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1.
PLoS One ; 16(8): e0255363, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34347814

RESUMO

The standard histological processing procedure, which produces excellent staining of sections for most tissues, fails to yield satisfactory results in adult mouse orbits or eyeballs. Here, we show that a protocol using tissue block staining and domestic adhesive tapes resulted in qualified integral serial cryo-sections of whole orbits or eyeballs, and the fine structures were well preserved. The histological processing protocol comprises paraformaldehyde fixation, ethylenediaminetetraacetic acid decalcification, tissue block staining with hematoxylin and eosin, embedding, adhesive tape aided sectioning, and water-soluble mounting. This protocol was proved to be the best in comparison with seven other related existing histological traditional or non-traditional processing methods, according to the staining slice quality. We observed a hundred percent success rate in sectioning, collection, and mounting with this method. The reproducibility tested on qualified section success rates and slice quality scores confirmed that the technique is reliable. The feasibility of the method to detect target molecules in orbits was verified by successful trial tests on block immunostaining and adhesive tape-aided sectioning. Application of this protocol in joints, brains, and so on,-the challenging integral sectioning tissues, also generated high-quality histological staining sections.


Assuntos
Olho/anatomia & histologia , Órbita/anatomia & histologia , Preservação de Tecido/instrumentação , Animais , Criopreservação , Estudos de Viabilidade , Feminino , Camundongos , Microtomia , Coloração e Rotulagem , Fita Cirúrgica , Inclusão do Tecido , Fixação de Tecidos , Preservação de Tecido/métodos
2.
J Pediatr Adolesc Gynecol ; 33(6): 712-714, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32781234

RESUMO

STUDY OBJECTIVE: To present our experience of laparoscopic resection of pediatric benign ovarian teratomas with gonadal preservation, using a homemade glove retrieval bag. DESIGN, SETTING, PARTICIPANTS, INTERVENTIONS, AND MAIN OUTCOME MEASURES: Review of all girls with benign ovarian teratomas who were managed with laparoscopic ovarian-sparing surgery (OSS) at our hospital between January 2013 and December 2018. RESULTS: Eleven patients were included for analysis with a mean age of 6.1 years. Ten patients received elective surgery, whereas 1 patient received emergency surgery because of ovarian torsion. Main indication for OSS was the existence of a dissection plane between tumor margins and healthy ovarian tissue. Postoperative outcome and follow-up were uneventful with a median follow-up of 30.1 months (range; 12-60 months). CONCLUSION: Laparoscopic OSS can be safely performed for these tumors. Laparoscopic magnification with energy devices are excellent tools in such procedures. The homemade glove bag can be used to retrieve the tumor effectively in countries with limited resources.


Assuntos
Laparoscopia/métodos , Neoplasias Ovarianas/cirurgia , Ovário/cirurgia , Teratoma/cirurgia , Preservação de Tecido/instrumentação , Adolescente , Criança , Pré-Escolar , Desenho de Equipamento , Feminino , Humanos , Lactente
3.
Cell Tissue Bank ; 21(1): 89-97, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31838727

RESUMO

Precision tissue diagnostics rely on high quality input specimens so that assay results are not affected by artifact, but advances in collection and processing of tissue specimens have lagged behind innovations in diagnostic assay development. Therefore, we have designed and evaluated a novel surgical tissue collection device that maintains and monitors sample temperature and motion throughout transport so that the major preanalytical variable of tissue temperature can be controlled and measured. This device, in combination with an improved cold-hot tissue fixation protocol affords optimal biomarker preservation in less overall time, thereby simultaneously improving diagnostic quality and turnaround time. We collected 50 primary and metastatic liver tumors using a novel transport device. Tissue was fixed using a rapid cold-hot fixation protocol and immunohistochemical assays were used to assess the performance of the device, in comparison to control tissue preserved using standard clinical fixation protocol. Two pathologists evaluated the IHC studies in a blinded fashion to determine the immunophenotype of each tumor. The observed IHC staining intensities and the clinical impressions of the immunophenotypes did not differ between tissue collected with the novel device and control tissue, while improvements in processing time were achieved. The novel cold transport device and rapid fixation protocol can be successfully and safely combined and used to monitor specimen conditions, thus preserving the diagnostic utility of specimens and improving the overall turn-around time of the diagnostic process.


