Search results for: neem leaf protein

Authors: Hien Thi Thu Le, Nuno Rafael Candeias, Olli Yli-Harja, Meenakshisundaram Kandhavelu

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Purinergic receptor 1 (P2Y1R) is the potential therapeutic target for inducing prostate cancer (PCa) cell death. Recently, 1-indolinoalkyl 2-phenolic derivative, HIC, was identified as a P2Y1R agonist that increases apoptosis and inhibits cell proliferation of PCa. However, the biological effects of HIC have not been extensively studied at the molecular level. In the present study, we have investigated the anticancer effects of HIC and the molecular mechanisms underlying in PCa cells. Half maximal inhibitory concentration (IC₅₀) of HIC was measured as 15.98 μM and 15.64 μM for DU145 and PC3 cells, respectively. In addition, we found that HIC inhibited cell growth and metastasis of PC3 and DU145 cells colonies, spheroid areas, and migrated cells. RNA seq analysis revealed significant changes of over 3000 genes (p value < 0.05) upon HIC treatment in PC3 and DU145 cells. Genes involved in DNA damage, apoptosis, cell cycle arrest at G1/S phase were modulated by HIC treatment. MAPK and NF-κB protein array revealed the increased expression of ERK1/2, JNK1/2, p53 phosphorylation, and p53 protein. ERK1/2 and JNK1/2 activations are known to increase the stabilization of p53, a tumor suppressor protein, which is required to arrest the cell cycle at G1/S phase and cause cell death of PCa cells. Overall, our results suggest that HIC can serve as a multi-dimensional chemotherapeutic agent possessing strong cytotoxic, anti-cancer, and anti-metastasis against PCa growth.

Keywords: prostate cancer, P2Y1 receptor, apoptosis, metastasis

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Authors: Manal Kamel, Sara Maher, Hanan El Baz, Faten Salah, Omar Sayyouh, Zeinab Demerdash

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In the recent COVID 19 pandemic, experts of public health have emphasized testing, tracking infected people, and tracing their contacts as an effective strategy to reduce the spread of the virus. Development of rapid and sensitive diagnostic assays to replace reverse transcription polymerase chain reaction (RT-PCR) is mandatory..Our innovative test strip relying on the application of nanoparticles conjugated to recombinant nanobodies for SARS-COV-2 spike protein (S1) & angiotensin-converting enzyme 2 (that is responsible for the virus entry into host cells) for rapid detection of SARS-COV-2 spike protein (S1) in saliva or sputum specimens. Comparative tests with RT-PCR will be held to estimate the significant effect of using COVID 19 nanobodies for the first time in the development of lateral flow test strip. The SARS-CoV-2 S1 (3 ng of recombinant proteins) was detected by our developed LFIA in saliva specimen of COVID-19 Patients No cross-reaction was detected with Middle East respiratory syndrome coronavirus (MERS-CoV) or SARS- CoV antigens..Our developed system revealed 96 % sensitivity and 100% specificity for saliva samples compared to 89 % and 100% sensitivity and specificity for nasopharyngeal swabs. providing a reliable alternative for the painful and uncomfortable nasopharyngeal swab process and the complexes, time consuming PCR test. An increase in testing compliances to be expected.

Keywords: COVID 19, diagnosis, LFIA, nanobodies, ACE2

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Authors: Rana Bilal, Faiz-Ul-Hassan, Moazzam Jameel

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There is a dire need to replace high-cost protein with more economical protein to overcome animal protein shortage in developing nations especially countries like Pakistan. In conjunction with these efforts, the current study was planned to evaluate the effects of various dried distillery yeast sludge (DYS) levels on the immune level, egg quality, and performance of layers by replacing soybean meal. The study was designed with two hundred layers of Hy-Line variety. Distillery yeast sludge was dried and ground for 2 mm mesh size and after this proximate and mineral analysis was determined. Five isocaloric and isonitrogeneous feeds were given containing C (control), 5, 10, 15, 20% distillery yeast sludge by replacing soybean meal. The trial was performed in the completely randomized design with five treatments, 4 replicates and 10 hen per replicate. Results demonstrated that feed intake, egg production, feed conversion ratio decreased (P < 0.05) with the increased dietary DYS. However, statistically significant decrease (P < 0.05) was found in hens having DYS20 diet than control. Layers on Diets C, DYS5 and DYS10 exerted a higher immune level than DYS15 and DYS20 diets. Egg weight, eggshell weight, eggshell thickness, egg albumen height as well as haugh unit score were affected significantly by the increased level of DYS. In general, results of this study demonstrated that inclusion of DYS up to 10% showed no adverse effects on health and performance of layers.

Keywords: egg quality, immunity, layers, performance

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Authors: Shalini Singh, Sanjay K. Srivastava, Govind, Mukhtar. A. Khan, P. K. Singh

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Nanotechnology is revolutionizing the development of biosensors. Nanomaterials and nanofabrication technologies are increasingly being used to design novel biosensors. Sensitivity and other attributes of biosensors can be improved by using nanomaterials with unique chemical, physical, and mechanical properties in their construction. Silicon is a promising biomaterial that is non-toxic and biodegradable and can be exploited in chemical and biological sensing. Present study demonstrated the streptavidin–biotin interaction on silicon surfaces with different topographies such as flat and nanostructured silicon (nanowires) surfaces. Silicon nanowires with wide range of surface to volume ratio were prepared by electrochemical etching of silicon wafer. The large specific surface of silicon nanowires can be chemically modified to link different molecular probes (DNA strands, enzymes, proteins and so on), which recognize the target analytes, in order to enhance the selectivity and specificity of the sensor device. The interaction of streptavidin with biotin was carried out on 3-aminopropyltriethoxysilane (APTS) functionalized silicon surfaces. Fourier Transform Infrared Spectroscopy (FTIR) and X-ray Photoelectron Spectroscopy (XPS) studies have been performed to characterize the surface characteristics to ensure the protein attachment. Silicon nanowires showed the enhance protein attachment, as compared to flat silicon surface due to its large surface area and good molecular penetration to its surface. The methodology developed herein could be generalized to a wide range of protein-ligand interactions, since it is relatively easy to conjugate biotin with diverse biomolecules such as antibodies, enzymes, peptides, and nucleotides.

