In situ identification of cellular drug targets in mammalian tissue

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Localization and activity of iNOS in normal human lung tissue and lung cancer tissue

Inducible nitric oxide synthase (iNOS) is one of three enzymes generating nitric oxide (NO) from the amino acid L-arginine. iNOS-derived NO plays an important role in several physiological and pathophysiological conditions. NO is a free radical which produces many reactive intermediates that account for its bioactivity. In the human lung, the alveolar macrophage is an important producer of cytokines and this production may be modified by NO. Moreover, high concentrations of NO have been shown to increase nuclear factor kappaB (NF-kB) activation. Recent investigations of NO expression in tumor tissue indicated that, at least for certain tumors, NO may mediate one or more roles during the growth of human cancer. We have studied iNOS in two tissue groups: normal human lung tissue and human lung cancer tissue. We localized iNOS in these tissues by immunohistochemistry and tested the mRNA expression by RT-PCR, the protein level by Western blot, and the protein activity by radiometric analysis. The results demonstrate different expression, localization and activity of iNOS in normal versus tumor tissue. This is suggestive of a role for NO production from iNOS in human lung cancer because high concentrations of this short molecule may transform to highly reactive compounds such as peroxynitrite (ONOO-); moreover, through the upregulator NF-kB, they can induce a chronic inflammatory state representing an elevated risk for cell transformation to cancer.     

Future prospects for tissue engineered lung transplantation: Decellularization and recellularization-based whole lung regeneration

The shortage of donor lungs for transplantation causes a significant number of patient deaths. The availability of laboratory engineered, functional organs would be a major advance in meeting the demand for organs for transplantation. The accumulation of information on biological scaffolds and an increased understanding of stem/progenitor cell behavior has led to the idea of generating transplantable organs by decellularizing an organ and recellularizing using appropriate cells. Recellularized solid organs can perform organ-specific functions for short periods of time, which indicates the potential for the clinical use of engineered solid organs in the future. The present review provides an overview of progress and recent knowledge about decellularization and recellularization-based approaches for generating tissue engineered lungs. Methods to improve decellularization, maturation of recellularized lung, candidate species for transplantation and future prospects of lung bioengineering are also discussed.     

Cell division and tissue repair following localized damage to the mammalian lung.

Small local wounds on the surface of the mouse lung, produced by cauterization, healed by a typical reparative process involving cell migration and increased cell division in alveolar and bronchial tissues. The local cell division response closely resembled the compensatory cell division response in the same organ which follows unilateral pneumonectomy or unilateral collapse of the lung: initially there was an increase in the rate of DNA synthesis followed by an increased rate of entry into mitosis, both of these functions returning to normal levels within a few days. It is therefore suggested that both types of response are governed by a single regulatory mechanism. The results do not support the view that the rate of cell division is regulated by systemically-circulating mitotic control factors and it is proposed that changes in the cell division rate, both in the reparative and in the compensatory types of response, are determined by local alterations in the concentration of regulatory metabolites. The magnitude of the cell division response was much greater in bronchial than in alveolar tissue, a result which is consistent with the view that new alveolar tissue may be produced by the proliferation and diffentiation of bronchial cells.     

Soft tissue and cellular preservation in vertebrate skeletal elements from the Cretaceous to the present

Soft tissues and cell-like microstructures derived from skeletal elements of a well-preserved Tyrannosaurus rex (MOR 1125) were represented by four components in fragments of demineralized cortical and/or medullary bone: flexible and fibrous bone matrix; transparent, hollow and pliable blood vessels; intravascular material, including in some cases, structures morphologically reminiscent of vertebrate red blood cells; and osteocytes with intracellular contents and flexible filipodia. The present study attempts to trace the occurrence of these four components in bone from specimens spanning multiple geological time periods and varied depositional environments. At least three of the four components persist in some skeletal elements of specimens dating to the Campanian. Fibrous bone matrix is more altered over time in morphology and less likely to persist than vessels and/or osteocytes. Vessels vary greatly in preservation, even within the same specimen, with some regions retaining pliability and other regions almost crystalline. Osteocytes also vary, with some retaining long filipodia and transparency, while others present with short and stubby filipodia and deeply pigmented nuclei, or are pigmented throughout with no nucleus visible. Alternative hypotheses are considered to explain the origin/source of observed materials. Finally, a two-part mechanism, involving first cross-linking of molecular components and subsequent mineralization, is proposed to explain the surprising presence of still-soft elements in fossil bone. These results suggest that present models of fossilization processes may be incomplete and that soft tissue elements may be more commonly preserved, even in older specimens, than previously thought. Additionally, in many cases, osteocytes with defined nuclei are preserved, and may represent an important source for informative molecular data.     

