondria, and endoplasmic reticulum, have been reported. However, quantitative morphological analyses have refuted many of these observations. There are few functional data that correlate with structural changes. Serum and biliary cholesterol appear to rise, predisposing elderly people to increased incidences of coronary disease and gallstones, respectively. The rate of liver regeneration declines in old animals, but the regenerative capacity remains unchanged, perhaps reflecting an age-associated reduction in the response to hepatotrophic factors. This senescent change has important clinical implications with regard to surgical intervention for liver disease, e.g., resection or transplantation. Nevertheless, most outcomes studies suggest that age alone should not be a determining factor in such clinical decisions. Geriatric patients exhibit a decline in the hepatic clearance of certain drugs and a marked increase in the frequency of adverse drug reactions, reflecting an increase in polypharmacy regimens and declines in liver volume and blood flow rather than reduced Phase I metabolism. Although the livers of elderly subjects are characterized by a decline in adaptive responsiveness and reduced reserve capacity, clinical tests suggest that liver function is well-maintained in this age group.

Aging↗
PubMed1999

Osmosensing by bacteria: signals and membrane-based sensors.

Bacteria can survive dramatic osmotic shifts. Osmoregulatory responses mitigate the passive adjustments in cell structure and the growth inhibition that may ensue. The levels of certain cytoplasmic solutes rise and fall in response to increases and decreases, respectively, in extracellular osmolality. Certain organic compounds are favored over ions as osmoregulatory solutes, although K+ fluxes are intrinsic to the osmoregulatory response for at least some organisms. Osmosensors must undergo transitions between "off" and "on" conformations in response to changes in extracellular water activity (direct osmosensing) or resulting changes in cell structure (indirect osmosensing). Those located in the cytoplasmic membranes and nucleoids of bacteria are positioned for indirect osmosensing. Cytoplasmic membrane-based osmosensors may detect changes in the periplasmic and/or cytoplasmic solvent by experiencing changes in preferential interactions with particular solvent constituents, cosolvent-induced hydration changes, and/or macromolecular crowding. Alternatively, the membrane may act as an antenna and osmosensors may detect changes in membrane structure. Cosolvents may modulate intrinsic biomembrane strain and/or topologically closed membrane systems may experience changes in mechanical strain in response to imposed osmotic shifts. The osmosensory mechanisms controlling membrane-based K+ transporters, transcriptional regulators, osmoprotectant transporters, and mechanosensitive channels intrinsic to the cytoplasmic membrane of Escherichia coli are under intensive investigation. The osmoprotectant transporter ProP and channel MscL act as osmosensors after purification and reconstitution in proteoliposomes. Evidence that sensor kinase KdpD receives multiple sensory inputs is consistent with the effects of K+ fluxes on nucleoid structure, cellular energetics, cytoplasmic ionic strength, and ion composition as well as on cytoplasmic osmolality. Thus, osmoregulatory responses accommodate and exploit the effects of individual cosolvents on cell structure and function as well as the collective contribution of cosolvents to intracellular osmolality.

ATP-Binding Cassette Transporters↗
PubMedDate not supplied

Normal ultrastructure of the kidney and lower urinary tract.

The mammalian urinary tract includes the kidneys, ureters, urinary bladder, and urethra. The renal parenchyma is composed of the glomeruli and a heterogeneous array of tubule segments that are specialized in both function and structure and are arranged in a specific spatial distribution. The ultrastructure of the glomeruli and renal tubule epithelia have been well characterized and the relationship between the cellular structure and the function of the various components of the kidney have been the subject of intense study by many investigators. The lower urinary tract, the ureters, urinary bladder, and urethra, which are histologically similar throughout, are composed of a mucosal layer lined by transitional epithelium, a tunica muscularis, and a tunica serosa or adventitia. The present manuscript reviews the normal ultrastructural morphology of the kidney and the lower urinary tract. The normal ultrastructure is illustrated using transmission electron microscopy of normal rat kidney and urinary bladder preserved by in vivo perfusion with glutaraldehyde fixative and processed in epoxy resin.

Animals↗
PubMed1991

[Cytologic diagnosis of thyroid gland adenoma and goiter].

