Body growth and blood metabolite concentrations in Boran and Boran x Friesian bulls grazed on natural pastures: Effect of dry season dietary supplementation

Twenty-seven Boran and 37 Boran x Friesian bulls, weaned at six months of age, were allocated to either receive supplementary diet (16 percent crude protein and 10.8 MJ/kg DM energy) during the dry season or to serve as unsupplemented controls until 21 months of age. Body weight measurements were taken monthly and two sets of blood samples were collected from all bulls during each of the two dry seasons and one intervening wet season during the 15-month study period. Plasma samples were used for determination of plasma total protein, albumin, globulin, blood urea nitrogen (BUN) and glucose concentrations. Over the study period, overall body weight gain was positive for all treatment groups, but was higher in supplemented than in control bulls and in Boran x Friesian than in Boran bulls. Within genotype, supplementary feeding increased overall body growth by 2.2- and 1.6-fold in Boran and Boran x Friesian bulls, respectively. Within genotype, supplemented bulls had higher total protein and albumin levels, while plasma globulin, BUN and glucose were not affected. On average, Boran bulls had lower concentrations of total protein and globulin, but higher albumin and BUN levels than the Boran x Friesian bulls. Blood glucose concentrations were not influenced by dietary supplementation, did not differ between genotypes and showed no consistent trends over time in the two genotypes. Body weight was positively correlated with plasma total protein (0.54) and globulin (0.49) concentrations, but negatively related with albumin/globulin ration (-0.48) and blood urea nitrogen (-0.57). It was concluded that the body growth of young bulls grazing natural pastures during the wet season was adequate and dry season dietary supplementation improved growth in both Boran and Boran x Friesian bulls. Determination of blood metabolites considered in this study did not assist in assessing the nutritional status of young Boran and Boran x Friesian bulls using the types of feeds and at the level of supplementation provided

<i>Il17ra</i> expression in the gut of control and <i>S</i>.Tm^*-infected C57BL/6 mice.

<p>(A) <i>Il17a</i>, <i>Il17f</i> and <i>Il17ra</i> expression in the intestinal tissues of naïve mice (n = 3 per group). Relative levels of mRNA in 20 ng total RNA were normalized to <i>house keeping genes</i>. ΔCt values do not strictly correlate to the difference in gene expression between various genes. However, ΔCt values <19 represent detectable levels of expression, demonstrating thereby that all three genes are expressed in the various tissue segments. (B) Induction of <i>Il17ra</i> expression in <i>Salmonella</i>-infected animals. <i>Salmonella</i>-infected animals (n = 3; 12 h p.i. with <i>S</i>.Tm*; open circles) were from the experiment shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0013804#pone-0013804-g002" target="_blank">Fig. 2</a>. Non-infected, stereptomycin pretreated C57BL/6 mice (n = 3; black circles) served as a contol. Samples of gut tissues (A, as indicated; B, cecum tissue) were taken for RNA isolation and mRNA levels were analyzed by real time PCR. <i>S</i>.Tm^* infected C57BL/6 mice showed a significant up-regulation of <i>Il17ra</i> in comparision to non-infected C57BL/6 mice (p<0.05; t-test unpaired, two-tailed). Sm: Streptomycin. Line: median.</p

Effect of IL-17A and/or F immunization on the <i>S.</i>Typhimurium infection.

<p>C57BL/6 mice were vaccinated three times (50 µg each, s.c.) with IL-17A-VLP, IL-17F-VLP, either reagents or unconjugated control VLPs. A–C) αIL-17A and αIL-17F titers were analyzed two weeks after the last immunization by ELISA and compared to unconjugated-VLP immunized controls (B,C; OD450 nm values +/− SEM). D,E) Neutralization was tested by ELISA. D) ELISA plates were coated with 1 µg/ml IL-17RA and binding of 10 ng/ml biotinylated IL-17A to IL-17RA was tested in the presence of serum of mice immunized with IL-17A-VLPs, IL-17F-VLPs or VLPs alone (OD450 nm values +/− SEM). E) ELISA plates were coated with 1 µg/ml IL-17RC and binding of 200 ng/ml biotinylated IL-17F to IL-17RC was tested in the presence of serum of mice immunized with IL-17A-VLPs, IL-17F-VLPs or VLPs alone; OD450 nm values +/− SEM. F–J) Subsequent analysis in <i>S</i>. Typhimurium challenge infections. Animals were pretreated with streptomycin and infected for 2 days with wt <i>S</i>. Typhimurium. PBS immunization and PBS treatment served as control. We analyzed colonization levels in the gut lumen (F), the mLN (G), the liver (H) and the spleen (I), as well as the degree of inflammation of the cecum mucosa (J). Comparison with the control VLPs immunized mice did not reveal any significant differences in any of the parameters of the infection, analyzed (J: Mann-Whitney U test was not significant as well as ANOVA with Bonferroni (p = 0.0841)). Stippled line: minimal detectable value.</p

<i>S</i>.Typhimurium infection of <i>Il17ra^−/−</i> mice.

<p><i>Il17ra^−/−</i> mice, <i>Il17ra^+/−</i> littermates and C57BL/6 control mice were pretreated with streptomycin and infected for 12 hours with <i>S</i>.Tm* or isogenic derivatives <i>S</i>.Tm^SipA, <i>S</i>.Tm^SopE, <i>S</i>.Tm^avir (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0013804#s2" target="_blank">Material and Methods</a>). We analyzed colonization levels in the gut lumen (A) and the mLN (B), as well as the degree of inflammation of the cecum mucosa (C). n.s.: No significant difference between <i>Il17ra^−/−</i> mice and <i>Il17ra^+/−</i> littermates. Strippled line: minimal detectable value.</p

PTN expression level and its stimulation effect on the proliferation of B16-F10 melanoma cells.

