scholarly journals Near-infrared oligonucleotide duplex sensors for imaging rapidly activated transcription factors in vitro and in situ

2019 ◽  
Author(s):  
Alexei A. Bogdanov ◽  
Valeriy Metelev ◽  
Toloo Taghian ◽  
Ilya D. Solovyev ◽  
Anand T. N. Kumar ◽  
...  
Keyword(s):  
Transcription Factors ◽  
Near Infrared ◽  
Oligonucleotide Duplex

Potential of solubility, enzymatic methods and NIRS to predict in situ rumen escape protein

1997 ◽  
Vol 45 (2) ◽  
pp. 291-306 ◽  
Author(s):  
J.L. De Boever ◽  
B.G. Cottyn ◽  
J.M. Vanacker ◽  
C.V. Boucque
Keyword(s):  
Phosphate Buffer ◽  
Near Infrared ◽  
Streptomyces Griseus ◽  
Maize Silage ◽  
Near Infrared Reflectance ◽  
In Vitro Method ◽  
Rumen Degradation ◽  
The Difference

The percentage of feed protein escaping rumen degradation was measured by the in situ method (%EPsitu) for 29 compound feeds, untreated and formaldehyde-treated soyabean meal and 12 forages: 3 grass silages, 2 maize silages, fresh grass, grass hay, fodder beets, fresh potatoes, ensiled beet pulp, chopped ear-maize silage and brewers' grains. Loss of particles through bag pores was determined by the difference between the washable fraction (W) and the fraction soluble in borate-phosphate buffer at pH 6.7 (S). W - S was most pronounced for compound feeds (on average 14.4 percentage units), for brewers' grains and maize silages. A correction of %EPsitu, assuming that W - S degrades like the potentially degradable fraction, was not appropriate. Solubility in borate-phosphate buffer after 1 h, enzymic degradability by protease from Streptomyces griseus or ficin after 1, 6 and 24 h and near infrared reflectance spectroscopy (NIRS) (for compound feeds alone) were examined as a routine method to predict %EPsitu. With the buffer and S. griseus the effect of pH (6.7 vs. 8.0) and at pH 8.0 the effect of amount of substrate (500-mg sample vs. 20 mg N) were tested. With ficin, 500-mg samples were incubated at pH 6.7. Predictions were better when compound feeds and forages were considered separately. However, the best in vitro method was different for the 2 feed categories, being solubility in buffer for the compound feeds and enzymic degradation of a constant amount of protein with S. griseus at pH 8.0 for forages. NIRS showed potential to predict %EPsitu of compound feeds, but needs more reference samples. The Dutch feed tables appeared more accurate than the best in vitro method for compound feeds, but was too inaccurate for some forages like fodder beets, maize silage and ear-maize silage.

scholarly journals Application of FTIR and Raman Spectroscopy to Characterisation of Bioactive Materials and Living Cells

2003 ◽  
Vol 17 (2-3) ◽  
pp. 275-288 ◽  
Author(s):  
I. Notingher ◽  
J. R. Jones ◽  
S. Verrier ◽  
I. Bisson ◽  
P. Embanga ◽  
...  
Keyword(s):  
Raman Spectroscopy ◽  
Near Infrared ◽  
Cell Damage ◽  
Living Cells ◽  
Biologically Active ◽  
Bioactive Materials ◽  
45S5 Bioglass ◽  
Ftir And Raman Spectroscopy

Both Fourier Transform Infrared (FTIR) and Raman spectroscopy have been applied to thein vitrocharacterisation of biomaterials, mainly surface reactions leading to the formation of a biologically active hydroxycarbonate apatite (HCA) layer on the sample surface when immersed in simulated body fluids (SBF). The HCA layer indicates the degree of bioactivity of the sample, because it leads to a strong bond between the biomaterial and living tissue. Reflection measurements using FTIR allow quick, non-destructive detection of the HCA layer for solid and powder samples. Due to the low Raman scattering efficiency and low absorption of water in the visible-near infrared region, Raman micro-spectroscopy was successfully used for thein situcharacterisation of 20 and 40µm diameter 45S5 Bioglass®fibres. Thein situcapabilities of the Raman micro-spectrometer have also been extended to the characterisation of living cells attached on bioinert silica and bioactive 45S5 Bioglass®and 58S substrates. Using a high power 785 nm laser, living cells in physiological conditions can be real-time sampled over long periods of time without inducing cell damage and with good signal strength. Cell death can be monitored because it proved to induce strong changes in the Raman signature in the spectral regions 1000–1150 cm–1and 1550–1650 cm–1.

