scholarly journals Implementation of Mass Cytometry for Immunoprofiling of Patients with Solid Tumors

2019 ◽  
Vol 2019 ◽  
pp. 1-10
Author(s):  
Ingrid Poláková ◽  
Ondřej Pelák ◽  
Daniel Thürner ◽  
Barbora Pokrývková ◽  
Ruth Tachezy ◽  
...  
Keyword(s):  
Sample Preparation ◽  
Immune Responses ◽  
Immune Cells ◽  
Tumor Tissue ◽  
Important Criterion ◽  
Mass Cytometry ◽  
The Stability ◽  
And Storage ◽  
Selection Of ◽  
Conventional Staging

Monitoring immune responses to solid cancers may be a better prognostic tool than conventional staging criteria, and it can also serve as an important criterion for the selection of individualized therapy. Multiparametric phenotyping by mass cytometry extended possibilities for immunoprofiling. However, careful optimization of each step of such method is necessary for obtaining reliable results. Also, with respect to procedure length and costs, sample preparation, staining, and storage should be optimized. Here, we designed a panel of 31 antibodies which allows for identification of several subpopulations of lymphoid and myeloid cells in a solid tumor and peripheral blood simultaneously. For sample preparation, disaggregation of tumor tissue with two different collagenases combined with DNase I was compared, and removal of dead or tumor cells by magnetic separation was evaluated. Two possible procedures of barcoding for single-tube staining of several samples were examined. While the palladium-based barcoding affected the stability of several antigens, the staining with two differently labeled CD45 antibodies was suitable for cells isolated from a patient’s blood and tumor. The storage of samples in the intercalation solution for up to two weeks did not influence results of the analysis, which allowed the measurement of samples collected within this interval on the same day. This procedure optimized on samples from patients with head and neck squamous cell carcinoma enabled identification of various immune cells including rare subpopulations.

Development of a rapid screening protocol for selection of strains resistant to spray drying and storage in dry powder

2010 ◽  
Vol 1 (2) ◽  
pp. 165-174 ◽  
Author(s):  
S. Reimann ◽  
F. Grattepanche ◽  
C. Baggenstos ◽  
E. Rezzonico ◽  
B. Berger ◽  
...  
Keyword(s):  
Spray Drying ◽  
Bifidobacterium Longum ◽  
Survival Rates ◽  
Screening Method ◽  
Rapid Screening ◽  
Fast Screening ◽  
Screening Protocol ◽  
The Stability ◽  
And Storage ◽  
Selection Of

An efficient screening method for selection of Bifidobacterium longum strains resistant to spray drying and storage was developed based on randomly amplified polymorphic DNA (RAPD) for identification of the best survivors in mixed strains bacterial preparations. Three different primers were used to generate RAPD profiles of 22 B. longum strains. All strains were distinguished according to their RAPD profiles except for the strain NCC2705 and its H2O2 resistant derivative variant. The 22 strains were grouped in 3 batches of 7, 7 and 8 strains and subjected to spray drying and storage at 30 and 37 °C under anaerobic conditions. Batch survival rates after spray drying reached 17.1±4.4%. Strains showing the highest prevalence and/or resistance to storage at 37 °C were selected from individual batches for subsequent spray drying and storage testing. After 67 days of storage, NCC572 was identified as the dominant strain in powder. The stability of strain NCC572 was confirmed by performing single spray drying and storage tests. Out of 22 B. longum strains, a robust strain was identified by combining RAPD with a simultaneous screening test for survival under spray drying and storage. The method allowed a fast screening of B. longum strains in mixture for resistance to spray drying and storage compared to traditional screening procedures carried out with individual strains, in the same conditions. This approach could be applied to other stress conditions.

Post-translational modifications and stabilization of microtubules regulate transport of viral factors during infections

2021 ◽  
Vol 49 (4) ◽  
pp. 1735-1748
Author(s):  
Silvia Requena ◽  
Francisco Sánchez-Madrid ◽  
Noa B. Martín-Cófreces
Keyword(s):  
Immune Responses ◽  
Immune Cells ◽  
Building Blocks ◽  
Cell Types ◽  
Post Translational Modifications ◽  
Cellular Processes ◽  
Associated Proteins ◽  
The Stability ◽  
Different Characteristics ◽  
Β Tubulin

Tubulin post-translational modifications (PTMs) constitute a source of diversity for microtubule (MT) functions, in addition to the different isotypes of α and β-tubulin acting as building blocks of MTs. Also, MT-associated proteins (MAPs) confer different characteristics to MTs. The combination of all these factors regulates the stability of these structures that act as rails to transport organelles within the cell, facilitating the association of motor complexes. All these functions are involved in crucial cellular processes in most cell types, ranging from spindle formation in mitosis to the defense against incoming cellular threats during phagocytosis mediated by immune cells. The regulation of MT dynamics through tubulin PTMs has evolved to depend on many different factors that act in a complex orchestrated manner. These tightly regulated processes are particularly relevant during the induction of effective immune responses against pathogens. Viruses have proved not only to hijack MTs and MAPs in order to favor an efficient infection, but also to induce certain PTMs that improve their cellular spread and lead to secondary consequences of viral processes. In this review, we offer a perspective on relevant MT-related elements exploited by viruses.

