Search results for: in vitro activation

Authors: Olaniyi Fawole, Umezuruike Opara

Abstract:

Co-products obtained from pomegranate juice processing contain high levels of polyphenols with potential high added values. From value-addition viewpoint, the aim of this study was to evaluate the stability of polyphenolic concentrations in pomegranate fruit co-products in different solvent extracts and assess the effect on the total antioxidant capacity using the FRAP, DPPH˙ and ABTS˙+ assays during simulated in vitro digestion. Pomegranate juice, marc and peel were extracted in water, 50% ethanol (50%EtOH) and absolute ethanol (100%EtOH) and analysed for total phenolic concentration (TPC), total flavonoids concentration (TFC) and total antioxidant capacity in DPPH˙, ABST˙+ and FRAP assays before and after in vitro digestion. Total phenolic concentration (TPC) and total flavonoid concentration (TFC) were in the order of peel > marc > juice throughout the in vitro digestion irrespective of the extraction solvents used. However, 50% ethanol extracted 1.1 to 12-fold more polyphenols than water and ethanol solvents depending on co-products. TPC and TFC increased significantly in gastric digests. In contrast, after the duodenal, polyphenolic concentrations decreased significantly (p < 0.05) compared to those obtained in gastric digests. Undigested samples and gastric digests showed strong and positive relationships between polyphenols and the antioxidant activities measured in DPPH, ABTS and FRAP assays, with correlation coefficients (r2) ranging between 0.930 – 0.990 whereas, the correlation between polyphenols (TPC and TFC) and radical cation scavenging activity (in ABTS) were moderately positive in duodenal digests. Findings from this study also showed that the concentration of pomegranate polyphenols and antioxidant thereof during in vitro gastro-intestinal digestion may not reflect the pre-digested phenolic concentration. Thus, this study highlights the need to provide biologically relevant information on antioxidants by providing data reflecting their stability and activity after in vitro digestion.

Keywords: by-product, DPPH, polyphenols, value addition

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Authors: Taraneh Estedlal

Abstract:

The objective of this study was to review in-vivo and in-vitro studies to compare the effectiveness of bulk-fill and conventional resin composites with regard to marginal adaptation, polymerization shrinkage, and other mechanical properties.PubMed and Scopus databases was investigated for in-vitro studies and randomized clinical trials comparing incidence of fractures, color stability, marginal adaptation, pain and discomfort, recurrent caries, occlusion, pulpal reaction, and proper proximal contacts of restorations made with conventional and bulk resins. The failure rate of conventional and flowable bulk-fill resin composites was not significantly different to sculptable bulk-fill resin composites. The objective of this study was to review in-vivo and in-vitro studies to compare the effectiveness of bulk-fill and conventional resin composites with regard to marginal adaptation, polymerization shrinkage, and other mechanical properties. PubMed and Scopus databases was investigated for in-vitro studies and randomized clinical trials comparing one of the pearlier mentioned properties between bulk-fill and control composites. Despite differences in physical and in-vitro properties, failure rate of conventional and flowable bulk-fill resin composites was not significantly different to sculptable bulk-fill resin composites.

Keywords: polymerization shrinkage, color stability, marginal adaptation, recurrent caries, occlusion, pulpal reaction

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Authors: Emily Schlebes, Christian Hundhausen, Jens W. Fischer

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The glycosaminoglycan hyaluronan (HA) is a major component of the extracellular matrix, typically produced by fibroblasts of the connective tissue but also by immune cells. Here, we investigated the capacity of human peripheral blood CD8 T cells from healthy donors to produce HA and to express HA receptors as well as HA degrading enzymes. Further, we evaluated the effect of pharmacological HA inhibition on CD8 T cell function. Using immunocytochemistry together with quantitative PCR analysis, we found that HA synthesis is rapidly induced upon antibody-induced T cell receptor (TCR) activation and almost exclusively mediated by HA synthase 3 (HAS3). TCR activation also resulted in the upregulation of HA receptors CD44, hyaluronan-mediated motility receptor (HMMR), and layilin (LAYN), although kinetics and strength of expression varied greatly between subjects. The HA-degrading enzymes HYAL1 and HYAL2 were detected at low levels and induced by cell activation in some individuals. Interestingly, expression of HAS3, HA receptors, and hyaluronidases were modulated by the proinflammatory cytokines IL-6 and IL-1bβ in most subjects. To assess the functional role of HA in CD8 T cells, we performed carboxyfluorescein succinimidyl ester (CFSE) based proliferation assays and cytokine analysis in the presence of the HA inhibitor 4- Methylumbelliferone (4-MU). Despite significant inter-individual variation with regard to the effective dose, 4-MU resulted in the inhibition of CD8 T cell proliferation and reduced release of TNF-α and IFN-γ. Collectively, these data demonstrate that human CD8 T cells respond to TCR stimulation with a synthesis of HA and expression of HA-related genes. They further suggest that HA inhibition may be helpful in interfering with pathogenic T cell activation in human disease.

Keywords: CD8 T cells, extracellular matrix, hyaluronan, hyaluronan synthase 3

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Authors: Gajera Lalit, Shah Pranav, Shah Shailesh

Abstract:

Venlafaxine hydrochloride has a short elimination half-life of 5 ± 2 hr, and absorption window in the upper part of gastrointestinal tract. The conventional tablets need to be administered two to three times a day and possess an oral bioavailability of 45%. The purpose of this study was to formulate gastroretentive effervescent floating tablets of Venlafaxine HCl. Different grades of HPMC namely K15M, K4M, K100M and E15LV were employed as swelling polymers whereas sodium bicarbonate was employed as gas generating agent. The direct compression method was employed for the formulation of tablets. The tablets were evaluated in terms of hardness, friability, weight variation, drug content, water uptake, in-vitro floating behavior and in-vitro drug release study. All the formulations exhibited very short floating lag time of < 1 min and total floating time of 12 hr. Formulation L3 containing 25 mg and 75 mg of HPMC E15 LV and HPMC K15M respectively exhibited complete drug release within 12 hrs.

