2.2.3. COVID-19 Worries

Participants were asked about their worries regarding the pandemic (e.g., contracting COVID-19, transmitting COVID-19 to someone else, family/friends contracting COVID-19, having enough food, and loss of income). Five response options were provided ranging from not at all, a little, a moderate amount, a lot, to a great deal. Responses were collapsed into three categories, not at all (not at all), somewhat (a little, a moderate amount), and a lot (a lot, a great deal) for analyses.


**Acknowledgements**

plants could act as antinutritional factors, thereby reducing the bioavailability of important nutrients and the nutritional value of the food. According to Kokhar and Apenten [51], this effect is present a result of a selected adaptive mechanism due to a "chemical warfare" between

**Figure 3.** KEGG Pathway database diagrams presenting: (A) Inositol Metabolism; (B) Galactose Metabolism, showing the Raffinose Family Oligosaccharides (highlighted in red). Green Box: enzymes with transcriptional modulation data available from stress assays. Red Box: enzymes without transcriptional modulation data available in the literature. Yel‐

Among the various Ins-derivatives, phytic acid (1-D-myo-inositol-P6; Figure 1) is the most studied, concerning the impacts on human and animal health. Its unique structure provides the ability to chelate cations such as iron, zinc, potassium, magnesium, and copper, forming

higher plants and herbivorous pests.

92 Abiotic and Biotic Stress in Plants - Recent Advances and Future Perspectives

low Box: enzymes not covered in this review.

The authors thank CNPq (National Council of Technological and Scientific Development – process number 454112/2014-9) for financial support. JRCFN thanks to CAPES (InterSis Network – Biocomp Project) for the fellowship granted. EAK thanks to CNPq for the fellowship granted.


**3. Results and discussion**

having a diameter of about 2.5 cm, and thickness of about 0.4 cm is attained. Then, the annealed

36 Applications of Laser Ablation - Thin Film Deposition, Nanomaterial Synthesis and Surface Modification

**Figure 1.** Photograph of different experimental methods demonstrating (a) homogeneously pelletized ceramic target, (b) high-temperature annealed target, (c) cleaning of substrates, (d) target loading, (e) loading of substrates, (f) impurities removal in UHV chamber (glow discharge), (g) laser ablation, (h) target after ablation, (i) thin films after deposi-

The procedure for predeposition cleaning of substrates and growth of nano-thin phosphor

The morphology of the product was analysed by field emission scanning electron microscope (FESEM-SUPRA 55). Using atomic force microscopy (NTEGRA PRIMA-NTMDT, USA), the surface topography of the thin phosphor films was studied. The crystal structure and phase purity of as-synthesized phosphor were recognized and confirmed by PANalytical's X'Pert PRO Materials Research X-ray Diffractometer (Almelo, USA) equipped with a CuKα radiation (λ = 0.154060 Å) at a scanning rate of 0.02°s−1 in a 2*θ* range of 15°–60°. Further, down-conversion PL excitation and emission studies and fluorescence decay time measurements were performed at room temperature using a Cary bench-top spectrophotometer (AGILENT Instru-

films were already discussed in detail on our previous work [2].

target is used for laser ablation.

tion.

**2.2. Characterization**

ments, USA).


**5. Conclusion**

In Eastern Siberia lyssavirus isolated for the first time from a bat (*Murina leucogaster*) was reported in September, 2002. The bat, which looked healthy, entered into the house in Ir‐ kutsk. It was caught and observed. On the tenth day of its observation the symptoms of dis‐ ease appeared which has resulted in fatal outcome.  During the strain isolation the infected suckling mice had 18 days of the incubation period, and among these mice only one fell ill. At the second passage the incubation period varied from 9 to 18 days. Moreover, it should be noted that the bat infected with Irkut strain wounded the girl, but the illness did not de‐

Both cases in Irkutsk and Primorye Territory confirm that usually sick animals enter into hu‐ man dwellings. First of all such animals are of a special danger and require to take measures which protect people from the possible accidental contacts with them. Other cases of lyssa‐ virus infection in Europe reported by Botvinkin et al. (2003) look very similar to these ones. Unfortunately, the final diagnosis in studied case of infection was stated postmortem. The isolation of Ozernoe strain revealed the following facts to pay attention to: a short incuba‐ tion period up to 6-7 days; the examined suckling mice of a mouse family fell ill at the same time. If the suckling mice were challenged intracerebrally, the virus titre in the brain of the dead patient was evaluated as 3.7 lg LD50. In the first passage the titre of Ozernoe strain in the mice challenged intracerebrally reached 6 lg LD50.  These data indicate a high virus concentration which caused profound brain damages and the overall visceral in‐ jury followed by impairment of vital functions and resulted in the imminent fatal outcome. The postmortem analysis indicated the changes characteristic for rabies: extensive destruc‐ tive changes of neurons, formation of many neuronophagic nodules like "rabies nodules", an evident spongy endema of the medullary substance. At the same time, a clearly defined in‐ flammatory reaction around blood vessels was not found. This fact together with the found changes in lymphoid organs indicated the immunodeficiency state, which was undoubtedly

The presence of extracellular viral particles with morphology similar to the structures of Rabies virus on cell surface membranes and spreads forming the electron-dense sub‐ stance in cellular cytoplasm were revealed during the ultrastructural study. Extracellular viral particles in the environment were distinctly observed and had the morphological signs typical for the group of enveloped viruses with spikes on the surfaces [Iseni, 1998]. The oval and dense bodies (with diameter about 2550 nm) were determinated in cyto‐ plasm and mainly in extracellular environment of PEG cells culture infected by strain Ozernoe. These structures were described earlier as Negri bodies in neurons infected by

To identify exactly the virus genotype the fragment of N gene was amplificated and se‐ quenced. Moreover, the search for homologous sequences in GeneBank using BLAST pro‐ gram has given the absolute evidence that the closest relative of Ozernoe strain is Irkut strain isolated earlier from the bat in Eastern Siberia. Since the Ozernoe strain is the first strain of the genotype 8 isolated from a dead human, we have determined its complete ge‐

velop due to the timely vaccination against rabies.

244 Encephalitis

associated with the virus infection.

Rabies virus [Velandia et al., 2007].

The clinical and epidemiological characteristics of fatal human case of lyssavirus infection identified fort he first time in the Asian part of Russia were provided. The evidence that this case belongs to Lyssavirus infection in the terms of etiology was obtained based on the data of epidemiological anamnesis, clinical picture of infection as well as the virological, morpho‐ logical, and molecular genetic studies. The pathologic diagnosis was the acute stage of me‐ ningoencephalitis as underlying disease with complications of edema, swelling and dislocation of brain, the formation of neuronophagic nodules like "rabies nodules" mainly in the subcortical brain as well as bilateral hypostatic pneumonia and parenchymatous degen‐ eration of myocardium, liver and kidney. The study by electron microscopy revealed that the viral particles of 100 nm in diameter could also be associated with the vesicles close to the endoplasmic reticulum or Golgi apparatus. The complete genome sequence of Ozernoe strain is 92% identical to the complete genome of Irkut strain and 77-78% to EBLV-1 ge‐ nome. The phylogenetic analysis based on the complete genome sequences revealed that Ozernoe strain, isolated from the brain of the dead patient, and Irkut strain, isolated in Ir‐ kutsk from the brain of a bat, are located on the same branch of the phylogenetic tree and have the common ancestor. So the real possibility of serious Lyssavirus infection in cases when people have accidentally encountered with chiropteran was demonstrated.

This case has ended in fatal outcome because of the fact that doctors have never registered patients with a lyssavirus infection in the Far East before. Therefore, it was difficult to make the right decision for emergency vaccination against rabies after the accidence as it is usually done in the cases of classical rabies. The exact diagnosis was not get, the clinical course of infection was rapid and following remedial actions have been unsuccessful.

[5] Botvinkin, A. D., Selnicova, O. P., Antonova, L. A., et al. (2005). Human Rabies case

The Fatal Case of Lyssavirus Encephalitis in the Russian Far East

http://dx.doi.org/10.5772/52869

247

[6] Boulger, L. R., & Porterfield, J. S. (1958). Isolation of a virus from Nigerian fruit bats.

[7] Bourhy, H., Kissi, B., Lafon, M., Sacramento, D., & Tordo, N. (1992). Antigenic and molecular characterization of bat rabies virus in Europe. J Clin Microbiol., , 30,

[9] Familusi, J. B., Osunkoya, B. O., Moore, D. L., Kemp, G. E., & Fabiyi, A. (1972). A fa‐ tal human infection with Mokola virus. Am. J. Trop. Med. Hyg., , 21, 959-63.

[10] Fekadu, M., Shaddock, J. H., Sanderlin, D. W., & Smith, J. S. (1988). Efficacy of rabies vaccines against Duvenhage virus isolated from European house bats (Eptesicus se‐

[11] Felsenstein, J. (1985). Phylogenies and the comparative method". American Natural‐

[12] Fraser, G. C., Hooper, P. T., Lunt, R. A., Gould, A. R., Gleeson, L. J., Hyatt, A. D., Russell, G. M., & Kattenbelt, J. A. (1996). Encephalitis caused by a lyssavirus in fruit

[13] Fooks, A. R., Mc Elhinney, L. M., Pounder, D. J., et al. (2003). Case report isolation of a European bad lyssavirus type-2a from a fatal Human case of Rabies Encephalitis. J.

