**Table 3.**

*Applications of alginate in the fields of agriculture.*

bacteria as plant growth stimulants. Due to the environmental dangers of chemical pesticides, alternative solutions, such as biological control agents have been proposed to solve this problem. One of these promising solutions is the use of insecticidal bacteria, such as *Bacillus thuringiensis* Berliner. For this purpose, the bacteria are placed in a biodegradable alginate coating and sprayed on the plants. After the insects use these capsules, the insects die due to the production of toxic substances by the bacteria [68].

Rhizosphere bacteria with beneficial effects on plant growth and development are known as plant growth-promoting bacteria (PGPR). This group of bacteria is able to affect plant growth in direct or indirect ways. The direct effect includes the production or release of secondary metabolites, such as regulators or growth hormones and, such substances or facilitating the absorption of nutrients from the plant's growth environment. But the indirect effect of PGPRs in stimulating plant growth occurs when the harmful effects of one or more pathogens are reduced or completely prevented. Also, these bacteria are used for bioremediation or to improve the efficiency of other bioremediation methods of soils contaminated with heavy metals. The use of alginate capsules has been attention due to characteristics, such as compatibility with the environment, low cost compared to physical and chemical methods, stabilization and stability during transportation, help to improve soil fertility, etc. These bacteria are inserted in the alginate capsule and during the transfer process, soil stresses, and factors, such as pH and soil moisture are maintained and perform their functions in the best way [69, 70]. Alginate capsules containing "*Pantoea agglomerans*" bacteria have been used as bio fertilizers compatible with the environment as well as controlling some pests [71].

One of the essential uses of alginate in agriculture is the use of encapsulation to transfer beneficial probiotic bacteria into the body of livestock. Transfer of probiotic bacteria into the alginate capsule perform to improve the conditions and survival of these important bacterial species. Food in the digestive system of livestock can be better digested and absorbed in the presence of probiotic bacteria. Prebiotics are food substances that are resistant to digestion in the upper part of the gastrointestinal tract and reach the lower part of the intestine intact. There, with their direct physiological effect, as well as by the specific stimulation of the growth and activity of the probiotic compounds of the gastrointestinal microflora, including Bifidobacterium, they affect the ecosystem of the gastrointestinal system. Alginate is used for the transfer and function of these materials. Alginate is oftentimes used as an ideal substrate for the delivery of probiotics as well as prebiotics due to its good biodegradability and high compatibility with the body, lack of toxicity (**Figure 4**) [23, 72].

Also, other advantages of using alginate include the slow release of enclosed microorganisms and beneficial substances, low cost, and resistance to stomach acids and enzymes. Another advantage of using alginate is its combination with other useful compounds for the transmission and control of diseases in plants and animals in the agricultural sector. Studies have shown that alginate with chitosan creates a more stable and compatible combination. Also, the controlled release of compounds that are transported, such as various chemical or biological agents, is possible using the combination of alginate and chitosan. A clear example of this is the transfer of beneficial probiotic bacteria, such as Bifidobacterium and Lactobacillus into the livestock body. By combining alginate with gelatin, microcapsules can be produced, which are used to transport some useful bacteria, such as *Pseudomonas fluorescens*, and this increases the growth rate of potato plants. Also, this bacterium is used to control some diseases, such as dry rot in potatoes [23, 72].

Application of seed strengthening methods in order to eliminate or reduce environmental stress, increase germination percentage and seed yield is of high value. In some studies, it has been shown that the effect of different concentrations of alginate *Applications of Alginate in the Fields of Research Medicine, Industry and Agriculture DOI: http://dx.doi.org/10.5772/intechopen.110209*

**Figure 4.** *Using alginate microcapsules to transfer useful bacteria and micronutrients to the plant.*

biopolymer as a seed coating is effective on the physiological stages of corn seedling growth under the stress of oil pollution [73].


**4. Interaction of the zero-modes with a variable -term**

14 Quantum Gravity

is invariant in time. From this assumption one can prove in a straightforward way several strange properties of particles with negative mass. These properties can be summarized by saying that in the usual dynamical rules their mass really behaves like a negative number, namely: (a) The acceleration of the virtual particle is opposite to the applied force. (b) The momentum is opposite to the velocity. (c) The kinetic energy is negative. The kinetic energy is defined as usual through the work of the applied force, in such a way that the sum

Applying these rules one obtains a bizarre behaviour in the scattering processes and in the dynamics. For instance, although the gravitational potential energy of two virtual particles with negative mass is negative, *E*pot =-*GM*1*M*2/*r* (compare Sect. 3.1), the two particles experience a repulsion, due to Property (a). They tend to run away from each other; while their distance increases, their *E*pot decreases in absolute value, and their (negative) *E*kin increases in absolute value. If the particles were initially at rest at some distance *r*0 (Fig. 5),

when their distance goes to infinity they gain a *E*kin equal to their initial *E*pot.

