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Abstract

Coronary artery bypass grafting (CABG), together with percutaneous coronary intervention (PCI), are both still the most efficient procedures for myocardial revascularization to treat advanced coronary artery disease (CAD). Donor blood vessels used in CABG are usually the internal thoracic artery (ITA) and saphenous vein (SV). The importance of inflammation and inflammatory pathways in graft patency is well established. Nevertheless, not all molecular mechanisms underlying the inflammatory process appear to be clear. Employing the expressive microarray approach to analyze the transcriptome of both venous and arterial grafts, five GO BP terms has been selected: “cellular response to interferon-gamma”, “inflammatory response”, “interferon-gamma-mediated signaling pathway”, “response to interferon-gamma” and “positive regulation of inflammatory response”. This study aimed to evaluate potential molecular factors that could be characteristic markers for both SV and ITA conduits.

Running title: Aortocoronary conduits may show a different inflammatory response

Abstract

In vitro models represent an alternative technique to in vivo or ex vivo studies in the drug development process. Cell-based assays are used to measure the level of proliferation and toxicity, as well as activation of signalling pathways and changes in morphology in cultivated cells. The studies conducted in vitro are aimed to estimate the newly synthesised drugs’ ability to permeate biological barriers and exert their therapeutic or cytotoxic effects. However, more than half of all studied drugs fail in the second or third phase of clinical trials due to a lack of confirmed efficacy. About a third of drugs fail because of safety issues, such as unacceptable levels of toxicity. To reduce attrition level in drug development, it is crucial to consider the implementation of translational phenotypic assays as well as to decipher various molecular mechanisms of action for new molecular entities. In this review, we summarise the existing cell-based methods most frequently used in the studies on drugs, taking into account their advantages and drawbacks.

Running title: Cell-based approaches in drug development

Abstract

Coronary artery bypass graft (CABG) is the surgical method most commonly used to treat coronary artery disease (CAD). The vessels that are used in CABG are usually the internal thoracic artery (ITA) and the saphenous vein (SV). Transplant patency is one of the most important factors affecting transplant success. In this study, we used an expressive microarray method, approved by RT-qPCR, for transcriptome analysis of arterial and venous grafts. In the search for potential molecular factors, we analyzed gene ontologies of different expression based on the muscular system. Among interesting groups, we distinguished muscle cell proliferation, muscle contraction, muscle system process, regulation of smooth muscle cell proliferation and smooth muscle cell proliferation. The highest increase in gene expression was observed in: ACTN2, RBPMS2, NR4A3, KCNA5, while the smallest decrease in expression was shown by the P2RX1, KCNH2, DES and MYOT genes. Particularly noteworthy are the ACTN2 and NR4A3 genes, which can have a significant impact on vascular patency. ACTN2 is a gene that can affect the formation of atherosclerotic plaques, while NR4A3 occurs in 4 of the 5 ontological groups discussed and can affect the inflammatory process in the blood vessel. To summarize, the presented study provided valuable insight into the molecular aspects characterizing the vessels used in CABG, and in particular identified genes that may be the target for further studies on duct patency.

Running title: CABG grafts’ molecular analysis of ‘muscle system process’

Abstract

The presence of CCs around the oocyte after ovulation is one of the key elements contributing to oocyte developmental competence. In the presented study, we used CCs from 12 patients aged 18-40 diagnosed with infertility. After harvesting cells on day 1, 7, 15 and 30 of culture, total RNA was isolated and transcriptomic analysis was performed. The DAVID software indicated the following GO BP terms: “cell junction organization”, “cell migration”, “cell morphogenesis involved in differentiation”, “cell morphogenesis” and “cell motility”. Of the genes belonging to all ontological groups, the most downregulated were: SLC7A8, DFNB31, COL1A1, CDC42SE1, TGFBR3, HMGB1, with the most upregulated genes being: ANXA3, KIAA1199, HTR2B, VCAM1, DKK1.

While many studies focus on attempts to obtain fully competent oocytes, scientists still have difficulty attaining adequate results in vitro. Lack of adequate knowledge often results in low in vitro fertilization efficiency. Therefore, our research focuses on CCs cells, thanks to which the oocyte most likely acquires developmental competence. The main purpose of the study was to identify the potential molecular markers responsible for cell junction organization, migration, differentiation, morphogenesis and motility.

