Search results for: antimicrobial resistance genes

Authors: Susana P. Dias, Andrea Gomes, Fernanda M. Ferreira, Marta F. Henriques

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Thymus mastichina L. is an endogenous medicinal and aromatic plant of the Mediterranean flora. It has been used empirically over the years as a natural preservative in food. Nowadays, the antimicrobial activity of its bioactive compounds, such as essential oils and extracts, has been well recognized. The main purpose of this study was to evaluate the antimicrobial effect of Thymus mastichina ethanolic and aqueous extracts on pathogens and spoilage microorganisms present in cheese during ripening. The effect that the extract type and its concentration has on the development of Staphylococcus aureus, Escherichia coli, and Yarrowia lipolytica populations during 24 hours, was studied 'in vitro' using appropriate culture media. The results achieved evidenced the antimicrobial activity of T. mastichina extracts against the studied strains, and the concentration of 2 mg/mL (w/v) was selected and used directly on the cheese surface during ripening. In addition to the microbiological evaluation in terms of total aerobic bacteria, Enterobacteriaceae, yeasts (particularly Y. lipolytica) and molds, the treated cheeses physicochemical evaluation (humidity, aw, pH, colour, and texture) was also performed. The results were compared with cheeses with natamicyn (positive control) and without any treatment (negative control). The physicochemical evaluation showed that the cheeses treated with ethanolic extract of Thymus mastichina, except the fact that they lead to a faster water loss during ripening, did not present considerable differences when compared to controls. The study revealed an evident antimicrobial power of the extracts, although less effective than the one shown by the use of natamycin. For this reason, the improvement of the extraction methods and the adjustment of the extract concentrations will contribute to the use of T. mastichina as a healthier and eco-friendly alternative to natamycin, that is also more attractive from an economic point of view.

Keywords: antimicrobial activity, cheese, ethanolic extract, Thymus mastichina

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Authors: M. Moaeen-ud-Din, G. Bilal, M. Yaqoob

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Identification of genes of importance regarding production traits in buffalo is impaired by a paucity of genomic resources. Choice to fill this gap is to exploit data available for cow. The cross-species application of comparative genomics tools is potential gear to investigate the buffalo genome. However, this is dependent on nucleotide sequences similarity. In this study gene diversity between buffalo and cattle was determined by using 86 gene orthologues. There was about 3% difference in all genes in term of nucleotide diversity; and 0.267±0.134 in amino acids indicating the possibility for successfully using cross-species strategies for genomic studies. There were significantly higher non synonymous substitutions both in cattle and buffalo however, there was similar difference in term of dN – dS (4.414 vs 4.745) in buffalo and cattle respectively. Higher rate of non-synonymous substitutions at similar level in buffalo and cattle indicated a similar positive selection pressure. Results for relative rate test were assessed with the chi-squared test. There was no significance difference on unique mutations between cattle and buffalo lineages at synonymous sites. However, there was a significance difference on unique mutations for non synonymous sites indicating ongoing mutagenic process that generates substitutional mutation at approximately the same rate at silent sites. Moreover, despite of common ancestry, our results indicate a different divergent time among genes of cattle and buffalo. This is the first demonstration that variable rates of molecular evolution may be present within the family Bovidae.

Keywords: buffalo, cattle, gene diversity, molecular evolution

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Authors: Masoud Soltanialvar, Ali Bagherpour

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Documented cases of human infection with H9N2 avian influenza viruses, first detected in 1999 in Hong Kong and China, indicate that these viruses can be directly transmitted from birds to humans. In this study, we characterized the mutation in the Hemagglutinin (HA) genes and proteins that correlates with a shift in affinity of the Hemagglutinin (HA) protein from the “avian” type sialic receptors to the “human” type in 10 Iranian isolates. We delineated the genomes and receptor binding profile of HA gene of some field isolates and established their phylogenetic relationship to the other Asian H9N2 sub lineages. A total of 1200 tissue samples collected from 40 farms located in various states of Iran during 2008 – 2010 as part of a program to monitor Avian Influenza Viruses (AIV) infection. To determine the genetic relationship of Iranian viruses, the Hemagglutinin (HA) genes from ten isolates were amplified and sequenced (by RT-PCR method). Nucleotide sequences (orf) of the (HA) genes were used for phylogenetic tree construction. Deduced amino acid sequences showed the presence of L226 (234 in H9 numbering) in all ten Iranian isolates which indicates a preference to binding of α (2–6) sialic acid receptors, so these Iranian H9N2 viruses have the potential to infect human beings. These isolates showed high degree of homology with 2 human H9N2 isolates A/HK/1073/99, A/HK/1074/99. Phylogenetic analysis of showed that all the HA genes of the Iranian H9N2 viruses fall into a single group within a G1-like sublineage which had contributed as donor of six internal genes to H5N1 highly pathogenic avian influenza. The results of this study indicated that all Iranian viruses have the potential to emerge as highly pathogenic influenza virus, and considering the homology of these isolates with human H9N2 strains, it seems that the potential of these avian influenza isolates to infect human should not be overlooked.

Keywords: influenza virus, hemagglutinin, neuraminidase, Iran

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Authors: Andrey V. Khromov, Antonida V. Makhotenko, Ekaterina A. Snigir, Svetlana S. Makarova, Natalia O. Kalinina, Valentin V. Makarov, Mikhail E. Taliansky

