Journal: The veterinary quarterly

Article Title: Cross-sectional study: can endogenous procalcitonin differentiate between healthy and bovine respiratory disease-affected preweaned dairy calves?

doi: 10.1080/01652176.2024.2434525

Figure Lengend Snippet: Figure 1. Receiver Operating Characteristic (ROC) curves and Area under the curve (AUC) showing the optimal procalcitonin (PCT) cut-off for detecting bovine respiratory disease (BRD) in 226 calves, as diagnosed via thoracic ultrasonography (TUS). BRD is categorized using two common thresholds for lung consolidation: 1 cm and 3 cm. (a) TUS1cm: calves with consolida tions <1 cm (negative) versus those with consolidations ≥1 cm (positive); (b) TUS3cm: calves with consolidations <3 cm (neg ative) versus those with consolidations ≥3 cm (positive).

Article Snippet: The concentration of PCT in plasma samples was measured using a commercial kit designed for cattle (Bovine Procalcitonin ELISA kit, Cusabio, Houston, TX, USA).

Techniques:

Journal: The veterinary quarterly

Article Title: Cross-sectional study: can endogenous procalcitonin differentiate between healthy and bovine respiratory disease-affected preweaned dairy calves?

doi: 10.1080/01652176.2024.2434525

Figure Lengend Snippet: Figure 2. Receiver Operating Characteristic (ROC) curves and Area under the curve (AUC) for the optimal procalcitonin (PCT) cut-off in 226 calves to identify bovine Respiratory dis ease (BRD) diagnosed via the Wisconsin Respiratory Scoring Chart (WRSC). WRSC negative: calves with a WRSC score <5; WRSC positive: calves with a WRSC score ≥5.

Article Snippet: The concentration of PCT in plasma samples was measured using a commercial kit designed for cattle (Bovine Procalcitonin ELISA kit, Cusabio, Houston, TX, USA).

Techniques:

Journal: The veterinary quarterly

Article Title: Cross-sectional study: can endogenous procalcitonin differentiate between healthy and bovine respiratory disease-affected preweaned dairy calves?

doi: 10.1080/01652176.2024.2434525

Figure Lengend Snippet: Figure 3. Receiver Operating Characteristic (ROC) curves and Area under the curve (AUC) for the optimal procalcitonin (PCT) cut-off in 226 calves for the detection of bovine respiratory disease (BRD), diagnosed with a combined approach using the Wisconsin respiratory scoring Chart (WRSC) and thoracic ultrasonography (TUS) with lung consolidation thresholds of 1 cm and 3 cm. (a) WRSC/TUS1cm negative: WRSC < 5 without TUS consolidation; WRSC/TUS1cm positive (upper respiratory tract infec tion): WRSC ≥ 5 without TUS consolidation (b) WRSC/TUS1cm negative: WRSC <5 without TUS consolidation; WRSC/TUS1cm positive (subclinical pneumonia): WRSC < 5 with TUS ≥ 1 cm; (c) WRSC/TUS1cm negative: WRSC < 5 without TUS consolidation; WRSC/TUS1cm positive (clinical pneumonia): WRSC ≥ 5 with TUS ≥ 1 cm; (d) WRSC/TUS3cm negative: WRSC < 5 without TUS consolidation; WRSC/TUS3cm positive (upper respiratory tract infection): WRSC ≥ 5 without TUS consolidation (e)WRSC/TUS3cm negative: WRSC < 5 without TUS consolidation; WRSC/TUS3cm positive (subclinical pneumonia): WRSC < 5 with TUS ≥ 3 cm; (f) WRSC/TUS3cm negative: WRSC < 5 without TUS consolidation; WRSC/TUS3cm positive (clinical pneumonia): WRSC ≥ 5 with TUS ≥ 3 cm.

Article Snippet: The concentration of PCT in plasma samples was measured using a commercial kit designed for cattle (Bovine Procalcitonin ELISA kit, Cusabio, Houston, TX, USA).

