Animal Cell Culture Research Articles

Evaluating Neurodevelopmental Consequences of Perinatal Exposure to Antiretroviral Drugs: Current Challenges and New Approaches.

As antiretroviral therapy (ART) becomes increasingly affordable and accessible to women of childbearing age across the globe, the number of children who are exposed to Human Immunodeficiency Viruses (HIV) but remainuninfected is on the rise, almost all of whom were also exposed to ART perinatally. Although ART has successfully aided in the decline of mother-to-child-transmission of HIV, the long-term effects of in utero exposure to ART on fetal and postnatal neurodevelopment remain unclear. Evaluating the safety and efficacy of therapeutic drugs for pregnant women is a challenge due to the historic limitations on their inclusion in clinical trials and the dynamic physiological states during pregnancy that can alter the pharmacokinetics of drug metabolism and fetal drug exposure. Thus, much of our data on the potential consequences of ART drugs on the developing nervous system comes from preclinical animal models and clinical observational studies. In this review, we will discuss the current state of knowledge and existing approaches to investigate whether ART affects fetal brain development, and describe novel human stem cell-based strategies that may provide additional information to better predict the impact of specific drugs on the human central nervous system. Graphical Abstract Approaches to evaluate the impact of drugs on the developing brain. Dysregulation of the developing nervous system can lead to long-lasting changes. Integration of data from animal models, clinical observations, and cell culture studies is needed to predict the safety of therapeutic antiretroviral drugs during pregnancy. New approaches include human induced pluripotent stem cell (iPSC)-based 2D and 3D models of neuronal networks and brain regions, as well as single cell profiling in response to drug exposure.

Sex-specific neuroprotection by inhibition of the Y-chromosome gene, SRY, in experimental Parkinson’s disease

Parkinson's disease (PD) is a debilitating neurodegenerative disorder caused by the loss of midbrain dopamine (DA) neurons. While the cause of DA cell loss in PD is unknown, male sex is a strong risk factor. Aside from the protective actions of sex hormones in females, emerging evidence suggests that sex-chromosome genes contribute to the male bias in PD. We previously showed that the Y-chromosome gene, SRY, directly regulates adult brain function in males independent of gonadal hormone influence. SRY protein colocalizes with DA neurons in the male substantia nigra, where it regulates DA biosynthesis and voluntary movement. Here we demonstrate that nigral SRY expression is highly and persistently up-regulated in animal and human cell culture models of PD. Remarkably, lowering nigral SRY expression with antisense oligonucleotides in male rats diminished motor deficits and nigral DA cell loss in 6-hydroxydopamine (6-OHDA)-induced and rotenone-induced rat models of PD. The protective effect of the SRY antisense oligonucleotides was associated with male-specific attenuation of DNA damage, mitochondrial degradation, and neuroinflammation in the toxin-induced rat models of PD. Moreover, reducing nigral SRY expression diminished or removed the male bias in nigrostriatal degeneration, mitochondrial degradation, DNA damage, and neuroinflammation in the 6-OHDA rat model of PD, suggesting that SRY directly contributes to the sex differences in PD. These findings demonstrate that SRY directs a previously unrecognized male-specific mechanism of DA cell death and suggests that suppressing nigral Sry synthesis represents a sex-specific strategy to slow or prevent DA cell loss in PD.

An update on the detection methods of Parachlamydia acanthamoebae, an atypical agent of pneumonia

Parachlamydia acanthamoebae (P. acanthamoebae) has been recognized as an emerging agent of pneumonia as it has been identified in human samples via culture-based, molecular and serological techniques. Additionally, studies on animal models have shown that it fulfills the third and fourth Koch postulates to be assigned a pathogenic role. Due to the threat posed by it, multiple tools have been employed in the search for P. acanthamoebae. The methods utilized for its detection would be cell culture based approaches which involve both animal and amoebal cell culture and also molecular techniques that encompasses polymerase chain reaction (PCR), fluorescence in situ hybridization (FISH) and in situ hybridization (ISH). Additionally, immunohistochemistry (IHC) and serology based techniques such as direct and indirect immunofluorescence are also employed with the usage of Western blotting or immunoblotting as confirmatory procedures. This review attempts to describe the variety of techniques that are present in literature for the isolation and identification of P. acanthamoebae.