[14] Gribencha S.V.(2008). Rabies. In: Lvov D.K., editor. Meditsinskaya Virusologia (Med‐

[15] Hanlon, C. A., De Mattos, C. A., De Mattos, C. C., Niezgoda, M., Hooper, D. C., et al. (2001). Experimental utility of rabies virus-neutralizing human monoclonal antibod‐

[16] Heaton, P., Johnstone, P., Mc Elhinney, L., Cowley, R., O'Sullivan, E., & Whitby, J. (1997). Heminested PCR assay for detection of six genotypes of rabies and rabies re‐

[17] Hillis, D. M., & Bull, J. J. (1993). An empirical test of bootstrapping as a method for assessing confidence in phylogenetic analysis. Systematic Biology , 42, 182-192.

[18] Hughes, G. J. A., reassessment, of., the, emergence., time, of., European, bat., & lyssa‐

[19] Iseni, F., Barge, A., Baudin, F., Blondel, D., & Ruigrok, R. W. H. (1998). Characteriza‐ tion of rabies virus nucleocapsids and recombinant nucleocapsid-like structures. J.

ical Virology) [in Russian]. Moscow: "Med. Inform. Agenstvo":, 586-594.

ies in post-exposure prophylaxis. Vaccine , 19, 3834-3842.

lated viruses. J. Clin. Microbiol., , 35, 2762-6.

virus, type. . (2008). Infect. Genet. Evol., , 8, 820-824.

rotinus), classic rabies and rabies-related viruses. Vaccine , 6, 533-539.

caused from a bat bite in Ukraine. Rab.Bull.Europe., р. 3.

Trans. R. Soc. Trop. Med. Hyg., , 52, 421-4.

bats in Australia. Emerg. Infect. Dis., , 2, 327-31.

Med. Virol.;71, , 2(2), 281-289.

Gen. Virol., , 79, 2909-2919.

[8] Rab, Bull., & Europe, . (2008). (1-4), 1-4.

2419-26.

ist , 125, 1-15.


*4.2.2. Alternatives to ITO*

**Figure 7.** The plot of resistivity improvement versus time for various TCOs.

by various solution-based deposition processes is given in **Table 3**.

**Table 3.** Properties of ITO films prepared by various solution-deposition processes.

 **(Ω)** *t* **(nm)** *T* **(%) ΦH (10−2 Ω−1) Deposition Ref**

45.7 47.5 144 85.3 0.420 Electrospray [79] 42.3 188 225 80 — Dip coating [76] 2.5 42 60 ∼85 0.469 Dip coating [81] 2.1 7.1 295 ∼78 1.171 Dip coating [82] 310 — 75 >90 — Spin coating [80] 0.52 356 146 93 0.136 Spin coating [74] 4.2 81 70 — — Spin coating [77] 7.2 30 241 90.2 1.191 Spin coating [72]

ITO is the most widely used TCOs because of its two key characteristics: conductivity and optical transparency. Conventionally, ITO films are produced by vacuum-based deposition techniques. The best resistivity of ITO film deposited by sputtering was reported as low as 1.2 × 10−4 Ω cm [70]. As an alternative to vacuum-based deposition, recently much efforts have been focused on production of solution-based deposition for high-performance ITO films using various approaches such as nanomaterials-based [71–75], aqueous metal salts solution [76], combustion synthesis [77, 78], and advanced annealing techniques (microwave annealing [79] and ultraviolet laser annealing [80]. A summary of properties of ITO films prepared

**ρ (10−4 Ω cm)** *R***s**

**4.2. N-type conductive oxides**

*4.2.1. Indium-tin-oxide (ITO)*

88 Green Electronics

There are increasing emphases on not only developing a TCO with higher conductivity but also with a material other than ITO. Two major concerns with ITO are the cost and the potentially limited resource.

Current candidate materials for a higher conductivity TCO are summarized in **Table 4**, which include the conventional binary oxides of CdO, SnO2 , In2 O3 , and ZnO, with alternative dopants, and combinations of these binaries, plus Ga2O3, in ternary, quaternary, and even quinary combination of these five oxides.

As a promising binary TCO material, ZnO is considered the prime candidate for a higher conductivity and to be low cost. However, ZnO is more sensitive to oxygen than ITO, and process control is more difficult. Work continues on improving ZnO with alternative dopants, especially Al (AZO) and Ga (GZO). As for sol-gel AZO film, the lowest reported resistivity is 1.5 × 10−4 Ω cm [83], which is close to that of film prepared by sputtering [84].

**Figure 8.** (a) Cross-sectional structure and (b) transfer curves of ZIZO-TFT using SCS-ITO as S/D electrodes.

below 300°C. We then used lanthanide elements (Ln, except Ce) to stabilize the amorphous phase. As a result, we found a series of solution-processable amorphous p-type conductive Ln-M-O (a-Ln-M-O, where M = Ru, Ir, and Ln is lanthanide elements except Ce) having low resistivity (10−3–10−2 Ω cm) (**Figure 9**). The resistivity increases with increasing Ln atomic number (Z) and changes evidently faster for Z > 64 (Gd), which may be more related to the influence of 4f electrons (4f subshell is more than half-filled at Z > 64) than to the decreasing size of Ln ions. These oxides are thermally stable to a high degree, being amorphous up to 800°C, and processable below 400°C. These oxides have three pronounced features: first, their

2*g*

Ru6

tively different from the closed-shell or pseudo-closed-shell configurations in other p-type

Second, the conduction of amorphous La-Ru-O (semiconducting) is completely different from

resistivity of solution-processed a-La-Ru-O is lower than that of the sputtered sample. Hence, detailed understanding of the electronic structure of these materials and the mechanism underlying processing-composition-conduction correlation is of fundamental importance for

The unique properties of amorphous p-type oxides have high potential for use in printed electronics, e.g., as gate electrode in n-type oxide TFTs [33, 94], and may be modified to use as p-channel in TFTs or as an active layer in solar cells. The narrow band-gap is advantageous for light absorbing in solar cells. The electronic configurations of these amorphous oxides are apparently not analogous to those of known p-type oxides. This deserves theoretical investi-

of A element. These compounds are semiconducting for A = Y and the lanthanides Pr-Lu, and weakly metallic for A = Bi and Pb. The Ln of the aforementioned oxides was thus replaced with Bi and Pb, leading to low-resistivity amorphous oxides produced at low

O11 and La4

in SnO, Rh *t*

4 in Ru 4d and *t*

Low-Temperature Solution-Processable Functional Oxide Materials for Printed Electronics

2*g*

2*g*

6 in Zn-Rh-O, and Co *t*

Ru2 O7

5 in Ir 5d), which is distinc-

2*g*

, is sensitive to the type

O19, which are both metallic). Third, the

http://dx.doi.org/10.5772/intechopen.75610

6 in Zn-Co-O).

91

valence electrons have open-shell configurations (*t*

in Cu oxides, Sn 5s2

Ru3

gation and suggests that more p-type amorphous oxides are ahead of us.

It is known that the conduction of ruthenium pyrochlore, A2

**Figure 9.** (a) XRD patterns and (b) resistivity of LnRuO films.

oxides (e.g., Cu 3d10so

future research.

that of crystalline phases (e.g., La3

**Table 4.** Conductive oxides based on multinary metal oxide systems other than ITO.


**2. Results**

#### *2.1. Chemical Composition of Essential Oils*

Thyme and oregano essential oils are, generally, characterized by a large amount of monoterpenes (both hydrocarbons and oxygenated), reaching almost 90% of the whole oil. The two main phenolic monoterpenes, thymol and carvacrol, occur more frequently, often accompanied by their biogenetically correlated compounds *p*-cymene and γ-terpinene [13,24]. Thymus is probably one of the most taxonomically complex genera of its family. Several studies confirmed that the two most common chemotypes are thymol and carvacrol [25],

followed by less common non-phenolic chemotypes [26]. As reported in materials and methods section, the two EO samples of this study were commercial. The chemical composition of our oregano sample has already been published in a previous study with different purposes from this one [27], while the chemical composition of our commercial thyme EO is published here for the first time. The two analytical results are listed together in Table 1 for easier comparison by readers. The samples subjected to this study had chemical profiles quite typical of the *Origanum vulgare* thymol and carvacrol chemotype and the *Thymus capitatus* carvacrol chemotype of essential oils. Gas chromatography (GC) techniques, coupled with a flame ionization detector (FID) and a mass spectrometer (MS), allowed the identification of more than 35 components, covering up to 99% and 98% of the total oil compositions, respectively. Table 1 shows the details of the chemical compositions, listing only the components with a % > 0.05. The most represented class for both samples was that of oxygenated monoterpenes (68.7% and 72.9%, respectively), followed by hydrocarbon monoterpenes (30.0% and 22.3%, respectively). The sum of these classes in the oregano sample reached 96.7%, while in the thyme sample it reached 95.1%. For both samples, sesquiterpenes and other components were below 4%. The common feature of these two oils was to have three main components which alone covered more than 80% of the whole composition. The oregano essential oil profile was characterized by the presence of carvacrol (36%) and thymol (25%) as the main compounds, followed by *p*-cymene (22%). All other compounds were below 5%. The thyme essential oil profile was dominated by a high amount of carvacrol (almost 70%), followed by *p*-cymene (9%) and γ-terpinene (8%). The thymol percentage was negligible (0.5%), as were the percentages of all other compounds, with the exception of β-caryophyllene, which reached 2.6%.

