Quantitative analysis of myofilament protein phosphorylation in small cardiac biopsies

Phosphorylation of cardiac myofilament proteins represents one of the main post‐translational mechanisms that regulate cardiac pump function. Human studies are often limited by the amount of available tissue as biopsies taken during cardiac catheterization weigh only 1 mg (dry weight). Similarly, investigation of time‐ (or dose‐) dependent changes in protein phosphorylation in animal studies is often hampered by tissue availability. The present study describes quantitative analysis of phosphorylation status of multiple myofilament proteins by 2‐DE and Pro‐Q® Diamond stained gradient gels using minor amounts (?0.5 mg dry weight) of human and pig cardiac tissue.     

Effect of adhesive type and composite viscosity on the dentin bond strength

Objectives: Evaluate the influence of composite resins viscosity and type of cure of the adhesive systems on the bond strength of composite resins submitted to artificial aging.

Methods: Dentin specimens (n = 240) were divided into 2 groups: Group GC: GrandioSO, and Group GF: GrandioSO Heavy Flow. These groups were subdivided into 6: FM: Futurabond M – light cured, FDCC: Futurabond Dual Cure – chemical cured, FDCL: Futurabond Dual Cure – light cured, CS3: Clearfil S3 – light cured, CDCC: Clearfil Dual Cure – chemical cured, and CDCL: Clearfil Dual Cure – light cured. Resin blocks were build up on the dentin surface. Half of samples on each group were cut to obtain resin/dentin sticks (1 × 1 mm). The other half was first submitted to thermomechanical aging. The dentin/resin sticks were submitted to microtensile bond strength test and the results were analyzed using three-way ANOVA and Tukey’s test (α = 5%).

Results: ANOVA showed significant influence for adhesive (p = 0.0000) and aging (p = 0.0001). No significant influence of the composite viscosity on bond strength was observed (ANOVA: p = 0.0861). For adhesive, the results of Tukey’s test (MPa) were CDCC: 13.44 (±5.13)a; FM: 14,01 (±2.71)a; CDCL: 14.51 (±4.98)a; FDCC: 18.66 (±7.13)b; CS3: 18.80 (±6.50)b; FDCL: 19.18 (±7.39)b. For aging: AGED: 14.99 (±6.32)a; NOT AGED: 17.87 (±5.97)b.

Conclusion: Composite resin viscosities did not influence on the bond strength. Type of cure of the adhesives had influence on the bond strength. Thermomechanical aging decreased the bond strength.     

Effects of Rice Batter on Oil Uptake and Sensory Quality of Coated Fried Okra

ABSTRACT: Okra was coated and deep-fat fried with batters of flour sources including rice flour, a mixture of rice flour and small amounts of pregelatinized rice flour (PGRF), and, as a control, traditional wheat flour. The addition of PGRF, up to 8%, enhanced batter viscosity and the coating properties of the rice batter. Oil uptake of the fried batter decreased with the addition of up to 5% PGRF. Rice flour fried batters, with and without PGRF, were found to absorb substantially lower oil, by as much as 51 %, compared with the wheat batter. The fried okra coated with the rice batter containing 5% PGRF, when evaluated for sensory properties on appearance and surface attributes, was found to be superior or equal to those with the wheat batter and rice batter without PGRF. Particularly, its golden brown color is considered more desirable than the lighter yellow color of the other 2 entities. Similarly, most of its 1st-bite and after-chew properties were slightly better and were in the normal range of commercially available products. Specifically, its distinctive crispiness is considered a positive attribute, whereas its slightly higher tooth packing properties, while remaining in the range of commercial products, may be noticeable to some consumers.     

