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A number of different designs of cells have been developed for the study of dissolution processes in packed apatite powders. Basic design requirements were that no deleterious processes, such as high temperature sintering or binding agents, were involved, and that there was maximum opportunity for experimental study. One design used alternating filter paper discs and apatite layers (typically 42 mg). At the end of an experiment, the cell could be disassembled and infrared spectra and X-ray powder diffraction patterns made of individual layers. Cells without filter paper discs were also made, but terminal sampling at well-defined depths was more difficult or impossible. The cells were constructed from poly(methylmethacrylate) so the course of dissolution could be monitored by radiography. Subsurface loss of apatite was almost always seen after from 2 weeks to 8 months exposure to buffer solutions at pH 3.0-5.5. The greatest loss of apatite was typically 0.4-2 mm below the surface, which is at a larger distance than usually seen for dental enamel. This may be attributable to the low packing density (typically 35-60 vol%) found in the present systems compared with enamel. [ABSTRACT FROM AUTHOR]

GPR61 is an orphan receptor that belongs to Class A of G-protein-coupled receptors. It has been reported that GPR61 has a constitutive activity and couples to Gαs. In the present study, we characterized GPR61 function and ligand binding by experimental and molecular docking studies. We demonstrated that heterologous expression of GPR61 in HEK293 cells enhanced the cAMP synthesis response to forskolin, whereas the basal cAMP synthesis was unaffected. 5-(Nonyloxy)tryptamine inhibited forskolin-stimulated cAMP production in GPR61-expressing HEK293 cells. These studies highlight that the intrinsic activity of this receptor is only measurable following its synergy with Gαs. [ABSTRACT FROM AUTHOR]

For the diagnosis of allergy, cellular basophil activation tests (BAT), e.g. histamine or sulfidoleukotriene release tests, have long been introduced, but the expression of basophil activation markers such as CD63 and CD203c detected by flow cytometry has attracted more recent attention. A recent opinion paper in this Journal has stressed not only the potential but also the possible pitfalls of flow-cytometric BAT. We have applied clinical validation of various BAT in various ways for several years, and our experience shows that these new technologies have more potentials and perspectives than pitfalls. A comprehensive review of clinically validated studies on allergy to aeroallergens, insect venoms, latex, food allergens and drugs, e.g. myorelaxants, β-lactams, pyrazolones and non-steroidal anti-inflammatory drugs, as well as chronic urticaria shows clearly that even with different protocols, reproducible and meaningful results can be obtained. Although the available technologies may still be optimized and better standardized, there are no serious reasons to deprive allergic patients of clinically indicated BAT, which can be performed reliably by any laboratory with allergy and flow-cytometric capacity and expertise. Copyright © 2008 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Background/Aims: In thyroid cells, the intracellular pH plays a key role in the control of the iodide uptake, since iodide accumulation is associated with an intracellular acidification. In the present paper we studied the kinetic proprieties of the Na+/H+ antiporter (NHE) and the molecular expression of different NHE isoforms in rat thyroid PC-Cl3 cells. In addition the intracellular buffer capacity was also evaluated. Methods: pHi was measured using the pH sensitive fluorescent dye BCECF-AM. Amiloride, 5-(N,NDimethyl) hydrochloride was used to inhibit NHE activity. RT-PCR and western blot analyses were used to study the expression of NHE mRNA and protein isoforms. Results: PC-Cl3 cells shown a resting pHi, in the absence of CO2/HCO3 -, of 6.94 ± 0.1; after an acid load PC-Cl3 cells recovered toward resting pHi value, using a Na-dependent H+ extrusion mechanisms which was amiloride sensitive (Ki = 23 μM). The kinetic parameters were K(Na)app = 10 ± 2 mM and Vmax = 0.23 ± 0.02 ΔpH/min x 105 cells. NHE1, NHE2 and NHE3 were expressed at the mRNA level as well as at the protein level. Conclusion: PC-Cl3 cells express a functional Na/H exchange activity and different isoforms (NHE1, NHE2 and NHE3) are expressed in the plasma membrane. [ABSTRACT FROM AUTHOR]

The influence of Robertsonian (Rb) heterozygosity on fertility has been the subject of much study in the house mouse. However, these studies have been largely directed at single simple heterozygotes (heterozygous for a single Rb metacentric) or complex heterozygotes (heterozygous for several to many metacentrics which share common chromosome arms). In this paper we describe studies on male multiple simple heterozygotes, specifically the F[sub 1] products of crosses between wild-stock mice homozygous for four or seven metacentrics and wild-stock mice with a standard all-acrocentric karyotype; these F[sub 1] products were characterized by four and seven trivalents at meiosis I, respectively. Mice with the same karyotype, but two different genetic backgrounds were examined. Although a range of meiotic and fertility studies were conducted, particular emphasis was paid to analysis of chromosome pairing, previously not well-described in multiple simple heterozygous mice. The progression of spermatocytes through prophase I was followed by electron microscopy of surface spread material. As previously shown for single simple Rb heterozygotes, the trivalents that characterize multiple simple heterozygotes initially showed delayed pairing of the centromeric region and later showed side arm formation, resulting from non-homologous pairing by the centromeric ends of the acrocentric chromosomes. In the four trivalent groups of mice, 15 and 32% of trivalents showed unpairing in the centromeric region at mid pachytene; equivalent values were 29 and 39% for the seven trivalent groups. Pairing abnormalities (largely attachments and interlocks between trivalents and between a trivalent and the XY configuration) were observed in 18 and 23% of mid pachytene cells in the four trivalent groups and 36 and 49% of cells in the seven trivalent groups. The greater level of pachytene irregularity (unpairing and pairing abnormalities) in seven versus four trivalent heterozygotes was mirrored in terms of higher anaphase I nondisjunction frequency and lower germ cell counts. However, while pachytene irregularities appear to contribute to germ cell death, examples of male sterility in our material undoubtedly also involve genic incompatibilities. Copyright © 2002 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Key events leading to terminal differentiation of preadipocytes into adipocytes have been characterized in the recent years. However, master genes that commit progression from multipotent mesenchymal stem cell to the adipoblast stage of development have not yet been identified. The use of embryonic stem (ES) cells as a route to study early events in adipogenesis and to characterize factors involved in the decision of stem cells to follow the adipogenic pathway is described in this paper. The capacity of lif[sup –] /[sup –] and lifr[sup –] /[sup –] ES cells to undergo adipocyte differentiation is reported as an application of mutant ES cells to study gene function during the development of adipose cells.Copyright © 1999 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