New developments in online OUR monitoring and its application to animal cell cultures.

The increasing demand for biopharmaceuticals produced in mammalian cells has driven the industry to enhance the productivity of bioprocesses through intensification of culture process. Fed-batch and perfusion culturing strategies are considered the most attractive choices, but the application of these processes requires the availability of reliable online measuring systems for the estimation of cell density and metabolic activity. This manuscript reviews the methods (and the devices used) for monitoring of the oxygen consumption, also known as oxygen uptake rate (OUR), since it is a straightforward parameter to estimate viable cell density and the physiological state of cells. Furthermore, as oxygen plays an important role in the cell metabolism, OUR has also been very useful to estimate nutrient consumption, especially the carbon (glucose and glutamine) and nitrogen (glutamine) sources. Three different methods for the measurement of OUR have been developed up to date, being the dynamic method the golden standard, even though DO and pH perturbations generated in the culture during each measurement. For this, many efforts have been focused in developing non-invasive methods, such as global mass balance or stationary liquid mass balance. The low oxygen consumption rates by the cells and the high accuracy required for oxygen concentration measurement in the gas streams (inlet and outlet) have limited the applicability of the global mass balance methodology in mammalian cell cultures. In contrast, stationary liquid mass balance has successfully been implemented showing very similar OUR profiles compared with those obtained with the dynamic method. The huge amount of studies published in the last years evidence that OUR have become a reliable alternative for the monitoring and control of high cell density culturing strategies with very high productivities.

Virucidal activity of Garcinia parvifolia leaf extracts in animal cell culture

BackgroundGarcinia species contain bioactive compounds such as flavonoids, xanthones, triterpernoids, and benzophenones with antibacterial, antifungal, anti-inflammatory, and antioxidant activities. In addition, many of these compounds show interesting biological properties such as anti-human immunodeficiency virus activity. Garcinia parvifolia is used in traditional medicine. Currently, the antiviral activity of G. parvifolia is not known.MethodsThis study was conducted to determine the effects of ethyl acetate (45 L Ea), ethanol (45 L Et), and hexane (45 L H) leaf extracts of G. parvifolia on the infectivity of pseudorabies virus (PrV) in Vero cells. The antiviral effects of the extracts were determined by cytopathic effect (CPE), inhibition, attachment, and virucidal assays.ResultsThe 50% cytotoxicity concentration (CC50) values obtained were 237.5, 555.0, and < 1.25 μg/mL for 45 L Ea, 45 L Et, and 45 L H, respectively. The 45 L Ea showed the greatest viral inhibition potency of 75% at 125 μg/mL. Both 45 L Ea and 45 l Et caused 100% residual viral inhibition at 250 μg/mL. The selectivity index values for 45 L Ea, 45 L Et, and 45 L H were 2.65, 1.75, and 0.10 showing that 45 L Ea had the greatest antiviral activity among the three extracts.ConclusionThis study showed that ethyl acetate is the best solvent to be used to obtain extract from G. parvifolia leaves with potent antiviral activities.

Pathophysiology and Mechanisms of Zika Virus Infection in the Nervous System.

In 2015, public awareness of Zika virus (ZIKV) rose in response to alarming statistics of infants with microcephaly being born to women who were infected with the virus during pregnancy, triggering global concern over these potentially devastating consequences. Although we have discovered a great deal about the genome and pathogenesis of this reemergent flavivirus since this recent outbreak, we still have much more to learn, including the nature of the virus-host interactions and mechanisms that determine its tropism and pathogenicity in the nervous system, which are in turn shaped by the continual evolution of the virus. Inevitably, we will find out more about the potential long-term effects of ZIKV exposure on the nervous system from ongoing longitudinal studies. Integrating clinical and epidemiological data with a wider range of animal and human cell culture models will be critical to understanding the pathogenetic mechanisms and developing more specific antiviral compounds and vaccines.

