**1. Introduction**

3

Chicken meat is the most accessible protein source for humans in most countries. However, the poultry sector may be limited by a series of problems, including high or low environmental temperature, high stocking density, etc. Heat stress is one of the main problems encountered within the poultry sector [1,2]. Heat stress affects nutritional quality, resulting in a lower protein content and higher fat deposition, etc. [3], and sensorial quality such as low water-holding capacity, higher brightness, and lower redness of meat, etc. [4]. In recent years, much attention has been paid to the inclusion of phytochemicals (vitamins, minerals, phytogenic feed additives) with antioxidant properties in the feed mixture, which would overcome the effects of heat stress on poultry health and on the quality of the meat they produce [5]. Some authors have stated that dietary Cr supplementation had a positive effect on meat quality by decreasing the fat content of the carcass, [6] and on carcass traits of broiler chicks in natural [7,8] or heat stress conditions [9]. Moreover, it has been suggested that combinations between Cr and other antioxidants (Zn, vitamin C) might have a synergistic action [2,10].

Vitamin C is a renowned water-soluble vitamin, with undeniable antioxidant activity. In heat-stressed broilers, dietary vitamin C supplementations have improved performance and humoral immunity [11].

Zinc is one of the most important components of the poultry diet, being a cofactor for enzymes with implication in the antioxidant defense system [12]. It was reported

**Citation:** Saracila, M.; Panaite, T.D.; Mironeasa, S.; Untea, A.E. Dietary Supplementation of Some Antioxidants as Attenuators of Heat Stress on Chicken Meat Characteristics. *Agriculture* **2021**, *11*, 638. https://doi.org/10.3390/ agriculture11070638

Academic Editors: Lubomira Gresakova and Emilio Sabia

Received: 28 May 2021 Accepted: 7 July 2021 Published: 8 July 2021

**Publisher's Note:** MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations.

**Copyright:** © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/ 4.0/).

that dietary Zn supplementation in broiler chickens improved broiler carcass quality by reducing the percentage of abdominal fat, suppressing lipid peroxidation of chicken meat [13].

Creeping wood sorrel (*Oxalis corniculata*) is a little-known plant, which contains an important amount of vitamin C [14] and phenolic compounds with strong free radical scavenging activity [15]. Previous studies demonstrated that including creeping wood sorrel in chicken diet (1%) and Cr picolinate (0.2 mg/kg diet) could counteract the effects of heat stress, resulting in unaffecting growth performance, improving lactobacilli populations and reducing pathogenic bacteria [14].

However, dietary supplementation with Cr together with other minerals such as Zn, vitamins (C, E), or phytogenic feed additives may potentate the antioxidant effect. Therefore, the objective of this study was to assess the effect of dietary inclusion of chromium and vitamin C, Zinc, and sorrel wood powder supplements on chicken health and nutritional, textural, and sensorial quality of chicken meat.

#### **2. Materials and Methods**

#### *2.1. Birds, Diets, and Treatments*

A 6 week feeding trial (0–42 d) was conducted on 120 unsexed Cobb 500 chickens (1 day of age), weighed and randomly divided into four homogenous groups (C, E1, E2, and E3) with 30 chickens/group. Each group consisted of 6 replicates, with 5 chickens per replicate (1 replicate/chickens/cage). The feeding trial was conducted in an experimental hall of the National Institute for Animal Nutrition (Ilfov, Romania) according to experimental protocol approved (case no. 4775/02.08.2019) by the Ethics Commission of the Institute (Ethical Committee no. 52/30.07.2014). The chicks were housed in three-tiered digestibility cages (cage dimensions 65 × 75 × 45 cm, one cage per replicate) During the experimental period, the chickens were reared under controlled environmental conditions and monitored by a Viper Touch computer (temperature 32 ± 0.5 ◦C, humidity 36 ± 1.4%, with 0.38 ± 0.01% ventilation/broiler, and 899 ± 0.2 ppm CO2 emission). The light regimen was 23 h light/1 h darkness. Compared with the control diet (C group), the experimental diets (E1, E2, and E3 group) included 200 μg/kg diet chromium picolinate, CrPic (Table 1). Additionally, the experimental diets contained 0.25 g vitamin C (VC)/kg diet (E1), 0.025 g Zn/kg diet (E2), and 10 g creeping wood sorrel powder (CWS)/kg diet (E3). The chickens had free access to feed and water. Creeping wood sorrel was analyzed, and the data was published previously by Saracila et al. [14]. The analysis revealed a concentration of 15.44% CP, 2.83% EE, 4.96 mg/g GAE total polyphenols, 31.60 mmol ascorbic acid equivalent antioxidant capacity and 11.77 mg/100 g vitamin C.

