Cloning, sequence analysis, and expression of cDNA coding for the major house dust mite allergen, Der f 1, in Escherichia coli

Our objective was to clone, express and characterize adult Dermatophagoides farinae group 1 (Der f 1) allergens to further produce recombinant allergens for future clinical applications in order to eliminate side reactions from crude extracts of mites. Based on GenBank data, we designed primers and amplified the cDNA fragment coding for Der f 1 by nested-PCR. After purification and recovery, the cDNA fragment was cloned into the pMD19-T vector. The fragment was then sequenced, subcloned into the plasmid pET28a(+), expressed in Escherichia coli BL21 and identified by Western blotting. The cDNA coding for Der f 1 was cloned, sequenced and expressed successfully. Sequence analysis showed the presence of an open reading frame containing 966 bp that encodes a protein of 321 amino acids. Interestingly, homology analysis showed that the Der p 1 shared more than 87% identity in amino acid sequence with Eur m 1 but only 80% with Der f 1. Furthermore, phylogenetic analyses suggested that D. pteronyssinus was evolutionarily closer to Euroglyphus maynei than to D. farinae, even though D. pteronyssinus and D. farinae belong to the same Dermatophagoides genus. A total of three cysteine peptidase active sites were found in the predicted amino acid sequence, including 127-138 (QGGCGSCWAFSG), 267-277 (NYHAVNIVGYG) and 284-303 (YWIVRNSWDTTWGDSGYGYF). Moreover, secondary structure analysis revealed that Der f 1 contained an a helix (33.96%), an extended strand (17.13%), a ß turn (5.61%), and a random coil (43.30%). A simple three-dimensional model of this protein was constructed using a Swiss-model server. The cDNA coding for Der f 1 was cloned, sequenced and expressed successfully. Alignment and phylogenetic analysis suggests that D. pteronyssinus is evolutionarily more similar to E. maynei than to D. farinae.

Cloning, bioinformatics analysis, and expression of the dust mite allergen Der f 5 of Dermatophagoides farinae

Crude extracts of house dust mites are used clinically for diagnosis and immunotherapy of allergic diseases, including bronchial asthma, perennial rhinitis, and atopic dermatitis. However, crude extracts are complexes with non-allergenic antigens and lack effective concentrations of important allergens, resulting in several side effects. Dermatophagoides farinae (Hughes; Acari: Pyroglyphidae) is one of the predominant sources of dust mite allergens, which has more than 30 groups of allergen. The cDNA coding for the group 5 allergen of D. farinae from China was cloned, sequenced and expressed. According to alignment using the VECTOR NTI 9.0 software, there were eight mismatched nucleotides in five cDNA clones resulting in seven incompatible amino acid residues, suggesting that the Der f 5 allergen might have sequence polymorphism. Bioinformatics analysis revealed that the matured Der f 5 allergen has a molecular mass of 13604.03 Da, a theoretical pI of 5.43 and is probably hydrophobic and cytoplasmic. Similarities in amino acid sequences between Der f 5 and allergens of other domestic mite species, viz. Der p 5, Blo t 5, Sui m 5, and Lep d 5, were 79, 48, 53, and 37%, respectively. Phylogenetic analysis indicated that Der f 5 and Der p 5 clustered together. Blo t 5 and Ale o 5 also clustered together, although Blomia tropicalis and Aleuroglyphus ovatus belong to different mite families, viz. Echimyopodidae and Acaridae, respectively.

Invoice process analysis and development. Case: Power and automation technology company

This master’s thesis studies the case company’s current purchase invoice process and the challenges that are related to it. Like most of other master’s thesis this study consists of both theoretical- and empirical parts. The purpose of this work is to combine theoretical and empirical parts together so that the theoretical part brings value to the empirical case study. The case company’s main business is frequency converters for both low voltage AC & DC drives and medium voltage AC Drives which are used across all industries and applications. The main focus of this study is on the current invoice process modelling. When modelling the existing process with discipline and care, current challenges can be understood better. Empirical study relays heavily on interviews and existing, yet fragmented, data. This, along with own calculations and analysis, creates the foundation for the empirical part of this master’s thesis.

Physiochemical analysis and centesimal composition of Pleurotus ostreatus mushroom grown in residues from the Amazon

The objective of the present work was to evaluate the nutritional composition of mushrooms produced in alternative substrates in agricultural and agro-industrial residues from the Amazon. C, N, pH, moisture, soluble solids, protein, lipids, total fibers, ashes, carbohydrates and energy were determined. Substrates were formulated from Simarouba amara Aubl. and Ochroma piramidale Cav. ex. Lam. Sawdust and from Bactris gasipaes Kunth and Saccharum officinarum stipe. Results showed that the nutritional composition of P. ostreatus varied according to the cultivation substrate and that it can be considered important food due to its nutritional characteristics such as: high protein content; metabolizable carbohydrates and fiber; and low lipids and calories contents.