Fig. 5. "Classical" motion of two virtual particles with negative mass initially at rest at distance *r*0. Although their potential energy is negative, they feel a repulsion and their (negative) kinetic energy increases in absolute value as their distance goes to infinity.

recoil of the zero-modes (in the same direction of the emission). The conservation equations

2 0 *r g r g MvE E Mv p*

where *E* is the energy gap, *E*g and *p*g are the graviton energy and momentum, *v*r is the recoil velocity of the zero-mode and 2*M*-10-13 kg is the zero-mode mass. After replacing

1 0 *EE E g g M*

2

2 2

(Sect. 3.2) the momentum of the emitted graviton is balanced by the

(27)

(26)

*E*kin+*E*pot is conserved.

In the decay

give

 *p*g=*E*g, the system (26) leads to the equation

In Sect. 3 we have computed the probability of the decay process with emission of a virtual graviton. The excitation process (transition of a zero-modes pair from a symmetric to an anti-symmetric state) can occur by absorption of a virtual graviton or by coupling to an external source. It is easy to show (Sect. 4.3) that the coupling of zero-modes

to "ordinary" matter with energy-momentum *dx dx T m d d* is exceedingly weak.

(Note that certain interactions between zero-modes and massive particles vanish exactly for symmetry reasons. For instance, a particle in uniform motion can never "lose energy in collisions with the zero-modes", because in its rest reference system the particle will see the vacuum, zero-modes included, as homogeneous and isotropic. There are possible exceptions to this argument: accelerated particles, or particles in states with large *p* uncertainty.)

The coupling to a (*t*) term, or local time-dependent vacuum energy density, can lead to a significant transition probability. This is due to the presence of the non-linear *g* factor in the coupling, and corresponds physically to the fact that such a term does not describe isolated particles, but coherent, delocalized matter.


**Emerging Micro Manufacturing Technologies and Applications**

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**Nikolaos Tapoglou**

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This is a reprint of articles from the Special Issue published online in the open access journal *Micromachines* (ISSN 2072-666X) (available at: www.mdpi.com/journal/micromachines/special issues/micro manufacturing technologies).

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*4.4. Protein Expression, Purification, and ATPase Assay*

The *OsAAA-ATPase1* sequence was amplified by PCR and cloned into the sites between BglI and HidIIIp of pET32a (Novagen) as a His-tag fusion protein; then, that was transfected into the *Escherichia coli* Origami strain BL21(Lys). The set of primers used was as follows: OsAAA1BglII 5- -GTAGATCTCTTGAGACAAATGGAGGCGACG-3- ; OsAAA1HindIII 5- -GCTAAGCTTCTACTTATCCTTCCCGACCAC-3- . Expression of the protein was induced for 4 h at 25 ◦C with 0.5 mmol/L isopropyl β-d-1-thiogalactopyranoside. *Escherichia coli* cells were pelleted by centrifugation, resuspended in lysis buffer (20 mmol/L Tris-HCl pH 7.4, 0.1 M NaCl, 10 mmol/L imidazole), and sonicated. After the cell debris was removed by centrifugation (12,000 × g, 10 min, 4 ◦C), the supernatant was loaded onto a High Affinity Ni-Charged Resin (GE Healthcare, Buckinghamshire, UK), washed with washing buffer (20 mmol/L Tris-HCl pH 7.4, 0.1 M NaCl, 10 mmol/L imidazole), and eluted with elution buffer (20 mmol/L Tris-HCl pH 7.4, 0.1 M NaCl, 180 mmol/L imidazole). ATPase activity was measured by the malachite green-based colorimetric method using the QuantiChromTM ATPase/GTPase activity assay kit (Sigma-Aldrich, St. Louis, MO, USA). The elution buffer was used as the negative control, and an ATPase from potatoes (Sigma-Aldrich, St. Louis, MO, USA) was used as the positive control. One unit is defined as the amount of enzyme that catalyzes the production of 1 μM of free phosphate per minute under the assay conditions.

#### *4.5. Subcellular Localization*

For subcellular localization of OsAAA-ATPase1, the plasmid pSAT6-AFP-C1-OsAAA1 was transformed into protoplasts prepared from etiolated seedlings as previously described [52]. As a control for cytoplasmic localization, the pSAT-mCherry construct was co-transformed. Fluorescence was examined under a confocal microscope (Leica Microsystems, Wetzlar, Germany) 16 h after transformation.


**Novel Tissue Types for the Development of Genomic Biomarkers**

Zinaida Sergueeva, Heather Collins, Sally Dow, Mollie McWhorter and Mark L. Parrish *Covance Genomics Laboratory, Seattle, Washington, USA*