Keywords: venlafaxine HCl, hydroxyl propyl methylcellulose, floating gastro retentive tablets, in-vitro drug release, non-fickian diffusion

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Authors: Zelikha Labbani

Abstract:

Since its creation in 1964 by Guha and Maheshwari in India on Datura innoxia Mill, in vitro androgenesis has become the method of choice in the production of doubled haploid in many species. However in durum wheat, the Doubled haploid plant breeding programs remained limited due to the low production of androgenetic embryos and converting them into fertile green plants. We describe here an efficient method for inducing embryos and regenerating green plants directly from isolated microspores of durum wheat.

Keywords: Durum wheat, haploid embryos, on in vitro, pretreatment

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Authors: S. Mohajer , R. M. Taha, M. Adel

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Developing our knowledge of when pineapple roots grow can lead to improved water, fertilizer applications, and more precise culture management. This paper presents current understanding of morphological traits in pineapple roots, highlighting studies using incubation periods and various solid MS media treated with different sucrose concentrations and pH, which directly assess in vitro environmental factors. Rooting parameters had different optimal sucrose concentrations and incubation periods. All shoots failed to root in medium supplemented with sucrose at 5 g/L and no roots formed within the first 45 days in medium enriched with sucrose at 10 g/L. After 75 days, all shoots rooted in medium enriched with 10 and 20 g/L sucrose. Moreover, MS medium supplied with 20 g/L sucrose resulted in the longest and the highest number of roots with 27.3 mm and 4.7, respectively. Root function, such as capacity for P and N uptake, declined rapidly with root length. As a result, the longer the incubation period, the better the rooting responses would be.

Keywords: environmental factors, in vitro rooting, pineapple, tissue culture

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Authors: Farah El Mohtadi, Richard d'Arcy, Nicola Tirelli

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Since 40 years, poly (ethylene glycol) (PEG) has been the gold standard in biomaterials and drug delivery, because of its combination of chemical and biological inertness. However, the possibility of its breakdown under oxidative conditions and the demonstrated development of anti-PEG antibodies highlight the necessity to develop carriers based on materials with increased stability in a challenging biological environment. Here, we describe the synthesis of polysulfide via anionic ring-opening polymerization. In vitro, the synthesized polymer was characterized by low toxicity and a level of complement activation (in human plasma) and macrophage uptake slightly lower than PEG and poly (2‐methyl-2‐oxazoline) (PMOX), of a similar size. Importantly, and differently from PEG, on activated macrophages, the synthesized polymer showed a strong and dose-dependent ROS scavenging activity, which resulted in the corresponding reduction of cytokine production. Therefore, the results from these studies show that polysulfide is highly biocompatible and are potential candidates to be used as an alternative to PEG for various applications in nanomedicine.

Keywords: PEG, low toxicity, ROS scavenging, biocompatible

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Authors: Yamani Amal, Lazaae Jamila, Elachouri Mostafa

Abstract:

Plants and their active agents – especially polyphenols – may have a principal role in the treatment of diseases that result from the defect of physiological antioxidant mechanisms. Bryonia dioica is well known in Moroccan traditional medicine for alleviatin pain and traiting many diseases. We have focused on plant belonging to Cucurbitaceae Family from around the world to understand their therapeutic uses and their potential antioxidant activities Although several biological activities and Chemical composition of Bryonia dioica are well characterized, no direct, in vitro study, of this natural product examined the antioxydant effect of the extract from the roots of Bryonia dioica. The aim of this study was to determine in vitro antioxidant activity of the B.dioica root, using antioxidant analysis methods based on determination of Hydroxyradical Scavenging, 1,1-diphenyl-2-picrylhydrazine (DPPH) radical scavenging, Hydrogenperoxide Scavenging and Nitric Oxide Scavenging. In this study, it was demonstrated, that, B. dioica root extract showed excellent antioxidant properties. This investigation showed that the roots of this plant contain potent natural scavengers R. It may represent an interesting source of antioxidant phenolics that may favour the extension of their cultivation as new source of natural antioxidants in addition to containing high quality proteins for human or animal nutrition. Therefore, there is need for all stakeholders on the Morocco to strive towards taking advantage of our enormous biodiversity resources to free our people from diseases, abject poverty and stagnation.

Keywords: Morocco, bryoniadioica, in vitro, antioxydant

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Authors: Hanjun Zhao, Ke Zhang, Bojian Zheng

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Background: So far, there is no universal antiviral agent which can inhibit multiple viral infections. More and more drug-resistant viral strains emerge after the antiviral drug application for treatment. Defensins are the front line of host innate immunity and have broad spectrum antibacterial and antiviral effects. However, there is limited data to show if these defensins have good antiviral activity in vivo and what the antiviral mechanism is. Subjects: To investigate a peptide with widespread antivirus activity in vitro and in vivo and illustrate the antiviral mechanism. Methods: Antiviral peptide library designed from mouse beta defensins was synthesized by the company. Recombinant beta defensin was obtained from E. coli. Antiviral activity in vitro was assayed by plaque assay, qPCR. Antiviral activity in vivo was detected by animal challenge with 2009 pandemic H1N1 influenza A virus. The antiviral mechanism was assayed by western blot, ELISA, and qPCR. Conclusions: We identify a new peptide which has widespread effects against multiple viruses (H1N1, H5N1, H7N9, MERS-CoV) in vitro and has efficient antivirus activity in vivo. This peptide inhibits viral entry into target cells and subsequently blocks viral replication. The in vivo study of the antiviral peptide against other viral infections and the investigation of its more detail antiviral mechanism are ongoing.

