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Coagulation factor XII precursor (EC (Hageman factor) (HAF) [Contains: Coagulation factor XIIa heavy chain; Beta-factor XIIa part 1; Coagulation factor XIIa light chain (Beta-factor XIIa part 2)]


Multigenerational effects of carbendazim in Daphnia magna: From a subcellular to a population level.

Anthropogenic activities such as the use of pesticides may affect aquatic biota populations, due to potential agricultural runoffs or disposals. Carbendazim is one example of a widely used fungicide with a high potential to end up in aquatic ecosystems through runoff. Deleterious effects observed at the individual level are possibly explained by changes in homeostasis at the cellular level, and both factors can then be used to predict effects at the population level. In the present study, an isoclonal population of Daphnia magna (clone K6) was exposed to a concentration that mimics relevant levels of carbendazim in the environment over 12 generations. The effects of carbendazim were assessed in some generations using the following endpoints: biochemical biomarkers (cholinesterase, catalase, and glutathione-S-transferase), lipid peroxidation and energy-related parameters (carbohydrates, lipids, and proteins along with available energy and energy consumption), parental longevity, and population growth (r). Long-term exposure to carbendazim had no effect on the intrinsic rate of natural increase (r) of adult D. magna, but longevity was decreased at the F12 generation compared to that of control. Differences between the exposed and nonexposed populations were found for cholinesterase, glutathione-S-transferase, and lipid peroxidation. However, for catalase and energy-related parameters, no differences were observed between these 2 populations. Natural variability was seen throughout the test period, under control conditions, within the 12 generations. Overall, carbendazim induced some effects at the subcellular level that translated into changes in longevity but these later vanished in terms of population effects. Environ Toxicol Chem 2019;38:412-422. © 2018 SETAC.

MeSH Terms

  • Animals
  • Benzimidazoles
  • Carbamates
  • Daphnia
  • Ecosystem
  • Fungicides, Industrial
  • Lipid Peroxidation
  • Longevity
  • Reproduction
  • Water Pollutants, Chemical


  • Biochemical biomarkers
  • Carbendazim
  • Daphnia magna
  • Energy reserves
  • Multigenerations

In vitro cytotoxicity of maxillofacial silicone elastomers: effect of accelerated aging.

The purpose of this in vitro study was to evaluate the cytotoxicity of three maxillofacial silicone elastomers at 24, 48, and 72 h on L-929 cells and to determine the effect of accelerated aging on the cytotoxicity of these silicone elastomers. Disc-shaped test samples of maxillofacial silicone elastomers (Cosmesil, Episil, Multisil) were fabricated according to manufacturers' instructions under aseptic conditions. Samples were then divided into three groups: (1) not aged; (2) aged for 150 h with an accelerated weathering tester; and (3) aged for 300 h. Then the samples were placed in Dulbecco's Modified Eagle Medium/Ham's F12 (DMEM/F12) for 24, 48, and 72 h. After the incubation periods, cytotoxicity of the extracts to cultured fibroblasts (L-929) was measured by MTT assay. The degree of cytotoxicity of each sample was determined according to the reference value represented by the cells with a control (culture without sample). Statistical significance was determined by repeated measurement ANOVA (p < 0.01) followed by Duncan's test (p < 0.05). All test materials in each group demonstrated high survival rates in MTT assay (Episil; 93.84%, Multisil; 88.30%, Cosmesil; 87.50%, respectively); however, in all groups, Episil material demonstrated significantly higher cell survival rate after each of the experimental incubation periods (p < 0.05). Accelerated aging for 150 and 300 h had no significant effect on the biocompatibility of maxillofacial silicone elastomers tested (p > 0.05).

MeSH Terms

  • Aging
  • Animals
  • Biocompatible Materials
  • Cell Line
  • Cell Survival
  • Humans
  • Materials Testing
  • Maxillofacial Prosthesis
  • Mice
  • Silicone Elastomers

In vitro cartilage regeneration from proliferated adult elastic chondrocytes.