In our previous papers, we have shown that human T cells produce a unique eosinophil chemotactic factor (ECF), termed Ecalectin, with a molecular weight of about 30–50 kD during interaction with BALL-1 (a B cell line) extracts, antigen or mitogen. A 1.6-kB cDNA was isolated from a human T cell-derived expression library that encodes Ecalectin. Ecalectin is a 36-kD protein consisting of 323 amino acids based on its deduced amino acid sequence. Ecalectin was found to be a variant of human galectin-9, a member of the growing family of animal lectins exhibiting affinity for β-galactosides. Recombinant Ecalectin, expressed in COS cells, exhibited potent ECF activity in vitro and in vivo in a dose-dependent manner but not chemotactic activity for neutrophils, lymphocytes, or monocytes. The finding that the Ecalectin transcript is present in various lymphatic tissues and that its expression increases substantially in antigen-activated peripheral blood mononuclear cells suggests that Ecalectin is an important T cell-derived regulator of eosinophil recruitment in allergic reaction sites. [ABSTRACT FROM AUTHOR]

Presents abstracts of several papers on vascular medicine submitted to the joint meeting of the Benelux Society for Microcirculation and the Gesellschaft für Mikrozirkulation e.V. in Mainz, Germany from October 23 to October 25, 1997. "Changes in Morphology, Solubility, and Gene Expression of Cadherins and Catenins in Shear-Stress-Exposed Endothelial Cells," by H. Schnittler, U. Ströher, U. Gotsch, D. Vestweber and H. Feldmann; "Shear-Stress Induced Endothelial NO Synthase Expression Is Dependent on a Specific Laminin Cell Interaction via the 67-kD Receptor," by T. Gloe , S. Wawro and U. Pohl; "Stretch-Induced Alterations of Endothelin Gene Expression in Native and Cultured Smooth Muscle Cells," by M. Cattaruzza, M. Lauth, C. Dimigen and M. Hecker.

The table of contents for the November 2012 issue of "Cells Tissues Organs" is presented.

Public health practice will not be able in the 21st century to ignore the impact of genomics, cell and molecular biology. It will need to take into consideration issues that include, among others: the complementary nature of social and biological models of disease, genetic exceptionalism, the readiness of public and patient to respond to genomic information, the relationship between individuals and populations, and concepts of population stratification. Health systems will need to adapt their practice and organisation to include new sequencing technologies, bioinformatic expertise and proper evaluation of genetic and molecular tests. Links with the commercial sector will increase in importance. The impact on developing countries cannot be ignored and will require special attention. Copyright © 2012 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

The table of contents for the November 2011 issue of "Cells Tissues Organs" is presented.

The table of contents for the May 2010 issue of "Cells Tissues Organs," is presented.

Presents an introduction to papers related to cellular biology that appeared in the 2005 issue of the journal "Cells Tissues Organs."

Telomeric repeat sequences, located at the end of eukaryotic chromosomes, have been detected at intrachromosomal locations in many species. Large blocks of telomeric sequences are located near the centromeres in hamster cells, and have been reported to break spontaneously or after exposure to ionizing radiation, leading to chromosome aberrations. In human cells, interstitial telomeric sequences (ITS) can be composed of short tracts of telomeric repeats (less than twenty), or of longer stretches of exact and degenerated hexanucleotides, mainly localized at subtelomeres. In this paper, we analyzed the radiation sensitivity of a naturally occurring short ITS localized in 2q31 and we found that this region is not a hot spot of radiation-induced chromosome breaks. We then selected a human cell line in which approximately 800 bp of telomeric DNA had been introduced by transfection into an internal euchromatic chromosomal region in chromosome 4q. In parallel, a cell line containing the plasmid without telomeric sequences was also analyzed. Both regions containing the transfected plasmids showed a higher frequency of radiation-induced breaks than expected, indicating that the instability of the regions containing the transfected sequences is not due to the presence of telomeric sequences. Taken together, our data show that ITS themselves do not enhance the formation of radiation-induced chromosome rearrangements in these human cell lines. Copyright © 2003 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Ectopic expression of telomerase results in an immortal phenotype in various types of normal cells, including primary human fibroblasts. In addition to its role in telomere lengthening, telomerase has now been found to have various functions, including the control of DNA repair, chromatin modification, and the control of expression of genes involved in cell cycle regulation. The investigations on the long-term effects of telomerase expression in normal human fibroblast highlighted that these cells show low frequencies of chromosomal aberrations. In this paper, we describe the karyotypic stability of human fibroblasts immortalized by expression of hTERT. The ectopic overexpression of telomerase is associated with unusual spontaneous as well as radiation-induced chromosome stability. In addition, we found that irradiation did not enhance plasmid integration in cells expressing hTERT, as has been reported for other cell types. Long-term studies illustrated that human fibroblasts immortalized by telomerase show an unusual stability for chromosomes and for plasmid integration sites, both with and without exposure to ionizing radiation. These results confirm a role for telomerase in genome stabilisation by a telomere-independent mechanism and point to the possibility for utilizing hTERT-immortalized normal human cells for the study of gene targeting. Copyright © 2003 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