Keywords: antiviral peptide, defensin, Influenza A virus, mechanism

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Authors: J. Linjikao, P. Inthima, A. Kongbangkerd

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In vitro conservation of orchid germplasm provides an effective technique for ex situ conservation of orchid diversity. In this study, an efficient protocol for in vitro conservation of Vandopsis lissochiloides (Gaudich.) Pfitz. plantlet under slow growth conditions was investigated. Plantlets were cultured on different strength of Vacin and Went medium (½VW and ¼VW) supplemented with different concentrations of mannitol (0, 2, 4, 6 and 8%), sucrose (0 and 3%) and 50 g/L potato extract, 150 mL/L coconut water. The cultures were incubated at 25±2 °C and maintained under 20 µmol/m²s light intensity for 24 weeks without subculture. At the end of preservation period, the plantlets were subcultured to fresh medium for growth recovery. The results found that the highest leaf number per plantlet could be observed on ¼VW medium without adding sucrose and mannitol while the highest root number per plantlet was found on ½VW added with 3% sucrose without adding mannitol after 24 weeks of in vitro storage. The results showed that the maximum number of leaves (5.8 leaves) and roots (5.0 roots) of preserved plantlets were produced on ¼VW medium without adding sucrose and mannitol. Therefore, ¼VW medium without adding sucrose and mannitol was the best minimum growth conditions for medium-term storage of V. lissochiloides plantlets.

Keywords: preservation, vandopsis, germplasm, in vitro

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Authors: Marcel Verwey, Anna M. Joubert, Elsie M. Nolte, Wolfgang Dohle, Barry V. L. Potter, Anne E. Theron

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A tetrahydroisoquinolinone (THIQ) core can be used to mimic the A,B-ring of colchicine site-binding microtubule disruptors such as 2-methoxyestradiol in the design of anti-cancer agents. Steroidomimeric microtubule disruptors were synthesized by introducing C'2 and C'3 of the steroidal A-ring to C'6 and C'7 of the THIQ core and by introducing a decorated hydrogen bond acceptor motif projecting from the steroidal D-ring to N'2. For this in vitro study, four non-steroidal THIQ-based analogues were investigated and comparative studies were done between the non-sulphamoylated compound STX 3450 and the sulphamoylated compounds STX 2895, STX 3329 and STX 3451. The objective of this study was to investigate the modes of cell death induced by these four THIQ-based analogues in A549 lung carcinoma epithelial cells and metastatic breast adenocarcinoma MDA-MB-231 cells. Cytotoxicity studies to determine the half maximal growth inhibitory concentrations were done using spectrophotometric quantification via crystal violet staining and lactate dehydrogenase (LDH) assays. Microtubule integrity and morphologic changes of exposed cells were investigated using polarization-optical transmitted light differential interference contrast microscopy, transmission electron microscopy and confocal microscopy. Flow cytometric quantification was used to determine apoptosis induction and the effect that THIQ-based analogues have on cell cycle progression. Signal transduction pathways were elucidated by quantification of the mitochondrial membrane integrity, cytochrome c release and caspase 3, -6 and -8 activation. Induction of autophagic cell death by the THIQ-based analogues was investigated by morphological assessment of fluorescent monodansylcadaverine (MDC) staining of acidic vacuoles and by quantifying aggresome formation via flow cytometry. Results revealed that these non-steroidal microtubule disrupting analogues inhibited 50% of cell growth at nanomolar concentrations. Immunofluorescence microscopy indicated microtubule depolarization and the resultant mitotic arrest was further confirmed through cell cycle analysis. Apoptosis induction via the intrinsic pathway was observed due to depolarization of the mitochondrial membrane, induction of cytochrome c release as well as, caspase 3 activation. Potential involvement of programmed cell death type II was observed due to the presence of acidic vacuoles and aggresome formation. Necrotic cell death did not contribute significantly, indicated by stable LDH levels. This in vitro study revealed the induction of the intrinsic apoptotic pathway as well as possible involvement of autophagy after exposure to these THIQ-based analogues in both MDA-MB-231- and A549 cells. Further investigation of this series of anticancer drugs still needs to be conducted to elucidate the temporal, mechanistic and functional crosstalk mechanisms between the two observed programmed cell deaths pathways.

Keywords: apoptosis, autophagy, cancer, microtubule disruptor

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Authors: Rohita Gupta, G. S. Brah, R. Verma, C. S. Mukhopadhayay

Abstract:

Although chicken strains show differences in susceptibility to a number of diseases, the underlying immunological basis is yet to be elucidated. In the present study, heterophils were subjected to LPS stimulation and total RNA extraction, further differential gene expression was studied in broiler, layer and indigenous Aseel strain by Real Time RT-PCR at different time periods before and after induction. The expression of the 14 AvBDs and chTLR 1, 2, 3, 4, 5, 7, 15 and 21 was detectable in heterophils. The expression level of most of the AvBDs significantly increased (P<0.05) 3 hours post in vitro lipopolysaccharide challenge. Higher expression level and stronger activation of most AvBDs, NFkB-1 and IRF-3 in heterophils was observed with the stimulation of LPS in layer compared to broiler, and in Aseel compared to both layer and broiler. This investigation will allow more refined interpretation of immuno-genetic basis of the variable disease resistance/susceptibility in divergent stock of chicken including indigenous breed. Moreover, this study will be helpful in formulation of strategy for isolation of antimicrobial peptides from heterophils.