Assuntos
Biomarcadores Tumorais/análise , Biópsia/instrumentação , Neoplasias/patologia , Fixação de Tecidos/instrumentação , Preservação de Tecido/instrumentação , Biópsia/economia , Temperatura Baixa , Desenho de Equipamento , Humanos , Imuno-Histoquímica , Temperatura , Fatores de Tempo , Fixação de Tecidos/economia , Preservação de Tecido/economia
4.
Medicine (Baltimore) ; 97(50): e13175, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30557967

RESUMO

RATIONALE: In this report, a combination of socket-shield technique (SST) and platelet-rich fibrin (PRF) technique was used for immediate implant placement on a fractured central incisor. During the follow-up visit, cone beam computed tomography (CBCT) and clinical observation were used to evaluate the preservation outcome of peri-implant bone and gingiva. PATIENT CONCERNS: The patient was a 28-year-old healthy female patient who desired her fractured 21 to be replaced with an implant-supported single crown; the fractured 21 comprised a post-core crown with insufficient residual bone at the labial site. DIAGNOSIS: The root of 21 exhibited a complex root fracture; the labial portion of the alveolar ridge was thin (<1 mm) and partial ankylosis of the residual root was observed. INTERVENTIONS: Modified SST was applied to the labial portion of the residual root. The implant was placed immediately at the lingual site of the retained socket-shield root fragment; PRF was the placed in the gap between the root fragment and the implant. Final prosthodontic treatment was performed at 24 weeks after implant placement. OUTCOMES: Clinical examination and CBCT scanning at various follow-up visits time showed that the periodontal tissue was well- preserved. At 6 months after surgery, the average horizontal and vertical peri-implant bone resorption was 0.4 mm; a follow-up visit at 18 months post-loading indicated that peri-implant tissue was well preserved by the shield-technique and no significant peri-implant tissue resorption was displayed. LESSON SUBSECTIONS: In cases of anterior teeth with intact but insufficient residual alveolar ridge, the SST with PRF may be effective for preservation and maintenance of stable peri-implant tissue.


Assuntos
Incisivo/efeitos dos fármacos , Incisivo/cirurgia , Fibrina Rica em Plaquetas/efeitos dos fármacos , Adulto , Coroas , Feminino , Fraturas Ósseas/diagnóstico por imagem , Fraturas Ósseas/tratamento farmacológico , Fraturas Ósseas/cirurgia , Humanos , Preservação de Tecido/instrumentação , Preservação de Tecido/métodos , Raiz Dentária/diagnóstico por imagem , Raiz Dentária/efeitos dos fármacos , Raiz Dentária/cirurgia
5.
Forensic Sci Int Genet ; 36: 124-129, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29990824

RESUMO

Short tandem repeats (STR) are currently the gold standard in human identification for forensic casework purposes, and successful STR typing is dependent on sufficient quantity and quality DNA. In the aftermath of a mass disaster and some forensic cases, human remains are recovered for identification in various stages of decomposition, and ideally these remains are transported to a refrigerated facility in order to halt the decomposition process and preserve the integrity of DNA within the tissue. However, in situations where refrigeration is not available (e.g., after a mass disaster or in rural forensic casework), remains continue to be exposed to environmental insults after collection, causing further DNA damage and degradation. Therefore, successful STR typing is dependent on the time of collection and preservation of the DNA sample. This study aims to test two simple in-field collection and preservation methods for decomposing human tissues that are subsequently stored at room temperature for up to six months either in a tissue preservative solution (modified TENT buffer) or on an FTA® Elute Card. In addition, these collection and preservation methods were tested for their ability to facilitate more direct and faster processing of DNA from preserved tissues or DNA leached into the surrounding TENT preservative solution for STR typing. Pre-PCR methods tested in this study include a quick lysis of FTA® Elute Cards, silica-based purification (QIAquick®), enzyme-based extractions (PDQeX), and simple dilution of liquid preservative. The traditional DNA analysis pipeline, which includes DNA extraction and quantification, will be compared to an alternate direct PCR method, thereby allowing the elimination of these two time-consuming and costly steps. The results indicate that modified TENT preservative and FTA® Elute Cards both preserved DNA from relatively fresh tissue for up to six months at room temperature. However, mostly partial profiles were produced from decomposed tissues (day 6 - day 14 in this study) when stored for up to six months compared to when tissues were processed immediately following collection. Overall, the modified TENT preservative produced higher DNA concentrations and more successful STR results than FTA® Elute Cards. In addition, a rapid DNA extraction platform (PDQeX) generated the most successful STR typing results from the decomposed tissues stored in TENT for up to six months at room temperature. The direct PCR method used in this study generated comparable STR results to the traditional DNA analysis approach, warranting further investigation of direct PCR methods for forensic casework type samples.