In a recent paper we reported the results of an experiment carried out by analysing chromosomal damage in Chinese hamster (CHO) cells exposed to low doses of X-rays. The present investigation was undertaken in order to validate those results using a different approach, the single cell gel electrophoresis assay (comet assay) immediately after irradiation. Cells were cultured during 14 cycles, irradiation treatment was performed once per cycle when the cells were at 90–95% of confluence. Doses of 2.5, 5.0 and 10.0 mSv were used. Sequential irradiation of CHO cells induced a decrease of cells without migration and an increase of cells showing DNA damage with the three doses employed. Significant increases of low-level damaged cells (p < 0.001) were found for the 14 exposures when compared to controls except for the first irradiations with 2.5 and 10 mSv, respectively. No significant increase of the frequency of cells with severe damage was observed in any case. These findings could be explained by assuming a complex interactive process of cell recovery, DNA damage and repair together with the induction of genomic instability, the incidence of bystander effects as well as some kind of radioadaptative response of the cells. If these phenomena are limited to the cell line employed deserves further investigation. Copyright © 2003 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Hepatic progenitor cells, capable of maturing into hepatocytes and biliary cells, are hypothesized to be involved in all forms of liver regeneration and may prove clinically useful at reconstituting damaged livers. A murine hepatic progenitor cell population from young adult liver tissue has been isolated and characterized to establish a model for the development of liver cell therapies and for analysis of immune responses after transplantation. Hepatic progenitor cells were isolated from 3- to 6-week-old C57BL/6 mice using modifications of a two-stage liver perfusion technique followed by low speed centrifugation. Cellular analysis by phase contrast, fluorescent and confocal microscopy demonstrated that the hepatic progenitors (1) formed ex vivo colonies with a morphological appearance similar to committed hepatocytic progenitors isolated from embryonic mice and rats; (2) they are smaller than mature hepatocytes; (3) in culture they demonstrated peak expression of an oval cell marker at day 14, whereas albumin expression continued to increase beyond day 21 of culture, and (4) a subset of the progenitors phenotypically differentiated into mature hepatocytes or biliary cells. The unique antigenic profile of these hepatic progenitor cells and their ability to differentiate suggests that purification of the cells should allow for their potential use in transplantation.Copyright © 2003 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Background: Fibrotic replacement of the exocrine pancreatic parenchyma and infiltration of inflammatory cells are the main characteristics of chronic pancreatitis (CP). Inflammation involves prostaglandin production in numerous inflammatory and noninflammatory cells, and cyclooxygenase 2 (COX-2) is the dominant regulator of prostaglandin synthesis. Aims: In the present study, we analyzed the expression of COX-2, the key enzyme for prostaglandin synthesis in pancreatic tissues, and evaluated its relation to exocrine and endocrine tissue alterations in CP. Patients and Methods: Pancreatic tissue specimens from 27 patients undergoing pancreatic head resection for CP were included in the study. Pancreatic tissues from 14 organ donors served as controls. The tissue specimens were analyzed histopathologically and for COX-2 immunoreactivity. Results: In normal pancreatic tissue samples, COX-2 immunoreactivity was restricted to islet cells. In contrast, in early-stage CP, islets as well as ductal cells showed intense COX-2 immunoreactivity. In advanced-stage CP, ductal cells were still strongly positive for COX-2, yet islets displayed a variable COX-2 staining pattern which was associated with the distribution of insulin-positive cells and with the clinical diabetes mellitus status of the patient. Thus, patients with normal or latent diabetes mellitus status showed COX-2 immunoreactivity, whereas in diabetic patients the COX-2 immunoreactivity was decreased or absent in pancreatic islets. Conclusion: The presence of COX-2 in ductal cells of early and advanced CP, the relationship between COX-2 and insulin expression in the islets, and the diabetes mellitus status of CP patients suggest that this enzyme plays a role in the pathogenesis of exocrine and endocrine damage in CP.Copyright © 2002 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Blood-derived autografts from patients with non-Hodgkin’s lymphoma (NHL) are frequently contaminated with clonogenic lymphoma cells. In order to obtain a more efficient lymphoma cell depletion from the transplants we compared the efficiency of different purging techniques: B-cell-directed depletion, CD34+ selection by immunomagnetic beads (IMB selection) or by immunoadsorption with a biotin-avidin column (BAC selection). Furthermore, two combination approaches were investigated: IMB selection after B-cell depletion as well as BAC selection after B-cell depletion. To assess purging efficiency, fluorescence-tagged (PKH26) K422 follicular NHL cells were admixed to the respective samples of leukapheresis products. BAC selection following B-cell depletion compared to BAC selection alone showed no significant differences in CD34+ purity (77 ± 12.5% vs. 70.7 ± 5.4%) (mean ± SE) or CD34+ recovery (35.3 ± 8.5% vs. 32.8 ± 10.4%), but a significantly (p < 0.005) higher lymphoma cell purging efficiency (log 4.32 ± 0.15 vs. log 2.92 ± 0.19). IMB selection following B-cell depletion and IMB selection alone resulted in a CD34+ purity of 40.8 ± 8.7% and 64.7 ± 7.9%, a CD34+ recovery of 47.2 ± 8.3% and 26.5 ± 6.5% and a lymphoma cell purging efficiency of log 3.68 ± 0.16 and log 3.48 ± 0.21, respectively. Lymphoma cell purging efficiency after either CD34 selection method alone as well as after either double purging method was significantly higher (p < 0.0005 and p < 0.00005, respectively) compared to B-cell depletion alone (log 1.44 ± 0.23). Our results argue for the combination of different purging modalities to achieve a maximal lymphoma cell depletion. [ABSTRACT FROM AUTHOR]

Introduces chronicle of original research concerning repair and regeneration of oral and craniofacial tissues.

Presents a name list of reviewers being acknowledged by the editors-in-chief of the "Cells Tissues Organs" journal, for their comments and criticisms of the papers presented in the journal.

