Cellular Protein Content Research Articles

Phylogenetic Analysis and Biological Evaluation of Marine Endophytic Fungi Derived from Red Sea Sponge Hyrtios erectus.

Forty-four endophytic fungal isolates obtained from marine sponge, Hyrtios erectus, were evaluated and screened for their hydrolase activities. Most of the isolates were found to be prolific producers of hydrolytic enzymes. Only 11 isolates exhibited maximum cellular contents of lipids, rhamnolipids, and protein in the fungal isolates under the isolation numbers MERVA5, MERVA22, MERVA25, MERVA29, MERVA32, MERVA34, MERV36, MERVA39, MERVA42, MERVA43, and MERVA44. These isolate extracts exhibit the highest reducing activities against carbohydrate-metabolizing enzymes including α-amylase, α-glucosidase, β-glucosidase, β-glucuronidase, and tyrosinase. Consequently, based on morphological and cultural criteria, as well as sequence information and phylogenetic analysis, these isolates could be identified and designated as Penicillium brevicombactum MERVA5, Arthrinium arundinis MERVA22, Diaporthe rudis MERVA25, Aspergillus versicolor MERVA29, Auxarthron alboluteum MERVA32, Dothiorella sarmentorum MERVA34, Lophiostoma sp. MERVA36, Fusarium oxysporum MERVA39, Penicillium chrysogenum MERVA42, Penicillium polonicum MERVA43, and Trichoderma harzianum MERVA44. The endophytic fungal species, D. rudis MERVA25, P. polonicum MERVA43, Lophiostoma sp. MERVA36, A. alboluteum MERVA32, T. harzianum MERVA44, F. oxysporum MERVA39, A. versicolor MERVA29, and P. chrysogenum MERVA42 extracts, showed significant hepatitis C virus (HCV) inhibition. Moreover, D. sarmentorum MERVA34, P. polonicum MERVA43, and T. harzianum MERVA44 extracts have the highest antitumor activity against human hepatocellular carcinoma cells (HepG2).

Lytic and Chemotactic Features of the Plaque-Forming Bacterium KD531 on Phaeodactylum tricornutum.

Phaeodactylum tricornutum is a dominant bloom forming species and potential biofuel feedstock. To control P. tricornutum bloom or to release lipids from P. tricornutum, we previously screened and identified the lytic bacterium Labrenzia sp. KD531 toward P. tricornutum. In the present study, we evaluated the lytic activity of Labrenzia sp. KD531 on microalgae and investigated its lytic mechanism. The results indicated that the lytic activity of KD531 was temperature- and pH-dependent, but light-independent. In addition to P. tricornutum, KD531 also showed lytic activity against other algal species, especially green algae. A quantitative analysis of algal cellular protein, carbohydrate and lipid content together with measurements of dry weight after exposure to bacteria-infected algal lysate indicated that the bacterium KD531 influenced the algal biomass by disrupting the algal cells. Both chemotactic analysis and microscopic observations of subsamples from different regions of formed plaques showed that KD531 could move toward and then directly contact algal cells. Direct contact between P. tricornutum and KD531 cells was essential for the lytic process.

Insulin-like Effects of Mt. Fuji Subsoil Water which Conta ins Vanadium on Cultured Cells: Insight from Japan

Background: It was reported that vanadium compounds possess insulin-like activity, although suitable vanadium compounds have not yet developed as anti-diabetes mellitus drugs. Objective: As Mt. Fuji subsoil water contains high levels of vanadium, this study aimed to assess whether it showed insulin-like effects. Method: Insulin-like effects of vanadium ions have been investigated in cultured cells, using rat fibroblasts (Py-3Y1-S2), human esophageal cancer cells (TE-13), and other established cell lines. To evaluate insulin-like effects, glucose consumption in the cultured media and cell growth based on cellular protein content were measured, and cytotoxic effects on cells were assessed. Results: The vanadium concentration in the commercially available Mt. Fuji subsoil water ranged from 56–144 ppb, and the highest concentration (144.2 ± 1.2 ppb) was observed in Healthy Vana Water. The vanadium concentration in other water samples derived from wells, ravine rivers, and cities ranged from 0.5 to 27 ppb, and a comparatively high vanadium concentration (27.1 ± 0.0 ppb) was observed in the well water at Kayayama-Machi, which is close to Mt. Fuji. The largest insulin-like effect on glucose consumption was observed in Healthy Vana Water among the various water samples tested. The effects of vanadium at low concentrations of 200-500 µg/l seemed to accelerate cell growth, while high concentrations of vanadium (> 500 µg/l) had a cytotoxic effect. Conclusion: Mt. Fuji subsoil water possesses higher insulin-like activity than other water samples derived from wells, ravine rivers, and cities, suggesting a potential future role in diabetes mellitus therapy. Keywords: Insulin-like effects, diabetes, vanadium, Mt. Fuji subsoil water, glucose consumption, cell growth, cytotoxicity.