Assuntos
Restos Mortais , Impressões Digitais de DNA , Repetições de Microssatélites , Manejo de Espécimes , Preservação de Tecido , Genética Forense/instrumentação , Genética Forense/métodos , Humanos , Soluções para Preservação de Órgãos , Reação em Cadeia da Polimerase , Mudanças Depois da Morte , Manejo de Espécimes/instrumentação , Manejo de Espécimes/métodos , Preservação de Tecido/instrumentação , Preservação de Tecido/métodos
6.
Transplant Proc ; 50(2): 416-417, 2018 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-29579817

RESUMO

The bags used in the transport of organs and tissues must be sterile, nontoxic, pyrogen free, and must serve as a barrier throughout their useful life. The goal of this study was to show the sterility, safety, and functionality of the bags subjected to irradiation, through validated procedures and techniques. The selected sterilization method was the use of gamma radiation. The sterilization dose was determined based on validated standards for the sterilization of medical products, ISO 11137-2: 2013 and ISO/TS 13004: 2013, using the Verification Dose Maximum method on samples belonging to 3 manufacturing lots. The ISO 10993-5: 2009 standard was used in the cytotoxicity tests, by means of extracts test and quantitative technique of MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide. The tests to determine the expiration date of the kit were performed by ASTM F1980, accelerated aging, and ASTM D3078 to evaluate hermeticity. The irradiation dose validated to reach the required sterility safety level was 22.5 kGy. The constituent materials and the sterilization method do not generated cellular toxicity, and the product was not modified during the simulated time of 5 years. Sterilization by irradiation is a method that leaves no residue, does not harm the properties of the material because it is conducted in cold, and as the sterilizing agent, the energy absorbed by the product is highly penetrating and can be treated in its final packaging, with no risk of postcontamination. It is for this reason that it is prioritized over other methods of sterilization.


Assuntos
Preservação de Órgãos/instrumentação , Embalagem de Produtos/métodos , Esterilização/métodos , Preservação de Tecido/instrumentação , Raios gama , Humanos
7.
Biopreserv Biobank ; 15(5): 417-421, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28745913

RESUMO

Tears covering the ocular surface are important biofluids containing thousands of molecules, including proteins, lipids, metabolites, nucleic acids, and electrolytes. Tears are valuable resources for biomarker research of ocular and even systemic diseases. For application in biomarker studies, tear samples should ideally be stored using a simple, lowcost, and efficient method along with the patient's medical records. For this purpose, we developed a novel Schirmer's strip-based dry method that allows for storage of tear samples in vacuum bags at room temperature. Using this method, tear protein patterns can also be preserved. Liquid chromatography-mass spectrometry/mass spectrometry analysis of proteins recovered by the dry method and traditional wet method showed no significant difference. Some tissue/organ-enriched proteins were identified in tear samples, thus tears might be a good window for monitoring changes of these tissues or organs. This dry method facilitates sample transportation and enables the storage of tear samples on a large scale, increasing the availability of samples for studying disease biomarkers in tears.


