Search results for: biological reactor

Authors: Sharmi Mukherjee, Anindita Chakraborty

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Cellular irradiation incites complex responses including arrest of cell cycle progression. This article accentuates the effects of radiation on cell cycle status of radiation non-targeted cells. Human Hepatoma HepG2 cells were exposed to increasing doses of γ radiations (1, 2, 4, 6 Gy) and their cell culture media was transferred to non-targeted HepG2 cells cultured in other Petri plates. These radiation non-targeted cells cultured in the ICCM (Irradiated cell conditioned media) were the bystander cells on which cell cycle analysis was performed using flow cytometry. An apparent decrease in the distribution of bystander cells at G0/G1 phase was observed with increased radiation doses upto 4 Gy representing a linear relationship. This was accompanied by a gradual increase in cellular distribution at G2/M phase. Interestingly the number of cells in G2/M phase at 1 and 2 Gy irradiation was not significantly different from each other. However, the percentage of G2 phase cells at 4 and 6 Gy doses were significantly higher than 2 Gy dose indicating the IC50 dose to be between 2 and 4 Gy. Cell cycle arrest is an indirect indicator of genotoxic damage in cells. In this study, bystander stress signals through the cell culture media of irradiated cells disseminated the radiation induced DNA damages in the non-targeted cells which resulted in arrest of the cell cycle progression at G2/M phase checkpoint. This implies that actual radiation biological effects represent a penumbra with effects encompassing a larger area than the actual beam. This article highlights the existence of genotoxic damages as bystander effects of γ rays in human Hepatoma cells by cell cycle analysis and opens up avenues for appraisal of bystander stress communications between tumor cells. Contemplation of underlying signaling mechanisms can be manipulated to maximize damaging effects of radiation with minimum dose and thus has therapeutic applications.

Keywords: bystander effect, cell cycle, genotoxic damage, hepatoma

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Authors: Abdullah Al-Nassri, Ahmed Al-Alawi

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Seaweeds are classified into three groups: red, green, and brown. Each group of seaweeds consists of several types that have differences in composition. Even at the species level, there are differences in some ingredients, although in general composition, they are the same. Environmental conditions, availability of nutrients, and maturity stage are the main reasons for composition differences. In this study, two red seaweed species, Melanothamnus somalensis & Gelidium omanense, were collected in September 2021 from Sadh (Dhofar governorate, Oman). Five organic solvents were used sequentially to achieve extraction. The solvents were applied in the following order: hexane, dichloromethane, ethyl acetate, acetone, and methanol. Preparative HPLC (PrepLC) was performed to fraction the extracts. The chemical composition was measured; also, total phenols, flavonoids, and tannins were investigated. The structure of the extracts was analyzed by Fourier-transform infrared spectroscopy (FTIR). Seaweeds demonstrated high differences in terms of chemical composition, total phenolic content (TPC), total flavonoid content (TFC), and total tannin content (TTC). Gelidium omanense showed high moisture content, lipid content and carbohydrates (9.8 ± 0.15 %, 2.29 ± 0.09 % and 70.15 ± 0.42 %, respectively) compared to Melanothamnus somalensis (6.85 ± 0.01 %, 2.05 ± 0.12 % and 52.7 ± 0.36 % respectively). However, Melanothamnus somalensis showed high ash content and protein (27.68 ± 0.40 % and 52.7 ± 0.36 % respectively) compared to Gelidium omanense (8.07 ± 0.39 % and 9.70 ± 0.22 % respectively). Melanothamnus somalensis showed higher elements and minerals content, especially sodium and potassium. This is attributed to the jelly-like structure of Melanothamnus somalensis, which allows storage of more solutes compared to the leafy-like structure of Gelidium omanense. Furthermore, Melanothamnus somalensis had higher TPC in all fractions except the hexane fraction than Gelidium omanense. Except with hexane, TFC in the other solvents’ extracts was significantly different between Gelidium omanense and Melanothamnus somalensis. In all fractions, except dichloromethane and ethyl acetate fractions, there were no significant differences in TTC between Gelidium omanense and Melanothamnus somalensis. FTIR spectra showed variation between fractions, which is an indication of different functional groups.

Keywords: chemical composition, organic extract, Omani seaweeds, biological activity, FTIR

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Authors: Saule Shazhanbayeva, Denise van der Merwe

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Study hypothesis: Nazarbayev Intellectual School of Kyzylorda’s Biology teachers can use STEM-integrated learning to improve students' problem-solving ability and responsibility as global citizens. The significance of this study is to indicate how the use of STEM integrative learning during Biology lessons could contribute to forming globally-minded students who are responsible community members. For the purposes of this study, worldview is defined as a view that is broader than the country of Kazakhstan, allowing students to see the significance of their scientific contributions to the world as global citizens. The context of worldview specifically indicates that most students have never traveled outside of their city or region within Kazakhstan. In order to broaden student understanding, it is imperative that students are exposed to different world views and contrasting ideas within the educational setting of Biology as the science being used for the research. This exposure promulgates students understanding of the significance they have as global citizens alongside the obligations which would rest on them as scientifically minded global citizens. Integrative learning should be Biological Science - with Technology and engineering in the form of problem-solving, and Mathematics to allow improved problem-solving skills to develop within the students of Nazarbayev Intellectual School (NIS) of Kyzylorda. The school's vision is to allow students to realise their role as global citizens and become responsible community members. STEM allows integrations by combining four subject skills to solve topical problems designed by educators. The methods used are based on qualitative analysis: for students’ performance during a problem-solution scenario; and Biology teacher interviews to ascertain their understanding of STEM implementation and willingness to integrate it into current lessons. The research indicated that NIS is ready for a shift into STEM lessons to promote globally responsible students. The only additional need is for proper STEM integrative lesson method training for teachers.

Keywords: global citizen, STEM, Biology, high-school

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Authors: Vidia A. Nuraini, Eugene M. H. Yee, Mohan Bhadbhade, David StC. Black, Naresh Kumar

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Flavonoids are naturally occurring compounds that have been shown to exhibit a wide range of biological properties including anticancer and anti-inflammatory activities. However, flavonoids suffer from low bioavailability, which limits their overall utility for therapeutic applications. One of the methods to overcome this limitation is through structural modification of natural flavonoids. In this study, flavanone, isoflavanone, and isoflavene, were structurally modified through the introduction of additional fused-ring systems via ortho-quinone methide intermediates (o-QMs). These intermediates can readily undergo a [4+2] cycloaddition through an inverse-electron-demand Diels–Alder reaction with electron-rich dienophiles. A regioselective Mannich reaction using bis-(N,N-dimethylamino)methane was employed to generate the o-QM precursors of flavanone, isoflavanone, and isoflavene. The o-QM intermediates were subsequently generated in situ through thermal elimination of the dimethylamine functionality and reacted with a variety of dienophiles to produce novel flavonoids with fused-ring systems. A total of 21 novel flavonoid analogs were successfully synthesized. The X-ray crystal structure of cycloaddition adducts, particularly those derived from 3,4-dihydro-2H-pyran and p-methoxystyrene revealed a special case of enantiomeric disorder, where two enantiomers in equal amounts superpose with one another, with the exception for atoms that have opposite configuration. The anticancer properties of fused-ring systems derived from isoflavene were evaluated against the neuroblastoma SKN-BE(2)C, the triple negative breast cancer MDA-MB-231, and the glioblastoma U87 cancer cell lines. One of these cycloaddition adducts had displayed improved anti-proliferative activity against MDA-MB-231 and U87 cancer cell lines as compared to the parent compound. Further anticancer and anti-inflammatory activities of the flavanone and isoflavanone analogs are currently being investigated.

