Viral membrane fusion: is glycoprotein G of rhabdoviruses a representative of a new class of viral fusion proteins?

Enveloped viruses always gain entry into the cytoplasm by fusion of their lipid envelope with a cell membrane. Some enveloped viruses fuse directly with the host cell plasma membrane after virus binding to the cell receptor. Other enveloped viruses enter the cells by the endocytic pathway, and fusion depends on the acidification of the endosomal compartment. In both cases, virus-induced membrane fusion is triggered by conformational changes in viral envelope glycoproteins. Two different classes of viral fusion proteins have been described on the basis of their molecular architecture. Several structural data permitted the elucidation of the mechanisms of membrane fusion mediated by class I and class II fusion proteins. In this article, we review a number of results obtained by our laboratory and by others that suggest that the mechanisms involved in rhabdovirus fusion are different from those used by the two well-studied classes of viral glycoproteins. We focus our discussion on the electrostatic nature of virus binding and interaction with membranes, especially through phosphatidylserine, and on the reversibility of the conformational changes of the rhabdovirus glycoprotein involved in fusion. Taken together, these data suggest the existence of a third class of fusion proteins and support the idea that new insights should emerge from studies of membrane fusion mediated by the G protein of rhabdoviruses. In particular, the elucidation of the three-dimensional structure of the G protein or even of the fusion peptide at different pH's might provide valuable information for understanding the fusion mechanism of this new class of fusion proteins.

Human T-cell lymphotropic virus type I (HTLV-I) proviral DNA viral load among asymptomatic patients and patients with HTLV-I-associated myelopathy/tropical spastic paraparesis

To evaluate the human T-cell lymphotropic virus type I (HTLV-I) proviral DNA load among asymptomatic HTLV-I-infected carriers and patients with HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP), real time PCR using TaqMan probes for the pol gene was performed in two million peripheral blood mononuclear cells (PBMC). The albumin gene was the internal genomic control and MT2 cells were used as positive control. The results are reported as copies/10,000 PBMC, and the detection limit was 10 copies. A total of 89 subjects (44 HAM/TSP and 45 healthy HTLV-I-infected carriers) followed up at the Institute of Infectious Diseases "Emilio Ribas" and in the Neurology Division of Hospital of Clínicas were studied. The asymptomatic HTLV-I-infected carriers had a median number of 271 copies (ranging from 5 to 4756 copies), whereas the HAM/TSP cases presented a median of 679 copies (5-5360 copies) in 10,000 PBMC. Thus, HAM/TSP patients presented a significantly higher HTLV-I proviral DNA load than healthy HTLV-I carriers (P = 0.005, one-way Mann-Whitney test). As observed in other persistent infections, proviral DNA load quantification may be an important tool for monotoring HTLV-I-infected subjects. However, long-term follow-up is necessary to validate this assay in the clinical setting.

MARKETING COMMUNICATIONS IN B2B COMPANIES – The Roles of Marketing Communication Tools in the Digital Age

Marketing communications has gone through significant changes during the last decades and new online tools have been leading this change for the last decade. Now, in the digital age, if wanting to be successful companies need to experiment new things and seize the opportunities online provides and adapt to the new environment. However, during this time the marketing communication mix and the meanings given to the various components of it have not changed dramatically, and personal selling, direct marketing, sales promotion, advertising and public relations activities are still regarded as important tools in the marketing communications mix. The purpose of this study was to examine business-to-business marketing communications and the tools used by companies in their marketing communications efforts in the digital age and in a global environment. The research questions dealt with the marketing communication tools and their roles as well as the role of online marketing communication and the way it has shaped the field of b2b marketing communications. In order to answer these questions, qualitative approach was chosen and the data was collected by theme interviews with six representatives of Finnish global companies from the b2b sector being interviewed. The theoretical framework covers the general field of b2b marketing communications and its main elements. The online environment as well as the concept of a global marketplace and integrated approach to marketing communication activities is also discussed. The theory was supported by the interviews regarding the activities and roles of the marketing communication tools and both theory as well as the interviews found personal selling to be a vital tool. However, the importance of online has grown and online marketing activities have risen right next to personal selling. The use of analytics and marketing automation was found to be of great interest in the interviews and they were seen as a growing domain in b2b marketing communications. The importance of targeted and personalised messages from relevant medias was a repeating theme as was the customer-centric approach in marketing communication activities. Also, offline and online tools and channels were seen as something that should be treated together as an entity rather than as separate activities. Relevant content created according to the needs of the customer with the gathered data from analytics were seen as the future of b2b marketing communications. Online has added its input to the more traditionally perceived tools and they are executed within the framework of the digital age. Nevertheless, even though online has increased its presence in the b2b marketing communications mix, the more traditionally perceived marketing communication tools, especially personal selling, have not lost their meaning or place in the b2b marketing communication mix.