Targeting CD39 Toward Activated Platelets Reduces Systemic Inflammation and Improves Survival in Sepsis: A Preclinical Pilot Study.

Sepsis is associated with a systemic inflammatory reaction, which can result in a life-endangering organ dysfunction. Pro-inflammatory responses during sepsis are characterized by increased activation of leukocytes and platelets, formation of platelet-neutrophil aggregates, and cytokine production. Sequestration of platelet-neutrophil aggregates in the microvasculature contributes to tissue damage during sepsis. At present no effective therapeutic strategy to ameliorate these events is available. In this preclinical pilot study, a novel anti-inflammatory approach was evaluated, which targets nucleoside triphosphate hydrolase activity toward activated platelets via a recombinant fusion protein combining a single-chain antibody against activated glycoprotein IIb/IIIa and the extracellular domain of CD39 (targ-CD39). Experimental animal study and cell culture study. University-based experimental laboratory. Human dermal microvascular endothelial cells 1, human platelets and neutrophils, and C57BL/6NCrl mice. Platelet-leukocyte-endothelium interactions were evaluated under inflammatory conditions in vitro and in a murine lipopolysaccharide-induced sepsis model in vivo. The outcome of polymicrobial sepsis was evaluated in a murine cecal ligation and puncture model. To evaluate the anti-inflammatory potential of activated platelet targeted nucleoside triphosphate hydrolase activity, we employed a potato apyrase in vitro and in vivo, as well as targ-CD39 and as a control, nontarg-CD39 in vivo. Under conditions of sepsis, agents with nucleoside triphosphate hydrolase activity decreased platelet-leukocyte-endothelium interaction, transcription of pro-inflammatory cytokines, microvascular platelet-neutrophil aggregate sequestration, activation marker expression on platelets and neutrophils contained in these aggregates, leukocyte extravasation, and organ damage. Targ-CD39 had the strongest effect on these variables and retained hemostasis in contrast to nontarg-CD39 and potato apyrase. Most importantly, targ-CD39 improved survival in the cecal ligation and puncture model to a stronger extent then nontarg-CD39 and potato apyrase. Targeting nucleoside triphosphate hydrolase activity (CD39) toward activated platelets is a promising new treatment concept to decrease systemic inflammation and mortality of sepsis. This innovative therapeutic approach warrants further development toward clinical application.

Semi-perfusion cultures of suspension MDCK cells enable high cell concentrations and efficient influenza A virus production

Control and prevention of rapid influenza spread among humans depend on the availability of efficient and safe seasonal and pandemic vaccines, made primarily from inactivated influenza virus particles. Current influenza virus production processes rely heavily on embryonated chicken eggs or on cell culture as substrate for virus propagation. Today’s efforts towards process intensification in animal cell culture could innovate viral vaccine manufacturing using high-yield suspension cells in high cell density perfusion processes. In this work, we present a MDCK cell line adapted to grow as single cell suspension with a doubling time of less than 20 h, achieving cell concentrations over 1 × 107 cells/mL in batch mode. Influenza A virus titer obtained in batch infections were 3.6 log10(HAU/100 µL) for total- and 109 virions/mL for infectious virus particles (TCID50), respectively. In semi-perfusion mode concentrations up to 6 × 107 cells/mL, accumulated virus titer of 4.5 log10(HAU/100 µL) and infectious titer of almost 1010 virions/mL (TCID50) were possible. This exceeds results reported previously for cell culture-based influenza virus propagation by far and suggests perfusion cultures as the preferred method in viral vaccine manufacturing.

Heat-inactivated coelomic fluid of the earthworm Perionyx excavatus is a possible alternative source for fetal bovine serum in animal cell culture.