Common callouses are formed by the accumulation of keratinocytes in the stratum corneum in response to excess pressure or friction. We report 2 cases of unusual callous formation and an additional 25 more sequential cases that were due to excessive cell phone grip. [ABSTRACT FROM AUTHOR]

Interleukin (IL)-6 in the hypothalamus and hindbrain is an important downstream mediator of suppression of body weight and food intake by glucagon-like peptide-1 (GLP-1) receptor stimulation. CNS GLP-1 is produced almost exclusively in prepro-glucagon neurons in the nucleus of the solitary tract. These neurons innervate energy balance-regulating areas, such as the external lateral parabrachial nucleus (PBNel); essential for induction of anorexia. Using a validated novel IL-6-reporter mouse strain, we investigated the interactions in PBNel between GLP-1, IL-6, and calcitonin gene-related peptide (CGRP, a well-known mediator of anorexia). We show that PBNel GLP-1R-containing cells highly (to about 80%) overlap with IL-6-containing cells on both protein and mRNA level. Intraperitoneal administration of a GLP-1 analogue exendin-4 to mice increased the proportion of IL-6-containing cells in PBNel 3-fold, while there was no effect in the rest of the lateral parabrachial nucleus. In contrast, injections of an anorexigenic peptide growth and differentiation factor 15 (GDF15) markedly increased the proportion of CGRP-containing cells, while IL-6-containing cells were not affected. In summary, GLP-1R are found on IL-6-producing cells in PBNel, and GLP-1R stimulation leads to an increase in the proportion of cells with IL-6-reporter fluorescence, supporting IL-6 mediation of GLP-1 effects on energy balance. [ABSTRACT FROM AUTHOR]

The front cover of the journal is presented along with the issue's editors and information on membership and subscription.

The article offers information on the journal "Cells Tissues Organs," which includes topics on analyzing P-Type ATPases in major prokaryotic phyla, discussions on the journal's aims and scope, and essays on biology.

Presents a table of contents for the volume 164, 1999 issue of the journal "Cells Tissues Organs."

Background/Aims: Adipose-derived Stem Cells (ASCs) are used in Regenerative Medicine, including fat grafting, recovery from local tissue ischemia and scar remodeling. The aim of this study was to evaluate hyaluronan based gel effects on ASCs differentiation and proliferation. Methods: Comparative analyses using high (H) and low (L) molecular weight hyaluronans (HA), hyaluronan hybrid cooperative complexes (HCCs), and high and medium cross-linked hyaluronan based dermal fillers were performed. Human ASCs were characterized by flow cytometry using CD90, CD34, CD105, CD29, CD31, CD45 and CD14 markers. Then, cells were treated for 7, 14 and 21 days with hyaluronans. Adipogenic differentiation was evaluated using Oil red-O staining and expression of leptin, PPAR-γ, LPL and adiponectin using qRT-PCR. Adiponectin was analyzed by immunofluorescence, PPAR-γ and adiponectin were analyzed using western blotting. ELISA assays for adiponectin and leptin were performed. Results: HCCs highly affected ASCs differentiation by up-regulating adipogenic genes and related proteins, that were also secreted in the culture medium. H-HA and L-HA induced a lower level of ASCs differentiation. Conclusion: HCCs-based formulations clearly enhance adipogenic differentiation and proliferation, when compared with linear HA and cross-linked hyaluronans. Injection of HCCs in subdermal fat compartment may recruit and differentiate stem cells in adipocytes, and considerably improving fat tissue renewal. [ABSTRACT FROM AUTHOR]

Presents the table of contents of Volume 167, 2000 issue of the "Cells Tissues Organs" journal.

Cochlear implant stimulation creates a reduction in electrode impedance that returns to pre-stimulation levels following cessation of stimulation and is presumed to be associated with the fibrous tissue covering over the electrode array. This study assessed the possibility that transitory impedance reduction originates from a change in the membrane permeability of cells on the electrode (electropermeabilization). These changes can be recorded using the dye propidium iodide, which fluoresces upon entry into the leaky cell. The in vitro model used showed impedance reduction and dye uptake into adherent cells overlying planar gold electrodes stimulated with as little as 5 min of clinically relevant cochlear implant stimulation. The delayed additions of propidium iodide showed a similar dye uptake to those groups with concurrent dye addition, suggesting the electropermeabilization was not reversible. Further understanding of the mechanisms behind these impedance and cell permeability changes with cochlear implant electrical stimulation may provide opportunities for creating long-lasting reductions in electrode impedance. © 2014 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

The front and back covers of the journal are presented along with the issue's editors and information on topics of special interest, types of papers, and manuscript submission.

