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Romanian Journal of Morphology and... 2012A sequence of technically reproducible procedures is mandatory to guarantee a proper preservation of tissues and to build up the basis for sound diagnoses. However,... (Review)
Review
A sequence of technically reproducible procedures is mandatory to guarantee a proper preservation of tissues and to build up the basis for sound diagnoses. However, while the goal of these procedures was, until recently, to assure only structural (histological and cytological) preservation, an appropriate preservation of antigenic properties and of nucleic acid integrity is now additionally requested, in order to permit pathologists to provide the biological information necessary for the adoption of personalized therapies. The present review analyses the sequence of technical steps open to critical variations. Passages such as dehydration, paraffin embedding, sectioning and staining are relatively well standardized and allow adoption of dedicated (automatic) apparatuses, while other pre-analytical steps, i.e. time and modalities of transfer of surgical specimens from the surgical theatre to the pathology laboratory (s.c. "ischemia time") and the type and length of fixation are not standardized and are a potential cause of discrepancies in diagnostic results. Our group is involved in European-funded projects tackling these problems with the concrete objective of implementing a model of effective tumors investigations by high performance genetic and molecular methodologies. The problem of the discrepant quality level of histopathological and cytological preparations involved five European countries and exploiting the potential of "virtual slide technology". Concrete issues, techniques and pitfalls, as well as proposed guidelines for processing the tissues are shown in this presentation.
Topics: Histological Techniques; Humans; Preservation, Biological; Tissue Fixation
PubMed: 22732791
DOI: No ID Found -
The Journal of Reproduction and... Oct 2011Freeze-drying (lyophilization) has been proposed as an alternative method for sperm preservation to overcome the disadvantages of the current cryopreservation method... (Review)
Review
Freeze-drying (lyophilization) has been proposed as an alternative method for sperm preservation to overcome the disadvantages of the current cryopreservation method such as the high maintenance cost of frozen stocks, the problems associated with transportation of frozen materials and the potential risk of total loss of the frozen stock. Since freeze-dried spermatozoa after rehydration lose their motility, which is an essential requirement to complete physiological fertilization, a relatively difficult microinsemination technique must be applied to rehydrated spermatozoa. Theoretically, it has been supposed that freeze-dried spermatozoa could maintain their functions and abilities to interact with the oocyte cytoplasm after prolonged storage at refrigerator temperature. However, sufficient yield of transferable blastocysts and production of live offspring derived from freeze-dried sperm samples are still subjects to be challenged and overcome in large domestic species.
Topics: Animals; Cryopreservation; Freeze Drying; Humans; Male; Models, Biological; Reproductive Techniques, Assisted; Semen Preservation; Sperm Injections, Intracytoplasmic; Spermatozoa
PubMed: 22052044
DOI: 10.1262/jrd.11-061o -
Food Research International (Ottawa,... May 2020The objective of this work was to produce solid lipid microparticles using fully hydrogenated anhydrous milk fat (FHAMF) and to evaluate their physical stability during...
The objective of this work was to produce solid lipid microparticles using fully hydrogenated anhydrous milk fat (FHAMF) and to evaluate their physical stability during 90 days of storage at different temperatures. To obtain the lipid microparticles, the FHAMF was sprayed in a double fluid atomizer at 1 bar pressure, in a chilled chamber (2 °C). After atomization, the microparticles were divided into three batches and stored for 90 days at three different temperatures (5, 15 and 25 °C). During storage, samples were periodically removed (7, 15, 30, 60 and 90 days) for evaluation of particle size, melting behavior, morphology, and polymorphic habit. The microparticles presented spherical shaped, with a smooth surface and wide size variation. When stored at 5 °C, the microparticles showed the smaller size and smaller agglomeration, due to the lower liquid fat content in the system, which that makes it difficult the adhesion of one particle to another. The lipid microparticles presented β' crystals immediately after processing and at all temperatures during the storage. This study demonstrated the potential of FHAMF as an appropriate lipid phase for the production of lipid microparticles, and may contribute to further studies on the delivery of active compounds.
Topics: Animals; Dairy Products; Hydrogenation; Lipids; Milk; Nanoparticles; Particle Size; Preservation, Biological; Spray Drying; Temperature
PubMed: 32247456
DOI: 10.1016/j.foodres.2020.109009 -
Journal of Veterinary Science Nov 2023The need for a storage method capable of preserving the intrinsic properties of bones without using toxic substances has always been raised. Supercooling is a relatively...
BACKGROUND
The need for a storage method capable of preserving the intrinsic properties of bones without using toxic substances has always been raised. Supercooling is a relatively recently introduced preservation method that meets this need. Supercooling refers to the phenomenon of liquid in which the temperature drops below its freezing point without solidifying or crystallizing.
OBJECTIVES
The purpose of this study was to identify the preservation efficiency and applicability of the supercooling technique as a cortical bone allograft storage modality.
METHODS
The biomechanical effects of various storage methods, including deep freezing, cryopreservation, lyophilization, glycerol preservation, and supercooling, were evaluated with the three-point banding test, axial compression test, and electron microscopy. Additionally, cortical bone allografts were applied to the radial bone defect in New Zealand White rabbits to determine the biological effects. The degree of bone union was assessed with postoperative clinical signs, radiography, micro-computed tomography, and biomechanical analysis.