Examinations of smear impressions of postoperative biopsy specimens of the thyroid from adenoma patients have revealed among polygonal structures 1 to 20 spherical complexes (per micropreparation) consisting of relatively monomorphic follicular cells with intensively blue cytoplasm. The 'ball' size was 200-900 micron or more. Cellular composition, specific features of cellular structure, and the presence of spherical structures permit differentiating adenoma from goiter.

Adenoma↗
PubMedDate not supplied

[Antioxidants: the therapy of the future?].

In the present review piece, we analyze the formation of free radicals as a consequence of the cellular metabolism in aerobe organism, and the beneficious and harmful actions thereof on cellular structures. The balance existing between free radicals and the so-called antioxidant defenses, is a key factor for preventing the development of noxious processes at the cellular and tissue level. In accordance with the present scientific knowledge, the excessive production of free radicals in the organism, and the imbalance between the concentrations of these and the antioxidant defenses, may be related to processes such as aging and several diseases, among which we find cancer, ischemic processes, senile dementia, diabetes, pulmonary and pancreatic diseases, lupus erythematosus, cirrhosis, intestinal inflammatory disease, multiple sclerosis, arthritis, arteriosclerosis, cardiovascular disease, diseases of the central nervous system and the brain. According to the results of numerous research works conducted with the administration of several molecules with an antioxidant activity, one is beginning to see what their role will be in the pharmacological therapeutics for the treatment of a large number of patients such as those with burns, traumas, septics, shock, surgery, transplantation, radiation or chemotherapy, respiratory distress syndrome, AIDS, etc. We may possibly be facing a therapeutic tool which is of great interest in the clinical area, which shall be developed in the near future, as clinical trials which permit confirmation of their efficacy are conducted.

Animals↗
PubMed2004

Collagenase predigestion on paraffin sections enhances collagen immunohistochemical detection without distorting tissue morphology.

Reliable immunohistochemical detection of collagen in formalin fixed, paraffin embedded tissues requires protease digestion. While these pan-proteases (pepsin, trypsin, protease K, etc.) enhance collagen detection, they also digest many other tissue proteins and produce poor cellular morphology and unrecognizable cellular structures. Balancing the conditions (protease type, concentration, incubation time and temperature) to digest some, but not all, proteins in a tissue section while optimizing collagen detection requires one to compromise improved collagen immunolabeling with adequate cellular morphology. Furthermore, optimal conditions for digesting tissue proteins to enhance collagen detection vary among tissue types and their fixation. Although brain is not typically subject to these deleterious consequences, structures such as epithelium, spermatids, stroma etc. and other tissues with complicated histology are profoundly affected. To resolve this technical dilemma, we discovered a novel use for collagenase to enhance collagen immunodetection without affecting the noncollagen proteins, thereby preserving tissue morphology. Collagenase, which is typically used in vitro for disassociation of cells, has never been used reliably on formalin fixed, paraffin embedded tissue sections. This new use of collagenase for immunohistochemistry promotes increased collagen immunolabeling, is easy to use, is versatile, and allows preservation of tissue structure that provides maximal and accurate histological information.

Cell Size↗
PubMed2001

Structural, biochemical and signaling properties of the low-density lipoprotein receptor gene family.