Radiation Mitigating Properties of Intranasally Administered KL4Surfactant in a Murine Model of Radiation-Induced Lung Damage

The threat of exposure to ionizing radiation from a nuclear reactor accident or deliberate terrorist actions is a significant public health concern. The lung is particularly susceptible to radiation-induced injury from external sources or inhalation of radioactive particles from radioactive fallout. Radiation-induced lung disease can manifest with an acute radiation pneumonitis and/or delayed effects leading to pulmonary fibrosis. As prior warning of radiation exposure is unlikely, medical countermeasures (MCMs) to mitigate radiation-induced lung disease that can be given in mass-casualty situations many hours or days postirradiation are needed to prevent both early and late lung damage. In this study, KL4 surfactant (lucinactant) was evaluated as a radiation mitigator in a well-characterized mouse model of targeted thoracic radiation exposure, for its effect on both early (several weeks) and late (18 weeks) lung damage. Here, 120 mg/kg total phospholipid of KL4 surfactant was administered twice daily intranasally, (enabling intrapulmonary inhalation of drug) to C57BL/6 mice 24 h after a single 13.5 Gy dose of thoracic irradiation (LD50 dose). Both early and chronic phase (2 and 4 weeks and 18 weeks postirradiation, respectively) assessments were performed. Mice were evaluated for evidence of reduced arterial blood oxygenation and early and chronic lung and systemic inflammation, lung fibrosis and oxidative stress. Analysis was done by performing lung function/respiration dynamics and measuring cellular protein content of bronchoalveolar lavage fluid (BALF), and levels of cytokines, 8-iso-prostaglandin F2α, hydroxyproline in lung and plasma, along with evaluating lung histology. The results of this study showed that intranasal delivery of KL4 surfactant was able to preserve lung function as evidenced by adequate arterial oxygen saturation and reduced lung inflammation and oxidative stress; total white count and absolute neutrophil count was decreased in BALF, as were plasma pro-inflammatory cytokine levels and biomarker of oxidative stress. KL4 surfactant is a promising MCM for mitigation of lung tissue damage after targeted, thoracic irradiation and has the potential to be developed as a broad-spectrum, multi-use MCM against chemical, biological, radiological or nuclear threat agents with potential to cause lung injury.

Antioxidant, anti-inflammatory, antiproliferative and antimycobacterial activities of the essential oil of Psidium guineense Sw. and spathulenol