Fetal Bovine Serum (FBS) is used as a major supplement in culturing animal cells under in vitro conditions. Due to ethical concern, high cost, biosafety, and geographical as well as batchwise result variations, it is important to reduce or replace the use of FBS in animal cell culture. The major objective of this work is to evaluate the feasibility of heat-inactivated coelomic fluid (HI-CF) of the earthworm, Perionyx excavatus as a possible alternative for FBS in animal cell culture experiments. The coelomic fluid (CF) was extruded from the earthworm using electric shock method and used for the experiments. Electric shock method is a simple non-invasive technique, which has no harmful effect on earthworms. Mouse primary fibroblast and HeLa cell lines were used in this study. Among HI-CF, autoclaved CF and crude CF, the supplement of medium with HI-CF shows positive results. The processed HI-CF (90°C for 5 min) at 10% supplement in cell culture medium promote maximum cell growth but cells need the initial support of FBS for the attachment to the culture flask. Microscopic observation and immunofluorescence assay with actin and lamin A confirm that the cellular and molecular morphology of the cells is maintained intact. The HI-CF of earthworm, P. excavatus has shown better cellular viability when compared with FBS and making it possible as an alternative supplement to minimize the use of FBS.

Discovery of a Novel Connecting Link between Renin-Angiotensin System and Cancer in Barrett's Esophagus by Proteomic Screening.

The renin-angiotensin system (RAS) plays a central role in the regulation of homeostasis and blood pressure. This involves an important enzyme called angiotensin-converting enzyme that leads to the conversion of angiotensin I into angiotensin II. RAS has been reported to show association with inflammation, and in sporadic studies, with cancer. In particular, angiotensin II has been reported to be prevalent in the hypoxic microenvironment and associated with cancer signaling pathways. In a recent study, Bratlie etal. (Proteomics Clin. Appl. 2019, 4, 1800102) is shown to exploit 2D gel electrophoresis, and mass spectrometry (MS) to identify differentially expressed proteins by comparing low-grade dysplasia in Barrett's Esophagus (BE) following administration of agents that interfere with RAS, that is, enalapril and candesartan, and identified specific modulation of HSP60, PDIA3, and PPA1. Though 2D gel coupled with MS is a commonly-used tool for studying proteomes, it still has limitations in terms of a comprehensive analysis due to lack of absolute quantitation in a high-throughput manner. Despite technical limitations and the small size of the study, preliminary data emerging from the investigation show interference caused by clinically approved RAS inhibitors resulting in alteration of molecular markers associated with tumorigenicity. The authors propose potential factors that may influence the progression of the disease. However, these are conspicuous changes in high-abundance proteins only. Therefore, there is a need to carry out detailed experimental studies either using an in vitro labeling technique (isobaric labeling for relative and absolute quantitation) for tissues or an in vivo labeling technique (stable isotope labeling in animal cell culture) coupled with LC-MS/MS to identify differentially-regulated proteins to delineate the role of RAS in BE.

Bumped kinase inhibitor 1369 is effective against Cystoisospora suis in vivo and in vitro

Cystoisosporosis is a leading diarrheal disease in suckling piglets. With the confirmation of resistance against the only available drug toltrazuril, there is a substantial need for novel therapeutics to combat the infection and its negative effects on animal health. In closely related apicomplexan species, bumped kinase inhibitors (BKIs) targeting calcium-dependent protein kinase 1 (CDPK1) were shown to be effective in inhibiting host-cell invasion and parasite growth. Therefore, the gene coding for Cystoisospora suis CDPK1 (CsCDPK1) was identified and cloned to investigate activity and thermal stabilization of the recombinant CsCDPK1 enzyme by BKI 1369. In this comprehensive study, the efficacy, safety and pharmacokinetics of BKI 1369 in piglets experimentally infected with Cystoisospora suis (toltrazuril-sensitive, Wien-I and toltrazuril-resistant, Holland-I strains) were determined in vivo and in vitro using an established animal infection model and cell culture, respectively. BKI 1369 inhibited merozoite proliferation in intestinal porcine epithelial cells-1 (IPEC-1) by at least 50% at a concentration of 40 nM, and proliferation was almost completely inhibited (>95%) at 200 nM. Nonetheless, exposure of infected cultures to 200 nM BKI 1369 for five days did not induce structural alterations in surviving merozoites as confirmed by transmission electron microscopy. Five-day treatment with BKI 1369 (10 mg/kg BW twice a day) effectively suppressed oocyst excretion and diarrhea and improved body weight gains in treated piglets without obvious side effects for both toltrazuril-sensitive, Wien-I and resistant, Holland-I C. suis strains. The plasma concentration of BKI 1369 in piglets increased to 11.7 μM during treatment, suggesting constant drug accumulation and exposure of parasites to the drug. Therefore, oral applications of BKI 1369 could potentially be a therapeutic alternative against porcine cystoisosporosis. For use in pigs, future studies on BKI 1369 should be directed towards ease of drug handling and minimizing treatment frequencies.

