Liliana Paslaru

Heat shock factor 2 is a stress‐responsive mediator of neuronal migration defects in models of fetal alcohol syndrome

Fetal alcohol spectrum disorder (FASD) is a frequent cause of mental retardation. However, the molecular mechanisms underlying brain development defects induced by maternal alcohol consumption during pregnancy are unclear. We used normal and Hsf2‐deficient mice and cell systems to uncover a pivotal role for heat shock factor 2 (HSF2) in radial neuronal migration defects in the cortex, a hallmark of fetal alcohol exposure. Upon fetal alcohol exposure, HSF2 is essential for the triggering of HSF1 activation, which is accompanied by distinctive post‐translational modifications, and HSF2 steers the formation of atypical alcohol‐specific HSF1–HSF2 heterocomplexes. This perturbs the in vivo binding of HSF2 to heat shock elements (HSEs) in genes that control neuronal migration in normal conditions, such as p35 or the MAPs (microtubule‐associated proteins, such as Dclk1 and Dcx), and alters their expression. In the absence of HSF2, migration defects as well as alterations in gene expression are reduced. Thus, HSF2, as a sensor for alcohol stress in the fetal brain, acts as a mediator of the neuronal migration defects associated with FASD.

Phenotypic characterization of mouse embryonic fibroblasts lacking heat shock factor 2

In murine cells, the heat shock response is regulated by a transcription factor, HSF1, which triggers the transcription of heat shock genes. HSF2 has been shown to be involved in meiosis and mouse brain development. We characterized the effects of the absence of HSF2 in mouse embryonic fibroblasts (MEFs). The temperature threshold of the heat shock response appeared lowered in Hsf2‐/‐ MEFS as monitored by the synthesis of heat shock protein HSP70. In contrast to unstressed wild type MEFS, HSP70 and HSF1 are localized in the nucleus of unstressed Hsf2‐/‐ MEFS, a characteristic of stressed cells. HSF1 is not activated for DNA‐binding at unstressed temperature in Hsf2‐/‐ MEFS. Therefore, the absence of HSF2 induces some but not all of the characteristics of the stress response. In addition, Hsf2‐/‐ MEFS exhibited proliferation defects, altered morphology, remodeling of the fibronectin network.

Assessment of Gene Expression Profiles in Peripheral Occlusive Arterial Disease

BACKGROUND Molecular events responsible for the onset and progression of peripheral occlusive arterial disease (POAD) are incompletely understood. Gene expression profiling may point out relevant features of the disease. METHODS Tissue samples were collected as operatory waste from a total of 36 patients with (n = 18) and without (n = 18) POAD. The tissues were histologically evaluated, and the patients with POAD were classified according to Leriche-Fontaine (LF) classification: 11% with stage IIB, 22% with stage III, and 67% with stage IV. Total RNA was isolated from all samples and hybridized onto Agilent 4×44K Oligo microarray slides. The bioinformatic analysis identified genes differentially expressed between control and pathologic tissues. Ten genes with a fold change ≥ 2 (1 with a fold change ≥ 1.8) were selected for quantitative polymerase chain reaction validation (GPC3, CFD, GDF10, ITLN1, TSPAN8, MMP28, NNMT, SERPINA5, LUM, and FDXR). C-reactive protein (CRP) was assessed with a specific assay, while nicotinamide N-methyltransferase (NNMT) was evaluated in the patient serum by enzyme-linked immunosorbent assay. RESULTS A multiple regression analysis showed that the level of CRP in the serum is correlated with the POAD LF stages (r(2) = 0.22, P = 0.046) and that serum NNMT is higher in IV LF POAD patients (P = 0.005). The mRNA gene expression of LUM is correlated with the LF stage (r(2) = 0.45, P = 0.009), and the mRNA level of ITLN1 is correlated with the ankle-brachial index (r(2) = 0.42, P = 0.008). CONCLUSIONS Our analysis shows that NNMT, ITLN1, LUM, CFD, and TSPAN8 in combination with other known markers, such as CRP, could be evaluated as a panel of biomarkers of POAD.

The effect of Cyclosporine A on cardiomyocytes differentiation

Cyclosporine A (CsA) is a powerful immunosuppressive drug which significantly improved the success of organ transplantation; however, the major limiting factors for the drugs clinical use are its long and short term adverse effects. The present study was conducted to examine, in a dose‐dependent manner, in a model of cardiogenesis, the effect of CsA on cardiomyocytes differentiation.

Optical manipulation of Saccharomyces cerevisiae cells reveals that green light protection against UV irradiation is favored by low Ca2+ and requires intact UPR pathway

Optical manipulation of Saccharomyces cerevisiae cells with high density green photons conferred protection against the deleterious effects of UV radiation. Combining chemical screening with UV irradiation of yeast cells, it was noted that the high density green photons relied on the presence of intact unfolded protein response (UPR) pathway to exert their protective effect and that the low Ca2+ conditions boosted the effect. UPR chemical inducers tunicamycin, dithiotreitol and calcium chelators augmented the green light effect in a synergic action against UV‐induced damage. Photo‐manipulation of cells was a critical factor since the maximum protection was achieved only when cells were pre‐exposed to green light.