Keywords: FTIR, silicon nanowires, streptavidin-biotin, XPS

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Authors: Solomon Assefa Huluka

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Wounds account for severe morbidity, socioeconomic distress, and mortality around the globe.For several years, various herbal products are used to expediteand augment the innate wound healing process. In Ethiopian folkloricmedicine, Verbascum sinaiticum L. (V. sinaiticum) is commonlyapplied as a wound-healing agent. The present study investigated the potential wound healing and antioxidant properties of hydroalcoholic leaf extract of V. sinaiticum. The 80% methanol extract, formulated as 5% (w/w) and 10% (w/w) ointments, was evaluated in excision and incision wound models using nitrofurazone and simple ointment as positive and negative controls, respectively. Parameters such as wound contraction, period of epithelialization, and tensile strength were determined. Moreover, its in vitro antioxidant property was evaluated using a DPPH assay. In the excision model, both doses (5% and 10% w/w) of the extract showed a significant (p<0.001) wound healing efficacy compared to the negative control, as evidenced by enhanced wound contraction rate and shorter epithelialization time records. In the incision model, the lower dose (5% w/w) ointment formulation of the extract exhibited the maximum increment in tensile strength (85.6%) that was significant (p<0.001)compared to negative and untreated controls. Animals treated with 5% w/wointment, furthermore, showed a significantly (p < 0.05) higher percentage of tensile strength than nitrofurazone treated ones. Moreover, the hydroalcoholic extract of the plant showed a noticeable free radical scavenging property. The result of the present study upholds the folkloric use of V. sinaiticum in the treatment of wounds.

Keywords: wound healing, antioxidant, excision wound model, incision wound model, verbascum sinaiticum

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Authors: M. Dehestani, F. Kamali, M. Klantari Pour, L. Zeidabadi-Nejad

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Chemical bonding between polyethylene glycol (PEG) with pharmaceutical proteins called pegylation is one of the most effective methods of improving the pharmacological properties. The covalent attachment of polyethylene glycol (PEG) to proteins will increase their pharmacologic properties. For the formation of a combination of pegylated protein should first be activated PEG and connected to the protein. Interferons(IFNs) are a family of cytokines which show antiviral effects in front of the biological and are responsible for setting safety system. In this study, the nature and properties of the interaction between active positions of IFNs and polyethylene glycol have been investigated using molecular dynamics simulation. The main aspect of this theoretical work focuses on the achievement of valuable data on the reaction pathways of PEG-IFNs and the transition state energy. Our results provide a new perspective on the interactions, chemical properties and reaction pathways between IFNs and PEG.

Keywords: interaction, interferons, molecular dynamics simulation, polyethylene glycol

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Authors: Muhammad Saeed, Nazeer Ahmed, Mukhtar Alam, Fazli Subhan, Muhammad Adnan, Fazli Wahid, Hidayat Ullah, Rafiullah

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The present experiment evaluated different synthetic insecticides against Jassid (Amrasca devastations) on pea crop at Agriculture Research Institute Tarnab, Peshawar Khyber Pakhtunkhwa. The field was prepared to cultivate okra crop in Randomized Complete Block (RCB) Design having six treatments with four replications. Plant to plant and row to row distance was kept at 15 cm and 30 cm, respectively. Pre and post spray data were recorded randomly from the top, middle and bottom leaves of five selected plants. Five synthetic insecticides, namely Confidor (Proponil), a neonicotinoid insecticide, Chlorpyrifos (chlorinated organophosphate (OP) insecticide), Lazer (dinitroaniline) (Pendimethaline), Imidacloprid (neonicotinoids insecticide) and Thiodan (Endosulfan, organochlorine insecticide), were used against infestation of aphids, pea pod borer, stem fly, leaf minor and pea weevil. Each synthetic insecticide showed significantly more effectiveness than control (untreated plots) but was non-significant among each other. The lowest population density was recorded in the plot treated with synthetic insecticide i.e. Confidor (0.6175 liter.ha-1) (4.24 aphids plant⁻¹) which is followed by Imidacloprid (0.6175 liter.ha⁻¹) (4.64 pea pod borer plant⁻¹), Thiodan (1.729 liter.ha⁻¹) (4.78 leaf minor plant⁻¹), Lazer (2.47 liter.ha-1) (4.91 pea weevil plant⁻¹), Chlorpyrifos (1.86 liter.ha⁻¹) (5.11 stem fly plant⁻¹), respectively while the highest population was recorded from the control plot. It is concluded from the data that the residual effect decreases with time after the application of spray, which may be less dangerous to the environment and human beings and can effectively manage this dread.

Keywords: okra crop, jassids, Confidor, imidacloprid, chlorpyrifos, laser, Thiodan

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Authors: Sujata Sharma, Avinash Singh, Lovely Gautam, Pradeep Sharma, Mau Sinha, Asha Bhushan, Punit Kaur, Tej P. Singh

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Peptidyl-tRNA hydrolase (Pth) Pth is an essential bacterial enzyme that catalyzes the release of free tRNA and peptide moeities from peptidyl tRNAs during stalling of protein synthesis. In order to design inhibitors of Pth from Pseudomonas aeruginosa (PaPth), we have determined the structures of PaPth in its native state and in the bound states with two compounds, amino acylate-tRNA analogue (AAtA) and 5-azacytidine (AZAC). The peptidyl-tRNA hydrolase gene from Pseudomonas aeruginosa was amplified by Phusion High-Fidelity DNA Polymerase using forward and reverse primers, respectively. The E. coliBL21 (λDE3) strain was used for expression of the recombinant peptidyl-tRNA hydrolase from Pseudomonas aeruginosa. The protein was purified using a Ni-NTA superflow column. The crystallization experiments were carried out using hanging drop vapour diffusion method. The crystals diffracted to 1.50 Å resolution. The data were processed using HKL-2000. The polypeptide chain of PaPth consists of 194 amino acid residues from Met1 to Ala194. The centrally located β-structure is surrounded by α-helices from all sides except the side that has entrance to the substrate binding site. The structures of the complexes of PaPth with AAtA and AZAC showed the ligands bound to PaPth in the substrate binding cleft and interacted with protein atoms extensively. The residues that formed intermolecular hydrogen bonds with the atoms of AAtA included Asn12, His22, Asn70, Gly113, Asn116, Ser148, and Glu161 of the symmetry related molecule. The amino acids that were involved in hydrogen bonded interactions in case of AZAC included, His22, Gly113, Asn116, and Ser148. As indicated by fittings of two ligands and the number of interactions made by them with protein atoms, AAtA appears to be a more compatible with the structure of the substrate binding cleft. However, there is a further scope to achieve a better stacking than that of O-tyrosyl moiety because it is not still ideally stacked. These observations about the interactions between the protein and ligands have provided the information about the mode of binding of ligands, nature and number of interactions. This information may be useful for the design of tight inhibitors of Pth enzymes.