Keywords: Diels-Alder reaction, flavonoids, Mannich reaction, ortho-quinone methide.

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Authors: Mouna Baassi, Mohamed Moussaoui, Sanchaita Rajkhowa, Hatim Soufi, Said Belaaouad

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The objective of this paper is to address the challenging task of targeting Human Immunodeficiency Virus type 1 Reverse Transcriptase (HIV-1 RT) in the treatment of AIDS. Reverse Transcriptase inhibitors (RTIs) have limitations due to the development of Reverse Transcriptase mutations that lead to treatment resistance. In this study, a combination of statistical analysis and bioinformatics tools was adopted to develop a mathematical model that relates the structure of compounds to their inhibitory activities against HIV-1 Reverse Transcriptase. Our approach was based on a series of compounds recognized for their HIV-1 RT enzymatic inhibitory activities. These compounds were designed via software, with their descriptors computed using multiple tools. The most statistically promising model was chosen, and its domain of application was ascertained. Furthermore, compounds exhibiting comparable biological activity to existing drugs were identified as potential inhibitors of HIV-1 RT. The compounds underwent evaluation based on their chemical absorption, distribution, metabolism, excretion, toxicity properties, and adherence to Lipinski's rule. Molecular docking techniques were employed to examine the interaction between the Reverse Transcriptase (Wild Type and Mutant Type) and the ligands, including a known drug available in the market. Molecular dynamics simulations were also conducted to assess the stability of the RT-ligand complexes. Our results reveal some of the new compounds as promising candidates for effectively inhibiting HIV-1 Reverse Transcriptase, matching the potency of the established drug. This necessitates further experimental validation. This study, beyond its immediate results, provides a methodological foundation for future endeavors aiming to discover and design new inhibitors targeting HIV-1 Reverse Transcriptase.

Keywords: QSAR, ADMET properties, molecular docking, molecular dynamics simulation, reverse transcriptase inhibitors, HIV type 1

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Authors: Adamu Muhammed Tukur

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Pulp and paper industry is one of the fastest growing industries due to an increased demand in paper products. For it to satisfy this ever increasing demand, it adopts new technological innovations some of which are proved to affect our environment negatively. Global consumption of paper has increased by 400% in the last four decades and this suggests that more research is required to assess the impact of industrial effluents to our environment and public health. Paper products are generally biodegradable, however, the processes involved in its production which involve the use of mainly bleaching agents and other non-biodegradable substances pose serious problem to the environment. There are more than 250 chemicals released in paper mill waste and some are xenobiotics. Different methods such as physical and chemical methods can be adopted for the remediation of the effluents but are proved to be costly and not safe to the environment. On the other hand, biological method is shown to be less costly and environmentally friendly. Microorganisms and their enzymes have shown a promising future for bioremediation of effluents related to paper mill. Many studies prove that one of the major pollutants in the paper mill effluent is phenol especially its chlorinated derivatives. Pentachlorophenol is extremely hazardous to living cells and therefore need to be removed from the environment. Microorganisms including bacteria and fungi have the potential to degrade phenolic compounds e.g. Bacillus stearothermiphilus, Pseudomonas putida, Coricus versicolor, Sphingomonas chlorophenolica, Fusarium sp, Bacillus subtilis and P. aeroginosa. Enzymes used for the degradation include phenol hydrooxylase, polyphenoloxylase, laccase, peroxidase among others. Lignin is another important pollutant and is resistant to microbial degradation but it has been proved that certain bacteria and fungi like can degrade it. Among the fungi white-rot fungi like Fomes lividus and Trametes vesicolor are the most important bioremediators. This review focused on use of microorganism to reduce or eradicate pollutants released from the paper industry. It can serve as a review for further research to be conducted especially in the field of Biotechnology.

Keywords: bioremediation, pulp and paper, pentachlorophenol, environment

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Authors: Danielle Katrina Santos, Koji Shimada

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Environmentally-friendly agriculture has been promoted for over two decades as a response to the environmental challenges brought by climate change and biological loss. Located above the equator, it is possible that Japan may benefit from future climate change, yet Japan is also a rarely developed country located in the Asian Monsoon climate region, making it vulnerable to the impacts of climate change. In this regard, the Japanese government has initiated policies to adapt to the adverse effects of climate change through the promotion and popularization of environmentally-friendly farming practices. This study aims to determine profit efficiency among environmentally-friendly rice farmers in Shiga Prefecture using the Stochastic Frontier Approach. A cross-sectional survey was conducted among 66 farmers from top rice-producing cities through a structured questionnaire. Results showed that the gross farm income of environmentally-friendly rice farmers was higher by JPY 316,223.00/ha. Production costs were also found to be higher among environmentally-friendly rice farmers, especially on labor costs, which accounted for 32% of the total rice production cost. The resulting net farm income of environmentally-friendly rice farmers was only higher by JPY 18,044/ha. Results from the stochastic frontier analysis further showed that the profit efficiency of conventional farmers was only 69% as compared to environmentally-friendly rice farmers who had a profit efficiency of 76%. Furthermore, environmentally-friendly agriculture participation, other types of subsidy, educational level, and farm size were significant factors positively influencing profit efficiency. The study concluded that substitution of environmentally-friendly agriculture for conventional rice farming would result in an increased profit efficiency due to the direct payment subsidy and price premium received. The direct government policies that would strengthen the popularization of environmentally-friendly agriculture to increase the production of environmentally-friendly products and reduce pollution load to the Lake Biwa ecosystem.

