Asexual development (conidiation) of the filamentous fungus Aspergillus nidulans occurs via balanced activities of multiple positive and negative regulators. For instance, FluG (+) and SfgA (2) govern upstream regulation of the developmental switch, and BrlA (+) and VosA (2) control the progression and completion of conidiation. To identify negative regulators of conidiation downstream of FluG-SfgA, we carried out multicopy genetic screens using sfgA deletion strains. After visually screening .100,000 colonies, we isolated 61 transformants exhibiting reduced conidiation. Responsible genes were identified as AN3152 (nsdD), AN7507, AN2009, AN1652, AN5833, and AN9141. Importantly, nsdD, a key activator of sexual reproduction, was present in 10 independent transformants. Furthermore, deletion, overexpression, and double-mutant analyses of individual genes have led to the conclusion that, of the six genes, only nsdD functions in the FluG-activated conidiation pathway. The deletion of nsdD bypassed the need for fluG and flbA$flbE, but not brlA or abaA, in conidiation, and partially restored production of the mycotoxin sterigmatocystin (ST) in the DfluG, DflbA, and DflbB mutants, suggesting that NsdD is positioned between FLBs and BrlA in A. nidulans. Nullifying nsdD caused formation of conidiophores in liquid submerged cultures, where wild-type strains do not develop. Moreover, the removal of both nsdD and vosA resulted in even more abundant development of conidiophores in liquid submerged cultures and high-level accumulation of brlA messenger (m)RNA even at 16 hr of vegetative growth. Collectively, NsdD is a key negative regulator of conidiation and likely exerts its repressive role via downregulating brlA.T HE filamentous ascomycete Aspergillus nidulans has served as an excellent model system for studying cell biology, asexual development (conidiation), and secondary metabolism (Timberlake 1990;Martinelli 1994;Yu and Keller 2005). The A. nidulans asexual reproductive cycle can be divided into two distinct phases: growth and development. The growth phase involves germination of an asexually derived spore called a conidium and formation of an undifferentiated network of interconnected hyphal cells that form the mycelium. After a certain vegetative growth period, under appropriate conditions, some of the hyphal cells stop normal growth and begin development by forming complex structures called conidiophores that bear multiple chains of conidia (Adams et al. 1988;Park and Yu 2012a).A key and essential step for conidiophore development in Aspergillus is the activation of brlA, which encodes a C 2 H 2 zinc-finger transcription factor (TF) ( Figure 1A) (Adams et al. 1988;Chang and Timberlake 1993). Further genetic and biochemical studies identified the additional key regulators abaA and wetA that function during the middle and late stages of conidiation, respectively ( Figure 1A) (Sewall et al. 1990;Andrianopoulos and Timberlake 1991;Marshall and Timberlake 1991). These three genes have been proposed to define a c...

Functional activation of beta-catenin/T-cell factor (Tcf) signaling has been implicated in human carcinogenesis. We identified the inhibitory effect of various polyphenolic flavonoid compounds against beta-catenin/Tcf signaling in beta-catenin-activated cells. Genistein, kaempferol, isorhamnentin, and baicalein inhibited the transcriptional activity of beta-catenin/Tcf in HEK293 cells transiently transfected with a constitutively active mutant beta-catenin gene. To investigate the inhibitory mechanism, electrophoresis mobility shift assay, immunoprecipitation, and Western blot experiments were performed. The shift assay showed that the binding of Tcf complexes with its specific DNA-binding sites was suppressed by four kinds of flavonoids. Immunoprecipitation analysis also showed that the binding of beta-catenin to Tcf-4 was also disrupted by these flavonoids. Western blot analysis showed a decreased level of beta-catenin in nucleus caused by genistein. Genistein also decreased phosphorylation of Akt and GSK3 beta. Taken together, these results suggest that the polyphenolic flavonoids genistein, kaempferol, isorhamnentin, and baicalein are negative regulators of beta-catenin/Tcf signaling and their inhibitory mechanism is related to the decreased binding of beta-catenin/Tcf complexes to consensus DNA.