Evaluation of Tumor Response Using Alpha-fetoprotein and Des-gamma-carboxy Prothrombin in Hepatocellular Carcinoma Patients Who Underwent Transarterial Chemoembolization

Introduction: The aim of the study is to evaluate the role of alpha-fetoprotein (AFP) and des-y-carboxy prothrombin (DCP) in the assessment of treatment response at one month in patients with hepatocellular carcinoma (HCC) treated with trans-arterial chemoembolization (TACE). Methods: From March 2016 to April 2017 a number of 59 patients diagnosed with HCC were prospectively enrolled. A TACE procedure as initial treatment modality was performed in 41 patients. AFP and DCP serum levels were measured and clinical features were determined for all the patients that were included. The Wilcoxon rank test was used to compare variables at baseline and at one month after the procedure. Results: Treatment was performed in 86.4% of the patients diagnosed with HCC, 27 patients received a classical TACE procedure, 14 patients were treated with DEB-TACE, radiofrequency ablation was performed in 3 patients and 4 patients received a liver transplant as initial treatment. Systemic therapy with Sorafenib was started in 3 patients (5%) and in 8 cases no treatment was performed. AFP value significantly decreased at one month in patients that underwent TACE therapy (median value 240.3 vs. 123.7 ng/mL, p=0.020). The same significant decrease was noted for DCP values (median value 1376.8 vs. 769 mAU/mL, p=0.0033). Both AFP (85.5 vs. 18.7 ng/mL, p=0.035) and DCP values (693.2 vs. 58.2 ng/mL, p=0.0003) were significantly lower only in subjects who achieved complete response after TACE and not in patients with partial response. In patients treated with TACE therapy, there was a down-sizing of the maximum diameter of the tumor nodule (30 vs. 27 mm, p=0.02). CONCLUSION There was a significant decrease of AFP and DCP values after complete response in HCC patients treated with TACE. DCP is a more effective tumor marker in predicting response than AFP, with no benefit found in their combination.

The Romanian National Program for Liver Transplantation - 852 Procedures in 815 Patients over 17 Years (2000-2017): A Continuous Evolution to Success

Background: Liver transplantation (LT) has become an established treatment for end-stage liver disease, with more than 20.000 procedures yearly worldwide. The aim of this study was to analyze the results of Romanian National Program of LT. Methods: Between April 2000 and April 2017, 817 pts received 852 LTs in Romania. Male/female ratio was 487/330, while adult/pediatric ratio was 753/64, with a mean age of 46 years (median 50 yrs; range 7 months - 68 yrs). Main LT indications were HBV cirrhosis (230 pts; 28.2%), HCC (173 pts; 21.2%), and HCV cirrhosis (137 pts; 16.8%). Waiting time and indications for LT, patient and donor demographics, graft features, surgical procedures, and short and long-term outcomes were analyzed. Results: DDLT was performed in 682 pts (83.9%): whole LT in 662 pts (81%), split LT in 16 pts (2.3%), reduced LT in 2 pts (0.2%), and domino LT in 1 pts (0.1%). LDLT was performed in 135 pts (16.5%): right hemiliver in 93 pts (11.4%), left lateral section in 28 pts (3.4%), left hemiliver in 8 pts (1%), left hemiliver with segment 1 in 4 pts (0.5%), and dual graft LDLT in 2 pts (0.2%). Overall major morbidity rate was 31.4% (268 pts), while perioperative mortality was 7.9% (65 pts). Retransplantation rate was 4.3% (35 pts): 27 whole LTs, 3 reduced LTs, 3 split LTs, and 2 LDLT. Long-term overall 1-, 3-, and 5-year estimated survival rates for patients were 87.9%, 81.5%, and 79.1%, respectively. One-, 3-, and 5-year overall mortality on waiting list also decreased significantly over time from 31.4%, 54.1% and 63.5%, to 4.4%, 13.9% and 23.6%, respectively. Conclusions: The Romanian National program for liver transplantation addresses all causes of acute and chronic liver failure or liver tumors in adults and children, using all surgical techniques, with good long-term outcome. The program constantly evolved over time, leading to decreased mortality rate on the waiting list.

Effects Induced in Complex Biological Systems by High Density Green Photons

It is known that light interaction with matter may generate an optical force, which may produce modifications at the physical and chemical level. The basic technique of the field uses strongly focused laser beams that trap small objects and manipulate local structures. In our work we use irradiation of complex biological molecules with high density green photons, which may induce electric dipoles by polarization effects. The resulting dipolar interaction may lead to organized structures like molecular aggregates and microparticles. We present experimental evidence of such an optical manipulation on long alkanes chains and two specific enzymes. A preliminary physical model is suggested for acounting of these specific interaction forces.