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.

Effect of spray drying and storage on the stability of bayberry polyphenols

2011 ◽  
Vol 129 (3) ◽  
pp. 1139-1147 ◽  
Author(s):  
Zhongxiang Fang ◽  
Bhesh Bhandari
Keyword(s):  
Spray Drying ◽  
The Stability ◽  
And Storage

scholarly journals LIM domain-wide comprehensive mutagenesis reveals the role of leucine in CSRP3 protein stability

2021 ◽  
Author(s):  
Pankaj Kumar Chauhan ◽  
Ramanathan Sowdhamini
Keyword(s):  
Protein Structures ◽  
Screening Method ◽  
Md Simulations ◽  
Specific Protein ◽  
Lim Domain ◽  
Lim Domains ◽  
Screening Protocol ◽  
The Stability ◽  
Glycine Rich Protein

Cardiomyopathies are a severe and chronic cardiovascular burden worldwide, affecting a large cohort in the general population. Cysteine and glycine-rich protein 3 (CSRP3) is one of key proteins implicated in dominant dilated cardiomyopathy (DCM) and hypertrophic cardiomyopathy (HCM). In this study, we device a rapid in-silico screening protocol that creates a mutational landscape map for all possible allowed and disallowed substitutions in the protein of interest. This map provides the structural and functional insights on the stability of LIM domains of CSRP3. Further, the sequence analysis delineates the eukaryotic CSRP3 protein orthologs which complements the mutational map. Next, we also evaluated the effect of HCM/DCM mutations on these domains. One of highly destabilising mutations - L44P (also disease causing) and a neutral mutation - L44M were further subjected to molecular dynamics (MD) simulations. The results establish that L44P substitution affects the LIM domain structure. The present study provides a useful perspective to our understanding of the role of mutations in the CSRP3 LIM domains and their evolution. Experimentally verifying every reported mutation can become challenging both in time and resources used. This study provides a novel screening method for quick identification of key mutation sites for specific protein structures that can reduce the burden on experimental research.

scholarly journals LIM Domain-Wide Comprehensive Virtual Mutagenesis Provides Structural Rationale for Cardiomyopathy Mutations in CSRP3

2021 ◽  
Author(s):  
Pankaj Kumar Chauhan ◽  
R. Sowdhamini
Keyword(s):  
Protein Structures ◽  
Screening Method ◽  
Md Simulations ◽  
Specific Protein ◽  
Lim Domain ◽  
Lim Domains ◽  
Screening Protocol ◽  
The Stability ◽  
Cardiomyopathy Mutations

Abstract Cardiomyopathies are a severe and chronic cardiovascular burden worldwide, affecting a large cohort in the general population. Cysteine and glycine-rich protein 3 (CSRP3) is one of key proteins implicated in dominant dilated cardiomyopathy (DCM) and hypertrophic cardiomyopathy (HCM). In this study, we device a rapid in-silico screening protocol that creates a mutational landscape map for all possible allowed and disallowed substitutions in the protein of interest. This map provides the structural and functional insights on the stability of LIM domains of CSRP3. Further, the sequence analysis delineates the eukaryotic CSRP3 protein orthologs which complements the mutational map. Next, we also evaluated the effect of HCM/DCM mutations on these domains. One of highly destabilising mutations - L44P (also disease causing) and a neutral mutation - L44M were further subjected to molecular dynamics (MD) simulations. The results establish that L44P substitution affects the LIM domain structure. The present study provides a useful perspective to our understanding of the role of mutations in the CSRP3 LIM domains and their evolution. Experimentally verifying every reported mutation can become challenging both in time and resources used. This study provides a novel screening method for quick identification of key mutation sites for specific protein structures that can reduce the burden on experimental research.

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.

Pre-selection of protective colloids for enhanced viability of Bifidobacterium bifidum following spray-drying and storage, and evaluation of aguamiel as thermoprotective prebiotic

2007 ◽  
Vol 40 (10) ◽  
pp. 1299-1306 ◽  
Author(s):  
M.E. Rodríguez-Huezo ◽  
R. Durán-Lugo ◽  
L.A. Prado-Barragán ◽  
F. Cruz-Sosa ◽  
C. Lobato-Calleros ◽  
...  
Keyword(s):  
Spray Drying ◽  
Bifidobacterium Bifidum ◽  
And Storage ◽  
Selection Of

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.

Rapid Detection of Bifidobacterium dentium by Enzymatic Hydrolysis of β-Glucuronide Substrates

1992 ◽  
Vol 55 (4) ◽  
pp. 291-295 ◽  
Author(s):  
DENIS ROY ◽  
PIERRE WARD
Keyword(s):  
Rapid Detection ◽  
Screening Method ◽  
Selective Medium ◽  
Rapid Screening ◽  
Staining Procedure ◽  
Bifidobacterium Adolescentis ◽  
Glucuronidase Activity ◽  
Hydrolysis Of ◽  
Selection Of ◽  
Glucuronidase Assay

Enzyme profiles and fermentation patterns of bifidobacteria were studied to determine phenotypic characteristics that allow the rapid detection of Bifidobacterium dentium and its differentiation from Bifidobacterium adolescentis, Bifidobacterium angulatum, Bifidobacterium catenulatum, and Bifidobacterium pseudocatenulatum. Among 43 bifidobacterial strains tested, the production of β-glucuronidase was limited to six strains of B. dentium. The presence of B. dentium on a selective medium may be rapidly confirmed by the detection of β-glucuronidase activity. Columbia agar containing propionic acid was chosen to enumerate bifidobacteria previously cultivated in MRS medium. After 48 h of incubation, β-glucuronidase activity was determined by using a plate staining procedure. B. dentium strains gave positive results for β-glucuronidase activity after application of the overlay solution of β-glucuronide substrate. The β-glucuronidase assay is a rapid screening method for B. dentium. This method might be useful for selection of nonpathogenic strains or detection of fecal contamination from human origin.

Stability Study for 53 Antibiotics in Solution and in Fortified Biological Matrixes by LC/MS/MS

2013 ◽  
Vol 96 (2) ◽  
pp. 471-480 ◽  
Author(s):  
Murielle Gaugain ◽  
Marie-Pierre Chotard ◽  
Eric Verdon
Keyword(s):  
Method Development ◽  
Screening Method ◽  
Monitoring Program ◽  
Negative Control ◽  
Stability Study ◽  
Cow Milk ◽  
Antibiotic Residues ◽  
Time Saving ◽  
The Stability ◽  
And Storage

Abstract This study was performed in order to determine the stability of antibiotics belonging to eight families in solution or biological matrix. Knowledge of the stability of antibiotics has to be demonstrated during method development or validation. The stability of stock standard solutions of 53 antibiotics was assessed after determining the appropriate conditions of dissolution and storage. The stability of the same 53 antibiotics after addition to negative control cow milk or pork muscle tissue stored at –18 and –70°C was also assessed. Our concern was to obtain information concerning the stability of antibiotic residues in fortified biological matrixes in order to make easier the implementation of a routine screening method for antibiotic residues within the framework of the French monitoring program. Antibiotic solutions and fortified samples were analyzed using an LC/MS/MS method previously validated for screening purposes and for which it was checked that all pertinent criteria to obtain interpretable stability results were fulfilled. The design for testing the stability of antibiotics in solutions and matrix samples is described, as well as the rules of decision that were observed. Term periods for the stability study ranged from 1 month to 1 year, depending on the class of compounds. The results presented in this article will be useful and time-saving for many reference and field laboratories because LC/MS/MS methods are more and more commonly used for screening purposes.

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.

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