Keywords: peptidyl tRNA hydrolase, Acinetobacter baumannii, Pth enzymes, O-tyrosyl

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Authors: Taslima Zahan

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A field study was done in 2013-14 and 2014-15 by following bio-assay technique to determine the carryover effect of herbicides applied in rainy season rice on growth and yield of two probable succeeding crops of rice viz., wheat and lentil. Rice seedlings were transplanted on strip-tilled non-puddled field, and five herbicides named pyrazosufuron-ethyl, butachlor, orthosulfamuron, butachlor + propanil and 2,4-D amine were applied in rice at their recommended rate and time as eight treatment combinations and compared with one untreated control. Residual effects of those rice herbicides on the succeeding wheat and lentil were examined by following micro-plot bioassay technique. The study revealed that germination of wheat and lentil seeds were not affected by the residue of herbicides applied in the preceding rainy season rice. Shoot length of wheat and lentil seedlings of herbicide treated plots were also non-significantly varied with untreated control plots. Herbicide treated plots of wheat had higher leaf chlorophyll contents over the control plots by 1.8-14.0% on an average while in case of lentil herbicide treated plots had negligible amount of reduction in leaf chlorophyll contents than control plots. Grain yields of wheat and lentil in herbicide treated plots were higher than control plots by 2.8-6.6% and 0.2-10.9%, respectively. Therefore, two-year bioassay study claimed that tested herbicides applied in rainy season rice under strip-tilled non-puddled field had no adverse residual effect on growth and yield of the succeeding wheat and lentil.

Keywords: crop sensitivity, herbicide persistence, minimum tillage rice, yield improvement

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Authors: Lebogang L. R. Mphahlele, Bruce B. Sithole

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Chicken meat is the highest consumed meat in south Africa, with a per capita consumption of >33 kg yearly. Hence, South Africa produces over 250 million kg of waste chicken feathers each year, the majority of which is landfilled or incinerated. The discarded feathers have caused environmental pollution and natural protein resource waste. Therefore, the valorisation of waste chicken feathers is measured as a more environmentally friendly and cost-effective treatment. Feather contains 91% protein, the main component being beta-keratin, a fibrous and insoluble structural protein extensively cross linked by disulfide bonds. Keratin is usually converted it into nanofibers via electrospinning for a variety of applications. keratin nanofiber composites have many potential biomedical applications for their attractive features, such as high surface-to-volume ratio and very high porosity. The application of nanofibers in the biomedical wound dressing requires antimicrobial properties for materials. One approach is incorporating inorganic nanoparticles, among which silver nanoparticles played an important alternative antibacterial agent and have been studied against many types of microbes. The objective of this study is to combine synthetic polymer, chicken feather keratin, and antibacterial nanoparticles to develop novel electrospun antibacterial nanofibrous composites for possible wound dressing application. Furthermore, this study will converting a two-dimensional electrospun nanofiber membrane to three-dimensional fiber networks that resemble the structure of the extracellular matrix (ECM)

Keywords: chicken feather keratin, nanofibers, nanoparticles, nanocomposites, wound dressing

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Authors: Hanaa A. S. Oraby, Amal A. M. Hassan, Mahmoud M. Sakr, Atef A. A. Haiba

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Applying the profiling approach for the assessment of proteomic variations due to genetic modification of the Egyptian tomato cultivar "Edkawy", three complementary approaches were used. These methods are amino acids analysis, gel electrophoresis, and Gas chromatography coupled with mass spectrometry (GC/MS). The results of the present study Show evidence of proteomic variations between both modified tomato and its non-modified counterpart. Amino acids concentrations, and the protein patterns separation on the 1D SDS-PAGE were not similar in the case of transformed tomato compared to that of the non-transformed counterpart. These detected differences are most likely derived from the process of transformation. Results also revealed that the efficiency of GC/MS approach to identify a mixture of unknown proteins is limited. GC/MS analysis was only able to identify few number of protein molecules. Therefore, more advanced and specific technologies like MALDI-TOF-MS are recommended to be employed.

Keywords: GMOs, unintended effects, proteomic variations, 1D SDS-PAGE, GC/MS

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Authors: M. Cherki, I. Taouam, A. Amiri, F. Hmimid, T. Ould Bellahcen

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The Moroccan coasts represent an important wealth of red algae which have an economic interest. Among these algae, the Gelidium sesquipedale, which is exploited industrially for its richness in agar. The aim of this study is to establish a general overview of the macronutrient composition of Gelidium sesquipedale and to compare this composition according to three factors: the harvest site (El Jadida, Casablanca and Mohammadia), the harvest depth (coast and depth) and the drying process (open air and oven). Proteins, lipids, and carbohydrates are measured by different methods. The analysis of results show that the protein concentrations of the El Jadida and Mohammadia samples are significantly higher than that of Casablanca (0.026 ± 0.0007 µg/µg DW 0.024 ± 0.001 µg/µg DW and 0.006 ± 0.0007 µg/µg DW, p < 0.05 respectively). However, Casablanca samples are significantly richer in total sugars (0.023 ± 0.002 µg/µg DW, p < 0.05) and less rich in reducing sugars (0.0001 ± 0.00001 µg/µg DW, p < 0.05) compared to other samples. The lipid concentrations of the samples from the three harvest sites do not show any significant difference. With respect to depth, only total protein and total sugar concentrations were significantly higher in the coast versus depth samples (0.035 ± 0.004 µg/µg DW vs. 0.026 ± 0.0007 µg/µg DW and 0.035 ± 0.006 µg/µg DW vs. 0.012 ± 0.005 µg/µg DW p < 0.05 respectively). For the drying process, protein, total sugars and lipid concentrations were significantly higher in open air samples compared to oven samples (0.006 ± 0.0007 µg/µg DW). vs 0.004 ± 0.0003 µg/µg DW, 0.023 ± 0.002 µg/µg DW vs 0.007 ± 0.002 µg/µg DW and 8% vs 4% p < 0.05 respectively). Our results demonstrate that the chemical composition of Gelidium sesquipedale varies according to the harvest region. In addition, samples harvested on the coast and dried in the open air are the richest in macronutrients.

