CDK7 Regulates the Mitochondrial Localization of a Tail-Anchored Proapoptotic Protein, Hid

SummaryThe mitochondrial outer membrane is a major site of apoptosis regulation across phyla. Human and C. elegans Bcl-2 family proteins and Drosophila Hid require the C-terminal tail-anchored (TA) sequence in order to insert into the mitochondrial membrane, but it remains unclear whether cytosolic proteins actively regulate the mitochondrial localization of these proteins. Here, we report that the cdk7 complex regulates the mitochondrial localization of Hid and its ability to induce apoptosis. We identified cdk7 through an in vivo RNAi screen of genes required for cell death. Although CDK7 is best known for its role in transcription and cell-cycle progression, a hypomorphic cdk7 mutant suppressed apoptosis without impairing these other known functions. In this cdk7 mutant background, Hid failed to localize to the mitochondria and failed to bind to recombinant inhibitors of apoptosis (IAPs). These findings indicate that apoptosis is promoted by a newly identified function of CDK7, which couples the mitochondrial localization and IAP binding of Hid

Isolation and characterization of the outer membrane of Escherichia coli with autodisplayed Z-domains

Abstract“Autodisplay technology” is an expression technique used to display the various recombinant proteins on the outer membrane (OM) of Escherichia coli. The resulting autodisplayed Z-domain has been used to improve the sensitivity of immunoassays. In this work, a facile isolation method of the OM fraction of E. coli with autodisplayed Z-domains was presented using (1) an enzyme reaction for the hydrolysis of the peptidoglycan layer and (2) short centrifugation steps. The purity of the isolated OM fraction was analyzed. For the estimation of contamination with bacterial proteins from other parts of E. coli, Western blots of marker proteins for the OM (OmpA), periplasm (β-lactamase), inner membrane (SecA), and cytoplasm (β-galactosidase) were performed. Additionally, assays of marker components or enzymes from each part of E. coli were carried out including the OM (KDO), inner membrane (NADH oxidase), periplasm (β-lactamase), and cytoplasm (β-galactosidase). The yield of OM isolation using this new method was determined to be 80% of the total OM amount, with less than 1% being contaminants from other parts of E. coli

Bis(2-amino-5-methyl-1,3,4-thia­diazole-κN 3)dichloridocobalt(II)

In the monomeric title complex, [CoCl2(C3H5N3S)2], the CoII atom is tetra­coordinated by two chloride anions and two N atoms from two monodentate 2-amino-5-methyl-1,3,4-thia­diazole ligands, giving a slightly distorted tetra­hedral stereochemistry [bond angle range about Co = 105.16 (12)–112.50 (10)°]. In the complex, the dihedral angle between the 1,3,4-thia­diazole planes in the two ligands is 72.8 (1)°. There are two intra­molecular N—H⋯Cl inter­actions in the complex unit, while in the crystal, inter­molecular N—H⋯N and N—H⋯Cl hydrogen bonds link these units into a two-dimensional layered structure parallel to (011)

Pinoresinol diglucoside exhibits protective effect on dexamethasone-induced osteoporosis in rats

Purpose: To investigate the effect of pinoresinol diglucoside (PDG) on dexamethasone-induced osteoporosis in rats.Methods: Sixty Wistar rats were randomly and equally divided into normal, control, alendronate and PDG (10, 20 or 40 mg/kg) groups. Bone tissue parameters, including length, transverse diameter, weight, bone mineral content (BMC) and bone mineral density (BMD), were determined using vernier caliper, electronic balance and single photon bone mineral density meter. Serum biochemical indices, including Ca2+, inorganic phosphorus (IP), IL-6, TNF-α and alkaline phosphatase (ALP), were determined using colorimetry and enzyme-linked immunosorbent assay (ELISA). Osteoprotegerin (OPG) and receptor activator of nuclear factor-κB ligand (RANKL) proteins were detected by Western blot.Results: PDG (10, 20 or 40 mg/kg) increased significantly (p < 0.05 or 0.01) transverse diameter (3.64 – 3.79 vs. 3.31 mm), weight (0.73 – 0.78 vs. 0.67 g), BMC (0.16 – 0.23 vs. 0.12 g/cm), BMD (0.27 – 0.35 vs. 0.22 g/cm2) of right femur, serum Ca2+ level (2.16 – 2.39 vs. 1.94 mmol/L), and OPG level of left femur, compared with those in the control group. PDG (10, 20 or 40 mg/kg) reduced significantly (p < 0.05 or 0.01) serum IP (1.34 – 1.14 vs. 1.76 mmol/L), IL-6 (103.25 – 95.38 vs. 108.74 ng/L), TNF-α (87.46 – 82.05 vs. 92.38 ng/L), ALP (334.79 – 276.32 vs. 486.45 U/L) levels or activities, and RANKL level of left femur, compared with those in the control group.Conclusion: PDG exhibits a protective effect on dexamethasone-induced osteoporosis by increasing bone mass and regulating bone metabolism. Thus, PDG may be a candidate drug for treating osteoporosis.Keywords: Pinoresinol diglucoside, Osteoporosis, Bone mass, Bone metabolism, Dexamethasone, Osteoprotegeri