**Table 1.** Chemical composition of commercial *Origanum vulgare* and *Thymus capitatus* essential oils (EOs).

**Table 1.** *Cont.*

a The numbering refers to the elution order. b Literature retention index (RI). c Retention index (RI) relative to the standard mixture of *n*-alkanes on the SPB-5 column. d Identified compounds (those < 0.05% have not been reported). e Data previously reported (see [27]). f Relative peak area percent, representing the averages of three determinations. N.D. = not detected.

#### *2.2. Physicochemical Characterization of Essential Oil-Loaded Nanocapsules (EO-NCs)*

In a previous work, we reported the encapsulation of thyme and oregano essential oils in PCL nanocapsules [28]. Unlike the previous work, here we used commercial essential oils having different compositions of volatile components. This offered numerus advantages, including the lower cost of preparation and the standardization in the compositions of the EOs. In this manner, the obtained nanocapsules could potentially be more attractive for industrial sectors. Table 2 shows the reported values of the physicochemical parameters (z-average diameter, polydispersity index (PDI), zeta potential (*ζ*), encapsulation efficiency (EE) and loading capacity (LC)) which allowed us to characterize the nanocapsules containing thyme and oregano essential oils (TC-NCs and OV-NCs, respectively).

**Table 2.** Physicochemical Characterization of essential oil (EO)-NCs.

The z-average diameters of 198 ± 3 nm and 200 ± 3 nm for the TC-NCs and OV-NCs, respectively, were in agreement with the nanometric structures of the nanocapsules. Low PDI values for both nanocapsules pointed out a narrow size distribution of nanoparticles and the presence in aqueous solution of monodisperse nanosystems (Figure 1). Negative zeta potentials of −11 and −10 mV for the TC-NCs and OV-NCs, respectively, were very similar to those obtained for other stable PCL nanocapsules [28,29]. The percentages of EE and LC were high, with values of 84 ± 6 and 52 ± 3 for the TC-NCs and 80 ± 9 and 51 ± 4 for the OV-NCs. The prepared TC-NC and OV-NC aqueous nanosuspensions showed a total essential oil content of 5.7 ± 0.3 and 5.8 ± 0.4 mg/mL, respectively.

**Figure 1.** I-weighted distribution of the hydrodynamic diameter (*DH*) of (**a**) OV-NCs and (**b**) TC-NCs.

#### *2.3. Antifungal Activities of Pure EOs and EO-NCs*

The screening of encapsulated essential oils (EO-NCs) against a panel of fungal strains (Table 3) showed marked antifungal activity (Table 4 and Figure 2). The obtained results evidenced that EO-NCs inhibited the growth of assayed fungi at s MIC values ranging from 0.125 to 0.25 mg/mL. These concentrations were two to four times lower than those observed for pure EOs. For both EO-NCs, an MIC of 0.125 mg/mL was effective for inhibiting the growth of *A. fumigatus*, *C. aggregatocicatricatum*, *C. herbarum* and *P. eryngii*. In addition, this concentration for OV-NCs was also enough to inhibit the fungus *Bjerkandera adusta*.

#### **Table 3.** Characteristics of the assayed fungal strains.

IMB-SAS = fungal collection of the Institute of Molecular Biology (Slovak Academy of Sciences). CCBAS = Culture collection of Basidiomycetes, Institute of Microbiology, Academy of Sciences of the Czech Republic.

We also evaluated the fungicidal effect of EO-NCs versus the panel of fourteen fungal strains. In particular, both the TC-NCs and OV-NCs had minimum fungicidal concentrations (MFCs) of 0.25 mg/mL against *A. fumigatus*, *C. aggregatocicatricatum*, *C. herbarum* and *P. eryngii*, while having an MFC of 0.5 mg/mL against *A. flavus*, *P. rubens*, *P. citrinum*, *F. oxysporum*, *G. candidum*, *M. circinelloides*, *E. xenobiotica*, *P. lilacinum* and *P. chrysosporium*. The MFC values of the EOs were two to four times higher than the MFCs of the EO-NCs on the tested strains. The empty NCs revealed no activity on the fungal growth.

**Table 4.** Minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values of encapsulated essential oils and pure essential oils.

MIC and MFC are expressed in mg/mL. TC-NCs = nanoencapsulated thyme essential oil; OV-NCs = nanoencapsulated oregano essential oil; TC-EO = thyme essential oil; and OV-EO = oregano essential oil.

**Figure 2.** Example of MIC and MFC assays with the isolate *Cladosporium herbarum*. (**A**) Results using a nanoencapsulated oregano EO. (**B**) Results using a nanoencapsulated thyme EO.C=control. The range of nanoencapsulated EO concentrations used was 0.05–0.5 mg/mL.


**1. Introduction**

Human evolutionary studies or paleoanthropological research are constantly yielding new information and thus revising previously assumed hypotheses as well as generating new ones. While Africa and Europe have dominated the bulk of our knowledge on human evolution over the last century, various parts of Asia are yielding new and unexpected paleoanthropological surprises. One of these vital Asian regions is South Asia or the Indian Subcontinent, its prehistory being known and regularly highlighted since the nineteenth century [1] and predominantly includes stone tool assemblages from various time periods ranging from the Lower Paleolithic to the Neolithic [2]. Prehistoric evidence is known from throughout the Subcontinent with specific geographic pockets as being exceptions due to various

factors including research bias as well as other natural attributes. Lithic assemblages belonging to all prehistoric phases have been reported including Lower Paleolithic (Oldowan and Acheulean), Middle Paleolithic, Upper Paleolithic and microlithic/ Mesolithic. Despite this large body of known evidence, very few sites have been properly dated using absolute dating techniques. The earlier results, though obtained through different dating methods [3, 4], should be viewed as provisional until verified by newly-available dating techniques. For example, some U-Th dates (between <390 Ka and < 131 Ka) processed a few decades ago at a multi-period site in Rajasthan have now been revised to younger estimates using the luminescence method e.g. [5], leading to a re-interpretation of that cultural sequence [6]. The persistent marginal profile of hominin fossils continues to afflict Indian prehistory and more systematic surveys are required to identify new areas with vertebrate fossil preservation. The only known pre-modern hominin fossils in the subcontinent, which may be contemporary with the Late or terminal Acheulean phase, come from Hathnora and nearby localities in the central Narmada Valley. They include a partial calvarium, possibly female, and possibly associated clavicles and a rib fragment, all

#### **Figure 1.**

*Map of dated Paleolithic technologies across the Indian subcontinent including Pakistan, India, Nepal and Sri Lanka (LP: Lower Paleolithic; a: Acheulean; EA: Early Acheulean; LA: Late Acheulean; MP: Middle Paleolithic; UP: Upper Paleolithic; M: Microlithic; MC: Multi-cultural; OES: Ostrich eggshell).*

**37**

**Table 1.**

*List of dated prehistoric sites in the Indian subcontinent.*

*Human Evolution in the Center of the Old World: An Updated Review of the South Asian…*

**Site Age Techno-chronology** Masol 2.6 Ma? pre-Acheulean Riwat (Pakistan) ~2 Ma Pre-Acheulean Pabbi Hills (Pakistan) 2.2–0.9 Ma Pre-Acheulean

Attirampakkam 1.5 Ma & 385–73 Ka Acheulean & Middle Paleolithic

Isampur 1.27 Ma? Acheulean Singi Talav ~800 Ka? Acheulean Dhansi >780 Ka Undiagnostic Morgaon >780 Ka & 41 Ka Acheulean Dina & Jalapur ~700–400 Ka Acheulean Chirki Nevasa >350 Ka Acheulean Sadab 290 Ka Acheulean Teggihalli 287 Ka Acheulean Umrethi >190 Ka Acheulean Kaldevanhalli 174 Ka Acheulean Patpara & Bamburi 1 140–120 Ka Acheulean Bori 1.38 Ma to 23 Ka Acheulean Adi Chadi Wao 190 to 69 Ka Acheulean 16R (Didwana) 187 Ka - 6 Ka Multi-period Sandhav 114 Ka Middle Paleolithic Bhimbetka Rockshelter III-F23 >106 Ka & >41 Ka Multi-period Nakjhar Khurd >100 Ka Acheulean Durkadi <100 Ka Multi-period Kataoti 95 Ka Middle Paleolithic Dhaba 79–65 Ka & 48 Ka Middle Paleolithic & microlithic Jwalapuram 77–38 Ka & 35 Ka Middle Paleolithic and microlithic

recovered over a decade [7, 8]. The calvarium, originally identified as an "advanced" *Homo erectus*, was later reclassified as an archaic or early form of *H. sapiens* [9, 10]. Phylogenetic reevaluation of the calvarium reveals that it shares key morphological

Mehtakheri 48 Ka Microlithic Fa-Hien Lena (Sri Lanka) 48 Ka microlithic Kalpi 45 Ka Middle Paleolithic Site 55 (Pakistan) 45 Ka Upper Paleolithic Kitulgala beli-Lena (Sri Lanka) 45 Ka microlithic Sanghao Cave (Pakistan) 42 Ka Middle Paleolithic Kana 42 Ka Microlithic Batadomba Lena (Sri Lanka) 36 Ka microlithic Mahadebbera 34 Ka Microlithic Arjun 3 (Nepal) >30 Ka? Middle Paleolithic Patne 30 Ka Multi-period

*DOI: http://dx.doi.org/10.5772/intechopen.94265*

*Human Evolution in the Center of the Old World: An Updated Review of the South Asian… DOI: http://dx.doi.org/10.5772/intechopen.94265*

### **Table 1.**

*List of dated prehistoric sites in the Indian subcontinent.*

recovered over a decade [7, 8]. The calvarium, originally identified as an "advanced" *Homo erectus*, was later reclassified as an archaic or early form of *H. sapiens* [9, 10]. Phylogenetic reevaluation of the calvarium reveals that it shares key morphological

features with both *H. heidelbergensis and H. erectus* [11]; it has been most recently classified as *Homo* sp. indet. [12]. The oldest fossil evidence for *Homo sapiens* is dated to ~38 Ka and currently comes from Sri Lanka, while all younger evidence comes from multiple sites across India [13, 14].

