Identifying potential RNAi targets in aphid species (Macrosiphum rosae, Rhopalosiphum maidis, Sitobion avenae, Toxoptera aurantii) in sub-tropical region with extreme summer

Abstract RNA interference is a useful and efficient tool that had been used to incorporate tolerance against different stresses. Five unigene sequences were selected from exotic grain aphid that were reported to be an ideal RNAi targets. Aphids (Macrosiphum rosae, Rhopalosiphum maidis, Sitobion avenae, Toxoptera aurantii) were reared on Rosa indica, Zea mays, Triticum aestivum, Chrysanthemum hibiscus, Solanum melongena, Abelmoschus esculentus. PCR of unigene 28469 and 21789 was positive for all aphids and there is no ortholog found for them so unique and effective to be used in RNAi technology. Cytochrome c oxidase was found to be positive for M. rosae, S. avenae, T. aurantii and negative for R. maidis. Zinc finger protein was found to be positive for R. maidis, S. avenae and negative for M. rosae, and T. aurantii. Cuticular proteins was found to be positive for S. avenae and M. rosae and negative for R. maidis and T. aurantii. Genes identified in the aphids are defense and as important structural genes and their suppression with RNAi technology will be important target to have insect resistant crops. From these gene sequences cytochrome c oxidase is reported as bar coding gene and can be used in future for interspecific genetic variation in these aphids. Results revealed the high expression of these sequences in local aphid species and can be used as RNAi target for them. That can be used in future or applications as pest management, monitoring and plant quarantine

Cardiff Acne Disability Index based Quality of Life in Acne Patients, Risk Factors and Associations

BACKGROUND: Acne Vulgaris is a frequently seen dermatological condition having physical and psychosocial implications. Current study aims to assess Quality of life (QoL) in acne patients and study the associated risk factors. METHODOLOGY: This descriptive cross sectional study was conducted in 6 months duration at Dept. of Dermatology, Rawal Institute of Health Sciences Islamabad after ethical approval. Total 205 cases of acne (age>12 years) were included. Patients with severe systemic illness and endocrine disease were excluded. Global Acne Grading Scale (GAGS) was used to grade acne as mild, moderate or severe. Quality of life assessed by Cardiff Acne disability index (CADI) with outcome as no impairment, mild impairment, moderate impairment or severe impairment. Data analyzed via SPSS version 17 with Chi-square test applied for significance. The p-value <0.05 considered to be statistically significant. RESULTS: Among 205 acne cases (24% males and 76% females), mean age was 25+5.7 (14-36) years. As per GAGS score; mild acne was present in 45.4%, moderate acne(44%), severe acne (8.35%) and very severe acne (2.4%). As per CADI score, there was no impairment in 7.3%, mild mpairment (19.5%), moderate impairment (61%) and severe impairment in (12%). The impaired QoL was found to be associated with gender, socioeconomic class, education, acne grade, obesity and use of alternative medicine (p<0.05). CONCLUSION: Acne vulgaris significantly impairs QoL in adolescents to varying degrees. Higher grade of acne, female gender, obesity, illiteracy and poverty are the proposed risk factors for impaired QoL in acne cases. Early identification of acne cases at risk of impaired Qol may be helpful to take QoL measures like counseling and psychological intervention in parallel with medical management. This may improve the social and psychological wellbeing of acne cases

Surveillance of molecular markers of antimalarial drug resistance in Plasmodium falciparum and Plasmodium vivax in Federally Administered Tribal Area (FATA), Pakistan

