Molekularna karakterizacija gena za mitohondrijsku 16S rRNA goveda, bivola i jaka.

A combination of polymerase chain reaction (PCR) with restriction fragment length polymorphism (RFLP) and nucleotide sequencing is the most preferred and efficient method for characterization of different species, in terms of detection power and applicability to large scale screening. The present study was carried out with the aim of developing the molecular fingerprint of the mitochondrial 16S rRNA gene of Cattle, Buffalo and Yak. Blood samples were collected randomly from ten different animals of each species for mitochondrial DNA extraction. The extracted DNA was used for the amplification of the 16S rRNA gene using universal primers. The size of the amplified products was 600bp. RFLP studies were carried out by digesting the amplicons using restriction enzymes viz. AluI, HinfI and HaeIII. The resulting RFLP pattern could easily identify and differentiate each of the species. Sequencing of the amplicons in all three species was carried out to confirm the variations at nucleotide level. Sequence analysis of the 16S rRNA gene using MEGA4 software and also PCRRFLP revealed that the 16S rRNA gene can be used as a good candidate for a molecular marker.Kombinacija lančane reakcije polimerazom (PCR) s polimorfizmom duljine restrikcijskih fragmenata (RFLP) i utvrđivanjem slijeda nukleotida (sekvenciranje) najpoželjniji je i učinkovit postupak za otkrivanje širokog raspona varijacija pri karakterizaciji različitih vrsta. Cilj ovog istraživanja bio je razviti molekularni otisak za gen koji određuje mitohondrijske 16S rRNA goveda, bivola i jaka. Za izdvajanje mitohondrijske DNA nasumično je prikupljeno po 10 uzoraka krvi različitih životinja unutar svake vrste. Izdvojena DNA iskorištena je za umnažanje 16S rRNA gena uz uporabu univerzalnih početnica. Veličina umnoženih proizvoda iznosila je 600 bp. RFLP analize provedene su digestijom amplikona pomoću restrikcijskih enzima viz. AluI, HinfI i HaeIII. Dobiveni RFLP uzorak mogao je lako prepoznati i razlikovati svaku od vrsta. Sekvenciranje amplikona u sve tri vrste provedeno je s ciljem da se potvrde varijacija na razini nukleotida. Analiza sekvencija pomoću računalnog programa MEGA4 i metoda PCR-RFLP pokazala je da se gen 16S rRNA može upotrijebiti kao dobar kandidat za molekularni biljeg

Effect of High-Flow Nasal Cannula Therapy vs Continuous Positive Airway Pressure Following Extubation on Liberation From Respiratory Support in Critically Ill Children

IMPORTANCE: The optimal first-line mode of noninvasive respiratory support following extubation of critically ill children is not known. OBJECTIVE: To evaluate the noninferiority of high-flow nasal cannula (HFNC) therapy as the first-line mode of noninvasive respiratory support following extubation, compared with continuous positive airway pressure (CPAP), on time to liberation from respiratory support. DESIGN, SETTING, AND PARTICIPANTS: This was a pragmatic, multicenter, randomized, noninferiority trial conducted at 22 pediatric intensive care units in the United Kingdom. Six hundred children aged 0 to 15 years clinically assessed to require noninvasive respiratory support within 72 hours of extubation were recruited between August 8, 2019, and May 18, 2020, with last follow-up completed on November 22, 2020. INTERVENTIONS: Patients were randomized 1:1 to start either HFNC at a flow rate based on patient weight (n = 299) or CPAP of 7 to 8 cm H2O (n = 301). MAIN OUTCOMES AND MEASURES: The primary outcome was time from randomization to liberation from respiratory support, defined as the start of a 48-hour period during which the child was free from all forms of respiratory support (invasive or noninvasive), assessed against a noninferiority margin of an adjusted hazard ratio (HR) of 0.75. There were 6 secondary outcomes, including mortality at day 180 and reintubation within 48 hours. RESULTS: Of the 600 children who were randomized, 553 children (HFNC, 281; CPAP, 272) were included in the primary analysis (median age, 3 months; 241 girls [44%]). HFNC failed to meet noninferiority, with a median time to liberation of 50.5 hours (95% CI, 43.0-67.9) vs 42.9 hours (95% CI, 30.5-48.2) for CPAP (adjusted HR, 0.83; 1-sided 97.5% CI, 0.70-∞). Similar results were seen across prespecified subgroups. Of the 6 prespecified secondary outcomes, 5 showed no significant difference, including the rate of reintubation within 48 hours (13.3% for HFNC vs 11.5 % for CPAP). Mortality at day 180 was significantly higher for HFNC (5.6% vs 2.4% for CPAP; adjusted odds ratio, 3.07 [95% CI, 1.1-8.8]). The most common adverse events were abdominal distension (HFNC: 8/281 [2.8%] vs CPAP: 7/272 [2.6%]) and nasal/facial trauma (HFNC: 14/281 [5.0%] vs CPAP: 15/272 [5.5%]). CONCLUSIONS AND RELEVANCE: Among critically ill children requiring noninvasive respiratory support following extubation, HFNC compared with CPAP following extubation failed to meet the criterion for noninferiority for time to liberation from respiratory support. TRIAL REGISTRATION: isrctn.org Identifier: ISRCTN60048867