Contrasting acclimation mechanisms of berry color variant grapevine cultivars (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L. cv. Furmint) to natural sunlight conditions

The acclimation mechanisms of two berry color variant grapevine leaves, Furmint White (FW) and Furmint Red (FR), to natural sunlight conditions were investigated comparing leaves from two distinct locations: at canopy surface (sun-exposed leaves) and in the inner layers (shaded leaves). We found that in contrast to FR leaves, sun-exposed FW leaves were thicker than shaded leaves due to thicker palisade tissues. Confocal laser scanning microscopy of Naturstoff-treated leaf segments revealed that flavonoids were accumulated in nuclei, cell walls, cytoplasm, and chloroplasts of the adaxial epidermal and palisade layers of sun-exposed leaves in both cultivars. High-performance liquid chromatography analysis showed that the main phenolic components in both cultivars were caftaric acid and various glycosylated flavonols. Among the latter, the dominant component was quercetin glucuronide in both cultivars, unaffected by light conditions. However, caftaric acid and quercetin glucoside were present in significantly higher amounts in sun-exposed than in shaded leaves of both cultivars, but the effect of light conditions on caftaric acid contents was more pronounced in FR than in FW. Accordingly, the total polyphenol content of leaf extracts characterized by Folin-reagent reactivity was more enhanced in sun-exposed leaves of FR, than in FW. Our data suggest two different sunlight acclimation strategies to protect photosynthetic mesophyll tissues from potential photo-oxidative damage. One is realized in FW leaves as stronger shading by thicker palisade layer accompanied by enhanced chemical defense. The other is achieved in FR leaves via a more pronounced increase in caftaric acid and total polyphenol content but without morphological adjustments.     

Autophagy-dependent survival is controlled with a unique regulatory network upon various cellular stress events

Although autophagy is a type of programmed cell death, it is also essential for cell survival upon tolerable level of various stress events. For the cell to respond adequately to an external and/or internal stimulus induced by cellular stress, autophagy must be controlled in a highly regulated manner. By using systems biology techniques, here we explore the dynamical features of autophagy induction. We propose that the switch-like characteristic of autophagy induction is achieved by a control network, containing essential feedback loops of four components, so-called autophagy inducer, autophagy controller, mTORC1 and autophagy executor, respectively. We show how an autophagy inducer is capable to turn on autophagy in a cellular stress-specific way. The autophagy controller acts as a molecular switch and not only promotes autophagy but also blocks the permanent hyperactivation of the process via downregulating the autophagy inducer. In this theoretical analysis, we explore in detail the properties of all four proposed controlling elements and their connections. Here we also prove that the kinetic features of this control network can be considered accurate in various stress processes (such as starvation, endoplasmic reticulum stress and oxidative stress), even if the exact components may be different. The robust response of the resulting control network is essential during cellular stress.Subject terms: Biochemistry, Molecular biology     

Simplified process for the production of anti–CD19-CAR–engineered T cells

Background aimsAdoptive immunotherapy with the use of chimeric antigen receptor (CAR)-engineered T cells specific for CD19 has shown promising results for the treatment of B-cell lymphomas and leukemia. This therapy involves the transduction of autologous T cells with a viral vector and the subsequent cell expansion. We describe a new, simplified method to produce anti-CD19-CAR T cells.MethodsT cells were isolated from peripheral blood mononuclear cell (PBMC) with anti-CD3/anti-CD28 paramagnetic beads. After 2 days, the T cells were added to culture bags pre-treated with RetroNectin and loaded with the retroviral anti-CD19 CAR vector. The cells, beads and vector were incubated for 24 h, and a second transduction was then performed. No spinoculation was used. Cells were then expanded for an additional 9 days.ResultsThe method was validated through the use of two PBMC products from a patient with B-cell chronic lymphoblastic leukemia and one PBMC product from a healthy subject. The two PBMC products from the patient with B-cell chronic lymphoblastic leukemia contained 11.4% and 12.9% T cells. The manufacturing process led to final products highly enriched in T cells with a mean CD3+ cell content of 98%, a mean expansion of 10.6-fold and a mean transduction efficiency of 68%. Similar results were obtained from the PBMCs of the first four patients with acute lymphoblastic leukemia treated at our institution.ConclusionsWe developed a simplified, semi-closed system for the initial selection, activation, transduction and expansion of T cells with the use of anti-CD3/anti-CD28 beads and bags to produce autologous anti-CD19 CAR–transduced T cells to support an ongoing clinical trial.     

G-Rich Triplex-Forming Oligodeoxynucleotides as Transcription Repressors

Abstract

The ability of (A,G)- and (G,T)-oligonucleotides to form triple-helices with a critical polypurine-polypyrimidine sequence of the c-Ki-ras promoter has been examined, together with their capacity to inhibit the expression of the chloramphenicol acetyl-transferase gene driven by the c-Ki-ras promoter in a transfected cellular system.     

Apoptotic cells represent a dynamic stem cell niche governing proliferation and tissue regeneration

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Immunohistochemical visualisation of the enteric nervous system architecture in the germ-free piglets

Journal of Molecular Histology - The enteric nervous system (ENS), considered as separate branch of the autonomic nervous system, is located throughout the length of the gastrointestinal tract as a...     

Influence of physical exercise after long-lasting hypodynamia on the morphological parameters of muscle fibers

The purpose of this research was morphometric ultrastructure evaluation of the fibers in muscles taking part in the flight of pigeons forced to exercise after a long period of hypodynamia. It was found that following physical exercise, after 12 and 18 months of mobility limitation, there appeared marked qualitative and quantitative changes: a diminution of the volume fraction and number of mitochondria, increase of smooth sarcoplasmic reticulum and sarcoplasm, and a significant decrease of the number of glycogen granules as compared with those after 18 months hypodynamia. The above described changes were more pronounced in the supracoracoideus than in the pectoralis muscle.     

Spectroscopic and electron micrographic studies on the repeat tetrapeptide of tropoelastin: (Val-Pro-Gly-Gly)n

The polytetrapeptide repeat of tropoelastin, Val₁-Pro₂-Gly₃-Gly₄, coacervates from aqueous solution as temperature increases. The coacervation is a cooperative process which is concentration dependent, moving the temperature profile of coacervation curves to lower temperature with high concentrations. Electron micrographs of uranyl acetate or uranyl acetate-oxalic acid negatively stained coacervated samples are characterized by filamentous structures whose center-to-center diameter is approximately 50 Å as measured with optical diffraction. Coacervation thus results in increased intermolecular order. Circular dichroism spectra in methanol and trifluoroethanol solutions are significantly different from the spectrum in water. Once coacervated, though, the polytetrapeptide gave a CD spectrum very similar to that seen in these organic solvents. The data support the point of view that coacervation is the result of hydrophobic association attending an inverse temperature transition.