Background: The treatment of polymicrobial biofilms with antimicrobial compounds results in not only an overall loss of viability, but also compositional shifts. While DNA-based technologies may be more appropriate for the assessment of bacterial composition than culturing, these techniques amplify DNA from both live and dead cells. Propidium monoazide (PMA) has been used to discriminate between live and dead cells by blocking the DNA from membrane-damaged cells from being amplified. Aim: This study evaluated the use of PMA in a saliva-derived polymicrobial biofilm model subjected to a single chlorhexidine (CHX) treatment. Materials and Methods: The effects of PMA on viable cells were tested using both untreated and PMA-treated saliva as an inoculum. Viability was determined by plate counts, metabolic activity was determined by lactic acid production, and biofilm composition was assessed by 16S rRNA gene amplicon sequencing. Results: Exposure to a 0.2% CHX rinse (meridol® perio) reduced the viability and metabolic activity of 48-hour biofilms. The shift in biofilm composition observed after the CHX exposure was enhanced after a post-rinse PMA treatment. PMA treatment had a small effect on the measured composition of water-rinsed biofilms. Treating saliva with PMA reduced bacterial viability and shifted the bacterial composition of saliva and saliva-derived biofilms. Conclusion: The removal of DNA from non-viable cells with PMA treatment was shown to elicit an improvement in the detection of shifts in in vitro polymicrobial biofilms after antimicrobial treatment. However, PMA also influenced the ability of cells to grow, indicating that PMA should be used with caution. © 2014 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Coral reef fish present the human observer with an array of bold and contrasting patterns; however, the ability of such fish to perceive these patterns is largely unexamined. To understand this, the visual acuity of these animals - the degree to which they can resolve fine detail - must be ascertained. Behavioural studies are few in number and anatomical analysis has largely focused on estimates of ganglion cell density to predict the visual acuity in coral reef fish. Here, we report visual acuity measures for the triggerfish Rhinecanthus aculeatus. Acuity was first assessed using a series of behavioural paradigms and the figures were then contrasted with those obtained anatomically, based on photoreceptor and ganglion cell counts. Behavioural testing indicated an upper behavioural acuity of 1.75 cycles·degree-1, which is approximately the same level of acuity as that of the goldfish (Carassiusauratus). Anatomical estimates were then calculated from wholemount analysis of the photoreceptor layer and Nissl staining of cells within the ganglion cell layer. Both of these anatomical measures gave estimates that were substantially larger (7.75 and 3.4 cycles·degree-1 for the photoreceptor cells and ganglion cells, respectively) than the level of acuity indicated by the behavioural tests. This indicates that in this teleost species spatial resolution is poor compared to humans (30-70 cycles·degree-1) and it is also not well indicated by anatomical estimates. © 2014 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Background: Amniotic fluid (AF) cell populations will be applied in perinatology. We aimed to test the feasibility of large-scale cell expansion. Study Methods: We determined the best out of three published expansion protocols for mesenchymal progenitors (AF samples, n = 4) in terms of self-renewal ability. Characterization was performed based on morphology, surface marker analysis, cytogenetic stability, and differentiation potential. The conditions for the best self-renewal ability were further determined in a consecutive series (n = 159). Results: The medium containing fetal bovine serum (FBS), epidermal growth factor, insulin, transferrin, and tri-iodothyronine, combined with seeding on gelatin-coated wells, best stimulated the growth of cells with mesenchymal features, as demonstrated by flow cytometry; however, only osteogenic differentiation was possible. Large-scale testing (n = 44) failed to confirm a robust self-renewal ability. Better results were obtained (n = 88) using optimized FBS or an increased initial cell density. Eventually over 81% of cultures continued growing after the initial medium change and had mesenchymal features but failed differentiation assays. Discussion: Routine in vitro expansion of AF-derived mesenchymal cells remains problematic. Despite an increase in successful cell cultures from 40 up to 80% using optimized serum and an increased cell density, eventually cells failed to demonstrate differentiation abilities. Routine isolation and expansion from unselected AF samples remains a challenge. © 2013 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Transforming growth factor-β (TGF-β) is implicated in the regulatory expression of chemokines that control multiple steps in myogenesis. However, it remains to be established whether myostatin, a member of the TGF-β superfamily, affects chemokine expression in skeletal muscle. We investigated the effects of myostatin on the expression of mRNAs and proteins for 4 chemokines (CXCL1, CXCL2, CXCL6, CCL2) in intact and regenerating musculus longissimus thoracis from normal-muscled (NM) and double-muscled (DM) cattle. These chemokines were expressed in regenerating muscle, and their expression was always lower in DM than in NM cattle. Immunohistochemistry revealed that CXCL1 and CXCL6 were detected in the regenerating areas of myoblasts and myotubes in both NM and DM cattle. In cultures of myoblasts isolated from the regenerating muscles, significantly less CXCL1, CXCL2 and CCL2 mRNA was expressed in DM myoblasts than in NM myoblasts during the proliferating stage (P-stage). The expression of CXCL1, CXCL2 and CCL2 mRNAs in NM myoblasts and CXCL1, CXCL2 and CXCL6 mRNAs in DM myoblasts decreased upon switching from P-stage to fusion stage (F-stage). Also, the expression of CXCL1, CXCL2 and CXCL6 mRNAs was significantly lower in DM than in NM myoblasts during the F-stage. The addition of 100 ng/ml myostatin during the F-stage attenuated the expression of CXCL1 and CXCL2 mRNAs and augmented that of CCL2. These results show for the first time that myostatin regulates the differential expression of chemokines in skeletal muscle cells. Copyright © 2013 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Background/Aims: To determine the effect of arterial blood pressure (BP) reduction on endocan and soluble cell adhesion molecules' (sCAM) plasma concentration and expression of their ligands on circulatory leukocyte subpopulations. Methods: 24 hypertensive subjects of both sexes (age: 53±8 yrs) were treated with Ca-channel blocker, amlodipin (5-10 mg/day for 8 weeks; to reach BP≤139/89mmHg). The serum sCAMs and endocan concentrations were determined by ELISA kits. Level of ICAM/VCAM ligands on leukocytes was assessed by flow cytometry. Paired t-test, or t-test were used as appropriate, with Pearson's correlation calculated; p<0.05 was considered significant (SigmaPlot v.11). Results: sICAM-1 and sVCAM-1 were decreased (p≤0.001 and p=0.002, respectively), while E-selectin concentration was increased after amlodipin treatment (P=0.014). CD11a/LFA-1 (ICAM-1 and endocan ligand) was significantly increased in all three cell types