This molecular epidemiological study was designed to determine the antimalarial drug resistance pattern, and the genetic diversity of malaria isolates collected from a war-altered Federally Administered Tribal Area (FATA), in Pakistan. Clinical isolates were collected from Bajaur, Mohmand, Khyber, Orakzai and Kurram agencies of FATA region between May 2017 and May 2018, and they underwent DNA extraction and amplification. The investigation of gene polymorphisms in drug resistance genes (dhfr, dhps, crt, and mdr1) of Plasmodium falciparum and Plasmodium vivax was carried out by pyrosequencing and Sanger sequencing, respectively. Out of 679 PCR-confirmed malaria samples, 523 (77%) were P. vivax, 121 (18%) P. falciparum, and 35 (5%) had mixed-species infections. All P. falciparum isolates had pfdhfr double mutants (C59R+S108N), while pfdhfr/pfdhps triple mutants (C59R+S108N+A437G) were detected in 11.5% of the samples. About 97.4% of P. falciparum isolates contained pfcrt K76T mutation, while pfmdr1 N86Y and Y184F mutations were present in 18.2% and 10.2% of the samples. P. vivax pvdhfr S58R mutation was present in 24.9% of isolates and the S117N mutation in 36.2%, while no mutation in the pvdhps gene was found. Pvmdr1 F1076L mutation was found in nearly all samples, as it was observed in 98.9% of isolates. No significant anti-folate and chloroquine resistance was observed in P. vivax; however, mutations associated with antifolate-resistance were found, and the chloroquine-resistant gene has been observed in 100% of P. falciparum isolates. Chloroquine and sulphadoxine-pyrimethamine resistance were found to be high in P. falciparum and low in P. vivax. Chloroquine could still be used for P. vivax infection but need to be tested in vivo, whereas a replacement of the artemisinin combination therapy for P. falciparum appears to be justified

Diversity Assessment and Cultivar Identification in Date Palm through Molecular Markers- A Review

Date palm has a long history of cultivation and a valuable germplasm with little knowledge about its genetic makeup and variation among the most cultivated cultivars. Diversity is the variability of a species. Plants show variation in yield, vegetative traits and morphological properties of fruits and seeds in response to environmental changes. Molecular markers or DNA markers have been in use since past three decades. The DNA profiles give information about the genotype, screen the whole genome and show variation in both the coding and noncoding region and hence give information about polymorphism. Since plastid genes are transferred mostly from the mother line, the identification of maternal lines is possible by the sequencing of plastid genes. Simple sequence repeats (SSRs) can detect length variation with the help of Polymerase Chain Reaction (PCR) and may be used as highly informative genetic markers. Single Nucleotide Polymorphism (SNPs) are the third generation of molecular markers. SNPs are more stable and have high fidelity of inheritance as compared to other marker systems. Molecular markers have been developed but they are not enough for sufficient diversity assessment. Therefore there is a need to increase the number of DNA markers in date palm. Previously, there are several studies to type various commercially important germplasm based on morphological or yield parameters. Morphological and biochemical markers are limited in number and are affected by environmental factors and growth stage of the plant which reduce their reliability in the assessment of diversity and characterization of the germplasm. This necessitates the use of genetic characterization, utilizing DNA markers, gene sequencing or SNP genotyping which can work independent of the plant growth stage and are not affected by environmental factors. A combination of morphological, biochemical and molecular characterization of the date palm cultivars can better assess the level of diversity and relationship among the cultivars

Heterologous expression and enhanced production of β-1,4-glucanase of Bacillus halodurans C-125 in Escherichia coli

Background: Recombinant DNA technology enables us to produce proteins with desired properties and insubstantial amount for industrial applications. Endo-1, 4-β-glucanases (Egl) is one of the major enzyme involved in degradation of cellulose, an important component of plant cell wall. The present study was aimed at enhancing the production of endo-1, 4-β-glucanases (Egl) of Bacillus halodurans in Escherichia coli. Results: A putative Egl gene of Bacillus Halodurans was expressed in E. coli by cloning in pET 22b (+). On induction with isopropyl-b-d-1-thiogalactopyranoside, the enzyme expression reached upto ~20% of the cell protein producing 29.2 mg/liter culture. An increase in cell density to 12 in auto-inducing LB medium (absorbance at 600 nm) enhanced β-glucanase production up to 5.4 fold. The molecular mass of the enzyme was determined to be 39 KDa, which is nearly the same as the calculated value. Protein sequence was analyzed by CDD, Pfam, I TASSER, COACH, PROCHECK Servers and putative amino acids involved in the formation of catalytic, substrate and metal binding domains were identified. Phylogenetic analysis of the β-glucanases of B. halodurans was performed and position of Egl among other members of the genus Bacillus producing endo-glucanases was determined. Temperature and pH optima of the enzyme were found to be 60°C and 8.0, respectively, under the assay conditions. Conclusion: Production of endo-1, 4 β-glucanase enzymes from B. halodurans increased several folds when cloned in pET vector and expressed in E. coli. To our knowledge, this is the first report of high-level expression and characterization of an endo-1, 4 β-glucanases from B. halodurans.How to cite: Zeeshan N, Naz S, Naz S et al. Heterologous expression and enhanced production of β-1,4-glucanase of Bacillus halodurans C-125 in E. coli. Electron J Biotechnol 2018;34. https://doi.org/10.1016/j.ejbt.2018.05.001. Keywords: Bacillus halodurans, Cellulases, Cellulose hydrolysis, Degradation of cellulose, Endo-1, 4-β-glucanases, Expression analysis, Heterologous expression, In silico protein characterization, IPTG, pET expression system, Plant cell wal

