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PloS One 2019Dextranol, a reduced dextran, prevents damage to stored dry protein samples that unmodified dextran would otherwise cause. Desiccation protectants (xeroprotectants) like...
Dextranol, a reduced dextran, prevents damage to stored dry protein samples that unmodified dextran would otherwise cause. Desiccation protectants (xeroprotectants) like the polysaccharide dextran are critical for preserving dried protein samples by forming a rigid glass that protects entrapped protein molecules. Stably dried proteins are important for maintaining critical information in clinical samples like blood serum as well as maintaining activity of biologic drug compounds. However, we found that dextran reacts with both dried serum proteins and lyophilized purified proteins during storage, producing high-molecular weight Amadori-product conjugates. These conjugates appeared in a matter of days or weeks when stored at elevated temperatures (37° or 45°C), but also appeared on a timescale of months when stored at room temperature. We synthesized a less reactive dextranol by reducing dextran's anomeric carbon from an aldehyde to an alcohol. Serum samples dried in a dextranol-based matrix protected the serum proteins from forming high-molecular weight conjugates. The levels of four cancer-related serum biomarkers (prostate specific antigen, neuropilin-1, osteopontin, and matrix-metalloproteinase 7) decreased, as measured by immunoassay, when serum samples were stored for one to two weeks in dextran-based matrix. Switching to a dextranol-based xeroprotection matrix slightly reduced the damage to osteopontin and completely stopped any detectable damage during storage in the other three biomarkers when stored for a period of two weeks at 45°C. We also found that switching from dextran to dextranol in a lyophilization formulation eliminates this unwanted reaction, even at elevated temperatures. Dextranol offers a small and easy modification to dextran that significantly improves the molecule's function as a xeroprotectant by eliminating the potential for damaging protein-polysaccharide conjugation.
Topics: Blood Proteins; Desiccation; Dextrans; Drug Compounding; Oxidation-Reduction; Preservation, Biological; Protein Stability; Temperature
PubMed: 31490977
DOI: 10.1371/journal.pone.0222006 -
Journal of Clinical Microbiology Dec 2019Xpert MTB/RIF (Xpert) and culture are the most reliable methods for tuberculosis diagnosis but are still poorly accessible in many low-resource countries. We aimed to...
Xpert MTB/RIF (Xpert) and culture are the most reliable methods for tuberculosis diagnosis but are still poorly accessible in many low-resource countries. We aimed to assess the effects of OMNIgene Sputum (OM-S) and ethanol in preserving sputum for Xpert and OM-S for mycobacterial growth indicator tube (MGIT) testing over periods of 15 and 8 days, respectively. Sputum samples were collected from newly diagnosed smear-positive patients. For Xpert, pooled samples were split into 5 aliquots: 3 for Xpert on days 0, 7, and 15 without additive and 2 with either OM-S or ethanol at day 15. For MGIT, 2 aliquots were tested without preservative and 2 with OM-S at 0 and 8 days. Totals of 48 and 47 samples were included in the analysis for Xpert and culture. With Xpert, using day 0 as a reference, untreated samples stored for 7 and 15 days showed concordances of 45/46 (97.8%) and 46/48 (95.8%). For samples preserved with OM-S or ethanol for 15 days compared with untreated samples processed at day 0 or after 15 days, OM-S concordances were 46/48 (95.8%) and 47/48 (97.9%), while those of ethanol were 44/48 (91.7%) and 45/48 (93.8%). With MGIT, concordances between untreated and OM-S-treated samples were 21/41 (51.2%) at day 0 and 21/44 (47.7%) at day 8. In conclusion, Xpert equally detected tuberculosis in OM-S-treated and untreated samples up to 15 days but showed slightly lower detection in ethanol-treated samples. Among OM-S-treated samples, MGIT positivity was significantly lower than in untreated samples at both time points.
Topics: Bacteriological Techniques; Ethanol; Female; Humans; Indicators and Reagents; Male; Microbial Sensitivity Tests; Molecular Diagnostic Techniques; Mycobacterium tuberculosis; Preservation, Biological; Reproducibility of Results; Sensitivity and Specificity; Specimen Handling; Sputum; Tuberculosis; Uganda
PubMed: 31619525
DOI: 10.1128/JCM.00810-19 -
Science Advances Mar 2024Many biomolecular condensates are enriched in and depend on RNAs and RNA binding proteins (RBPs). So far, only a few studies have addressed the characterization of the...
Many biomolecular condensates are enriched in and depend on RNAs and RNA binding proteins (RBPs). So far, only a few studies have addressed the characterization of the intermolecular interactions responsible for liquid-liquid phase separation (LLPS) and the impact of condensation on RBPs and RNAs. Here, we present an approach to study protein-RNA interactions inside biomolecular condensates by applying cross-linking of isotope labeled RNA and tandem mass spectrometry to phase-separating systems (LLPS-CLIR-MS). LLPS-CLIR-MS enables the characterization of intermolecular interactions present within biomolecular condensates at residue-specific resolution and allows a comparison with the same complexes in the dispersed phase. We observe that sequence-specific RBP-RNA interactions present in the dispersed phase are generally maintained inside condensates. In addition, LLPS-CLIR-MS identifies structural alterations at the protein-RNA interfaces, including additional unspecific contacts in the condensed phase. Our approach offers a procedure to derive structural information of protein-RNA complexes within biomolecular condensates that could be critical for integrative structural modeling of ribonucleoproteins (RNPs) in this form.