with BP decrease. CD15 and CD49d/VLA-4 (VCAM-1 ligand) did not change after the treatment. There was significant positive correlation of systolic and diastolic BP with ICAM-1 and VCAM-1, and significant negative correlation of systolic BP with CD11a/LFA-1. Endocan significantly positively correlated with ICAM-1. Conclusions: The increased expression of ICAM/VACM ligands, together with decrease of sCAMs and endocan suggests the de-activation of endothelium with reduction in BP, decreasing the adherence of circulatory leukocytes to endothelium; subsequently decreasing the risk for development of atherosclerosis. Copyright © 2013 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Background: Alzheimer's disease (AD) is a frequent form of senile dementia. Neuroglobin (Ngb) has a neuroprotective role and decreases Aβ peptide levels. Ngb, promoting Akt phosphorylation, activates cell survival involving cyclic-nucleotide response element-binding protein (CREB). A new molecule (IBU-LA) was synthetized and administered to an AD rat model to counteract AD progression. Objective: The aim of this study was to investigate the IBU-LA-mediated induction of Ngb neuroprotective and antiapoptotic activities. Methods: Brain morphology was analyzed through Bielschowsky staining, Aβ(1-40) and Ngb expression by immunohistochemistry. Akt, p-Akt, CREB and p-CREB expression was evaluated by Western blot, apoptosis through cytochrome C/Apaf 1 immunocomplex formation, and TUNEL analysis. Results: Bielschowsky staining and Aβ(1-40) expression show few nerve connections and Aβ(1-40) expression in an Aβ sample, preserved neuronal cells and Aβ(1-40) expression lowering in an IBU sample, mostly in IBU-LA. The Ngb level decreases in Aβ samples, compared to control and IBU-LA samples. p-Akt/Akt and p-CREB/CREB ratios reveal a reduction in Aβ sample, going back to the basal level in control and IBU-LA samples. Cytochrome C/Apaf 1 co-immunoprecipitate occurs and TUNEL-positive nuclei percentage decreases in Aβ sample. Probe test performance shows an increased spatial reference memory in the IBU-LA compared to the Aβ sample; no significant differences were seen between the IBU-LA and IBU samples. Conclusion: This evidence reveals that IBU-LA administration has the capability to maintain a high Ngb level allowing Ngb to perform a neuroprotective and antiapoptotic role, representing a valid tool in the therapeutic strategy of AD progression. Copyright © 2013 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Aims: In the present study, we aimed to identify dietary factors related to insulin secretion function especially in healthy, non-obese Korean males. Methods: Data were obtained from the Korea National Health and Nutrition Examination Survey V-1 (KNHANES V-1). Nine hundred and twenty male adults aged >30 years of normal weight were included, excluding those with type 2 diabetes mellitus and liver disease. Alcohol use disorders identification test (AUDIT) score which represents alcohol use behaviors and nutrient intakes was used, and homeostasis model assessment of β-cell function (HOMA-B score) was calculated. Results: HOMA-B scorewas associated with age (p < 0.001), AUDIT score (p = 0.030), and percentage of fat from total energy intake (p = 0.002). HOMA-B scores in the problematic AUDIT group were significantly lower than those in the normal AUDIT group. In addition, HOMA-B scores in the lowest fat intake group were significantly lower than those in the medium fat intake group, but similar to those in the highest fat intake group. There was an interaction between alcohol use behaviors and percentage of fat from energy intake in determining HOMA-B score (p for interaction = 0.034). Conclusions: Alcohol use behaviors and percentage of fat from energy intake were found to be associated with HOMA-B score in healthy, non-obese Korean males. Copyright © 2013 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Introduction: Flow cytometry analysis of lymphocyte subsets in peripheral blood is a common technique in diagnostic laboratories. Abnormal values have been identified in prevalent infections, autoimmune disorders and neoplastic diseases. Reference ranges for lymphocyte subsets of a healthy population from Spain are scarce. Methods: The study was performed on 319 healthy subjects, aged 4-88 years, from 709 individuals enrolled in the GAIT-2 Project (Genetic Analysis of Idiopathic Thrombophilia). Health status, age, sex, fertility, BMI and lifestyle (physical activity, cigarette smoking and ethanol intake) were assessed using standardized criteria. The percentage of lymphocyte subsets was determined using flow cytometry (Lymphogram™). Percentages of CD3+, CD4+, CD8+, CD19+, CD3-CD56+, CD3+CD4-CD8- double-negative (DN) T cells, CD3+CD4+CD8+ double-positive T cells and the CD4+/CD8+ ratio were recorded for each case. Results: Children had a significantly higher percentage of CD19+ and DN cells than adults. Women had a significantly higher percentage of CD3+ and CD4+ and a lower percentage of natural killer cells than men. Increases in BMI were inversely associated with the percentage of DN cells. Physical activity increased the percentage of lymphocytes and DN cells. Alcohol consumers had a lower percentage of CD19+ and DN cells, and a higher percentage of CD4+. Conclusion: This study provides reference ranges for lymphocyte subsets of healthy children and adults in a Mediterranean population (Spain) and determines the influence of lifestyle factors on these values. Copyright © 2012 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Background/Aims: SM22α, transgelin, has been revealed to be specifically expressed in glomerular epithelial cells and interstitial cells, according to the nature of the renal injury. In this study, quantitative analyses of SM22α positivity were performed to investigate the pathological significance of its expression. Methods: Kidney samples of adriamycin nephropathy underwent immunohistochemistry with a newly established anti-SM22α monoclonal antibody. The SM22α positivity was quantified by an image analyzer. The correlation of the histological values with biochemical data was investigated statistically. Microstructural localization of SM22α was studied by immunoelectron microscopy. Results: SM22α was expressed along the dense basal microfilaments of degenerating podocytes, and diffusely in interstitial cells. Both the extent and intensity of SM22α expression in glomerular and tubulointerstitial area were correlated with the deterioration of renal function and the severity of proteinuria. Stepwise multiple linear regression analysis revealed that the extent of its positivity in glomerular or tubulointerstitial area was the determinant of the amount of proteinuria or the deterioration of creatinine clearance (Ccr), respectively. Inversely, the deterioration of Ccr was the most important predictor of SM22α expression. Conclusion: SM22α expression in podocytes and interstitial cells represented the severity of proteinuria and the deterioration of renal function. SM22α expression in renal tissues might be a hallmark of kidney diseases. Copyright © 2011 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Iminodiacetic acid (IDA) is widely used as an intermediate in the manufacture of chelating agents, glyphosate herbicides and surfactants. In the current work, the fragment with the length of 1,110 bp encoding the Acidovorax