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Due to its simplicity, CRISPR/Cas9 has become widely used and capable of inducing mutations in the genes of organisms of various kingdoms. The aim of this work was to develop applications for the efficient modification of DNA coding sequences of phytoene desaturase (PDS), coilin and vacuolar invertase (Solanum tuberosum) genes, and to develop a new nanoparticles carrier efficient technology to deliver the CRISPR/Cas9 system for editing the plant genome. For each of the genes - coilin, PDS and vacuolar invertase, five single RNA guide (sgRNAs) were synthesized. To determine the most suitable nanoplatform, two types of NP platforms were used: magnetic NPs (MNPS) and gold NPs (AuNPs). To test the penetration efficiency, they were functionalized with fluorescent agents - BSA * FITS and GFP, as well as labeled Cy3 small-sized RNA. To measure the efficiency, a fluorescence and confocal microscopy were used. It was shown that the best of these options were AuNP - both in the case of proteins and in the case of RNA. The next step was to check the possibility of delivering components of the CRISPR/Cas9 system to plant cells for editing target genes. AuNPs were functionalized with a ribonucleoprotein complex consisting of Cas9 and corresponding to target genes sgRNAs, and they were biolistically bombarded to axillary buds and apical meristems of potato plants. After the treatment by the best NP carrier, potato meristems were grown to adult plants. DNA isolated from this plants was sent to a preliminary fragment of the analysis to screen out the non-transformed samples, and then to the NGS. The present work was carried out with the financial support from the Russian Science Foundation (grant No. 16-16-04019).

Keywords: biobombardment, coilin, CRISPR/Cas9, nanoparticles, NPs, PDS, sgRNA, vacuolar invertase

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Authors: Mahmoud H. Abu Elella, Riham R. Mohamed, Magdy W. Sabaa

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Introduction: Microbial pollution is global problem threatening the human health. It is resulted by pathogenic microorganisms such as Escherichia coli (E. coli), Staphylococcus aureus (S. aureus) and other pathogenic strains. They cause a dangerous effect on human health, so great efforts have been exerted to produce new and effective antimicrobial agents. Nowadays, natural polysaccharides, such as chitosan and its derivatives are used as antimicrobial agents. The aim of our work is to synthesize of a biodegradable polymer such as N-quaternized chitosan (NQC) then Characterization of NQC by using different analysis techniques such as Fourier transform infrared (FTIR) and Scanning electron microscopy (SEM) and using it as an antibacterial agent against different pathogenic bacteria. Methods: Synthesis of NQC using dimethylsulphate. Results: FTIR technique exhibited absorption peaks of NQC, SEM images illustrated that surface of NQC was smooth and antibacterial results showed that NQC had a high antibacterial effect. Discussion: NQC was prepared and it was proved by FTIR technique and SEM images antibacterial results exhibited that NQC was an antibacterial agent.

Keywords: antimicrobial agent, N-quaternized chitosan chloride, silver nanocomposites, sodium polyacrylate

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Authors: Carolina Gutierrez-Cortes, Hector Suarez, Gustavo Buitrago

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Bacteriocins are antimicrobial peptides produced by bacteria as a competitive strategy for substrate and habitat. Those peptides have a potential use as food biopreservatives due to their antimicrobial activity against foodborne pathogens, avoiding the use of additives that can be harmful to consumers. The industrial production of bacteriocins is currently expensive; one of the options to be competitive is the development of economic culture media, for example, with the use of agro-industrial wastes such as whey. This study evaluated the growth and production of bacteriocins from four strains: Pediococcus pentosaceus 63, Pediococcus pentosaceus 145, Pediococcus pentosaceus 146 and Pediococcus pentosaceus 147 isolated from ‘minas cheese’ (artisanal cheese made from raw milk in the state of Minas Gerais, Brazil) in order to select a strain with growth at high rates and higher antimicrobial activity against Listeria monocytogenes 104 after incubation on the culture medium designed with whey and other components. The media used were: MRS broth, modified MRS broth (using different sources of carbon and nitrogen and different amounts of micronutrients) and a culture medium designed by a factorial design using whey and other components. The final biomass concentrations of the four strains in MRS broth after 24 hours of incubation were very similar 9.25, 9.33, 9.25 and 9.22 (log CFU/mL) for P. pentosaceus 63, P. pentosaceus 145, P. pentosaceus 146 and P. pentosaceus 147 respectively. In the same assays, antimicrobial activity of 3200 AU/mL for the first three and of 12800 AU/mL for P. pentosaceus 147 were obtained. Culture of P. pentosaceus 63 on modified MRS broth, showed the effect of some sources of carbon on the activity of bacteriocin, obtaining 12800 AU/mL with dextrose and 25600 AU/mL with maltose. Cultures of P. pentosaceus 145, 146 and 147 with these same sugars presented activity of 12800 AU/mL. It was observed that the modified MRS medium using whey increased the antimicrobial activity of the strains at 16000, 6400, 16000 and 19200 AU/mL for each strain respectively, keeping the biomass at values close to 9 log units. About nitrogen sources, it was observed that the combination of peptone (10 g /L), meat extract (10 g/L) and yeast extract (5 g/L) promoted the highest activity (12800 AU/mL), and in all cases MgSO4, MnSO4, K2HPO4 and ammonium citrate at low concentrations adversely affected bacteriocin production. Because P. pentosaceus 147 showed the highest antimicrobial activity in the presence of whey, it was used to evaluate the culture medium (peptone (10 g/L), meat extract (8 g/L), yeast extract (2 g/L), Tween® 80 (1 g/L), ammonium citrate (2 g/L), sodium acetate (5 g/L), MgSO4 (0.2 g/L), MnSO4 (0.04 g/L)). With the designed medium added with whey, 9.34 log units of biomass concentration and 19200 AU/mL were achieved for P. pentosaceus 147. The above suggest that the new medium promotes the antimicrobial activity of P. pentosaceus 147 allowing the use of an economic medium using whey.