Fundraising and investor relations in social enterprise: Marketing your way out of the pioneer gap in African off-grid renewable energy

Social enterprises apply the best of business for the pursuit of social or environmental mission while also generating revenues. Globally, nearly 1,3 billion people lack access to electricity, as well as another billion having access to only low quality and infrequent electricity. Off-grid renewable energy, like solar, will increasingly have a key role in the solution of the energy access issue. The pioneer gap in off-grid renewable energy consists of financing (or funding) gaps and capacity gaps, to do with both the early stage of the enterprises in question, as well as the early stage of the whole industry. The gaps are emphasised by specific characteristics of off-grid renewable energy business models and the requirements of operating in bottom-of-the-pyramid markets. The marketing perspective to fundraising is chosen to uncover the possible role enterprises themselves have in bridging the pioneer gap. The purpose of this thesis is to study how social enterprises operating in off-grid renewable energy in Africa utilise marketing activities in their investor relations in bridging the pioneer gap. This main research question is divided into the following sub-questions:  How does the pioneer gap affect fundraising for these enterprises?  How are the funding needs for these enterprises characterised?  How do these enterprises build trust in their investor relations? The theoretic framework is built on relationship marketing and investor relations, with an emphasis on creation of trust. The research is conducted as a thematical case study. Primary data is gathered via semi-structured interviews with six solar energy companies and two accelerators. According to the findings, the main components affecting trust-creation are diminished information asymmetry and perceived risk, mission alignment as well as a personal fit or relationship with the investor. Therefore, an enterprise can utilise e.g. the following marketing activities in their investor relations to bridge the pioneer gap: ensuring investor material, the enterprise story and presenting of them is clear, concise and complete to “package” the enterprise as an investment; taking investor needs and motivations into account as well as utilising existing investors as ambassadors.

Dexamethasone-induced reactivation of bovine herpesvirus type 5 latent infection in experimentally infected rabbits results in a broader distribution of latent viral DNA in the brain

Bovine herpesvirus type 5 (BHV-5) is a major agent of meningoencephalitis in cattle and establishes latent infections mainly in sensory nerve ganglia. The distribution of latent BHV-5 DNA in the brain of rabbits prior to and after virus reactivation was studied using a nested PCR. Fifteen rabbits inoculated intranasally with BHV-5 were euthanized 60 days post-inoculation (group A, N = 8) or submitted to dexamethasone treatment (2.6 mg kg-1 day-1, im, for 5 days) and euthanized 60 days later (group B, N = 7) for tissue examination. Two groups of BHV-1-infected rabbits (C, N = 3 and D, N = 3) submitted to each treatment were used as controls. Viral DNA of group A rabbits was consistently detected in trigeminal ganglia (8/8), frequently in cerebellum (5/8), anterior cerebral cortex and pons-medulla (3/8) and occasionally in dorsolateral (2/8), ventrolateral and posterior cerebral cortices, midbrain and thalamus (1/8). Viral DNA of group B rabbits showed a broader distribution, being detected at higher frequency in ventrolateral (6/7) and posterior cerebral cortices (5/7), pons-medulla (6/7), thalamus (4/7), and midbrain (3/7). In contrast, rabbits inoculated with BHV-1 harbored viral DNA almost completely restricted to trigeminal ganglia and the distribution did not change post-reactivation. These results demonstrate that latency by BHV-5 is established in several areas of the rabbit's brain and that virus reactivation leads to a broader distribution of latent viral DNA. Spread of virus from trigeminal ganglia and other areas of the brain likely contributes to this dissemination and may contribute to the recrudescence of neurological disease frequently observed upon BHV-5 reactivation.

