**Author details**

Raymond A. Cloyd Kansas State University, Manhattan, KS, USA

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© 2019 The Author(s). Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/ by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

**7**

*Effects of Pesticides and Adjuvants on the Honey Bee,* Apis mellifera*: An Updated Bibliographic…*

applied to California almond orchards

[10] Greig-Smith PW, Thompson HM, Hardy AR, Bew MH, Findlay E, Stevenson JH. Incidents of poisoning of honey bees (*Apis mellifera*) by agricultural pesticides in Great Britain 1981-1991. Crop Protection.

[11] Cloyd RA. Pesticides and bees. Kansas State University Agricultural Experiment Station and Cooperative Extension Service. MF3428. Manhattan, KS: Kansas State University; 2018

[12] Atkins EL, Kellum D. Comparative morphogenic and toxicity studies on the effect of pesticides on honeybee brood. Journal of Apicultural Research.

Delpuech J-M. The sublethal effects of pesticides on beneficial arthropods. Annual Review of Entomology.

[14] Zhu W, Schmehl DR, Mullin CA, Frazier JL. Four common pesticides, their mixtures and a formulation solvent in the hive environment have high oral toxicity to honey bee larvae. PLoS One.

[15] Needham PH, Stevenson JH. The toxicity to foraging honeybees, *Apis mellifera*, of endosulfan, malathion and azinphos-methyl applied to flowering oil seed rape, *Brassica napus*. Annals of Applied Biology. 1973;**75**:235-240

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and Behavior. 2005;**82**:30-39

*mellifera*). Pharmacology Biochemistry

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insects10010020

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*DOI: http://dx.doi.org/10.5772/intechopen.89082*

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[3] Lautenbach S, Seppelt R, Liebscher J, Dormann CF. Spatial and temporal trends in global pollination benefit. PLoS One. 2012;**7**:1-16. DOI: 10.1371/

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[5] Ciarlo TJ, Mullin CA, Frazier JL, Schmehl DR. Learning impairment in honey bees caused by agricultural spray adjuvants. PLoS One. 2012;**7**:e40848

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[9] Wade A, Lin C-H, Kurkul C, Regan ER, Johnson RM. Combined toxicity of insecticides and fungicides *Effects of Pesticides and Adjuvants on the Honey Bee,* Apis mellifera*: An Updated Bibliographic… DOI: http://dx.doi.org/10.5772/intechopen.89082*


**4. Glutathione system and multiple sclerosis**

**Figure 4.** Effect of nitric oxide in inflammation. Pathway Builder Online Tool was used to draw the figure [27].

chain by nitric oxide, which in turn causes a decrease in Na+

+

glutamate antagonist [10].

154 Trending Topics in Multiple Sclerosis

Paraclinical studies have shown an increased metabolism of the RNS in serum, lymphocytes, and cerebrospinal fluid of MS patients, which correlate to pathology studies. ONOO-

closely associated with acute inflammatory lesions [9]. Damage to axons is mediated by the following: (1) failure in mitochondrial energy metabolism due to inhibition of the respiratory

‐dependent glutamate transporters, (2) over‐expression of glutamate receptors, (3) oligoden‐ droglial excitotoxicity, (4) massive influx of extracellular Ca++, (5) activation of proteases, and (6) impaired axonal transport. These mechanisms produce glutamate excitotoxicity and increased generation of nitric oxide leading to nitrosative stress.  When nitric oxide is combined with the superoxide anion, it generates a potent free radical, the pro‐oxidant peroxynitrite. Glutamate in turn causes neurodegeneration through the AMPA and NMDA receptors in oligodendrocytes and astrocytes (**Figure 3**). It is possible to explain the role of mediators using an experimental model of autoimmune encephalitis: Protection against the experimental disease occurs after administration of a

