156 research outputs found
nâ3 PUFA dietary supplementation inhibits proliferation and store-operated calcium influx in thymoma cells growing in Balb/c mice
The antitumor effect of daily individual administration of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (2 g/kg body weight) in Balb/c mice bearing a transplantable thymoma was investigated. Mice received oleic acid (control group), EPA and DHA ethyl esters starting 10 days before tumor inoculation. Analysis of phospholipid composition of neoplastic cell revealed that EPA and DHA levels were significantly increased (63 and 22% increase) after EPA and DHA treatments, respectively. Conversely, decreased levels of arachidonic acid were found in both cases (19 and 24% decrease in EPA and DHA groups, respectively). EPA and DHA delayed the appearance of macroscopic ascites (100% of animal, from 7 to 28 days), prolonged animal survival (100% of animal, from 22 to 32 and 33 days, respectively) and reduced the percentage of proliferating tumor cells detected by immunostaining of proliferation cell nuclear antigen (PCNA) (80 and 85% decrease, respectively). Moreover, the regulatory effects of these dietary nâ3 fatty acids on the influx of Ca2+, activated by depletion of intracellular stores with thapsigargin (Tg), were investigated. By using a Ca2+-free/Ca2+-reintroduction protocol and Fura-2 as fluorescent indicator of intracellular free Ca2+([Ca2+]i), we observed that EPA and DHA treatments markedly decreased Tg-induced rise in [Ca2+]i (49 and 37% decrease, respectively). This effect was related to the inhibition of the store-operated Ca2+ influx, as confirmed also by Mn2+ influx experiments. The inhibitory action of EPA and DHA on the store-operated Ca2+ influx could explain, at least in part, their antitumoral activity, as this Ca2+ mobilization pathway appears to be involved in the cell signaling occurring in non-excitable cells to evoke many cellular processes, including cell proliferation. âCalviello, G., P. Palozza, F. Di Nicuolo, N. Maggiano, and G. M. Bartoli. Nâ3 PUFA dietary supplementation inhibits proliferation and store-operated calcium influx in thymoma cells growing in Balb/c mice
Evaluation of the Siemens HIV Antigen-Antibody Immunoassay
Fourth-generation assays for the simultaneous detection of human immunodeficiency virus (HIV) antigen and antibodies are available on the international market and are currently used for blood donor screening and for HIV diagnosis. In this study we evaluated the performance of the novel automated fourth-generation ADVIA CentaurÂź HIV Ag/Ab Combo assay. The assay detected seroconversion at the same bleed or at least one bleed earlier in panels with respect to other assays and showed a detection efficacy equal to those of other assays in a low-titer panel. Samples obtained from blood donors (n = 2,778) or from HIV-positive patients (HIV-1 B subtype, n = 82; non-B subtype, n = 71) were also tested, showing a good correlation with other fourth-generation assays. We assessed the performance of 3 fourth-generation assays for detecting in utero transmitted anti-HIV antibodies and found a more specific detection efficiency with the ADVIA Centaur HIV Ag/Ab Combo assay compared to the other fourth-generation assays
Synthetic PreImplantation Factor (sPIF) reduces inflammation and prevents preterm birth.
Preterm birth (PTB) is the leading cause of neonatal morbidity and mortality and spontaneous PTB is a major contributor. The preceding inflammation/infection contributes not only to spontaneous PTB but is associated with neonatal morbidities including impaired brain development. Therefore, control of exaggerated immune response during pregnancy is an attractive strategy. A potential candidate is synthetic PreImplantation Factor (sPIF) as sPIF prevents inflammatory induced fetal loss and has neuroprotective properties. Here, we tested maternal sPIF prophylaxis in pregnant mice subjected to a lipopolysaccharides (LPS) insult, which results in PTB. Additionally, we evaluated sPIF effects in placental and microglial cell lines. Maternal sPIF application reduced the LPS induced PTB rate significantly. Consequently, sPIF reduced microglial activation (Iba-1 positive cells) and preserved neuronal migration (Cux-2 positive cells) in fetal brains. In fetal brain lysates sPIF decreased IL-6 and INFγ concentrations. In-vitro, sPIF reduced Iba1 and TNFα expression in microglial cells and reduced the expression of pro-apoptotic (Bad and Bax) and inflammatory (IL-6 and NLRP4) genes in placental cell lines. Together, maternal sPIF prophylaxis prevents PTB in part by controlling exaggerated immune response. Given the sPIF`FDA Fast Track approval in non-pregnant subjects, we envision sPIF therapy in pregnancy
Resistin modulates glucose uptake and glucose transporter-1 (GLUT-1) expression in trophoblast cells
The adipocytokine resistin impairs glucose tolerance and insulin sensitivity. Here, we examine the effect of resistin on glucose uptake in
human trophoblast cells and we demonstrate that transplacental glucose transport is mediated by glucose transporter (GLUT)-1.
Furthermore, we evaluate the type of signal transduction induced by resistin in GLUT-1 regulation. BeWo choriocarcinoma cells and primary
cytotrophoblast cells were cultured with increasing resistin concentrations for 24 hrs. The main outcome measures include glucose
transport assay using [3H]-2-deoxy glucose, GLUT-1 protein expression by Western blot analysis and GLUT-1 mRNA detection by quantitative
real-time RT-PCR. Quantitative determination of phospho(p)-ERK1/2 in cell lysates was performed by an Enzyme Immunometric
Assay and Western blot analysis. Our data demonstrate a direct effect of resistin on normal cytotrophoblastic and on BeWo cells: resistin
modulates glucose uptake, GLUT-1 messenger ribonucleic acid (mRNA) and protein expression in placental cells. We suggest that ERK1/2
phosphorylation is involved in the GLUT-1 regulation induced by resistin. In conclusion, resistin causes activation of both the ERK1 and
2 pathway in trophoblast cells. ERK1 and 2 activation stimulated GLUT-1 synthesis and resulted in increase of placental glucose uptake.