Assuntos
Dessecação/métodos , Lágrimas/metabolismo , Preservação de Tecido/métodos , Cromatografia Líquida , Proteínas do Olho/análise , Humanos , Espectrometria de Massas em Tandem , Preservação de Tecido/instrumentação
8.
J Vis Exp ; (122)2017 04 06.
Artigo em Inglês | MEDLINE | ID: mdl-28448056

RESUMO

Studies on skeletal muscle physiology face the technical challenge of appropriately processing the specimens to obtain sections with clearly visible cytoplasmic compartments. Another hurdle is the tight apposition of myofibers to the surrounding tissues. Because the process of tissue fixation and paraffin embedding leads to the shrinkage of muscle fibers, freezing is an optimal means of hardening muscle tissue for sectioning. However, a commonly encountered issue, the formation of ice crystals, occurs during the preparation of frozen sections because of the high water content of muscle. The protocol presented here first describes a simple and efficient method for properly freezing muscle tissues by immersing them in liquid nitrogen. The problem with using liquid nitrogen alone is that it causes the formation of a nitrogen gas barrier next to the tissue, which acts as an insulator and inhibits the cooling of the tissues. To avoid this "vapor blanket" effect, a new cryovial was designed to increase the speed of liquid flow around the tissue surface. This was achieved by punching a total of 14 inlet holes in the wall of the vial. According to bubble dynamics, a higher rate of liquid flow results in smaller bubbles and fewer chances to form a gas barrier. When liquid nitrogen flows into the cryovial through the inlet holes, the flow velocity around the tissue is fast enough to eliminate the gas barrier. Compared to the method of freezing muscle tissues using pre-chilled isopentane, this protocol is simpler and more efficient and can be used to freeze muscle in a throughput manner. Furthermore, this method is optimal for institutions that do not have access to isopentane, which is extremely flammable at room temperature.


Assuntos
Artefatos , Congelamento , Secções Congeladas , Músculo Esquelético , Fixação de Tecidos/métodos , Preservação de Tecido/instrumentação , Preservação de Tecido/métodos , Desenho de Equipamento
9.
Fertil Steril ; 106(6): 1348-1355, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27490043

RESUMO

OBJECTIVE: To evaluate whether is possible to vitrify oocytes in an aseptic (hermetically closed) fashion and maintain clinical results comparable with those of fresh oocytes. DESIGN: Prospective, observational, cohort, noninferiority trial. SETTING: Private in vitro fertilization center. PATIENT(S): One hundred eighty-four recipients of donated vitrified oocytes. INTERVENTION(S): Closed system vitrification. MAIN OUTCOME MEASURE(S): Pregnancy rate per cycle and clinical pregnancy rate per cycle. RESULT(S): No statistically significant differences were observed between two groups regarding the pregnancy rate per cycle (63.1% vs. 60.9%) or the clinical pregnancy rate per cycle (55.4% vs. 58.7%). Biochemical pregnancy rate was statistically significantly higher in the fresh group (7.6% vs. 2.2%). The mean number of embryos transferred was similar (2.0 ± 0.0 vs. 1.97 ± 0.3). Concerning embryologic data, there were no statistically significant differences regarding the fertilization, cleavage, top quality day-3 embryo, or blastocyst rates, whereas the top quality blastocyst rate on day 5 was statistically significantly higher in the fresh oocyte group (31.7% vs. 26.1%). CONCLUSION(S): Aseptically (in a closed system) vitrified oocytes show similar clinical efficiency compared with their sibling fresh oocytes.


Assuntos
Assepsia/métodos , Criopreservação/métodos , Infertilidade/terapia , Doação de Oócitos , Preservação de Tecido/métodos , Adulto , Assepsia/instrumentação , Criopreservação/instrumentação , Transferência Embrionária , Feminino , Fertilidade , Fertilização in vitro , Humanos , Infertilidade/diagnóstico , Infertilidade/fisiopatologia , Nascido Vivo , Masculino , Pessoa de Meia-Idade , Gravidez , Taxa de Gravidez , Estudos Prospectivos , Preservação de Tecido/instrumentação , Resultado do Tratamento , Vitrificação
10.
Ann Plast Surg ; 76(3): 355-60, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26808757