scholarly journals Co-delivery of nanoparticle and molecular drug by hollow mesoporous organosilica for tumor-activated and photothermal-augmented chemotherapy of breast cancer

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Haixian Zhang ◽  
Feifei Song ◽  
Caihong Dong ◽  
Luodan Yu ◽  
Cai Chang ◽  
...  
Keyword(s):  
Tumor Microenvironment ◽  
High Performance ◽  
Near Infrared ◽  
Copper Ions ◽  
Therapeutic Modality ◽  
Mesoporous Organosilica ◽  
Hollow Interior

Abstract Background In comparison with traditional therapeutics, it is highly preferable to develop a combinatorial therapeutic modality for nanomedicine and photothermal hyperthermia to achieve safe, efficient, and localized delivery of chemotherapeutic drugs into tumor tissues and exert tumor-activated nanotherapy. Biocompatible organic–inorganic hybrid hollow mesoporous organosilica nanoparticles (HMONs) have shown high performance in molecular imaging and drug delivery as compared to other inorganic nanosystems. Disulfiram (DSF), an alcohol-abuse drug, can act as a chemotherapeutic agent according to its recently reported effectiveness for cancer chemotherapy, whose activity strongly depends on copper ions. Results In this work, a therapeutic construction with high biosafety and efficiency was proposed and developed for synergistic tumor-activated and photothermal-augmented chemotherapy in breast tumor eradication both in vitro and in vivo. The proposed strategy is based on the employment of HMONs to integrate ultrasmall photothermal CuS particles onto the surface of the organosilica and the molecular drug DSF inside the mesopores and hollow interior. The ultrasmall CuS acted as both photothermal agent under near-infrared (NIR) irradiation for photonic tumor hyperthermia and Cu2+ self-supplier in an acidic tumor microenvironment to activate the nontoxic DSF drug into a highly toxic diethyldithiocarbamate (DTC)-copper complex for enhanced DSF chemotherapy, which effectively achieved a remarkable synergistic in-situ anticancer outcome with minimal side effects. Conclusion This work provides a representative paradigm on the engineering of combinatorial therapeutic nanomedicine with both exogenous response for photonic tumor ablation and endogenous tumor microenvironment-responsive in-situ toxicity activation of a molecular drug (DSF) for augmented tumor chemotherapy. Graphical abstract

Correlated Studies of Cellular Interactions Using TEM, Freeze Etching, and SEM

1974 ◽  
Vol 32 ◽  
pp. 320-321
Author(s):  
J. P. Revel
Keyword(s):  
Developmental Stages ◽  
Three Dimensional ◽  
Cell Movement ◽  
Cellular Interactions ◽  
Serial Sections ◽  
Cell Sheets ◽  
Freeze Etching ◽  
Early Developmental

Movement of individual cells or of cell sheets and complex patterns of folding play a prominent role in the early developmental stages of the embryo. Our understanding of these processes is based on three- dimensional reconstructions laboriously prepared from serial sections, and from autoradiographic and other studies. Many concepts have also evolved from extrapolation of investigations of cell movement carried out in vitro. The scanning electron microscope now allows us to examine some of these events in situ. It is possible to prepare dissections of embryos and even of tissues of adult animals which reveal existing relationships between various structures more readily than used to be possible vithout an SEM.

In vitro and in vivo polysaccharides assembly: ultrastructural and cytochemical study of growing plant cell wall components.

1978 ◽  
Vol 36 (2) ◽  
pp. 434-435
Author(s):  
D. Reis ◽  
B. Vian ◽  
J. C. Roland
Keyword(s):  
Cell Wall ◽  
Self Assembly ◽  
Plant Cell Wall ◽  
Higher Plants ◽  
Three Dimensional ◽  
Cytochemical Study ◽  
Wall Components

Wall morphogenesis in higher plants is a problem still open to controversy. Until now the possibility of a transmembrane control and the involvement of microtubules were mostly envisaged. Self-assembly processes have been observed in the case of walls of Chlamydomonas and bacteria. Spontaneous gelling interactions between xanthan and galactomannan from Ceratonia have been analyzed very recently. The present work provides indications that some processes of spontaneous aggregation could occur in higher plants during the formation and expansion of cell wall.Observations were performed on hypocotyl of mung bean (Phaseolus aureus) for which growth characteristics and wall composition have been previously defined.In situ, the walls of actively growing cells (primary walls) show an ordered three-dimensional organization (fig. 1). The wall is typically polylamellate with multifibrillar layers alternately transverse and longitudinal. Between these layers intermediate strata exist in which the orientation of microfibrils progressively rotates. Thus a progressive change in the morphogenetic activity occurs.

