Introducing forage in the young calf diet during the milk-feeding period stimulates rumen development. It was hypothesized that performance in dairy calves would depend on forage provision and starter physical form such that the textured starter (TS) feed with corn silage (CS) supplementation would benefit calf performance. This study evaluates the effects of the physical form of starter diets and CS supplementation on performance, rumen fermentation characteristics, and structural growth of dairy calves. Forty-eight 3-d-old Holstein dairy calves with a mean starting BW of 42.1 kg (SD 2.4) were used in a 2 × 2 factorial arrangement with the factors dietary CS level (0 or 15% on DM basis) and physical form of starter (mashed vs. textured). Individually housed calves were randomly assigned ( = 12 calves per treatment: 6 males and 6 females) to 4 treatments: 1) a mashed starter (MS) feed with no CS (MS-NCS), 2) a MS feed with CS (MS-CS), 3) a TS feed with no CS (TS-NCS), and 4) a TS feed with CS (TS-CS). The calves had ad libitum access to water and starter throughout the study. All calves were weaned on d 56 of age and remained in the study until d 66. The interaction of the physical form of the starter and CS provision was significant ( < 0.01) for the starter intake, with the greatest intake for TS-CS treatment during the preweaning and overall periods. Regardless of the physical form of starter, starter intake, ADG, weaning BW, final BW, ruminal pH, the molar proportion of acetate, and the acetate-to-propionate ratio were greater ( < 0.01) for CS-supplemented calves compared with unsupplemented calves. No interaction ( > 0.05) was detected between the physical form of starter and CS provision with respect to the rumen fermentation parameters and body measurements. Total rumen VFA concentration and the molar proportion of propionate were greater ( < 0.01) in calves fed TS compared with MS-fed calves. In conclusion, independent of the physical form of starter, inclusion of 15% CS in starter diets improves the performance of dairy calves.

A meta-analysis of the potential effect of forage provision on growth performance and rumen fermentation of dairy calves was conducted using published data from the literature (1998-2016). Meta-regression was used to evaluate the effects of different forage levels, forage sources, forage offering methods, physical forms of starter, and grain sources on the heterogeneity of the results. We considered 27 studies that reported the effects of forage provision to dairy calves. Estimated effect sizes of forage were calculated on starter feed intake, average daily gain (ADG), feed efficiency (FE), body weight (BW), and rumen fermentation parameters. Intake of starter feed, ADG, BW, ruminal pH, and rumen molar proportion of acetate increased when supplementing forage but FE decreased. Heterogeneity (the amount of variation among studies) was significant for intake of starter feed, ADG, FE, final BW, and rumen fermentation parameters. Improving overall starter feed intake was greater in calves offered alfalfa hay compared with those offered other types of forages. During the milk feeding and overall periods, improving ADG was greater for calves fed a high level of forage (>10% in dry matter) compared with those fed a low level of forage (≤10% in dry matter) diets. The advantages reported in weight gain at a high level of forage could be due to increased gut fill. Improving overall ADG was lower for calves offered forages with textured starter feed compared with ground starter feed. The meta-regression analysis revealed that changes associated with forage provision affect FE differently for various forage sources and forage offering methods during the milk-feeding period. Forage sources also modulated the effect of feeding forage on ruminal pH during the milk-feeding period. In conclusion, forage has the potential to affect starter feed intake and performance of dairy calves, but its effects depend on source, level, and method of forage feeding and physical form of starter feed independently of grain sources included in the starter feed.

Termination of wound-healing process requires a fine balance between connective tissue deposition and its hydrolysis. Previously, we have demonstrated that keratinocyte-releasable stratifin, also known as 14-3-3 sigma protein, stimulates collagenase (MMP-1) expression in dermal fibroblasts. However, role of extracellular stratifin in regulation of extracellular matrix (ECM) factors and other matrix metalloproteinases (MMPs) in dermal fibroblast remains unexplored. To address this question, large-scale ECM gene expression profile were analyzed in human dermal fibroblasts co-cultured with keratinocytes or treated with recombinant stratifin. Superarray pathway-specific microarrays were utilized to identify upregulation or downregulation of 96 human ECM and adhesion molecule genes. RT-PCR and Western blot were used to validate microarray expression profiles of selected genes. Comparison of gene profiles with the appropriate controls showed a significant (more than twofold) increase in expression of collagenase-1, stromelysin-1 and -2, neutrophil collagenase, and membrane type 5 MMP in dermal fibroblasts treated with stratifin or co-cultured with keratinocytes. Expression of type I collagen and fibronectin genes decreased in the same fibroblasts. The results of a dose-response experiment showed that stratifin stimulates the expression of stromelysin-1 (MMP-3) mRNA by dermal fibroblasts in a concentration-dependent fashion. Furthermore, Western blot analysis of fibroblast-conditioned medium showed a peak in MMP-3 protein levels 48 h following treatment with recombinant stratifin. In a lasting-effect study, MMP-3 protein was detected in fibroblast-condition medium for up to 72 h post removal of stratifin. In conclusion, our results suggest that keratinocyte-releasable stratifin plays a major role in induction of ECM degradation by dermal fibroblasts through stimulation of key MMPs, such as MMP-1 and MMP-3. Therefore, stratifin protein may prove to be a useful target for clinical intervention in controlling excessive wound healing in fibrotic conditions.