Ethnopharmacological relevanceLeaves from Psidium guineense Sw. are used in popular medicine for the treatment of inflammatory disease. However, there is no scientific evidence demonstrating this activity. Aim of the studyTo evaluate the antioxidant, anti-inflammatory, antiproliferative and antimycobacterial activities of the essential oil of P. guineense and spathulenol (a major constituent). The study was conducted in part to provide evidence supporting the ethnobotanical use of the leaves of this species. Material and methodsThe essential oil (EOPG) was extracted from the leaves of P. guineense by hydrodistillation and analysed by gas chromatography-mass spectrometry (GC–MS). The major compound, spathulenol (PG-1), was isolated in a chromatographic column and characterized by nuclear magnetic resonance (NMR). EOPG and PG-1 were evaluated in vitro for antioxidant activity by DPPH, ABTS and MDA methods; anti-inflammatory potential was assessed using two models, including pleurisy and oedema, in mice. The impact of EOPG and PG-1 on cell proliferation was determined via spectrophotometric quantification of the cellular protein content using a sulforhodamine B assay, and anti-Mycobacterium tuberculosis activity was determined using the REMA method. ResultsA total of 38 components were identified from the EOPG, with the sesquiterpenic alcohol spathulenol (PG-1) (80.7%) being the major constituent. EOPG and PG-1 exhibited the highest antioxidant activities in the DPPH and MDA system compared with reference standard, with IC50 values ranging from 26.13 to 85.60μg/mL. Oral administration of EOPG and PG-1 showed significant inhibition in the Cg-induced mice paw oedema and pleurisy model. The EOPG (GI50 = 0.89μg/mL) and PG-1 (GI50 = 49.30μg/mL) were particularly effective against the ovarian cancer cell line. Both showed moderate antimycobacterial activity. ConclusionFor the first time, this study demonstrated the antioxidant, anti-inflammatory, antiproliferative and antimycobacterial properties of the essential oil of P. guineense (leaves were collected in Dourados-MS) and spathulenol, collaborating the etnhopharmacologycal use of this plant due to its an anti-inflammatory effect.

MiR‑21 promotes α‑SMA and collagen I expression in hepatic stellate cells via the Smad7 signaling pathway.

The aim of the present study was to investigate the role of microRNA (miR)‑21 in regulating collagenI and Smad7 expression in activated rat hepatic stellate cells (HSCs). Rat HSCs were isolated by single‑step density gradient centrifugation with Nycodenz. Cellular content of miR‑21, SMAD7, α‑smooth muscle actin (α‑SMA), collagen type I alpha 1 (COLLA1) and COLL alpha 2 (A2) mRNA was examined by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR), and cellular content of Smad7 and α‑SMA protein was detected by western blotting. Binding of miR‑21 to the 3'‑untranslated region (UTR) of Smad7 was verified by dual‑luciferase assay. The authors observed that, in activated HSCs, expression of miR‑21 was significantly increased in a time‑dependent manner, while expression of Smad7 mRNA and protein was significantly reduced. In addition, miR‑21 mimics significantly enhanced cellular α‑SMA mRNA and protein content, while miR‑21 inhibitor significantly reduced α‑SMA mRNA and protein levels. Similarly, cellular content of COLLA1 and COLLA2 mRNA was significantly elevated by miR‑21 mimics, but reduced by miR‑21 inhibitor, in activated HSCs. Moreover, cellular content of Smad7 mRNA and protein was significantly reduced by miR‑21 mimics, but significantly increased by miR‑21 inhibitor. Furthermore, miR‑21 mimics activated firefly luciferase in HEK293 cells transfected with the wild type 3'‑UTR of Smad7. miR‑21 regulates expression of α‑SMA and collagen I in activated rat HSCs by directly targeting Smad7.

Effect of bacteria on growth and biochemical composition of two benthic diatoms Halamphora coffeaeformis and Entomoneis paludosa

Benthic diatoms are the dominant microalgae in intertidal mudflats and are in constant interaction with their surrounding bacteria. This study was designed to investigate the effect of bacteria on growth, biomass, elemental (C & N) and biochemical composition, and extracellular polymeric substances (EPS) excretion by two marine benthic diatoms, Halamphora coffeaeformis and Entomoneis paludosa. The experiments were conducted on diatom cultures previously exposed or not to antibiotics. The treatment with antibiotics caused a decrease of bacterial abundance from 24 to fewer than 1 bacteria per algal cell. In non-treated cultures of E. paludosa and H. coffeaeformis, the bacteria phylogenetic affiliation was equally distributed between Bacteroidetes (Flavobacteriia) and Proteobacteria (alpha- and gammaproteobacteria). After treatment with antibiotics, the residual bacterial community was ~37% Flavobacteriia (Winogragskyella genus), 34% for the alphaproteobacteria (mainly Roseibacterium sp and Antarctobacter sp.) and 29% for the gammaproteobacteria (mainly Methylophaga sp. and Stenotrophomonas sp.). Growth of H. coffeaeformis and E. paludosa in non-treated cultures was enhanced by the abundance of the associated bacteria, with mean growth rate of 1day−1 compared to 0.7 for antibiotic treated cultures. In E. paludosa, maximal cell abundance was higher in the presence of bacteria while the final carbon biomass did not vary, but in H. coffeaeformis maximal cell abundance did not vary significantly while final carbon biomass was higher in the presence of bacteria. By contrast, for both diatoms, cellular content of protein and lipids decrease significantly, as did extracellular carbon (EPS fraction) in the presence of bacteria. However, only a minor effect was observed on cellular carbohydrates, C/N ratio, and pigments (Chl a). Diatoms carbon fluxes towards the main biochemical components were also modified, with the protein carbon fraction significantly lower relative to other carbon compounds in the presence of high bacterial biomass. These results showed the complex interactions between diatoms and their associated bacteria. Promotion of diatoms growth by the presence of bacteria appears linked to change in microalgae biochemical composition that will modify the biofilm. Our results might help understanding the regulation of benthic biota in mudflat ecosystems.