Topics: Biomolecular Condensates; Preservation, Biological; Phase Separation; RNA; Ribonucleoproteins
PubMed: 38446881
DOI: 10.1126/sciadv.adm7435 -
The Journal of Parasitology Aug 1988The "true" coccidia (phylum Apicomplexa, suborder Eimeriina) constitute a large and heterogeneous group of parasitic protozoa. Despite the large number of described... (Review)
Review
The "true" coccidia (phylum Apicomplexa, suborder Eimeriina) constitute a large and heterogeneous group of parasitic protozoa. Despite the large number of described species (ca. 1,650) and the medical and veterinary importance of some (e.g., Toxoplasma), 2 facts are clear: (1) the majority of coccidia species are probably yet undescribed, and (2) the phylogenetic relationships of those described species are poorly known. Contributing to the latter dilemma is the lack of a tradition to provide type specimens by those who describe new species, even though the International Code of Zoological Nomenclature specifically recommends the designation of a type specimen with the description of a new species. With the publication of a new edition of the Code (1985), explicit provisions are made for the unique concerns of taxonomists working with Protozoa. Here we remind those interested in the taxonomy of coccidia of an already established method for preserving oocysts in resin and, as an alternative, suggest the standardization of a photographic procedure through which type specimens of coccidia oocysts might also be submitted to and maintained in accredited museums. Thus, coccidia taxonomists should no longer have an excuse for their failure to designate types.
Topics: Animals; Coccidia; Photomicrography; Preservation, Biological; Terminology as Topic
PubMed: 3294365
DOI: No ID Found -
Current Protocols in Microbiology Dec 2019Shigella species, which are closely related to Escherichia coli, can easily be maintained and stored in the laboratory. This article includes protocols for preparation...
Shigella species, which are closely related to Escherichia coli, can easily be maintained and stored in the laboratory. This article includes protocols for preparation of routine growth conditions and media, for storage of the bacteria, and for monitoring of the presence of the virulence plasmid. © 2019 by John Wiley & Sons, Inc. Basic Protocol 1: Growth of S. flexneri from frozen stocks or agar stabs Basic Protocol 2: Growth of S. flexneri in rich liquid medium Alternate Protocol 1: Growth of S. flexneri in rich defined medium Alternate Protocol 2: Growth of S. flexneri in minimal medium Basic Protocol 3: Storage of S. flexneri in frozen stocks Alternate Protocol 3: Storage of S. flexneri in agar stabs.
Topics: Bacteriological Techniques; Culture Media; Plasmids; Preservation, Biological; Shigella
PubMed: 31816179
DOI: 10.1002/cpmc.93 -
Scientific Reports Jan 2024The introduction of fish skin as a biological dressing for treating burns and wounds holds great promise, offering an alternative to existing management strategies....
The introduction of fish skin as a biological dressing for treating burns and wounds holds great promise, offering an alternative to existing management strategies. However, the risk of disease transmission is a significant concern. Therefore, this study aimed to examine how established sterilization and preservation procedures affected fish skin grafts' microbiological and histological properties for long-term usage. Lyophilization of the fish skin graft followed by rehydration in normal saline for 15 min did not change the collagen content. Furthermore, gamma irradiation of the lyophilized fish skin graft at different lengths 5, 10, and 25 KGy showed a significant reduction in microbial growth (aerobic bacteria, aerobic yeasts, and fungi) at 15- and 30 days after the irradiation. However, exposure to 10 KGy was found to be the most effective intensity among the different gamma irradiation lengths since it preserved the collagen fiber content and intensity in the lyophilized fish skin grafts at 15- and 30 days after the irradiation. These findings provide efficient preservation and sterilization methods for long-term usage of the fresh Tilapia skin grafts used for biological dressings.
Topics: Animals; Skin Transplantation; Preservation, Biological; Freeze Drying; Collagen; Fishes; Sterilization; Ichthyosis, Lamellar
PubMed: 38218988
DOI: 10.1038/s41598-024-51608-4 -
Theriogenology Feb 2023Sperm preservation is an efficient technique used for the recovery, conservation, and management of some endangered fish species. The present study was conducted to...