CYTOTOXICITY STUDY OF ULTRASMALL PHOSPHONIUM GOLD NANOPARTICLES USING PLANT AND ANIMAL CELL CULTURES

Abstract—The biocompatibility of ultrasmall colloidal gold nanoparticles seems an important problem due to their expanding range of biomedical and technical applications every year. According to most studies, this type of nanoparticle is toxic to living organisms. However, it is not clear which particular component of the colloidal system exhibits toxicity: whether it is associated with the particles themselves or with the dispersion medium. Also, the mechanism of the toxic effect is not clear. Solving this problem goes hand in hand with identifying the source of toxicity in the preparation of ultrasmall phosphonium gold nanoparticles, obtained by the Duff method using the following cell test systems: microalgae cultures of Dunaliella salina and animal cell cultures of the HeLa and Vero lines. Nanoparticles washed three times of the medium were not toxic to animal cells and were slightly toxic to D. salina. It was found that the toxicity of the preparation of ultrasmall phosphonium gold nanoparticles is controlled by the toxicity of the dispersion medium; it can be assumed that complex-ionic forms of gold are the main source of toxicity in the dispersion medium.

Three-dimensional decellularized amnion membrane scaffold as a novel tool for cancer research; cell behavior, drug resistance and cancer stem cell content

BackgroundCancer is the second leading cause of human death. Therefore, comprehensive research and the appropriate tools are needed in this field. Animal models and cell culture studies are the most important preclinical tools in cancer research. In 2D cell culture models, cells are forced to grow in a 2D environment, which differs from their natural physiology. Recently, 3D cell culture models were developed to fill the gap between 2D cell culture and animal models. Materials and methodsHuman amniotic membranes were obtained, decellularized, characterized and used as a natural 3D scaffold to investigate cancer cell behavior in 2D compared to 3D conditions. Time-lapse imaging of cells was used, and cell proliferation, velocity and migration were evaluated. Cisplatin was applied in 2D and 3D conditions, followed by evaluation of viability, apoptosis and cancer stem cell proteins by flow cytometry and western blot analysis. ResultsThe results showed that in the decellularized amnion membrane (DAM) scaffold most cells did not spread and remain rounded and then penetrated into the scaffold with no cytotoxicity. Significant differences in migration, velocity, morphology and proliferation of cancer cells were observed between the 3D DAM scaffold and 2D model. Furthermore, the cells in the 3D DAM scaffold showed much more resistance to apoptosis and higher CSC content. ConclusionIn conclusion, considering the effect of the 3D DAM scaffold in cell behavior, apoptosis resistance and CSC content as well as the short processing time for decellularizing the AM, it appears that the 3D DAM scaffold offers an appropriate tool for in vitro cancer research.

Multiscale modeling of influenza A virus replication in cell cultures predicts infection dynamics for highly different infection conditions.