Evaluation of antioxidants stability by thermal analysis and its protective effect in heated edible vegetable oil

In this work, through the use of thermal analysis techniques, the thermal stabilities of some antioxidants were investigated, in order to evaluate their resistance to thermal oxidation in oils, by heating canola vegetable oil, and to suggest that antioxidants would be more appropriate to increase the resistance of vegetable oils in the thermal degradation process in frying. The techniques used were: Thermal Gravimetric (TG) and Differential Scanning Calorimetry (DSC) analyses, as well as an allusion to a possible protective action of the vegetable oils, based on the thermal oxidation of canola vegetable oil in the laboratory under constant heating at 180 ºC/8 hours for 10 days. The studied antioxidants were: ascorbic acid, sorbic acid, citric acid, sodium erythorbate, BHT (3,5-di-tert-butyl-4-hydroxytoluene), BHA (2, 3-tert-butyl-4-methoxyphenol), TBHQ (tertiary butyl hydroquinone), PG (propyl gallate) - described as antioxidants by ANVISA and the FDA; and also the phytic acid antioxidant and the SAIB (sucrose acetate isobutyrate) additive, which is used in the food industry, in order to test its behavior as an antioxidant in vegetable oil. The following antioxidants: citric acid, sodium erythorbate, BHA, BHT, TBHQ and sorbic acid decompose at temperatures below 180 ºC, and therefore, have little protective action in vegetable oils undergoing frying processes. The antioxidants below: phytic acid, ascorbic acid and PG, are the most resistant and begin their decomposition processes at temperatures between 180 and 200 ºC. The thermal analytical techniques have also shown that the SAIB antioxidant is the most resistant to oxidative action, and it can be a useful choice in the thermal decomposition prevention of edible oils, improving stability regarding oxidative processes.

Hygienic-sanitary working practices and implementation of a Hazard Analysis and Critical Control Point (HACCP) plan in lobster processing industries

This study aimed to verify the hygienic-sanitary working practices and to create and implement a Hazard Analysis Critical Control Point (HACCP) in two lobster processing industries in Pernambuco State, Brazil. The industries studied process frozen whole lobsters, frozen whole cooked lobsters, and frozen lobster tails for exportation. The application of the hygienic-sanitary checklist in the industries analyzed achieved conformity rates over 96% to the aspects evaluated. The use of the Hazard Analysis Critical Control Point (HACCP) plan resulted in the detection of two critical control points (CCPs) including the receiving and classification steps in the processing of frozen lobster and frozen lobster tails, and an additional critical control point (CCP) was detected during the cooking step of processing of the whole frozen cooked lobster. The proper implementation of the Hazard Analysis Critical Control Point (HACCP) plan in the lobster processing industries studied proved to be the safest and most cost-effective method to monitor each critical control point (CCP) hazards.

A comparative study among methods used for wheat flour analysis and for measurements of gluten properties using the Wheat Gluten Quality Analyser (WGQA)

This study aimed at comparing both the results of wheat flour quality assessed by the new equipment Wheat Gluten Quality Analyser (WGQA) and those obtained by the extensigraph and farinograph. Fifty-nine wheat samples were evaluated for protein and gluten contents; the rheological properties of gluten and wheat flour were assessed using the WGQA and the extensigraph/farinograph methods, respectively, in addition to the baking test. Principal component analysis (PCA) and linear regression were used to evaluate the results. The parameters of energy and maximum resistance to extension determined by the extensigraph and WGQA showed an acceptable level for the linear correlation within the range from 0.6071 to 0.6511. The PCA results obtained using WGQA and the other rheological apparatus showed values similar to those expected for wheat flours in the baking test. Although all equipment used was effective in assessing the behavior of strong and weak flours, the results of medium strength wheat flour varied. WGQA has shown to use less amount of sample and to be faster and easier to use in relation to the other instruments used.