Keywords: biochemical composition, drying, depth, Gelidium sesquipedale, red algae, region

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Authors: Rashmi Ranade, Neelu Joshi

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Barleria prionitis L. is a well explored Indian medicinal plant valued for its stem and leaf which forms an important ingredient of many Ayurvedic formulations. It is used for the treatment of various disorders like toothache, bleeding gums, strengthening gums, whooping cough, inflammation, arthritis, enlargement of scrotum and sciatica etc. The plant is propagated vegetatively through stem cuttings. Frequent harvesting of this plant has led to the shortage of planting material, and it has acquired the status of vulnerable plant species. Plant tissue culture technology offers a very good alternative for propagation and conservation of such plant species. The present investigation was undertaken to develop in vitro regeneration protocol for B. prionitis L. via callus organogenesis pathway. Stem and leaf explants were used for this purpose. Different media and plant growth regulators were optimized to develop the protocol. The problem of phenol secretion and browning and in vitro cultures at the establishment phase was successfully curbed with the usage of antibrowning agents such as ascorbic acid and activated charcoal. Optimum shoot multiplication was achieved by the use of liquid media and incorporation of silver nitrate and TIBA (triiodobenzoic acid) into the media. High percent rooting (76%) was observed on WPM media supplemented with IBA (2.0 mg/l), IAA (0.5 mg/l), GA3(0.5) and activated charcoal(500 mg/l). The rooted plantlets were subjected to in vitro hardening on sterile potting mix (soil:farmyard manure:compost; 1:2:1) and acclimatized under greenhouse conditions. Around 85% survival of plantlets was recorded upon acclimatization. This lab scale protocol would be tested for in vitro scaling up production of B. prionitis L.

Keywords: explant browning, liquid culture, micropropagation, shoot multiplication, phenolic secretion

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Authors: Kai Pai Huang

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Background: There are several types of plasma cell neoplasms including multiple myeloma, plasmacytoma, lymphoplasmacytic lymphoma, and monoclonal gammopathy of undetermined significance (MGUS) are found in our lab. Today, we want to compare with two cases using the method of capillary electrophoresis. Method: Serum is prepared and electrophoresis is performed at alkaline PH in a capillary using the Sebia® Capillary 2. Albumin and globulins are detected by the detector which is located in the cathode of the capillary and the signals are transformed to peaks. Serum was treated with beta-mercaptoethanol which reducing the polymerized immunoglobulin to monomer immunoglobulin to clarify two M-protein are secreted from the same plasma cell clone in bone marrow. Result: Case 1: A 78-year-old female presenting dysuria, oliguria and leg edema for several months. Laboratory data showed proteinuria, leukocytosis, results of high serum IgA and lambda light chain. A renal biopsy found amyloid fibrils in the glomerular mesangial area. Serum protein electrophoresis shows a major monoclonal peak in the β region and minor small peak in gamma region, and the immunotyping studies for serum showed two IgA/λ type. Case 2: A 55-year-old male presenting abdominal distension and low back pain for more than one month. Laboratory data showed T12 T8 compression fracture, results of high serum IgM and kappa light chain. Bone marrow aspiration showed the cells from the bone marrow are B cells with monotypic kappa chain expression. Bone marrow biopsy found this is lymphoplasmacytic lymphoma (Waldenstrom macroglobulin). Serum protein electrophoresis shows a monoclonal peak in the β region and the immunotyping studies for serum showed IgM/κ type. Conclusion: Plasma cell neoplasm can be diagnosed by many examinations. Among them, using capillary electrophoresis by a lab can separate several types of gammopathy and the quantification of a monoclonal peak can be used to evaluate the patients’ prognosis or treatment.

Keywords: plasma cell neoplasm, capillary electrophoresis, serum protein electrophoresis, immunotyping

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Authors: Sulhee Lee, Geon Kim, Young-Seo Park

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Unlike the traditional milk sterilization methods (LTLT, HTST, or UHT), pulsed electric field (PEF) technology is a non-thermal pasteurization process. This technology minimizes energy required for heat treatment in food processing, changes in sensory properties, and physical losses. In this study, structural changes of bovine milk proteins, the amount of immunoproteins such as IgG, and their storability by PEF treatment were examined. When the changes of protein content in PEF-treated milk were examined using HPLC, the amounts of α-casein and β-lactoglobulin were reduced over 40% each, whereas those of κ-casein and β-casein did not change. The amount of α-casein in HTST milk was reduced to 50%. When PEF was applied to milk at the energy level of 250 kJ, the amounts of IgG, IgA, β-lactoglobulin (β-LG), lactoferrin, and α-lactalbumin (α-LA) decreased by 43, 41, 35, 63, and 45%, respectively. When milk was sterilized by LTLT process followed by PEF process at the level of 150 kJ, the concentrations of IgG, IgA, β-LG, lactoferrin, and α-LA were 56.6, 10.6, 554, 2.8 and 660.1 μg/mL, respectively. When the bovine milk was sterilized by LTLT process followed by PEF process at the energy level of 180 kJ, storability of immunoproteins of milk was the highest and the concentrations of IgG, IgA, and β-LG decreased by 79.5, 6.5, and 134.5 μg/mL, respectively, when compared with the initial concentrations of those proteins. When bovine milk was stored at 4℃ after sterilization through HTST sterilizer followed by PEF process at the energy level of 200 kJ, the amount of lactoferrin decreased 7.3% after 36 days of storage, whereas that of lactoferrin of raw milk decreased 16.4%. Our results showed that PEF treatment did not change the protein structure nor induce protein denaturation in milk significantly when compared with LTLT or HTST sterilization. Also, LTLT or HTST process in combination with PEF were more effective than LTLT only or HTST only process in the conservation of immunoproteins in bovine milk.