Keywords: profit efficiency, environmentally-friendly agriculture, rice farmers, direct payment subsidies

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Authors: Joanna Skopinska-Wisniewska, Anna Bajek, Marta Ziegler-Borowska, Alina Sionkowska

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Hydrogels are a class of materials widely used in medicine for many years. Proteins, such as collagen, due to the presence of a large number of functional groups are easily wettable by polar solvents and can create hydrogels. The supramolecular network capable to swelling is created by cross-linking of the biopolymers using various reagents. Many cross-linking agents has been tested for last years, however, researchers still are looking for a new, more secure reactants. Squaric acid, 3,4-dihydroxy 3-cyclobutene 1,2- dione, is a very strong acid, which possess flat and rigid structure. Due to the presence of two carboxyl groups the squaric acid willingly reacts with amino groups of collagen. The main purpose of this study was to investigate the influence of addition of squaric acid on the chemical, physical and biological properties of collagen materials. The collagen type I was extracted from rat tail tendons and 1% solution in 0.1M acetic acid was prepared. The samples were cross-linked by the addition of 5%, 10% and 20% of squaric acid. The mixtures of all reagents were incubated 30 min on magnetic stirrer and then dialyzed against deionized water. The FTIR spectra show that the collagen structure is not changed by cross-linking by squaric acid. Although the mechanical properties of the collagen material deteriorate, the temperature of thermal denaturation of collagen increases after cross-linking, what indicates that the protein network was created. The lyophilized collagen gels exhibit porous structure and the pore size decreases with the higher addition of squaric acid. Also the swelling ability is lower after the cross-linking. The in vitro study demonstrates that the materials are attractive for 3T3 cells. The addition of squaric acid causes formation of cross-ling bonds in the collagen materials and the transparent, stiff hydrogels are obtained. The changes of physicochemical properties of the material are typical for cross-linking process, except mechanical properties – it requires further experiments. However, the results let us to conclude that squaric acid is a suitable cross-linker for protein materials for medicine and tissue engineering.

Keywords: collagen, squaric acid, cross-linking, hydrogel

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Authors: Kamila Weissova, Jitka Skrabalova, Katerina Skalova, Jana Koprivova, Zdenka Bendova

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Any Arctic visitor has to deal with extreme conditions, including constant light during the summer season or constant darkness during winter time. Light/dark cycle is the most powerful synchronizing signal for biological clock and the absence of daily dark period during the polar day can significantly alter the functional state of the internal clock. However, the inner clock can be synchronized by other zeitgebers such as physical activity, food intake or social interactions. Here, we investigated the effect of polar day on circadian clock of 10 researchers attending the polar base station in the Svalbard region during July. The data obtained on Svalbard were compared with the data obtained before the researchers left for the expedition (in the Czech Republic). To determine the state of circadian clock we used wrist actigraphy followed by sleep diaries, saliva, and buccal mucosa samples, both collected every 4 hours during 24h-interval to detect melatonin by radioimmunoassay and clock gene (PER1, BMAL1, NR1D1, DBP) mRNA levels by RT-qPCR. The clock gene expression was analyzed using cosinor analysis. From our results, it is apparent that the constant sunlight delayed melatonin onset and postponed the physical activity in the same order. Nevertheless, the clock gene expression displayed higher amplitude on Svalbard compared to the amplitude detected in the Czech Republic. These results have suggested that the common daily schedule at the Svalbard expedition can strengthen circadian rhythm in the environment that is lacking light/dark cycle. In conclusion, the constant sunlight delays melatonin onset, but it still maintains its rhythmic secretion. The effect of constant sunlight on circadian clock can be minimalized by common daily scheduled activity.

Keywords: actighraph, clock genes, human, melatonin, polar day

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Authors: P. Susheela, Mary Rosaline, R. Radha

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This research was designed to determine the bioactive compounds present in the nest samples of the mud dauber wasp, Sceliophron caementarium. Insects and insect-based products have been used for the treatment of various ailments from a very long time. It has been found that all over the world including the western societies and the indigenous populations, the usage of insect-based medicine plays an important role in various healing practices and magic rituals. Studies on the therapeutic usage of insects are negligible when compared to plants, the. In the present scenario, it is important to explore bioactive compounds from natural sources rather than depending on synthetic drugs that have adverse effects on human body. Keeping this in view, an attempt was made to analyze and identify bioactive components from the nest sample of the mud dauber wasp, Sceliophron caementarium. The nests of the mud dauber wasp, Sceliophron caementarium were collected from Coimbatore, Tamil Nadu, India. The nest sample was extracted with ethanol for 6-8 hours using Soxhlet apparatus. The final residue was obtained by filtering the extract through Whatman filter paper No.41. The GCMS analysis of the nest sample was performed using Perkin Elmer Elite - 5 capillary column. The resultant compounds were compared with the database of National Institute Standard and Technology (NIST), WILEY8, FAME. The GC-MS analysis of the concentrated ethanol extract revealed the presence of eight constituents like Methylene chloride, Eicosanoic acid, 1, 1’:3’, 1’’-Terphenyl, 5'-Phenyl, Di-N-Decylsulfone, 1, 2-Bis (Trimethylsilyl) Benzene, Androstane-11, 17-Dione, 3-[(Trimethylsilyl) Oxy]-, 17-[O-(Phenylmethyl) O. Most of the identified compounds were reported as having biological activities viz. anti-inflammatory, antibacterial and antifungal properties that can be of pharmaceutical importance and further study of these isolated compounds may prove their medicinal importance in future.

Keywords: Sceliophron caementarium, Gas chromatography-mass spectrometry, ethanol extract, bioactive compounds

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Authors: Parvaneh Mahmoudi, Ahmad Moeni, Seyed Mojtaba Khayam Nekoei, Mohsen Mardi, Mehrshad Zeinolabedini, Ghasem Hosseini Salekdeh

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Saffron (Crocus sativus L.) is one of the most important spice and medicinal plants. The three-branch style of C. sativus flowers are the most important economic part of the plant and known as saffron, which has several medicinal properties. Despite the economic and biological significance of this plant, knowledge about its molecular characteristics is very limited. In the present study, we, for the first time, constructed a comprehensive dataset for C. sativus stigma through de novo transcriptome sequencing. We performed de novo transcriptome sequencing of C. sativus stigma using the Illumina paired-end sequencing technology. A total of 52075128 reads were generated and assembled into 118075 unigenes, with an average length of 629 bp and an N50 of 951 bp. A total of 66171unigenes were identified, among them, 66171 (56%) were annotated in the non-redundant National Center for Biotechnology Information (NCBI) database, 30938 (26%) were annotated in the Swiss-Prot database, 10273 (8.7%) unigenes were mapped to 141 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database, while 52560 (44%) and 40756 (34%) unigenes were assigned to Gen Ontology (GO) categories and Eukaryotic Orthologous Groups of proteins (KOG), respectively. In addition, 65 candidate genes involved in three stages of crocin biosynthesis were identified. Finally, transcriptome sequencing of saffron stigma was used to identify 6779 potential microsatellites (SSRs) molecular markers. High-throughput de novo transcriptome sequencing provided a valuable resource of transcript sequences of C. sativus in public databases. In addition, most of candidate genes potentially involved in crocin biosynthesis were identified which could be further utilized in functional genomics studies. Furthermore, numerous obtained SSRs might contribute to address open questions about the origin of this amphiploid spices with probable little genetic diversity.

