*4.5. Subcellular Localization*

The coding region of *ZmWRKY106* was fused to the subcellular localization vector p16318h with green fluorescent protein (GFP) tags containing the CaMV35S promoter. The specific primers are listed in Supplementary Table S1. For transient expression assays, the p16318h-*ZmWRKY106* reconstruction plasmid was transformed to maize mesophyll protoplasts by the PEG-mediated method, while the p16318hGFP vector was transformed as control [52]. The fluorescence signals were observed by a confocal laser scanning microscope (LSM700; CarlZeiss, Oberkochen, Germany) after incubation in darkness at 22 ◦C for 18 h.


**German** families were generally characterized by lower level of TV equipment (almost a quarter said that they do not have a TV set) and also by lower television use, which was probably offset by computer equipment that was also used to watch TV programmes.5 The equipment of the other media devices was rather higher than in the other countries. Computers, televisions and surprisingly tablets were more of a parental media, while gaming consoles were used more by children. DVDs, video players and IPods + MP3 players roughly evenly intervened into both generations. In Germany, the daily use of media was also mostly connected with television and computer, about 1/3 of households used social networks every day (Eurobarometer). Germany was a country with a higher level of trust in radio, television and press, but with much lower trust in relation to the Internet and electronic social networks (there was the trust in these tools the lowest from all the analysed countries). PC and Internet gaming was rather smaller than in other countries. The German families were mostly in favour of the statement that the electronic media they have at home allow them to be independent and that the media facilitate communication. When compared to other countries, Germany belonged also among those countries where it was stressed more than anywhere else that the media distance family members away from each other. This rather individualizing aspect that we could observe in German families was confirmed games together.by watching TV together as a family or playing computer.

**Poland** was a country with relatively high media saturation of households; it even showed the highest Internet access (99%). At the same time, the Eurobarometer survey showed a higher level of trust in the Internet and electronic social networks (42% and 28%) in comparison to other countries. The trust was lower in traditional media such as radio, television and press. Television, video and DVD players were strong parental media in the Polish households, while computers, game consoles, IPods, Mp3 players and tablets were clearly used more by children. The daily use of selected media devices was rather lower, in terms of watching TV, videos, DVDs, and playing games on PC and on the Internet. Therefore, it is possible that overall the time devoted to media activities in Poland was lower than in other countries. In terms of attitudes towards the media, the Polish families have identified themselves most with the statement claiming that the media facilitate communication and allow

5For example, they used highly the Internet connection to watch the news (Eurobarometer).

them to pursue their hobbies and entertainment. Watching TV together as a family was quite common, gaming also appeared as an activity that family members do at least occasionally together.

**Slovak families** were well equipped with media devices at the time of the survey, with the exception of game consoles and tablets. In the Eurobarometer survey, Slovak respondents showed rather higher levels of trust in selected media and they watched television mainly through a TV set (not via the Internet). According to Eurobarometer, Slovakia was one of the countries with lower daily computer use (52%), but this was not reflected in the sample of our families. Slovakia reported more than 90% of daily computer and television use. However, substantially lower was a daily use of other media devices. In this respect, Slovak households have more traditional focus. As a daily leisure time activity, watching TV and computer use were represented in about 60% of respondents. In comparison to other countries, in Slovakia was recorded higher computer and Internet gaming, around 50% of households play games every day. However, it was rather an independent activity of respondents, not a joint playing of family members. On the other hand, watching television was more common activity of households. Slovak respondents of our research strongly inclined to the opinion that the electronic media they have facilitated their communication and negotiation. This was related to the fact that thanks to the media they could be more independent. Slovak families where those of all studied countries who also agreed the most to spend more time with the media than talking at home with other family members (53%).

**Ukrainian** households6 were also relatively well equipped with the media, only gaming consoles in more than 3/4 of households were not present. In the Ukraine, television and computers were used extensively every day, as well as IPod, MP3 players, gaming consoles and mainly tablets. Tablets were also used in the most of the Ukrainian households, with almost three quarters of those who use these tablets every day. IPods, MP3 players, computers and tablets were more of a children's media, because more than 50% of them were used by children. In the Ukrainian households, there was also a higher frequency of computer games, 80% said that they were playing daily and almost 50% that it was even more than 2 hours a day. For the Ukrainian respondents, the media mainly meant that everyone could devote their time to hobbies and entertainment and that they could spend their free time together. However, almost 50% of respondents also reflected that they usually spend more time with the media than talking at home, and almost one quarter also reported frequent quarrels at home because of the media.

