The Effects of Storage Conditions on the Preservation of Enzymatic Activity in Bone

Cosby, Christi N.; Troiano, Nancy; Kacena, Melissa A.

doi:10.1179/his.2008.31.4.169

Cited by 3 publications

(2 citation statements)

References 25 publications

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“…After ethanol storage, variation coefficients for means of phosphatase activity ranged between 28% (day 0), 25% (day 7), 73% (day 14), 38% (day 21), 11% (day 28), and 25% (day 56). Recovery of phosphatase activity after ethanol fixation is in agreement with histochemical methods in which ethanol is used to fix tissues such as bones (Cosby et al 2008) or to carry out post-fixation of preserved tissues (Kacena et al 2004) while maintaining acid phosphatase activity. Secreted acid phosphatase released by the filamentous fungus Aspergillus fumigatus Fresen.…”

Section: Storage Of Ecm As Frozen or Ethanol-fixed Samplessupporting

confidence: 76%

Optimized assay and storage conditions for enzyme activity profiling of ectomycorrhizae

et al. 2011

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The aim of a joint effort by different research teams was to provide an improved procedure for enzyme activity profiling of field-sampled ectomycorrhizae, including recommendations on the best conditions and maximum duration for storage of ectomycorrhizal samples. A more simplified and efficient protocol compared to formerly published procedures was achieved by using manufactured 96-filter plates in combination with a vacuum manifold and by optimizing incubation times. Major improvements were achieved by performing the series of eight enzyme assays with a single series of root samples instead of two series, reducing the time needed for sample preparation, minimizing error-prone steps such as pipetting and morphotyping, and facilitating subsequent DNA analyses due to the reduced sequencing effort. The best preservation of samples proved to be storage in soil at 4-6 °C in the form of undisturbed soil cores containing roots. Enzyme activities were maintained for up to 4 weeks under these conditions. Short-term storage of washed roots and ectomycorrhizal tips overnight in water did not cause substantial changes in enzyme activity profiles. No optimal means for longer-term storage by freezing at -20 °C or storage in 100% ethanol were recommended.

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Section: Storage Of Ecm As Frozen or Ethanol-fixed Samplessupporting

confidence: 76%

Optimized assay and storage conditions for enzyme activity profiling of ectomycorrhizae

et al. 2011

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“…The wet slides were incubated 4 mins with the staining solution and after gently rinsed with dH2O and mounted with Mowiol. Alkaline phosphatase staining to detect osteoblasts was performed as described [25]. Briefly, slides were placed in Tris buffer.…”

Section: Methodsmentioning

confidence: 99%

Blood Clot Formation Does Not Affect Metastasis Formation or Tumor Growth in a Murine Model of Breast Cancer

Rossnagl

Vasel

et al. 2014

PLoS ONE

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Cancer is associated with increased fracture risk, due either to metastasis or associated osteoporosis. After a fracture, blood clots form. Because proteins of the coagulation cascade and activated platelets promote cancer development, a fracture in patients with cancer often raises the question whether it is a pathologic fracture or whether the fracture itself might promote the formation of metastatic lesions. We therefore examined whether blood clot formation results in increased metastasis in a murine model of experimental breast cancer metastasis.For this purpose, a clot was surgically induced in the bone marrow of the left tibia of immundeficient mice. Either one minute prior to or five minutes after clot induction, human cancer cells were introduced in the circulation by intracardiac injection. The number of cancer cells that homed to the intervention site was determined by quantitative real-time PCR and flow cytometry. Metastasis formation and longitudinal growth were evaluated by bioluminescence imaging.The number of cancer cells that homed to the intervention site after 24 hours was similar to the number of cells in the opposite tibia that did not undergo clot induction. This effect was confirmed using two more cancer cell lines. Furthermore, no difference in the number of macroscopic lesions or their growth could be detected. In the control group 72% developed a lesion in the left tibia. In the experimental groups with clot formation 79% and 65% developed lesions in the left tibia (p = ns when comparing each experimental group with the controls). Survival was similar too.In summary, the growth factors accumulating in a clot/hematoma are neither enough to promote cancer cell homing nor support growth in an experimental model of breast cancer bone metastasis. This suggests that blood clot formation, as occurs in traumatic fractures, surgical interventions, and bruises, does not increase the risk of metastasis formation.

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Advanced biomaterials in cell preservation: Hypothermic preservation and cryopreservation

Gao

Tian

et al. 2021

Acta Biomaterialia

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The Effects of Storage Conditions on the Preservation of Enzymatic Activity in Bone

Cited by 3 publications

References 25 publications

Optimized assay and storage conditions for enzyme activity profiling of ectomycorrhizae

Optimized assay and storage conditions for enzyme activity profiling of ectomycorrhizae

Blood Clot Formation Does Not Affect Metastasis Formation or Tumor Growth in a Murine Model of Breast Cancer

Advanced biomaterials in cell preservation: Hypothermic preservation and cryopreservation

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