Keywords: antimicrobial activity, bacteriocins, pediococcus, whey

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Authors: Dawid Sołoducha, Tomasz Borowski, Agata Markowska-Szczupak, Aneta Wesołowska, Marian Kordas, Rafał Rakoczy

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Many studies report varied effects of the magnetic field in medicine, but applications are still missing. Also, essential oils (EOs) were historically used in healing therapies, food preservation and the cosmetic industry due to their wound healing and antioxidant properties and antimicrobial activity. Unfortunately, the chemical characterization of EOs activates its antibacterial action only at a fairly high concentration. They can cause skin reactions, e.g., irritation (irritant contact dermatitis) or allergic contact dermatitis; therefore, they should always be used with caution. However, the administration of EOs to achieve the desired antimicrobial activity and stability with long-term medical usage in low concentration is challenging. The aim of this work was to investigate the antimicrobial activity of commercial Pinus sylvestris L. essential oil from Polish company Avicenna-Oil® under Rotating Magnetic Field (RMF) at f = 1 – 50 Hz. The novel construction of the magnetically assisted self-constructed reactor (MAP) was applied for this study. The chemical composition of essential pine oil was determined by gas chromatography coupled with mass spectrometry (GC-MS). Model bacteria Escherichia coli K12 (ATCC 25922) was used. Different concentrations of pine oil was prepared: 100% 50%, 25%, 12.5% and 6.25%. The disc diffusion and MIC test were done. To examine the effect of essential pine oil and rotating magnetic field RMF on antibacterial performance agar plate method was used. Pine oil consist of α-pinene (28.58%), β-pinene (17.79%), δ-3-carene (14.17%) and limonene (11.58%). The present study indicates the exposition to the RMF, as compared to the unexposed controls causing an increase in the efficacy of antibacterial properties of pine oil. We have shown that the rotating magnetic fields (RMF) at a frequency, f, between 25 Hz to 50 Hz, increase the antimicrobial efficiency of oil at lower than 50% concentration. The new method can be applied in many fields e.g. aromatherapy, medicine as a component of dressing, or as food preservatives.

Keywords: rotating magnetic field, pine oil, antimicrobial activity, Escherichia coli

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Authors: R. Frikha, H. Kamoun

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Background:Chronicmyeloidleukemia (CML), a hematologicaldisease, ischaracterized by t (9; 22) and relatedoncogene BCR-ABL formation. Although Tyrosine kinase inhibitors (TKIs) have revolutionized the treatment of CML, resistanceoccurs and possibly médiates by mutation in severalgenesindependently of the bcr-abl1 kinase mechanism. it has been reportedthat JAK2V617F/BCR-ABL double positivitymaybe a potential marker of resistance in CML. Aims: This studywasinvestigated the JAK2V617F mutation in TKI-resistant CML patients. Methods: A retrospectivestudywasconducted in the Hospital University of Sfax, south of Tunisia, including all CML TKI-resistant patients. A Sanger sequencingwasperformedusing a high-fidelity DNA polymerase. Results:Nineresistant CP-CML patients wereenrolled in thisstudy. The JAK2V617F mutation wasdetectedin 3 patients with TKI resistance. Conclusion:Despite the limit of ourstudy, ourfinding highlights the high frequency of JAK2V617F/BCR-ABL double positivity as an important marker of resistance. So; the combination of JAK and TKI inhibitorsmightbe effective and potentiallybeguided by molecular monitoring of minimal residual disease1.

Keywords: chronic myeloid leukemia, tyrosine kinase inhibitors, resistance, JAK2V617F, BCR-ABL

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Authors: D. J. Kalita

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Cancer is one of the prime causes of early death worldwide. Mutation of the gene involve in DNA repair and damage, like BRCA2 (Breast cancer gene two) genes, can be detected efficiently by PCR-RFLP to early breast cancer diagnosis and adopt the suitable method of treatment. Host Defense Peptide can be used as blueprint for the design and synthesis of novel anticancer drugs to avoid the side effect of conventional chemotherapy and chemo resistance. The change at nucleotide position 392 of a -› c in the cancer sample of dog mammary tumour at BRCA2 (exon 7) gene lead the creation of a new restriction site for SsiI restriction enzyme. This SNP may be a marker for detection of canine mammary tumour. Support vector machine (SVM) algorithm was used to design and predict the anticancer peptide from the mature functional peptide. MTT assay of MCF-7 cell line after 48 hours of post treatment showed an increase in the number of rounded cells when compared with untreated control cells. The ability of the synthesized peptide to induce apoptosis in MCF-7 cells was further investigated by staining the cells with the fluorescent dye Hoechst stain solution, which allows the evaluation of the nuclear morphology. Numerous cells with dense, pyknotic nuclei (the brighter fluorescence) were observed in treated but not in control MCF-7 cells when viewed using an inverted phase-contrast microscope. Thus, PCR-RFLP is one of the attractive approach for early diagnosis, and synthetic cationic peptide can be used for the treatment of canine mammary tumour.

Keywords: cancer, cationic peptide, host defense peptides, Breast cancer genes

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Authors: Rudzani Manafe, Ezekiel Green

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Brucella, has many different virulence factors that act as a causative agent of brucellosis, depending on the environment and other factors, some factors may play a role more than others during infection and as a result, play a role in becoming a causative agent for pathogenesis. Brucella melitensis and Brucella abortus are considered to be pathogenic to humans. The genetic regularity of nine potential causes of virulence of two Brucella species in Eastern Cape livestock have been examined. A hundred and twenty isolates obtained from Molecular Pathogenesis and Molecular Epidemiology Research Group (MPMERG) were used for this study. All isolates were grown on Brucella agar medium. Nine primer pairs were used for the detection of virB2, virB5, vceC, btpA, btpB, prpA, betB, bpe275, and bspB virulence factors using Polymerase chain reaction (PCR). Approximately 100% was observed for genes BecC and BetB from B. arbotus. While the lowest gene observed was PrpA at 4.6% from B. arbotus. BetB was detected in 34.7%, while virB2 and prpA (0%) were not detected in B. melitensis. The results from this research suggest that most isolates of Brucella have virulence-related genes associated with disease pathogenesis. Finally, our findings showed that Brucella strains in the Eastern Cape Province are extremely virulent as virulence characteristics exist in most strains investigated.