#### *2.2. Sample Collection and Analysis*

At the end of experiment (42 days), from each group with homogenous weights, 6 chickens/group were randomly selected (1 chicken/replicate). From each group selected, blood samples were collected aseptically from the sub-axial region into heparinized test tubes. The blood samples were centrifuged (775× *g* for 25 min at 4 ◦C), and the serum obtained was analyzed using an automatic BS-130 chemistry analyzer (Bio-Medical Electronics Co., Ltd., Shenzhen, China) in order to determine the biochemical parameters (serum glucose, cholesterol, triglycerides, phosphorus, calcium, iron, alanine aminotransferase, and aspartate aminotransferase).


*Article* **In Vitro Toxicological Profile of Labetalol-Folic Acid/Folate Co-Administration in H9c2(2-1) and HepaRG Cells**

**Robert Rednic 1, Iasmina Marcovici 2,3, Razvan Dragoi 1,\*, Iulia Pinzaru 2,3,\*, Cristina Adriana Dehelean 2,3, Mirela Tomescu 1, Diana Aurora Arnautu 1, Marius Craina 1, Adrian Gluhovschi 1, Mihaela Valcovici 1 and Aniko Manea 1**

**Abstract:** *Background and Objectives:* The consumption of dietary supplements has increased over the last decades among pregnant women, becoming an efficient resource of micronutrients able to satisfy their nutritional needs during pregnancy. Furthermore, gestational drug administration might be necessary to treat several pregnancy complications such as hypertension. Folic acid (FA) and folate (FT) supplementation is highly recommended by clinicians during pregnancy, especially for preventing neural tube birth defects, while labetalol (LB) is a β-blocker commonly administered as a safe option for the treatment of pregnancy-related hypertension. Currently, the possible toxicity resulting from the co-administration of FA/FT and LB has not been fully evaluated. In light of these considerations, the current study was aimed at investigating the possible in vitro cardio- and hepatotoxicity of LB-FA and LB-FT associations. *Materials and Methods:* Five different concentrations of LB, FA, FT, and their combination were used in myoblasts and hepatocytes in order to assess cell viability, cell morphology, and wound regeneration. *Results:* The results indicate no significant alterations in terms of cell viability and morphology in myoblasts (H9c2(2-1)) and hepatocytes (HepaRG) following a 72-h treatment, apart from a decrease in the percentage of viable H9c2(2-1) cells (~67%) treated with LB 150 nM–FT 50 nM. Additionally, LB (50 and 150 nM)–FA (0.2 nM) exerted an efficient wound regenerating potential in H9c2(2-1) myoblasts (wound healing rates were >80%, compared to the control at 66%), while LB-FT (at all tested concentrations) induced no significant impairment to their migration. *Conclusions:* Overall, our findings indicate that LB-FA and LB-FT combinations lack cytotoxicity in vitro. Moreover, beneficial effects were noticed on H9c2(2-1) cell viability and migration from LB-FA/FT administration, which should be further explored.

**Keywords:** labetalol; folic acid; folate; dietary supplements; cytotoxicity


**Acknowledgements**

The authors wish to thank PetroChina Tarim Oil Company for providing us with the data and for permission to publish this paper, and Baker Hughes internal support to carry out the work.


**Author details**

Opeyemi U. Lawal, Valeria R. Parreira and Lawrence Goodridge\* Canadian Research Institute for Food Safety, Department of Food Science, University of Guelph, Ontario, Canada

\*Address all correspondence to<EMAIL_ADDRESS>
© 2022 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.