Keywords: pulsed electric field, bovine milk, immunoproteins, sterilization

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Authors: Januar Parlaungan Siregar, Davindra Brabu Mathivanan, Dandi Bachtiar, Mohd Ruzaimi Mat Rejab, Tezara Cionita

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Natural fibres play a significant role in mass industries such as automotive, construction and sports. Many researchers have found that the natural fibres are the best replacement for the synthetic fibres in terms of cost, safety, and degradability due to the shortage of landfill and ingestion of non biodegradable plastic by animals. This study mainly revolved around pineapple leaf fibre (PALF) which is available abundantly in tropical countries and with excellent mechanical properties. The composite formed in this study is highly biodegradable as both fibre and matrix are both derived from natural based products. The matrix which is polylactic acid (PLA) is made from corn starch which gives the upper hand as both material are renewable resources are easier to degrade by bacteria or enzyme. The PALF is treated with different alkaline solution to remove excessive moisture in the fibre to provide better interfacial bonding with PLA. Thereafter the PALF is washed with distilled water several times before placing in vacuum oven at 80°C for 48 hours. The dried PALF later were mixed with PLA using extrusion method using fibre in percentage of 30 by weight. The temperature for all zone were maintained at 160°C with the screw speed of 50 rpm for better bonding and afterwards the products of the mixture were pelletized using pelletizer. The pellets were placed in the specimen-sized mould for hot compression under the temperature of 170°C at 5 MPa for 5 min and subsequently were cold pressed under room temperature at 5 MPa for 5 min. The specimen were tested for tensile and flexure strength according to American Society for Testing and Materials (ASTM) D638 and D790 respectively. The effect of surface modification on PALF with different alkali solution will be investigated and compared.

Keywords: natural fibre, PALF, PLA, composite

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Authors: J. Dharmaraj, C. Gunasekaran

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Invertebrates are the reliable ecological indicators of disturbance of the forest ecosystems and they respond to environment changes more quickly than other fauna. Among these the terrestrial invertebrates are vital to functioning ecosystems, contributing to processes such as decomposition, nutrient cycling and soil fertility. The natural ecosystems of the forests have been subject to various types of disturbances, which lead to decline of flora and fauna. The comparative diversity of micro-arthropods in natural forest, wattle plantation and eucalyptus plantations were studied in Nilgiris. The study area was divided in to five major sites (Emerald (Site-I), Thalaikundha (Site-II), Kodapmund (Site-III), Aravankad (Site-IV), Kattabettu (Site-V). The research was conducted during period from March 2014 to August 2014. The leaf and soil samples were collected and isolated by using Berlese funnel extraction methods. Specimens were isolated and identified according to their morphology (Balogh 1972). In the present study results clearly showed the variation in soil pH, NPK (Major Nutrients) and organic carbon among the study sites. The chemical components of the leaf litters of the plantation decreased the diversity of micro-arthropods and decomposition rate leads to low amount of carbon and other nutrients present in the soil. Moreover eucalyptus and wattle plantations decreases the availability of the ground water source to other plantations and micro-arthropods and hences affects the soil fertility. Hence, the present study suggests to minimize the growth of wattle and eucalyptus tree plantations in the natural areas which may help to reduce the decline of forests.

Keywords: micro-arthropods, assemblage, berlese funnel, morphology, NPK, nilgiris

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Authors: Aminah Muchdar

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To increase the genetic diversity of soybean in order to adapt to agroecology in Indonesia conducted ways including introduction, cross, mutation and genetic transformation. The purpose of this research is to obtain early maturity soybean mutant lines, large seed tolerant to drought with high yield potential. This study consisted of two stages: the first is sensitivity of gamma rays carried out in the Laboratory BATAN. The genetic variety used is Anjasmoro. The method seeds irradiated with gamma rays at a rate of activity with the old ci 1046.16976 irradiation 0-71 minutes. Irradiation doses of 0, 100, 200, 300, 400, 500, 600, 700, 800, 900 and 1000gy. The results indicated all seeds irradiated with doses of 0 - 1000gy, just a dose of 200 and 300gy are able to show the percentage of germination, plant height, number of leaves, number of normal sprouts and green leaves of the best and can be continued for a second trial in order to assemble and to get mutants which is expected. The result of second stage of soybean M2 Population irradiated with diversity Gamma Irradiation performed that in the form of soybean planting, the seed planted is the first derivative of the M2 irradiated seeds. The result after the age of 30ADP has already showing growth and development of plants that vary when compared to its parent, both in terms of plant height, number of leaves, leaf shape and leaf forage level. In the generative phase, a plant that has been irradiated 200 and 300 gy seen some plants flower form packs, but not formed pods, there is also a form packs of flowers, but few pods produce soybean morphological characters such as plant height, number of branches, pods, days to flowering, harvesting, seed weight and seed number.

Keywords: gamma ray, genetic mutation, irradiation, soybean

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Authors: Elif Gencturk, Ekin Yurdakul, Ahmet Y. Celik, Senol Mutlu, Kutlu O. Ulgen

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Yeast cells are generally used as a model system of eukaryotes due to their complex genetic structure, rapid growth ability in optimum conditions, easy replication and well-defined genetic system properties. Thus, yeast cells increased the knowledge of the principal pathways in humans. During fermentation, carbohydrates (hexoses and pentoses) degrade into some toxic by-products such as 5-hydroxymethylfurfural (5-HMF or HMF) and furfural. HMF influences the ethanol yield, and ethanol productivity; it interferes with microbial growth and is considered as a potent inhibitor of bioethanol production. In this study, yeast single cell behavior under HMF application was monitored by using a continuous flow single phase microfluidic platform. Microfluidic device in operation is fabricated by hot embossing and thermo-compression techniques from cyclo-olefin polymer (COP). COP is biocompatible, transparent and rigid material and it is suitable for observing fluorescence of cells considering its low auto-fluorescence characteristic. The response of yeast cells was recorded through Red Fluorescent Protein (RFP) tagged Nop56 gene product, which is an essential evolutionary-conserved nucleolar protein, and also a member of the box C/D snoRNP complexes. With the application of HMF, yeast cell proliferation continued but HMF slowed down the cell growth, and after HMF treatment the cell proliferation stopped. By the addition of fresh nutrient medium, the yeast cells recovered after 6 hours of HMF exposure. Thus, HMF application suppresses normal functioning of cell cycle but it does not cause cells to die. The monitoring of Nop56 expression phases of the individual cells shed light on the protein and ribosome synthesis cycles along with their link to growth. Further computational study revealed that the mechanisms underlying the inhibitory or inductive effects of HMF on growth are enriched in functional categories of protein degradation, protein processing, DNA repair and multidrug resistance. The present microfluidic device can successfully be used for studying the effects of inhibitory agents on growth by single cell tracking, thus capturing cell to cell variations. By metabolic engineering techniques, engineered strains can be developed, and the metabolic network of the microorganism can thus be manipulated such that chemical overproduction of target metabolite is achieved along with the maximum growth/biomass yield.  

