Oviductal microvesicles and their effect on in vitro maturation of canine oocytes.

The effect of conditioned medium (CM) or microvesicles (MVs), secreted by multicellular spheroids of oviductal cells, and the involvement of some microRNAs (miRNAs) were investigated in canine oocyte maturation. To generate CM, spheroids were cultured for 3 days. MVs were obtained by ultracentrifugation of CM at 100,000 g and measured for size and concentration by NanoSight instrument. Cumulus-oocyte complexes (COCs) were matured at 38.5°C with 5% CO2 and 5% of O2 in synthetic oviductal fluid (SOF) in biphasic systems: for 24 h, with 5.0 µg/mL of LH and for other 48 h with 10% oestrous bitch serum. SOF was used as control (CTR) or supplemented with 10% CM or 25-50-75-100-150 × 106 MVs/mL labeled with PKH-26. Results show that multicellular aggregates secreted shedding vesicles. By fluorescence microscopy, the incorporation of labeled MVs was visible only at 72 h in oocyte cytoplasm. These MVs had a positive effect (P < 0.05) on maturation rate (MII) at the concentration of 75 and 100 × 106 MVs/mL compared to CM and CTR (20.34% and 21.82% vs 9.09% and 8.66% respectively). The concentration of 150 × 106 MVs/mL provided only 9.26% of MII. The expression of three specific miRNAs (miR-30b, miR-375 and miR-503) was studied. The lower rate of MII with the higher concentration of MVs is possibly due to the high level of miR-375. In conclusion, the oviductal MVs could be involved in cellular trafficking during oocyte maturation and their possible use in vitro could facilitate the exploitment of canine reproductive biotechnologies.

Assessing liver function: diagnostic efficacy of parenchymal enhancement and liver volume ratio of Gd-EOB-DTPA-enhanced MRI study during interstitial and hepatobiliary phase.

AIM: To assess the efficacy of signal intensity in interstitial and hepatobiliary phase normalized for liver volume, on gadolinium-ethoxybenzyl-diethylenetriaminepentaacetic acid (Gd-EOB-DTPA)-enhanced magnetic resonance imaging (MRI) study, for the evaluation of liver function through the comparison with Child-Pugh (CP), model for end-stage liver disease (MELD), and biochemical tests. METHODS: All dynamic Gd-EOB-DTPA MRI studies performed in patients with suspected liver lesions were retrospectively reviewed. The rate of liver-to-muscle ratio on T1 sequence 70 s (interstitial phase) and 20 min (hepatobiliary phase) after injection of Gd-EOB-DTPA was calculated for each MRI study and then normalized for liver volume (irINTnorm and irHEPnorm). Pearson correlation coefficient was computed to assess the correlation among these values and CP and MELD scores, and biochemical tests. RESULTS: A total of 303 MRI studies, performed on 221 patients, were included. Mean age was 63.8 years ± 12.9 with a majority of male patients (186; 61.4%). A total of 186 out of 303 (61.4%) were cirrhotic patients. The irHEPnorm was significantly lower in cirrhotic than non-cirrhotic patients (0.0004 ± 0.0002 to 0.0005 ± 0.0003, p = 0.010). This value had a moderate, significant correlation with Child-Pugh and MELD scores (R = - 0.292, p < 0.0001 and R = - 0.192, p = 0.010, respectively). In particular, irHEPnorm progressively decreased from Child-Pugh A to C (0.0004-0.0002, p < 0.0001) and from MELD ≤ 10 to 19-24 (0.0004-0.0003, p = 0.018). Among biochemical parameters, total bilirubin, GOT, and albumin had the strongest correlation with irHEPnorm (R = - 0.258, - 0.291, and 0.262, p < 0.0001, respectively). No correlations were found between irINTnorm and CP and MELD scores. CONCLUSION: irHEPnorm value derived from Gd-EOB-DTPA-enhanced MRI is a reliable, non-invasive, useful tool to quantify liver function and to assess the degree of cirrhosis, offering a strict relationship with clinical scores and biochemical parameters. This could help surgeons in clinical decision-making, allowing them to choose the more suitable surgical approach for cirrhotic patients.

Estimating liver function in a large cirrhotic cohort: Signal intensity of gadolinium-ethoxybenzyl-diethylenetriamine penta-acetic acid-enhanced MRI.

BACKGROUND: To assess whether gadolinium-ethoxybenzyl-diethylenetriamine penta-acetic acid-enhanced MRI study is useful to estimate liver function in comparison to the presence or absence of cirrhosis, Child Pugh (CP), Model for End-stage Liver Disease (MELD), ALBI scores and biochemical test. METHODS: We retrospectively reviewed all consecutive Gd-EOB-DTPA-enhanced-MRI studies performed between 2010 and 2016 in patients with focal liver lesions undergoing clinical evaluation. Patients were divided in study and control group according to the presence of cirrhosis, and then classified by CP, MELD and ALBI. Signal intensity was calculated through the liver-to-muscle ratio in portal- (SI-POR) and hepatobiliary-phase(SI-HEP). RESULTS: Three-hundred-three Gd-EOB-DTPA liver-enhanced-MRI studies were included. One-hundred-ninety-one patients (63%) were cirrhotic. SI-HEP was significantly lower in cirrhotic group (0.55 ±â€¯0.29 vs 0.66 ±â€¯0.40, p = 0.004).The SI-HEP progressively decreased from CP-A to CP-C (0.59 ±â€¯0.28 to 0.25 ±â€¯0.19, p < 0.0001) and a significant difference was found between MELD ≤ 9 and MELD > 9 groups (0.61 ±â€¯0.31 vs 0.49 ±â€¯0.28, p = 0.007). No differences between ALBI grades were evident. Among biochemical parameters a moderate correlation was found among SI-HEP and total bilirubin, AST and albumin. CONCLUSION: SI-HEP after Gd-EOB-DTPA-enhanced-MRI effectively stratified patients with different Child Pugh grades and MELD scores. This technique could hence be useful as a novel radiological marker to estimate the underlying liver function.

Dynamic contrast enhanced perfusion CT imaging: A diagnostic biomarker tool for survival prediction of tumour response to antiangiogenetic treatment in patients with advanced HCC lesions.

PURPOSE: To investigate whether perfusion-CT (p-CT) imaging could depict the inhibition of tumor neoangiogenesis induced by Sorafenib in advanced hepatocellular carcinoma (HCC), and whether it could be useful in predicting survival during treatment. MATERIALS AND METHODS: Ninety-eight p-CT examinations were performed among 29 cirrhotic patients, with advanced HCC, before and every 2 months after Sorafenib administration, on a 256-slice MDCT scanner. Perfusion parameters were considered and statistically compared, at baseline and follow-up, between non-progressor (complete response, stable disease or partial response) and progressor (progressive disease) group. Kaplan-Meier analyses estimated the time-to-survival in overall population, after stratifying patients according to mRECIST. RESULTS: The group that responded to Sorafenib showed a significant reduction of values in HCC target lesions after anti-angiogenic therapy (p < 0.01), in comparison with progressor group that demonstrated an increase or no significant variation. When patients were stratified into mRECIST, higher survival rate was observed in the non-progressor group compared to the progressor (48.6% vs 28.6%), and statistically significant correlation (p=0.01) was found between percentage variation of perfusion parameters, from baseline to follow-up, and overall survival rate. CONCLUSION: Quantitative analysis of perfusion parameters, represents prognostic indicators useful in assessment of response to anti-angiogenic therapy, allowing for optimization of individualized treatment.