Contemporary marketing in the high-tech industry: Perceptions of Finnish IT marketing professionals

Over the last 30 years, new technologies and globalization have radically changed the way in which marketing is conducted. However, whereas their effects on business in general have been widely discussed, the focus of the effects on marketing remains without clear recognition. Global research has been made to shed light onto the issue, but it has widely concentrated on the views of executives as well as the consumer markets. In addition, a research gap is existent in applying the concept of marketing change in a specific business-to-business (B2B) industry. Therefore, the main research question this study seeks to answer is: “How is contemporary marketing conducted in the high-technology industry?” In this research, the researcher considers the specific industry of high-technology. However, as the industry is comprised of differing markets, the focus will be given to one of the industry’s prime sectors – the information technology (IT) markets, where companies offer other firms products or services manufactured with advanced technology. The growing IT-market is considered of critical importance in the economies of technologically ready countries such as Finland, where this research is also conducted. Through multiple case studies the researcher aims to describe how the changes in technology, customer engagement and future trends have shaped the way in which successful high-tech marketing is conducted in today’s marketplace. Then, results derived from the empirical research are presented to the reader with links to existing literature. As a conclusion, a generalized framework is constructed to depict and ideal marketer-customer relationship, with emphasis on dynamic, two-way communication and its supporting elements of customer analytics, change adaptation, strategic customer communication and organizational support. From a managerial point of view, the research may provide beneficial information as contemporary marketing can yield profitable outcomes if managed correctly. As a new way to grasp competitive advantage, strategic marketing is much more data-driven and customer-focused than ever before. The study can also prove to be relevant for the academic communities, while its results may act as inspiring for new focus on the education trends of future marketers. This study was limited to the internal activities done at the high-tech industry, leaving out the considerations for co-marketing, marketing via business partners or marketing at other B2B-industries.

A search for RNA insertions and NS3 gene duplication in the genome of cytopathic isolates of bovine viral diarrhea virus

Calves born persistently infected with non-cytopathic bovine viral diarrhea virus (ncpBVDV) frequently develop a fatal gastroenteric illness called mucosal disease. Both the original virus (ncpBVDV) and an antigenically identical but cytopathic virus (cpBVDV) can be isolated from animals affected by mucosal disease. Cytopathic BVDVs originate from their ncp counterparts by diverse genetic mechanisms, all leading to the expression of the non-structural polypeptide NS3 as a discrete protein. In contrast, ncpBVDVs express only the large precursor polypeptide, NS2-3, which contains the NS3 sequence within its carboxy-terminal half. We report here the investigation of the mechanism leading to NS3 expression in 41 cpBVDV isolates. An RT-PCR strategy was employed to detect RNA insertions within the NS2-3 gene and/or duplication of the NS3 gene, two common mechanisms of NS3 expression. RT-PCR amplification revealed insertions in the NS2-3 gene of three cp isolates, with the inserts being similar in size to that present in the cpBVDV NADL strain. Sequencing of one such insert revealed a 296-nucleotide sequence with a central core of 270 nucleotides coding for an amino acid sequence highly homologous (98%) to the NADL insert, a sequence corresponding to part of the cellular J-Domain gene. One cpBVDV isolate contained a duplication of the NS3 gene downstream from the original locus. In contrast, no detectable NS2-3 insertions or NS3 gene duplications were observed in the genome of 37 cp isolates. These results demonstrate that processing of NS2-3 without bulk mRNA insertions or NS3 gene duplications seems to be a frequent mechanism leading to NS3 expression and BVDV cytopathology.