We designed a β-CD dimer on silver nanoparticles embedded with silica nanoparticles (Ag@SiO2 NPs) structure to detect polycyclic aromatic hydrocarbons (PAHs). Silica NPs were utilized as a template for embedding silver NPs to create hot spot structures and enhance the surface-enhanced Raman scattering (SERS) signal, and a thioether-bridged dimeric β-CD was immobilized on Ag NPs to capture PAHs. The assembled Ag NPs on silica NPs were confirmed by TEM and the presence of β-CD dimer on Ag@SiO2 was confirmed by UV-vis and attenuated total reflection-Fourier transform infrared spectroscopy. The β-CD dimer@Ag@SiO2 NPs were used as SERS substrate for detecting perylene, a PAH, directly and in a wide linearity range of 10−7 M to 10−2 M with a low detection limit of 10−8 M. Also, the β-CD dimer@Ag@SiO2 NPs exhibited 1000-fold greater sensitivity than Ag@SiO2 NPs in terms of their perylene detection limit. Furthermore, we demonstrated the possibility of detecting various PAH compounds using the β-CD dimer@Ag@SiO2 NPs as a multiplex detection tool. Various PAH compounds with the NPs exhibited their distinct SERS bands by the ratio of each PAHs. This approach of utilizing the assembled structure and the ligands to recognize target has potential for use in sensitive analytical sensors.

BackgroundTemporal hollowing is inevitable after decompressive craniectomy. This complication affects self-perception and quality of life, and various techniques and materials have therefore been used to restore patients’ confidence. Autologous fat grafting in postoperative scar tissue has been considered challenging because of the hostile tissue environment. However, in this study, we demonstrate that autologous fat grafting can be a simple and safe treatment of choice, even for postoperative depressed temporal scar tissue.MethodsAutologous fat grafting was performed in 13 patients from 2011 to 2016. Fat was harvested according to Coleman’s strategy, using a tumescent technique. Patient-reported outcomes were collected preoperatively and at 1-month and 1-year follow-ups. Photographs were taken at each visit.ResultsThe thighs were the donor site in all cases for the first procedure. The median final volume of harvested fat was 29.4 mL (interquartile range [IQR], 24.0–32.8 mL). The median final volume of fat transferred into the temporal area was 4.9 mL on the right side (IQR, 2.5–7.1 mL) and 4.6 mL on the left side (IQR, 3.7–5.9 mL). There were no major complications. The patient-reported outcomes showed significantly improved self-perceptions at 1 month and at 1 year.ConclusionsDespite concerns about the survival of grafted fat in scar tissue, we advise autologous fat grafting for patients with temporal hollowing resulting from a previous craniectomy.

Ethylenediamine-modified β-cyclodextrin (Et-β-CD) was immobilized on aggregated silver nanoparticle (NP)-embedded silica NPs (SiO2@Ag@Et-β-CD NPs) for the effective detection of flavonoids. Silica NPs were used as the template for embedding silver NPs to create hot spots and enhance surface-enhanced Raman scattering (SERS) signals. Et-β-CD was immobilized on Ag NPs to capture flavonoids via host-guest inclusion complex formation, as indicated by enhanced ultraviolet absorption spectra. The resulting SiO2@Ag@Et-β-CD NPs were used as the SERS substrate for detecting flavonoids, such as hesperetin, naringenin, quercetin, and luteolin. In particular, luteolin was detected more strongly in the linear range 10−7 to 10−3 M than various organic molecules, namely ethylene glycol, β-estradiol, isopropyl alcohol, naphthalene, and toluene. In addition, the SERS signal for luteolin captured by the SiO2@Ag@Et-β-CD NPs remained even after repeated washing. These results indicated that the SiO2@Ag@Et-β-CD NPs can be used as a rapid, sensitive, and selective sensor for flavonoids.