**1. Introduction**

168 Gastrointestinal Endoscopy

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The diagnosis of glandular atrophy of the gastric mucosa (gastric atrophy) remains a challenge in the areas of gastrointestinal endoscopy and pathology. The importance of an endoscopic suspicion of this regressive tissue change is the possibility of alerting the pathologist to its presence. Patients with gastric atrophy, especially in more advanced stages, are more prone to develop intestinal metaplasia, dysplasia and gastric carcinoma, known as the Correa cascade (Correa, 1984, 1992). The most common type of gastric atrophy is that associated with *Helicobacter pylori* (*H. pylori*) infection. In such cases, the glandular atrophy usually occurs in parallel to the course of the inflammatory process that takes place in the gastric antrum (antral gastritis, multifocal gastritis) or in both gastric antrum and body (pangastritis, multifocal gastritis). In contrast, in most cases the chronic atrophic gastritis selective of the gastric body and sparing the antral mucosa is a consequence of an autoimmune inflammatory process. For the purpose of this chapter, the chronic gastritis presenting this histopathological pattern will be called atrophic body gastritis (ABG). Exceptionally, some patients with *H. pylori*-associated gastritis develop gastric lesions with histological pattern very similar to that of ABG which can lead to uncertainty about the differential histologic diagnosis with chronic gastritis of autoimmune origin.

Therefore, the two most important inflammatory diseases of the gastric mucosa, multifocal chronic gastritis and autoimmune gastritis, tend to progress to glandular atrophy of the gastric mucosa. In the first case, gastric atrophy may not occur or it develops more slowly, becoming conspicuous usually in later stages of life. In the second case, which is also more frequent with advancing age, gastric atrophy progresses more rapidly and may induce severe gastric changes also in the younger age groups. This differential course of the development of gastric atrophy in these two inflammatory diseases of the stomach involves different clinical and pathophysiological consequences. Those associated with *H. pylori* infection usually do not involve important pathophysiological changes, while the atrophy resulting from autoimmune disease often leads to well-known pathophysiological changes of the gastric mucosa, often culminating in pernicious anemia. Furthermore, the *H. pylori*dependent glandular atrophy is considered to be a condition predisposing to gastric adenocarcinoma, which has high rates of morbidity and mortality, while the autoimmune-

Atrophic Body Gastritis: A Challenge for the

correlated with the endoscopic reports (Table 2).

and gender

Presumptive Endoscopic and Histologic Diagnosis of Autoimmune Gastritis 171

(Table 1). The 196 patients with a final histologic diagnosis of ABG ranged in age from 11 to 94 years, with a significant predominance of females (76.0%) over males (24.0%). Fifty patients were in a relatively young age range (31 to 50 years), with an even more significant predominance of females, i. e., 83.3% *vs* 16.7% (Figure 1) .The remaining 175 patients were excluded from the study, either because the biopsy specimens of the antral or oxyntic mucosa were not fairly representative for histology or because they could be considered as cases of multifocal atrophic gastritis, regardless of the presence of *H. pylori* infection. Finally, the histological findings of the 196 patients with a diagnosis of atrophic body gastritis were

> All cases of chronic gastritis 2.564 100 Atrophic body gastritis 141 5,5 Unspecified atrophic gastritis 230 9,0

Total of ABG diagnosed 196 7,6

ABG after review of unspecified atrophic gastritis 55 2,2

Table 1. Main histological diagnosis of 6,005 gastroesophageal endoscopies carried out from 2007-2009 in a tertiary care unit of gastrointestinal endoscopy, Belo Horizonte, Brazil

Fig. 1. Distribution of the 196 patients with the final diagnosis of ABG according to the age

Among the 2,564 patients with chronic gastritis, 196 (7.6%) had a conclusive histologic diagnosis of atrophic body gastritis (type A chronic gastritis). Gastric mucosa samples of almost all of these patients showed well preserved antral mucosa, and severe glandular

**Total of cases %**

dependent glandular atrophy is considered to be a condition predisposing to gastric carcinoids, which, unlike the adenocarcinomas, have low rates of morbidity and mortality. Due to the different regional involvement of the stomach by these two types of chronic gastritis, their histological recognition is relatively easy when tissue samples are properly collected and processed. However, this is not always the case.

Thus, the importance of the differential diagnosis of ABG within this group of chronic gastritis resides in the possibility of its autoimmune origin and in its major clinical consequence, i.e., pernicious anemia. Pernicious anemia and autoimmune gastritis progress in an insidious manner and are usually diagnosed when they have reached florid clinical and morphological characteristics after years of evolution. However, a number of patients with dyspeptic complaints and patients with ABG and pernicious anemia undergo endoscopy in tertiary care settings without a prior knowledge of major illness. (Carmel, R., 1996) Therefore, for some patients seeking a first endoscopy service, the collection of biopsies from the gastric mucosa may be the first opportunity to establish the initial diagnosis of ABG, with or without pernicious anemia. This is important because the evolution of subclinical gastric lesions and systemic symptoms may result in undesirable consequences that become established in a gradual manner. These consequences mainly result from achlorhydria, vitamin B12 deficiency, and the development of hyperplastic polyps and neuroendocrine tumors of the gastric mucosa. For this reason, the diagnosis of ABG acquires importance since it establishes an appropriate clinical and endoscopic followup of the patient.