RESUMO

BACKGROUND: Traumatic amputation is the second leading cause of limb loss in the United States. The preferred treatment is salvage and replantation of the amputated limb, whenever possible, and allotransplantation is a novel procedure whereby healthy limbs are procured from deceased organ donors and transplanted into the amputee recipient. A major restriction for both procedures is the irrecoverable muscle damage occurring due to ischemia. We investigated the feasibility of using a novel lightweight, mobile perfusion device specifically designed to perfuse amputated porcine limbs with an acellular perfusion solution to delay ischemic muscle damage prior to transplantation or replantation. METHODS: Bilateral hind limbs of Yorkshire pigs were amputated; one of the limbs was preserved by perfusion in the mobile perfusion device, and the other by storage in ice slurry for 12 hours. RESULTS: Five sets of bilateral limbs were preserved as described previously. A defined pressure of 30 mm Hg was reliably maintained in the arterial system without loss of flow. Comparison of the perfusate composition before and after limb passage revealed significant differences. Muscle biopsies showed a consistent progression of clusters of hypoxic cells in the control limbs with time. Similar changes could not be observed in the perfused tissue. CONCLUSIONS: We have designed and built a small, mobile perfusion device that is operational and that more closely mimics the normal physiological environment when compared with the current standard of preservation in ice slurry. This project may have far-reaching implications for the treatment of limb loss through replantation and transplantation.


Assuntos
Amputação Traumática/cirurgia , Complicações Pós-Operatórias/prevenção & controle , Traumatismo por Reperfusão/prevenção & controle , Reimplante , Preservação de Tecido/instrumentação , Amputação Traumática/patologia , Animais , Estudos de Viabilidade , Feminino , Salvamento de Membro , Complicações Pós-Operatórias/patologia , Distribuição Aleatória , Traumatismo por Reperfusão/etiologia , Traumatismo por Reperfusão/patologia , Suínos , Preservação de Tecido/métodos , Resultado do Tratamento , Alotransplante de Tecidos Compostos Vascularizados
11.
Kyobu Geka ; 68(11): 903-6, 2015 Nov.
Artigo em Japonês | MEDLINE | ID: mdl-26469255

RESUMO

From August 2003 to June 2013, 9 patients with aortic valve endocarditis underwent aortic root replacement using homografts which were harvested and preserved in our institute. The median patient age was 62 years (range 46~84) and 5 patients were men. Four cases were prosthetic valve infections. The in-hospital mortality was 0%. In 8 of 9 cases were evaluated on midterm outcomes. At a median of 52 months (range 19~156), overall survival was 100%, freedom from cardiovascular events was 87.5%. The peak aortic pressure gradient was 9.04 ± 4.2 mmHg. Aortic regurgitation was less than 2 of 4 in all cases.


Assuntos
Valva Aórtica/transplante , Endocardite Bacteriana/cirurgia , Cardiopatias Congênitas/cirurgia , Doenças das Valvas Cardíacas/cirurgia , Idoso , Valva Aórtica/fisiopatologia , Valva Aórtica/cirurgia , Doença da Válvula Aórtica Bicúspide , Feminino , Cardiopatias Congênitas/fisiopatologia , Doenças das Valvas Cardíacas/fisiopatologia , Humanos , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/cirurgia , Preservação de Tecido/instrumentação , Transplante Homólogo , Resultado do Tratamento
12.
Recent Results Cancer Res ; 199: 15-26, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25636425

RESUMO

We describe five validation trials of new vacuum sealing technologies that change the approach to the preanalytic "front end" of specimen transport, handling, and processing and illustrate their adaptation and integration into existing Lean laboratory operations with reduction in formalin use and personnel exposure to this toxic and potentially carcinogenic fixative. These trials provide histologic assessment by numerous pathologists of tissues processed in this new paradigm and define the financial advantages of applying this technology to the postanalytic or "back end" process of tissue storage. We conclude that the TisssueSAFE and SealSAFE vacuum sealing systems are both promising technologies for preserving fresh human specimens that can promote a safer environment by markedly reducing formalin use in operating room theaters and can minimize formalin use by laboratories.