Under physiologic conditions, nitric oxide is produced from L‐arginine by constitutive nitric oxide synthase (cNOS) and participates in a variety of important biological functions such as immunoregulation of inflammatory reactions, the downregulation of tumor necrosis factor (TNF)‐α production, MHC II expression in macrophages, induction of apoptosis in CD4 cells, physiological regulation of the mitochondrial respiratory chain, inhibition of antigen presen‐ tation, and leukocyte adhesion and migration. However, during inflammatory reactions, exposure of macrophages to interferon (IFN)‐γ and TNF‐α results in the activation of the

is also

/K+ ATPase activity and alters Na

In a recent study, the oxidation of DNA in the nucleus of oligodendrocytes and oxidation of lipids in the myelin of oligodendrocytes and axons were observed. This oxidation was associated with the active process of demyelination and neurodegeneration. Active lesions in relapsing‐remitting MS (RRMS) and progressive course patients were associated with inflammation, lipid peroxidation, and DNA oxidation [11]. Similarly, Ortiz et al. [12] observed an increase in serum lipid peroxides and nitrite/nitrate levels and the activity of glutathione peroxidase in patients with RRMS compared to healthy individuals.

Reduced ubiquinone and vitamin E levels, and reduced activity of the enzyme glutathione peroxidase in lymphocytes and granulocytes were reported (with a decrease in 51 and 78%, respectively), as well as a decrease in glutathione reductase activity in granulocytes (27%) and lymphocytes (8%) [13]. In contrast, in 2012, Tasset et al. [14] found an increase in activity of the glutathione reductase in patients with RRMS when compared to control subjects (1.3 ± 0.9 vs 0.3 ± 0.19, P < 0.01), and an increased ratio of reduced glutathione to oxidized glutathione (GSH/GSSG) in these patients (28.2 ± 39.6 vs 4.0 ± 2.9, P < 0.01). Similarly, an increase was found in the levels of oxidized glutathione and also increased concentrations of isoprostanes and malondialdehyde (MDA) in patients with MS [15, 16].

#### **4.1. Glutathione deficiency and multiple sclerosis**

There are several reports in the literature that relate the decrease or alteration of glutathione (GSH) metabolism with several neurodegenerative diseases. Biochemical analysis of postmor‐ tem brains has provided evidence for the generation of oxidative stress during the course of the disease since the total GSH content is reduced by 40–50% compared to controls. Also in several brain regions, we have found increased levels of lipid peroxidation [17]. The ratio GSH/ GSSG (usually 10:1) is considered consistent with the concept of oxidative stress as an important part in the pathogenesis of MS. Moreover, low concentrations of GSH appear to be an important indicator of oxidative stress during the progression of MS. Although the decrease in GSH alone is not responsible for the degeneration of glial cells and neurons, reduced GSH could increase the susceptibility to other stressful factors and contribute to neuronal damage at glia and neuron cells. Glutathione has been reported to protect mitochondrial complex I activity against nitrosative stress, as S‐nitrosoglutathione is formed. When this complex increases its content of nitrotyrosine and nitrosothiol groups in response to nitrosative stress, its activity is inhibited and therefore ATP production is diminished, which causes neuronal degeneration [10]. The role of glial cells in generating ROS in MS and the selective vulnerability of neurons is due to activated glial cells surrounding these neurons, as these glial cells are also directly involved in GSH levels. The engagement of the glutathione system in astroglial cells contributes to the reduction in its antioxidant defenses and so poor glial defense could contribute to existing neuronal damage (**Figure 5**) [10]. Furthermore, the specific activities of some enzymes that metabolize GSH are high, as in the case of glutathione peroxidase, glutathione reductase, and glutathione S‐transferase. Other products of OS are also elevated, as in the case of 4‐hydroxynonenal (4‐HNE, a product of lipid peroxidation of polyunsaturated omega‐6 fatty acids) [17].

**Figure 5.** Genetic defect in glutathione synthesis and neurodegenerative diseases. Pathway Builder Online Tool was used to draw the figure [27].

A new proposal is that a genetic defect of glutathione synthesis may be the initial event in the failure of the antioxidant defenses. In neurodegenerative diseases, a decreased GSH level is accompanied by dysfunction of the mitochondrial complex I and complex IV and promotes oxidative stress [18]. We found a significant decrease in GSH levels in the cerebrospinal fluid of patients with this disease, and, in addition, proton magnetic resonance studies have shown a 50% decrease in GSH levels in the frontal cortex of patients with MS (**Figure 5**).