What is also largely missing is direct evidence for butchery in the form of cutmarked fossil bones; some possible exceptions include Isampur [15] and Masol [16], both of which require further verification and substantiation through more evidence. Additionally, use-wear analyses and other scientific methods such as residue analysis are also required on well-preserved lithic assemblages. Other types of evidence that are poorly known is the age and nature of symbolic behavior (see [17]) as well as the nature of technological transitions. Indeed, there has been a recent global movement to decolonize earlier interpretations of hominin dispersals and population replacements across the Old World [18]. This also includes India, where earlier historical interpretations defined the Upper Paleolithic and modern human behavior based on the then-known European evidence [19]. Numerous reviews of the South Asian region's prehistoric records have been published elsewhere (e.g. [3, 4, 20–28]). Over three dozen Paleolithic and early microlithic sites have been dated in Pakistan [29–33], India [5, 14, 16, 34–58], Nepal [59] and Sri Lanka [60–63] since the 1980s onwards, using different relative and absolute dating methods including biochronology, palaeomagnetism, stratigraphic correlation, U-Th, U-series, K-Ar, Ar-Ar, luminescence, electron spin resonance, radiocarbon (calibrated and uncalibrated) and AMS. These various ages range from ~2.6 Ma to ~35 Ka, and include geographically random sites belonging to various prehistoric technologies including Oldowan-like, Acheulean, Middle Paleolithic, Upper Paleolithic and the earliest microlithic assemblages (**Figure 1** and **Table 1**). While broad summaries are provided here, the primary goal of this paper is to highlight the most salient attributes of this zone, provide specific updates to previously known data and discuss possible implications of new discoveries from surrounding regions outside the Subcontinent.


Preface

The catastrophic earthquake sequence in Türkiye and Syria in February 2023 is a reminder that many aspects of earthquake science need further multidisciplinary research. Scientists and engineers must understand the fault's rupture and estimate the earthquake's ground motion and its consequences on building performance. In other words, a new educational curriculum must merge the seismology, geotechnical, and structural engineering fields to fulfill research trends and demand practices. This book attempts to start to do so.

*Earthquakes - Recent Advances, New Perspectives and Applications* results from the tireless effort of thirteen distinguished scholars from eight countries and is organized into four sections containing nine chapters.

Section 1, "Tectonics and Seismicity", includes Chapters 1–4. In Chapter 1, Motazedian and Ma study the June 23, 2014, M 7.9 Rat Islands Alaska earthquake by applying a full moment tensor inversion and a source rupture model accompanied by hypocenter relocations of aftershocks. The chapter can be used to teach how to perform inversions at universities and research centers for engineers and seismologists. In Chapter 2, Bogdanov and Pavlov construct probabilistic models that make it possible to trace self-consistent changes in space, time, and energy for background earthquakes preceding a strong event (e.g., M > 7.0) by employing wavelet decomposition methods, including depth uncertainty, in the analysis. The authors use the earthquake catalog for the Kamchatka regional network of the Russian Academy of Sciences during the period 1994–1997. In Chapter 3, Salazar presents a methodology to test the Weibull and Poisson probability distribution of earthquakes, employing a clustered and declustered catalog based on subduction events in El Salvador; magnitude conditional probabilities are computed for a time-dependent seismic hazard assessment. In Chapter 4, Motazedian and Ma recalculate the original parameters of the intraplate 1982 Miramichi earthquake and aftershocks using a master-event location method combined with regional depth-phase modeling.

Section 2, "Earthquake Forecasting", includes Chapter 5 in which N. Solarić and M. Solarić present a prominent application of GPS satellites for earthquake forecasting, analyzing the distance changes amongst different Earth stations before and after an earthquake occurrence.

Section 3, "Geotechnical Engineering", includes Chapter 6, in which Hsu presents an improvement of the classical methodologies to calculate the bearing capacity on soils considering the strain softening and asymmetric shear failures under the finite element analysis. It also presents new findings on Coulomb's earth pressures and slope stability analysis.

Section 4, "Building Structures", includes Chapters 7–9. Chapter 7 by Apak summarizes the morphology of the vernacular structures in Türkiye at Adana City and presents a qualitative analysis of these heritage buildings due to vertical, seismic, and wind loadings, including out-of-plane behavior. Chapter 8 by Alomari describes the risks associated with modifying the structural systems of buildings. It also shows remedial measures locating special devices to reduce the earthquake response of the structures. Chapter 9 by Trcala, Němec, and Gálová explains several structural dynamics techniques employing accelerograms as input excitation of buildings in the nonlinear range behavior.

> **Walter Salazar, Ph.D.** Professor, Faculty of Engineering and Architecture, Catholic University of El Salvador UNICAES, Santa Ana, El Salvador

> > Section 1

Tectoncis and Seismicity

Section 1


*Edited by Ifeoluwapo O. Amao*

*and Iyabo B. Adeoye*

This book discusses pertinent aspects of agricultural economics and rural development. It includes case studies that assess the impact of tomato cultivation on food security and poverty alleviation of rural dwellers and agricultural producers. Additionally, it examines farmers' knowledge of organic livestock farming, a novel method of livestock production. The book also contains a review of factors affecting the efficiency of vegetable production and the basics of good agribusiness plans for successful agribusiness activities.

ISBN 978-1-83968-537-8

Agricultural Economics

Published in London, UK © 2021 IntechOpen © Aleksej Filatov / iStock


**2.1 Classification of ECs**

ECs are classified into organic, inorganic micro-pollutants like pesticides, personal care products (PCPs), pharmaceuticals, synthetic organic dyes, polycyclic aromatic hydrocarbons (PAHs), heavy metals ions, plasticizers, per-fluorinated compounds, flame retardants, surfactants, etc. (**Figure 1**) generated by human activities such as domestic, health care units, agricultural and industrial pathways [10]. These compounds are a source of concern due to their physical and chemical properties because they are widely distributed in the environment which is harmful to humans and wildlife. These pollutants are difficult to detect and have varied activities and miscellaneous sources of production. Their presence in small concentrations causes chronic toxicity, endocrine disruption, and the expansion of pathogen resistance [11].

**Figure 1.** *Classification of emerging contaminants.*

### *2.1.1 Pesticides*

Pesticides, a class of organic contaminants, based on their physical and chemical properties are categorized as fungicides, herbicides, bactericides, and insecticides which are used in the agricultural sector to control dangerous insects, weeds, and microorganisms, etc. Based on their application sites, pesticides are frequently detected in groundwater causing toxicity and may bio-accumulate in humans and plants, or sediments depending on solubility, reactivity, and characteristics of soil and environment. Among the pesticide contaminants, dichlorodiphenyltrichloroethane (DDT) and hexachlorocyclohexane are commonly used pesticides (about 67%) as compared to other compounds such as phorate, chlorpyriphos, Atrazine, methyl parathione, Bentazone, Diazinon, Cyanazine, Simazine, phosphamidone, Terbuthylazine, Alachlor and Dimethoate [12].

### *2.1.2 Pharmaceutical industry*

Pharmaceuticals are major emerging organic contaminants occurring in small amounts in water resources worldwide [13]. Pharmaceuticals are extensively used on daily basis in human healthcare as well as veterinary medicine such as nutrition, investigative aids, therapy and preventive medicine. Many pharmaceutical products such as drugs (both prescribed and non-prescribed), hormones and antibiotics are extensively detected in the aquatic environment, surface and groundwater and have adverse effects on humans, poultry, livestock and fish farming, etc. Generally, livestock is given medications to reduce diseases and infections. Researchers have examined more than 3000 chemicals used in therapeutic products but only small proportion (ng L−1 doses) has been studied in the field, which possibly will lead to negative effects on human and wildlife. To enhance animal farming, organic fertilizer such as manure and purines as medicines are used which indirectly affect the atmosphere and can reach living organisms through food stuff. Commonly reported pharmaceuticals in wastewater are antibiotics, diclofenac, antacids, clofibric acid, steroids, antidepressants, ciprofloxacin, propranolol, beta blockers, analgesics, salicylic acid, fluoxetine, antipyretics, anti-inflammatory drugs, nitroglycerin, tranquilizers, lipid-lowering drugs and stimulants [14].