Running title: New markers of human cumulus oophorus cells cultured in vitro

Abstract

The growth and maturation of the oocyte is a dynamic process which requires a variable supply of hormones, growth factors and energy. These needs are met partially by the surrounding somatic cells and the cumulus-oocyte complex, which communicate bi-directionally via gap junctions. Identifying and analyzing protein expression in the oocyte can provide insight in its development and growth. Further, like bone marrow stem cells, if relevant marker genes are found in oocytes, there is a potential for the oocyte to be manipulated into becoming hemopoietic stem cells. In this study, porcine oocytes were isolated and subjected to microarray analysis to compare the oocyte gene expression in vivo and in vitro maturation (IVM). Genes identified belonged to both ‘hemopoietic or lymphoid organ development’(GO:0048534) and ‘immune system development’ (GO:0002520), and the markers can be used to identify several activities such as cell migration, neurogenesis and proliferation. The following are the identified genes and all were downregulated after IVM to varying degrees: ID2, VEGFA, TGFBR3, INHBA, CDK6, BCL11A, MYO1E, ITGB1, EGR1, NOTCH2, SPTA1, KIT and TPD52. Our results should provide new markers to further investigate oocyte development and growth regulation.

Running title: Markers of hemopoietic organ development

Abstract

Mucous membranes appear in various parts of the whole body performing similar functions. However, they differ based on where the mucosa is located. It functions as a barrier in such systems as: respiratory, urogenital and digestive . In this study we will be focusing strictly on the oral mucosa. Keratinocytes and fibroblasts, which mainly form the structure of the oral mucosa, are subjected to numerous factors. Being one of the million parts that build the animal organism, they are involved in various processes. In this study, we will try to confirm that in the in vitro culture of oral mucosa cells, the expression of our selected genes undergoes significant changes which are tied to such processes as: homeostasis, chemotaxis and organic/inorganic response of the organism. For this study, 20 pubertal crossbred Landrace gilts were used. After slaughter, samples of buccal pouch mucosa were obtained and transported to the laboratory. The excised tissue was prepared and processed due to protocols. The final pellet was resuspended in supplemented DMEM. Once the cultures attained 70–80% confluency, they were passaged. Total RNA from each pooled sample was subjected to two rounds of sense cDNA amplification. The cDNA was processed on microarrays. Analysis of the scanned arrays was performed. The files were imported into downstream data analysis software. The DAVID analysis showed that differently expressed genes belongs to 56 Gene ontology groups. In this paper we focused on “cellular divalent inorganic cation homeostasis”, “chemical homeostasis”, “chemotaxis”, “homeostatic process” and “response to organic substance” GO BP terms. These sets of genes were subjected to hierarchical clusterization procedure. In summary, the data we collected showed primarily changes in gene expression that occurred in the thirty-day cell culture of oral mucosa tissue. We assume that indicated genes could be new gene markers for studied processes.

Running title: Homeostasis in oral mucosa cells

Abstract

The growth and development of the oocyte is essential for the ovarian follicle. Cumulus cells (CCs) - a population of granulosa cells - exchange metabolites, proteins and oocyte-derived paracrine factors with the oocyte through gap junctions, to contribute to the competency and health of the oocyte. This bi-directional communication of the cumulus-oocyte complex could be better understood through the micro-analysis of a porcine oocyte gene expression before in vitro maturation (IVM) and after. Additionally, the study of the somatic and gamete cells differentiation capability into neuronal lineage would be promising for future stem cell research as granulosa cells are easily accessible waste material from in vitro fertilization (IVF) procedures. Therefore, in this study, the oocytes of 45 pubertal Landrace gilts were isolated and the protein expression of the COCs were analyzed through micro-analysis techniques. Genes belonging to two ontological groups: neuron differentiation and negative regulation of cell differentiation have been identified which have roles in proliferation, migration and differentiation. Twenty identified porcine oocyte genes (VEGFA, BTG2, MCOLN3, EGR2, TGFBR3, GJA1, FST, CTNNA2, RTN4, MDGA1, KIT, RYK, NOTCH2, RORA, SMAD4, ITGB1, SEMA5A, SMARCA1, WWTR1 and APP) were found to be down-regulated after the transition of IVM compared to in vitro. These results could be applied as gene markers for the proliferation, migration and differentiation occurring in the bi-directional communication between the oocyte and CCs.