Techniques: Infection

Journal: bioRxiv

Article Title: A synthetic RNA-mediated evolution system in yeast

doi: 10.1101/2021.02.27.433199

Figure Lengend Snippet: ( A ) Schematic illustration of experimental set-up, and frequencies of red colonies as a proxy for ade2 knock-out and adenine deficiency. T7 RNA polymerase (T7RNAP) is indispensable for red colony formation in yeast cells expressing Cas9 under the control of the constitutive TEF1 promoter (TEF1pro) and gRNA under the control of the T7 promoter (T7pro), when co-transformed with a linear double-stranded oligo (dsOligo) targeting disruption of ADE2 . Mean frequencies of red colony formation ± s.d. from three ( n = 3) biological replicate experiments. ( B ) Frequencies of red colonies in yeast cells expressing chimeric gRNA (cgRNA) targeting ADE2 ( ADE2 _cgRNA). The cgRNA is based on a 200 nucleotide 5′-primed extension of gRNA homologous to ADE2 with PAM site and four PAM-proximal seed bases deleted (disruption donor; ADE2d). Frequencies from yeast cells expressing either improved Cas9 (iCas9) or Cas9 in the presence or absence of T7RNAP are shown as mean ± s.d. from three ( n = 3) biological replicate experiments.

Article Snippet: CEN.PK2-1C was co-transformed with Cas9 (pEDJ391) or iCas9 (pCT; Addgene #60620) and T7RNAP (pEDJ344) (Sc104 and Sc106, respectively), or an empty vector w/o T7RNAP (p0057) (Sc103 and Sc105, respectively).

Techniques: Knock-Out, Expressing, Control, Transformation Assay, Disruption

Journal: bioRxiv

Article Title: A synthetic RNA-mediated evolution system in yeast

doi: 10.1101/2021.02.27.433199

Figure Lengend Snippet: (A) Plasmid-based galactose-inducible Cas9 and T7RNAP F11L/T613A (epT7RNAP) were expressed with plasmid pEDJ508 (described in the main text). On system induction for 48 hrs in synthetic complete dropout media with galactose, cgRNA_HIS3_stop expressed from pEDJ508 directs Cas9 to the genome-integrated single copy HIS3_23Δ29-XII-5 cassette to induce DNA double-strand break and template DNA repair. The cgRNA may contain the engineered STOP codon sequence (red), or evolved permissive mutations (green) introduced with epT7RNAP. (B) Biological replicates were induced (ON) with galactose dropout media for 48 hrs in 2 ml volumes. 300 μl were plated for each replicate on five plates containing histidine dropout media with glucose to stop evolution (OFF). ( C ) Remaining cells were transferred into liquid histidine dropout media with glucose (OFF) and incubated for 72 hrs to determine growth as indicated by change in OD600 (ΔOD600) during the 72 hrs cultivation in triplicate cultures expressing either pEDJ508 (+cgRNA) or pEDJ400 (-cgRNA). ( D ) Sequencing results from amplicon sequencing of 500 μl of saturated liquid cultures (population level) expressing a repaired HIS3_23Δ29-XII-5 allele (as indicated schematically at the bottom). Corresponding amino acids are shown below HIS3_stop-XII-5 , and TAG (STOP) is boxed and indicated by an asterisk (*). Chromatograms are given for biological replicates #1-3, where blue shading spans the range of mutated bases. Colony scores and OD600 values are presented in Supplementary Table S1.

Article Snippet: CEN.PK2-1C was co-transformed with Cas9 (pEDJ391) or iCas9 (pCT; Addgene #60620) and T7RNAP (pEDJ344) (Sc104 and Sc106, respectively), or an empty vector w/o T7RNAP (p0057) (Sc103 and Sc105, respectively).

Techniques: Plasmid Preparation, Sequencing, Incubation, Expressing, Amplification