Keywords: putative virulence genes, brucella, polymerase chain reaction, milk

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Authors: Sheila M. Wicks, Gail Mahady, Udesh Patel, Joanna Michel, Armando Caceres

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Introduction: The genus piperaceae contains approximately 1000 species of herbs scrubs small trees and hanging vines distributed in both hemispheres. During our ethno medical work in Guatemala of the 27 plant families documented for us e by the Qeqchi Maya for reproductive disorders the most prominent were the Piperaceae (15%) and Menispermiaceae. Our Previous work showed that extracts from form Piper and Cissampelos species bound to both and progesterone and the estrogen receptors. In this work active extracts from Piper aeruginosibaccum Trelease, P auritum, P tuerckheimii and Cissampels tropaeolifolia were tested in functionalized cell based assays including a SEAP reporter gene and by qPCR of ER-responsive gene expression in MCF-7cells. In the reporter gene assay P aeruginosibaccum was estrogenic and enhanced E2 EFFECTS IN MCF-7 CELLS. P. tuerckheimi was not estrogenic alone but significantly enhanced the effects of E2 on SEAP reporter gene expression. Both altered mRNA expression of E2 responsive genes in MCF-7. Methods: this is collaborative project between University of Illinois at Chicago and University of San Carlos Guatemala City. 144 spices of plants were collected in Guatemala of which 57 used to treat a variety of women's reproductive health. The Genus Piperaraceae contains approximately 1000 species of herbs scrubs and small trees. Active extracts of the plants were tested in functionalized in cell-based bioassays including SEAP reporter genes. Results demonstrated altered mRNA expression of E2 responsive genes in MC-7 cells plants were collected in Guatemala of which 57 used. Conclusion of the 5 plants tested all were shown to contain components of binding to estrogenic receptor to a greater or lesser degree. These effects support the use of QEqchi Maya women in Guatemala for reproductive.

Keywords: reporter genes, MC7, guatemala piperaceae, reproductive health

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Authors: Farideh Halakou, Emel Sen, Attila Gursoy, Ozlem Keskin

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Protein–Protein Interactions (PPIs) mediate major biological processes in living cells. The study of PPIs as networks and analyze the network properties contribute to the identification of genes and proteins associated with diseases. In this study, we have created the sub-networks of brain and lung metastasis from primary tumor in breast cancer. To do so, we used seed genes known to cause metastasis, and produced their interactions through a network-topology based prioritization method named GUILDify. In order to have the experimental support for the sub-networks, we further curated them using STRING database. We proceeded by modeling structures for the interactions lacking complex forms in Protein Data Bank (PDB). The functional enrichment analysis shows that KEGG pathways associated with the immune system and infectious diseases, particularly the chemokine signaling pathway, are important for lung metastasis. On the other hand, pathways related to genetic information processing are more involved in brain metastasis. The structural analyses of the sub-networks vividly demonstrated their difference in terms of using specific interfaces in lung and brain metastasis. Furthermore, the topological analysis identified genes such as RPL5, MMP2, CCR5 and DPP4, which are already known to be associated with lung or brain metastasis. Additionally, we found 6 and 9 putative genes that are specific for lung and brain metastasis, respectively. Our analysis suggests that variations in genes and pathways contributing to these different breast metastasis types may arise due to change in tissue microenvironment. To show the benefits of using structural PPI networks instead of traditional node and edge presentation, we inspect two case studies showing the mutual exclusiveness of interactions and effects of mutations on protein conformation which lead to different signaling.

Keywords: breast cancer, metastasis, PPI networks, protein conformational changes

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Authors: Dong-Hyun Kim, Wan-Ho Chung, Hak-Sung Kim

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In this work, non-destructive evaluation was conducted to measure the sheet resistance of silver nanowire coated film and find a damage of that film using terahertz (THz) wave. Pulse type THz instrument was used, and the measurement was performed under transmission and pitch-catch reflection modes with 30 degree of incidence angle. In the transmission mode, the intensity of the THz wave was gradually increased as the conductivity decreased. Meanwhile, the intensity of THz wave was decreased as the conductivity decreased in the pitch-catch reflection mode. To confirm the conductivity of the film, sheet resistance was measured by 4-point probe station. Interaction formula was drawn from a relation between the intensity and the sheet resistance. Through substituting sheet resistance to the formula and comparing the resultant value with measured maximum THz wave intensity, measurement of sheet resistance using THz wave was more suitable than that using 4-point probe station. In addition, the damage on the silver nanowire coated film was detected by applying the THz image system. Therefore, the reliability of the entire film can be also be ensured. In conclusion, real-time monitoring using the THz wave can be applied in the transparent electrodes with detecting the damaged area as well as measuring the sheet resistance.

Keywords: terahertz wave, sheet resistance, non-destructive evaluation, silver nanowire

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Authors: Haroon Jamshaid Qazi, Namrah Wahid, Sanaullah Iqbal, Raheel Suleman

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The aim of the current study was to evaluate the antioxidant and antimicrobial potential of Rutin in cheddar cheese. The study was conducted by adding the Rutin in the cheddar cheese in different concentrations according to experimental design, i.e., T1 (20 ppm Rutin), T2 (40 ppm Rutin), T3 (60 ppm Rutin), T4 (80 ppm Rutin). BHT was taken as a positive control at a concentration of 200 ppm, and negative control had neither Rutin nor BHT. The ripening time for cheeses was 90 days at a temperature of 8°C. The results of the various antioxidants assays (Total phenolic contents (TPC) and Antioxidant activity (AA), with storage stability tests (Anisidine value (AV) and Thiobarbituric acid value (TBARS)) performed during different storage intervals 0, 30, 60 and 90 days exhibited that AA in linoleic acid and TPC were significantly (p < 0.05) increased by the addition of rutin to cheese at all concentrations. Moreover, significant reduction in the TBARS values was also observed during the storage period. Rutin also showed a good potential to inhibit the microbial proliferation in the treated samples of cheese. There was a significant decreasing trend seen in total plate count and yeasts and molds count. The sensorial attributes i.e., color, flavor, odor and overall acceptability were increased after adding Rutin to cheddar cheese.