Running title: Differentiation and neurogenesis in oocyte cells

Abstract

The increasing resistance against classical antibiotic treatment forces the researchers to develop novel non-toxic antimicrobial agents. The aim of this study was to determine the antimicrobial properties of seven different porphyrins having distinctive hydrophobicity/hydrophilicity: P1 meso-tetra(4-methoxy-phenyl)porphyrin, P2 Zn(II)-meso-5,10,15,20-tetrapyridylporphyrin, P3 meso-tetra(p-tolyl)porphyrin, P4 5,10,15,20-tetraphenylporphyrin; P5 (5,10,15,20-tetraphenylporphinato) dichlorophosphorus(V) chloride, P6 5,10,15,20-tetrakis-(N-methyl-4-pyridyl) porphyrin-Zn(II) tetrachloride, P7 Zn(II)-5,10,15,20-meso-tetrakis-(4-aminophenyl)porphyrin. The meso-porphyrin derivatives were screened for their antimicrobial activity against six reference strains: Streptococcus pyogenes ATCC 19615, Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Klebsiella pneumoniae ATCC 700603, Pseudomonas aeruginosa ATCC27853 and Candida albicans ATCC 10231. The antimicrobial activity of these samples was evaluated by the agar disk diffusion method and dilution method, with the determination of the minimum inhibitory concentration (MIC), the minimum bactericidal concentration (MBC) and the minimum fungicidal concentration (MFC). The most significant result is provided by the water-soluble P5 manifesting an obvious antimicrobial activity against Streptococcus pyogenes. On the other hand, P6 is a moderately active derivative against Streptococcus pyogenes and Escherichia coli and P7 presents moderate activity against Streptococcus pyogenes and Staphylococcus aureus. All the tested porphyrin bases, presenting hydrophobic character, have no antimicrobial activity under the investigated conditions. The common characteristics of the porphyrins that act as promising antimicrobial agents in the non-irradiated methods are: the cationic nature, the increased hydrophilicity and the presence of both amino functional groups grafted on the porphyrin ring and the coordination with Zn or phosphorus in the inner core.

Abstract

Aim: We aimed to examine the association between several circulating bone turnover markers [ osteocalcin (OC), osteoprotegerin (OPG), beta-CrossLaps (β-CTx)], hip and spine bone mineral density (BMD) and abdominal aortic calcification (AAC) in patients with chondrocalcinosis (CC).

Methods: Thirty-six patients with CC and thirty-seven controls were consecutively enrolled in this pilot case-control, cross-sectional study. The following parameters were assessed: serum levels of OC, OPG and β-CTx by enzyme-linked immunosorbent assay (ELISA); hip and spine BMD by dual-energy X-ray absorptiometry and AAC score by lateral radiography.

Results: Patients with CC had higher levels of serum bone turnover markers and AAC score than the control group: OC [6.5 (3.5-9.9) vs 4.5 (2.6-7.2) ng/ml; p=0.05], OPG [(7.7 (6.2-9.4) vs 6.5 (5.5-8.12) pmol/ml; p=0.02], β-CTx [6078 (5870-6171) vs 5851 (5465-6109) pg/ml; p=0.02] and AAC score (3.6±6.2 vs 0.5±2; p=0.006). Conversely, even if statistical significance was not reached, hip and spine BMD was lower in patients with CC. Additionally, we found a positive correlation between OPG and AAC, but also between OPG and osteoporosis in patients with CC.

Conclusion: Patients with CC are characterized by higher circulating OC, OPG and β-CTx. The presence of AAC was more common in patients with CC, being only associated with serum OPG.

Abstract

Acute lymphoblastic leukemia is the most common hematological malignancy at pediatric age. Cardiotoxicity holds the first place among the causes of morbidity and mortality in these patients. Anthracyclines are cytostatic drugs frequently associated with cardiotoxicity. Early diagnosis of cardiac impairment during the treatment of pediatric patients is extremely important, both for modulating future chemotherapy and for administering cardioprotective agents. Long term monitoring after chemotherapy helps to identify the risk of late cardiotoxicity among cancer survivors. There are several biomarkers, already in use or still under study, which may represent an operator-independent alternative for echocardiography in the diagnosis of cardiotoxicity. In case of cardiac damage, the clinician has options for treating or limiting the progression, either with the use of already approved agents, such as Dexrazoxane, or by administrating other cardioprotective drugs. International experts are still attempting to establish the best algorithm for early detection of cardiotoxicity, as well as the most efficient treatment plan in case of already existing myocardial damage in these patients. We present a review on treatment-related cardiotoxicity, including mechanisms of development, useful biomarkers and treatment options, after carefully analyzing specialty literature.