Digitum Colección:
http://hdl.handle.net/10201/123207
2024-03-19T07:16:24ZStorage of blood clots for histological analysis: how long is too long in saline and paraformaldehyde?
http://hdl.handle.net/10201/125427
Título: Storage of blood clots for histological analysis: how long is too long in saline and paraformaldehyde?
Autor/es principal/es: Douglas, Andrew; Fitzgerald, Seán; Pandit, Abhay; Doyle, Karen M.
Resumen: To investigate the composition of blood clots
following mechanical thrombectomy, it is essential to
ensure optimum storage for highest quality histological
and immunofluorescence analysis. We investigated for
how long clots can be stored in paraformaldehyde (PFA),
saline and heparinised saline before the tissue integrity is
compromised.
Whole blood and fibrin-rich clot analogues were
made under dynamic flow conditions. Clots were stored
in 4% PFA, saline or heparinised saline for timepoints
ranging from 1 hour to two months. Five μm sections
were stained with Martius Scarlet Blue to visualise red
blood cells (RBCs), white blood cells (WBCs) and
fibrin. Semi-quantitative analysis of the integrity of clot
components used a scoring system (0: Poor; 1: Sub-par;
2: High). Quantitative analysis used Orbit Image
Analysis software. Autofluorescence was assessed using
a relative scale.
Clots stored in PFA for up to two months were
qualitatively similar to those stored for all shorter
periods (median score: 2 per component). Clots stored in
saline/heparinised saline for one week showed
degradation of RBCs and WBCs, but fibrin remained
intact (median score: 1, 1, 2 respectively). Degradation
of the samples stored in saline/heparinised saline made
accurate quantification using Image Analysis software
difficult from 24h.
Samples stored in PFA for up to two weeks showed
an edging autofluorescence effect, which became more
evident with prolonged storage.
For optimum histology, ideally clots should not be
stored in saline before fixation and should ideally be
stored in formalin for less than one month to minimise
the impact of autofluorescence on immunofluorescence2020-01-01T00:00:00ZSilencing of synaptotagmin 7 regulates osteosarcoma cell proliferation, apoptosis, and migration
http://hdl.handle.net/10201/125426
Título: Silencing of synaptotagmin 7 regulates osteosarcoma cell proliferation, apoptosis, and migration
Autor/es principal/es: Wu, Zhiqiang; Sun, Zhengwang; Huang, Rui; Zang, Ding; Wang, Chunmeng; Yan, Xu; Yan, Wangjun
Resumen: Background. Synaptotagmin 7 (SYT7) is a
component of the synaptotagmin family, which is
essential in many physiological and pathological
processes. In this study, we aimed to investigate the role
of SYT7 in osteosarcoma.
Methods. We defined the expression levels of SYT7
in osteosarcoma tissues and para-sarcoma tissues by
immunohistochemistry and analyzed the possible
correlation between SYT7 expression and pathological
characteristics via Mann-Whitney U analysis and
Spearman correlation analysis. The effects of SYT7
silencing in vitro on cell growth were assessed by MTT
assay. Cell cycle and cell apoptosis were assessed by
flow cytometry analysis. Wound healing assay and
transwell assay were applied to assess the migration and
invasion capacity.
Results. The results showed that the expression
levels of SYT7 were upregulated in osteosarcoma tissues
compared with para-sarcoma tissues and positively
correlated with the pathological characteristics of
osteosarcoma. Functional experiments demonstrated that
SYT7 silencing significantly inhibited cell proliferation
and colony formation capacity (P<0.001), induced cell
cycle arrest which increased the proportion of G2 phase
and decreased the proportion of S phase, enhanced cell
apoptosis (P<0.01), and limited the capacity of migration
and invasion (P<0.01), compared with shCtrl group.
Conclusion. The results indicated that SYT7 plays a
crucial role in the development of osteosarcoma. SYT7
can be applied as a new diagnostic and therapeutic target
in osteosarcoma.2020-01-01T00:00:00ZAdipose-derived stem cells and adipose-derived stem cell-conditioned medium modulate in situ imbalance between collagen I- and collagen V-mediated IL-17 immune response recovering bleomycin pulmonary fibrosis
http://hdl.handle.net/10201/125425
Título: Adipose-derived stem cells and adipose-derived stem cell-conditioned medium modulate in situ imbalance between collagen I- and collagen V-mediated IL-17 immune response recovering bleomycin pulmonary fibrosis
Autor/es principal/es: Gonçalves Felix, Renato; Carvalho Bovolato, Ana Livia; Cotrim, Ondina Silvia; Leão, Patricia dos Santos; Batah, Sabrina Setembre; Golim, Márjorie de Assis; Velosa, Ana Paula P.; Teodoro, Walcy; Martins, Vanessa; Ferreira Cruz, Fernanda; Deffune, Elenice; Fabro, Alexandre Todorovic; Capelozzi, Vera Luiza
Resumen: The immunogenic collagen V (Col V) and the
proinflammatory cytokine interleukin (IL)-17 have been
implicated in the pathogenesis of multiple autoimmune
diseases. Col V is also up-regulated during adipogenesis
and can stimulate adipocyte differentiation in vitro.