Keywords: COP, HMF, ribosome biogenesis, thermoplastic microbioreactor, yeast

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Authors: Hainan Chen, Jina Qing, Xiao Zhu, Ling Gao, Ampadu O. Jackson, Min Zhang, Kai Yin

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Objective: Editing macrophage activation to dampen inflammatory diseases by promoting the repolarization of inflammatory (M1) macrophages to anti-inflammatory (M2) macrophages is highly associated with mitochondrial respiration. Recent studies have suggested that mitochondrial apolipoprotein A-1 binding protein (APOA1BP) was essential for the cellular metabolite NADHX repair to NADH, which is necessary for the mitochondrial function. The exact role of APOA1BP in the repolarization of M1 to M2, however, is uncertain. Material and method: THP-1-derived macrophages were incubated with LPS (10 ng/ml) or/and IL-4 (100 U/ml) for 24 hours. Biochemical parameters of oxidative phosphorylation and M1/M2 markers were analyzed after overexpression of APOA1BP in cells. Results: Compared with control and IL-4-exposed M2 cells, APOA1BP was downregulated in M1 macrophages. APOA1BP restored the decline in mitochondrial function to improve metabolic and phenotypic reprogramming of M1 to M2 macrophages. Blocking oxidative phosphorylation by oligomycin blunts the effects of APOA1BP on M1 to M2 repolarization. Mechanistically, LPS triggered the hydration of NADH and increased its hydrate NADHX which inhibit cellular NADH dehydrogenases, a key component of electron transport chain for oxidative phosphorylation. APOA1BP decreased the level of NADHX via converting R-NADHX to biologically useful S-NADHX. The mutant of APOA1BP aspartate188, the binding site of NADHX, fail to repair oxidative phosphorylation, thereby preventing repolarization. Conclusions: Restoring mitochondrial function by increasing mitochondrial APOA1BP might be useful to improve the reprogramming of inflammatory macrophages into anti-inflammatory cells to control inflammatory diseases.

Keywords: inflammatory diseases, macrophage repolarization, mitochondrial respiration, apolipoprotein A-1 binding protein, NADHX, NADH

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Authors: Bendjazia Radia, Samira Kilani-Morakchi, Nadia Aribi

Abstract:

Azadirachtin is a biorational insecticide commonly reported as selective to a range of beneficial insects. It is one of the most biologically active natural inhibitors of insect growth and development and it is known to be an antagonist of the juvenile hormone and 20-hydroxyecdysone (20E). However, its mechanism of action remains still unknown. In the present study, the toxicity of a commercial formulation of Azadirachtin (Neem Azal, 1% azadirachtine) was evaluated by topical application at various doses (0.1, 0.25, 0.5, 1 and 2 µg/insect) on the third instars larvae of D. melanogaster. Lethal doses (LD25: 0.28µg and LD50: 0.67µg), were evaluated by cumulated mortality at the immature stages. The effects of azadirachtin (LD25 and LD50) were then evaluated on the development (duration of the larval and pupal instars, the weight of larvae, pupa and adults) of Drosophila melanogaster. Results showed that the insecticide increased significantly the larval and pupal instar duration. A reduction of larval and pupal weight is noted under azadirachtin treatment as compared to controls. In addition, the weight of surviving adults at the two tested dose was also reduced. In conclusion, azadirachtin seemed to interfere with the functions of the endocrine system resulting in development defects.

Keywords: azadirachtin, d.melanogaster, toxicity, development

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Authors: Zeiler M., Stadler D., Schmaderer C.

Abstract:

Over the years of dialysis treatment, most patients experience significant weight loss. The primary emphasis in earlier research was the underlying mechanism of protein energy wasting and the subsequent malnutrition inflammation syndrome. In the interest to provide an effective and rapid solution for the patients, the aim of this study is identifying individual influences of their assumed reduced dietary intake, such as nausea, appetite loss and taste changes, and to determine whether the patients adhere to their nutrition guidelines. A prospective, controlled study with 38 end-stage renal disease patients was performed using a questionnaire to reflect their diet within the last 12 months. Thereby, the daily intake for the most important macro-and micronutrients was calculated to be compared with the individual KDQOI-guideline value, as well as controls matched in age and gender. The majority of the study population did not report symptoms commonly associated with dialysis, such as nausea or inappetence, and denied any change in dietary behavior since receiving renal replacement therapy. The patients’ daily intake of energy (3080kcal ± 1266) and protein (89,9g [53,4-142,0]) did not differ significantly from the controls (energy intake: 3233kcal ± 1046, p=0,597; protein intake: 103,7g [90,1-125,5], p=0,120). The average difference to the individual calculated KDQOI-guideline was +176,0kcal ± 1156 (p=0,357) for energy intake and -1,75g ± 45,9 (p=0,491) for protein intake. However, there was an observed imbalance in the distribution of macronutrients, with a preference for fats over proteins. The patients’ daily intake of sodium (5,4g [ 2,95-10,1]) was higher than in the controls (4,1g [2,04-5,99], p= 0,058) whereas both values for potassium (3,7g ± 1,84) and phosphorous (1,79g ± 0,91) went significantly below the controls’ values (potassium intake: 4,89g ± 1,74, p=0,014; phosphorous intake: 2,04g ± 0,64, p=0,038). Thus, the values exceeded the calculated KDQOI-recommendation by + 3,3g [0,63-7,90] (p<0,001) for sodium, +1,49g ± 1,84 (p<0,001) for potassium and +0,89g ± 0,91 (p<0,001) for phosphorous. Contrary to the assumption, the patients did not under-eat. Nevertheless, their diets did not align with the recommended values. These findings highlight the need for intervention and education among patients and that regular dietary monitoring could prevent unhealthy nutrition habits. The elaboration of individual references instead of standardized guidelines could increase the compliance to the advised diet so that interdisciplinary comorbidities do not develop or worsen.