Influenza A viruses (IAV) are commonly used to infect animal cell cultures for research purposes and vaccine production. Their replication is influenced strongly by the multiplicity of infection (MOI), which ranges over several orders of magnitude depending on the respective application. So far, mathematical models of IAV replication have paid little attention to the impact of the MOI on infection dynamics and virus yields. To address this issue, we extended an existing model of IAV replication in adherent MDCK cells with kinetics that explicitly consider the time point of cell infection. This modification does not only enable the fitting of high MOI measurements, but also the successful prediction of viral release dynamics of low MOI experiments using the same set of parameters. Furthermore, this model allows the investigation of defective interfering particle (DIP) propagation in different MOI regimes. The key difference between high and low MOI conditions is the percentage of infectious virions among the total virus particle release. Simulation studies show that DIP interference at a high MOI is determined exclusively by the DIP content of the seed virus while, in low MOI conditions, it is predominantly controlled by the de novo generation of DIPs. Overall, the extended model provides an ideal framework for the prediction and optimization of cell culture-derived IAV manufacturing and the production of DIPs for therapeutic use.

An LDHa single allele CHO cell mutant exhibits altered metabolic state and enhanced culture performance

AbstractBACKGROUNDReducing lactate production in animal cell cultures has been reported to improve cell culture performance and productivity of recombinant protein. A novel genome editing tool, CRISPR/Cas, has been used widely to induce double‐strand breaks in the genome and introduce targeted mutations efficiently.RESULTSIn the present work, we used a publicly available human‐codon optimized CRISPR/Cas system to introduce mutations in one of the LDHa gene copies to obtain an LDHa single‐allele knockout Chinese hamster ovary (CHO) cell clone to analyze its effect over cell metabolism. Fed‐batch cultures were conducted in order to evaluate the culture performance of mutant cells. Results show that cell growth was reduced and metabolism was modified by the LDHa single‐allele knockout, whereas specific protein rate and volumetric production were greatly enhanced. Additionally, the first in‐depth analysis of the metabolic effects of LDHa single allele knockout is presented.CONCLUSIONBy using CRISPR/Cas we were able to knock down LDHa, showing improved culture productivity. Metabolic flux analysis revealed higher fluxes in the TCA cycle in the mutant cell than in the parental cell, indicating that LDHa mutant cells have an enhanced energy metabolism, which can be attributed mainly to a higher catabolism of amino acids. © 2018 Society of Chemical Industry

Control of IgG glycosylation by in situ and real-time estimation of specific growth rate of CHO cells cultured in bioreactor.

The cell-specific growth rate (µ) is a critical process parameter for antibody production processes performed by animal cell cultures, as it describes the cell growth and reflects the cell physiological state. When there are changes in these parameters, which are indicated by variations of µ, the synthesis and the quality of antibodies are often affected. Therefore, it is essential to monitor and control the variations of µto assure the antibody production and achieve high product quality. In this study, a novel approach for on-line estimation of µ was developed based on the process analytical technology initiative by using an in situ dielectric spectroscopy. Critical moments, such as significant µ decreases, were successfully detected by this method, in association with changes in cell physiology as well as with an accumulation of nonglycosylated antibodies. Thus, this method was used to perform medium renewals at the appropriate time points, maintaining the values of µ close to its maximum. Using this method, we demonstrated that the physiological state of cells remained stable, the quantity and the glycosylation quality of antibodies were assured at the same time, leading to better process performances compared with the reference feed-harvest cell cultures carried out by using off-line nutrient measurements.

The antileukemic effects of saffron (Crocus sativus L.) and its related molecular targets: A mini review.

Saffron (Crocus sativus L.), and its main constituents, crocin, and crocetin have shown promising effects as an antileukemic agent in animal models and cell culture systems. Saffron retards the growth of cancer cells via inhibiting nucleic acid synthesis and enhancing antioxidative system. It can induce apoptosis and chemosensitivity via inhibiting multidrug resistance proteins. Saffron also induces differentiation pathways via inhibiting promyelocytic leukemia/retinoic acid receptor-α, histone deacetylase1, and tyrosyl DNA phosphodiesterase-1 as well. The present review highlights the most recent findings on the antileukemic effects of saffron and its underlying molecular targets. The emerging evidence suggests that saffron has a selective toxicity effect against leukemic cells while is safe for the normal cells.

Analysis of the immunoglobulin G (IgG) secretion efficiency in recombinant Chinese hamster ovary (CHO) cells by using Citrine-fusion IgG.