Natural or synthetic hormones are also essential ecological contaminants, because of their estrogenic and androgenic impacts on wildlife. Organic and inorganic hormones consist of 17α-estradiol, 17β-estradiol, estrone, equiline, equilenin, estriol, mestranol and norethindrone which can enter atmosphere through farming, and are not completely eliminated from wastewater and harm aquatic life and humans.

### *2.1.3 Personal care products (PCPs)*

Personal care products (PCPs) are household chemicals commonly used for health, odor, beauty, or cleaning. These chemicals are used in personal care products like ornamental cosmetics, soaps, hair and skin care products, lotions, fragrances and sunscreens. PCPs are used in large quantities throughout the world due to which the release of these pollutants in the environment is increasing day by day [15]. Mostly these substances are bioactive and bioaccumulative and harm the environment and humans [16]. The most probable emerging contaminants in PCPs are antiseptics, perfumes pollutants like galaxolide, pest repellants, preservatives diethyl phthalate ultraviolet (UV) filters and Triclosan (TCS) and triclocarban as disinfectant

### *Technologies for Removal of Emerging Contaminants from Wastewater DOI: http://dx.doi.org/10.5772/intechopen.104466*

pollutant. Parabens are antimicrobial preservatives used in cosmetic items, pharmaceuticals, and some food stuffs such as benzyl, butyl, ethyl, isobutyl, isopropyl, methyl, and propyl hydroxybenzoates. Polycyclic musks are used in numerous products such as clean-up products, shampoos, hair care and washing products and cosmetic products. Their use on the outside of human skin increases its discharge in environment without any metabolic changes. Among all these products, cosmetics are frequently used, thus its occurrence in air at low quantity may be a source of damage to human beings, wildlife and environment.

### *2.1.4 Surfactants*

Surfactants are synthetic organic compounds used all over the world in making of household products such as emulsifiers, detergents, paints, and pesticides, in addition to personal care products and are harmful to aquatic species [17]. They are classified as cationic, anionic and zwitterionic surfactants. Frequently used surfactants such as fatty alcohol ethoxylates, linear alkyl benzene sulfonates, lignin sulfonates, and alkyl phenol ethoxylates are produced on a large scale. Furthermore, octylphenol and nonyl-phenol ethoxylates, are highly toxic even at low concentrations and must be substituted in all their uses.

### *2.1.5 Food additives*

Numerous artificial sweeteners such as acesulfame, saccharin and sucralose which are extensively used in foodstuff, pharmaceuticals and hygiene products find their way to domestic wastewater via human excretion. These moderately metabolized sweeteners which pollute the environment are usually hard to remove. Though, latest calculated ratio for predicted environmental concentration (PEC) and predicted no effect concentration (PNEC) of compound sucralose for marine system is below 1 indicating limited threat to aquatic system (plants, algae and fish) [18].


**1. Introduction**

148 Advances in Unconventional Lithography

In summary, we designed and fabricated two types of binary diffractive convex lenses using EBL on a PET film. In the case of the binary diffractive convex lens with 2-mm focal length, it is possible to control the luminous intensity distribution. To improve the diffraction efficiency and realize a thin LED light source, we designed a binary diffractive lens with 140-μm focal length. This type of lens with focal wavelengths in the micrometer range can

To realize the binary diffractive lens with the 100-μm-order focal lengths, we characterize the relationship between the diffractive lens structure and its light intensity. It is clear that the intensities of the zero- and first-order diffractions are controlled by the structure of the binary diffractive lens. By using this lens, wide luminous intensity distribution can be obtained.

This work was partly supported by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (No. 18360008 and 21360007), Mie University COE Projects, a Start-up Grant Program for Mie Venture Business from Mie Industry Enterprise Support Center, the Kinki Invention Center, and the Knowledge Cluster Initiative from the

The authors thank Mr. T. Sato (JA Woollam Japan Co., Inc.) for ellipsometry measurement. The authors also thank Dr. H. Miyake, Mr. K. Manabe, Mr. N. Machida, Mr. Y. Nakayama, Mr. K. Arakawa, and Mr. Y. Seriguchi for their help in the experiments and valuable discussions.

Jordan, J. A. ; Hirsch, Jr. P. M. ; Lesem, L. B. & Van Rooy D. L. (1970). Kinoform Lenses.

Swason, G. J. & Veldkamp, W. B. (1989). Diffractive optical elements for use in infrared

Orihara, Y. ; Klaus, W. ; Fujino, M. & Kodate, K. (2001). Optimization and Application of

Yamada, K. ; Watanabe, W. ; Li, Y. ; Itoh, K. & Nishii, J. (2004). Multilevel Phase-type

Lalanne, P. ; Astilean, S. ; Chavel, P. ; Cambril E. ; Cambril, E. & Launois, H. (1999). Design

Lalanne, P. (1999). Waveguiding in Blazed-Binary Diffractive Elements. *Journal of Optical*

Mait, J. M. ; Prather, D. W. & Mirotznik, M. S. (1999). Design of Binary Subwavelength

Kleemann, B. H. ; Seesselberg, M. & Ruoff, J. (2008). Design Concepts for Broadband High-

Hybrid-Level Binary Zone Plates. *Applied Optics*, 40, 32, (5877-5885), ISSN0003-6935

Diffractive Lenses in Silica Glass Induced by Filamentation of Femtosecond Laser

and Fabrication of Blazed Binary Diffractive Elements with Sampling Periods Smaller than the Structural Cutoff. *Journal of Optical Society of America. A*, 16, 5,

Diffractive Lenses by Use of Zeroth-order effective-medium Theory. *Journal of*

efficiency DOEs. *Journal of the European Optical Society*, 3, (08015-1–08015-16) ISSN

produce a thin LED light source to control the luminous intensity distribution.

Ministry of Education, Culture, Sports, Science and Technology.

*Applied Optics*, 9, 8, (1883-1887), ISSN0003-6935

(1143-1156) ISSN0740-3232

1990-2573

systems. *Optical Engineering*, 28, 6, (605-608) ISSN0091-3286

Pulses. *Optics Letters*, 29, 16, (1846-1848), ISSN0146-9592

*Society of America. A*, 16, 10, (2517-2520) ISSN0740-3232

*Optical Society of America. A*, 16, 5, (1157-1167) ISSN0740-3232

**4. Conclusion**

**5. Acknowledgment**

**6. References**

Stimuli-responsive polymeric materials are able to change their chemistry and their conformation upon an external signal. The external signal may be derived from a change in temperature, chemical composition or applied mechanical force of the specific material, or can be triggered externally with exposure to an electric or magnetic field or to light irradiation. In this respect, a photochromic substance is a stimuli responsive material which is characterized by its ability to alternate between two different chemical forms having different absorption spectra, in response to light irradiation of appropriate wavelengths (Brown 1971). Due to this important property, a significant amount of effort has been devoted to the formation of polymeric materials functionalized with photochromic molecules for the creation of photosensitive "smart material" systems, that change reversibly their physical and chemical properties by the use of light. The corresponding reversible effects of the molecules such as dipole moment, surface energy, refractive index, and volume are preserved in the polymer matrix, and have numerous promising applications in devices for three-dimensional (3D) optical memories, (S. Kawata & Y. Kawata 2000), in actuators (Yu et al 2003, Athanassiou et al 2005), in holographic or diffractive optics, (Fu et al 2005, Tong et al 2005) or in microfluidics, (Caprioli et al 2007, Walsh et al 2010) etc. Concerning microfluidic devices using photochromic plastic films, the transportation of fluids happens without the need for their molecules to be charged, as done in other studies (Mitchel 2001). This is achieved by gradually modifying the surface tension, and thus the wettability, by irradiating with increasing time along the direction of the fluid movement (Ichimura et al 2000). The gradual wettability changes are exclusively based on the photochemical modification of the embedded photochromic molecules caused by the photoisomerization process. In addition, in the case of the diffraction gratings the development was generally done by interference of different polarized laser beams, or by electric-field application, and the modification of their diffraction efficiency is connected with the changes of the refractive index of the photochromic molecules during this procedure (Yamamoto et al 2001, Fu et al 2005).