Keywords: compliance, dialysis, end-stage renal disease, KDQOI, malnutrition, nutrition guidelines, questionnaire, salt intake

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Authors: Ajith K. A. Gedara, Iva Chianella, Jose L. Endrino, Qi Zhang

Abstract:

The olive seed is a by-product of the olive oil production industry. Biuret test and ferric chloride test revealed that water or alkali NaOH extractions of olive seed flour are rich in proteins and tannins. Both protein and tannins are well-known bio-based wood adhesives in the wood-based panel industry. In general, tannins-based adhesives show better mechanical and physical properties than protein wood adhesives. This paper explores different methods of extracting tannins from olive seed flour against the tannins yield and their applications as bio-based adhesives in wood-based panels. Once investigated, the physical and the mechanical properties of wood-based panels made using bio-adhesives based tannins extracted from olive seed flour revealed that the resulting products seemed to satisfy the Japanese Industrial Standards JIS A 5908:2015.

Keywords: bio-adhesives, olive seed flour, tannins, wood-based panels

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Authors: J. Adeppa, S. Samanta, O. K. Raina

Abstract:

Fasciolosis is a widespread economically important parasitic infection throughout the world caused by Fasciola hepatica and F. gigantica. In order to identify novel immunogen conferring significant protection against fasciolosis, currently, research has been focused on the defined antigens viz. glutathione S-transferase, fatty acid binding protein, cathepsin-L, fluke hemoglobin, paramyosin, myosin and F. hepatica- Kunitz Type Molecule. Among various antigens, GST which plays a crucial role in detoxification processes, i.e. phase II defense mechanism of this parasite, has a unique position as a novel vaccine candidate and a drug target in the control of this disease. For producing the antigens in large quantities and their purification to complete homogeneity, the recombinant DNA technology has become an important tool to achieve this milestone. RT- PCR was carried out using F. gigantica total RNA as template, and an amplicon of 657 bp GST gene was obtained. TA cloning vector was used for cloning of this gene, and the presence of insert was confirmed by blue-white selection for recombinant colonies. Sequence analysis of the present isolate showed 99.1% sequence homology with the published sequence of the F. gigantica GST gene of cattle origin (accession no. AF112657), with six nucleotide changes at 72, 74, 423, 513, 549 and 627th bp found in the present isolate, causing an overall change of 4 amino acids. The 657 bp GST gene was cloned at BamH1 and HindIII restriction sites of the prokaryotic expression vector pPROEXHTb in frame with six histidine residues and expressed in E. coli DH5α. Recombinant protein was purified from the bacterial lysate under non-denaturing conditions by the process of sonication after lysozyme treatment and subjecting the soluble fraction of the bacterial lysate to Ni-NTA affinity chromatography. Western blotting with rabbit hyper-immune serum showed immuno-reactivity with 25 kDa recombinant GST. Recombinant protein detected F. gigantica experimental as well as field infection in buffaloes by dot-ELISA. However, cross-reactivity studies on Fasciola gigantica GST antigen are needed to evaluate the utility of this protein in the serodiagnosis of fasciolosis.

Keywords: fasciola gigantic, fasciola hepatica, GST, RT- PCR

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Authors: Sarah Elsayed Mohammed, Lamiaa Ahmed Ahmed, Mahmoud Mohammed Khattab

Abstract:

Aim: The aim of the present study was to investigate whether combined nicorandil and bone marrow-derived mesenchymal stem cells (BMDMSC) treatment could offer an additional benefit in ameliorating isoproterenol (ISO)-induced heart failure in rats. Methods: ISO (85 and 170 mg/kg/day) was injected subcutaneously for 2 successive days, respectively. By day 3, electrocardiographic changes were recorded and serum was separated for determination of CK-MB level for confirmation of myocardial damage. Nicorandil (3 mg/kg/day) was then given orally with or without a single i.v. BMDMSC administration. Electrocardiography and echocardiography were recorded 2 weeks after beginning of treatment. Rats were then sacrificed and ventricles were isolated for estimation of vascular endothelial growth factor (VEGF), tumor necrosis factor-alpha (TNF-α) and transforming growth factor-beta (TGF-β) contents, caspase-3 activity as well as inducible nitric oxide synthase (iNOS) and connexin-43 protein expressions. Moreover, histological analysis of myocardial fibrosis was performed and cryosections were done for estimation of homing of BMDMSC. Results: ISO induced a significant increase in ventricles/body weight ratio, left ventricular end diastolic (LVEDD) and systolic dimensions (LVESD), ST segment and QRS duration. Moreover, myocardial fibrosis as well as VEGF, TNF-α and TGF-β contents were significantly increased. On the other hand, connexin-43 protein expression was significantly decreased, while caspase-3 and iNOS protein expressions were significantly increased. Combined therapy provided additional improvement compared to cell treatment alone towards reducing cardiac hypertrophy, fibrosis and inflammation. Furthermore, combined therapy induced significant increase in angiogenesis and BMDMSC homing and prevented ISO induced changes in iNOS, connexin-43 and caspase-3 protein expressions. Conclusion: Combined nicorandil/BMDMSC treatment was superior to BMDMSC alone towards preventing ISO-induced heart failure in rats.

Keywords: fibrosis, isoproterenol, mesenchymal stem cells, nicorandil

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Authors: Nurcan Cetinkaya, Nadide Hulya Ozdemir

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The aim of the study was to investigate the effect of Saccharomyces cerevisiae (SC) live yeast culture on microbial protein supply to the small intestine in Kivircik male yearlings when fed with different ratio of forage and concentrate diets. Four Kivircik male yearlings with permanent rumen canula were used in the experiment. . The treatments were allocated to a 4x4 Latin square design. Diet I consisted of 70% alfalfa hay and 30% concentrate, Diet II consisted of 30% alfalfa hay and 70% concentrate, Diet I and II were supplemented with a SC. Daily urine was collected and stored at -20°C until analysis. Calorimetric methods were used for the determination of urinary allantoin and creatinin levels. The estimated microbial N supply to small intestine for Diets I, I+SC, II and II+SC were 2.51, 2.64, 2.95 and 3.43 g N/d respectively. Supplementation of Diets I and II with SC significantly affected the allantoin levels in µmol/W0. 75 (p<0.05). Mean creatinine values in µmol/W0. 75 and allantoin:creatinin ratios were not significantly different among diets. In conclusion, supplementation with SC live yeast culture had a significant effect on urinary allantoin excretion and microbial protein supply to small intestine in Kivircik yearlings fed with high concentrate Diet II (P<0.05). Hence urinary allantoin excretion may be used as a tool for estimating microbial protein supply in Kivircık yearlings. However, further studies are necessary to understand the metabolism of Saccharomyces cerevisiae live yeast culture with different forage: concentrate ratio in Kıvırcık Yearlings.