Keywords: differential expression, heterophils, cytokines, defensin, TLR

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Authors: Pinar Ercan, Sedef N. El

Abstract:

The goals of this study were to determine the total anthocyanin and procyanidin contents and their in vitro bioaccessibilities of apple, red grape and cinnamon by a static in vitro digestion method reported by the COST FA1005 Action INFOGEST, as well as in vitro inhibitory effects of these food samples on starch and lipid digestive enzymes. While the highest total anthocyanin content was found in red grape (164.76 ± 2.51 mg/100 g), the highest procyanidin content was found in cinnamon (6432.54±177.31 mg/100 g) among the selected food samples (p<0.05). The anthocyanin bioaccessibilities were found as 10.23±1 %, 8.23±0.64 %, and 8.73±0.70 % in apple, red grape, and cinnamon, respectively. The procyanidin bioaccessibilities of apple, red grape, and cinnamon were found as 17.57±0.71 %, 14.08±0.74 % and 18.75±1.49 %, respectively. The analyzed apple, red grape and cinnamon showed the inhibitory activity against α-glucosidase (IC50 544.27±21.94, 445.63±15.67, 1592±17.58 μg/mL, respectively), α-amylase (IC50 38.41±7.26, 56.12±3.60, 3.54±0.86 μg/mL, respectively), and lipase (IC50 52.65±2.05, 581.70±54.14, 49.63±2.72 μg/mL, respectively). Red grape sample showed the highest inhibitory activity against α-glucosidase, cinnamon showed the highest inhibitory activity against α-amylase and lipase according to IC50 (concentration of inhibitor required to produce a 50% inhibition of the initial rate of reaction) and Catechin equivalent inhibition capacity (CEIC50) values. This study reported that apple, grape and cinnamon samples can inhibit the activity of digestive enzymes in vitro. The consumption of these samples would be used in conjunction with a low-calorie diet for body weight management.

Keywords: anthocyanin, α-amylase, α-glucosidase, lipase, procyanidin

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Authors: Alhulw H. Alshammari, Ahmed Iraqi, S. A. Saad, T. A. Taha

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In this work, we present for the first time the study of Raman spectra and dielectric relaxation of polyester polymer-CoFe₂O₄ (5.0, 10.0, 15.0, and 20.0 wt%) nanocomposites. Raman spectroscopy was applied as a sensitive structural identification technique to characterize the polyester-CoFe₂O₄ nanocomposites. The images of AFM confirmed the uniform distribution of CoFe₂O₄ inside the polymer matrix. Dielectric relaxation was employed as an important analytical technique to obtain information about the ability of the polymer nanocomposites to store and filter electrical signals. The dielectric relaxation analyses were carried out on the polyester-CoFe₂O₄ nanocomposites at different temperatures. An increase in dielectric constant ε₁ was observed for all samples with increasing temperatures due to the alignment of the electric dipoles with the applied electric field. In contrast, ε₁ decreased with increasing frequency. This is attributed to the difficulty for the electric dipoles to follow the electric field. The α relaxation peak that appeared at a high frequency shifted to higher frequencies when increasing the temperature. The activation energies for Maxwell-Wagner Sillar (MWS) changed from 0.84 to 1.01 eV, while the activation energies for α relaxations were 0.54 – 0.94 eV. The conduction mechanism for the polyester- CoFe₂O₄ nanocomposites followed the correlated barrier hopping (CBH) model.

Keywords: AC conductivity, activation energy, dielectric permittivity, polyester nanocomposites

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Authors: Philip Vercoe, Ali Hardan

Abstract:

Samples of 29 Australian wild natives and exotic plants were tested in vitro batch rumen culture system for their methanogenic characteristics and potential usage as feed or antimicrobial to enhance sustainable livestock ruminant production system. The plants were tested for their in vitro rumen fermentation end products properties which include: methane production, total gas pressure, concentrations of total volatile fatty acids, ammonia, and acetate to propionate ratio. All of the plants were produced less methane than the positive control (i.e., oaten chaff) in vitro. Nearly 50 % of plants inhibiting methane by over 50% in comparison to the control. Eremophila granitica had the strongest inhibitory effect about 92 % on methane production comparing with oaten chaff. The exotic weed Arctotheca calendula (Capeweed) had the highest concentration of volatile fatty acids production as well as the highest in total gas pressure among all plants and the control. Some of the acacia species have the lowest production of total gas pressure. The majority of the plants produced more ammonia than the oaten chaff control. The plant species that produced the most ammonia was Codonocarpus cotinifolius, producing over 3 times as much methane as oaten chaff control while the lowest was Eremophila galeata. There was strong positive correlation between methane production and total gas production as well as between total gas production and the concentration of VFA produced with R² = 0.74, R² = 0.84, respectively. While there was weak positive correlation between methane production and the acetate to propionate ratio as well as between the concentration of VFA produced and methane production with R² = 0.41, R² = 0.52, respectively.

Keywords: in vitro Rumen Fermentation, methane, wild Australian native plants, forages

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Authors: Yaowaporn Sangsen, Kittisak Likhitwitayawuid, Boonchoo Sritularak, Kamonthip Wiwattanawongsa, Ruedeekorn Wiwattanapatapee

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Novel solid lipid nanoparticles (SLNs) were developed to improve oral bioavailability of oxyresveratrol (OXY). The SLNs were prepared by a high speed homogenization technique, at an effective speed and time, using Compritol® 888 ATO (5% w/w) as the solid lipid. The appropriate weight proportions (0.3% w/w) of OXY affected the physicochemical properties of blank SLNs. The effects of surfactant types on the properties of the formulations such as particle size and entrapment efficacy were also investigated. Conclusively, Tween 80 combined with soy lecithin was the most appropriate surfactant to stabilize OXY-loaded SLNs. The mean particle size of the optimized formulation was 134.40 ± 0.57 nm. In vitro drug release study, the selected S2 formulation showed a retarded release profile for OXY with no initial burst release compared to OXY suspension in the simulated gastrointestinal fluids. Therefore, these SLNs could provide a suitable system to develop for the oral OXY delivery.