**2.4 Drop volume/weight method**

50 Toxicity and Drug Testing

made up of platinum or iridium is submerged into liquid and then pulled through the liquid-air interface. Maximum pull exerted on the ring by the surface is measured which is directly proportional to the surface tension value at equilibrium (Bodour and Miller-Maier, 1998). The ring is submerged into the solution and then slowly pulled through the liquid-air interface to detach from the interface with a force that is correlated to the surface tension. With this method, it is possible to measure the interfacial tension at both liquid-air and

> cos *F f*

where *p* is the perimeter of the three-phase contact line, *f* is the correction factor (because additional volume of liquid is lifted during the detachment of the ring from the interface) between each measurement. The platinum wire ring was rinsed three times with water, later

One major difference between the Du Noüy ring method and Wilhelmy's plate is the way in which the surface tension measurement is carried out. The ring moves through the interface whereas the plate is static at the interface, therefore there is no disturbance at the interface and this method is the recommended geometry for studying time dependent characteristics. Both ring and plate geometries can be used with the force balance type of tensiometer. A single instrument is normally capable of performing either Wilherlmy plate or du Noüy ring

The surface tension measured by du Noüy method has been utilized in pharmaceutical research, for example, in the measurement of emulsion stability (Ishii et al., 1988; Takamura et al., 1984) and development of a dissolution media to simulate the physiological environment of the gastric region (Luner et al., 2001). This method can be used for characterization of pharmaceutical formulations such as plasticizer and polymer coating and surface tension calculation of various surfactant solutions and their CMC values (Palma et al., 2002) at the point of intersection of the interfacial tension value versus surfactant

The maximum bubble pressure method involves flow of a gas bubble (typically air or nitrogen) at a constant rate and blows them through a capillary with a known diameter which is submerged in the sample liquid. The pressure inside of the gas bubble increases until the bubble becomes hemispherical and its radius corresponds to the radius of the capillary. Beyond this the bubble is unstable and grows explosively until it detaches itself from the capillary and a new bubble is formed. The method is based on the continuous measurement of the applied pressure versus bubble rate formed at the end of the capillary. Figure 1 shows each step of bubble formation and corresponding change of bubble radius.

The dynamic surface tension can be directly calculated by Young-LaPlace equation:

max 2 *P R*

where Δ*P*max is the maximum pressure difference and *R* is the capillary radius (Drelich et al., 2002; Hallowell and Hirt, 1994; Fainerman et al., 2006). This method is one of the most popular

(3)

concentration plot (de la Maza, 1998; Korhonen et al., 2004; Zelkó et al., 2002).

(2)

*p*

liquid-liquid interfaces. Surface tension can be calculated using the equation below:

with acetone as was blow dried (Drelich et al., 2002).

measurements (Thiessen and Man, 1999).

**2.3 Maximum bubble pressure method**

Among the conventional methods of surface tension measurement, drop shape techniques have proven to be reliable and easy to handle. This method weighs the mass of the liquid drop or the volume of the drop that falls off a capillary tip of known diameter when pumped very slowly. The weight of the drop falling off the capillary correlates with the interfacial tension and is measured by balancing it against a known gravitational force through the following equation:

$$\mathcal{W} = V \Delta \rho \text{g} = 2 \pi r f \tag{4}$$

where is the difference in the density of the heavy phase and the light phase, *g* is the gravitational constant (*g*=9.81652 m/s2), *r* is the radius of the capillary tip and *f* denotes the empirical drop correction factor introduced by Harkins and Brown. The correction factor is required because only a portion of the drop falls from the capillary tip during detachment and this corrects the deviation of the drop volume from its ideal value (Drelich et al., 2002; Gunde et al., 2001). Impurity of active pharmaceutical solutions (Al-Maaieh and Aburub 2007), emulsion stability (Rangsansarid and Fukada, 2007), potency of local anesthetics (Matsuki et al., 1998), stability of biphasic aqueous systems (Mishima et al., 1998) and surface active properties of drugs (Deo et al., 2004) have been evaluated using this technique.


*2.1. PV Models*

SDM, DDM, and TDM models have been widely used by researchers in recent years [20]. In general, more diodes in a circuit represent a more accurate model, but also increase the model complexity [33].


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