Assuntos
Manejo de Espécimes , Preservação de Tecido , Vácuo , Formaldeído , Técnicas Histológicas , Humanos , Manejo de Espécimes/instrumentação , Manejo de Espécimes/métodos , Manejo de Espécimes/normas , Temperatura , Fatores de Tempo , Preservação de Tecido/instrumentação , Preservação de Tecido/métodos , Preservação de Tecido/normas , Coleta de Tecidos e Órgãos/métodos , Coleta de Tecidos e Órgãos/normas , Meios de Transporte/métodos
13.
Recent Results Cancer Res ; 199: 119-33, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25636435

RESUMO

In anatomic pathology, the current state encompassing the pre-analytic processes of tissue collection, handling, examination, preparation, processing, and storage are largely uncontrolled, inconsistently performed, and/or not standardized according to the sound scientific data. Pre-analytic defects result in nearly three-quarters of the problems in laboratory diagnostics. This is evident in quality surveys from well-respected institutions that document high miss rates in the required basics of information related to patient and tissue identity, let alone parameters documenting quality aspects related to the surgical specimen and its preservation. This talk will describe the historical approach to tissue processing and identify gaps from worldwide observations in current laboratory practices. It will also offer potential methodological and technological solutions and process improvements that laboratories may consider in serving the ultimate users of pathology information: the clinician and the patient. It illustrates the need for scientifically validated specimen guidelines and a performance based, standardized and documented "chain of custody" of the pre-analytical steps from the patient's body through fixation. For thought leaders and professional standard setters, opportunities for optimizing molecular studies exist in specimen collection, transfer, grossing, fixation, and decalcification protocols. In this evolving era of molecular profiling and personalized therapeutic decision-making, a well-reasoned and coordinated focus on pre-analytic processes that optimizes specimens for subsequent testing will result in: Improved specimen quality for molecular testing Improved accuracy of diagnostic and molecular test results Reduced Turnaroundtimes for same-day diagnosis Enhanced satisfaction of clinicians and patients.


Assuntos
Serviços de Laboratório Clínico/tendências , Padrões de Prática Médica/tendências , Manejo de Espécimes , Serviços de Laboratório Clínico/normas , Técnica de Descalcificação/instrumentação , Técnica de Descalcificação/métodos , Humanos , Laboratórios/tendências , Microtomia/instrumentação , Microtomia/normas , Microtomia/tendências , Padrões de Prática Médica/normas , Manejo de Espécimes/instrumentação , Manejo de Espécimes/normas , Manejo de Espécimes/tendências , Preservação de Tecido/instrumentação , Preservação de Tecido/normas , Preservação de Tecido/tendências , Coleta de Tecidos e Órgãos/instrumentação , Coleta de Tecidos e Órgãos/normas , Coleta de Tecidos e Órgãos/tendências , Meios de Transporte/instrumentação , Meios de Transporte/normas , Vácuo
14.
Clin Biochem ; 47(4-5): 280-7, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24565988

RESUMO

Biospecimen science has recognized the importance of tissue quality for accurate molecular and biomarker analysis and efforts are made to standardize tissue procurement, processing and storage conditions of tissue samples. At the same time the field has emphasized the lack of standardization of processes between different laboratories, the variability inherent in the analytical phase and the lack of control over the pre-analytical phase of tissue processing. The problem extends back into tissue samples in biorepositories, which are often decades old and where documentation about tissue processing might not be available. This review highlights pre-analytical variations in tissue handling, processing, fixation and storage and emphasizes the effects of these variables on nucleic acids and proteins in harvested tissue. Finally current tools for quality control regarding molecular or biomarker analysis are summarized and discussed.


Assuntos
Artefatos , Bancos de Espécimes Biológicos/organização & administração , Manejo de Espécimes/normas , Preservação de Tecido/métodos , Biomarcadores/análise , Fixadores/química , Guias como Assunto , Humanos , Inclusão em Parafina , Estabilidade Proteica , Controle de Qualidade , Estabilidade de RNA , Manejo de Espécimes/métodos , Fatores de Tempo , Preservação de Tecido/instrumentação
15.
Clin Biochem ; 47(4-5): 267-73, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24362270