The objective of this article is to present some data on this topic obtained from a tertiary care unit of gastrointestinal endoscopy and to discuss some pitfalls linked to the routine histopathologic diagnosis of atrophic body gastritis.


*2.2. Synthesis Methods for Weld Lake Pigments*

Research on the W&N 19th century Archive Database was carried out under the sub-topic weld, which resulted in 12 database records. Among these, it was possible to identify seven records for the production of weld lake pigments, however, there are only five recipes as 3 of the records are copies, see Table 1. The remaining five database records include two notes on weld, two experiments to extract weld, and one recipe to prepare Yellow Carmine from weld, Persian berries and quercitron bark. The transcription of the production records used in this work may be consulted in Table S1, and the synthesis methods reproduced are described in Table 1. Interpretation of the materials used was based on our previous works [18,24,25], which allowed us to infer that the term 'whiting' used in the recipe *Yellow from Weld* corresponds to calcium carbonate (CaCO3), 'Sub. Carb. Pot', used in the same recipe, is potassium bicarbonate (KHCO3) and 'Pearlash', used in the other recipes, is potassium carbonate (K2CO3). It is important to note that the recipe *Yellow Lake. Cool tint.* refers to the use of alum (ammonia sort), however, it was chosen to use common alum (KAl(SO4)2·12H2O) in all recipes to facilitate a first comparison between them. For this reason, we also decided always to use weld flowers. The introduction of experimental variants such as using ammonium alum (NH4Al(SO4)2·12H2O) will be investigated in the future. All materials were scaled-down from industrial to laboratory scale, and quantities in British measures were converted to SI units [22,23].

pH measurements were acquired throughout the syntheses. After 1 day left to precipitate, the lakes were centrifuged for 10 min at 2400 rpm, washed with distilled water, and centrifuged again for 5 min at 3000 rpm. The lakes were air-dried and ground in an agate mortar for 15 min each.

**Table 1.** Production name, recipe and pigment code of the synthesis methods for weld lake pigments, adapted from the original text transcribed in Table S1.

§ The unique recipe code is the code from the W&N Database that identifies a database record. ¥ Although this record is a copy, its title is "Experiment with Weld for Yellow Lake for Water Colours" and is dated 6 October 1854.

#### *2.3. Paint References*

Paint references were prepared using gum arabic as a 20% solution; the pieces were ground and then added to pure water. The lake pigments were first ground in a glass mortar with pure water and then ground with the binder. The paints were applied on filter paper with a paintbrush and allowed to dry. Filter paper was selected because no additives are present such as brighteners; this was confirmed by checking the filter paper under an UV-lamp (280 nm). The paint references were analyzed by colorimetry.

#### *2.4. Equipment and Characterization Methods*


**3. Disadvantages of robotic single site surgery**

located directly adjacent to each other, without intervening fat and muscle. As a result, the umbilicus provides easy access to the abdomen, even in morbidly obese patients. Furthermore, the stalk of the umbilicus is composed primarily of fibrotic scar tissue with minimal vascularity. Consequently, most umbilical incisions are relatively bloodless [1]. In addition, single site surgery obviously eliminates the risks associated with the placement of accessory trocars, including bleeding, flank hematomas, incisional hernias, and visceral injury. The lack of additional trocars also

The most obvious advantage of single site surgery, however, is cosmesis. Even a

Robotic single site surgery offers advantages over traditional laparoscopic single site surgery. The 3-D binocular vision provided by the robotic platform allows for better depth perception and facilitates more precise surgical movements. Although the only wristed instrument is the robotic needle driver, this is also a significant advantage over all "straight stick" laparoscopic instruments. The binocular vision and wristed needle driver greatly facilitate intracorporeal suturing and knot tying. The needle driver can also be employed as a grasper and its dexterity can improve exposure for adhesiolysis or facilitate the excision of an ovarian cyst. Finally, the robotic single site platform is more ergonomic and intuitive. Intra-abdominally, the surgeon's right hand controls the right sided instrument and the left hand controls

2-3 cm incision can be hidden in the umbilicus, and it often becomes virtually invisible as it heals [4]. The poor vascularity of the umbilicus also minimizes the risk for a postoperative hematoma and virtually eliminates the risk for keloid

The most functional advantage of single site surgery is using the umbilical incision for specimen retrieval. The lack of intervening muscle and fat provides easy access to the surgical specimen. Specimen retrieval is easy, and any morcellation required is readily accomplished by bringing the specimen bag up through the

contributes to less post-operative pain [2, 3].

