Progress in experimental stroke and translational medicine could be accelerated by high-resolution in vivo imaging of disease progression in the mouse cortex. Here, we introduce optical microscopic methods that monitor brain injury progression using intrinsic optical scattering properties of cortical tissue. A multi-parametric Optical Coherence Tomography (OCT) platform for longitudinal imaging of ischemic stroke in mice, through thinned-skull, reinforced cranial window surgical preparations, is described. In the acute stages, the spatiotemporal interplay between hemodynamics and cell viability, a key determinant of pathogenesis, was imaged. In acute stroke, microscopic biomarkers for eventual infarction, including capillary non-perfusion, cerebral blood flow deficiency, altered cellular scattering, and impaired autoregulation of cerebral blood flow, were quantified and correlated with histology. Additionally, longitudinal microscopy revealed remodeling and flow recovery after one week of chronic stroke. Intrinsic scattering properties serve as reporters of acute cellular and vascular injury and recovery in experimental stroke. Multi-parametric OCT represents a robust in vivo imaging platform to comprehensively investigate these properties.
This study investigated the effects of an antimicrobial peptide (AMP), cLF36, on growth performance and the histophysiological changes of the intestine in E. coli-challenged broiler chickens. A total number of 360 day old male chicks were randomly assigned to 4 groups of 6 replicates as follows: T1) negative control diet based on corn-soybean meal without E. coli challenge and additives; T2) positive control diet based on corn-soybean meal and challenged with E. coli without any additives; T3) positive control diet challenged with E. coli and supplemented with 20 mg AMP (cLF36)/kg diet; T4) positive control diet challenged with E. coli and supplemented with 45 mg antibiotic (bacitracin methylene disalicylate)/kg diet. Results showed that T3 improved growth performance and the jejunal morphology of E. coli-challenged chickens similar to those of T4. While antibiotic non-selectively decreased the population of ileal bacteria, AMP increased the population of Lactobacillus spp. and decreased harmful bacteria in the ileum of E. coli-challenged chickens. Supplementing E. coli-challenged chickens with AMP improved the gene expression of immune cells and upregulated the expression of tight junction proteins compared to other challenged groups. In conclusion, although cLF36 beneficially affected growth performance and the intestinal morphology of E. coli-challenged chickens similar to those of the antibiotic group, this AMP drastically improved the intestinal microbiome, immune cells, and junctional proteins compared to other E. coli-challenged birds, and can be nominated as an alternative for growth promoter antibiotics.
This study assessed the effects of 3 commercial organic acid (OA) preparations on growth performance, intestinal morphology, cecal microbiology, and immunity of Escherichia coli K88-challenged (ETEC) broiler chickens. One thousand one-day-old male broiler chickens were divided into 8 treatments of 5 replicate pens: Negative control (NC) birds received a basal diet (BD) and were not challenged with ETEC; positive control (PC) birds fed the BD and challenged with ETEC; BD + 0.2% (S1) or 0.4% (S2) of an OA mixture (Salkil) from one to 35 d; BD + 0.1, 0.075, and 0.05% (O1) of another OA mixture (Optimax) in the starter (one to 10 d), grower (11 to 24 d), and finisher (25 to 35 d) diets, respectively, or 0.1% (O2) from one to 35 d; BD + 0.07, 0.05, and 0.05% (P1) or 0.1, 0.07, and 0.05% (P2) of a further OA mixture (pHorce) in the starter, grower, and finisher diets, respectively. All groups (not NC) were challenged with one mL of ETEC (1 × 108 cfu/mL) at 7 d of age. The 3 OA mixtures are commercial formic and propionic acid preparations. Birds challenged with ETEC (PC) had reduced (P < 0.05) growth performance, ileal morphological parameters (not crypt depth, which was increased), cecal lactobacilli, and immune responses, and increased cecal E. coli compared with unchallenged, NC birds. The addition of OA to the diets of ETEC challenged birds (S1-P2) either numerically or significantly (P < 0.05) improved growth performance, ileal morphology and immune responses, increased cecal lactobacilli, and reduced cecal E. coli. For most OA additions, the assessed parameters were generally enhanced to equivalence to NC birds. The results suggest that dietary OA supplementation can enhance the growth performance, ileal morphology, cecal microbiota, and immunity of ETEC-challenged broilers to an extent that, under such circumstances, the formulations used in this study provided similar performance and assessed parameters as non-challenged birds.
This paper describes a novel optical method for label-free quantitative imaging of cerebral blood flow (CBF) and intracellular motility (IM) in the rodent cerebral cortex. This method is based on a technique that integrates dynamic light scattering (DLS) and optical coherence tomography (OCT), named DLS-OCT. The technique measures both the axial and transverse velocities of CBF, whereas conventional Doppler OCT measures only the axial one. In addition, the technique produces a three-dimensional map of the diffusion coefficient quantifying nontranslational motions. In the DLS-OCT diffusion map, we observed high-diffusion spots, whose locations highly correspond to neuronal cell bodies and whose diffusion coefficient agreed with that of the motion of intracellular organelles reported in vitro in the literature. Therefore, the present method has enabled, for the first time to our knowledge, label-free imaging of the diffusion-like motion of intracellular organelles in vivo. As an example application, we used the method to monitor CBF and IM during a brief ischemic stroke, where we observed an induced persistent reduction in IM despite the recovery of CBF after stroke. This result supports that the IM measured in this study represent the cellular energy metabolism-related active motion of intracellular organelles rather than free diffusion of intracellular macromolecules.
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