Keywords: allantoin, creatinin, Kivircik yearling, microbial nitrogen, Saccharomyces cerevisia

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Authors: Sonaal Ramsookmohan

Abstract:

Legume seeds are common foods in human diet and have been identied as a valuable source of human nutritonn Since they are useful sources of protein; legume proteins are used in many food applicatonsn Critcal functonal propertes are recognized to impact the quality of foodn Cowpea (Vigna unguiculata), has been well documented for its immense potental in contributng to food security forming part of daily staple diets in most developing countriesn. In this study, cowpea seeds were used to prepare cowpea four, protein isolates by the salt extractonndialysis method and peptdes by enzymatc hydrolysis using Alcalase and Flavourzymen Functonal analyses such as water absorpton capacity, oil absorpton capacity, emulsifying and foaming propertes were conducted on the cowpea peptdesn The physicochemical propertes determine their potental applicaton in food industries as functonal ingredientsn Cowpea peptdes could increase the value of cowpea by expanding its use, as well as contribute to the legume grain sector.

Keywords: physicochemical, peptides, Cowpea, alcalase, flavourzyme

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Authors: Daniel Assefa, Engeda Dessalegn, Chetan Chauhan

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Moringa stenopetala is a socioeconomic valued tree that is widely available and cultivated in the Southern part of Ethiopia. The leaves have been traditionally used as a food source with high nutritional and medicinal values. The present work was carried out to evaluate the effect of thermal treatment on the total phenolic content, antioxidant and alpha-amylase inhibition activities of aqueous leaf extracts during maceration and different decoction time interval (5, 10 and 15 min). The total phenolic content was determined by the Folin-ciocalteu methods whereas antioxidant activities were determined by 2,2-diphenyl-1-picryl-hydrazyl(DPPH) radical scavenging, reducing power and ferrous ion chelating assays and alpha-amylase inhibition activity was determined using 3,5-dinitrosalicylic acid method. Total phenolic content ranged from 34.35 to 39.47 mgGAE/g. Decoction for 10 min extract showed ferrous ion chelating (92.52), DPPH radical scavenging (91.52%), alpha-amylase inhibition (69.06%) and ferric reducing power (0.765), respectively. DPPH, reducing power and alpha-amylase inhibition activities showed positive linear correlation (R2=0.853, R2= 0.857 and R2=0.930), respectively with total phenolic content but ferrous ion chelating activity was found to be weakly correlated (R2=0.481). Based on the present investigation, it could be concluded that major loss of total phenolic content, antioxidant and alpha-amylase inhibition activities of the crude leaf extracts of Moringa stenopetala leaves were observed at decoction time for 15 min. Therefore, to maintain the total phenolic content, antioxidant, and alpha-amylase inhibition activities of leaves, cooking practice should be at the optimum decoction time (5-10 min).

Keywords: alpha-amylase inhibition, antioxidant, Moringa stenopetala, total phenolic content

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Authors: Rezvan Najafi, Korosh Heydari, Massoud Saidijam

Abstract:

5-FU is a chemotherapeutic agent that has been used in colorectal cancer (CRC) treatment. However, it is usually associated with the acquired resistance, which decreases the therapeutic effects of 5-FU. miR-200c is involved in chemotherapeutic drug resistance, but its mechanism is not fully understood. In this study, the effect of inhibition of miR-200c in sensitivity of HCT-116 CRC cells to 5-FU was evaluated. HCT-116 cells were transfected with LNA-anti- miR-200c for 48 h. mRNA expression of miR-200c was evaluated using quantitative real- time PCR. The protein expression of phosphatase and tensin homolog (PTEN) and E-cadherin were analyzed by western blotting. Annexin V and propidium iodide staining assay were applied for apoptosis detection. The caspase-3 activation was evaluated by an enzymatic assay. The results showed LNA-anti-miR-200c inhibited the expression of PTEN and E-cadherin protein, apoptosis and activation of caspase 3 compared with control cells. In conclusion, these results suggest that miR-200c as a prognostic marker can overcome to 5-FU chemoresistance in CRC.

Keywords: colorectal cancer, miR-200c, 5-FU resistance, E-cadherin, PTEN

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Authors: Gonzalo Ortiz de Elguea-Culebras, Raúl Sánchez-Vioquea, Adela Mena-Morales, Manuel Alaiz, Enrique Melero-Bravo, Esteban García-Romero, Javier Vioque, Lourdes Marchante-Cuevas, Julio Girón-Calle

Abstract:

Saffron (Crocus sativus L.) is a highly valued crop for the manufacture of spice that consists of the dried stigma of the flowers. This is in contrast to other underutilized parts of the saffron plant as leaves, which represent abundant biomass whose use might help to enhance the sustainability of the saffron crop. Saffron leaves contain significant amounts of phenolic compounds, 7.8 equivalent grams of gallic acid per 100g of extract, and are very promising compounds in terms of exploring novel uses of saffron leaves. Given that phenolic compounds have numerous effects on cancer-related biological pathways, we have investigated the in vitro antiproliferative effect of saffron leaf polyphenols against human colon adenocarcinoma cells (Caco-2). Polyphenols were extracted from leaves with 70% ethanol, defatted with hexane, and purified by solid phase extraction using C18 silica gel and then silica gel 60. Analysis of polyphenols was performed by HPLC-ESI-MS. Di-, tri-, and tetrahexosides of quercetin, kaempferol, and isorhamnetin, as well as C-hexosides like isoorientin and vitexin, were tentatively identified. Polyphenols strongly inhibited the proliferation of Caco-2 cells, which is consistent with model studies in which several of the polyphenols identified in saffron leaves have demonstrated their potential as chemopreventive agents in cancer. Due to the low profitability that saffron leaf currently represents, we consider these results very encouraging and that this by-product deserves further investigation as a potential source of active molecules against colorectal cancer.

Keywords: saffron leaves, agricultural by-products, polyphenols, antiproliferative effect, human colon adenocarcinoma cells

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