Journal: Veterinary sciences

Article Title: Case-Control Study: Endogenous Procalcitonin and Protein Carbonylated Content as a Potential Biomarker of Subclinical Mastitis in Dairy Cows.

doi: 10.3390/vetsci10120670

Figure Lengend Snippet: Figure 1. Results of the Mann–Whitney U test of procalcitonin (PCT) and protein carbonylated content in healthy (H) and subclinical (SCM) groups. The median PCT concentration in the H group was 75.4 pg/mL, whereas in the SCM group, it was 107.3 pg/mL. The PCC median concentration in the H group and SCM group was 0.102 nmol/mL/mg and 0.046 nmol/mL/mg, respectively.

Article Snippet: The concentration of PCT in the plasma samples was determined using a commercial kit for cattle (Bovine Procalcitonin ELISA kit, Cusabio, Houston, TX, USA), as already described in Meucci et al. [53].

Techniques: MANN-WHITNEY, Concentration Assay

Journal: Veterinary sciences

Article Title: Case-Control Study: Endogenous Procalcitonin and Protein Carbonylated Content as a Potential Biomarker of Subclinical Mastitis in Dairy Cows.

doi: 10.3390/vetsci10120670

Figure Lengend Snippet: Figure 2. Receiver operating characteristic (ROC) curves and area under the curve (AUC) for optimal procalcitonin (PCT) cut-off in 130 cows to discriminate the presence or absence of subclinical mastitis.

Article Snippet: The concentration of PCT in the plasma samples was determined using a commercial kit for cattle (Bovine Procalcitonin ELISA kit, Cusabio, Houston, TX, USA), as already described in Meucci et al. [53].

Techniques:

Journal: BMC Veterinary Research

Article Title: Local and systemic effect of transfection-reagent formulated DNA vectors on equine melanoma

doi: 10.1186/s12917-015-0422-9

Figure Lengend Snippet: Treatment substances administered to horses of the three treatment groups on days 1, 22 and 78

Article Snippet: Mologen AG owns a patent for MIDGE-Th1 vectors (PCT/DE02/03798P74).

Techniques: Injection

Journal: BMC Veterinary Research

Article Title: Local and systemic effect of transfection-reagent formulated DNA vectors on equine melanoma

doi: 10.1186/s12917-015-0422-9

Figure Lengend Snippet: Results of RT-qPCR to evaluate in vitro expression of transgenes from MIDGE-Th1 vectors encoding eqIL12, eqIL18, htyr and hgp100

Article Snippet: Mologen AG owns a patent for MIDGE-Th1 vectors (PCT/DE02/03798P74).

Techniques: In Vitro, Expressing

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 ) Schematic outline of the inducible replica-plating work-flow used for inferring repair of DNA double-strand breaks by donor RNA encoded in cgRNAs. ( B ) Dual-expression 2μ plasmid-based designs co-transformed into yeast together with Cas9 alone or with both T7RNAP and Cas9, show that inducible expression of T7RNAP enables efficient repair of Cas9-mediated DSB in the plasmid-encoded ( cis ) artificial intron ( AI ) positioned in the HIS3 open reading frame when expressed in RNase-deficient ( rnh1 rnh201 ) yeast. His + colony forming units (CFUs) out of total colonies are shown. ( C ) Cas9-mediated DSB of HIS3_AI gen in a single-copy genome-encoded ( trans ) his3 AI -disrupted reading frame (Sc71) can be repaired by donor RNA encoded in cgRNAs expressed by inducible T7RNAP in RNase-deficient ( rnh1 rnh201 ) yeast. ( D ) cgRNA expression impacts cgRNA-DNA repair efficiency. A liquid assay was conducted with rnh1 rnh201 strains with the cgRNA construct from pEDJ377 contained in centromeric (CEN/ARS) or 2μ plasmids and expressed from T7 promoter (T7pro) or SNR52 promoter (SNR52pro) as indicated. Genome-integrated HIS3_AI gen was the target, and T7RNAP and Cas9 were inducibly expressed with galactose for 48 hrs prior to plating and His + scoring. Colony-forming units (CFUs) were calculated relative to plating efficiency on non-selective media. ( E ) cgRNA-DNA repair with various donor sizes were investigated as in (D) by symmetric truncations of the cgRNA construct contained in pEDJ377 targeting genome-integrated HIS3_AI gen . For (B-E) frequencies of histidine prototrophic colonies and their error bars are shown as mean ± s.d. from three ( n = 3) biological replicate experiments and significance determined from Student’s t-test, where * p<0.05, ** p<0.005, *** p<0.0005, and N.S. = not significant.

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: Expressing, Plasmid Preparation, Transformation Assay, Construct

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