The purpose of this study was to investigate cellular feasibility in the proliferation and differentiation status of adult chondrocytes for cartilage regeneration in comparison to fetal chondrocytes. Primary cells were isolated from adult (n = 6) and fetal (n = 6) sheep ear cartilages and expanded in 10% fetal bovine serum (FBS) containing Ham's F12 medium, in which adult and fetal cell proliferation rates were compared using a WST-1 assay kit. Approximately 4 million cells were seeded onto each 1 x 1 x 0.2-cm (200 microL) nonwoven fabric scaffold made from polyglycolic acid. Cell/polymer constructs were cultured in serum-free DMEM/F12 medium supplemented with 5 ng/mL TGF-beta2 and 5 ng/mL des(1-3)IGF-I (adult chondrocytes, group A) or in 10% FBS containing Ham's F12 medium (adult chondrocytes, group B, and fetal chondrocytes, group C) as controls in a rotating bioreactor for 6 weeks. The proliferation assay showed that fetal cells had a significantly better growth potential than did adult cells. Histology and extracellular matrix analyses revealed that groups A and C qualitatively displayed better matrix deposition than did group B. In conclusion, although adult sheep elastic chondrocytes had less growth potential than did fetal cells, the serum-free medium supplemented with growth factors significantly enhanced the production of cartilage matrix secreted from proliferated adult sheep elastic chondrocytes.

MeSH Terms

  • Age Factors
  • Aging
  • Animals
  • Cartilage
  • Cell Proliferation
  • Cell Separation
  • Chondrocytes
  • DNA
  • Ear
  • Elastic Tissue
  • In Vitro Techniques
  • Regeneration
  • Sheep
  • Tissue and Organ Harvesting
  • Trinucleotide Repeat Expansion

Expression of angiotensin II AT2 receptor mRNA during development of rat kidney and adrenal gland.

The angiotensin II (ANG II) receptors have been pharmacologically classified into two major distinct types, designated AT1 and AT2. A high transient expression of AT2 receptors in the fetal tissues has been previously demonstrated. This study describes the cellular distribution of AT2 receptor mRNA in the developing rat kidney and adrenal gland by in situ hybridization with 35S-labeled cRNA probes. From day 12 of fetal life (F12) to day 15 postpartum (D15) AT2 mRNA was detected in the undifferentiated nephrogenic mesenchymal tissue but not in the immature and mature glomeruli and tubules of the kidney. No AT2 mRNA was observed in the kidney after D22. The adrenal gland also expressed AT2 receptor mRNA early during development from F12 but, unlike the kidney, continuously expressed the mRNA at high levels through to adulthood. The disappearance of AT2 mRNA in the kidney was synchronous with the completion of nephrogenesis and suggests that ANG II might act through this receptor as a differentiation/growth factor during nephron development. In the adrenal gland ANG II could act as a hormone and also as a differentiation/growth factor via the AT2 receptor.

MeSH Terms

  • Adrenal Glands
  • Aging
  • Animals
  • Animals, Newborn
  • Embryonic and Fetal Development
  • Fetus
  • Kidney
  • RNA, Messenger
  • Rats
  • Rats, Sprague-Dawley
  • Receptors, Angiotensin

Effect of age on the differentiation of porcine adipose stromal-vascular cells in culture.

Stromal-vascular (S-V) cells isolated from adipose tissue of newborn pigs (NBPC) and mature pigs (MPC) by collagenase digestion were used to evaluate differences in preadipocyte culture and development. Cells were seeded at a density of 3 x 10(4) cells/cm2 on six-well (35-mm) tissue culture plates in 3 mL of DMEM/HAM's F12 medium plus 10% fetal calf serum and cultured at 37 degrees C under a humidified atmosphere of 95% air:5% CO2 for 24 h. Cells were then washed thoroughly in DMEM/HAM's F12 medium without fetal calf serum and maintained in serum free (SF) medium or SF medium supplemented with 2.5% newborn pig serum (NBPS) or mature pig serum (MPS) for 12 d. After 1 d, more NBPC adhered to the culture plates, as indicated by DNA values. After 12 d, protein per culture well was not significantly different, but DNA concentration per well remained higher (P < .05) in cultures of NBPC than in the MPC cultured in the same medium, indicating fewer MPC. Protein:DNA ratios were higher (P < .05) in cultures of MPC regardless of the medium, reflecting larger cell size. More cells containing fat deposits were seen with NBPC in all conditions in comparison with MPC, and more fat was deposited in NBPC in SF than in SF plus NBPS or MPS. The NBPC had higher (P < .05) sn-glycerol-3-phosphate dehydrogenase (GPDH; EC per protein than MPC regardless of the medium. For both cell types, GPDH activity in either serum was less than activity of cells grown in SF.(ABSTRACT TRUNCATED AT 250 WORDS)

MeSH Terms

  • Adipocytes
  • Aging
  • Animals
  • Cell Count
  • Cell Differentiation
  • Cell Division
  • Cells, Cultured
  • DNA
  • Male
  • Stromal Cells

Mitogen induced proliferation of isolated adult mouse Schwann cells.