Keywords: saffron, transcriptome, NGS, bioinformatic

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Authors: Tariq Ahmad Dar, Moinuddin, M. Masroor A. Khan

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There is considerable need in enhancing the content and yield of active constituents of medicinal plants keeping in view their massive demand worldwide. Different strategies have been employed to enhance the active constituents of medicinal plants and the use of phytohormones has been proved effective in this regard. Gamma-irradiated Sodium alginate (ISA) is known to elicit an array of plant defense responses and biological activities in plants. Considering the medicinal importance, a pot experiment was conducted to explore the effect of ISA and phosphorus on growth, yield and quality of fenugreek (Trigonella foenum-graecum L.). ISA spray treatments (0, 40, 80 and 120 mg L-1) were applied alone and in combination with 40 kg P ha-1 (P40). Crop performance was assessed in terms of plant growth characteristics, physiological attributes, seed yield and the content of seed trigonelline. Of the ten-treatments, P40 + 80 mg L−1 of ISA proved the best. The results showed that foliar spray of ISA alone or in combination with P40 augmented the plant vegetative growth, enzymatic activities, trigonelline content, trigonelline yield and economic yield of fenugreek. Application of 80 mg L−1 of ISA applied with P40 gave the best results for almost all the parameters studied compared to control or to 80 mg L−1 of ISA applied alone. This treatment increased the total content of chlorophyll, carotenoids, leaf -N, -P and -K and trigonelline compared to the control by 24.85 and 27.40%, 15 and 23.52%, 18.70 and 16.84%, 15.88 and 18.92%, 12 and 14.44%, at 60 and 90 DAS respectively. The combined application of 80 mg L−1 of ISA along with P40 resulted in the maximum increase in seed yield, trigonelline content and trigonelline yield by146, 34 and 232.41%, respectively, over the control. Gel permeation chromatography revealed the formation of low molecular weight fractions in ISA samples, containing even less than 20,000 molecular weight oligomers, which might be responsible for plant growth promotion in this study. Trigonelline content was determined by reverse phase high performance liquid chromatography (HPLC) with C-18 column.

Keywords: gamma-irradiated sodium alginate, phosphorus, gel permeation chromatography, HPLC, trigonelline content, yield

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Authors: Renata Gagić-Serdar, Miroslava Markovic, Ljubinko Rakonjac, Aleksandar Lučić

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The paper present the results of the entomological experimental studies of the biological, ecological, and (bionomic) insect performances, such as seasonal adaptation of introduced monophagous false indigo and albizias weevil’s Acanthoscelides pallidipennis Motschulsky. and Bruchidius terrenus (Sharp), Coleoptera: Chrysomelidae: Bruchinae, to phenological phases of aggressive invasive host plant Amorpha fruticosa L. and Albizia julibrissin (Fabales: Fabaceae) on the territory of Republic of Serbia with special attention on assessing and monitoring of new formed and detected inter species relations between autochthons parasite wasps from fauna (Hymenoptera: Chalcidoidea) and herbaceous seed weevil beetle. During 15 years (2006-2021), on approximately 30 localities, data analyses were done for observed experimental host plants from samples with statistical significance. Status of genera from families Hymenoptera: Chalcidoidea.: Pteromalidae and Eulophidae, after intensive investigations, has been trophicly identified. Recorded seed pest species of A. fruticosa or A. julibrissin (Fabales: Fabaceae) was introduced in Serbia and planted as ornamental trees, they also were put undergo different kinds of laboratory and field research tests during this period in a goal of collecting data about lasting each of develop stage of their seed beetles. Field generations in different stages were also monitored by continuous infested seed collecting and its disection. Established host plant-seed predator linkage was observed in correlation with different environment parameters, especially water level fluctuations in bank corridor formation stands and riparian cultures.

Keywords: amorpha, albizia, chalcidoid wasp, invasiveness, weevils

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Authors: E.Tskitisvili, L. Jgenti, I. Eliava, T. Tskitishvili, N. Bagathuria, M. Gigolashvili

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The fight against dangerous nematode diseases that have world economic importance requires accurate data about the prevalence of these pests. In the point of view of the International Convention on Biological Diversity, the identification of the plant invasion causing dangerous pathogen in the early stages of invasion on new territory is the most important part of the program, which aims to monitor the Bio-Agro Coenosis and Bio-Control. Citrus nematode-specific belongs to the pathogen species, which can cause epiphytotics particularly for large areas and cause irreparable damage to citrus plantations. This paper provides a brief tour of the spread of citrus nematodes on the Black Sea coast (Adjara and Abkhazia). Also the bio-ecological monitoring data to detect the potential sources of invasion for evaluating the current conditions of the citrus nematodes prevalence. Through 2006-2010, the material was gained by structural monitoring system during the citrus vegetation period on tangerines, lemon and oranges from nine points of the study area. Mature forms of Tylenchulus semipenetrans Cobb, 1913 were observed in almost all of the samples of the root system, the peak of larvae was observed in late spring and outumn. 92 forms of nematode has been detected in the rhizosphere belonging to 8 Orders: Areolaimida, Dorylaimida, Enoplida, Mononchida, Tylenshida, Monshysterida, Rhabditida, Aphelenchida, 23 families and 40 genera. 75 forms are identified as species. It is estimated the number of nematodes fauna and ecological groups. To detect possible sources of invasion we obtained additional materials in 2013-2014 from citrus plantations planted in 2011, where is planted tangerine trees introduced from Spain and Japan. The fauna of rhizosphere is identified and Tylenchulus semipenetrans Cobb, 1913 is not detected.

Keywords: Citrus nematodes, infection, bioecological monitoring, epiphytotics

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Authors: Mudrika Singhal

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The research paper is basically about how the genetically modified organisms evolved and their significance in today’s world. It also highlights about the various pros and cons of the genetically modified organisms and the progress of India in this field. A genetically modified organism is the one whose genetic material has been altered using genetic engineering techniques. They have a wide range of uses such as transgenic plants, genetically modified mammals such as mouse and also in insects and aquatic life. Their use is rooted back to the time around 12,000 B.C. when humans domesticated plants and animals. At that humans used genetically modified organisms produced by the procedure of selective breeding and not by genetic engineering techniques. Selective breeding is the procedure in which selective traits are bred in plants and animals and then are domesticated. Domestication of wild plants into a suitable cultigen is a well known example of this technique. GMOs have uses in varied fields ranging from biological and medical research, production of pharmaceutical drugs to agricultural fields. The first organisms to be genetically modified were the microbes because of their simpler genetics. At present the genetically modified protein insulin is used to treat diabetes. In the case of plants transgenic plants, genetically modified crops and cisgenic plants are the examples of genetic modification. In the case of mammals, transgenic animals such as mice, rats etc. serve various purposes such as researching human diseases, improvement in animal health etc. Now coming upon the pros and cons related to the genetically modified organisms, pros include crops with higher yield, less growth time and more predictable in comparison to traditional breeding. Cons include that they are dangerous to mammals such as rats, these products contain protein which would trigger allergic reactions. In India presently, group of GMOs include GM microorganisms, transgenic crops and animals. There are varied applications in the field of healthcare and agriculture. In the nutshell, the research paper is about the progress in the field of genetic modification, taking along the effects in today’s world.