Identification and Metabolite Profiling of Chemical Activators of Lipid Accumulation in Green Algae.

Microalgae are proposed as feedstock organisms useful for producing biofuels and coproducts. However, several limitations must be overcome before algae-based production is economically feasible. Among these is the ability to induce lipid accumulation and storage without affecting biomass yield. To overcome this barrier, a chemical genetics approach was employed in which 43,783 compounds were screened against Chlamydomonas reinhardtii, and 243 compounds were identified that increase triacylglyceride (TAG) accumulation without terminating growth. Identified compounds were classified by structural similarity, and 15 were selected for secondary analyses addressing impacts on growth fitness, photosynthetic pigments, and total cellular protein and starch concentrations. TAG accumulation was verified using gas chromatography-mass spectrometry quantification of total fatty acids, and targeted TAG and galactolipid measurements were performed using liquid chromatography-multiple reaction monitoring/mass spectrometry. These results demonstrated that TAG accumulation does not necessarily proceed at the expense of galactolipid. Untargeted metabolite profiling provided important insights into pathway shifts due to five different compound treatments and verified the anabolic state of the cells with regard to the oxidative pentose phosphate pathway, Calvin cycle, tricarboxylic acid cycle, and amino acid biosynthetic pathways. Metabolite patterns were distinct from nitrogen starvation and other abiotic stresses commonly used to induce oil accumulation in algae. The efficacy of these compounds also was demonstrated in three other algal species. These lipid-inducing compounds offer a valuable set of tools for delving into the biochemical mechanisms of lipid accumulation in algae and a direct means to improve algal oil content independent of the severe growth limitations associated with nutrient deprivation.

Polymer nanoassemblies with hydrophobic pendant groups in the core induce false positive siRNA transfection in luciferase reporter assays

Poly(ethylene glycol)-conjugated polyethylenimine (PEG-PEI) is a widely studied cationic polymer used to develop non-viral vectors for siRNA therapy of genetic disorders including cancer. Cell lines stably expressing luciferase reporter protein typically evaluate the transfection efficacy of siRNA/PEG-PEI complexes, however recent findings revealed that PEG-PEI can reduce luciferase expression independent of siRNA. This study elucidates a cause of the false positive effect in luciferase assays by using polymer nanoassemblies (PNAs) made from PEG, PEI, poly-(l-lysine) (PLL), palmitate (PAL), and deoxycholate (DOC): PEG-PEI (2P), PEG-PEI-PAL (3P), PEG-PLL (2P’), PEG-PLL-PAL (3P’), and PEG-PEI-DOC (2PD). In vitro transfection and western blot assays of luciferase using a colorectal cancer cell line expressing luciferase (HT29/LUC) concluded that 2P and 2P’ caused no luciferase expression reduction while hydrophobically modified PNAs induced a 35–50% reduction (3P’<2PD<3P). Although cell viability remained stagnant, 3P triggered cellular stress responses including increased membrane porosity and decreased ATP and cellular protein concentrations. Raman spectroscopy suggested that hydrophobic groups influence PNA conformation changes, which may have caused over-ubiquitination and degradation of luciferase in the cells. These results indicate that hydrophobically modified PEG-PEI induces cellular distress causing over-ubiquitination of the luciferase protein, producing false positive siRNA transfection in the luciferase assay.