<p>(A). Western-blot analysis of the expression level of PTN in B16-F10 melanoma cells and HEK293 cells. β-actin was detected as a load control. (B). Effects of recombinant PTN on B16-F10 melanoma cell proliferation. Each point represented the mean ± standard deviation of 3 replicates. PTN stimulated B16-F10 melanoma cells proliferation before the cells reached full confluency at a concentration of 100 ng/ml (P < 0.01 compared with control without adding the extracellular PTN). (C). Cells numbers of B16-F10 melanoma 24 hours after different treatments were counted and compared with the control group. Each bar represented the mean ± standard deviation of 3 replicates. Asterisks indicate statistically significant differences between samples (** p<0.01).</p

Monitoring the liver toxicity of mice in different treatment groups.

<p>Mice liver enzyme ALT (A) and AST (B) levels were detected at the end of study. There was no significant difference among the groups. The enzyme levels were in the same range as the group treated with normal saline. Data were shown as the mean ± SD. (C) Western blotting analysis of the PTN expression level in the lungs of mice in different treatment groups.</p

All three different systemic treatments by PTN siRNAc inhibited the tumor growth.

<p>(A). The microscopic image of the 60% glucose microbubble with 25x magnification. (B). The microscopic image of the 60% glucose microbubble filtered through the grade 595 1/2 filter paper with 25x magnification. Scale bar is 50 μm for both images. (C-E). Therapeutic effect of the systemic treatment of PTN siRNAc by using jetPEI, TfPEI and microbubble-jetPEI. (C). Lung of the mice treated with jetPEI-random sequence (Top) and jetPEI-siRNAc (Bottom). (D). Lung of the mice treated with TfPEI-random sequence (Top) and TfPEI-siRNAc (Bottom). (E). Lung of the mice treated with microbubble-jetPEI random sequence (Top) and microbubble-jetPEI siRNAc (Bottom). (F-G). The weighted sum of the metastatic colony numbers was plotted in bar chart as mean± standard deviation of 5 mice in each group. (G). The comparison of the siRNAc treatment group and the NC treatment group with the same delivery method. (F). The comparison between different siRNAc delivery methods. Asterisks indicate statistically significant differences between samples (** p<0.01 and * p<0.05) (H-K). Melanoma metastasis in liver and gastrointestinal tract. (H). The mice treated with random sequence showed a tumor metastasis in the gastrointestinal tract and liver. (I-K). The mice treated with siRNAc by systemic using of all three <i>in-vivo</i> transfection agent: (I). jetPEI-siRNAc, (J). TfPEI-siRNAc, (K). microbubble-jetPEI siRNAc revealed no tumor metastasis in the gastrointestinal tract and liver.</p

The silence efficiency of PTN siRNA measured by quantitative RT-PCR and western blotting.

<p><b>The PTN expression level showed a direct correlation with the proliferation rate of B16-F10 melanoma cells.</b> (A). Quantitative RT-PCR analysis of the knock-down efficiency of siRNA 1, siRNA 2 and the mixture of them at the same dose. GAPDH was used as a control. (B). Western blotting analysis of the knock-down effect of siRNA 1, siRNA 2 and the mixture of them at the same dose. β-actin was used as a control. The intensity of the control sample without transfection of any siRNA was set as 100%. Asterisks indicate statistically significant differences between samples (** p<0.01 and ns for no significant difference). (C). Silence of the PTN by siRNA reduced the proliferation rate of B16-F10 melanoma cells in a manner directly related the reduced level of PTN. Each point represents the mean ± standard deviation of 3 replicates.</p

Baseline data stratified by virus type and variant.

Baseline data stratified by virus type and variant.</p

Model summary: Prediction of mortality.

BackgroundWith the emergence of new subvariants, the disease severity of Severe Acute Respiratory Syndrome Coronavirus-2 has attenuated. This study aimed to compare the disease severity in patients hospitalized with omicron variant infection to those with influenza infection.MethodsWe compared data from the multicenter observational, prospective, epidemiological “CORONA Germany” (Clinical Outcome and Risk in hospitalized COVID-19 patients) study on patients infected with Severe Acute Respiratory Syndrome Coronavirus-2 to retrospective data on influenza infection cases from November 2016 to August 2022. Severe Acute Respiratory Syndrome Coronavirus-2 cases were classified as wild-type/delta variant before January 2022, or omicron variant from January 2022 onward. The primary outcome was in-hospital mortality, adjusted for age, gender, and comorbidities.ResultsThe study included 35,806 patients from 53 hospitals in Germany, including 4,916 patients (13.7%) with influenza infection, 16,654 patients (46.5%) with wild-type/delta variant infection, and 14,236 patients (39.8%) with omicron variant infection. In-hospital mortality was highest in patients with wild-type/delta variant infection (16.8%), followed by patients with omicron variant infection (8.4%) and patients with influenza infection (4.7%). In the adjusted analysis, higher age was the strongest predictor for in-hospital mortality (age 80 years vs. age 50 years: OR 4.25, 95% CI 3.10–5.83). Both, patients with wild-type/delta variant infection (OR 3.54, 95% CI 3.02–4.15) and patients with omicron variant infection (OR 1.56, 95% CI 1.32–1.84) had a higher risk for in-hospital mortality than patients with influenza infection.ConclusionAfter adjusting for age, gender and comorbidities, patients with wild-type/delta variant infection had the highest risk for in-hospital mortality compared to patients with influenza infection. Even for patients with omicron variant infection, the adjusted risk for in-hospital mortality was higher than for patients with influenza infection. The adjusted risk for in-hospital mortality showed a strong age dependency across all virus types and variants.Trial registration numberNCT04659187.</div