RESULTS
The biomechanical properties of cortical bone grafts preserved using glycerol and supercooling method were found to be comparable to those of normal bone while also significantly stronger than deep-frozen, cryopreserved, and lyophilized bone grafts. Preclinical research performed in rabbit radial defect models revealed that supercooled and glycerol-preserved bone allografts exhibited significantly better bone union than other groups.
CONCLUSIONS
Considering the biomechanical and biological superiority, the supercooling technique could be one of the optimal preservation methods for cortical bone allografts. This study will form the basis for a novel application of supercooling as a bone material preservation technique.
Topics: Animals; Rabbits; X-Ray Microtomography; Glycerol; Transplantation, Homologous; Cryopreservation; Cortical Bone; Allografts
PubMed: 37904641
DOI: 10.4142/jvs.23183 -
World Journal of Microbiology &... Nov 2012The aim of the study was to compare the effect of two low-cost, low technology traditional methods for drying starter cultures with standard lyophilisation. Lyophilised...
The aim of the study was to compare the effect of two low-cost, low technology traditional methods for drying starter cultures with standard lyophilisation. Lyophilised yeast cultures and yeast cultures preserved in dry rice cakes and dry plant fibre strands were examined for viable cell counts during 6 months storage at 4 and 25 °C. None of the yeast cultures showed a significant loss in viable cell count during 6 months of storage at 4 °C upon lyophilisation and preservation in dry rice cakes. During storage at 25 °C in the dark, yeast cultures preserved in dry rice cakes, and lyophilised cultures of Saccharomyces cerevisiae and Issatchenkia orientalis showed no significant loss of viable cells up to 4 months of storage. Yeast cultures preserved in dry plant fibre strands had the greatest loss of viable count during the 6 months of storage at 25 °C. Preservation of yeasts cultures in dry rice cakes provided better survival during storage at 4 °C than lyophilisation. The current study demonstrated that traditional methods can be useful and effective for starter culture preservation in small-scale, low-tech applications.
Topics: Desiccation; Freeze Drying; Microbial Viability; Mycology; Pichia; Preservation, Biological; Saccharomyces cerevisiae; Temperature; Time Factors
PubMed: 22806747
DOI: 10.1007/s11274-012-1118-y -
PloS One 2019The preservation of biological samples for an extended time period of days to weeks after initial collection is important for the identification, screening, and...
The preservation of biological samples for an extended time period of days to weeks after initial collection is important for the identification, screening, and characterization of bacterial pathogens. Traditionally, preservation relies on cold-chain infrastructure; however, in many situations this is impractical or not possible. Thus, our goal was to develop alternative bacterial sample preservation and transport media that are effective without refrigeration or external instrumentation. The viability, nucleic acid stability, and protein stability of Bacillus anthracis Sterne 34F2, Francisella novicida U112, Staphylococcus aureus ATCC 43300, and Yersinia pestis KIM D27 (pgm-) was assessed for up to 28 days. Xanthan gum (XG) prepared in PBS with L-cysteine maintained more viable F. novicida U112 cells at elevated temperature (40°C) compared to commercial reagents and buffers. Viability was maintained for all four bacteria in XG with 0.9 mM L-cysteine across a temperature range of 22-40°C. Interestingly, increasing the concentration to 9 mM L-cysteine resulted in the rapid death of S. aureus. This could be advantageous when collecting samples in the built environment where there is the potential for Staphylococcus collection and stabilization rather than other organisms of interest. F. novicida and S. aureus DNA were stable for up to 45 days upon storage at 22°C or 40°C, and direct analysis by real-time qPCR, without DNA extraction, was possible in the XG formulations. XG was not compatible with proteomic analysis via LC-MS/MS due to the high amount of residual Xanthomonas campestris proteins present in XG. Our results demonstrate that polysaccharide-based formulations, specifically XG with L-cysteine, maintain bacterial viability and nucleic acid integrity for an array of both Gram-negative and Gram-positive bacteria across ambient and elevated temperatures.
Topics: Bacteria; Cysteine; Microbial Viability; Polysaccharides; Polysaccharides, Bacterial; Preservation, Biological; Proteomics; Temperature
PubMed: 31490969
DOI: 10.1371/journal.pone.0221831 -
American Journal of Hematology Feb 2010Recent reports suggest that transfusion of old red blood cell (RBC) units (>2 weeks) was associated with increased risks of postoperative complications and higher... (Review)
Review
Recent reports suggest that transfusion of old red blood cell (RBC) units (>2 weeks) was associated with increased risks of postoperative complications and higher mortality rate caught public attention (Yap et al., Ann Thorac Surg 2008; 86:554-559 and Koch et al., 2008; 358:1229-1239). This rekindled the decades old discussion regarding the impact of RBC aging and storage lesions in patient care. The objectives of this review are to provide readers with an overview of the process of banking RBC that may have an impact on its quality, the reported clinical impact of storage lesions, the consequences of transfusing new RBC units only to the nation's blood supply and potential solutions that may improve the feasibility of blood banks to issue new blood units only.