Paraffin embedded tissues: sterile or contaminated material?

IntroductionFormalin‐fixed and paraffin‐embedded tissue is an invaluable resource in research. Formaldehyde fixation has been the method of choice for tissue preservation in pathology for more than a century. Beside tissue structure, the cellular contents of proteins and nucleic acids are preserved by the cross‐linking between and within proteins in the tissue. This gives a tissue sample resilience to long‐term storage, usually at ambient room temperature. When any kind of tissue is removed, it is sent to a pathology lab for processing, examination, and diagnosis. This material is processed by a very well known technique, that comprises inclusion in paraffin. Such method allows preparation of slides to undergo histological analysis. After diagnosis is done, the remaining paraffin blocks are stored almost indefinitely.ObjectivesThis paper aims to investigate whether metabolically active bacteria could be detected within tonsils and lymphnodes removed for different reasons, and stored after proper histological examination in paraffin blocks. Moreover, we wanted to know if there would still be bacteria growing in the preserved tissues, as remaining microbiote or new bacterial contamination. At last, if these bacteria are present, could it affect the tissue structure.Material and methodsTwenty three paraffin blocks containing tissues from different lymphatic organs were sellected: nasopharyngeal tonsils, palatine tonsils and lymph nodes. The hole blocks were imprinted into agar‐cellulose media, and thin slices of the blocks were made and put in liquid culture tubes. The plates and culture tubes were then incubated overnight at an incubation temperature of 37 °C (98.6 °F). The blocks were then cleaned of surface paraffin and then the tissue inside was carved to obtain material for slides. Slides containing samples of each one of the specimens were submitted to Zieh‐Neelsen coloring, in order to investigate the presence of acid‐alcohol resistant bacilli (AARB).ResultsTwo palatine tonsil imprints were positive for Negative‐coagulase Staphylococcus, the same happened with 5 Nasopharyngeal tonsil imprints and 2 Lymph node imprints. All culture plates showed only one bacterial colony localized on the border of the imprinted block. All liquid culture tubes were negative for bacterial growth. All of the slides submitted to Zieh‐Neelsen coloring were negative to (AARB).DiscussionNegative‐coagulase Staphylococci may be present on the skin of the hands. Tissue embedded paraffin blocks are routinely and freely handled without use of latex gloves, so we can assume that probably, S. epidermidis was the colonizer, and it is possible to infer that it was caused by handling contamination of the surface of the blocks. Paraffin embedded blocks seem to remain a safe and easy way for tissue sample storage, specially over time, that ranged from 3 to 15 years in this study.ConclusionsOnly handling bacterial contamination could be detected within tonsils and lymph nodes stored in paraffin blocks. Paraffin embedded blocks seem to remain a safe and easy way for tissue sample storage.

Ugonin M, a Helminthostachys zeylanica Constituent, Prevents LPS-Induced Acute Lung Injury through TLR4-Mediated MAPK and NF-κB Signaling Pathways.

Helminthostachys zeylanica (L.) Hook. is plant that has been used in traditional Chinese medicine for centuries for the treatment of inflammation, fever, pneumonia, and various disorders. The aims of the present study are to figure out the possible effectiveness of the component Ugonin M, a unique flavonoid isolated from H. zeylanica, and to elucidate the mechanism(s) by which it works in the LPS-induced ALI model. In this study, Ugonin M not only inhibited the production of pro-inflammatory mediators such as NO, TNF-α, IL-1β, and IL-6, as well as infiltrated cellular counts and protein content in the bronchoalveolar lavage fluid (BALF) of lipopolysaccharides (LPS)-induced acute lung injury (ALI) mice, but also ameliorated the severity of pulmonary edemas through the score of a histological examination and the ratio of wet to dry weight of lung. Moreover, Ugonin M was observed to significantly suppress LPS-stimulated protein levels of iNOS and COX-2. In addition, we found that Ugonin M not only obviously suppressed NF-κB and MAPK activation via the degradation of NF-κB and IκB-α as well as ERK and p38MAPK active phosphorylation but also inhibited the protein expression level of TLR4. Further, Ugonin M treatment also suppressed the protein levels of MPO and enhanced the protein expressions of HO-1 and antioxidant enzymes (SOD, GPx, and CAT) in lung tissue of LPS-induced ALI mice. It is anticipated that through our findings, there is strong evidence that Ugonin M may exert a potential effect against LPS-induced ALI mice. Hence, Ugonin M could be one of the major effective components of H. zeylanica in the treatment of inflammatory disorders.