The families we studied had in common a considerable saturation of some media devices—computers, the Internet access, mobile phones and also TV sets. On the other hand, some of the devices have not been so common in the households (tablets, game consoles). Some of the recent digital devices showed differences at the time of our survey; for example, IPods and MP3 players were already present in most of the households in the Czech Republic, Germany, Slovakia and also in Ukraine, but in

6The Ukraine was not part of the Eurobarometer 88 survey; therefore, the data come only from our LSCF survey.

Latvia and Poland the prevailing part of households did not have them. Video players were widely represented in the Czech Republic and Slovakia, while in the other countries it did not occurred in most of the households. Televisions and computers were used mostly every day; this was same for all the countries. Differences were found mainly in the case of the daily use of tablets, IPods and MP3 players. Tablets were mostly used mainly in the Ukraine and Latvia, IPods and MP3 players every day in more than 50% of households, that owned them, in the Ukraine.

Overall, it can be stated that the media and media facilities are an essential part of the lives of contemporary families and that they have been well represented in households at the time of our survey. At the same time, the development in this area has progressed significantly in recent years and it can thus be assumed that the current modern digital means are also largely involved in the leisure time activities of family members. Some media activities are carried out in families together, some rather encourage individualization within family life. This is one of the reasons why the interviewed families point out the benefits that the media bring to their lives, that the media allow them to be more independent and that they also allow easy communication between family members and bring a way how to realize their hobbies and entertainment. Nevertheless, the media in some households are also a source of tension, conflict and distance family members from each other. However, these negative aspects did not the interviewed families highlighted as strongly as the apparent positives of the media.

#### **References**

Watkins, S. C. (2009). *The young & the digital*. Boston: Beacon Press.

**Open Access** This chapter is licensed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license and indicate if changes were made.

The images or other third party material in this chapter are included in the chapter's Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the chapter's Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.

### **Conclusion**

Family represents topic that is extremely serious and up to date. Is has undergone significant changes as never before over the last decades. Despite all these changes, which has undergone, it remains primary educational institution and primary social environment. All this complexity we have tried to show in this publication, how difficult it is to deal with traditional functions, how standard of living is influenced by the contemporary society, how actually the contemporary family looks like and lives. Both on the basis of all available expert knowledge from not only sources, but also from confrontations with foreign ones, based on several surveys and mainly above all our research.

In the introductory chapter, we present two basic categories of the whole monograph: the family and the way of living. There is a description of the post-war family development in Europe until now, differentiated in the Western and Eastern part. It turns out that changes in the society have caused significant changes also in the family lives. Furthermore, we explain the term lifestyle and its different concepts, ways of lifestyle and its types according to various aspects. At present, the concept of healthy lifestyle and its shaping plays a highly significant role.

In the review study, there are several surveys which deal with family lives (e.g. Maˇríková 1999, Chaloupková 2005, Kraus and Jedliˇcková 2007, Viš ˇnovský et al. 2010, Holubová 2011). They relate to roles in the family and their changes, division of work, household work, public view at marriage and parenthood, parents' needs, issues in the family and time budget. In the conclusion of the whole chapter, there is project of our survey, its goals, methods, research sample (total of 2437 respondents) and the survey process. The research team set a goal to monitor family lifestyle in four fields: socioeconomic situation, life satisfaction, leisure time and ways of spending it and media in a family, its use and attitudes towards its role in the family lives.

The contemporary family is more likely in all countries somehow marked by advancing democratization, social differentiation, disintegration and by unemployment to varying degrees, certain isolation and intergenerational relationships passes through certain changes as well.

Authors also pay attention to family social policy, housing situation when starting a family, the fact, that in individual countries is done by the society. Also, an increase

B. Kraus et al., *Contemporary Family Lifestyles in Central and Western Europe*, SpringerBriefs in Sociology, https://doi.org/10.1007/978-3-030-48299-2

in violence as well as differences in family lives living in towns or in the country side is stated. The image of a German family, unlike other countries, is marked by strong migration wave.

The social economic situation seems to be a big current issue. Tomu je vˇenována následující kapitola. This is documented in the international research, monitoring life conditions in households in the Czech Republic, Poland, Hungary and Slovakia and are confirmed by the results of our international research. We are interested in the income situation, expenditure items, experience with unemployment and respondents were also to evaluate their overall standard of living. The Germans and then the Czech respondents evaluate their situation as the best. The worst perception is given by families from Latvia.

We also monitored the overall life satisfaction. According to the respondents, it is represented by harmony living without conflicts, well-being, good health of all family members and material security without major differences shown among monitored countries. On the other hand, the differences were found in cases where respondents had to state what they lack to satisfaction. Most often, it was financial security and the lack of free time for the family.