The proliferation of neonatal Schwann cells (SCs) in response to mitogenic agents has been well analyzed in vitro, but a limited range of mitogens have been defined. We investigated whether three identified neonatal SC mitogens [glial growth factor (GGF), platelet-derived growth factor BB (PDGF-BB), and basic fibroblast growth factor (bFGF)] are required to stimulate mitosis of adult SCs. Adult SCs were isolated from mouse sciatic nerves by mechanical and chemical dissociation, following three experimental steps: 1) culturing the dissociated cells for 24 hr in 10% FCS-F12 medium, 2) culturing these cells in serum-free medium for the next 48 hr, and 3) purifying adult SCs by differential adhesion. We describe a new method for preparation of SCs from peripheral nerves of adult mouse that provides 99.5% pure SCs populations at cell yields of greater than 3 x 10(3) cells/mg of starting nerve wet weight within 5 culture days. Although mitosis of SCs in culture in response to mitogens requires the presence of serum, the complex nature of serum renders difficult a complete analysis of mitogens required for SCs DNA synthesis, so we examined the proliferating response of adult SCs to GGF, PDGF-BB, and bFGF in serum-free medium. GGF alone had mitogenicity for adult SCs in a dose-dependent manner, and synergistic activation coupling with forskolin was not observed. Neither PDGF-BB nor bFGF was mitogenic for adult SCs when used alone or with forskolin.(ABSTRACT TRUNCATED AT 250 WORDS)

MeSH Terms

  • Aging
  • Animals
  • Cell Division
  • Cell Separation
  • Cells, Cultured
  • Culture Media, Serum-Free
  • Cyclic AMP
  • Immunohistochemistry
  • Intracellular Membranes
  • Mice
  • Mice, Inbred C57BL
  • Mitogens
  • Schwann Cells

Isolation, characterization, and in vitro culture of larval and adult epidermal cells of the frog Xenopus laevis.

Methods for the isolation and in vitro culture of larval and adult Xenopus laevis epidermal cells have been developed. Epidermal cells of stage 52-54 tadpoles and adult epidermal cells were enzymatically dissociated and purified (98%) by Percoll-density centrifugation and unit-gravity sedimentation. Both cell types attached on fibronectin-coated dishes and proliferated for 1 wk when the proper medium was used. There were four significant differences between larval and adult cells: a) Adult cells had a greater buoyant density than larval cells. b) Keratin synthesis patterns were markedly different. c) A combination of medium F12 and Eagle's minimum essential medium was optimal for growth of larval cells whereas MCDB151 medium was optimal for adult cells. d) Adult cells needed fetal bovine serum (greater than 5%) whereas larval cells grew without fetal bovine serum. In contrast to these differences, larval and adult cells had two similar properties: a) Insulin had a potent effect on the growth of both cells, and b) The optimal Ca concentration for cell growth was quite low for both cell types; 0.1 mM for larval cells and below 0.05 mM for adult cells. These results suggest that low Ca levels are essential for both cornifying (adult) and uncornifying (larval) amphibian keratinocytes. The culture techniques described herein for larval and adult epidermal cells provide a new in vitro model for analyzing development of the epidermis during amphibian metamorphosis.

MeSH Terms

  • Aging
  • Animals
  • Blood
  • Calcium
  • Cell Division
  • Cell Separation
  • Cells, Cultured
  • Centrifugation, Density Gradient
  • Culture Media
  • Epidermal Cells
  • Epidermis
  • Fibronectins
  • Growth Substances
  • Insulin
  • Keratins
  • Larva
  • Metamorphosis, Biological
  • Xenopus laevis