Keywords: solid lipid nanoparticles, physicochemical properties, in vitro drug release, oxyresveratrol

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Authors: Nagarjuna Remalli, Robert Brandt

Abstract:

Down-sizing of combustion engines in automobiles are prevailed owing to required increase in efficiency. This leads to a stress increment on valve springs, which affects their intended function due to an increase in relaxation. High strength martensitic steels are used for valve spring applications. Recent investigations unveiled that low temperature creep (LTC) in martensitic steels obey a logarithmic creep law. The exhaustion theory links the logarithmic creep behavior to an activation energy which is characteristic for any given time during creep. This activation energy increases with creep strain due to barriers of low activation energies exhausted during creep. The assumption of the exhaustion theory is that the material is inhomogeneous in microscopic scale. According to these assumptions it is anticipated that small obstacles (e. g. ε–carbides) having a wide range of size distribution are non-uniformly distributed in the materials. X-ray diffraction studies revealed the presence of ε–carbides in high strength martensitic steels. In this study, high strength martensitic steels that are crept in the temperature range of 75 – 150 °C were investigated with the aid of a transmission electron microscope for the evidence of an inhomogeneous distribution of obstacles having different size to examine the validation of exhaustion theory.

Keywords: creep mechanisms, exhaustion theory, low temperature creep, martensitic steels

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Authors: Olusegun O. Ikusika, Conference T. Mpendulo

Abstract:

The abundance of fossil shell flour (FSF) globally has increased interest in its use as a natural feed additive in livestock diets. Therefore, identifying its optimum inclusion levels in livestock production is essential for animal productivity. This study investigated the effects of various fossil shell flour (FSF) inclusion levels on in vitro digestibility, relative feed values, and rumen parameters of Dohne-Merino wethers. Twenty-four fistulated wethers with an average body weight of 20 ± 1•5 kg in a complete randomized design of four treatments having six wethers per treatment were used. They were fed a basal diet without fossil shell flour (control, 0%) or with the addition of 2% FSF (T2), 4% FSF(T3), and 6% FSF (T4) of diet DM for 35 days, excluding 14 days adaptation period. The results showed that increasing FSF levels had no effect on ruminal T0C or pH, but Ammonia-N increased (P<0.01) with increasing FSF. The total molar concentrations of volatile fatty acids (VFA) decreased (P<0.05) with increasing levels of FSF. Acetic: propionic ratio decreased except at the 4 % inclusion level. IVTDDM, IVTDNDF and IVTDADF decreased up till 4% FSF inclusion but tended to increase (P = 0.06) at 6% inclusion. Relative feed values of the diets tended to increase (P=0.07) by adding fossil shell flour. In conclusion, adding FSF to the diets of Dohne-Merino wether up to 6% FSF inclusion rates did not improve IVTDDM (In vitro true digestibility dry matter), IVTDNDF (In vitro true digestibility neutral detergent fiber), and IVTDADF (In vitro true digestibility acid detergent fiber). However, a small increment of rumen nitrogen with no adverse effects on the rumen parameters was observed. The relative feed value (RFV) moved the feed from good to premium when supplemented. Therefore, FSF supplementation could improve feed value and maintain a normal range of rumen parameters for the effective functionality of the rumen.

Keywords: fossil shell flour, rumen parameters, in vitro digestibility, feed quality, dohne-merino sheep

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Authors: Seyedahmad Hosseini, Mohammadjavad Sotoudeheian

Abstract:

Introduction: Allergic asthma is a heterogeneous immuno-inflammatory disease based on Th-2-mediated inflammation. Histopathologic abnormalities of the airways characteristic of asthma include epithelial damage and subepithelial collagen deposition. Objectives: Human bronchial epithelial cell genome expression of TNF‑α, IL‑6, ICAM‑1, VCAM‑1, nuclear factor (NF)‑κB signaling pathways up-regulate during inflammatory cascades. Moreover, immunofluorescence assays confirmed the nuclear translocation of NF‑κB p65 during inflammatory responses. An absolute LDH leakage assays suggestedLPS-inducedcells injury, and the associated mechanisms are co-incident events. LPS-induced phosphorylation of ERKand JNK causes inflammation in epithelial cells through inhibition of ERK and JNK activation and NF-κB signaling pathway. Furthermore, the inhibition of NF-κB mRNA expression and the nuclear translocation of NF-κB lead to anti-inflammatory events. Likewise, activation of SUMF2 which inhibits IL-13 and reduces Th2-cytokines, NF-κB, and IgE levels to ameliorate asthma. On the other hand, TNFα-induced mucus production reduced NF-κB activation through inhibition of the activation status of Rac1 and IκBα phosphorylation. In addition, bradykinin B2 receptor (B2R), which mediates airway remodeling, regulates through NF-κB. Bronchial B2R expression is constitutively elevated in allergic asthma. In addition, certain NF-κB -dependent chemokines function to recruit eosinophils in the airway. Besides, bromodomain containing 4 (BRD4) plays a significant role in mediating innate immune response in human small airway epithelial cells as well as transglutaminase 2 (TG2), which is detectable in saliva. So, the guanine nucleotide-binding regulatory protein α-subunit, Gα16, expresses a κB-driven luciferase reporter. This response was accompanied by phosphorylation of IκBα. Furthermore, expression of Gα16 in saliva markedly enhanced TNF-α-induced κB reporter activity. Methods: The applied method to form NF-κB activation is the electromobility shift assay (EMSA). Also, B2R-BRD4-TG2 complex detection by immunoassay method within saliva with EMSA of NF-κB activation may be a novel biomarker for asthma diagnosis and follow up. Conclusion: This concept introduces NF-κB signaling pathway as potential asthma biomarkers and promising targets for the development of new therapeutic strategies against asthma.