Fibroblast Activation Protein Targeted Photodynamic Therapy Selectively Kills Activated Skin Fibroblasts from Systemic Sclerosis Patients and Prevents Tissue Contraction

Systemic sclerosis (SSc) is a rare, severe, auto-immune disease characterized by inflammation, vasculopathy and fibrosis. Activated (myo)fibroblasts are crucial drivers of this fibrosis. By exploiting their expression of fibroblast activation protein (FAP) to perform targeted photodynamic therapy (tPDT), we can locoregionally deplete these pathogenic cells. In this study, we explored the use of FAP-tPDT in primary skin fibroblasts from SSc patients, both in 2D and 3D cultures. Method: The FAP targeting antibody 28H1 was conjugated with the photosensitizer IRDye700DX. Primary skin fibroblasts were obtained from lesional skin biopsies of SSc patients via spontaneous outgrowth and subsequently cultured on plastic or collagen type I. For 2D FAP-tPDT, cells were incubated in buffer with or without the antibody-photosensitizer construct, washed after 4 h and exposed to λ = 689 nm light. Cell viability was measured using CellTiter Glo^®®. For 3D FAP-tPDT, cells were seeded in collagen plugs and underwent the same treatment procedure. Contraction of the plugs was followed over time to determine myofibroblast activity. Results: FAP-tPDT resulted in antibody-dose dependent cytotoxicity in primary skin fibroblasts upon light exposure. Cells not exposed to light or incubated with an irrelevant antibody-photosensitizer construct did not show this response. FAP-tPDT fully prevented contraction of collagen plugs seeded with primary SSc fibroblasts. Even incubation with a very low dose of antibody (0.4 nM) inhibited contraction in 2 out of 3 donors. Conclusions: Here we have shown, for the first time, the potential of FAP-tPDT for the treatment of fibrosis in SSc skin.     

Porous alumina scaffolds chemically modified by calcium phosphate minerals and their application in bone grafts

The chemical modification of porous ceramic scaffold surfaces with calcium phosphate surges as an alternative to improve the bioactivity to be used as bone grafts. The biomimetic method has been commonly used to modify surfaces of Ti alloys but surges as alternative to modify ceramic biomaterials. Herein, we modified the surface of Al₂O₃ scaffolds with calcium phosphate minerals and strontium using the biomimetic method. The scaffolds were chemically treated using H₃PO₄ solution and then immersed in simulated body fluid 5× solution for 14 days. For the incorporation of strontium, they were immersed in an aqueous solution of 100 ppm analytical-grade Sr(NO₃)₂ under magnetic stirring. The samples were characterized by scanning electron microscopy, X-ray microtomography, X-ray diffraction, near-infrared spectroscopy, inductively coupled plasma emission spectroscopy, and energy-dispersive X-ray spectroscopy. The biocompatibility and ability to differentiate osteoblasts in vitro were evaluated using human cells. The incorporation of strontium into the phosphate structure was verified. Scaffolds were obtained with high porosity, three-dimensional structures, and the preferential adhesion and maturation of osteoblastic cells, which are essential to promote bone regeneration in vivo.     

Ultrastructural Imaging of Salmonella–Host Interactions Using Super‐resolution Correlative Light‐Electron Microscopy of Bioorthogonal Pathogens

The imaging of intracellular pathogens inside host cells is complicated by the low resolution and sensitivity of fluorescence microscopy and by the lack of ultrastructural information to visualize the pathogens. Herein, we present a new method to visualize these pathogens during infection that circumvents these problems: by using a metabolic hijacking approach to bioorthogonally label the intracellular pathogen Salmonella Typhimurium and by using these bioorthogonal groups to introduce fluorophores compatible with stochastic optical reconstruction microscopy (STORM) and placing this in a correlative light electron microscopy (CLEM) workflow, the pathogen can be imaged within its host cell context Typhimurium with a resolution of 20 nm. This STORM‐CLEM approach thus presents a new approach to understand these pathogens during infection.     

Use of Mild Heat Pre-treatment for Quality Retention of Fresh-cut Cantaloupe Melon

ABSTRACT: The effect on sensory attributes and shelf life of fresh-cut cantaloupe melon subjected to pre-cut heat treatment at 50 °C for 60 min, followed by storage at 4 °C prior to cutting, and then storage at 10 °C for 8 d was determined. Heat treatment reduced the rate of respiration and moisture loss during storage of the cut fruit. The treatment also reduced total microbial count during the 1st storage d and prevented growth of lactic acid bacteria that occurred in untreated fruit after 8 d in storage. Sensory evaluations indicate that heat treatment increased intensities of desirable attributes such as fruity melon and sweet aromatic flavors, and reduced undesirable flavors such as musty, sour, bitter, chemical and fermented. The study suggests that heat treatment would be useful in improving shelf life of fresh-cut fruit.