Topics: Blood Banks; Erythrocyte Transfusion; Erythrocytes; Humans; Preservation, Biological
PubMed: 20052749
DOI: 10.1002/ajh.21599 -
Nature Communications Oct 2022Deoxyribonucleic acid (DNA) is the blueprint of life, and cost-effective methods for its long-term storage could have many potential benefits to society. Here we present...
Deoxyribonucleic acid (DNA) is the blueprint of life, and cost-effective methods for its long-term storage could have many potential benefits to society. Here we present the method of in situ cryosilicification of whole blood cells, which allows long-term preservation of DNA. Importantly, our straightforward approach is inexpensive, reliable, and yields cryosilicified samples that fulfill the essential criteria for safe, long-term DNA preservation, namely robustness against external stressors, such as radical oxygen species or ultraviolet radiation, and long-term stability in humid conditions at elevated temperatures. Our approach could enable the room temperature storage of genomic information in book-size format for more than one thousand years (thermally equivalent), costing only 0.5 $/person. Additionally, our demonstration of 3D-printed DNA banking artefacts, could potentially allow 'artificial fossilization'.
Topics: Humans; Ultraviolet Rays; DNA; Blood Preservation; Preservation, Biological; Oxygen
PubMed: 36270991
DOI: 10.1038/s41467-022-33759-y -
Parasites & Vectors Oct 2019Strongyloidiasis is a soil-borne helminthiasis, which, in spite of the up to 370 million people currently estimated to be infected with its causing agent, the nematode...
From the feces to the genome: a guideline for the isolation and preservation of Strongyloides stercoralis in the field for genetic and genomic analysis of individual worms.
Strongyloidiasis is a soil-borne helminthiasis, which, in spite of the up to 370 million people currently estimated to be infected with its causing agent, the nematode Strongyloides stercoralis, is frequently overlooked. Recent molecular taxonomic studies conducted in Southeast Asia and Australia, showed that dogs can carry the same genotypes of S. stercoralis that also infect humans, in addition to a presumably dog-specific Strongyloides species. This suggests a potential for zoonotic transmission of S. stercoralis from dogs to humans. Although natural S. stercoralis infections have not been reported in any host other than humans, non-human primates and dogs, other as yet unidentified animal reservoirs cannot be excluded. Molecular studies also showed that humans carry rather different genotypes of S. stercoralis. As a result, their taxonomic status and the question of whether they differ in their pathogenic potential remains open. It would therefore be very important to obtain molecular genetic/genomic information about S. stercoralis populations from around the world. One way of achieving this (with little additional sampling effort) would be that people encountering S. stercoralis in the process of their diagnostic work preserve some specimens for molecular analysis. Here we provide a guideline for the isolation, preservation, genotyping at the nuclear 18S rDNA and the mitochondrial cox1 loci, and for whole genome sequencing of single S. stercoralis worms. Since in many cases the full analysis is not possible or desired at the place and time where S. stercoralis are found, we emphasize when and how samples can be preserved, stored and shipped for later analysis. We hope this will benefit and encourage researchers conducting field studies or diagnostics to collect and preserve S. stercoralis for molecular genetic/genomic analyses and either analyze them themselves or make them available to others for further analysis.
Topics: Animals; Cyclooxygenase 1; DNA, Helminth; Dog Diseases; Dogs; Feces; Female; Genome; Genotyping Techniques; Humans; Larva; Male; Microspheres; Polymerase Chain Reaction; Preservation, Biological; RNA, Ribosomal, 18S; Soil; Strongyloides stercoralis; Strongyloidiasis; Time Factors; Whole Genome Sequencing
PubMed: 31640777
DOI: 10.1186/s13071-019-3748-5 -
Journal of Anthropological Sciences =... Dec 2017Fossil remains are the only physical evidence of past forms of life which researchers can use to study the evolutionary biology of a species, especially regarding the... (Review)
Review
Fossil remains are the only physical evidence of past forms of life which researchers can use to study the evolutionary biology of a species, especially regarding the human lineage. We review and consider the way in which the conditions surrounding a fossil's discovery and its use for scientific research impacts its long-term preservation. The deterioration of the body starts soon after death, continues in the sediments and only a subsample of the anatomical elements will persist and may finally be unearthed by archeologists. From their recovery onwards, fossil remains are exposed to many sources of further damage: from handling, restoration, measuring to invasive sampling. On the one hand, curators are faced with the inherent challenge of balancing their responsibility to protect fossil specimens with allowing researchers to perform specific analyses or invasive sampling detrimental to the preservation of the fossil. On the other hand, scientists may find their analyses complicated by multiple factors including taphonomy, or restoration techniques (e.g., consolidants, cleaning chemicals). We provide several historical examples illustrating the complex nature of the factors acting on fossil preservation. We discuss concerns about producing and sharing (digital) data from fossils. Finally, we also suggest and support some curatorial practices which maximize the traceability of treatments underwent by a fossil.
Topics: Animals; Fossils; Hominidae; Paleontology; Preservation, Biological
PubMed: 27983518
DOI: 10.4436/JASS.95002