Keywords: cheddar cheese, Rutin, antioxidant, antimicrobial

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Authors: M. Marjaneh, M. Y. Narazah, H. Shahrul

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Array-based gene expression analysis is a powerful tool to profile expression of genes and to generate information on therapeutic effects of new anti-cancer compounds. Anti-apoptotic effect of thymoquinone was studied in MCF7 breast cancer cell line using gene expression profiling with cDNA micro array. The purity and yield of RNA samples were determined using RNeasyPlus Mini kit. The Agilent RNA 6000 Nano LabChip kit evaluated the quantity of the RNA samples. AffinityScript RT oligo-dT promoter primer was used to generate cDNA strands. T7 RNA polymerase was used to convert cDNA to cRNA. The cRNA samples and human universal reference RNA were labelled with Cy-3-CTP and Cy-5-CTP, respectively. Feature Extraction and GeneSpring software analysed the data. The single experiment analysis revealed involvement of 64 pathways with up-regulated genes and 78 pathways with down-regulated genes. The MAPK and p38-MAPK pathways were inhibited due to the up-regulation of PTPRR gene. The inhibition of p38-MAPK suggested up-regulation of TGF-ß pathway. Inhibition of p38 - MAPK caused up-regulation of TP53 and down-regulation of Bcl2 genes indicating involvement of intrinsic apoptotic pathway. Down-regulation of CARD16 gene as an adaptor molecule regulated CASP1 and suggested necrosis-like programmed cell death and involvement of caspase in apoptosis. Furthermore, down-regulation of GPCR, EGF-EGFR signalling pathways suggested reduction of ER. Involvement of AhR pathway which control cytochrome P450 and glucuronidation pathways showed metabolism of Thymoquinone. The findings showed differential expression of several genes in apoptosis pathways with thymoquinone treatment in estrogen receptor-positive breast cancer cells.

Keywords: cDNA microarray, thymoquinone, CARD16, PTPRR, CASP10

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Authors: Genesis Julyus T. Agcaoili, Esperanza C. Cabrera

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Thirty (30) isolates of lactic acid bacteria (LAB) from traditionally-prepared fermented products specifically fermented soy-bean paste, fermented mustard and fermented rice-fish mixture were studied for their in vitro antimicrobial and cytotoxic activities. Seventeen (17) isolates were identified as Lactobacillus plantarum, while 13 isolates were identified as Enterococcus spp using 16s rDNA sequences. Disc diffusion method was used to determine the antibacterial activity of LAB against Staphylococcus aureus (ATCC 25923) and Escherichia coli (ATCC 25922), while the modified agar overlay method was used to determine the antifungal activity of LAB isolates on the yeast Candida albicans, and the dermatophytes Microsporum gypseum, Trichophyton rubrum and Epidermophyton floccosum. The filter-sterilized LAB supernatants were evaluated for their cytotoxicity to mammalian colon cancer cell lines (HT-29 and HCT116) and normal human dermal fibrolasts (HDFn) using resazurin assay (PrestoBlueTM). Colchicine was the positive control. No antimicrobial activity was observed against the bacterial test organisms and the yeast Candida albicans. On the other hand, all of the tested LAB strains were fungicidal for all the test dermatophytes. Cytotoxicity index profiles of the supernatants of the 15 randomly picked LABs and negative control (brain heart infussion broth) suggest nontoxicity to the cells when compared to colchicine, whereas all LAB supernatants were found to be cytotoxic to HT-29 and HCT116 colon cancer cell lines. Results provide strong support for the role of the lactic acid bacteria studied in antimicrobial treatment and anticancer therapy.

Keywords: antimicrobial, fermented products, fungicidal activity, lactic acid bacteria, probiotics

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Authors: A. Nusrath Unissa, Sameer Hassan, Luke Elizabeth Hanna

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Altered drug binding may be an important factor in isoniazid (INH) resistance, rather than major changes in the enzyme’s activity as a catalase or peroxidase (KatG). The identification of structural or functional defects in the mutant KatGs responsible for INH resistance remains as an area to be explored. In this connection, the differences in the binding affinity between wild-type (WT) and mutants of KatG were investigated, through the generation of three mutants of KatG, Ser315Thr [S315T], Ser315Asn [S315N], Ser315Arg [S315R] and a WT [S315]) with the help of software-MODELLER. The mutants were docked with INH using the software-GOLD. The affinity is lower for WT than mutant, suggesting the tight binding of INH with the mutant protein compared to WT type. These models provide the in silico evidence for the binding interaction of KatG with INH and implicate the basis for rationalization of INH resistance in naturally occurring KatG mutant strains of Mycobacterium tuberculosis.

Keywords: Mycobacterium tuberculosis, KatG, INH resistance, mutants, modelling, docking

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Authors: Radhika S. Oke, Rebecca S. Thombre

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Medicinal plants and herbs have a great history of their utility as remedy for treatment of variety of ailments. Secondary metabolites present in these plants are responsible for their medicinal activity. In the present investigation, phytochemical screening of aqueous and alcoholic leaf extract of Anethum graveolens L. was performed. Total phenolic content and total antioxidant activity of the extracts was quantitatively estimated by Folin-Ciocalteau method and DPPH (1, 1-Diphenyl-2-picryl hydrazyl) method respectively. Qualitative tests suggested that Alkaloids, tannins and phenolic compounds were present in all the extracts of the plant. Aqueous extracts was found to have more phytochemicals as compared to alcoholic extracts. Extract of Anethum graveolens L. was found to contain good amount phenolics and exhibited antioxidant activity. The extracts also demonstrated potent antimicrobial activity against selected gram positive and negative bacteria. The study revealed the potential application of Anethum graveolens L. (Dill) in medicine and health.