The low-density lipoprotein (LDL) receptor (LDL-R) family members (LDL-R, LRP, megalin, VLDL-R, apoER2) bind several extra-cellular structurally dissimilar ligands and internalize them for degradation by lysosomes by a process called receptor-mediated endocytosis. The receptor-mediated endocytosis involves immobilization of circulating ligands onto the cell-surface followed by their internalization and degradation. All the receptors can perform both of these functions. However, in the majority of the cases, other proteins immobilize ligands on to the cell-surface and subsequent internalization is mediated by these receptors. The LDL-R and LRP play important roles in plasma cholesterol homeostasis and fetal development. Megalin is an antigenic determinant for Heymann nephritis in rats and may be important for re-absorption of various molecules by the kidney. VLDL-R homologue in chicken is essential for female fertility. This receptor and apoER2 are critical for the proper development of the brain in mice. The members of the LDL-R gene family contain several complement-type and EGF precursor-like repeats, and single transmembrane and cytoplasmic domain. Cysteine-rich complement-type repeats containing DxSDE sequences at the C-termini constitute ligand-binding domains. In contrast to the ligand binding domains, receptor-binding domains in different ligands do not share sequence homology. It has been proposed that positive electrostatic surface potentials, not the primary sequences, in different ligands constitute receptor-binding domains. The EGF precursor homology repeats in receptors are important for the dissociation of ligands from receptors in endocytic vesicles. The transmembrane domain is necessary for anchoring to membranes and the cytoplasmic domain is required for their targeting to coated pits and subsequent internalization. The receptor-mediated endocytosis involves recognition of the NPXY motif by clathrin. Recently, this motif has also been implicated in signaling pathways that are crucial in brain development. The signaling process involves the recognition of the NPXY motif by Disabled-1 protein and possibly other proteins involved in intracellular signaling cascade. The LDL-R gene family has provided important insights into the mechanisms of ligand catabolism and may serve as new targets for the treatment of different cardiovascular and neuronal disorders. In the future, their role in signaling may provide novel insights into brain development and neuronal layering.

Amino Acid Motifs↗
PubMed1977

Physiology of aging: metabolic changes during the climacteric and menopausal periods.

Endocrine regulation is altered with age. Changes in hormone secretory rates, patterns of secretion, and responses to physiologic and pharmacologic stimuli have been demonstrated. Target tissue sensitivity to hormones appears to be age related and to involve hormone receptor mechanisms, enzymatic responses, and carrier proteins. The aging process reflects changes in cellular structure and metabolism that decrease the function and efficiency of tissues. Structural changes occur with age and are characterized by a loss of elastic tissue, accumulation of highly crosslinked collagen, reduction in ground substance, and an increase in a cellular fluorescent pigment called lipofuscin. These changes decrease the pliability of tissues and alter membrane permeability. Modifications in protein synthesis and enzymatic systems may predispose the aged to autoimmune and neoplastic disease. Endocrine diseases are common in the aged and often may be atypical in their presentation. Cancer is common in the elderly and may be associated with endocrine manifestations because the tumor is producing hormones. Physicians must be aware of these syndromes so that a correct diagnosis can be established, treatment instituted, and the quality of life improved. Laboratory test results in the aged must be carefully interpreted because most aged patients are taking medications that interfere with endocrine tests and also the normal range for the elderly has not been clearly established. Abnormal laboratory diagnostic tests are common in the aged and do not always imply endocrine disease. No hormone can reverse the natural process of aging; however, hormones do regulate all metabolic functions and the neuroendocrine system plays a key role in all phases of growth, maturation, and aging.

Adrenal Gland Diseases↗
PubMed2005

Inhibition of protein kinases C prevents murine cytomegalovirus replication.

For successful establishment of infection and initiation of the replication cycle, murine cytomegalovirus (MCMV) utilizes cellular structures and functions, including cell-membrane penetration, capsid dismantling and cytosolic transport of viral DNA into the nucleus. These early events of MCMV infections are dependent on cellular regulatory mechanisms, primarily protein phosphorylation. In the present study, protein kinase inhibitors were used to explore the role of protein phosphorylation mediated by protein kinases C (PKCs) in the very early events of MCMV infection. Inhibitory effects were determined by immunofluorescence and Western blot analysis of MCMV IE1 and E1 protein expression and by production of infectious virions in cell culture. It was found that H-7, a broadly specific inhibitor of cellular protein kinases, prevented virus replication in a dose-dependent and reversible manner, and that the block in replication occurred very early in infection. More specific PKC inhibitors (sangivamycin, calphostin C and bisindolylmaleimide II), Ca(2+)/calmodulin inhibitors (EDTA and W7) and phorbol esters (PMA) were used to dissect PKC-subclass contribution in the very early events of MCMV replication. The results indicate that the role of diacylglycerol/phorbol ester-dependent but calcium-independent PKCs is essential for establishment of MCMV infection in the host cell, starting at a very early stage of infection.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