Power Electronic Converters in Low-Voltage Direct Current Distribution – Analysis and Implementation

Over the recent years, smart grids have received great public attention. Many proposed functionalities rely on power electronics, which play a key role in the smart grid, together with the communication network. However, “smartness” is not the driver that alone motivates the research towards distribution networks based on power electronics; the network vulnerability to natural hazards has resulted in tightening requirements for the supply security, set both by electricity end-users and authorities. Because of the favorable price development and advancements in the field, direct current (DC) distribution has become an attractive alternative for distribution networks. In this doctoral dissertation, power electronic converters for a low-voltage DC (LVDC) distribution system are investigated. These include the rectifier located at the beginning of the LVDC network and the customer-end inverter (CEI) on the customer premises. Rectifier topologies are introduced, and according to the LVDC system requirements, topologies are chosen for the analysis. Similarly, suitable CEI topologies are addressed and selected for study. Application of power electronics into electricity distribution poses some new challenges. Because the electricity end-user is supplied with the CEI, it is responsible for the end-user voltage quality, but it also has to be able to supply adequate current in all operating conditions, including a short-circuit, to ensure the electrical safety. Supplying short-circuit current with power electronics requires additional measures, and therefore, the short-circuit behavior is described and methods to overcome the high-current supply to the fault are proposed. Power electronic converters also produce common-mode (CM) and radio-frequency (RF) electromagnetic interferences (EMI), which are not present in AC distribution. Hence, their magnitudes are investigated. To enable comprehensive research on the LVDC distribution field, a research site was built into a public low-voltage distribution network. The implementation was a joint task by the LVDC research team of Lappeenranta University of Technology and a power company Suur-Savon S¨ahk¨o Oy. Now, the measurements could be conducted in an actual environment. This is important especially for the EMI studies. The main results of the work concern the short-circuit operation of the CEI and the EMI issues. The applicability of the power electronic converters to electricity distribution is demonstrated, and suggestions for future research are proposed.

Analysis and guidelines for implementation of EN 1090-2 Standard to assist execution class assignment of steel structures

Finnish design and consulting companies are delivering robust and cost-efficient steel structures solutions to a large number of manufacturing companies worldwide. Recently introduced EN 1090-2 standard obliges these companies to specify the execution class of steel structures for their customers. This however, requires clarifying, understanding and interpreting the sophisticated procedure of execution class assignment. The objective of this research is to provide a clear explanation and guidance through the process of execution class assignment for a given steel structure and to support the implementation of EN 1090-2 standard in Rejlers Oy, one of Finnish design and consulting companies. This objective is accomplished by creating a guideline for designers that elaborates on the four-step process of the execution class assignment for a steel structure or its part. Steps one to three define the consequence class (projected consequences of structure failure), the service category (hazards associated with the service use exploitation of steel structure) and the production category (manufacturing process peculiarities), based on the ductility class (capacity of structure to withstand deformations) and the behaviour factor (corresponds to structure seismic behaviour). The final step is the execution class assignment taking into account results of previous steps. Main research method is indepth literature review of European standards family for steel structures. Other research approach is a series of interviews of Rejlers Oy representatives and its clients, results of which have been used to evaluate the level of EN 1090-2 awareness. Rejlers Oy will use the developed novel coherent standard implementation guideline to improve its services and to obtain greater customer satisfaction.

Social-business tensions and their management in social enterprise: evidence from Finland.

This thesis aims to provide insight into the social-business tensions the social enterprises face in their operation and how they manage them. The social-business tensions are examined from four theoretical perspectives using triangulation approach. The theoretical lenses chosen are organizational identity, stakeholder theory, paradox theory and institutional theory. The theories aim to clarify, how the tensions are formed, how they appear and how they are managed in social enterprises. One viewpoint of this thesis is to examine the competence of these theories in explaining the social-business tensions in practise. The qualitative data was collected by interviewing persons from the management of two social enterprises. The empirical evidence of this thesis suggests that the appearing of social-business tensions varies between the social enterprises and they can be seen both as an advantage and as a challenge. Most of the social-business tensions arise from the enterprise’s multiple incoherent objectives, their stakeholders’ various demands and the differing understanding of the company’s central operation among the members of the organization. According to this thesis, the theories of organizational identity, stakeholder, paradox and institution are all able to provide unique insight into the identification and management of the social-business tensions. However, the paradox theory turned out to be the most abstract of the theories and thus being the farthest from the practise.

Spurring innovation through idea contests : data analysis and process improvements

This research concerns the Urban Living Idea Contest conducted by Creator Space™ of BASF SE during its 150th anniversary in 2015. The main objectives of the thesis are to provide a comprehensive analysis of the Urban Living Idea Contest (ULIC) and propose a number of improvement suggestions for future years. More than 4,000 data points were collected and analyzed to investigate the functionality of different elements of the contest. Furthermore, a set of improvement suggestions were proposed to BASF SE. Novelty of this thesis lies in the data collection and the original analysis of the contest, which identified its critical elements, as well as the areas that could be improved. The author of this research was a member of the organizing team and involved in the decision making process from the beginning until the end of the ULIC.