Analysis of Alphalactalbumin and Betalactoglobulin from the Rehydration of Bovine Colostrum Powder Using Cloud Point Extraction and Mass Spectrometry

Alphalactalbumin (α-La) and betalactoglobulin (β-Lg) in the rehydration of bovine colostrum powder were successfully separated by cloud point extraction using a nonionic surfactant Triton X-114. The effects of different factors, including the surfactant concentration, sample volume, electrolyte, and pH were discussed. The optimized conditions for cloud point extraction of alphalactalbumin (α-La) and betalactoglobulin (β-Lg) can be concluded that the best surfactant is 1% (w/v) Triton X-114, 200 μL of sample volume, 150 mmol/L NaCl, and 6% (w/v) sucrose. After cloud point extraction, the capillary electrophoresis is used to check the efficiency of the extraction procedure. The results had been effectively confirmed by the characterization with matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS)

Evaluation of Uterosacral Ligament Involvement in Deep Endometriosis by Transvaginal Ultrasonography

This study was designed to conclude the ultrasonic characteristics of uterosacral ligament (USL) lesions involved by endometriosis and evaluated the value of transvaginal sonography (TVS) in diagnosing USL involvement in deep infiltrating endometriosis (DIE). A total of one hundred and eighteen patients with DIE were included in the study and underwent surgery. All these patients were evaluated by transvaginal ultrasound examination by one trained examiner. The gold standard for diagnosis was surgery and histopathology. 85 patients with USL endometriosis were confirmed by surgical pathology. 84 patients were diagnosed USL endometriosis by TVS and 81 of which were confirmed by the gold standard. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy of TVS for diagnosing USL endometriosis were 95.3, 90.9, 96.4, 88.2, and 94.1%, respectively. According to the ultrasound characteristics of USL endometriosis, we summarized four types: Type I. thickened and stiff lesions, Type II. local nodules, Type III. irregular striped lesions, and Type IV. mixed lesions. The conclusion of the study was that TVS was a convenient, accurate and first-line diagnostic technique for USL endometriosis and the USL lesions could be summarized into four types according to the ultrasound morphological changes

Učinkovitost uklanjanja onečišćenja i proizvodnja električne energije pri obradi procjednih voda mikrobnim gorivnim ćelijama s kemijskom katodom i aerobnom biokatodom

The effect of cathode type on contaminant removal efficiency and electricity production in disposing leachate was investigated in a self-assembled microbial fuel cell (MFC). When the landfill leachate was treated with the chemical-cathode MFC (CMFC) and aerobic bio-cathode MFC (ABMFC), the maximum output voltages were 699.0 mV and 459.4 mV, the maximum output powers were 197.7 mW m–3 and 147.6 mW m–3, and the internal resistances were 900 Ω and 700 Ω, respectively. After running the MFCs for 45 days, the COD removal ratios of CMFC and ABMFC were 56.5 % and 64.3 %, the Coulombic efficiencies were 14.3 % and 17.1 %, and the ammonia nitrogen removal ratios were 53.8 % and 58.1 %, respectively. This work is licensed under a Creative Commons Attribution 4.0 International License.Istraživan je utjecaj vrste katode na učinkovitost uklanjanja zagađivala i proizvodnju električne energije pri obradi procjedne vode samoorganizirajućim mikrobnim gorivnim ćelijama (MFC). Kada se procjedna voda s odlagališta obrađivala MFC-om s kemijskom katodom (CMFC) i aerobnom biokatodom (ABMFC), maksimalni izlazni naponi iznosili su 699,0 mV odnosno 459,4 mV, maksimalne izlazne snage 197,7 mW m–3 i 147,6 mW m–3, a unutarnji otpori 900 Ω i 700 Ω. Nakon 45 dana rada gorivnih ćelija, kemijska potrošnja kisika za CMFC i ABMFC smanjena je za 56,5 % i 64,3 %, kulonska učinkovitost bila je 14,3 % i 17,1 %, a uklonjeno je 53,8 % odnosno 58,1 % amonijačnog dušika. Ovo djelo je dano na korištenje pod licencom Creative Commons Imenovanje 4.0 međunarodna

Prime-boost vaccination of mice and rhesus macaques with two novel adenovirus vectored COVID-19 vaccine candidates.