Flocculation in industrial strains ofSaccharomyces cerevisiae: role of cell wall polysaccharides and lectin-like receptors

Yeast flocculation is the reversible aggregation of yeast cells promoted by the interaction between lectin-like protein receptors with mannose side chains on adjacent cell walls. Flocculation is governed by several physiological factors, including the type of nutrient sugar available to yeast. We grew four industrial strains of Saccharomyces cerevisiae, representing applications in the brewing, winemaking and bioethanol sectors, to late stationary phase and quantified the cellular content of mannans, glucans and lectin-like proteins on yeast cell surfaces. Results indicated that brewing and champagne strains showed moderate to high flocculation ability when grown with glucose, fructose, maltose or galactose, whereas winemaking and fuel alcohol strains only showed moderate flocculation when grown on maltose and galactose. All yeast strains studied were weakly flocculent when grown on mannose. With regard to lectin-like receptors, their number played a more important role in governing yeast flocculation than the mannan and glucan contents in yeast cell walls. We conclude that all the industrial strains of S. cerevisiae belonged to New-Flo type on the basis of their flocculation behaviour observed when cultured on different sugars. Quantification of yeast cell wall polysaccharides and receptor sites indicates that mannan and glucan levels remain almost constant, irrespective of the strain under investigation. The main difference in flocculation characteristics in industrial yeast strains appears to be due to variations in concentrations of lectin-like cell surface receptors. Our findings may benefit brewers, winemakers and other yeast-based technologies in design of media to prevent premature flocculation during fermentation. Copyright © 2017 The Institute of Brewing & Distilling

Effect of O-chitosan nanoparticles on the development and membrane permeability of Verticillium dahliae

Verticillium dahliae, which causes wilting in over 300 woody and herbaceous plant species, is a representative of fungal plant diseases for which effective controls are still needed. In this study, the antifungal action of oleoyl-chitosan nanoparticles was investigated against V. dahliae. Media containing oleoyl-chitosan nanoparticles dramatically decreased the mycelium growth. The highest antifungal indexes were observed on media amended with 2mg/mL nanoparticles. Optical microscopy showed that spore germination and hyphae morphology were affected. Scanning electron microscopy and transmission electron microscopy demonstrated degenerative alterations including crumpled hyphae and spores, thickened cell walls, disappeared membranous organelles, massive vacuolation of the cytoplasm, and cell wall-plasmalemma separation. Fluorescence microscopy showed that nanoparticles were internalized by fungal cells. The sharp increase in the release of intracellular components and decrease of total cellular protein concentration demonstrated damaged cell membranes. Overall, the results indicate that oleoyl-chitosan nanoparticles have the potential to control phytopathogens in agriculture.

Compositional profiles of Rhodosporidium toruloides cells under nutrient limitation.

Lipid production by the red yeast Rhodosporidium toruloides was explored under nutrient limitation. To determine the compositional profiles of R. toruloides cells, samples were prepared using a continuous cultivation process under nutrient limitation and analyzed via several methods, including Fourier transform infrared spectroscopy and elemental analysis. Under nitrogen limitation, as the dilution rate increased, the cellular lipid content decreased but the carbohydrate and protein contents increased. Under carbon limitation, the cellular lipid, protein, and carbohydrate contents remained relatively constant at the different dilution rates. Moreover, the cellular elemental composition was essentially identical under nitrogen and carbon limitation at a high dilution rate of 0.20h-1. We also analyzed the consumed carbon to nitrogen (C/N) under different nutrition conditions. The results indicated that the consumed C/N had a major influence on cell metabolism and product formation, which contributed to our understanding of the physiological characteristics of R. toruloides.

Molecular dynamics of FMRP and other RNA-binding proteins in MEG-01 differentiation: the role of mRNP complexes in non-neuronal development.