Keywords: applications, mammals, transgenic, engineering and technology

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Authors: Pelin Yılmaz Çetiner, Metin Mert İlgün, Nazlı Çetindağ, Emine Birci, Gizemnur Yıldız Uysal, Özcan Hatipoğlu, Ehsan Tuzcuoğlu, Gökhan Sır

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Water scarcity is an inevitable problem affecting more and more people day by day. It is a worldwide crisis and a consequence of rapid population growth, urbanization and overexploitation. Thus, the solutions providing the reclamation of the wastewater are the desired approach. Wastewater contains various substances such as organic, soaps and detergents, solvents, biological substances, and inorganic substances. The physical properties of the wastewater differs regarding to its origin such as commerical, domestic or hospital usage. Thus, the treatment strategy of this type of wastewater is should be comprehensively investigated and properly treated. The advanced oxidation process comes up as a hopeful method associated with the formation of reactive hydroxyl radicals that are highly reactive to oxidize of organic pollutants. This process has a priority on other methods such as coagulation, flocuation, sedimentation and filtration since it was not cause any undesirable by-products. In the present study, it was aimed to investigate the applicability of advanced oxidation process for the treatment of household appliances wastewater. For this purpose, the laboratory studies providing the effectively addressing of the formed radicals to organic pollutants were carried out. Then the effect of process parameters were comprehensively studied by using response surface methodology, Box-Benhken experimental desing. The final chemical oxygen demand (COD) was the main output to evaluate the optimum point providing the expected COD removal. The linear alkyl benzene sulfonate (LAS), total dissolved solids (TDS) and color were measured for the optimum point providing the expected COD removal. Finally, present study pointed out that advanced oxidation process might be efficiently preffered to treat of the household appliances wastewater and the optimum process parameters provided that expected removal of COD.

Keywords: advanced oxidation process, household appliances wastewater, modelling, water reuse

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Authors: Hui-Chen Tsai, Chien-Jen Ho, Bo-Wei Power Liang, Ying Shen, Yi-Hsin Lai

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Former Taiwan Metal Mining Corporation and its associated 3 wasted flue gas tunnels, hereinafter referred to as 'TMMC', was contaminated with heavy metals, Polychlorinated biphenyls (PCBs) and Total Petroleum Hydrocarbons (TPHs) in soil. Since the contamination had been exposed and unmanaged in the environment for more than 40 years, the extent of the contamination area is estimated to be more than 25 acres. Additionally, TMMC is located in a remote, mountainous area where almost no residents are residing in the 1-km radius area. Thus, it was deemed necessary to conduct an ecological risk assessment in order to evaluate the details of future contaminated site management plan. According to the winter and summer, ecological investigation results, one type of endangered, multiple vulnerable and near threaten plant was discovered, as well as numerous other protected species, such as Crested Serpent Eagle, Crested Goshawk, Black Kite, Brown Shrike, Taiwan Blue Magpie were observed. Ecological soil screening level (Eco-SSLs) developed by USEPA was adopted as a reference to conduct screening assessment. Since all the protected species observed surrounding TMMC site were birds, screening ecological risk assessment was conducted on birds only. The assessment was assessed mainly based on the chemical evaluation, which the contamination in different environmental media was compared directly with the ecological impact levels (EIL) of each evaluation endpoints and the respective hazard quotient (HQ) and hazard index (HI) could be obtained. The preliminary ecological risk assessment results indicated HI is greater than 1. In other words, the biological stressors (birds) were exposed to the contamination, which was already exceeded the dosage that could cause unacceptable impacts to the ecological system. This result was mainly due to the high concentration of arsenic, metal and lead; thus it was suggested the above mention contaminants should be remediated as soon as possible or proper risk management measures should be taken.

Keywords: screening, ecological risk assessment, ecological impact levels, risk management

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Authors: Elisa K. Schoenell, Cristiano De Oliveira, Luiz R. H. Dos Santos, Alexandre Giacobbo, Andréa M. Bernardes, Marco A. S. Rodrigues

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Water is an indispensable natural resource, which must be preserved to human activities as well the ecosystems. However, the sewage discharge has been contaminating water resources. Conventional treatment, such as physicochemical treatment followed by biological processes, has not been efficient to the complete degradation of persistent organic compounds, such as medicines and hormones. Therefore, the use of advanced technologies to sewage treatment has become urgent and necessary. The aim of this study was to apply Reverse Osmosis (RO) on sewage tertiary treatment from a Waste Water Treatment Plant (WWTP) in south Brazil. It was collected 200 L of sewage pre-treated by wetland with aquatic macrophytes. The sewage was treated in a RO pilot plant, using a polyamide membrane BW30-4040 model (DOW FILMTEC), with 7.2 m² membrane area. In order to avoid damage to the equipment, this system contains a pleated polyester filter with 5 µm pore size. It was applied 8 bar until achieve 5 times of concentration, obtaining 80% of recovery of permeate, with 10 L.min-1 of concentrate flow rate. Samples of sewage pre-treated on WWTP, permeate and concentrate generated on RO was analyzed for physicochemical parameters and by gas chromatography (GC) to qualitative analysis of organic compounds. The results proved that the sewage treated on WWTP does not comply with the limit of phosphorus and nitrogen of Brazilian legislation. Besides this, it was found many organic compounds in this sewage, such as benzene, which is carcinogenic. Analyzing permeate results, it was verified that the RO as sewage tertiary treatment was efficient to remove of physicochemical parameters, achieving 100% of iron, copper, zinc and phosphorus removal, 98% of color removal, 91% of BOD and 62% of ammoniacal nitrogen. RO was capable of removing organic compounds, however, it was verified the presence of some organic compounds on de RO permeate, showing that RO did not have the capacity of removal all organic compounds of sewage. It has to be considered that permeate showed lower intensity of peaks in chromatogram in comparison to the sewage of WWTP. It is important to note that the concentrate generate on RO needs a treatment before its disposal in environment.

Keywords: organic compounds, reverse osmosis, sewage treatment, tertiary treatment

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Authors: Neelma Ashraf

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Actinomycetes, particularly genus Streptomyces is of great importance due to their role in the discovery of new natural products, particularly antimicrobial secondary metabolites in the medicinal science and biotechnology industry. Different Streptomyces strains were isolated from Helianthus annuus plants and tested for antibacterial and antifungal activities. The most promising five strains were chosen for further investigation, and growth conditions for antibiotic synthesis were optimised. The supernatants were extracted in different solvents, and the extracted products were analyzed using liquid chromatography-mass spectrometry (LC-MS) and biological testing. From one of the potent strains Streptomyces globusus sp. BR123, a compound lavendamycin was identified using these analytical techniques. In addition, this potent strain also produces a strong antifungal polyene compound with a quasimolecular ion of 2072. Streptomyces sp. BR123 was genome sequenced because of its promising antimicrobial potential in order to identify the gene cluster responsible for analyzed compound “lavendamycin”. The genome analysis yielded candidate genes responsible for the production of this potent compound. The genome sequence of 8.15 Mb of Streptomyces sp. isolate BR123 with a GC content of 72.63% and 8103 protein coding genes was attained. Many antimicrobial, antiparasitic, and anticancerous compounds were detected through multiple biosynthetic gene clusters predicted by in-Silico analysis. Though, the novelty of metabolites was determined through the insignificant resemblance with known biosynthetic gene clusters. The current study gives insight into the bioactive potential of Streptomyces sp. isolate BR123 with respect to the synthesis of bioactive secondary metabolites through genomic and spectrometric analysis. Moreover, the comparative genome study revealed the connection of isolate BR123 with other Streptomyces strains, which could expand the knowledge of this genus and the mechanism involved in the discovery of new antimicrobial metabolites.