Here we present how the volume changes induced to the photochromic polymers by the photoisomerization of their embedded photochromic molecules, can improve significantly

Photocontrolled Reversible Dimensional Changes of Microstructured Photochromic Polymers 151

Briefly, UV and visible irradiation causes the reversible transformation of these chemical species, between two states (isomers) that have light absorption bands in characteristic spectral regions. This property is retained when the photochromic molecules are incorporated in polymer matrices, where they are homogeneously dispersed forming miscible systems. Specifically, the absorption properties of the photochromic polymer films prepared as described above change reversely upon UV-visible irradiation as shown at Figure 2. Initially the system is transparent at the visible range of the spectrum. Upon pulsed UV laser irradiation the SP is slowly converting to the MC isomer, fact indicated by the new absorption band in the visible region of the spectrum (ca. 565 nm). The intensity of the peak increases with the number of UV pulses until a plateau is reached, which suggests that the photoisomerization is completed and that the system has reached the equilibrium. The subsequent irradiation with green laser light, causes the decrease of the intensity of the previously formed MC peak, while after a certain number of pulses the spectrum reaches its initial form, indicating that MC reverts fully to the SP isomer. These data confirm that under the irradiation conditions mentioned in the figure caption of Figure 2, the reversible properties of SP are retained in the host polymer matrix. Depending on the irradiation conditions and the weight percentage of the photochromic molecules in the polymer matrix (usually ≤10%), it has been shown that typically about 4-10 irradiation cycles can be performed, while further irradiation causes the degradative photooxidation of the photochromic molecules, restricting thus the lifetime of the system (Athanassiou et al 2006c). Additionally, the degradative phenomena start to be evident usually after the third cycle. In order to exclude this parameter from the following study, results derived by the first three

Fig. 2. Absorption spectra of the PEMMA/SP 10% wt upon UV and visible irradiation. For the specific study, the irradiation conditions used are: λUV=355 nm, fluence FUV=20 mJ cm-2,

irradiation cycles are presented.

λvis=532 nm, Fvis=35 mJ cm-2.

the performance of these two different type of applications, namely the microfluidic devices and the diffraction gratings. In both cases the lithographic technique used for the microstructuring of the photochromic doped plastic films is the soft molding. Concerning the microfluidics applications, the presented microstructured photochromic plastic films exhibit a significant improvement on the reversible wetting characteristics compared to those on the flat surfaces. This improvement is due to the combination of the changes in the surface polarity and thus in the wetting properties with the modified surface conformation, both provoked by the light induced changes of the photochromic molecules. Moreover, regarding the optical gratings, we present a different approach in where the control of their diffraction efficiency relies on the dimensional variations of the gratings upon laser irradiation. Following this approach, the efficiency of the gratings is significantly improved with respect to previous works.

Such findings open the way for the production of optically switchable gratings based on reversible dimensional changes, and can be of great importance in all-optical signal processing systems. Moreover, the ability to control the wettability of surfaces by microstructuring and to tune it by using photochromic molecules, permits the application of these lithographically formed structures to all-optically controlled switches capable of operating with tunable speed, and to microfluidic actuation.


**1. Master in evolving**

Antibiotics are extensively used worldwide for treating predominantly gram-negative bacterial infections and also for treating certain gram-positive infections. While the precise mechanism of their bactericidal action is yet to be unraveled, aminoglycosides, for example, act by binding to the RNA component of ribosomes, leading to both mistranslation and ultimate inhibition of protein synthesis. The widespread use of other major classes of antibiotics has resulted in the emergence of resistant bacteria by expediting the course of its evolution [1, 2]. The emergence of resistance to antibiotics is of special concern in the treatment of infections, particularly of systemic nature, by gram-negative organisms narrowing down the options for antibiotic alternatives. The resistance mechanisms displayed by the bacteria can be classified into the following: (a) reduced uptake, (b) increased efflux, (c) enzymatic modification of drug, and (d) drug target modification. Whereas resistance to streptomycin, the first widely used aminoglycoside, is predominantly through mutations in drug targets (mostly in the ribosomal protein rpsL and also in rRNA), resistance to other aminoglycosides appears to utilize a variety of mechanisms. The question arises, whether antibiotic action facilitates the emergence of resistant mutants. For certain other classes of antibiotics that induce the bacterial SOS response either by direct DNA damage (e.g., ciprofloxacin) or through indirect means (e.g., ampicillin), it has been shown that the action of the antibiotic itself plays a significant role in the emergence of mutations that confer resistance. One such mechanism, mistranslation due to defects in the translation apparatus, can promote hypermutagenesis in a phenomenon called translational stressinduced mutagenesis (TSM) raising the possibility that aminoglycoside exposure, by promoting mistranslation, could also elevate mutagenesis. According to the current understanding, TSM is mediated by a low-level mistranslational corruption of the replicative DNA polymerase leading to episodic hypermutagenesis. Exposure of wildtype bacterial cells to sublethal concentrations of an antibiotic increases mutagenic translesion DNA synthesis in vivo, and exposure of certain mutants also increases spontaneous mutagenesis. Exposure of wild-type *Pseudomonas aeruginosa* PAO1 cells to sublethal concentrations of tobramycin and amikacin, two aminoglycoside antibiotics commonly used to treat *P. aeruginosa* infections, can elevate spontaneous mutagenesis leading to complications in treating cystic fibrosis patients [3].

### **2. Master in dominating**

Cystic fibrosis (CF) is an autosomal recessive genetic condition among Caucasians, with an incidence rate of 1 in 2500 live births. The morbidity and mortality associated with this disease condition are due to thickened lung secretions and subsequent creation of hypoxia and secondary infections predominantly by opportunistic pathogens. Bacteria such as *Pseudomonas aeruginosa*, *Staphylococcus aureus*, and *Burkholderia cepacia* complex have been in the limelight as the pathogens that affect CF patients with progression of lung disease ultimately leading to mortality. Interestingly, recent developments in high-throughput genomic techniques revealed the presence of several other bacterial species, which were hardly identified using conventional microbiological techniques. Enteric bacteria, such as *Prevotella*, *Bacteroides*, *Fusobacterium*, *Mycoplasma*, *Ralstonia*, *Veillonella*, etc., which do not normally appear in the laboratory cultures were identified and were found to have an impact on the CF microbiome [4]. This fluctuation in the CF microbiome may be due to transition of atypical species toward chronic mode of infection through formation of biofilms, dormancy, small colony variants, etc. The immunocompromised nature of CF patients predisposes them to a variety of infections, thereby increasing the need for antibiotics, alone or in combination, on a daily basis, at milligram levels. Such a continuous antibiotic pressure drives evolution of lung pathogens through the downregulation of acute-mode virulence factors in order to avoid unnecessary energy loss and expression of regulons associated with chronic mode of infection/colonization. Though the CF microbiome has been shown to consist of several species of bacteria, *P. aeruginosa* becomes the predominating one during the course of chronic colonization in the CF lung, thereby increasing its significance when considering appropriate treatment strategy [5].

Apart from the abovementioned bacterial species in the CF microbiome, mycobacteria, in general, are widespread organisms except tuberculosis (*Mycobacterium tuberculosis*) and leprosy (*M. leprae*) pathogens which are obligate parasites always in need of a host. These bacteria are often involved in asymptomatic infections, are highly fastidious organisms showing resistance to antibiotics, and are able to survive for long periods in acids, alkali, detergents, etc. Non-tuberculous mycobacteria constitute all the other mycobacteria gaining importance in respiratory infections including the one resembling tuberculosis. Practically, overgrowth of pseudomonads and other predominant bacterial species in the lung makes it difficult to understand the existence of atypical bacteria in the case of CF lung infections. The inherent slow growth rate of mycobacteria adds to failures in preliminary detection of these bacteria. Once identified it requires a prolonged treatment regime for several months with combined antibiotics, which add stress to the CF lung environment, thereby resulting in a progressive deterioration of lung function with consequent emergence of resistant pathogens. Dominance by *P. aeruginosa* or few known predominant bacterial species in the CF lung is clinically beneficial in the sense that these outnumbered species may offer protection against more pathogenic species such as mycobacteria [6].


**6. Integrated model for androgen action**

Androgens diffuse across plasma membrane due to their lipophilic nature. Once inside the cell, they are recognized by the cytosolic androgen receptor, which forms a complex with its ligand. This complex works as a transcription factor, which translocates to cell nucleus, binds to DNA and promotes the activation of several genes (Beato, 1989). This is called the

Cardiovascular Effects of Androgens 73

depicted in Figure 2, testosterone activates a plasma membrane G-protein coupled receptor, a signaling pathway that results in a nongenomic mechanism. Activation of this pathway facilitates the hydrolysis of phosphatidylinositol 4,5-bisphosphate by phospholipase C, thereby generating diacylglycerol and IP3. IP3 mediates opening of IP3 receptors in the sarcoplasmatic reticulum, which in turn produce rapid (1-5 min) Ca2+ release (Vicencio et al., 2006). Similar effects of testosterone have been identified in T-cells, skeletal muscle and neuroblastoma cells (Benten et al., 1999; Estrada et al., 2003, 2006). In cardiomyocytes, nongenomic signals activate the extracellular signal-regulated kinase (ERK) and the mTOR

pathways. Activation of Ca2+/ERK/mTOR axis is independent of androgen receptor.

regulates gene expression.

**7. Perspective**

protocols.

**8. Acknowledgment**

*Circ Res,* 84, 813-819.

**9. References**

This work was supported by FONDECYT grant 1090276.