Conditioned medium (CM) generated from adipose-
derived mesenchymal stem cells (MSCs) reduces
bleomycin (BLM)-induced lung injury in rats, suggesting
a crucial role in situ of immunomodulatory factors
secreted by MSCs in these beneficial effects. In the
present work, we investigated this hypothesis, analyzing
levels of plasma inflammatory mediators and
inflammatory and fibrotic mediators in the lung tissue of
BLM-injured rats after treatment with MSCs and CM.
Pulmonary fibrosis was intratracheally induced by BLM.
After 10 days, BLM animals were further randomized
into subgroups receiving saline, MSCs, or CM
intravenously. On days 14 and 21, the animals were
euthanized, and the lungs were examined through protein
expression of nitric oxide synthase (NOS), IL-17,
transforming growth factor-β (TGF-β), vascular
endothelial growth factor, endothelin-1, and the
immunogenic Col V through histological quantitative
evaluation and plasma levels of fibrinogen, Von
Willebrand factor, and platelet-derived growth factor
(PDGF). Rats that had been injected with MSCs and CM
showed a significant increase in weight and significant
improvements at 14 and 21 days after intravenous
injection at both time points of analysis of plasma
fibrinogen, PDGF, and Von Willebrand factor and NOS-2
expression, supporting an early anti-inflammatory action,
thus reducing TGF-β and collagen I fibers. In contrast,
intravenous injection of CM was able to significantly
increase the deposition of Col V fibers and IL-17 on both
day 14 and day 21 as compared with the amount
observed in rats from the BLM group and MSC groups.
In conclusion, this study reinforces previous observations
on the therapeutic properties of MSCs and CM and is the
first report to demonstrate the association of its actions
with immunomodulatory biomarkers on lung tissue. We
concluded that adipose-derived stem cells and adipose-
derived stem cells-CM modulate an in situ imbalance
between collagen I- and Col V-mediated IL-17 immune
response, emerging as a promising therapeutic option for
recovering from BLM pulmonary fibrosis2020-01-01T00:00:00ZDistribution and microstructure of intrarenal arteries in Bactrian camels (Camelus bactrianus)
http://hdl.handle.net/10201/125389
Título: Distribution and microstructure of intrarenal arteries in Bactrian camels (Camelus bactrianus)
Autor/es principal/es: Li, Hui; Cui, Yan; Wang, Yali; Qiu, Haiyu; Afedo, Seth Yaw; Huang, Yufeng; Bai, Xuefeng
Resumen: Studies and reports focusing on the Bactrian
camels’ kidney structure from an anatomical perspective
are scanty, therefore, this work aims to systemically
investigate the anatomical structure of the kidney and
examine the distribution and microstructure of intrarenal
arteries. Ten pairs of healthy adult kidneys from male
and female Bactrian camels were used in the study. The
kidney of Bactrian camel appeared like a broad bean
with a smooth surface. Using artery casting, we observed
that the renal artery divided into dorsal and ventral
branches; the dorsal branch continuously divided into a
shorter anterior branch and a longer posterior branch,
while the ventral branch directly divided into interlobar
arteries. The number of interlobar arteries in the left and
right kidneys were slightly different, 14 to 16 in left
while 16 in the right kidney. No anastomosis was found
between the dorsal and ventral branches or their sub-
branches. To further study the microscopic structure,
microanatomy and scanning microscope were used.
Surprisingly, we observed two other ways afferent
arteriole arose apart from the interlobular artery. They
were the arcuate artery and conjoint afferent arteriole.
Two afferent arterioles supplied one glomerulus and
occasionally the absence of glomerulus was also
observed, where the arteriole kept extending, and no
typical glomerulus formed. Since branching of arteries
and urologic function of kidneys are physiologically
integrated, these features of Bactrian camel may help to
further investigate their adaptations to desert climate.2020-01-01T00:00:00Z