Joanna Kochan, Wojciech Niżański, Nei Moreira, Zalmir Silvino Cubas, Agnieszka Nowak, Sylwia Prochowska, Agnieszka Partyka, Wiesława Młodawska and Józef Skotnicki
disciplines of science, starting from the biology of the species, through research into the population and habitat, assisted reproductive technologies, establishment of gene banks, developing bioinformatic systems, and ending with biodiversity and endangered species management.
In the last few years knowledge of felid reproduction has expanded considerably thanks to comparative studies utilising the domestic cat as a research model for endangered wild cats. Basic reproductive techniques used both in domestic cat breeding and in rescuing wild felid populations that are
traditional PCR, nested PCR, real-time quantitative PCR, nucleic acid sequence-based amplification (NASBA), loop-mediated isothermal amplification (LAMP) technology, and RNA simultaneous amplification and testing (SAT). On the basis of the basic principles of these experimental methods, domestic and foreign scholars have developed a composite technology that can simultaneously detect multiple pathogens. The target genes frequently amplified in the experiment include P1 protein coding gene, 16S rRNA, card gene, ATPase gene, etc. The types of clinical specimens available are
application and development of omics technology, a microecological system that interacts with hosts has been found in the lungs, and the microbiome plays an important role in maintaining the health of the respiratory tract.
Respiratory infections and disease progression are associated with the regulation of microbial immunity in colonization. In lower respiratory tract infections in infants at early stages, changing the colonization structure of the original respiratory tract increases the chances of suffering from allergic reactions and repeated wheezing [ 9 , 10 ]. In
May Phonvisay, Jai-Wei Lee, Jhong-Jie Liou, Hsian-Yu Wang and Chun-Yen Chu
University of Science and Technology, Taiwan.
RA-specific antibody responses . The RA-specific antibody levels of immunised ducks were examined using indirect ELISA as previously described ( 4 ). RA cell lysate was prepared by coating it onto 96-well plates at 0.5 μg/well. The scotopic to protopic (S/P) ratio was calculated as (test OD value − negative OD value) / (positive OD value − negative OD value).
Cytokine expression analysis . The mRNA levels of cytokine genes were analysed as previously described ( 4 ). Briefly, PBMCs from vaccinated ducks were collected and
Yu-Xi Song, Pan Hu, Yun-Long Bai, Chang Zhao, Cheng Xia and Chuang Xu
temperature to dryness. The dry extract was then redissolved in 100 μL of acetonitrile:H 2 O (1:1, v/v), vortexed for 30 s, sonicated for 10 min in an ice bath, and centrifuged at 12,000 × g for 15 min at 4°C to remove insoluble debris. From each treated sample, 5 μL of the supernatant was mixed as a quality control (QC) sample and 70 μL was aliquoted for LC–MS detection ( 12 ).
Detection analysis . Detection was performed using an Agilent 1290 Infinity II LC system (Agilent Technologies, USA) coupled to a TripleTOF 6600 System (Q-TOF; AB Sciex, USA).
Esraa A. Elshafiee, Sara M. Nader, Sohad M. Dorgham and Dalia A. Hamza
gel to determine the size of the product.
Sequence analysis of toxA gene. After selecting one isolate each from animal, water, and human samples that showed resistance to carbepenem genes, the amplified toxA fragments were purified using the QIAquick gel extraction kit (Qiagen, Germany) according to the manufacturer’s instructions and sequenced at Promega Lab Technology (Germany) by using the forward and reverse primers of toxA listed in Table 1 . The sequences of the toxA gene were deposited in the National Center for Biotechnology Information (NCBI