The anabolic properties of testosterone indicate that this hormone regulate multiple genes involved in growth and metabolism. Traditionally, ligand-dependent androgen receptors act on DNA within the nucleus and in this compartment there is cross-talk among the nuclear receptors and cytosolic signaling. We have determined that testosterone induce a hypertrophic pattern in cardiomyocytes, evaluated as increase in the expression of fetal proteins, cardiomyocyte size, and protein synthesis. Hypertrophy effects of testosterone were blocked by either inhibition of mTOR or androgen receptor (Altamirano et al., 2009). We propose that cell growth produced by androgens require both androgen receptor activity and translation control through mTOR signaling pathway. Thus, both mTOR pathway and androgen receptor could control protein synthesis by a coordinated mechanism, where mTOR regulates translation and the intracellular androgen receptor

The role of sex steroid hormones in modulating cardiovascular function is of the highest importance, given that androgen deficiency is strongly associated with common medical conditions including metabolic syndrome, obesity, diabetes, hypertension and atherosclerosis. However, cardiovascular side effects of high doses of anabolic/androgen steroid reduce its actual therapeutic use. Research in this field is necessary to know the real cardiovascular effects of androgens and to elaborate future optimal therapeutic replacement

Akkad, A. A.; Halligan, A. W.; Abrams, K.; & al-Azzawi, F. (1997). Differing responses in

Altamirano, F.; Oyarce, C.; Silva, P.; Toyos, M.; Wilson, C.; Lavandero, S.; Uhlen, P.; &

transdermal estrogen replacement therapy. *Obstet Gynecol,* 89, 97-103. Alexandersen, P.; Haarbo, J.; Byrjalsen, I.; Lawaetz, H.; & Christiansen, C. (1999). Natural

blood pressure over 24 hours in normotensive women receiving oral or

androgens inhibit male atherosclerosis: a study in castrated, cholesterol-fed rabbits.

Estrada, M. (2009). Testosterone induces cardiomyocyte hypertrophy through mammalian target of rapamycin complex 1 pathway. *J Endocrinol* 202, 299-307.

classic or genomic androgen response pathway. On the other hand, it has been reported that androgens, exert a rapid, non-genomic effect mediated by a putative plasma membrane receptor. Its activation implicates participation of kinases, intracellular calcium oscillations and transcription factor activation, among other fast events.

Cardiomyocytes are terminally differentiated cells. Cardiac hypertrophy is characterized by an increase in cell size and protein synthesis and reactivation of a fetal gene program. Emerging evidence indicate that androgen action involves a crosstalk between genomic and nongenomic mechanisms. These could imply a two-step process, where both non-genomic and genomic effects occur sequentially (**Figure 2**). First, fast effects would be generated at the plasma membrane level with second messenger participation. Second, there would be gene activation; the pathways used by each cell type could provide specificity to the signal through a cooperative mechanism, which can act in a concerted form.

Fig. 2. Integrated mechanism for testosterone effects in cardiomyocytes. In cardiomyocytes, testosterone activates plasma membrane androgen receptors coupled to heterotrimeric Gq protein. Gβγ dimers activate phospholipase C (PLC), which locally produces inositol IP3 thus activating IP3 receptors (IP3R). The consequent Ca2+ increase leads to the activation of several pathways, including the MEK/ERK/mTOR axis. These responses combined induce a series of genomic changes that lead to increased gene expression and increased protein synthesis, ultimately leading to cardiac hypertrophy.

While studying these hypertrophy effects on cardiomyocytes, our laboratory determined that testosterone uses a cooperative mechanism between the genomic and nongenomic pathways to produce the hypertrophic growth of cardiac cells. Thus, with normal levels of androgens the interactions among signaling pathways lead to the maintained health of cardiac cells, whereas high doses recruit additional signals which are hypertrophic. As

classic or genomic androgen response pathway. On the other hand, it has been reported that androgens, exert a rapid, non-genomic effect mediated by a putative plasma membrane receptor. Its activation implicates participation of kinases, intracellular calcium oscillations

Cardiomyocytes are terminally differentiated cells. Cardiac hypertrophy is characterized by an increase in cell size and protein synthesis and reactivation of a fetal gene program. Emerging evidence indicate that androgen action involves a crosstalk between genomic and nongenomic mechanisms. These could imply a two-step process, where both non-genomic and genomic effects occur sequentially (**Figure 2**). First, fast effects would be generated at the plasma membrane level with second messenger participation. Second, there would be gene activation; the pathways used by each cell type could provide specificity to the signal

Fig. 2. Integrated mechanism for testosterone effects in cardiomyocytes. In cardiomyocytes, testosterone activates plasma membrane androgen receptors coupled to heterotrimeric Gq protein. Gβγ dimers activate phospholipase C (PLC), which locally produces inositol IP3 thus activating IP3 receptors (IP3R). The consequent Ca2+ increase leads to the activation of several pathways, including the MEK/ERK/mTOR axis. These responses combined induce a series of genomic changes that lead to increased gene expression and increased protein

While studying these hypertrophy effects on cardiomyocytes, our laboratory determined that testosterone uses a cooperative mechanism between the genomic and nongenomic pathways to produce the hypertrophic growth of cardiac cells. Thus, with normal levels of androgens the interactions among signaling pathways lead to the maintained health of cardiac cells, whereas high doses recruit additional signals which are hypertrophic. As

synthesis, ultimately leading to cardiac hypertrophy.

and transcription factor activation, among other fast events.

through a cooperative mechanism, which can act in a concerted form.

depicted in Figure 2, testosterone activates a plasma membrane G-protein coupled receptor, a signaling pathway that results in a nongenomic mechanism. Activation of this pathway facilitates the hydrolysis of phosphatidylinositol 4,5-bisphosphate by phospholipase C, thereby generating diacylglycerol and IP3. IP3 mediates opening of IP3 receptors in the sarcoplasmatic reticulum, which in turn produce rapid (1-5 min) Ca2+ release (Vicencio et al., 2006). Similar effects of testosterone have been identified in T-cells, skeletal muscle and neuroblastoma cells (Benten et al., 1999; Estrada et al., 2003, 2006). In cardiomyocytes, nongenomic signals activate the extracellular signal-regulated kinase (ERK) and the mTOR pathways. Activation of Ca2+/ERK/mTOR axis is independent of androgen receptor. The anabolic properties of testosterone indicate that this hormone regulate multiple genes involved in growth and metabolism. Traditionally, ligand-dependent androgen receptors act on DNA within the nucleus and in this compartment there is cross-talk among the nuclear receptors and cytosolic signaling. We have determined that testosterone induce a hypertrophic pattern in cardiomyocytes, evaluated as increase in the expression of fetal proteins, cardiomyocyte size, and protein synthesis. Hypertrophy effects of testosterone were blocked by either inhibition of mTOR or androgen receptor (Altamirano et al., 2009). We propose that cell growth produced by androgens require both androgen receptor activity and translation control through mTOR signaling pathway. Thus, both mTOR pathway and androgen receptor could control protein synthesis by a coordinated mechanism, where mTOR regulates translation and the intracellular androgen receptor


**2.2 Case report Betty**

**Focus**: influence of pregnancy on HbA1c, INS/d, and BMI in a lady with T1D using a pump without CGM, and effects of hybrid MiniMed 780G implemented after pregnancy.

**History:** Betty (today 33 y/o) was diagnosed with T1D at the age of 14 years in September 2004 in pediatric department of regional hospital. She was immediately treated by means of an insulin pen with insulin aspart and detemir, a maximum dose of insulin was 20 IU/d. In 2007, insulin pump Animas was implemented as the main form of insulin therapy. In 2015 the pump Animas was replaced by MiniMed 640G. In the course of the first pregnancy, a decrease in HbA1c and marked increase in INS/d could be seen. She gave birth to a boy (delivery July 8, 2015 by Cesarean section, 4.48 kg, 54 cm). The parameters from August 31, 2015 demonstrated improved compensation (lower values of HbA1c), however, the amount of insulin per day was nearly doubled (**Table 2**, **Figures 5** and **6**). Betty had a second pregnancy in 2019 (delivery November 1, 2019 by Cesarean section, girl 4.75 kg, 53 cm). During this second pregnancy, there was an apparent decrease in HbA1c concentration and an increased supply of exogenous insulin (161 IU/d). To explain these similar changes in HbA1c and INS/d it is worth noting that placenta is permeable to glucose but impermeable to insulin [23, 24]. Therefore, the insulin from fetal pancreas may create new fetal adipose tissue by converting the glucose crossing through placental barrier from mother's plasma. These pathways may result in fetal macrosomia (which is a known complication associated with raised HbA1c concentration in the last trimester of pregnancy). Only a minority of pregnant women were able to optimize their glycemic control. This makes CGM and hybrid insulin pumps all the more favorable for treatment during pregnancy [25, 26]. However, Betty could receive the hybrid pump MiniMed 780G as late as in April 2021. Current TIR: 96% (January 5, 2023).

### **Table 2.**

*Betty's clinical and laboratory parameters over the last 8 years.*

*Benefits of Implementation of Insulin Pump in People with Type 1 Diabetes: 10 Case Reports DOI: http://dx.doi.org/10.5772/intechopen.110073*


*Edited by Antonio Vanderlei Dos Santos and Joao Carlos Krause*

The book presents a discussion on education of sciences, through a technological view shown in the works of a variety of authors from different countries. It's a differentiated conception of scientific education bringing renowned authors who discuss from teacher formation to the inclusion of new technologies into education. We are proud to say that the themes discussed in the book are up to date and also of scientific interest in many countries, as seen by the collaborating authors who come from many parts of the world. The scientific discussion becomes evident through the effort of the authors in participating in this book that will serve as a reference for future research for those who want to develop modern educational approaches.