scholarly journals Thermokinetics and rheology of agarose gel applied to bioprinting technology

2020 ◽  
Vol 24 (1 Part A) ◽  
pp. 347-353
Author(s):  
Boris Pokusaev ◽  
Andrey Vyazmin ◽  
Nikolay Zakharov ◽  
Sergey Karlov ◽  
Dmitry Nekrasov ◽  
...  
Keyword(s):  
Rheological Properties ◽  
Point Of View ◽  
Gel Formation ◽  
Operating Modes ◽  
Preliminary Preparation ◽  
Immobilized Microorganisms ◽  
The Stability ◽  
And Storage ◽  
Kinetics Of ◽  
Selection Of

The paper presents new results on the study of thermokinetics of gel system based on agarose in the process of transition from solution to gel and opposite. This issue is extremely relevant, since the stability and predictability of thermophysical and rheological properties in such transformations, especially in the presence of components of the nutrient medium and immobilized microorganisms, come to the fore in terms of design and selection of modes of operation of the printing device promising 3-D bioprinters, as well as the system of preparation and storage of the presence of the hysteresis effect, both from the point of view of the kinetics of gel formation and from the point of view of the dependence of rheological properties on temperature, at different concentrations of modifying components, is shown. The obtained results allow to draw a conclusion about the possibility of using the scheme with preliminary preparation of the initial biogel for the implementation of bioprinting technology based on agarose, and to recommend the obtained values for modeling the operating modes of devices of this type.

scholarly journals Aptamer-assisted tumor localization of bacteria for enhanced biotherapy

2021 ◽  
Author(s):  
Zhongmin Geng ◽  
Zhenping Cao ◽  
Rui Liu ◽  
Ke Liu ◽  
Jinyao Liu ◽  
...  
Keyword(s):  
Immune Responses ◽  
Tumor Tissue ◽  
Feed Ratio ◽  
Tumor Bearing Mouse ◽  
Bacterial Surface ◽  
Tumor Bearing ◽  
Different Types ◽  
Specific Accumulation ◽  
The Stability

Abstract Despite bacterial-mediated biotherapies have been widely explored for treating different types of cancer, their implementation has been restricted by severe side effects and low treatment efficacies, due largely to the absence of tumor-specific accumulation following administration. Here, the conjugation of aptamers to bacterial surface is described by a simple and cytocompatible amidation procedure, which can significantly promote the localization of bacteria in tumor site after systemic administration. The surface density of aptamers can be easily adjusted by varying feed ratio and the conjugation is able to increase the stability of anchored aptamers. Optimal bacteria conjugated with an average of 2.8 × 105 aptamers per cell present the highest specificity to tumor cells in vitro, separately generating near 2- and 5-times higher accumulation in tumor tissue at 12 and 60 hours compared to unmodified bacteria. In both 4T1 and H22 tumor-bearing mouse models, aptamer-conjugated attenuated Salmonella show strikingly enhanced antitumor efficacy, along with highly activated immune responses inside the tumor. This work demonstrates how bacterial behaviors can be tuned by surface conjugation and supports the potential of aptamer-conjugated bacteria for both targeted intratumoral localization and enhanced tumor biotherapy.

scholarly journals Measurement of Plasma-Derived Substance P: Biological, Methodological, and Statistical Considerations

2006 ◽  
Vol 13 (11) ◽  
pp. 1197-1203 ◽  
Author(s):  
Donald E. Campbell ◽  
Nancy Raftery ◽  
Richard Tustin ◽  
Nancy B. Tustin ◽  
Michelle L. DeSilvio ◽  
...  
Keyword(s):  
Substance P ◽  
Sample Preparation ◽  
Immune Responses ◽  
Comparative Evaluation ◽  
Careful Consideration ◽  
Detection Methods ◽  
Immune Disorders ◽  
Selection Of

ABSTRACT The undecapeptide substance P (SP) is a member of the tachykinin family of neurotransmitters, which has a pivotal role in the regulation of inflammatory and immune responses. One of the major barriers to the study of the in vivo role of SP in a number of immune disorders is the accurate measurement of SP in fluids. This is reflected in the variability of reported SP levels in serum and plasma of humans in both healthy and diseased states. This study was initiated in order to identify sources of variability by the comparative evaluation of the influences of sample preparation and analytical detection methods on the measurement of SP in plasma. The results indicate that sample preparation (peptide extraction versus no extraction) and the choice of analytical method for SP quantitation may yield significantly different values and may contribute to the variability in SP values reported in the literature. These results further emphasize the need for careful consideration in the selection of methods for SP quantitation, as well as caution in the interpretation and comparison of data reported in the literature.

scholarly journals Aptamer-assisted tumor localization of bacteria for enhanced biotherapy