Sperm preservation is an efficient technique used for the recovery, conservation, and management of some endangered fish species. The present study was conducted to explore how preservation time would affect sperm and spawning performance in the endangered delta smelt (Hypomesus transpacificus). Sperm were preserved with the modified Hanks balanced salt solution at 14.7-16.9 °C. The Kruskal-Wallis test of sperm parameters using OpenCASA plugin in ImageJ software showed that sperm (n = 33♂) had significantly higher velocity and motility within the first 5 s after activation than that of other time points, while sperm had the lowest velocity and motility after 3 min post activation (P < 0.001). The findings (n = 30♂) also showed fresh sperm had higher velocity and motility than preserved sperm, while the sperm preserved for over 24 h showed a significantly low performance (P < 0.001). The nonlinear mixed effects models of fertilization results (n = 14♂ × 70♀) indicated the fresh sperm and sperm preserved for 1 h had higher fertilization rates than other preservation times (P < 0.001). The hatching rate (n = 14♂ × 70♀) also showed the fresh sperm and sperm preserved for 3 min and 1 h had higher hatching rates than other preservation times (P < 0.001). Overall, the study showed the best sperm performance in delta smelt was found within the first 5 s post activation, and the best fertilization and hatching rates were found when the sperm were fresh and preserved for 1 h. The findings of this study provide information for the first time about how long the delta smelt's sperm are motile for quality analysis, and how the preservation time can affect sperm quality, fertility, and hatching of this species for future applications.
Topics: Male; Animals; Semen; Osmeriformes; Cryopreservation; Sperm Motility; Spermatozoa; Endangered Species; Semen Preservation
PubMed: 36542880
DOI: 10.1016/j.theriogenology.2022.11.029 -
Revista Argentina de Microbiologia 2017The collection of fungal pathogens and symbionts of insects and other arthropods of the Centro de Estudios Parasitológicos y de Vectores, La Plata, Argentina, is unique...
The collection of fungal pathogens and symbionts of insects and other arthropods of the Centro de Estudios Parasitológicos y de Vectores, La Plata, Argentina, is unique because it preserves in vivo and in vitro cultures of fungal pathogens. This culture collection is open for research, teaching, consulting services, and strain deposit. It contains 421 strains belonging to 23 genera (16 Ascomycota, 4 Entomophthoromycotina, 2 Glomeromycota and 1 Oomycota), and the cultures are preserved by different methods such as cryopreservation in freezer at -20°C and -70°C, paper, distilled water and lyophilization. Fungi were isolated from insects, other arthropods, and soil (by using insect baits and selective media). Species were identified by morphological features and in a few strains by molecular taxonomy (PCR of rDNA). This collection is a reference center for species identification/certifications, research and teaching purposes, strain deposit, transference and consultancy services, and its overall goal is to preserve the fungal germplasm and ex situ diversity. Most of the strains are native of Argentina. The collection was originated in 1988 and is registered in the Latin American Federation for Culture Collections and in the World Federation of Culture Collections.
Topics: Animals; Argentina; Arthropods; Ascomycota; Insecta; Preservation, Biological; Symbiosis
PubMed: 28320556
DOI: 10.1016/j.ram.2016.09.007 -
PeerJ 2022The transport and storage of samples in temperatures of minus 80 °C is commonly considered as the gold standard for microbiome studies. However, studies conducting...
BACKGROUND
The transport and storage of samples in temperatures of minus 80 °C is commonly considered as the gold standard for microbiome studies. However, studies conducting sample collection at remote sites without a reliable cold-chain would benefit from a sample preservation method that allows transport and storage at ambient temperature.
METHODS
In this study we compare alpha diversity and 16S microbiome composition of 20 fecal sample replicates from Damaraland mole-rats () preserved in a minus 80 °C freezer and transported on dry ice to freeze-dried samples that were stored and transported in ambient temperature until DNA extraction.
RESULTS
We found strong correlations between relative abundances of Amplicon Sequence Variants (ASVs) between preservation treatments of the sample, no differences in alpha diversity measures between the two preservation treatments and minor effects of the preservation treatment on beta diversity measures. Our results show that freeze-drying samples can be a useful method for cost-effective transportation and storage of microbiome samples that yields quantitatively almost indistinguishable results in 16S microbiome analyses as those stored in minus 80 °C.
Topics: Feces; Freeze Drying; Preservation, Biological; Microbiota; Refrigeration
PubMed: 35310158
DOI: 10.7717/peerj.13095 -
Sensors (Basel, Switzerland) Sep 2021The protection of artistic and cultural heritage is a major challenge due to its peculiarities and its exposure to significant natural hazards. Several methodologies...
The protection of artistic and cultural heritage is a major challenge due to its peculiarities and its exposure to significant natural hazards. Several methodologies exist to assess the condition of artistic heritage and to protect it from exceptional actions. Moreover, novel digital technologies offer many solutions able to deliver a digital replica of artifacts of interest, so that a reduction in the uncertainties in the analysis models can be achieved. A rational approach to the preservation and protection of artistic heritage is based on traditional approaches supported and integrated by novel technologies, so that qualitative and quantitative indicators of the current condition of artistic heritage can be defined and validated in an interdisciplinary framework. The present paper reports the results of an approach to the maintenance and preservation of art objects housed in a museum complex based on a comprehensive digital path towards a Historical Digital Twin (HDT). A workflow aimed at estimating the stress regime and the dynamic properties of two sculptures, based on the detailed three-dimensional model resulting from a laser scanner survey, is illustrated and discussed. The results highlight the great advantages resulting from the integration of traditional and novel procedures in the field of conservation of artistic assets.
Topics: Art; Artifacts; Lasers; Museums; Preservation, Biological
PubMed: 34502848
DOI: 10.3390/s21175956