Keywords: NF-κB, asthma, saliva, T-helper

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Authors: Rohita Gupta, G. S. Brah, R. Verma, C. S. Mukhopadhayay

Abstract:

Although chicken strains show differences in susceptibility to a number of diseases, the underlying immunological basis is yet to be elucidated. In the present study, heterophils were subjected to LPS stimulation and total RNA extraction, further differential gene expression was studied in broiler, layer and indigenous Aseel strain by Real Time RT-PCR at different time periods before and after induction. The expression of the 14 AvBDs and chTLR 1, 2, 3, 4, 5, 7, 15 and 21 was detectable in heterophils. The expression level of most of the AvBDs significantly increased (P<0.05) 3 hours post in vitro lipopolysaccharide challenge. Higher expression level and stronger activation of most AvBDs, NFkB-1 and IRF-3 in heterophils was observed, with the stimulation of LPS in layer compared to broiler, and in Aseel compared to both layer and broiler. This investigation will allow more refined interpretation of immuno-genetic basis of the variable disease resistance/susceptibility in divergent stock of chicken including indigenous breed. Moreover this study will be helpful in formulation of strategy for isolation of antimicrobial peptides from heterophils.

Keywords: differential expression, heterophils, cytokines, defensin, TLR

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Authors: M. Sepahvand, M. Khorushy

Abstract:

Gladiolus, being a cormous plant, it is principally propagated by the natural multiplication of new corms and cormels. In order to obtain callus from segmented corm which was obtained from in vitro culture, callus formation media were MS media supplemented with 4 levels of hormones such as 1.0 mg l-1 NAA + 0.5 mg l-1 BAP, 0.5 mg l-1 NAA + 0.25 mg l-1 BAP, 1.0 mg l-1 2, 4-D + 0.5 mg l-1 BAP, and 0.5 mg l-1 2, 4-D + 0.25 mg l-1 BAP. The results showed that the most weight of callus (2.28 g) was produced in MS callus formation media which were supplemented with 1.0 mg l-1 NAA + 0.5 mg l-1 BAP. This experiment was carried out in randomized completely design with 3 replications and each treatment with six jars. In second experiment for regeneration of callus, a factorial experiment in the form of randomized complete design with 12 treatments and 3 replications and each replication with six jars was carried out. The treatments consisted of callus culture media in 4 levels and regeneration culture media in 3 levels [control (no PGRs), MS with 0.2 mg l-1 BAP + 0.1 mg l-1 Kin + 0.01 mg l-1 NAA, and MS with 0.2 mg l-1 BAP + 0.05 mg l-1 Kin + 0.01 mg l-1 NAA]. The results showed that the best regeneration media were MS media which were supplemented with 0.2 mg l-1 BAP + 0.1 mg l-1 Kin. + 0.01 mg l-1 NAA that had the highest number of shoots (7/83 N), and shoot length (7/3 cm).

Keywords: regeneration, Segmented corm explant, callus, in vitro, gladiolus cv. white prosperity

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Authors: Venoos Iman, Suzita Mohd Noor, Syam Mohan, Mohamad Ibrahim Noordin

Abstract:

Girinimbine, a carbazole alkaloid was isolated from the stem bark and root of Murraya koenigii and its structure and purity was identified by HPLC and LC-MS. Here we report that Girinimbine strongly inhibit angiogenesis activity both in vitro and in vivo. MTT result showed that girinimbine inhibits cell proliferation of the HUVECS cell line in vitro. Result of endothelial cell invasion, migration, tube formation and wound healing assays also demonstrated significant time and does dependent inhibition by girinimbine. Moreover, girinibine mediates its anti-angiogenic activity through up- and down-regulation of angiogenic and anti-aniogenic proteins. Furthermore, anti-angiogenic potential of girinimbine was evidenced in vivo on zebrafish model. Girinimbine inhibited neo-vessels formation in zebrafish embryos during 24 hours exposure time. Together, these results demonstrated for the first time that girinimbine could effectively suppress angiogenesis and strongly suggest that it might be a novel angiogenesis inhibitor.

Keywords: anti-angiogenic, carbazole alkaloid, girinimbine, zebrafish

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Authors: Jay J. Vora, Vishvesh J. Badheka

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This work attempts to investigate the effect of oxide fluxes on 6mm thick Reduced Activation ferritic/martensitic steels (RAFM) during Activated TIG (A-TIG) welding. Six different fluxes Al₂O₃, Co₃O₄, CuO, HgO, MoO₃, and NiO were mixed with methanol for conversion into paste and bead-on-plate experiments were then carried out. This study, systematically investigates the influence of oxide-based flux powder and carrier solvent composition on the weld bead shape, geometric shape of weld bead and dominant depth enhancing mechanism in tungsten inert gas (TIG) welding of reduced activation ferritic/martensitic (RAFM) steel. It was inferred from the study that flux Co₃O₄ and MoO₃ imparted full and secure (more than 6mm) penetration with methanol owing to dual mechanism of reversed Marangoni and arc construction. The use of methanol imparted good spreadabilty and coverability and ultimately higher peak temperatures were observed with its use owing to stronger depth enhancing mechanisms than use of acetone with same oxide fluxes and welding conditions.

Keywords: A-TIG, flux, oxides, penetration, RAFM, temperature, welding

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Authors: Jianli Dong, Fangling Xu, Gengming Huang

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Human endogenous retroviral elements (HERVs) comprise approximately 8% of the human genome. They are thought to be germline-integrated genetic remnants of retroviral infections. Although HERV sequences are highly defective, some, especially the K type (HERV-K), have been shown to be expressed and may have biological activities in the pathogenesis of cancer, chronic inflammation and autoimmune diseases. We found that HERV-K GAG and ENV proteins were strongly expressed in pleomorphic melanoma cells. We also detected a critical role of HERV-K ENV in mediating intercellular fusion and colony formation of melanoma cells. Interestingly, we found that levels of HERV-K GAG and ENV expression correlated with the activation of ERK and loss of p16INK4A in melanoma cells, and inhibition of MEK or CDK4, especially in combination, reduced HERV-K expression in melanoma cells. We also performed a reverse transcription-polymerase chain reaction (RT-PCR) assay using DNase I digestion to remove “contaminating” HERV-K genomic DNA and examined HERV-K RNA expression in plasma samples from HIV-1 infected individuals. We found a covariation between HERV-K RNA expression and CD4 cell counts in HIV-1 positive samples. Although a causal link between HERV-K activation and melanoma development, and between HERV-K activation, HIV-1 infection and CD4 cell count have yet to be determined, existing data support the further research efforts in HERV-K.

