Isolation, Culture and Bioactive Substances Elicitation of Sanghuangporus Baumii

Sanghuangporus baumii is a forest pathogenic fungus and also a medicine mushroom. In the wild, S. baumii parasitize on living host plants and there have been no reports of cultivating fruiting bodies under artificial conditions. In this study, we identified and isolated a S. baumii strain, and successfully cultivated the fruiting bodies on sawdust medium by optimizing culture conditions. The optimum medium, culture temperature and pH for mycelial growth of S. baumii were WBA and YPA, at pH 5.5–7.5 and 28℃, respectively. The contents of total flavonoids, total polysaccharides and total triterpenoids in S. baumii were compared with those in other two medicinal sanghuang, S. vaninii and S. sanghuang. The results showed that both the fruiting bodies and mycelia of S. baumii were rich in bioactive substances. The content of total flavonoids was higher in the fruiting bodies (34.91 mg/g), while the contents of total polysaccharides and total triterpenoids were higher in the mycelia (45.41 mg/g, 14.06 mg/g, respectively). In addition, the mycelia of S. baumii could be elicited to produce more bioactive substances. The use of sealing film in culture increased the polysaccharides content in mycelia by 51.8%, while the light increased the flavonoids content by 151.0%. The cultivation of fruiting bodies and the elicitation of bioactive substances from mycelia provide biological materials for the study and utilization of S. baumii.

Development of accurate temperature regulation culture system with metallic culture vessel demonstrates different thermal cytotoxicity in cancer and normal cells

Hyperthermia has been studied as a noninvasive cancer treatment. Cancer cells show stronger thermal cytotoxicity than normal cells, which is exploited in hyperthermia. However, the absence of methods evaluating the thermal cytotoxicity in cells prevents the development of hyperthermia. To investigate the thermal cytotoxicity, culture temperature should be regulated. We, thus, developed a culture system regulating culture temperature immediately and accurately by employing metallic culture vessels. Michigan Cancer Foundation-7 cells and normal human dermal fibroblasts were used for models of cancer and normal cells. The findings showed cancer cells showed stronger thermal cytotoxicity than normal cells, which is quantitatively different from previous reports. This difference might be due to regulated culture temperature. The thermal stimulus condition (43 °C/30 min) was, further, focused for assays. The mRNA expression involving apoptosis changed dramatically in cancer cells, indicating the strong apoptotic trend. In contrast, the mRNA expression of heat shock protein (HSP) of normal cells upon the thermal stimulus was stronger than cancer cells. Furthermore, exclusively in normal cells, HSP localization to nucleus was confirmed. These movement of HSP confer thermotolerance to cells, which is consistent with the different thermal cytotoxicity between cancer and normal cells. In summary, our developed system can be used to develop hyperthermia treatment.

Effects of low temperature on electrophysiology and mechanophysiology of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs)

Studies related to low temperature and their effect on cardiomyocytes are essential as hypothermia—like situations have been known to induce arrhythmia or ventricular fibrillation. Till date, several studies have been carried out on animals and their electrophysiological responses have been studied in the form of action potential. However, for a complete assessment of the effect of low temperature, mechanophysiological changes along with electrophysiological changes need to be investigated, at the tissue level. In this study, the effect of culture temperature on cell growth has been studied by measuring the field potential and contractility of human induced pluripotent stem cell-derived cardiomyocytes. This study has the potential to further improve the understanding of low temperature on human cells.

Production of Polyhydroxyalkanoates (PHAs) by Vibrio alginolyticus Strains Isolated from Salt Fields

Vibrio alginolyticus is a halophilic organism usually found in marine environments. It has attracted attention as an opportunistic pathogen of aquatic animals and humans, but there are very few reports on polyhydroxyalkanoate (PHA) production using V. alginolyticus as the host. In this study, two V. alginolyticus strains, LHF01 and LHF02, isolated from water samples collected from salt fields were found to produce poly(3-hydroxybutyrate) (PHB) from a variety of sugars and organic acids. Glycerol was the best carbon source and yielded the highest PHB titer in both strains. Further optimization of the NaCl concentration and culture temperature improved the PHB titer from 1.87 to 5.08 g/L in V. alginolyticus LHF01. In addition, the use of propionate as a secondary carbon source resulted in the production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV). V. alginolyticus LHF01 may be a promising host for PHA production using cheap waste glycerol from biodiesel refining.

Effect of low temperature culture on the biological characteristics and aggressiveness of Sclerotinia sclerotiorum and Sclerotinia minor

Sunflower White Mold caused by Sclerotinia sclerotiorum and Sclerotinia minor is a devastating disease worldwide. To investigate the effect of low temperature (4 °C) on biological characteristics and aggressiveness of isolates of the two species, which were collected from the same field in Baiyinchagan, Inner Mongolia, their mycelial growth rate, oxalic acid secretion level and polygalacturonase activity were compared under normal culture temperature (23 °C) and low temperature (4 °C). Aggressiveness was also evaluated on detached leaves by inoculating the isolates produced in both temperatures. The results suggested that culture of isolates at 4 °C not only promoted mycelial growth, but also enhanced secretion of oxalic acid and polygalacturonase activity of both S. sclerotiorum and S. minor isolates compared to that cultured at 23 °C. Additionally, the corresponding aggressiveness of tested isolates of the two species also increased after culture at 4 °C. However, S. sclerotiorum always showed faster mycelial growth, higher oxalic acid levels and greater polygalacturonase activity than S. minor at both 23 °C and 4 °C, indicating that S. sclerotiorum is generally the more aggressive species than S. minor.

A Cell Fate Switch in the Caenorhabditis elegans Seam Cell Lineage Occurs Through Modulation of the Wnt Asymmetry Pathway in Response to Temperature Increase

Populations often display consistent developmental phenotypes across individuals despite inevitable biological stochasticity. Nevertheless, developmental robustness has limits, and systems can fail upon change in the environment or the genetic background. We use here the seam cells, a population of epidermal stem cells in Caenorhabditis elegans, to study the influence of temperature change and genetic variation on cell fate. Seam cell development has mostly been studied so far in the laboratory reference strain (N2), grown at 20° temperature. We demonstrate that an increase in culture temperature to 25° introduces variability in the wild-type seam cell lineage, with a proportion of animals showing an increase in seam cell number. We map this increase to lineage-specific symmetrization events of normally asymmetric cell divisions at the fourth larval stage, leading to the retention of seam cell fate in both daughter cells. Using genetics and single-molecule imaging, we demonstrate that this symmetrization occurs via changes in the Wnt asymmetry pathway, leading to aberrant Wnt target activation in anterior cell daughters. We find that intrinsic differences in the Wnt asymmetry pathway already exist between seam cells at 20° and this may sensitize cells toward a cell fate switch at increased temperature. Finally, we demonstrate that wild isolates of C. elegans display variation in seam cell sensitivity to increased culture temperature, although their average seam cell number is comparable at 20°. Our results highlight how temperature can modulate cell fate decisions in an invertebrate model of stem cell patterning.

Cell adhesion control by photoinduced LCST shift of PNIPAAm-based brush scaffolds

Cell adhesion was markedly regulated on spiropyran-containing PNIPAAm-based brush scaffolds by photoinduced LCST shift across a standard culture temperature.