Management of Hinchey II diverticulitis.

Colonic diverticulosis can either be asymptomatic or present with complications resulting in significant morbidity and mortality. A key presentation of complicated disease is abscess formation (Hinchey type II). The natural course of this is unclear and therefore treatments range from conservative approach with antibiotics and percutaneous guided drainage (PCD) to surgery. There is no clear consensus on the exact management strategy. A Medline based literature search specifically looking at studies dealing with Hinchey type II diverticulitis and its management was carried out. For comparison, five-year retrospective data of diverticular abscesses from our institution was collected and the outcome analysed. Various studies have looked into this aspect of the disease, elaborating on the significance of the size and location of the abscesses, the role of PCD, recurrence rates and the controversies regarding the need for elective surgery. Conservative treatment with antibiotics alone is effective in a majority of cases with a role for PCD in large safely accessible abscesses. Variable recurrence rates have been reported in literature and elective surgery should be planned for selected groups of patients

Immobilization of fungal cellulase on calcium alginate and xerogel matrix

We conducted a study for immobilization of cellulase produced by Aspergillus tubingensis-IMMIS2 through pre-optimized solid-state fermentation of corn stover. Incredible increment in stability and catalytic activity of the immobilized cellulase was observed. Thermostability of the immobilized cellulase was increased to 82% at 75\ua0°C as compared to that of free cellulase after 26\ua0h of incubation. The cellulase activity was decreased after the 20th day of incubation of the both immobilized and free enzymes. Maximum cellulase activity was achieved at pH 4.5 (174 ± 0.4 U mL min ) and temperature 45\ua0°C (179 ± 0.4 U mL min ) for xerogel matrix. The lowest K value was found for the enzyme immobilized on xerogel as compared to those of\ua0immobilized on calcium alginate and free enzyme. Immobilization of cellulase on calcium alginate and xerogel matrix increased tolerance capacity of the enzyme to 75–82% against different activators and/or inhibitors like EDTA, SDS, Co , Ca and Hg . The immobilized cellulase also revealed good fruit saccharification and yielded increased juice volume and hence proved to be a suitable candidate for biotechnological and industrial applications

International Students’ Nostalgic Behaviour towards the Purchase of Products and Services

The literature has highlighted that international students abroad exhibit a strong preference for products from their country compared with foreign products and services, thus suggesting that the cultural disposition of consumers influences their purchasing decisions. Therefore, this current research aimed to determine how the feeling of groundedness could affect international students’ purchasing behavior in Malaysia through cultural products and services. To this end, quantitative data were collected from international students at the University of Malaya and the University Utara Malaysia and then analyzed using a structural equation model (SEM). The findings of this research highlighted that consumer behavior was influenced by a “feeling of groundedness”. This suggested that cultural services and products are valued by international students in foreign universities because of their ability to evoke a feeling of groundedness. The theoretical contributions of this study, particularly to the consumer behavior literature, are extensive, including how it highlights the influence of the feeling of groundedness on international consumers’ purchasing behavior. The marketing implications of this research are also valuable for businesses targeting international students