RESUMO

UNLABELLED: Frozen biospecimens are crucial for translational research and contain well-preserved nucleic acids and protein. However, the risks of freezer failure as well as space, cost, and environmental concerns of frozen biospecimens are substantial. OBJECTIVE: The purpose of the study was to review the current status of room temperature biospecimen storage. METHODS: We searched Pubmed and vendor websites to identify relevant information. RESULTS: Formalin-fixed paraffin embedded (FFPE) tissues have great value but their use is limited by cross-linking and fragmentation of nucleic acids, as well as loss of enzymatic activity. Stabilization solutions can now robustly preserve fresh tissue for up to 7days at room temperature. For longer term storage, commercial vendors of chemical matrices claim real time stability of nucleic acids of over 2 years and their accelerated aging studies to date suggest stability for 12years for RNA and 60years for DNA. However, anatomic pathology biorepositories store mostly frozen tissue rather than nucleic acids. Small quantities of tissue can be directly placed on some chemical matrices to stabilize DNA, however RNA and proteins are not preserved. Current lyophilization approaches can preserve histomorphology, DNA, RNA, and proteins though RNA shows moderate degradation after 1-2years. Formalin-free fixatives show improved but varying abilities to preserve nucleic acids and face validation as well as cost barriers in replacing FFPE specimens. The paraffin embedding process can degrade RNA. CONCLUSION: Development of robust long-term room temperature biospecimen tissue storage technology can potentially reduce costs for the biomedical community in the face of growing targeted therapy needs and decreasing budgets.


Assuntos
Bancos de Espécimes Biológicos/organização & administração , Pesquisa Biomédica/organização & administração , Manejo de Espécimes/normas , Preservação de Tecido/métodos , Fixadores/química , Liofilização , Humanos , Inclusão em Parafina , Estabilidade Proteica , Controle de Qualidade , Estabilidade de RNA , Manejo de Espécimes/economia , Manejo de Espécimes/métodos , Temperatura , Fatores de Tempo , Preservação de Tecido/economia , Preservação de Tecido/instrumentação
16.
J Matern Fetal Neonatal Med ; 25(6): 587-94, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-21819308

RESUMO

OBJECTIVE: Umbilical cord blood gas analysis has a significant and growing role in early neonatal assessment. Factors often delay analysis of cord blood allowing values to change. Consequently, this study evaluates the impact of time, temperature and method of storage on umbilical blood gas and lactate analyses. METHODS: Umbilical cord segments from 80 singleton deliveries were randomized to: cords at room temperature (CR), cords stored on ice (CI), syringes at room temperature (SR) or syringes stored on ice (SI). Analysis occurred every 15 minutes for one-hour. Mixed model analysis of variance allowing for repeated measures was utilized. RESULTS: Cord arterial pH deteriorated in CR, CI, and SI within 15 minutes (p ≤ 0.001), with SR stable until 60 minutes (p = 0.002). Arterial pCO(2) remained stable in SR and CI, increased in SI (p = 0.002; 45 minutes) and decreased in CR (p < 0.001; 45 minutes). Arterial base excess deteriorated in CR and SI (p ≤ 0.009; 15 minutes), SR (p < 0.001; 30 minutes), and CI (p < 0.001; 45 minutes). Arterial lactate levels increased within 15 minutes in all groups (p < 0.001). CONCLUSIONS: Cord blood gas values change rapidly after delivery. Smallest changes were seen in SR group. Data suggest that analyses should be conducted as soon as possible after delivery.


Assuntos
Sangue Fetal/química , Gases/sangue , Ácido Láctico/sangue , Temperatura , Preservação de Tecido/instrumentação , Preservação de Tecido/métodos , Índice de Apgar , Asfixia Neonatal/sangue , Asfixia Neonatal/diagnóstico , Análise Química do Sangue/instrumentação , Análise Química do Sangue/métodos , Gasometria/métodos , Parto Obstétrico/métodos , Equipamentos e Provisões , Feminino , Gases/análise , Idade Gestacional , Humanos , Recém-Nascido , Ácido Láctico/análise , Triagem Neonatal/métodos , Gravidez , Fatores de Tempo
17.
Tissue Eng Part C Methods ; 15(3): 345-53, 2009 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-19196126

RESUMO

A bioartificial liver (BAL) assist system employing a three-dimensional (3D) bioreactor has been studied as a temporary support in liver failure. In the present study, a novel preservation method of primary cultured porcine hepatocytes in monolayer and 3D culture systems was studied. Epigallocatechin-3-gallate (EGCG), which has recently been found to have various bioactivities, was selected as a key compound for hepatocyte preservation. Hepatocytes isolated from porcine liver using the collagenase perfusion method were pre-cultured for 6 days, preserved at room temperature in the presence of EGCG at various concentrations for 4 days, and post-cultured in normal medium for another 6 days. In the monolayer culture, only albumin production rate was fully recovered after preservation when EGCG concentration was high (0.25mg/mL). In contrast, albumin production and ammonium metabolism in the 3D bioreactor under the same condition recovered to 72+/-16% and 98+/-32%, respectively, of levels before preservation. These results indicate that hepatocytes can be preserved in the presence of 0.25mg/mL of EGCG at room temperature, especially in a 3D culture system, which is promising technology for BAL preparation.