ISBN 978-953-51-3539-5

Science Education - Research and New Technologies

Photo by ChristianChan / iStock


**1. Introduction**

A vast amount has been written over the last five decades on aggrecan's structure (Figure 1) and function in weight-bearing connective tissues such as hyaline cartilage and intervertebral disc (IVD) in health and disease [1–8]. However, aggrecan also has important roles in tensional connective tissues (e.g., meniscus, tendon and ligaments) [9,10] as well as in non-cartilaginous tissues such as the heart and nervous system [11–22]. While aggrecan has important roles in tissue development and function, surprisingly little has been published on its interactive and cell-directive properties in tissue

morphogenesis. This review aims to rectify this deficiency but cannot be meaningfully undertaken without first covering aggrecan's functional attributes in weight-bearing tissues that contribute to matrix stabilisation. This diversity in aggrecan's functional properties is due to modifications in its glycosaminoglycan (GAG) side chains which equip it with unique ligand interactivity in specific developmental contexts.

**Figure 1.** Amber/GLYCAM 3D stick structures of chondroitin sulphate isomers depicting their 3D conformations, disaccharide compositions and Haworth projection disaccharide structures showing sulphation positions (**a**–**e**). Schematic depiction of the structural organisation of the chondroitin sulphate glycosaminoglycan side chains of aggrecan depicting specific structural features of areas of the chain detected by monoclonal antibodies, putative sulphation patterns, linkage region structure to aggrecan core protein and non-reducing terminal structures (**f**). These regions on the CS side chain are numbered 1–8. Key: (1) Non-reducing terminal groups present on some cartilage aggrecan CS chains; (2) 3-B-3(−) CS sulphation motif is also present as a non-reducing terminal component on some chains; (3) putative region on CS chain identified by MAb 6C3; (4) putative region on CS chain identified by MAb 4C3; (5) putative region on CS chain identified by MAb 7D4; (6) CS linkage attachment region to Serine residues of the aggrecan core protein; (7) 3-B-3(+) CS sulphation stub epitope generated by exhaustive digestion of the CS chain by chondroitinase ABC and recognised by MAb 3-B-3; and (8) 2-B-6(+) CS sulphation stub epitope generated by exhaustive digestion of the CS chain by chondroitinase ABC and recognised by MAb 2-B-6. Note: Regions 3–5 of the CS chains containing the 6-C-3, 4-C-3 and

7-D-4 reactivity are susceptible to chondroitinase ABC digestion; thus, in graded partial digestions, the 6-C-3 and 4-C-3 reactivity can be selectively removed leaving the 7-D-4 reactive region intact. However, this is also susceptible to chondroitinase ABC, and exhaustive digestion conditions eventually lead to generation of the unsaturated 3-B-3(+) and 2-B-6(+) stub epitopes attached to the linkage region, as shown in this diagram. In (**f**), the structures shown hypothetical many features such as the sulphation positions on GAGs are variable; the depictions shown are thus generalisations based on literature data.


*3.4. Application of SemiTMC-GMM*

The question is now how to apply the proposed model to recognise lower limb locomotion activities. In our previous work [7], gait cycle was introduced into the estimation of four locomotion activities, and the results show that it can improve the accuracy. As introduced in [29], one gait cycle can be divided into four gait phases, i.e., stance, push-up, swing, and step down. In this work, we are pursuing a method that does not require the sensor to be placed on the feet only. On the contrary, it can be placed on different places of the lower limb, such as thigh, shank, or foot. The segmentation of gait cycle is based on the motion of foot, so similarly we can define 'leg cycle' based on the motion of leg. One leg cycle can be segmented into four leg phases, which are low position, lifting, high position, and dropping.

Let assume the hidden state *X* represents the activity, and *U* be the gait cycle or leg cycle. Thus, the dimension of Λ (*r*) depends on the number of activities; while for Γ, *τ* is equal to 4. The transition of *X* and *U* follows a specific order because the feet move from attaching on the ground to swinging in the air alternately, or the legs switch between lifting to dropping. Therefore, we define a specific transition graph for *X* and *U*. As shown in Figure 2, the numbers 1–4 represent the hidden state *U*, the four gait and leg phases. We can see that *U* transfers from phase 1 to phase 4 and back to phase 1 again cyclically if the activity does not change. While the activity is switching, *U* transfers from phase 1 of the previous activity to phase 2 of the current activity.

**Figure 2.** Hidden state transition graph. The activities represent *X*, and the numbers 1–4 represent *U* and stand for the four gait phases, or leg phases.

The hidden states *H* and *D* are not the final goals of the recognition, and they have no physical meaning neither. For simplification, the dimension of *L* (-) is set to 9. This value was determined by our experience, a too small value will make the results of SemiTMC-GMM no difference from that of TMC-GMM, while a too large value will cost too much time for running the code. The performance of different GMM components number (*κ*) is evaluated on two datasets, as depicted in Section 5.

The observation *Y* is the feature extracted from the sensor readings. The utilized features are the sliding mean value and standard deviation, by calculating the mean value and standard deviation of acceleration and angular rate within a sliding window length. Since IMUs measure 3-dimensional acceleration and angular rate, then the dimension of the observation *Y* (*w*) equals to 12. The initialization of the hidden states is the same as the one in our previous work [7], so it will not be repeated here. Afterward, based on the initial hidden states *vN* 1 and features *yN* 1 , the initial conditioned GMM density *p*(*yn*|*vn*, *hn*) can be easily obtained if the mixture number *κ* is known. When the initialization is done, batch mode EM algorithm can be applied to train the model, which will be described in details in Section 4. Then, the trained model can be used for the batch mode testing, or, as the initial model of on-line EM algorithm.


**4. Contribution of the electron transfer mechanism in photosensitized reaction by cationic porphyrins**

Photooxidation activity through electron transfer depends on the redox potential. It has been demonstrated that photoexcited hematoporphyrin, a free base porphyrin, induces the oxidative electron transfer from the tryptophan residue of bovine serum albumin [67, 68]. Cationic porphyrins show relatively small *E*red values due to their positive charge. In this section, several examples of electron transfer-mediated oxidation of biomolecules by cationic porphyrins.

#### **Figure 6.**

*Structures of H2TMPyP and ZnTMPyP (A), their binding interaction with DNA (B), and the electron transfer reactions (C). ABG: Amino benzoyl-L-glutamic acid.*

### **4.1 Protein photooxidation through electron transfer by cationic porphyrins**

The photosensitized protein damage by tetrakis(*N*-methyl-*p*-pyridinio) porphyrin (H2TMPyP, **Figure 6**) and its zinc complex (ZnTMPyP, **Figure 6**) was reported [69]. Photosensitized reaction of H2TMPyP has been extensively studied [14, 70]. Water-solubility of H2TMPyP and its analogues is appropriate for biological study. Furthermore, electrostatic interaction between these cationic porphyrins and biomacromolecules is considered to enhance the electron transfer reaction with targeting biomolecules. The ΦΔ value of H2TMPyP is relatively large [14, 69, 71], and photosensitized biomolecule damage caused by H2TMPyP through 1 O2 generation is generally accepted [70, 72]. However, *E*red of H2TMPyP is relatively small [27], and negative Δ*G* values for photosensitized oxidation of several amino acids through electron transfer are estimated. Therefore, electron transfer-mediated photooxidation of biomolecules is expected.

H2TMPyP and ZnTMPyP bound to HSA and caused photosensitized oxidation of the tryptophan residue [69]. Three amino acids–tryptophan, phenylalanine, and tyrosine–were also used as target biomolecules, and tryptophan and tyrosine were photodamaged by these cationic porphyrins. However, H2TMPyP and ZnTMPyP could not photosensitize the damage of phenylalanine. The protein damage (oxidation of the tryptophan residue) was enhanced in deuterium oxide and inhibited by NaN3. Analysis of the scavenger effect showed that the absolute quantum yields of electron transfer-mediated oxidation are 5.3 × 10−3 and 4.0 × 10−3 for H2TMPyP

#### *Electron Transfer-Supported Photodynamic Therapy DOI: http://dx.doi.org/10.5772/intechopen.94220*

and ZnTMPyP, respectively. The *E*red of H2TMPyP (−0.23 V vs. SCE) [27] is lower than that of ZnTMPyP (−0.85 V) [73]. The values of -Δ*G* for electron transfer from tryptophan to their S1 states suggest that H2TMPyP (−1.03 eV) is more oxidative than ZnTMPyP (−0.53 eV). The estimated value of *k*ET estimated from the fluorescence lifetime for H2TMPyP was 1.0 × 108 s−1. On the other hand, the fluorescence lifetime of ZnTMPyP was not affected by the interaction with HSA in the presented experimental condition. Because of the relatively shorter fluorescence lifetime of ZnTMPyP (1.3 ns), the estimation of *k*ET may be difficult by the fluorescence lifetime measurement. Furthermore, protein photodamage by the T1 states of H2TMPyP and ZnTMPyP were also discussed [69]. The lifetimes of their T1 states are relatively long: H2TMPyP (2.1 μs) and ZnTMPyP (2.7 μs), suggesting that the electron transfer in the T1 state is kinetically advantageous. The estimated -ΔG of the electron transfer from tryptophan to their T1 states (−0.65 eV for H2TMPyP and − 0.15 eV for ZnTMPyP) suggests that this electron transfer is also possible in terms of energy.