Historical context and changes are described in the chapter about leisure time in terms of amount and ways of spending leisure time. The importance of leisure time has gradually increased for individuals and also for the society. The current leisure time is no longer perceived quantitatively (such as time, that is left), but based on its quality, such as time of freedom, recreation and self-realization. It fulfils several important functions for the human. In most surveyed countries, the lack of mutually spent leisure time with the family was perceived as the most important issue that hinders family satisfaction. In terms of the amount of leisure time, the most satisfied respondents come from Latvia and the Czech Republic. In these countries, the leisure time outweighs the standard working week, weekends and holidays. On the contrary, the least satisfied with the amount of free time are respondents from Poland. In terms of quality of spending of leisure time, the function of relax is essential for working respondents. It is evaluated in a positive way. The number of respondents who are satisfied with the quality of relax in their free time outweighs significantly. But this is no longer true for the satisfaction of the time of their interests. The respondents would like to make more trips, travel, do more sport, social games and fun with their families. They are most often prevented from two reasons—lack of time and finance. Reading and watching television are dominant leisure time activities of individual respondents. Whereas reading is clearly individual activity, watching television is an activity that is shared by family members. Other leisure activities in which the whole family participates are gardening, visiting friends and relatives, visits of cultural events, walks and board games.

At the time of the research, the households in the surveyed countries were already saturated with electronic media. Families with children are generally the types of households that obtain new technologies quickly and use them. They often get new media tools because of children and to use them for school preparation in order to "keep up with" their peer groups. The families were very well equipped with computers, mobile phones, televisions and also had the Internet access. On the other hand, some tools were being found in households (e.g. game console or IPod and MP3 players mainly in Latvia and the Ukraine). What, was possible to observe at the same time in our data, was a certain "generational differentiation" of media, and who uses them at home most often. For example, while the television was dominant parent medium, then tables, game consoles, IPod and MP3 players were used by children to a much greater extend.

Media activities were among relatively frequent activities of leisure time members of the survey families. Especially, watching television and computer activities belonged to what the families do every day. Watching TV together was then favourite activity of a large part of Slovakian, Polish and Latvian households. Family members point out both benefits and negatives as well what media brings to them. All in all, they point out more positives brought by the media. They perceive media device as a way of easier communication, independence in order to pursue hobbies and fun.

In our research, there were included five countries of the former Eastern block of countries and one from the Western part. In many aspects of family lives, it was possible to distinguish differences between both groups. On contrary, there were found also dissimilarities among individual countries. We hope that our data can mirror very interesting features of lifestyles in studied countries and show the lived diversity in Europe.

*The investigation is carried out within the project "Development and support of a multidisciplinary scientific research team for studying the current family at UHK" - CZ. 1.07/2.3.00/20.0209. This publication is supported by the Faculty of Education, University of Hradec Králové.*

## **Appendix**

#### **Questionnaire**

*We would like to ask you to complete an anonymous questionnaire focused on the quality of life of contemporary families. Its completion is not time consuming—should not take more than 20 minutes. Please tick with a cross x chosen possibility. For items where are more possibilities given tick all of them which you prefer. For items where an answer is necessary to be completed—just write down your opinion very briefly.*

#### **1. Permanently live in :**

#### **5. The highest level of education**:

#### Appendix 117

#### **15. Express yourself to the satisfaction with the quantity and quality of the free time.**

#### **16. What activities would you like to do in your free time?**

……………………………………………………………………………………………………………………… ……………………………………………………………………………………………………………………

**17. Do you play sometimes together PC games at home?**

YES, very often YES, occasionally NO, never

**18. Do you do any sport in your free time?** YES NO

#### Appendix 119

#### **19. If you do sport, what sport do you do? How often and which of them do you do together with a**

**family member?** *(write down and please tick the box with the suitable answer*:

#### **20. Define the scope of your daily free time in hours:**

**21. Each of us assumes some areas in the life that are more important than others and tries to seek for achievement. Which areas you consider to prefer in your family – put into order according to your importance from the most important "1" to less important "9"** *fill in number into the boxes)***.**

Healthy, safe environment

	- Others *(write): ………………………………………………………………………*

#### **22**. **What leisure activities are typical for you?** You can list up to 5 answers*. In case of chosen*

*activity please list who you do it with.*

#### **23. What types of electronic media do you have in your household and what number?**

*(list the number of each media)*

#### **24. Who from your family uses this equipment the most?** *(please tick the box with x)*

#### **26. To what extent to you agree with these following statements?** *(please tick the box with x)*

#### **27. Do you sometimes watch TV together?**

YES, very often YES, occasionally NO, never

#### **28. If yes, what type of TV program do you usually watch together?** *(write down)*

*………………………………………………………………………………………………………………………… …………………………………………………………………………………………………………………………*

#### **29. What is the main reason that you watch TV together?**

We have no more other televisions, there is no other way. We like the same programme so we watch it together. We like to watch together; therefore we choose the same program. From other reason *(write*): ………………………………………………………………..