Assuntos
Reatores Biológicos , Catequina/análogos & derivados , Técnicas de Cultura de Células/instrumentação , Hepatócitos/fisiologia , Fígado Artificial , Engenharia Tecidual/instrumentação , Preservação de Tecido/instrumentação , Animais , Catequina/farmacologia , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Desenho de Equipamento , Análise de Falha de Equipamento , Hepatócitos/citologia , Hepatócitos/efeitos dos fármacos , Suínos , Porco Miniatura , Temperatura , Preservação de Tecido/métodos
19.
Dent Traumatol ; 24(4): 422-9, 2008 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-18721341

RESUMO

Healing following replantation of avulsed teeth is dependent upon short unphysiologic periods during the extraalveolar phase. A commercially available tooth rescue box was developed and distributed at schools in Hessen, a state in Germany. Aim of the study was to evaluate the availability times of rescue boxes and the storage periods of rescued teeth within the boxes. Two thousand one hundred tooth rescue boxes together with a questionnaire were distributed predominantly at schools. In case of usage of a box, the questionnaire should be filled out by patients and dentists and sent back for evaluation. One hundred seventy-two (8.2%) questionnaires were sent back. Eighteen questionnaires were incomplete. In the remaining 154 tooth rescue boxes, a total of 201 avulsed teeth and tooth crown fragments were rescued. When accidents occurred near a stored rescue box, the availability time was short (median: 5 min). It was significantly longer (median: 35 min) when the location of the accident was distant to a stored box. Storage of avulsed teeth in the tooth rescue box was longer (median: 2 h) than storage of fractured crown fragments (median: 1 h). Lay people (teachers, pupils) used the rescue boxes correctly without professional help or even advice through telephone. The usage of the tooth rescue box seemed to be self-explanatory and plausible to lay persons, very short availability times resulted when accidents occurred near stored boxes. Thus, an excellent healing prognosis can be anticipated after replantation. The storage periods of avulsed teeth before the commencement of treatment exceed by far the periods that are acceptable for alternative but unphysiologic media (saline, saliva, milk). It is concluded that tooth rescue boxes should be distributed at locations prone to tooth traumas (schools, kindergartens, sporting facilities, public pools) to enhance the prognosis of avulsed teeth. Emergency units (hospitals, ambulances) should be equipped with tooth rescue boxes as well as every dentist. Tooth rescue boxes are recommended for families with children.


Assuntos
Soluções para Preservação de Órgãos , Preservação de Tecido/instrumentação , Avulsão Dentária/terapia , Análise de Variância , Distribuição de Qui-Quadrado , Criança , Humanos , Inquéritos e Questionários , Fatores de Tempo
20.
Curr Protoc Cell Biol ; Chapter 4: Unit4.17, 2007 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-18228517

RESUMO

The surface of metazoan cells is a landscape not clearly visualized by light microscopy. Many cells elaborate protrusive structures such as microvilli, filopodia, lamellipodia, and surface ruffles that play important roles in the interaction between the cell and its environment. The high resolution of scanning electron microscopy makes it an ideal technique for studies of the cell surface; however, preservation of fine surface structure can be problematic. Here we highlight the critical factors in sample preparation to ensure optimal high-resolution imaging of the surface of mammalian cells and tissues.


Assuntos
Extensões da Superfície Celular/ultraestrutura , Microscopia Eletrônica de Varredura/instrumentação , Microscopia Eletrônica de Varredura/métodos , Preservação de Tecido/métodos , Animais , Comunicação Celular , Células Epiteliais/ultraestrutura , Microscopia Eletrônica de Varredura/tendências , Microvilosidades/ultraestrutura , Pseudópodes , Preservação de Tecido/instrumentação , Preservação de Tecido/tendências
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