Keywords: CD4 cell, HERV-K, HIV-1, melanoma

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Authors: Kefale W. Yizengaw, Delele Worku Ayele, Jyh-Chiang Jiang

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The catalytic co-conversion of CO₂ and CH₄ to value-added compounds has become one of the promising approaches to addressing global climate change by having valuable fossil fuels. Thedirect co-conversion of CO₂ and CH₄ to value-added compounds is attractive but tremendously challenging because of both molecules' thermodynamic stability and kinetic inertness. In the present study, a single iridium atom doped and a single oxygen atom defect hematite (110)surface model catalyst, which can comprehend direct C–O coupling based on simultaneous activation of CO2 and CH4 was studied using density functional theory plus U (DFT + U)calculations. The presence of dual active sites on the Ir/Fe₂O₃(110)-OV surface catalyst enablesCO₂ activation on the Ir site and CH₄ activation at the defect site. The electron analysis for the theco-adsorption of CO₂ and CH₄ deals with the electron redistribution on the surface and clearly shows the synergistic effect for simultaneous CO₂ and CH₄ activation on Ir/α- Fe₂O₃(110)-OVsurface. The microkinetic analysis shows that the dissociation of CH4 to CH3 * and H* plays an excellent role in the C–O coupling. The coverage analysis for the intermediate products of the microkinetic simulation results indicates that C–O coupling is the reaction limiting step. Finally, after the CH₃O* intermediate product species is produced, the radical hydrogen species spontaneously diffuse to the CH3O* intermediate product to form methanol at around 490 [K]. The present work provides mechanistic and kinetic insights into the direct C–O coupling of CO₂and CH₄, which could help design more-efficient catalysts.

Keywords: co-conversion, C–O coupling, doping, oxygen vacancy, microkinetic

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Authors: Virgil Miranda, Gissele Mosqueda, Pablo Delgado, Yimesker Yihun

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Muscular fatigue affects the muscle activation that is needed for producing the desired clinical outcome. Integrating optimal muscle relaxation periods into a variety of health care rehabilitation protocols is important to maximize the efficiency of the therapy. In this study, four muscle relaxation periods (30, 60, 90, and 120 seconds) and their effectiveness in producing consistent muscle activation of the muscle biceps brachii between sets of elbow flexion and extension task was investigated among a sample of 10 subjects with no disabilities. The same resting periods were then utilized in a controlled exoskeleton-based exercise for a sample size of 5 subjects and have shown similar results. On average, the muscle activity of the biceps brachii decreased by 0.3% when rested for 30 seconds, and it increased by 1.25%, 0.76%, and 0.82% when using muscle relaxation periods of 60, 90, and 120 seconds, respectively. The preliminary results suggest that a muscle relaxation period of about 60 seconds is needed for optimal continuous muscle activation within rehabilitation regimens. Robot-based rehabilitation is good to produce repetitive tasks with the right intensity, and knowing the optimal resting period will make the automation more effective.

Keywords: rest intervals, muscle biceps brachii, robot rehabilitation, muscle fatigue

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Authors: Ronald Croce, Timothy Quinn, John Miller

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Incomplete activation, or activation failure, of motor units during maximal voluntary contractions is often referred to as muscle inhibition (MI), and is defined as the inability of the central nervous system to maximally drive a muscle during a voluntary contraction. The purpose of the present study was to assess the interrelationship amongst peak torque (PT), muscle inhibition (MI; incomplete activation of motor units), and voluntary muscle activation (VMA) of the quadriceps’ muscle group as a function of knee angle and muscle length during maximal voluntary isometric contractions (MVICs). Nine young adult males (mean + standard deviation: age: 21.58 + 1.30 years; height: 180.07 + 4.99 cm; weight: 89.07 + 7.55 kg) performed MVICs in random order with the knee at 15, 55, and 95° flexion. MI was assessed using the interpolated twitch technique and was estimated by the amount of additional knee extensor PT evoked by the superimposed twitch during MVICs. Voluntary muscle activation was estimated by root mean square amplitude electromyography (EMGrms) and mechanomyography (MMGrms) of agonist (vastus medialis [VM], vastus lateralis [VL], and rectus femoris [RF]) and antagonist (biceps femoris ([BF]) muscles during MVICs. Data were analyzed using separate repeated measures analysis of variance. Results revealed a strong dependency of quadriceps’ PT (p < 0.001), MI (p < 0.001) and MA (p < 0.01) on knee joint position: PT was smallest at the most shortened muscle position (15°) and greatest at mid-position (55°); MI and MA were smallest at the most shortened muscle position (15°) and greatest at the most lengthened position (95°), with the RF showing the greatest change in MA. It is hypothesized that the ability to more fully activate the quadriceps at short compared to longer muscle lengths (96% contracted at 15°; 91% at 55°; 90% at 95°) might partly compensate for the unfavorable force-length mechanics at the more extended position and consequent declines in VMA (decreases in EMGrms and MMGrms muscle amplitude during MVICs) and force production (PT = 111-Nm at 15°, 217-NM at 55°, 199-Nm at 95°). Biceps femoris EMG and MMG data showed no statistical differences (p = 0.11 and 0.12, respectively) at joint angles tested, although there were greater values at the extended position. Increased BF muscle amplitude at this position could be a mechanism by which anterior shear and tibial rotation induced by high quadriceps’ activity are countered. Measuring and understanding the degree to which one sees MI and VMA in the QF muscle has particular clinical relevance because different knee-joint disorders, such ligament injuries or osteoarthritis, increase levels of MI observed and markedly reduced the capability of full VMA.