Keywords: Anethum graveolens L., antioxidant, antimicrobial activity, medicine and health

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Authors: Al-Bogami Abdullah Saad

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We reported the environmental benign synthesis of chalcones, 2-pyrazolines and cyclohexanones under microwave irradiation. Chalcones were obtained by the condensation of each of 2-hydroxyacetophenone derivatives with α-naphthaldehyde under microwave irradiation. The condensation reactions of each of synthesized chalcones with phenyl hydrazine under microwave irradiation in the presence of dry acetic acid as a cyclizing agent gave 2-pyrazolines. Also, the new cyclohexenone derivatives, valuable intermediates to synthesize fused heterocycles, have been prepared by the cyclocondensation of each of hydroxychalcones with ethyl acetoacetate. The structures of the synthesized compounds were elucidated by Infrared (IR) spectrometry, Nuclear Magnetic Resonance (NMR), Mass Spectrometry(MS) and elmental analysis. The results indicate that unlike classical heating, microwave irradiation results in higher yields with shorter and cleaner reactions. The synthesized compounds were screened for antimicrobial activity against Staphylococcus aureus, Escherichia coli, Candida Albicans and Aspergillus niger. We clarified the effects of different substituents in the tested compounds on the obtaind antibacterial activities and antifungal activities.

Keywords: microwave irradiation, 2-Hydroxyacetophenone, α-Naphthaldehyde, pyrazoline, cyclohexenone, antimicrobial activity

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Authors: Ankan Roy, Niharika, Samir Kumar Patra

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Anomalous nexus of complex topological assemblies and spatiotemporal epigenetic choreography at chromosomal territory may forms the most sophisticated regulatory layer of gene expression in cancer. Colon cancer is one of the leading malignant neoplasms of the lower gastrointestinal tract worldwide. There is still a paucity of information about the complex molecular mechanisms of colonic cancerogenesis. Bioinformatics prediction and analysis helps to identify essential genes and significant pathways for monitoring and conquering this deadly disease. The present study investigates and explores potential hub genes as biomarkers and effective therapeutic targets for colon cancer treatment. Colon cancer patient sample containing gene expression profile datasets, such as GSE44076, GSE20916, and GSE37364 were downloaded from Gene Expression Omnibus (GEO) database and thoroughly screened using the GEO2R tool and Funrich software to find out common 2 differentially expressed genes (DEGs). Other approaches, including Gene Ontology (GO) and KEGG pathway analysis, Protein-Protein Interaction (PPI) network construction and hub gene investigation, Overall Survival (OS) analysis, gene correlation analysis, methylation pattern analysis, and hub gene-Transcription factors regulatory network construction, were performed and validated using various bioinformatics tool. Initially, we identified 166 DEGs, including 68 up-regulated and 98 down-regulated genes. Up-regulated genes are mainly associated with the Cytokine-cytokine receptor interaction, IL17 signaling pathway, ECM-receptor interaction, Focal adhesion and PI3K-Akt pathway. Downregulated genes are enriched in metabolic pathways, retinol metabolism, Steroid hormone biosynthesis, and bile secretion. From the protein-protein interaction network, thirty hub genes with high connectivity are selected using the MCODE and cytoHubba plugin. Survival analysis, expression validation, correlation analysis, and methylation pattern analysis were further verified using TCGA data. Finally, we predicted COL1A1, COL1A2, COL4A1, SPP1, SPARC, and THBS2 as potential master regulators in colonic cancerogenesis. Moreover, our experimental data highlights that disruption of lipid raft and RAS/MAPK signaling cascade affects this gene hub at mRNA level. We identified COL1A1, COL1A2, COL4A1, SPP1, SPARC, and THBS2 as determinant hub genes in colon cancer progression. They can be considered as biomarkers for diagnosis and promising therapeutic targets in colon cancer treatment. Additionally, our experimental data advertise that signaling pathway act as connecting link between membrane hub and gene hub.

Keywords: hub genes, colon cancer, DNA methylation, epigenetic engineering, bioinformatic predictions

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Authors: Asma Meliani, S. Lakehal, F. Z. Benrebiha, C. Chaouia

Abstract:

There is an increasing interest in phytochemicals as new source of natural antioxidant and antimicrobial agents. Plants essential oils have come more into the focus of phytomedicine. Many researchers have reported various biological and/or pharmacological properties of Artemisia herba alba Asso essential oil. The present study describes antimicrobial and antioxidant properties of Artemisia herba alba Asso essential oil. Artemisia herba alba Asso essential oil obtained by hydrodistillation (using Clevenger type apparatus) growing in Algeria (M’sila) was analyzed by GC-MS. The essential oil yield of the study was 0.7%. The major components were found to be camphor, chrysanthenone et 1,8-cineole. The antimicrobial activity of the essential oil was tested against four bacteria (Gram-negative and Gram-positive) and three fungi using the diffusion method and by determining the inhibition zone. The oil was found to have significant antibacterial activity. In addition, antioxidant activity was determined by 1, 1-diphenyl-1-picrylhydrazyl (DPPH) assay, ferric reducing (FRAP) assay and β-carotene bleaching test, and high activity was found for Artemisia herba-alba oil.

Keywords: Artemisia herba-alba, essential oil, antibacterial activity, antioxidant activity

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Authors: Anil Kumar, Diwakar Aggarwal, M. Sudhakara Reddy

Abstract:

The wood of Eucalyptus, Populus and bamboos are some important species used as raw material for the manufacture of pulp. However, higher levels of lignin pose a problem during Kraft pulping and yield of pulp is also lower. In order to increase the yield of pulp per unit wood and reduce the use of chemicals during kraft pulping it is important to reduce the lignin content and/or increase cellulose content in wood. Cellulose biosynthesis in wood takes place by the coordinated action of many enzymes. The two important enzymes are KORRIGAN and SUCROSE SYNTHASE. KORRIGAN (Endo-1,4--glucanase) is implicated in the process of editing growing cellulose chains and improvement of the crystallinity of produced cellulose, whereas SUCROSE SYNTHASE is involved in providing substrate (UDP-glucose) for growing cellulose chains. The present study was aimed at the cloning, characterization and overexpression of these genes in Eucalyptus and Populus. An efficient shoot organogenesis protocol from leaf explants taken from micro shoots of the species has been developed. Agrobacterium mediated genetic transformation using Agrobacterium tumefaciens strain EHA105 and LBA4404 harboring binary vector pBI121 was achieved. Both the genes were cloned from cDNA library of Populus deltoides. These were subsequently characterized using various bioinformatics tools. The cloned genes were then inserted into pBI121 under the CaMV35S promotors replacing GUS gene. The constructs were then mobilized into above strains of Agrobacterium and used for the transformation work. Subsequently, genetic transformation of these clones with target genes following already developed protocol is in progress. Four transgenic lines of Eucalyptus tereticornis overexpressing Korrigan gene under the strong constitutive promoters CaMV35S have been developed, which are being further evaluated. Work on development of more transgenic lines overexpressing these genes in Populus and Eucalyptus is also in progress. This presentation will focus on important developments in this direction.

