Production and characterization of fungal b-glucosidase and bacterial cellulases by tobacco chloroplast transformation

Edward Alexander Espinoza-Sanchez,Jorge Ariel Torres-Castillo,Quintın Rascon-Cruz,Francisco Zavala-Garcıa,Sugey Ramona Sinagawa-Garcıa 한국식물생명공학회 2016 Plant biotechnology reports Vol.10 No.2

The high capacity of the chloroplast genome to integrate and express transgenes at high levels makes transplastomic technology a good option for overexpressing proteins of interest. This report presents the stable expression of b-glucosidase (bgl1 gene) from Aspergillus niger and two cellulases (celA and celB genes) from Thermotoga neapolitana into the chloroplast genome of tobacco. The pES6, pHM4, pHM5 and pHM6 vectors were derived from the pES4 plasmid containing bgl1, celA-celB, celA and celB synthetic genes, respectively. All of the genes were flanked by a synthetic rrn16 promoter and the 30UTR from rbcL gene. The integration of the genes into intergenic regions rrn16 and 30rps12 of the inverted repeats was confirmed by Southern blot analysis. Stable expression and processing of monocistronic mRNA were confirmed by Northern blot analysis, and protein functionality was analysed via enzymatic activity assay. The recombinant enzymes exhibited high enzymatic activity at pH 5 (bglucosidase: 30.45 U mg-1 of TSP, celA-celB 58 U mg-1 of TSP, celA 49.10 U mg-1 of TSP and celB 48.72 U mg-1 of TSP). In addition, b-glucosidase exhibited high activity at 40 ℃, whereas cellulases type A (celA) and type B (celB) showed high activity at 65 ℃. NtpES6, NtpHM5 and NtpHM6 plants showed a similar phenotype compared with the wild type plants; however, NtpHM4 plants presented an abnormal phenotype with variegated leaves. This study, demonstrated that hydrolytic genes such as bgl1, celA and celB could be integrated and expressed correctly in the chloroplast genome. This work provides new information on methods and strategies for the expression of hydrolytic enzymes that are potentially useful for biotechnological applications using transplastomic plants.

Bloodstream Infections and Frequency of Pretreatment Associated With Age and Hospitalization Status in Sub-Saharan Africa

Nichols, Chelsea,Cruz Espinoza, Ligia Maria,von Kalckreuth, Vera,Aaby, Peter,Ahmed El Tayeb, Muna,Ali, Mohammad,Aseffa, Abraham,Bjerregaard-Andersen, Morten,Breiman, Robert F.,Cosmas, Leonard,Crump, J Oxford University Press 2015 Clinical infectious diseases Vol.61 No.suppl4

<P><B><I>Background.</I></B> The clinical diagnosis of bacterial bloodstream infections (BSIs) in sub-Saharan Africa is routinely confused with malaria due to overlapping symptoms. The Typhoid Surveillance in Africa Program (TSAP) recruited febrile inpatients and outpatients of all ages using identical study procedures and enrollment criteria, thus providing an opportunity to assess disease etiology and pretreatment patterns among children and adults.</P><P><B><I>Methods.</I></B> Inpatients and outpatients of all ages with tympanic or axillary temperatures of ≥38.0 or ≥37.5°C, respectively, and inpatients only reporting fever within the previous 72 hours were eligible for recruitment. All recruited patients had one blood sample drawn and cultured for microorganisms. Data from 11 TSAP surveillance sites in nine different countries were used in the analysis. Bivariate analysis was used to compare frequencies of pretreatment and BSIs in febrile children (<15 years old) and adults (≥15 years old) in each country. Pooled Cochran Mantel–Haenszel odds ratios (ORs) were calculated for overall trends.</P><P><B><I>Results.</I></B> There was no significant difference in the odds of a culture-proven BSI between children and adults among inpatients or outpatients. Among both inpatients and outpatients, children had significantly higher odds of having a contaminated blood culture compared with adults. Using country-pooled data, child outpatients had 66% higher odds of having <I>Salmonella</I> Typhi in their bloodstream than adults (OR, 1.66; 95% confidence interval [CI], 1.01–2.73). Overall, inpatient children had 59% higher odds of pretreatment with analgesics in comparison to inpatient adults (OR, 1.59; 95% CI, 1.28–1.97).</P><P><B><I>Conclusions.</I></B> The proportion of patients with culture-proven BSIs in children compared with adults was similar across the TSAP study population; however, outpatient children were more likely to have <I>Salmonella</I> Typhi infections than outpatient adults. This finding points to the importance of including outpatient facilities in surveillance efforts, particularly for the surveillance of typhoid fever. Strategies to reduce contamination among pediatric blood cultures are needed across the continent to prevent the misdiagnosis of BSI cases in children.</P>

What is a traditional food? Conceptual evolution from four dimensions

Rocillo-Aquino Zeltzin,Cervantes-Escoto Fernando,Leos-Rodríguez Juan Antonio,Cruz-Delgado Daniela,Espinoza-Ortega Angélica 한국식품연구원 2021 Journal of Ethnic Foods Vol.8 No.-

The purpose was to specify the concept of traditional food and the dimensions that make it up, identifying: definitions, authors and research projects; as well as to determine what is known and possible topics for future research. A literature review of traditional foods was conducted that examined the conceptual development of the term. Social network analysis (SNA) was also used to identify the most relevant definitions and working groups on the topic. Twenty-three definitions were identified in the period 1995–2019. It reveals the difficulty of establishing one that ecompasses such a dynamic concept. Although there is variability in the specific characteristics of these foods, four dimensions have been established: time, place, know-how, and cultural meaning. It was found that their main characteristic is the transmission of knowledge and raw materials between generations. The conceptualization of the term has been developed mainly in Europe, based on the perspective of consumers. New trends in research include the contrast and complementarity of innovation in traditional foods and the difference between these products and those named with similar attributes such as typical, regional, ethnic, local, among others. Social network analysis (SNA) was used to study the definitions of a concept, something that had not been done with this approach. Suggestions are made for possible research on the subject, such as the conceptual delimitation of related terms and the compatibility between innovation and tradition. The purpose was to specify the concept of traditional food and the dimensions that make it up, identifying: definitions, authors and research projects; as well as to determine what is known and possible topics for future research. A literature review of traditional foods was conducted that examined the conceptual development of the term. Social network analysis (SNA) was also used to identify the most relevant definitions and working groups on the topic. Twenty-three definitions were identified in the period 1995–2019. It reveals the difficulty of establishing one that encompasses such a dynamic concept. Although there is variability in the specific characteristics of these foods, four dimensions have been established: time, place, know-how, and cultural meaning. It was found that their main characteristic is the transmission of knowledge and raw materials between generations. The conceptualization of the term has been developed mainly in Europe, based on the perspective of consumers. New trends in research include the contrast and complementarity of innovation in traditional foods and the difference between these products and those named with similar attributes such as typical, regional, ethnic, local, among others. Social network analysis (SNA) was used to study the definitions of a concept, something that had not been done with this approach. Suggestions are made for possible research on the subject, such as the conceptual delimitation of related terms and the compatibility between innovation and tradition.