Keywords: electromyography, interpolated twitch technique, mechanomyography, muscle activation, muscle inhibition

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Authors: Essia Sebai, Amel Abidi, Hayet benyeddem, Akkari Hafidh

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Herbal extracts are of particular interest to the drug industry; essential oil with significant anthelmintic activity has the potential to be used as an alternative to conventional chemical drugs. In the present study, we describe the chemical profile of Laurus nobilis essential oil (EO), the in vitro anthelmintic activity of laurel oil against Haemonchus contortus and its in vivo anthelmintic effect against the murine helminth parasite model Heligmosomoides polygyrus. The chromatographic profile of L. nobilis (EO) extracted from the leaves of L. nobilis has shown the presence of monoterpenes 1,8-cineol (Eucalyptol) (29.47%), D-Limonène (18.51%) and Linalool (10.84%) in high fractions. The in vitro anthelmintic potential was expressed by an ovicidal effect against H. contortus egg hatching with an inhibition value of 3.23 mg/mL and 87.5% of immobility of adult worms after 8 hours of exposure to 8 mg/mL of L. nobilis EO. Regarding the in vivo anthelmintic potential, L. nobilis (EO) at 2400 mg/kg completely eliminated the egg output of H. polygyrus after seven days of oral treatment, together with a 79.2% of reduction in total worm counts. Based on the obtained funding, L. nobilis EO showed promising in vitro and in vivo anthelmintic capacities against gastrointestinal parasites.

Keywords: lauris nobilis, anthelmintic, haemonchus, pylogyrus

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Authors: Apoorva Vinod, Archana Mathur, Snehanshu Saha

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Though there exists a plethora of Activation Functions (AFs) used in single and multiple hidden layer Neural Networks (NN), their behavior always raised curiosity, whether used in combination or singly. The popular AFs –Sigmoid, ReLU, and Tanh–have performed prominently well for shallow and deep architectures. Most of the time, AFs are used singly in multi-layered NN, and, to the best of our knowledge, their performance is never studied and analyzed deeply when used in combination. In this manuscript, we experiment with multi-layered NN architecture (both on shallow and deep architectures; Convolutional NN and VGG16) and investigate how well the network responds to using two different AFs (Sigmoid-Tanh, Tanh-ReLU, ReLU-Sigmoid) used alternately against a traditional, single (Sigmoid-Sigmoid, Tanh-Tanh, ReLUReLU) combination. Our results show that using two different AFs, the network achieves better accuracy, substantially lower loss, and faster convergence on 4 computer vision (CV) and 15 Non-CV (NCV) datasets. When using different AFs, not only was the accuracy greater by 6-7%, but we also accomplished convergence twice as fast. We present a case study to investigate the probability of networks suffering vanishing and exploding gradients when using two different AFs. Additionally, we theoretically showed that a composition of two or more AFs satisfies Universal Approximation Theorem (UAT).

Keywords: activation function, universal approximation function, neural networks, convergence

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Authors: Leda Maria Costa Pereira, Maria Denise Lopes, Nucharin Songsasen

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Mammalian ovaries contain thousands of follicles that eventually degenerate or die after culture in vitro. Caspase-3 is a key enzyme that regulating cell death. Our objective was to examine the influence of anti-apoptotic drug Z-VAD-FMK (pan-caspase inhibitor) on in vitro viability of dog follicles within the ovarian cortex. Ovaries were obtained from prepubertal (age, 2.5–6 months) and adult (age, 8 months to 2 years) bitches and ovarian cortical fragments were recovered. The cortices were then incubated on 1.5% (w/v) agarose gel blocks within a 24-wells culture plate (three cortical pieces/well) containing Minimum Essential Medium Eagle - Alpha Modification (Alpha MEM) supplemented with 4.2 µg/ml insulin, 3.8 µg/ml transferrin, 5 ng/ml selenium, 2 mM L-glutamine, 100 µg/mL of penicillin G sodium, 100 µg/mL of streptomycin sulfate, 0.05 mM ascorbic acid, 10 ng/mL of FSH and 0.1% (w/v) polyvinyl alcohol in humidified atmosphere of 5% CO2 and 5% O2. The cortices were divided in six treatment groups: 1) 10 ng/mL EGF (EGF V0); 2) 10 ng/mL of EGF plus 1 mM Z-VAD-FMK (EGF V1); 3) 10 ng/mL of EGF and 10 mM Z-VAD-FMK (EGF V10); 4) 1 mM Z-VAD-FMK; 5) 10 mM Z-VAD-FMK and (6) no EGF and Z-VAD-FMK supplementation. Ovarian follicles within the tissues were processed for histology and assessed for follicle density, viability (based on morphology) and diameter immediately after collection (Control) or after 3 or 7 days of in vitro incubation. Comparison among fresh and culture treatment group was performed using ANOVA test. There were no differences (P > 0.05) in follicle density and viability among different culture treatments. However, there were differences in this parameter between culture days. Specifically, culturing tissue for 7 days resulted in significant reduction in follicle viability and density, regardless of treatments. We found a difference in size between culture days when these follicles were cultured using 10 mM Z-VAD-FMK or 10 ng/mL EGF (EGF V0). In sum, the finding demonstrated that Z-VAD-FMK at the dosage used in the present study does not provide the protective effect to ovarian tissue during in vitro culture. Future studies should explore different Z-VAD-FMK dosages or other anti-apoptotic agent, such as surviving in protecting ovarian follicles against cell death.

Keywords: anti apoptotic drug, bitches, follicles, Z-VAD-FMK

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