Biopharmaceuticals represented by immunoglobulin G (IgG) are produced by the cultivation of recombinant animal cells, especially Chinese hamster ovary (CHO) cells. It is thought that the intracellular secretion process of IgG is a bottleneck in the production of biopharmaceuticals. Many studies on the regulation of endogenous secretory protein expression levels have shown improved productivity. However, these strategies have not universally improved the productivity of various proteins. A more rational and efficient establishment of high producer cells is required based on an understanding of the secretory processes in IgG producing CHO cells. In this study, a CHO cell line producing humanized IgG1, which was genetically fused with fluorescent proteins, was established to directly analyze intracellular secretion. The relationship between the amount of intracellular and secreted IgG was analyzed at the single cell level by an automated single-cell analysis and isolation system equipped with dual color fluorescent filters. The amounts of intracellular and secreted IgG showed a weak positive correlation. The amount of secreted IgG analyzed by the system showed a weak negative linear correlation with the specific growth of isolated clones. An immunofluorescent microscopy study showed that the established clones could be used to analyze the intracellular secretion bottleneck. This is the first study to report the use of fluorescent protein fusion IgG as a tool to analyze the secretion of recombinant CHO cells.

New approaches to improve efficacy and reduce side effects of phototherapy for neonatal hyperbilirubinemia using led and laser sources

The article discusses the new opportunities for improving technology of phototherapy for neonatal jaundice using LED and laser sources in order to reduce the level of bilirubin in the blood of infants. It has been shown that several types of LEDs with emission spectrum corresponding to the long-wavelength slope of bilirubin absorption band can be used for therapeutic purposes. At the same time, the efficacy of phototherapy depend on not only from the position of the maximum in emission spectrum of the LEDs within bilirubin absorption band, but also on the spectral width of acting radiation. Based on the study of regularities of infl of optical radiation on the survival of animal cells in culture sensitized with bilirubin it has been concluded that the damaging eff cts are signifi ly reduced upon transition from quasi-monochromatic LED sources to lasers, emitting in the same spectral range. It allows to consider the modern semiconductor lasers emitting in blue and blue-green spectral regions as promising sources for development of phototherapeutic equipment for treating neonatal hyperbilirubinemia.

SU21 - INTEGRATED ANALYSIS OF MICROARRAY AND PROTEOMICS DATA BY HIPPOCAMPAL RAT NEURONS AND HUMAN IPS NEURONS TREATED WITH DIFFERENT DOSES OF LITHIUM

Background Lithium salts have a well-established role in the treatment of psychiatric disorders as major affective disorders and more recently Alzheimer disease. Experimental and clinical studies have provided evidence that lithium may exert neuroprotective effects at therapeutically and sub therapeutically doses. In animal model and cell culture models, lithium has been shown to increase neuronal viability through a combination of mechanisms that includes the inhibition of apoptosis, regulation of autophagy, increased mitochondrial function, and synthesis of neurotrophic factors. Objective: The Objective of the study is to compare biological process and pathways enriched in hippocampal neurons treated with different doses of lithium (0.02 mM, 0.2 mM and 2 mM) integrating genes and proteins differentially expressed. Methods Primary cultures of hippocampal neurons; Assessment of cell viability; Experimental design considerations, Microarray; LC-MS analyses; Data Analysis; Neural stem cell differentiation in Random differentiation; String Database and Functional enrichment analysis. Results The results showed that growing gene networks using differentially expressed genes and proteins as seeds alterations in mitochondria, ribosome and proteasome were observed in all lithium treatment doses. But also more specific biological process such as mitochondria respiratory chain, energy production (glucose and ATP) and apoptosis biological functions were enriched in each dose treatment. Discussion Given all the analyzes we can see that there is a dissociation dose effect of the treatment with lithium, the dose of 0.02Mm modified genes of degradation pathway proteins. The dose of 0.2Mm altered pathways of energy metabolism and the dose of 2Mm change related to nuclear and nuclear membrane. we can understand the specific function of each dose of the treatment.