Keywords: streptomyces, secondary metabolites, genome, biosynthetic gene clusters, high performance liquid chromatography, mass spectrometry

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Authors: Meriem Abid, Erika Oliveria-Jardim, Andres Fullana, Joaquin Silvestre-Albero

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The rapid industrial development associated with the increase of volatile organic compounds (VOCs) has seriously impacted the environment. Among VOCs, hydrogen sulfide (H₂S) is known as a highly toxic, malodorous, flammable, and corrosive gas, which is emitted from diverse chemical processes, including industrial waste-gas streams, natural gas processing, and biogas purification. The high toxicity, corrosively, and very characteristic odor threshold of H2S call for urgent development of efficient desulfurization processes from the viewpoint of environmental protection and resource regeneration. In order to reduce H₂S emissions, effective technologies for have been performed. The general method of H₂S removal included amine aqueous solution, adsorption process, biological methods, and fixed-bed solid catalytic oxidation processes. Ecologically and economically, low-temperature direct oxidation of H₂S to elemental sulfur using catalytic oxidation is the preferred approach for removing H₂S-containing gas streams. A large number of catalysts made from carbon, metal oxides, clay, and others, have been studied extensively for this application. In this sense, activated carbon (AC) is an attractive catalyst for H₂S removal because it features a high specific surface area, diverse functional groups, low cost, durability, and high efficiency. It is interesting to stand out that AC is modified using metal oxides to promote the efficiency of H₂S removal and to enhance the catalytic performance. Based on these premises, the main goal of the present study is the evaluation of the H₂S adsorption performance in carbon-encapsulated iron nanoparticles obtained from an olive mill, thermally treated at 600, 800 and 1000 ºC temperatures under anaerobic conditions. These results anticipate that carbon-encapsulated iron nanoparticles exhibit a promising performance for the H₂S removal up to 360 mg/g.

Keywords: H₂S removal, catalytic oxidation, carbon encapsulated iron, olive mill wastewater

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Authors: Liang-Jen Wang, Sung-Chou Li, Yuan-Ming Yeh, Sheng-Yu Lee, Ho-Chang Kuo, Chia-Yu Yang

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Background: Dysbiosis in the gut microbial community might be involved in the pathophysiology of attention deficit/hyperactivity disorder (ADHD). The fungal component of the gut microbiome, namely the mycobiota, is a hyperdiverse group of multicellular eukaryotes that can influence host intestinal permeability. This study therefore aimed to investigate the impact of fungal mycobiome dysbiosis and intestinal permeability on ADHD. Methods: Faecal samples were collected from 35 children with ADHD and from 35 healthy controls. Total DNA was extracted from the faecal samples, and the internal transcribed spacer (ITS) regions were sequenced using high-throughput next-generation sequencing (NGS). The fungal taxonomic classification was analysed using bioinformatics tools, and the differentially expressed fungal species between the ADHD and healthy control groups were identified. An in vitro permeability assay (Caco-2 cell layer) was used to evaluate the biological effects of fungal dysbiosis on intestinal epithelial barrier function. Results: The β-diversity (the species diversity between two communities), but not α-diversity (the species diversity within a community), reflected the differences in fungal community composition between ADHD and control groups. At the phylum level, the ADHD group displayed a significantly higher abundance of Ascomycota and significantly lower abundance of Basidiomycota than the healthy control group. At the genus level, the abundance of Candida (especially Candida albicans) was significantly increased in ADHD patients compared to the healthy controls. In addition, the in vitro cell assay revealed that C. albicans secretions significantly enhanced the permeability of Caco-2 cells. Conclusions: The current study is the first to explore altered gut mycobiome dysbiosis using the NGS platform in ADHD. The findings from this study indicated that dysbiosis of the fungal mycobiome and intestinal permeability might be associated with susceptibility to ADHD.

Keywords: ADHD, fungus, gut–brain axis, biomarker, child psychiatry

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Authors: Christian Fercher, Jiaul Islam, Simon R. Corrie

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Fluorescence-based immunodiagnostics are an emerging field in biosensor development and exhibit several advantages over traditional detection methods. While various affinity biosensors have been developed to generate a fluorescence signal upon sensing varying concentrations of analytes, reagentless, reversible, and continuous monitoring of complex biological samples remains challenging. Here, we aimed to genetically engineer biosensors based on single-chain antibody fragments (scFv) that are site-specifically labeled with environmentally sensitive fluorescent unnatural amino acids (UAA). A rational design approach resulted in quantifiable analyte-dependent changes in peak fluorescence emission wavelength and enabled antigen detection in vitro. Incorporation of a polarity indicator within the topological neighborhood of the antigen-binding interface generated a titratable wavelength blueshift with nanomolar detection limits. In order to ensure continuous analyte monitoring, scFv candidates with fast binding and dissociation kinetics were selected from a genetic library employing a high-throughput phage display and affinity screening approach. Initial rankings were further refined towards rapid dissociation kinetics using bio-layer interferometry (BLI) and surface plasmon resonance (SPR). The most promising candidates were expressed, purified to homogeneity, and tested for their potential to detect biomarkers in a continuous microfluidic-based assay. Variations of dissociation kinetics within an order of magnitude were achieved without compromising the specificity of the antibody fragments. This approach is generally applicable to numerous antibody/antigen combinations and currently awaits integration in a wide range of assay platforms for one-step protein quantification.

Keywords: antibody engineering, biosensor, phage display, unnatural amino acids

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Authors: Marie Fouet, Olivier Maire, Cécile Masse, Hugues Blanchet, Salomé Coignard, Nicolas Lavesque, Guillaume Bernard

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Exacerbated by global change, extreme climatic events such as atmospheric and marine heat waves may interact with the spread of non-indigenous species and their associated impacts on marine ecosystems. Since the 1970’s, the introduction of non-indigenous species due to oyster exchanges has been numerous. Among them, the Asian date mussel Arcuatula senhousia has colonized a large number of ecosystems worldwide (e.g., California, New Zealand, Italy). In these places, A.senhousia led to important habitat modifications in the benthic compartment through physical, biological, and biogeochemical effects associated with the development of dense mussel populations. In Arcachon Bay (France), a coastal lagoon of the French Atlantic and hotspot of oyster farming, abundances of A. senhousia recently increased, following a lag time of ca. 20 years since the first record of the species in 2002. Here, we addressed the potential effects of the interaction between A. senhousia invasion and heatwave intensity on ecosystem functioning within an intertidal mudflat. More precisely, two realistic intensities (“High” and “Severe”) of combined marine and atmospheric heatwaves have been simulated in an experimental tidal mesocosm system onto which naturally varying densities of A. senhousia and associated benthic communities were exposed in sediment cores collected in situ. Following a six-day exposure, community-scale responses were assessed by measuring benthic metabolism (oxygen and nutrient fluxes) in each core. Results show that besides significantly enhanced benthic metabolism with increasing heatwave intensity, mussel density clearly mediated the magnitude of the community-scale response, thereby highlighting the importance of understanding the interactive effects of environmental stressors co-occurring with non-indigenous species and their dependencies for a better assessment of their impacts.