Factors influencing pregnancy stress in pregnant women in Korea: a cross-sectional study

Purpose The purpose of this study was to investigate the association between maternal knowledge and social support on pregnancy stress among pregnant women in Korea. Methods The participants in this study were 148 pregnant women in Korea, recruited from online communities on pregnancy and/or childbirth, from June 2019 to April 2020. The collected data were analyzed using the independent t-test, one-way analysis of variance, Pearson correlation coefficient, and multiple regression. Results Participants were at average 18.25±8.28 weeks gestation, 56% were in the second trimester, 31% had one or more health issues in the current pregnancy (e.g., hyperemesis gravidarum), and 76% were first-time mothers. Participants had moderate levels of pregnancy stress (mean, 23.09±7.11 points out of 48) and maternal knowledge (mean, 14.42±4.67 points out of 21), whereas social support was somewhat high (mean 45.88±7.81 points out of 60). Pregnancy stress was weakly negatively correlated with social support (r=–.37, p<.001). Main source of pregnancy information (β=–.21, p=.011), marital satisfaction (β=–.18, p=.036), and social support (β=–.19, p=.038) were identified as significant factors affecting pregnancy stress, and these variables had an explanatory power of 22.7% for pregnancy stress. Conclusion Based on these findings, nurses should assess pregnancy-related stress during pregnancy and consider main source of pregnancy information and marital satisfaction when providing education or counseling. Moreover, strategies to reduce pregnancy stress through social support are needed to improve the quality of life for pregnant women

Flightless-I Controls Fat Storage in Drosophila

Triglyceride homeostasis is a key process of normal development and is essential for the maintenance of energy metabolism. Dysregulation of this process leads to metabolic disorders such as obesity and hyperlipidemia. Here, we report a novel function of the Drosophila flightless-I (fliI) gene in lipid metabolism. Drosophila fliI mutants were resistant to starvation and showed increased levels of triglycerides in the fat body and intestine, whereas fliI overexpression decreased triglyceride levels. These flies suffered from metabolic stress indicated by increased levels of trehalose in hemolymph and enhanced phosphorylation of eukaryotic initiation factor 2 alpha (eIF2??). Moreover, upregulation of triglycerides via a knockdown of fliI was reversed by a knockdown of desat1 in the fat body of flies. These results indicate that fliI suppresses the expression of desat1, thereby inhibiting the development of obesity; fliI may, thus, serve as a novel therapeutic target in obesity and metabolic diseases

Comparative analysis of FBS containing media and serum free chemically defined media, CellCor for adipose derived stem cells production

Background: As a result of the aging society, the average OECD life expectancy has grown to about 80 years, yet the average health life still remains at only 65 years, leaving more than 15 years of life in an uncertain health state. Regenerative medicine is a new concept of medicine that combines cells and biomaterials to restore the functions of aged or damaged tissues or organs. It is also a good treatment for chronic diseases and incurable diseases, receiving attention as a new paradigm for treating diseases. Problems: As the market for regenerative medicine grows, mass production of consistent quality cells is required. Media is the most important thing in mass production of consistent quality cells. However, the fetal bovine serum (FBS) containing media that is currently wide used has many problems, such as unidentified viral infection, immunogenicity, lot variations, unstable supply, and ethical issues. To solve these problems and make rapid progress in regenerative medicine, a high-performance serum free chemically defined media (CDM) is needed. Solution: CellCor is a serum free CDM that provides excellent performance, safety, economy and consistency in stem cell production. CellCor allows higher-speed cell production rate than current FBS containing culture media (Figure 1). Compared to the FBS containing media, CellCor is able to maintain stem cell markers, higher population homogeneity, genetic stability, and excellent differentiation potency even at later passage. Please click Additional Files below to see the full abstract

Genetic Association of NPY Gene Polymorphisms with Dampness-Phlegm Pattern in Korean Stroke Patients

Neuropeptide Y (NPY), which is widely expressed in both the central and peripheral nervous systems, has an important role in a variety of biological fields. In this study, we analyzed the distribution of NPY polymorphisms in dampness-phlegm pattern and non-dampness-phlegm pattern in elderly Korean subjects with cerebral infarction (CI). A total of 1.097 subjects (498 normal subjects and 599 CI patients, including 198 with dampness-phlegm pattern and 401 with non-dampness-phlegm pattern) participated in this study. Genotyping for five SNPs (G-1484A, C-1471T, C-399T, A1201G, and C5325T) was conducted by primer extension. The results were statistically analyzed for genetic association of NPY-polymorphisms with normal versus dampness-phlegm pattern or non-dampness-phlegm pattern subjects. Among the five SNPs tested, the T allele of C-399T has a negative association with the dampness-phlegm pattern and is marked by a decrease in serum cholesterol levels. Furthermore, serum cholesterol levels were significantly higher in dampness-phlegm pattern patients than in non-dampness-phlegm pattern patients.In this study, for the first time, the association of NPY polymorphisms with pattern identification (PI) of traditional Korean medicine (TKM) was analyzed in a large CI patient population