#### **30. How often in your free time do you spend these following activities?**

Appendix 125

**31.** If you have something to say, to add to the investigation *(write down)*

*………………………………………………………………………………………………………………………… ………………………………………………………………………………………………………………………… …………………………………………………………………………………………………………………………*

*For the team of investigators—thank you for your time and willingness while completing the questionnaire!*


**3.2 L-ascorbic acid**

*Lactose and Lactose Derivatives*

**3.1 Acetic acid**

US\$11.39 billion by 2022 [3, 73–75].

sustainable growth of 5.48% AAGR (average annual growth rate) in the last years and it is expected that it could increase globally up to US\$9.29 billion by 2021 and

Biosynthesis of an OA is obtained by the biochemical pathway of cellular metabolism, as the final end product or as an intermediate product of a path [26]. Bacteria and fungi are the most available and suitable living organisms for the industrial production of OAs. The microbial production of organic acids is usually an attractive route for industrial implementation compared to chemical synthesis because the conditions used in microbial bioprocesses tend to be less extreme (in terms of temperature, pressure, extreme pH) and more friendly to the environment [3, 76]. However, this may be effective only if the concentration of these acids in the fermentation broth are high enough (in the order of tens or hundreds of grams per liter), and these are obtained in reasonably short times [77]. Also, the microbial bioconversion of sugars into organic acids is frequently carried out by strict anaerobic microorganisms, with relatively long fermentation, reduced productivity, and low titers of organic acids in the fermentation broth [27]. Those facts conspire with its large-scale implementation, and to turn the biotechnology in an economically

attractive choice to the production of organic acids (**Figure 2**) [3, 26, 78].

scarce are the microorganisms that can use lactose to achieve this.

Acetic acid (C2H4O2, MW 60.052 g mol−1

(18%) are the largest producers [84, 85].

In this context, the processes of isolation and purification of organic acids become critical [78, 79]. Various alternatives for the isolation and purification of organic acids from fermentation broth or biomass have been used. Among the most used primary purification methods are precipitation with Ca-salt or hydroxide [77], ammonium salt, organic solvents [80], and ionic solutions [81]. Microbial fermentation can produce directly only a few organic acids [74], and even more

monocarboxylic acid commonly used as a chemical starting reagent in the production of important chemicals, like cellulose acetate, polyvinyl acetate, and synthetic fibers. Vinegar (near 4% vol. acetic acid) is produced by fermentation of different carbon sources by acetic acid bacteria [82] and is widely employed in food preparation and cooking since ancient times. Currently, three-quarters of the world production is obtained by carbonylation of methanol (by chemical synthesis), basically from nonrenewable sources, while 10% is still obtained from the microbial biotransformation of sugars [83]. By 2014, the global acetic acid market reached 12,100 ktpa, with an average price of US\$ 550 per ton and average annual growth of 4–5% [14]. In 2018, world production reached 16,300 ktpa, near to US\$ 12.48 billion, forecasting production of 20,300 ktpa by 2024. China with 54% and the US

*Worldwide production of some organic acids between 2018 and 2025. (A) High-, (B) medium -, and*

, IUPAC name: Ethanoic acid) is a

**56**

**Figure 2.**

*(C) low-level of global production.*

A case is the L-ascorbic acid (vitamin C, C6H8O6, MW 176.124 g mol−1 , IUPAC Name: (5R)-[(1S)-1,2-Dihydroxyethyl]-3,4-dihydroxyfuran-2(5H)-one), one of the organic acids with the highest production and sales volumes today. Vitamin C can be obtained by microbial biosynthesis but from D-sorbitol [86].

Ascorbic acid, previously called hexuronic acid, is a soluble white solid and organic compound that presents itself as two enantiomers: L-ascorbic acid (vitamin C), and D-ascorbic acid, without any biological role found [87, 88]. Vitamin C is an essential nutrient for humans and many animals, and its deficiency can cause scurvy, in the past a common disease among sailors in long sea voyages [89]. It is used in as a food additive and a dietary supplement for its antioxidant properties [87, 88]. There is a report, however, that achieves the synthesis of vitamin C from the lactose present in the cheese whey, but through a defined group of chemical reactions [90]. In 2015 was produced 150.2 ktpa of ascorbic acid with a revenue of US\$820.4 million. By 2017, China produced near to 95% of the world supply of vitamin C, having revenue of US\$880 million [91].