In Situ Localization of PPAR Gamma and Uncoupling Protein in Mouse Embryo Sections Using Digoxigenin-Labeled Riboprobes

1996 ◽  
Vol 54 ◽  
pp. 32-33
Author(s):  
C. Jennermann ◽  
S. A. Kliewer ◽  
D. C. Morris
Keyword(s):  
Alkaline Phosphatase ◽  
Room Temperature ◽  
Uncoupling Protein ◽  
Ppar Gamma ◽  
Nuclear Hormone Receptor ◽  
Full Length ◽  
Northern Analysis ◽  
Peroxisome Proliferator

Peroxisome proliferator-activated receptor gamma (PPARg) is a member of the nuclear hormone receptor superfamily and has been shown in vitro to regulate genes involved in lipid metabolism and adipocyte differentiation. By Northern analysis, we and other researchers have shown that expression of this receptor predominates in adipose tissue in adult mice, and appears first in whole-embryo mRNA at 13.5 days postconception. In situ hybridization was used to find out in which developing tissues PPARg is specifically expressed.Digoxigenin-labeled riboprobes were generated using the Genius™ 4 RNA Labeling Kit from Boehringer Mannheim. Full length PPAR gamma, obtained by PCR from mouse liver cDNA, was inserted into pBluescript SK and used as template for the transcription reaction. Probes of average size 200 base pairs were made by partial alkaline hydrolysis of the full length transcripts. The in situ hybridization assays were performed as described previously with some modifications. Frozen sections (10 μm thick) of day 18 mouse embryos were cut, fixed with 4% paraformaldehyde and acetylated with 0.25% acetic anhydride in 1.0M triethanolamine buffer. The sections were incubated for 2 hours at room temperature in pre-hybridization buffer, and were then hybridized with a probe concentration of 200μg per ml at 70° C, overnight in a humidified chamber. Following stringent washes in SSC buffers, the immunological detection steps were performed at room temperature. The alkaline phosphatase labeled, anti-digoxigenin antibody and detection buffers were purchased from Boehringer Mannheim. The sections were treated with a blocking buffer for one hour and incubated with antibody solution at a 1:5000 dilution for 2 hours, both at room temperature. Colored precipitate was formed by exposure to the alkaline phosphatase substrate nitrobluetetrazoliumchloride/ bromo-chloroindlylphosphate.

CONSERVATION OF RARE SPECIES PLANTS AND LICHENS IN SITU WITH PLANNED ANTHROPOGENIC ACTIVITIES: BASIC APPROACHES AND IMPLEMENTATION PRINCIPLES

2018 ◽  
Vol 12 (7-8) ◽  
pp. 38-45
Author(s):  
A. N. EFREMOV ◽  
N. V. PLIKINA ◽  
T. ABELI
Keyword(s):  
Rare Species ◽  
Technology Implementation ◽  
Technology Development ◽  
Anthropogenic Activities ◽  
Natural Habitat ◽  
Local Population ◽  
Main Stage ◽  
Social Risks

Rare species are most vulnerable to man-made impacts, due to their biological characteristics or natural resource management. As a rule, the economic impact is associated with the destruction and damage of individual organisms, the destruction or alienation of habitats. Unfortunately, the conservation of habitat integrity is an important protection strategy, which is not always achievable in the implementation of industrial and infrastructural projects. The aim of the publication is to summarize the experience in the field of protection of rare species in the natural habitat (in situ), to evaluate and analyze the possibility of using existing methods in design and survey activities. In this regard, the main methodological approaches to the protection of rare species in the natural habitat (in situ) during the proposed economic activity were reflected. The algorithm suggested by the authors for implementing the in situ project should include a preparatory stage (initial data collection, preliminary risk assessments, technology development, obtaining permitting documentation), the main stage, the content of which is determined by the selected technology and a long monitoring stage, which makes it possible to assess the effectiveness of the taken measures. Among the main risks of in situ technology implementation, the following can be noted: the limited resources of the population that do not allow for the implementation of the procedure without prior reproduction of individuals in situ (in vitro); limited knowledge of the biology of the species; the possibility of invasion; the possibility of crossing for closely related species that сo-exist in the same habitat; social risks and consequences, target species or population may be important for the local population; financial risks during the recovery of the population. The available experience makes it possible to consider the approach to the conservation of rare species in situ as the best available technology that contributes to reducing negative environmental risks.

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