Keywords: Eucalyptus tereticornis, genetic transformation, Kraft pulping Populus deltoides

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Authors: B. Nageshwar Rao

Abstract:

Urinary tract infection (UTI) represents the most commonly acquired bacterial infection worldwide, with substantial morbidity, mortality, and economic burden. The objective of the study is to characterize the microbial profiles of uropathogenic in the obstetric population by RTPCR. Study design: An observational cross-sectional study was performed at a single tertiary health care hospital among 50 pregnant women with UTIs, including asymptomatic and symptomatic patients attending the outpatient department and inpatient department of Obstetrics and Gynaecology.Methods: Serotyping and genes detection of various uropathogens were studied using RTPCR. Pulse filed gel electrophoresis methods were used to determine the various genetic profiles. Results: The present study shows that CsgD protein, involved in biofilm formation in Escherichia coli, VIM1, IMP1 genes for Klebsiella were identified by using the RTPCR method. Our results showed that the prevalence of VIM1 and IMP1 genes and CsgD protein in E.coli showed a significant relationship between strong biofilm formation, and this may be due to the prevalence of specific genes. Finally, the genetic identification of RTPCR results for both bacteria was correlated with each other and concluded that the above uropathogens were common isolates in producing Biofilm in the pregnant woman suffering from urinary tract infection in our hospital observational study.

Keywords: biofilms, Klebsiella, E.coli, urinary tract infection

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Authors: Chenyu Huang, Minrong Cui, Hua Fang, Luqing Zhang, Yunlong Yu

Abstract:

The emergence and spread of antibiotic resistance genes (ARGs) have become a pressing issue in global agricultural production. However, understanding how these ARGs spread across different spatial scales, especially when exposed to both pesticides and antibiotics, has remained a challenge. Here, metagenomic assembly and binning methodologies were used to determine the mechanism of ARG propagation within soil-lettuce systems exposed to both fungicides and antibiotics. The results of our study showed that the presence of fungicide and antibiotic stresses had a significant impact on certain bacterial communities. Notably, we observed that ARGs were primarily transferred from the soil to the plant through plasmids. The selective pressure exerted by fungicides and antibiotics contributed to an increase in unique ARGs present on lettuce leaves. Moreover, ARGs located on chromosomes and plasmids followed different transmission patterns. The presence of diverse selective pressures, a result of compound treatments involving antibiotics and fungicides, amplifies this phenomenon. Consequently, there is a higher probability of bacteria developing multi-antibiotic resistance under the combined pressure of fungicides and antibiotics. In summary, our findings highlight that combined fungicide and antibiotic treatments are more likely to drive the acquisition of ARGs within the soil-plant system and may increase the risk of human ingestion.

Keywords: soil-lettuce system, fungicide, antibiotic, ARG, transmission

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Authors: Ghibeche Abderrahmane

Abstract:

To examine whether genes associated with cellular defense against oxidative stress are associated with insulin sensitivity, patients with type 2 diabetes (n=7) and age-matched (n=5) and young (n=9) control subjects underwent a euglycemic-hyperinsulinemic clamp for 120 min. Muscle samples were obtained before and after the clamp and analyzed for heat shock protein (HSP)72 and heme oxygenase (HO)-1 mRNA, intramuscular triglyceride content, and the maximal activities of β-hyroxyacyl-CoA dehydrogenase (β-HAD) and citrate synthase (CS). Basal expression of both HSP72 and HO-1 mRNA were lower (P < 0.05) by 33 and 55%, respectively, when comparing diabetic patients with age-matched and young control subjects, with no differences between the latter groups. Both basal HSP72 (r = 0.75, P < 0.001) and HO-1 (r = 0.50,P < 0.05) mRNA expression correlated with the glucose infusion rate during the clamp. Significant correlations were also observed between HSP72 mRNA and both β-HAD (r = 0.61, P < 0.01) and CS (r = 0.65, P < 0.01). HSP72 mRNA was induced (P < 0.05) by the clamp in all groups. Although HO-1 mRNA was unaffected by the clamp in both the young and age-matched control subjects, it was increased (P < 0.05) ∼70-fold in the diabetic patients after the clamp. These data demonstrate that genes involved in providing cellular protection against oxidative stress are defective in patients with type 2 diabetes and correlate with insulin-stimulated glucose disposal and markers of muscle oxidative capacity. The data provide new evidence that the pathogenesis of type 2 diabetes involves perturbations to the antioxidant defense mechanism within skeletal muscle.

Keywords: euglycemic-hyperinsulinemic, HSP72, mRNA, diabete

Procedia PDF Downloads 415

Authors: Asma Meliani, S. Lakehal, F. Z. Benrebiha, C. Chaouia

Abstract:

There is an increasing interest in phytochemicals as new source of natural antioxidant and antimicrobial agents. Plants essential oils have come more into the focus of phytomedicine. Many researchers have reported various biological and/or pharmacological properties of Artemisia herba alba Asso essential oil. The present study describes antimicrobial and antioxidant properties of Artemisia herba alba Asso essential oil. Artemisia herba alba Asso essential oil obtained by hydrodistillation (using Clevenger type apparatus) growing in Algeria (M’sila) was analyzed by GC-MS. The essential oil yield of the study was 0.7 %. The major components were found to be camphor, chrysanthenone et 1,8-cineole. The antimicrobial activity of the essential oil was tested against four bacteria (Gram-negative and Gram-positive) and one fungi using the diffusion method and by determining the inhibition zone. The oil was found to have significant antibacterial activity. In addition, antioxidant activity was determined by 1,1-diphenyl-1-picrylhydrazyl (DPPH) assay, ferric reducing (FRAP) assay and β-carotene bleaching test, and high activity was found for Artemisia herba-alba oil.