Keywords: arcuatula senhousia, benthic habitat, ecosystem functioning, heatwaves, metabolism

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Authors: Yunfeng Shan, Xiaoliang Wang, Youjun Zhou

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Whole-genome data from two lungfish species, along with other species, present a valuable opportunity to re-investigate the longstanding debate regarding the evolutionary relationships among tetrapods, lungfishes, and coelacanths. However, the use of bootstrap support has become outdated for large-scale phylogenomic data. Without robust phylogenetic support, the phylogenetic trees become meaningless. Therefore, it is necessary to re-evaluate the phylogenies of tetrapods, lungfishes, and coelacanths using novel measures of phylogenetic support specifically designed for phylogenomic data, as the previous phylogenies were based on 100% bootstrap support. Our findings consistently provide strong evidence favoring lungfish as the closest living relative of tetrapods. This conclusion is based on high internode certainty, relative gene support, and high gene concordance factor. The evidence stems from five previous datasets derived from lungfish transcriptomes. These results yield fresh insights into the three hypotheses regarding the phylogenies of tetrapods, lungfishes, and coelacanths. Importantly, these hypotheses are not mere conjectures but are substantiated by a significant number of genes. Analyzing real biological data further demonstrates that the inclusion of additional taxa leads to more diverse tree topologies. Consequently, gene trees and species trees may not be identical even when whole-genome sequencing data is utilized. However, it is worth noting that many gene trees can accurately reflect the species tree if an appropriate number of taxa, typically ranging from six to ten, are sampled. Therefore, it is crucial to carefully select the number of taxa and an appropriate outgroup, such as slow-evolving species, while excluding fast-evolving taxa as outgroups to mitigate the adverse effects of long-branch attraction and achieve an accurate reconstruction of the species tree. This is particularly important as more whole-genome sequencing data becomes available.

Keywords: novel measures of phylogenetic support for phylogenomic data, gene concordance factor confidence, relative gene support, internode certainty, origin of tetrapods

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Authors: C. Mayuren, C. K. Paul Wang, K. Purushotham, C. Dinesh Kumar

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Diabetes Mellitus (DM) is one of the most common non-communicable diseases globally. Although significant advances have led to better understanding of the condition and the development of effective therapies and preventive strategies, the pathway to cure remains elusive and DM prevails as a serious medical challenge in the 21st century. Oxidative stress has been suggested to contribute to the progression and pathophysiological conditions of diabetes. Madecassoside (MA) a major pentacyclic triterpenoid, has been demonstrated to possess various biological activities. However, no attempt has been made to study the antioxidant activity in diabetic rats. Therefore, the present study is aimed to evaluate the antioxidant effect of MA on streptozotocin-nicotinamide induced type-2 diabetes in Sprague-Dawley rats. The study protocol was approved by the institutional ethical committee prior to the conduct of research. Adult male Sprague-Dawley rats weighing 250-300 g were used in the study. The animals were rendered diabetic with a single intraperitoneal dose of streptozotocin (65 mg/kg) and nicotinamide (110 mg/kg). The diabetic animals after a stabilisation period of 14 days received various treatments (Madecassoside 50 mg/kg; Glimepiride 2.5 mg/kg) suspended in 0.5% carboxymethyl cellulose orally, for a period of 28 days. The animals fasted overnight after the last treatment were sacrificed and the pancreas, liver and kidneys were isolated. The weighted quantity of the samples of various treatments were homogenised in ice-cold condition and were subjected to lipid peroxidation, catalase and superoxide dismutase assay. The data’s obtained were subjected to statistical analysis. Diabetic rats showed significant increase in lipid peroxidation and decrease in enzymatic antioxidant levels. All the treated groups had significantly higher SOD, CAT and reduced LPO activity in the pancreas, liver and kidney. Results suggest madecassoside to have potential antioxidant effect against the diabetic model. However further investigations are necessary to study the mechanism at the cellular level.

Keywords: antioxidant, diabetes, madecassoside, nicotinamide, streptozotocin

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Authors: Desmond Okorie, Emmanuel Prince

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Availability of portable water has been a global challenge especially to the developing continents/nations such as Africa/Nigeria. The World Health Organization WHO has produced the guideline for drinking water quality GDWQ which aims at ensuring water safety from source to consumer. Portable water parameters test include physical (colour, odour, temperature, turbidity), chemical (PH, dissolved solids) biological (algae, plytoplankton). This paper discusses the use of wireless sensor networks to monitor water quality using efficient and effective sensors that have the ability to sense, process and transmit sensed data. The integration of wireless sensor network to a portable sensing device offers the feasibility of sensing distribution capability, on site data measurements and remote sensing abilities. The current water quality tests that are performed in government water quality institutions in Benue State Nigeria are carried out in problematic locations that require taking manual water samples to the institution laboratory for examination, to automate the entire process based on wireless sensor network, a system was designed. The system consists of sensor node containing one PH sensor, one temperature sensor, a microcontroller, a zigbee radio and a base station composed by a zigbee radio and a PC. Due to the advancement of wireless sensor network technology, unexpected contamination events in water environments can be observed continuously. local area network (LAN) wireless local area network (WLAN) and internet web-based also commonly used as a gateway unit for data communication via local base computer using standard global system for mobile communication (GSM). The improvement made on this development show a water quality monitoring system and prospect for more robust and reliable system in the future.