Evaluating Performance of Content Marketing: YouTube Marketing in Finnish Food and Drink Industry

The purpose of this research was to define content marketing and to discover how content marketing performance can be measured especially on YouTube. Further, the aim was to find out what companies are doing to measure content marketing and what kind of challenges they face in the process. In addition, preferences concerning the measurement were examined. The empirical part was conducted through multiple-case study and cross-case analysis methods. The qualitative data was collected from four large companies in Finnish food and drink industry through semi-structured phone interviews. As a result of this research, a new definition for content marketing was derived. It is suggested that return on objective, or in this case, brand awareness and engagement are used as the main metrics of content marketing performance on YouTube. The major challenge is the nature of the industry, as companies cannot connect the outcome directly to sales.

Recombinant E2 glycoprotein of bovine viral diarrhea virus induces a solid humoral neutralizing immune response but fails to confer total protection in cattle

Two recombinant baculoviruses were produced in order to obtain a bovine viral diarrhea virus (BVDV) immunogen: AcNPV/E2 expressing E2 glycoprotein, and AcNPV/E0E1E2 expressing the polyprotein region coding for the three structural proteins of BVDV (E0, E1, and E2). Mice were immunized with Sf9 cells infected with the recombinant baculoviruses in a water in oil formulation and the production of neutralizing antibodies was evaluated. Since E2 elicited higher neutralizing antibody titers than E0-E1-E2 polyprotein, it was selected to immunize cattle. Calves received two doses of recombinant E2 vaccine and were challenged with homologous BVDV 37 days later. The recombinant immunogen induced neutralizing titers which showed a mean value of 1.5 ± 0.27 on the day of challenge and reached a top value of 3.36 ± 0.36, 47 days later (84 days post-vaccination). On the other hand, sera from animals which received mock-infected Sf9 cells did not show neutralizing activity until 25 days post-challenge (62 days post-vaccination), suggesting that these antibodies were produced as a consequence of BVDV challenge. Even when no total protection was observed in cattle, in vitro viral neutralization assays revealed that the recombinant immunogen was able to induce neutralizing antibody synthesis against the homologous strain as well as against heterologous strains in a very efficient way.

Differential gene expression profiles of hepatocellular carcinomas associated or not with viral infection

Chronic hepatitis B (HBV) and C (HCV) virus infections are the most important factors associated with hepatocellular carcinoma (HCC), but tumor prognosis remains poor due to the lack of diagnostic biomarkers. In order to identify novel diagnostic markers and therapeutic targets, the gene expression profile associated with viral and non-viral HCC was assessed in 9 tumor samples by oligo-microarrays. The differentially expressed genes were examined using a z-score and KEGG pathway for the search of ontological biological processes. We selected a non-redundant set of 15 genes with the lowest P value for clustering samples into three groups using the non-supervised algorithm k-means. Fisher’s linear discriminant analysis was then applied in an exhaustive search of trios of genes that could be used to build classifiers for class distinction. Different transcriptional levels of genes were identified in HCC of different etiologies and from different HCC samples. When comparing HBV-HCC vs HCV-HCC, HBV-HCC/HCV-HCC vs non-viral (NV)-HCC, HBC-HCC vs NV-HCC, and HCV-HCC vs NV-HCC of the 58 non-redundant differentially expressed genes, only 6 genes (IKBKβ, CREBBP, WNT10B, PRDX6, ITGAV, and IFNAR1) were found to be associated with hepatic carcinogenesis. By combining trios, classifiers could be generated, which correctly classified 100% of the samples. This expression profiling may provide a useful tool for research into the pathophysiology of HCC. A detailed understanding of how these distinct genes are involved in molecular pathways is of fundamental importance to the development of effective HCC chemoprevention and treatment.