Introducing forage in dairy calf diets during the preweaning period stimulates development of rumen capacity and function and gut wall integrity. It was hypothesized that calves fed alfalfa hay (AH) at greater levels (up to 25% of DM) would result in an increase in starter diet intake and growth performance. The objective of this study was to determine the effects of increasing levels of supplemental AH on growth performance, rumen fermentation, blood metabolites, and feeding behavior in intact male dairy calves. For this purpose, 45 Holstein male calves (3 d of age and 41 ± 2.5 kg of BW) were randomly assigned to 1 of the following 3 starter diets differing in their forage level: 1) diet without AH (control), 2) diet with 12.5% AH on a DM basis, and 3) diet with 25% AH on a DM basis. Supplementation of AH in the starter diets linearly increased total DMI and ADG during the postweaning ( < 0.01) and overall ( = 0.02) periods. Alfalfa hay supplementation linearly increased ( < 0.01) ruminal pH on d 35 and 70 of the study. The concentrations of total VFA ( = 0.85), acetate ( = 0.74), propionate ( = 0.93), and butyrate ( = 0.91) were not affected by treatments. Furthermore, forage supplementation tended (linearly, = 0.08) to increase the acetate to propionate ratio compared with the control calves. Blood β-hydroxybutyrate (BHBA) concentrations in the control calves were less ( = 0.03) than in the AH-fed calves on d 70 of the study. Among all the measured variables of skeletal growth, supplementation of AH in the starter diets linearly increased heart girth ( = 0.05) and abdominal girth ( < 0.01) on d 70 of the study. These results indicate that the feed intake, feeding behavior, rumen fermentation parameters, and blood BHBA concentration may be affected by rations differing in AH level so that providing calves with 25% AH can improve calf performance.

We previously reported reduced expression of Sox4 in metastatic melanoma and its role in suppression of cell migration and invasion through inhibition of NF-κB p50. Sox4 can also bind to the promoter sequence of Dicer, a miRNA biogenesis factor. Interestingly, altered expression of Dicer was also observed in cancers. However, the potential mechanisms which regulate Dicer expression and its potential significance in melanoma progression are unknown. Here we studied the regulation of Dicer expression by Sox4 and its role in suppression of melanoma invasion. Our data showed that Sox4 positively regulates Dicer expression by binding to its promoter sequences and enhancing its activity. We found that knockdown of Dicer enhances the matrigel invasion of melanoma cells by at least 2-fold. In addition, we revealed that overexpression of exogenous Dicer reverts the enhanced melanoma cell invasion upon Sox4 knockdown. Furthermore, we examined the expression of Dicer protein in a large set of melanocytic lesions (n=504) at different stages by tissue microarray and found that Dicer expression is inversely correlated with melanoma progression (P < 0.0001). Consistently, reduced Dicer expression was correlated with a poorer overall and disease-specific 5-year survival of patients (P = 0.015 and 0.0029, respectively). In addition, we found a significant correlation between expression of Sox4 and Dicer proteins in melanoma biopsies (P = 0.009), further indicating the regulation of Dicer expression by Sox4. Finally, we revealed that knockdown of Sox4 induces a major change in the expression pattern of miRNAs in melanoma cells, mainly due to reduced expression of Dicer. Our results pinpoint the regulation of Dicer expression by Sox4 in melanoma and the critical role of Dicer in suppression of melanoma invasion. Our findings on Sox4 regulated miRNA biogenesis pathway may aid toward the development of novel targeted therapeutic approaches for melanoma.