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Zhongmin Geng ◽  
Zhenping Cao ◽  
Rui Liu ◽  
Ke Liu ◽  
Jinyao Liu ◽  
...  
Keyword(s):  
Immune Responses ◽  
Tumor Tissue ◽  
Feed Ratio ◽  
Tumor Bearing Mouse ◽  
Bacterial Surface ◽  
Tumor Bearing ◽  
Different Types ◽  
Specific Accumulation ◽  
The Stability

AbstractDespite bacterial-mediated biotherapies have been widely explored for treating different types of cancer, their implementation has been restricted by low treatment efficacy, due largely to the absence of tumor-specific accumulation following administration. Here, the conjugation of aptamers to bacterial surface is described by a simple and cytocompatible amidation procedure, which can significantly promote the localization of bacteria in tumor site after systemic administration. The surface density of aptamers can be easily adjusted by varying feed ratio and the conjugation is able to increase the stability of anchored aptamers. Optimal bacteria conjugated with an average of 2.8 × 105 aptamers per cell present the highest specificity to tumor cells in vitro, separately generating near 2- and 4-times higher accumulation in tumor tissue at 12 and 60 hours compared to unmodified bacteria. In both 4T1 and H22 tumor-bearing mouse models, aptamer-conjugated attenuated Salmonella show enhanced antitumor efficacy, along with highly activated immune responses inside the tumor. This work demonstrates how bacterial behaviors can be tuned by surface conjugation and supports the potential of aptamer-conjugated bacteria for both targeted intratumoral localization and enhanced tumor biotherapy.

Author response for "Mass cytometry analysis of blood immune cells from psoriasis patients on biological therapy"

2020 ◽  
Author(s):  
SM Solberg ◽  
AK Aarebrot ◽  
I Sarkar ◽  
A Petrovic ◽  
LF Sandvik ◽  
...  
Keyword(s):  
Immune Cells ◽  
Biological Therapy ◽  
Author Response ◽  
Mass Cytometry

Preparation, Stabilization and Some Properties of Purified Human Plasmin*

1964 ◽  
Vol 11 (01) ◽  
pp. 075-084 ◽  
Author(s):  
Daniel L Kline ◽  
Jacob B Fishman ◽  
Keyword(s):  
Plasminogen Activator ◽  
Stable Form ◽  
Neutral Ph ◽  
The Stability ◽  
And Storage

Summary1. Lysine increased the solubility, decreased the SK requirement and increased the stability of plasmin prepared from purified plasminogen by SK activation.2. A procedure is presented for the rapid and quantitative conversion of plasminogen to plasmin and storage of the plasmin in stable form at neutral pH as a lyophilized powder.3. Approximately 10% for the plasminogen molecule was split off during its activation. No carbohydrate was lost.4. The plasmin isolated was homogeneous in the ultracentrifuge at pH 2.5 and was quantitatively convertible to plasminogen activator by the addition of SK.

Evaluation of the quality and the stability of human samples for analysis and storage in biorepository

2018 ◽  
Vol 50 (1) ◽  
Author(s):  
Leonardo André Lange ◽  
Caroline Galgowski ◽  
Anna Cecília Roncalio ◽  
Fabiana Sehnem ◽  
Grabriela Borgmann ◽  
...  
Keyword(s):  
Human Samples ◽  
The Stability ◽  
And Storage

scholarly journals The Impact of Alkaloid-Producing Epichloë Endophyte on Forage Ryegrass Breeding: A New Zealand Perspective

Toxins ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 158
Author(s):  
Colin Eady
Keyword(s):  
Seed Production ◽  
Health Issues ◽  
Abiotic Stressors ◽  
Livestock Health ◽  
The Right ◽  
The Impact ◽  
The Relationship ◽  
And Storage ◽  
Selection Of ◽  
Epichloë Endophyte

For 30 years, forage ryegrass breeding has known that the germplasm may contain a maternally inherited symbiotic Epichloë endophyte. These endophytes produce a suite of secondary alkaloid compounds, dependent upon strain. Many produce ergot and other alkaloids, which are associated with both insect deterrence and livestock health issues. The levels of alkaloids and other endophyte characteristics are influenced by strain, host germplasm, and environmental conditions. Some strains in the right host germplasm can confer an advantage over biotic and abiotic stressors, thus acting as a maternally inherited desirable ‘trait’. Through seed production, these mutualistic endophytes do not transmit into 100% of the crop seed and are less vigorous than the grass seed itself. This causes stability and longevity issues for seed production and storage should the ‘trait’ be desired in the germplasm. This makes understanding the precise nature of the relationship vitally important to the plant breeder. These Epichloë endophytes cannot be ‘bred’ in the conventional sense, as they are asexual. Instead, the breeder may modulate endophyte characteristics through selection of host germplasm, a sort of breeding by proxy. This article explores, from a forage seed company perspective, the issues that endophyte characteristics and breeding them by proxy have on ryegrass breeding, and outlines the methods used to assess the ‘trait’, and the application of these through the breeding, production, and deployment processes. Finally, this article investigates opportunities for enhancing the utilisation of alkaloid-producing endophytes within pastures, with a focus on balancing alkaloid levels to further enhance pest deterrence and improving livestock outcomes.

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