facilis nitrilase was obtained. The recombinant nitrilase expressed in Escherichia coli BL21 (DE3) was successfully used in the production of IDA from iminodiacetonitrile. To improve the stability of operation, the recombinant cells were entrapped in polyvinyl alcohol (PVA) and sodium alginate (SA) copolymer. The maximum relative nitrilase activity with 98.1% was further observed at 1.0% SA, 8.0% PVA, 1.0% CaCl2, and 5.0% wet cells, under conditions of 1.0% iminodiacetonitrile in distilled water and a temperature of 40°C, respectively. The entrapped cells facilitated easy separation and good recycling compared with free cells. Moreover, the immobilized cells showed good operation and storage stability. This report is the first to describe IDA preparation using immobilized recombinant E. coli harboring nitrilase. Copyright © 2012 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Objective: To determine the antioxidant activity and cytotoxicity of Phyllanthus virgatus crude extract compared to Phyllanthus amarus.Methods: Phenolic contents of the hydromethanolic extracts were measured using Folin-Ciocalteu reagent. Antioxidant activity was evaluated by the 2,2-diphenyl-1-picrylhydrazyl hydrate free radical scavenging and antilipid peroxidation assays. Cytotoxicity on human hepatoma HepG2 cells was assessed by trypan blue and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide viability assays. A stereomicroscope was used to observe and photograph the morphology of the cells. Oxygen consumption of the HepG2 cells was measured using a Clark oxygen electrode. Results: The extract of P. virgatus, which contained more phenolic compounds than P. amarus extract, had higher cytotoxicity and showed higher free radical scavenging activity and more inhibition of peroxidation in a linoleic acid system. P. virgatus extract conspicuously increased the oxygen consumption of HepG2 cells, while P. amarus extract had little stimulatory effect. Conclusion: The hydromethanolic extract of P. virgatus had stronger antioxidant and cytotoxic action than P. amarus extract. The stimulation of HepG2 cell respiration by P. virgatus extract suggests the extract alters mitochondrial function; this action could play a role in the cytotoxicity of this plant. Copyright © 2011 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Background/Aims: Experimental studies in vitro suggest that somatostatin and some of its analogues used in clinical practice, such as octreotide, may have potent antiangiogenic properties. However, the clinical transposition of these data is difficult. Methods: To address this issue, we designed a comparative study of the effects of somatostatin and octreotide on the interactions between neoplastic endocrine cells and endothelial cells in several in vitro and in vivo experimental models, including primary cultures of human umbilical vein endothelial cells (HUVEC), indirect cocultures between HUVEC and the somatostatin-producing endocrine cell line STC-1, and an animal model of intrahepatic dissemination of STC-1 cells. Results: 10-8M octreotide markedly inhibited both basal and VEGF-stimulated HUVEC proliferation, had no effect on endothelial cell migration, but inhibited endothelial tubule formation. HUVEC cocultured with the somatostatin- and VEGF-producing STC-1 cells presented a markedly decreased proliferation, a slightly increased motility and an increased capacity of tubule formation; in this system, the inhibition of endothelial cell proliferation was abolished by neutralizing anti-somatostatin but was restored in the presence of anti-VEGF antibodies. This suggests that somatostatin is able to antagonize the effects of VEGF on endothelial cell proliferation but not on endothelial cell sprouting. Finally, no significant effect of octreotide on tumor growth and intratumoral microvascular density was detected in an experimental model of intrahepatic dissemination of STC-1 cells. Conclusion: The in vitro antiangiogenic effects of somatostatin and its analogues are likely to be efficiently counterbalanced in the tumor microenvironment by the concomitant release of proangiogenic factors like VEGF. Copyright © 2011 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Background/Aims: Cocaine- and amphetamine-regulated transcript (CART) is an anorexigenic regulatory peptide highly expressed in the brain's appetite control centers, but also in peripheral neurons and in endocrine cells in the adrenal medulla, thyroid, pancreatic islets, and in the gastrointestinal tract. Plasma levels of CART were recently shown to be elevated in patients with neuroendocrine tumors (NETs), but the cellular sources of CART in NETs have remained unknown. The aim of the study was to establish whether CART is expressed in various types of NETs and, if so, to examine the frequency, distribution and phenotype of CART-expressing cells. Methods: Tumor specimens from 133 NETs originating in the stomach, ileum, rectum, pancreas and thyroid were examined with immunohistochemistry and in situ hybridization. The expression of CART was quantified and the CART-expressing cells were phenotyped by double staining for established markers and hormones. Results: CART-expressing tumor cells were found in the majority of the examined NETs. The expression pattern of CART was highly heterogeneous not only between tumors, but also within individual tumors. In 14% of the NETs, CART was found in a major population of the tumor cells. Conclusion: CART is produced in the majority of NETs, regardless of tumor origin. This likely explains the elevated levels of circulating CART in certain NETs patients, as recently described. CART could therefore prove to be a useful tool in the diagnostics of NETs not only in blood samples, but also in histopathological specimens. Copyright © 2011 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Background/Aims: We identified carnosinase-1 (CN-1) as risk-factor for diabetic nephropathy (DN). Carnosine, the substrate for CN-1, supposedly is a protective factor regarding diabetic complications. In this study, we hypothesized that carnosine administration to diabetic rats might protect the kidneys from glomerular apoptosis and podocyte loss. Methods: We examined the effect of oral L-carnosine administration (1g/kg BW per day) on apoptosis, podocyte loss, oxidative stress, AGEs and hexosamine pathway in kidneys of streptozotocin-induced diabetic Wistar rats after 3 months of diabetes and treatment. Results: Hyperglycemia significantly reduced endogenous kidney carnosine levels. In parallel, podocyte numbers significantly decreased (-21% compared to non-diabetics, p<0.05), apoptotic glomerular cells numbers increased (32%, compared to non-diabetic, p<0.05) and protein levels of bax and cytochrome c increased (175% and 117%). Carnosine treatment restored carnosine kidney levels, prevented podocytes loss (+23% compared to diabetic, p<0.05), restrained glomerular apoptosis (-34% compared to diabetic; p<0.05) and reduced expression of bax and cytochrome c (-63% and -54% compared to diabetics, both p<0.05). In kidneys of all diabetic animals, levels of ROS, AGEs and GlcNAc-modified proteins were increased. Conclusion: By inhibition of pro-apoptotic signaling and independent of biochemical abnormalities, carnosine protects diabetic rat kidneys from apoptosis and podocyte loss. Copyright © 2011 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