Measuring Effectiveness of Marketing Message: The Role of Verbal, Visual and Brand Elements

The aim of this paper is to study the role of verbal, visual and brand elements while meas-uring effectiveness of marketing message. The thesis is written in the context of mobile gaming industry. The object of the study is marketing message. To achieve the aim, the main research question was formulated: How do the elements of marketing message, such as verbal, visual and brand, affect the consumer’s attitude toward the ad, emotional response and attention capture? The theory development chapter lays on three corner stones – analysis of previous litera-ture on marketing message and its elements, namely verbal, visual and brand; overview of literature on attitude formation and particularly attitude toward the ad. In addition, investiga-tion of key points of emotional response and attention capture literature finalizes the chap-ter. The empirical part consists of experiment, conducted with 27 participants. Experiment includes the self-report semantically anchored scale, measuring the attitude toward the ad, as well as autonomic measures – eye tracking (attention capture) and facial expressions (emotional response). The results of the experiment showed that the size of the brand element – the logo – has an effect on the attention capture and the overall attitude toward the ad. The bigger the logo, the more time people spend viewing it, and they realise the message is more educa-tional and factual. The measure related to the visual element – the visual complexity – in-creases the intensity of participant’s facial expression. While the measure of verbal ele-ment – the contrast between text and background colours – leads to a better attitude to-ward the ad. The higher the contrast between text and background, the more known the message appears to the viewer.

Organization of the marketing communication activities after fusion - the integration and the decision making processes

This master’s thesis was made in order to gain answers to the question of how the integration of the marketing communications and the decision making related to it in a geographically dispersed service organization could be improved in a situation where an organization has gone through a merger. The effects of the organizational design dimensions towards the integration of the marketing communications and the decision making related to it was the main focus. A case study as a research strategy offered a perfect frames for an exploratory study and the data collection was conducted by semi-structured interviews and observing. The main finding proved that from the chosen design dimensions, decentralization, coordination and power, could be found specific factors that in a geographically dispersed organization are affecting the integration of the marketing communications negatively. The effects can be seen mostly in the decision making processes, roles and in the division of responsibility, which are affecting the other dimensions and by this, the integration. In a post-merger situation, the coordination dimension and especially the information asymmetry and the information flow seem to have a largest affect towards the integration of the marketing communications. An asymmetric information distribution with the lack of business and marketing education resulted in low self-assurance and at the end in fragmented management and to the inability to set targets and make independent decisions. As conclusions it can be stated, that with the organizational design dimensions can the effects of a merger towards the integration process of the marketing communications to be evaluated.

THE ROLE OF CULTURAL AWARENESS IN BUILDING CHINESE HEALTH FOOD BRANDS Using Netnography in Searching Cultural Arguments to Be Used for Marketing Health Foods in China

Despite the complexity of the Chinese culture consumer product businesses should apply them in building brands for Chinese markets. There are reasons to believe that cultural values and attitudes affect the buying behavior of Chinese consumers. Companies that wish to create brands in China should therefore be aware of the prevailing cultural values and consumer attitudes. This thesis will examine which values and attitudes mostly affect Chinese consumers of health food products. The study will be done by conducting a netnography. Because netnography is actually a collection methods rather than a single method, other auxiliary methods will also be applied. These methods are emotion, language and sentiment analysis (ELS analysis). Emotion analysis will be conducted because cultural values are mostly built on emotional basis. Sentiment analysis will assist in recognizing the key factors that help to locate values and attitudes. Because the netnography will be conducted in Chinese web forums by a non-native researcher, linguistic aspects should also be analyzed in parallel with emotions and sentiment analysis. The study shows that the Chinese consumers of health food products put much importance on functional, analytical and collectivistic attitudes as well as social and psychological values. Of all the twelve cultural values defined, the role of family rose above all. Also perseverance, frugality, guanxi and harmony were highly presented. The attitudes were found by recognizing certain attitude factors. Of all the factors, health foods’ functional benefits and aesthetic content together with consumers’ value consciousness surpassed other factors. Besides these results that can be applied by foreign health food companies willing to enter Chinese consumer markets, also academia can benefit this new approach for conducting ethnographies online.

Impact of increased environmental awareness on product marketing

This Bachelor thesis studies how companies use a variety of different sustainable marketing approaches to sell their green products. Empirical part is done by researching and interviewing two B2B furniture companies.