Keywords: local area network, Ph measurement, wireless sensor network, zigbee

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Authors: Claudiu Marcu, Cristina Paul, Adelina Andelescu, Corneliu Mircea Davidescu, Francisc Péter

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Heavy metal pollution has become a more serious environmental problem in the last several decades as a result of its toxicity and insusceptibility to the environment. Methods for removing metal ions from aqueous solution mainly consist of physical, chemical and biochemical procedures. Biosorption is defined as the removal of metal or metalloid species, compounds and particulates from solution by a biological material. Biosorption represents a very attractive method for the removal of toxic metal ions from aqueous effluents because it uses the ability of various biomass to bind the metal ions without the risk of releasing other toxic chemical compounds into the environment. The problem with using biomass or living cells as biosorbents is that their regeneration/reuse is often either impossible or very laborious. One of the most common chelating group found in biosorbents is the thiol group in cysteine. Therefore, we immobilized cysteine using covalent binding using glutaraldehyde as a linker on a synthetic sol-gel support obtained using 3-amino-propyl-trimetoxysilane and trimetoxysilane as precursors. The obtained adsorbents were used for removal of cadmium from aqueous solutions and the removal capacity was investigated in relation to the composition of the sol-gel hybrid composite, the loading of the biomolecule and the physical parameters of the biosorption process. In the same conditions, the bare sol-gel support without cysteine had no Cd removal effect, while the adsorbent with cysteine had an adsorption capacity up to 25.8 mg Cd/g adsorbent at pH 2.0 and 119 mg Cd/g adsorbent at pH 6.6, depending on cadmium concentration and adsorption conditions. We used atomic adsorption spectrometry to assess the cadmium concentration in the samples after the biosorbtion process. The parameters for the Freundlich and Langmuir adsorption isotherms where calculated from plotting the results of the adsorption experiments. The results for cysteine immobilization show a good loading capacity of the sol-gel support which indicates it could be used to immobilize metal binding proteins and by doing so boosting the heavy metal adsorption capacity of the biosorbent.

Keywords: biosorbtion, cadmium, cysteine covalent binding, sol-gel

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Authors: Almas Hamid, Ghazala Yaqub, Aqsa Riaz

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Qualitative validation was performed to detect the presence of persistent organic pollutants (POPs) in various wastewater samples collected from domestic sources (Askari XI housing society, Bedian road Lahore) industrial sources (PET bottles, pharmaceutical, textile) and a municipal drain (Hudiara drain) in Lahore. In addition wastewater analysis of the selected parameter was carried out. pH for wastewater samples from Askari XI, PET bottles, pharmaceutical, textile and Hudiara drain were 6.9, 6.7, 6.27, 7.18 and 7.9 respectively, within the NEQS Pakistan range that is 6-9. TSS for the respective samples was 194, 241, 254, 140 and 251 mg/L, in effluent for pet bottle industry, pharmaceutical and Hudiara drain and exceeded the NEQS Pakistan. Chemical oxygen demand (COD) for the wastewater samples was 896 mg/L, 166 mg/L, 419 mg/L, 812 mg/L and 610 mg/L respectively, all in excess of NEQS (150 mg/L). Similarly the biological oxygen demand (BOD) values (110.8, 170, 423, 355 and 560 mg/L respectively) were also above NEQS limits (80 mg/L). Chloride (Cl-) content, total dissolved solids (TDS) and temperature were found out to be within the prescribed standard limits. The POPs selected for analysis included five pesticides/insecticides (D. D, Karate, Commando, Finis insect killer, Bifenthrin) and three polycyclic aromatic hydrocarbons (PAHs) (naphthalene, anthracene, phenanthrene). Peak values of standards were compared with that of wastewater samples. The results showed the presence of D.D in all wastewater samples, pesticide Karate was identified in Askari XI and textile industry sample. Pesticide Commando, Finis (insect killer) and Bifenthrin were detected in Askari XI and Hudiara drain wastewater samples. In case of PAHs; naphthalene was identified in all the five wastewater samples whereas anthracene and phenanthrene were detected in samples of Askari XI housing society, PET bottles industry, pharmaceutical industry and textile industry but totally absent in Hudiara drain wastewater. Practical recommendations have been put forth to avoid hazardous impacts of incurred samples.

Keywords: HPLC studies, lahore, physicochemical analysis, wastewater

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Authors: Cristian Soitu, Alexander Feuerborn, Cyril Deroy, Alfonso Castrejon-Pita, Peter R. Cook, Edmond J. Walsh

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Microfluidics now stands as an academically mature technology after a quarter of a century research activities have delivered a vast array of proof of concepts for many biological workflows. However, translation to industry remains poor, with only a handful of notable exceptions – e.g. digital PCR, DNA sequencing – mainly because of biocompatibility issues, limited range of readouts supported or complex operation required. This technology exploits the domination of interfacial forces over gravitational ones at the microscale, replacing solid walls with fluid ones as building blocks for cell micro-environments. By employing only materials used by biologists for decades, the system is shown to be biocompatible, and easy to manufacture and operate. The method consists in displacing a continuous fluid layer into a pattern of isolated chambers overlaid with an immiscible liquid to prevent evaporation. The resulting fluid arrangements can be arrays of micro-chambers with rectangular footprint, which use the maximum surface area available, or structures with irregular patterns. Pliant, self-healing fluid walls confine volumes as small as 1 nl. Such fluidic structures can be reconfigured during the assays, giving the platform an unprecedented level of flexibility. Common workflows in cell biology are demonstrated – e.g. cell growth and retrieval, cloning, cryopreservation, fixation and immunolabeling, CRISPR-Cas9 gene editing, and proof-of-concept drug tests. This fluid-shaping technology is shown to have potential for high-throughput cell- and organism-based assays. The ability to make and reconfigure on-demand microfluidic circuits on standard Petri dishes should find many applications in biology, and yield more relevant phenotypic and genotypic responses when compared to standard microfluidic assays.

Keywords: fluid walls, micro-chambers, reconfigurable, freestyle

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Authors: Ketaki D. Belsare, Nicholas F. Polizzi, Lior Shtayer, William F. DeGrado

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Nature utilizes metalloproteins to perform chemical transformations with activities and selectivities that have long been the inspiration for design principles in synthetic and biological systems. The chemical reactivities of metalloproteins are directly linked to local environment effects produced by the protein matrix around the metal cofactor. A complete understanding of how the protein matrix provides these interactions would allow for the design of functional metalloproteins. The de novo computational design of proteins have been successfully used in design of active sites that bind metals like di-iron, zinc, copper containing cofactors; however, precisely designing active sites that can bind small molecule ligands (e.g., substrates) along with metal cofactors is still a challenge in the field. The de novo computational design of a functional metalloprotein that contains a purposefully designed substrate binding site would allow for precise control of chemical function and reactivity. Our research strategy seeks to elucidate the design features necessary to bind the cofactor protoporphyrin IX (hemin) in close proximity to a substrate binding pocket in a four helix bundle. First- and second-shell interactions are computationally designed to control orientation, electronic structure, and reaction pathway of the cofactor and substrate. The design began with a parameterized helical backbone that positioned a single histidine residue (as an axial ligand) to receive a second-shell H-bond from a Threonine on the neighboring helix. The metallo-cofactor, hemin was then manually placed in the binding site. A structural feature, pi-bulge was introduced to give substrate access to the protoporphyrin IX. These de novo metalloproteins are currently being tested for their activity towards hydroxylation and epoxidation. The de novo designed protein shows hydroxylation of aniline to 4-aminophenol. This study will help provide structural information of utmost importance in understanding de novo computational design variables impacting the functional activities of a protein.

Keywords: metalloproteins, protein design, de novo protein, biocatalysis

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