Asymmetrically differentiating cells are formed with the aid of RNA-binding proteins (RBPs), which can bind, stabilize, regulate, and transport target mRNAs. The loss of RBPs in neurons may lead to severe neurodevelopmental diseases such as the Fragile X Syndrome with the absence of the Fragile X Mental Retardation Protein (FMRP). Because the latter is ubiquitous and shares many similarities with other RBPs involved in the development of peripheral cells, we suggest that FMRP would have a role in the differentiation of all tissues where it is expressed. A MEG-01 differentiation model was, therefore, established to study the global developmental functions of FMRP. PMA induction of MEG-01 cells causes important morphological changes driven by cytoskeletal dynamics. Cytoskeleton change and colocalization analyses were performed by confocal microscopy and sucrose gradient fractionation. Total cellular protein content and de novo synthesis were also analyzed. Microtubular transport mediates the displacement of FMRP and other RBP-containing mRNP complexes towards regions of the cell in development. De novo protein synthesis decreases significantly upon differentiation and total protein content composition is altered. Because those results are comparable with those obtained in neurons, the absence of FMRP would have significant consequences in cells everywhere in the body. The latter should be further investigated to give a better understanding of the systemic implications of imbalances of FMRP and other functionally similar RBPs.

DUNALIELLA ЗАМГИЙН ФИЗИОЛОГИ, БИОХИМИЙН СУДАЛГААНЫ ДҮН

The genus Dunaliella is widely studied microalgae for its tolerance to extreme conditions, physiological aspects and many biotechnological applications, such as beta-carotene, protein, lipids and many other bioactive compounds. Physiological and biochemical studies are essential to fully explore the properties and possibilities of new isolates of Dunaliella.The aim of this study was to describe cellular morphology, growth rate and protein content of three Dunaliella strains, isolated from salty lakes in Mongolia. The cellular morphology, growth rates, protein contents were studied using microscopic analyses, Neubauer’s chamber, and micro kjeldahl method.Results showed that growths of all three Dunaliella cultures were progressed through lag phase at 3rd day. The growth of Dunaliella D-1 reached its peak on day 6, while Dunaliella D-6 and D-7 reached their stationary phase on day 7. Furthermore, the protein contents of dry biomass in Dunaliella D-1, D-6 and D-7 cultures were 62.2%, 36.1% and 38.15%, respectively. The highest protein content was found in Dunaliella D-1 culture, hence this culture could be used as protein rich supplement in further study.

Immunosuppressive and cytotoxic potential of aqueous leaves extract of Thlaspi arvense

Medicinal plants are traditionally used in Asian countries for treatment of various intracellular as well as extracellular infections. The present study was undertaken to investigate Thlaspi arvense for determining its immunosuppressive potential and cytotoxic activity in infected lysed human whole blood samples. For these studies, virally infected human whole blood samples (n = 5; symptoms such as sneezing, cough etc.) were collected and analysing its cytotoxicity of variable doses of aqueous leaves extract of Thlaspi arvense in the form of proliferation using Concanavalin (Con A; 2.5 µg/ml). In addition, immunosuppressive activity was also determined using lysed human whole blood samples were incubated with variable doses of aqueous extract in presence or absence of incomplete freunds adjuvant (IFA) pertaining to determine the total cellular (protein) content. In contrast, in vitro studies of aqueous extract of Thlaspi arvense were also determined using lactoferrin (100 µg/well) and ovalbumin (OVA, 100 µg/well) as coating antigen pertaining to antibody production using Elisa. The results showed that aqueous leaves extract of Thlaspi arvense showed markedly decline in Con A proliferation at higher doses. In addition, this aqueous extract also showed decline in total cellular content in presence or absence of IFA but it showed enhancement in antibody production in vitro against lactoferrin and OVA at higher doses. Overall the data showed that aqueous leaves extract of Thlaspi arvense displayed immunosuppressive and cytotoxic effect in virally infected human whole blood samples.