Determination of thirty-two elements in human autopsy tissue

Inductively coupled plasma-mass spectrometry (ICP-MS) was used to determine the concentrations of 32 elements in the human liver and kidney and 20 elements in the bone, obtained from 70 autopsied dead individuals (54 males, 16 females) between 18 and 76 yr of age from the North Bohemia territory of the Czech Republic. Geometric means, median, minimal-maximal range, as well as distribution and correlation analysis were calculated. Some significant differences among tissue concentrations of trace elements of the women and men were found. In the liver, medians of the concentrations of some elements were higher for men than that for women (Al: 770 vs 610 microg/kg; As: 42 vs 27 microg/kg; Cd: 1800 vs 1390 microg/kg; Rb: 3955 vs 3210 microg/kg; V: 160 vs 105 microg/kg). On the contrary, the content of other elements for men was lower (Bi: 0.8 vs 3.2 microg/kg; Cr: 57 vs 72 microg/kg; Hg: 228 vs 325 microg/kg; Zn: 57.1 vs 68.5 mg/kg). In the kidney of men, there were higher contents of Al (360 vs 245 microg/kg) and Hg (135 vs 75 microg/kg) and lower contents of Zn (47.7 vs 59.7 mg/kg) and I (135 vs 220 microg/kg) than those of women. In the case of bone, the concentrations of Cu and Rb were higher for men (1410 microg Cu/kg and 405 microg Rb/kg, respectively) than for women (655 microg Cu/kg and 285 microg Rb/kg, respectively). On the contrary, the content of Mn was considerably lower for men (110 microg Mn/kg) than for women (215 microg Mn/kg).     

Determination of trace elements in human lung samples

Biological Trace Element Research - Lung samples from smokers, nonsmokers, and one stillborn were analyzed by an instrumental neutron activation analysis (INAA) method. Pulmonary tissue and hilum...     

Measurement of the mechanical properties of the human lung tissue

The mechanical properties of human lung tissue were measured in a state of biaxial tension. The experimental data were fitted with a pseudo-elastic constitutive equation proposed earlier and the physical constants were identified.     

Effect of levofloxacine on cell elements of the lung tissue and on the growth of drug resistant Mycobacteria tuberculosis]

Levofloxacin effect on morphokinetic parameters of the murine lung tissue culture was investigated. This model allowed also to evaluate the levofloxacin activity against drug-resistant intracellular and extracellular Mycobacterium tuberculosis: MBC was 0.5-1.0 mg/l. Bactericidal effect started on the 3d day of incubation and was maximal on 7th day. Tissue cells vitality was not changed. Combination of levofloxacin with antimycobacterial drugs of first choice was investigated: isoniazid demonstrated synergistic effect, pyrazinamide also demonstrated synergistic effect. Combination of levofloxacin with rifampicin was antagonistic.     

The analysis of cellular elements in coelomic fluid during early regeneration of of the starfish Asterias rubens L

Three main cell types were found in the coelomic fluid (CF) of intact starfishes: agranulocytes (55-80%) varying in size and form (spherical and ovoid) and with occasional pseudopodia, granulocytes (15-45%), and small cells (up to 2 %) with a high nuclear-cytoplasmic ratio. The starfish response to injury depends on the degree of coelomic fluid loss. After a slight wounding, when only insignificant portion of CF is lost, the cellular composition of circulating fluid changed only slightly. Unlike, a significant injury resulted in rising the share of small cells, regarded presumably as young cells. Besides, after injury the functional characteristics of SF also changed: the proportion of cells with decondensed chromatin and stained nucleoli increased, and coelomocytes acquired ability to form nets at adhesion. Moreover, some new cell types can be found (fusiform cells), with granulocyte proportion in nets increasing. We suppose that after slight wounding circulating coelomocytes may restore from the existing store of differentiated cells beyond the circulation, whereas after significant injury young undifferentiated coelomocytes are involved in the process of restoration.