Keywords: Artemisia herba-alba, essential oil, antibacterial activity, antioxidant activity

Procedia PDF Downloads 445

Authors: Hala Alshamlan, Ghada Badr, Yousef Alohali

Abstract:

Cancer is one of the dreadful diseases, which causes considerable death rate in humans. DNA microarray-based gene expression profiling has been emerged as an efficient technique for cancer classification, as well as for diagnosis, prognosis, and treatment purposes. In recent years, a DNA microarray technique has gained more attraction in both scientific and in industrial fields. It is important to determine the informative genes that cause cancer to improve early cancer diagnosis and to give effective chemotherapy treatment. In order to gain deep insight into the cancer classification problem, it is necessary to take a closer look at the proposed gene selection methods. We believe that they should be an integral preprocessing step for cancer classification. Furthermore, finding an accurate gene selection method is a very significant issue in a cancer classification area because it reduces the dimensionality of microarray dataset and selects informative genes. In this paper, we classify and review the state-of-art gene selection methods. We proceed by evaluating the performance of each gene selection approach based on their classification accuracy and number of informative genes. In our evaluation, we will use four benchmark microarray datasets for the cancer diagnosis (leukemia, colon, lung, and prostate). In addition, we compare the performance of gene selection method to investigate the effective gene selection method that has the ability to identify a small set of marker genes, and ensure high cancer classification accuracy. To the best of our knowledge, this is the first attempt to compare gene selection approaches for cancer classification using microarray gene expression profile.

Keywords: gene selection, feature selection, cancer classification, microarray, gene expression profile

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Authors: Nassima Behidj-Benyounes, Thoraya Dahmene, Khaled Benyounes Nadjiba Chebouti1and F/Zohra Bissaad

Abstract:

Essential oils have a significant antimicrobial activity. These oils can successfully replace the antibiotics. So, the microorganisms show their inefficiencies resistant for the antibiotics. For this reason, we study the physicochemical analysis and antimicrobial activity of the essential oil of Daucus carota. The extraction is done by steam distillation of water which brought us a very significant return of 4.65%. The analysis of the essential oil is performed by GC/MS and has allowed us to identify 32 compounds in the oil of D. carota flowering tops of Bouira. Three of which are in the majority are the α-pinene (22.3%), the carotol (21.7%) and the limonene (15.8%).

Keywords: Daucus carota, essential oil, α-pinene, carotol, limonene

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Authors: Shubham Sharma, Swarna Jaiswal, Brendan Duffy, Amit Jaiswal

Abstract:

The key features of any active packaging film are its biodegradability and antimicrobial properties. Biological macromolecules such as polyphenols (ferulic acid (FA) and tannic acids (TA)) are naturally found in plants such as grapes, berries, and tea. In this study, antimicrobial activity screening of several polyphenols was carried out by using minimal inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against two strains of gram-negative bacteria - Salmonella typhimurium, Escherichia coli, and two-gram positive strains - Staphylococcus aureus and Listeria monocytogenes. FA and TA had shown strong antibacterial activity at the low concentration against both gram-positive and gram-negative bacteria. The selected polyphenols FA and TA were incorporated at various concentrations (1%, 5%, and 10% w/w) in the poly(lactide) – poly (butylene adipate-co-terephthalate) (PLA-PBAT) composite film by using the solvent casting method. The effect of TA and FA incorporation in the packaging was characterized based on morphological, optical, color, mechanical, thermal, and antimicrobial properties. The thickness of the FA composite film was increased by 1.5 – 7.2%, while for TA composite film, it increased by 0.018 – 1.6%. FA and TA (10 wt%) composite film had shown approximately 65% - 66% increase in the UV barrier property. As the FA and TA concentration increases from 1% - 10% (w/w), the TS value increases by 1.98 and 1.80 times, respectively. The water contact angle of the film was observed to decrease significantly with the increase in the FA and TA content in the composite film. FA has shown more significant increase in antimicrobial activity than TA in the composite film against Listeria monocytogenes and E. coli. The FA and TA composite film has the potential for its application as an active food packaging.

Keywords: active packaging, biodegradable film, polyphenols, UV barrier, tensile strength

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Authors: Suliman Khan, Rabeea Siddique, Mengzhou Xue

Abstract:

Background: Patients with neurological disorders often display altered circadian rhythms. The disrupted circadian rhythms through chronic jetlag or shiftwork are thought to increase the risk and severity of human disease, including cancer, psychiatric, and related brain diseases. In this study, we investigated the impact of shiftwork or chronic jetlag (CJL) like conditions on mice’s brains. Transcriptome profiling based on RNA sequencing revealed that genes associated with serious neurological disorders were differentially expressed in the nucleus accumbens (NAc) and prefrontal cortex (PFC). According to the qPCR analysis, several key regulatory genes associated with neurological disorders were significantly altered in the NAc, PFC, hypothalamus, hippocampus, and striatum. Serotonin levels and the expression levels of serotonin transporters and receptors were significantly altered in mice treated with CJL. Overall, these results indicate that CJL may increase the risk of neurological disorders by disrupting the key regulatory genes, biological functions, serotonin, and corticosterone. These molecular linkages can further be studied to investigate the mechanism underlying CJL or shiftwork-mediated neurological disorders in order to develop treatment strategies.

Keywords: chronic jetlag, molecular profiles, brain disorders, circadian rhythms

Procedia PDF Downloads 97