ABSTRACTCOVID-19 vaccines are being developed urgently worldwide. Here, we constructed two adenovirus vectored COVID-19 vaccine candidates of Sad23L-nCoV-S and Ad49L-nCoV-S carrying the full-length gene of SARS-CoV-2 spike protein. The immunogenicity of two vaccines was individually evaluated in mice. Specific immune responses were observed by priming in a dose-dependent manner, and stronger responses were obtained by boosting. Furthermore, five rhesus macaques were primed with 5 × 109 PFU Sad23L-nCoV-S, followed by boosting with 5 × 109 PFU Ad49L-nCoV-S at 4-week interval. Both mice and macaques well tolerated the vaccine inoculations without detectable clinical or pathologic changes. In macaques, prime-boost regimen induced high titers of 103.16 anti-S, 102.75 anti-RBD binding antibody and 102.38 pseudovirus neutralizing antibody (pNAb) at 2 months, while pNAb decreased gradually to 101.45 at 7 months post-priming. Robust T-cell response of IFN-γ (712.6 SFCs/106 cells), IL-2 (334 SFCs/106 cells) and intracellular IFN-γ in CD4+/CD8+ T cell (0.39%/0.55%) to S peptides were detected in vaccinated macaques. It was concluded that prime-boost immunization with Sad23L-nCoV-S and Ad49L-nCoV-S can safely elicit strong immunity in animals in preparation of clinical phase 1/2 trials

FAK Promotes Early Osteoprogenitor Cell Proliferation by Enhancing mTORC1 Signaling

Focal adhesion kinase (FAK) has important functions in bone homeostasis but its role in early osteoprogenitor cells is unknown. We show herein that mice lacking FAK in Dermo1- expressing cells exhibited low bone mass and decreased osteoblast number. Mechanistically, FAK- deficient early osteoprogenitor cells had decreased proliferation and significantly reduced mammalian/mechanistic target of rapamycin complex 1 (mTORC1) signaling, a central regulator of cell growth and proliferation. Furthermore, our data showed that the pharmacological inhibition of FAK kinase- dependent function alone was sufficient to decrease the proliferation and compromise the mineralization of early osteoprogenitor cells. In contrast to the Fak deletion in early osteoprogenitor cells, FAK loss in Col3.6 Cre- targeted osteoblasts did not cause bone loss, and Fak deletion in osteoblasts did not affect proliferation, differentiation, and mTORC1 signaling but increased the level of active proline- rich tyrosine kinase 2 (PYK2), which belongs to the same non- receptor tyrosine kinase family as FAK. Importantly, mTORC1 signaling in bone marrow stromal cells (BMSCs) was reduced if FAK kinase was inhibited at the early osteogenic differentiation stage. In contrast, mTORC1 signaling in BMSCs was not affected if FAK kinase was inhibited at a later osteogenic differentiation stage, in which, however, the concomitant inhibition of both FAK kinase and PYK2 kinase reduced mTORC1 signaling. In summary, our data suggest that FAK promotes early osteoprogenitor cell proliferation by enhancing mTORC1 signaling via its kinase- dependent function and the loss of FAK in osteoblasts can be compensated by the upregulated active PYK2. © 2020 American Society for Bone and Mineral Research.Schematic model of the differential roles of FAK in the cells of osteoblast lineage. The model depicts the mechanisms of FAK action at three distinct stages of osteoblast lineage in which the roles of FAK have been addressed by genetic and pharmacological approaches as well as the respective Cre transgenes used to target Fak, including Dermo1- Cre (this study), Osterix- Cre,(10) Col3.6- Cre (this study), and Col2.3- Cre.(9) Red - indicates that the loss of FAK in osteoblasts can be compensated by the upregulated active PYK2.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/162813/3/jbmr4029-sup-0001-Supinfo.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/162813/2/jbmr4029_am.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/162813/1/jbmr4029.pd

Application of Industrial Standard Methods for Detection of Horse- and Donkey-Derived Ingredients for Detecting Mule Meat

Both horse- and donkey-derived ingredients have been detected in mule meat by real-time polymerase chain reaction (PCR) as described in China’s industry standards for detection of horse (SN/T 3730.5-2013) and donkey (SN/T 3730.4-2013) ingredients in food and feed, respectively. This contradicts the theory of strict maternal inheritance of mitochondrial DNA (mtDNA). Therefore, in this study, 3 horse meat samples, 3 donkey meat samples and 3 mule meat samples were detected by mitochondrial gene and nuclear gene sequencing based on PCR and the China’s industry standard methods for horse and donkey ingredients, respectively, and the results of the SN/T 3730.4-2013 method for mule meat were analyzed. According to the results of mitochondrial gene and nuclear gene sequencing, all 3 mule meat samples were derived from mules. Both donkey and horse ingredients were detected in the 3 mule meat samples by the SN/T 3730.4-2013 and SN/T 3730.5-2013 methods. The cycle threshold (Ct) of the SN/T 3730.5-2013 method for horse ingredient was in the range of ≤ 20.00, and that of the SN/T 3730.4-2013 method for donkey ingredient were in the range of 25.00-35.00. The sequencing results of PCR products using the primers described in the SN/T 3730.4-2013 method showed that the 3 mule meat samples had no homology with horse or donkey meat. This might be because the SN/T 3730.4-2013 target sequence appeared in the form of nuclear mitochondrial DNA segments in low repeat numbers in the mule nuclear genome, and some base insertions and deletions occurred. The possibility that mule ingredient may be present should be considered when the Ct value of the SN/T 3730.4-2013 is ≤ 20.00, while the Ct value of the SN/T 3730.5-2013 is in the range of 25.00-35.00 for horse and donkey ingredients in known samples of single animal-derived ingredients, respectively