The table of contents for volume 39, 2011 of the periodical "Cells Tissues Organs" is presented.

Background: Myelin-associated molecules are major impediments to axon regeneration after optic nerve injury. Intravitreal injection of α-crystallin can protect axons from optic nerve degeneration after crushing in rats. Objectives: Our purpose was to investigate whether α-crystallin could counteract the inhibitory effect of myelin and promote neurite growth. Methods: Newborn rat retinal neurons were cultured on myelin-coated dishes with DMEM containing α-crystallin (10-4 g/l) or bovine serum albumin. The density of neurons with neurites and the length of the longest neurite of the cells were analyzed on days 1, 3 and 5. Results: Cultures containing α-crystallin had significantly higher neurite-containing cell densities, and the neurites were significantly longer compared with cultures containing bovine serum albumin. These findings indicated that α-crystallin could counteract the effect of myelin inhibitory factors and stimulate neurite growth. Copyright © 2010 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Introduction: In this study, we aimed to form spheroids with the bovine placental trophoblast cell line F3. Spheroids are 3-dimensional culture models which can be used to conduct versatile in vitro and in vivo experiments. Materials and Methods: The spheroids were generated using the hanging drop technique, 25% methocel and matrigel. The F3 spheroids were characterized morphologically by light microscopy and transmission (TEM) and scanning electron microscopy (SEM) and immunohistochemistry (ezrin, vimentin, cytokeratin, placental lactogen). The fluorescent dyes calcein and ethidium homodimer were used to determine the viability of the spheroidal F3 cells by immunofluorescence microscopy. Results: The cell line F3 only formed spheroids by the hanging drop technique when matrigel was added. The trophoblast spheroids were delimited and fully covered by extracellular matrix (light microscopy/TEM/SEM). Cells contributing to spheroids could not be discriminated from each other (light microscopy). The outer spheroidal layer consisted of cells which possessed an apical pole with microvilli that were directed to the outside (light microscopy/TEM). All of the spheroidal F3 cells expressed ezrin, vimentin and cytokeratin, but not placental lactogen. The spheroid core contained degenerating cells whilst the F3 cells of the outer rim were viable (TEM/immunofluorescence microscopy). Discussion: We have established a 3-dimensional spheroid model for the bovine placental trophoblast cell line F3. The developed culture model might prove valuable for future in vitro studies on the differentiation of bovine trophoblast cells. Copyright © 2010 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Retinal topography, cell density and sizes of ganglion cells in the Caspian seal (Pusa caspica) were analyzed in retinal whole mounts stained with cresyl-violet. The topographic distribution of ganglion cells displayed an area of high cell density located in the temporal quadrant of the retina and was similar to the area centralis of terrestrial carnivores. It extended nasally, above the optic disk, as a streak of increased cell density. In different whole mounts, the peak cell density in the high-density area ranged from 1,684 to 1,844 cells/mm2 (mean 1,773 cells/mm2). The cell density data predict a retinal resolution of around 8.5 cycles/degree in water. A distinctive feature of the Caspian seal's retina is the large size of ganglion cells and the low cell density compared to terrestrial mammals. The ganglion cell diameter ranged from 10 to 58 μm. Cell size histograms featured bimodal patterns with groups of small and large ganglion cells. The large cells appeared similar to α-cells of terrestrial mammals and constituted 7% of the total ganglion cell population. Copyright © 2010 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Background and Purpose: The aims of this study were to evaluate the role of tumor necrosis factor-α (TNF-α) polymorphisms in patients susceptible to ischemic stroke and silent brain infarction (SBI), and to determine the relationship between TNF-α polymorphisms and plasma total homocysteine (tHcy) levels. Methods: We studied 237 patients with ischemic stroke, 257 patients with SBIs, and 216 control subjects. For control subjects, we selected healthy individuals matched for gender and age from those individuals who came to our hospital for health examinations. The TNF-α-308G→A and -238G→A genotypes were determined by the polymerase chain reaction-restriction fragment length polymorphism method. Results: The frequency of the TNF-α-308G→A polymorphism was significantly different between the patients with ischemic stroke and the control group (GG vs. GA+AA; adjusted odds ratio, AOR, 0.50; 95% CI 0.255-0.989). By subgroup analysis, when tHcy levels were stratified into high (>10.80 μmol/l), moderate (8.21-10.80 μmol/l), and low levels (<8.21 μmol/l), the frequency of the TNF-α-308GA+AA genotype in the highest tertile group was higher than in the lowest tertile group (AOR 2.46; 95% CI 1.063-5.699). However, the relationship between SBI susceptibility and polymorphisms of TNF-αwas not established. The tHcy levels were significantly and inversely correlated with folate levels in the TNF-α-308GG and TNF-α-238GG genotypes in the ischemic stroke, SBI, and control groups (p< 0.05). Conclusions: Our results suggest that the TNF-α-308G→A polymorphism is responsible for susceptibility to ischemic stroke and is associated with high tHcy levels in Koreans. Copyright © 2010 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Objectives: Successful engraftment of human T-lymphotropic virus type 1 (HTLV-1)-infected cells and a marked increase of proviral DNA loads (PVLs) in non-obese diabetic/severe combined immunodeficient (NOD/SCID)/γcnull (NOG) mice have been reported. Whether the increased PVL in transplanted mice is due to the new infection of HTLV-1 was examined. Methods: Mononuclear cells from 3 NOG mice with primary engraftment from asymptomatic HTLV-1 carriers were transplanted into a second group of NOG mice. HTLV-1 PVL, proviral integration by fluorescence in situ hybridization assay, expression of viral antigen, and T-cell clonality were analyzed. Results: The PVLs in the secondarily transplanted NOG mice were significantly higher than those of primarily transplanted NOG mice. Multiple signals of HTLV-1 proviruses in the nucleus of the infected cells were revealed by fluorescence in situ hybridization analysis. Expression of HTLV-1 tax/rex mRNA and antigen was observed. The variety of T-cell clones was limited in the transplanted NOG mice. Conclusions: Multiple proviral integrations were considered to be due to the new infection from HTLV-1-infected cells to the other cells. Only a certain fraction of T cells seemed to have selectively survived in NOG mice after engraftment. Copyright © 2010 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]