*Single Port Gynecologic Laparoscopic and Robotic-Assisted Surgery*

formation [5].

**Figure 1.**

**14**

*Surgeon's right hand controls the right instrument intra abdominally and vice versa.*

umbilical incision [6].

Compared to traditional multiple port robotic surgery, there are some disadvantages to the single site robotic system. The robotic single site instruments are relatively primitive. There are no advanced energy instruments such as the harmonic scalpel or bipolar transection tools built into the robotic single site system. The only unipolar tool available is the hook; the scissors do not have any unipolar power capability. In addition, the required semi-rigid flexibility of the robotic single site instruments leads to a relatively weak grasping force. This is most readily apparent when attempting to suture with the needle driver or when trying to hold tissue on tension. Furthermore, even though the needle driver is wristed, it has less range of motion than traditional robotic instruments.

Finally, the "working space" of the robotic single site system is limited compared to traditional robotic surgery. The trocar length is fixed, and the instruments cannot be retracted back any further than the tip of the trocars. This can make surgery more difficult in the setting of big pathology such as a large fibroid uterus or large ovarian cyst. In addition, in patients of short stature, the distance from the umbilicus to the pelvis is often smaller, and this can further compromise the functional workspace.

Access by the assistant surgeon can be compromised with the robotic single site system. Lateral movements can lead to repeated collisions (often coined "sword fighting") between the instruments and camera both inside the abdomen and outside. The most unencumbered instrument movements by the assistant are those performed in an anterior to posterior direction — parallel to the camera. Despite these disadvantages, the robotic single site system can readily handle most gynecologic surgery. Various techniques for overcoming these disadvantages are discussed in the "Tips and Tricks" section of this chapter.

### **4. Abdominal entry**

The initial step in any single site operation, whether robotic or laparoscopic, is the umbilical incision. Various incisions have been proposed, but the simplest, easiest, and most cosmetic approach is a midline vertical incision right through the center of the umbilicus. Local anesthesia (with or without epinephrine) is injected into the base of the umbilicus. Toothed forceps placed at the superior and inferior edges of the umbilicus are used to elevate the skin and an incision is made vertically through the center of the umbilicus. Allis clamps are then placed laterally and used to elevate the skin edges. With the edges elevated, the stalk of the umbilicus is palpated as a horizontal band of fascia in the center of the incision. Kocher clamps are then placed laterally on this fascia band, and the Allis clamps are removed. While elevating with the Kocher clamps, an incision is then made vertically in the fascia. The fascial incision is then sharply enlarged to allow the surgeon to bluntly enter the abdomen digitally. The skin and fascial incisions are then enlarged as needed. For robotic single site surgery, a 2-3 cm incision is required. This is slightly larger than what may be required for laparoscopic single site surgery, depending on the intended operation. The fascial incision should be extended vertically in both directions until it is slightly larger than the skin incision (**Figures 2**–**5**).

**Figure 4.**

*incision.*

**Figure 5.**

**17**

*Vertical fascial skin incision followed by blunt digital abdominal entry.*

*Kocher clamps placed bilaterally on the umbilical stalk which appears as a horizontal fascial band in the*

*Fundamentals of the Robotic Assisted Laparoscopic Single Port System and Utility in Minor…*

*DOI: http://dx.doi.org/10.5772/intechopen.96588*

**Figure 2.** *Vertical umbilical skin incision.*

**Figure 3.** *Allis clamps placed bilaterally on the skin edges and gently elevated.*

*Fundamentals of the Robotic Assisted Laparoscopic Single Port System and Utility in Minor… DOI: http://dx.doi.org/10.5772/intechopen.96588*

**Figure 2.**

**Figure 3.**

**16**

*Allis clamps placed bilaterally on the skin edges and gently elevated.*

*Vertical umbilical skin incision.*

*Single Port Gynecologic Laparoscopic and Robotic-Assisted Surgery*

*Kocher clamps placed bilaterally on the umbilical stalk which appears as a horizontal fascial band in the incision.*

**Figure 5.** *Vertical fascial skin incision followed by blunt digital abdominal entry.*