Anti-cancer activity of quercetin, gallic acid, and ellagic acid against hepg2 and hct 116 cell lines: in vitro

Hepatocarcinoma (HCC) and colorectal cancer (CRC) are the most malignant tumors with increasing incidences of morbidity and mortality globally.The aim of the present study was to determine in vitro antitumor activity of quercetin, gallic acid, and ellagic acid against HCC and CRC. This was achieved through testing the effects of these compounds on HepG2 and HCT 116 cells, ranging from cell growth inhibition assay and fold dose advantage, passing by mitochondrial activity assessment, to cellular protein content measurement. The efficacies of these compounds were compared with doxorubicin in case of HepG2 cells and 5-fluorouracil in case of HCT 116 cells. The present findings indicated that in both cell types, gallic acid elicited the most pronounced anti-cancer activity and mitochondrial dysfunction compared to quercetin and ellagic acid. Extensive research is warranted to assess the precise molecular mechanisms of these natural products in bridling HCC and CRC.

Immunological Exploration of Primary Metabolite Extracted From Aqueous Stem Extract of Caralluma Fimbriata

Objective: The objective of our study is to determined its immunological property of primary (protein) metabolite extracted from aqueous stem extract of Caralluma fimbriata against specific protein antigens (lactoferrin and Ovalbumin, OVA) pertaining to determine B cell production in vitro by ELISA (enzyme linked immunosorbent assay) and also measured T cell production in vitro using immature bone marrow cells of mice exposed with Incomplete freunds adjuvant (IFA) and Concanavalin (Con) A. Methods: For these studies, estimation of protein from stem extract of Caralluma fimbriata were determined and also determining the total cellular (protein) content in immature bone marrow cells along with variable concentration of Caralluma fimbriata in presence or absence of IFA. In addition, lactoferrin (100 µg/well) and weak antigen i.e. ovalbumin (OVA; 100 µg/well) pertaining to antibody (IgG) production were determined in vitro through Elisa and also measured its proliferative response using Con A (2.5 µg/ml). Results: The results showed that aqueous stem extract of Caralluma fimbriata showed the presence of protein (0.698 mg/ml; 10 µl) content and also showed the enhancement in total cellular (protein) content in presence or absence of IFA which is determined through NanoDrop method. In addition, this aqueous stem extract showed enhancement in anti-lactoferrin and anti-OVA IgG titre in vitro at higher doses as compared to control but there is sudden decline in bone marrow cell proliferation containing Con A at higher doses. Conclusion: Our data suggest that aqueous stem extract of Caralluma fimbriata may help to raise antibodies in vitro against lactoferrin and OVA but sudden decline in Con A proliferative response in bone marrow cells. In other words, aqueous stem extract containing primary metabolite of Caralluma fimbriata could be a potent immune enhancer of B cells in vitro but inhibitor of T cells at higher doses.

The reduction of IL-6 gene expression, pAKT, pERK1/2, pSTAT3 signaling pathways and invasion activity by gallic acid in prostate cancer PC3 cells.

Prostate cancer (PC) is one of the most common cancers among men. Progression of prostate cancer is associated with an increase in cellular level of interleukin-6 (IL-6). Gallic acid (GA) is a polyhydroxy phenolic compound which can inhibit the growth of cancer cells. The aim of this study was to evaluate the effects of GA treatment on cell viability, proliferation, invasion, IL-6 gene expression, IL-6 secretion, cellular levels of pSTAT3, pERK1/2, and pAKT signaling proteins in human prostate cancer PC3 cells. PC3 cells viability after treatment with GA (0-120μM) was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The expression of IL-6 was investigated using real-time polymerase chain reaction. Cellular concentration of pSTAT3, pERK1/2, and pAKT signaling proteins were determined by Western blotting technic. PC3 cells invasion was assessed by invasion assay test. Treatment with GA caused a significant decrease in cell viability, proliferation, invasion, cellular levels of pSTAT3, pERK1/2, and pAKT signaling proteins after 48h in a dose-dependent manner. The level of IL-6 and its gene expression decreased significantly in PC3 cells treated with GA. Our results show that IL-6 down-regulation and decreased IL-6 protein level in PC3 cells by GA resulted in diminishing of pSTAT3, pERK1/2, and pAKT signaling proteins which lead to the reduction of the cell survival, proliferation, and invasion in PC3 cells. Therefore, it seems that GA can be considered an anticancer agent in the treatment of prostate cancer.