High resistin levels (50â100 ng/ml) seem able to affect glucose-uptake, presumably by decreasing the cell surface glucose transporter
Identification of Novel Putative Urinary Markers of Endometriosis by High-Resolution Quantitative Proteomics
Endometriosis is a chronic gynecological inflammatory disease characterized by the presence of functional endometrial glands..
Effect of alphaâlipoic acid and myoinositol on endometrial inflammasome from recurrent pregnancy loss women
Problem: A significant increased expression/activation of one of the most wellâcharacterized inflammasomes, the NAcht leucineârichârepeat proteinâ3 (NALPâ3), in the endometrium from idiopathic recurrent pregnancy loss women (RPL) has been previously found by our research group. We therefore, suggested this event as being one of the molecular mechanisms altering endometrial inflammatory status during early pregnancy. In the present research, we attempt to investigate whether molecules with antiâinflammatory activity, alphaâlipoic acid (ALA), and/or myoinositol affect the
endometrial NALPâ3 expression and activation. Method of study: Women with a history of idiopathic RPL (n = 30) were included in the study and compared to a control group (n = 15). Endometrial tissues were collected by hysteroscopy during the midâluteal phase. RPL women underwent a threeâmonth prescription of tablets containing ALA plus myoinositol (SinopolÂź). After
treatment, hysteroscopic biopsies were repeated in RPL patients. Inflammasome expression was evaluated by immunohistochemical and Western blot analysis. NALPâ3 activation was studied by quantifying the secretion of both caspaseâ1 and interleukin IL)â1Ă and ILâ18 through ELISA. In ex vivo experiments, the effects of each molecule on endometrial inflammasome were studied.
Results: SinopolÂź significantly reduced the RPL endometrial inflammasome expression and activation. ALA, but not myoinositol, significantly reduced the endometrial inflammasome expression and activity. Conclusion: Our data suggest a role for ALA on RPL inflammasome. Understanding the mechanisms involved in RPL and the observation that specific molecules are able
to interfere with such complex at the endometrium might provide new rational design approaches to a personalized evaluation of endometrial status and, ultimately, a targeted medicine
Synthetic PreImplantation Factor (PIF) prevents fetal loss by modulating LPS induced inflammatory response
Maternal control of inflammation is essential during pregnancy and an exaggerated
response is one of the underlying causes of fetal loss. Inflammatory response is mediated
by multiple factors and Toll-like receptors (TLRs) are central. Activation of TLRs results in
NALP-3 mediated assembly of apoptosis-associated speck-like protein containing a CARD
(ASC) and caspase-1 into the inflammasome and production of pro-inflammatory cytokines
IL-1\u3b2 and IL-18. Given that preventing measures are lacking, we investigated PreImplantation
Factor (PIF) as therapeutic option as PIF modulates Inflammation in pregnancy. Additionally,
synthetic PIF (PIF analog) protects against multiple immune disorders. We used
a LPS induced murine model of fetal loss and synthetic PIF reduced this fetal loss and
increased the embryo weight significantly. We detected increased PIF expression in the placentae
after LPS insult. The LPS induced serum and placenta cytokines were abolished by
synthetic PIF treatment and importantly synthetic PIF modulated key members of inflammasome
complex NALP-3, ASC, and caspase-1 as well. In conclusion our results indicate that
synthetic PIF protects against LPS induced fetal loss, likely through modulation of inflammatory
response especially the inflammasome complex. Given that synthetic PIF is currently
tested in autoimmune diseases of non-pregnant subjects (clinicaltrials.gov, NCT02239562),
therapeutic approach during pregnancy can be envisioned
Low-molecular-weight heparins induce decidual heparin-binding epidermal growth factorâlike growth factor expression and promote survival of decidual cells undergoing apoptosis Nicoletta Di
Objective: To evaluate the effects of low-molecular-weight heparins (LMWHs) on decidual heparin-binding epidermal growth factorâlike growth factor
(HB-EGF) expression/secretion and on TNF-aâinduced decidual apoptosis.
Design: Experimental study.
Setting: Department of Obstetrics and Gynecology, Universita Cattolica del Sacro Cuore, Rome, Italy.
Patient(s): Cultures of primary decidual cells isolated from human term placenta.
Intervention(s): The effects of LMWHs (tinzaparin and enoxaparin) on decidual HB-EGF expression and secretion were investigated by Western blot
analysis and ELISA, respectively. TNF-aâinduced decidual apoptosis was evaluated by annexin V staining, terminal deoxynucleotide transferaseâ
mediated dUTP nick-end labeling (TUNEL) assay, and caspase activities.
Main Outcome Measure(s): Decidual HB-EGF expression/secretion and apoptotic rate induced by TNF-a were investigated.
Result(s): Tinzaparin enhanced decidual HB-EGF expression and secretion. TNF-a reduced the number of viable cells by inducing apoptosis.
Simultaneous addition of LMWHs (primarily tinzaparin) blocked the increase in annexin Vâ and TUNEL-positive cells and reduced the amount of
caspase activities.
Conclusion(s): Both LMWHs induced a significant increase in decidual HB-EGF expression/secretion and reduced TNF-aâinduced decidual apoptosis.
Tinzaparin demonstrated higher efficacy. (Fertil Steril 2012;97:169â77. 2012 by American Society for Reproductiv
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