Search results for: eosin dye
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 62

Search results for: eosin dye

62 Fabrication of Activated Carbon from Palm Trunksfor Removal of Harmful Dyes

Authors: Eman Alzahrani

Abstract:

Date palm trees are abundant and cheap natural resources in Saudi Arabia. In this study, an activated carbon was prepared from palm trunks by chemical processes. The chemical activation was performed by impregnation of the raw materials after grinding with H3PO4 solution (63%), followed by placing of the sample solution on a muffle furnace at 400ºC for 30 min, and then at 800ºC for 10 min. The morphology of the fabricated material was checked using scanning electron microscopy that showed the rough surfaces on the carbon samples. The use of fabricated activated carbon for removal of eosin dye from aqueous solutions at different contact time, initial dye concentration, pH and adsorbent doses was investigated. The experimental results show that the adsorption process attains equilibrium within 20 min. The adsorption isotherm equilibrium was studied by means of the Langmuir and Freundlich isotherms, and it was found that the data fit the Langmuir isotherm equation with maximum monolayer adsorption capacity of 126.58 mg g-1. The results indicated that the home made activated carbon prepared from palm trunks has the ability to remove eosin dye from aqueous solution and it will be a promising adsorbent for the removal of harmful dyes from waste water.

Keywords: activated carbon, date palm trunks, H3PO4 activation, adsorption, dye removal, eosin dye, isotherm

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61 Dye Retention by a Photochemicaly Crosslinked Poly(2-Hydroxy-Ethyl-Meth-Acrylic) Network in Water

Authors: Yasmina Houda Bendahma, Tewfik Bouchaour, Meriem Merad, Ulrich Maschke

Abstract:

The purpose of this work is to study retention of dye dissolved in distilled water, by an hydrophilic acrylic polymer network. The polymer network considered is Poly (2-hydroxyethyl methacrylate) (PHEMA): it is prepared by photo-polymerization under UV irradiation in the presence of a monomer (HEMA), initiator and an agent cross-linker. PHEMA polymer network obtained can be used in the retention of dye molecules present in the wastewater. The results obtained are interesting in the study of the kinetics of swelling and de-swelling of cross linked polymer networks PHEMA in colored aqueous solutions. The dyes used for retention by the PHEMA networks are eosin Y and Malachite Green, dissolved in distilled water. Theoretical conformational study by a simplified molecular model of system cross linked PHEMA / dye (eosin Y and Malachite Green), is used to simulate the retention phenomenon (or Docking) dye molecules in cavities in nano-domains included in the PHEMA polymer network.

Keywords: dye retention, molecular modeling, photochemically crosslinked polymer network, swelling deswelling, PHEMA, HEMA

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60 Swelling Behavior of Cross-Linked Poly (2-hydroxyethyl methacrylate)

Authors: Salah Hamri, Tewfik Bouchaour, Ulrich Maschke

Abstract:

The aim of this works is the study of swelling ratio of cross-linked polymer networks poly (2-hydroxyethyl methacrylate) (PHEMA). The system composed of erythrosine and Triethanolamine, in aqueous medium, is used as photo-initiator and 1,6-Hexanediol diacrylate as cross-linker. The analysis of UV-visible and infrared spectra, which were taken at different times during polymerization/cross linking, makes it possible to obtain useful information on the reaction mechanism. The swelling behavior was study by changing the nature of solvent, dye sensitizer (erythrosine, rose Bengal and eosin), and pH of the medium. The exploitation of experimental results using Fick diffusion model is also expected and shows a good correlation between theoretical and experimental results.

Keywords: cross-linker, photo-sensitizer, polymer network, swelling ratio

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59 Gross and Histological Studies on the Thymus of the Grasscutter (Thyronomys swinderianus)

Authors: R. M. Korzerzer, J. O. Hambolu, S. O. Salami, S. B. Oladele

Abstract:

Twelve apparently healthy grasscutters between the ages of three and seven months were used for this study. The animals were purchased from local breeders in Oturkpo, Benue state, Nigeria and transported to the research laboratory in the Department of Veterinary Anatomy, Ahmadu Bello University, Zaria by means of constructed cages. The animals were divided into three groups according to their ages and acclimatised. Sacrifice was done using chloroform gaseous inhalation anaesthesia. An incision was made at the neck region and the thymus located and identified by its prominent bilateral nature. Extirpated thymuses from each animal were immediately weighed and fixed in Bouin’s fluid for 48 hours. The tissues were then prepared using standard methods. Haematoxilin and eosin was used for routine histology and Rhodamine B aniline methylene blue was for studying the architecture of the elastic and reticular fibres of the thymus. Grossly, the thymus appeared as a bilateral organ on either side of the thoracic midline. The organ size decreased consistently as the animals advanced in age. Mean ± SEM values for thymic weights were 1.23 ± 0.048 g, 0.53 ± 0.019 g and 0.30 ± 0.042 g at three, five and seven months of age respectively.

Keywords: gross, histological, thymus, grasscutter

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58 Histopathological Examination of Lung Surgery Camel in Iran

Authors: Ali Chitgar

Abstract:

Respiratory infections including diseases in camels are important not only because of the threat of animal health but also to reduce their production. Since that deal with respiratory problems and their treatment requires adequate knowledge of the existing respiratory problems, unfortunately, there is limited information about the species of camels. This study aimed to identify lung lesions camels slaughtered in a slaughterhouse more important was performed using histopathology. Respiratory camels (n = 477) was examined after the killing fully and tissue samples were placed in 10% formalin. The samples and histological sections using hematoxylin and eosin staining and color were evaluated. In this study 79.6 % (236 of 477 samples) of the samples was at least a lung lesion. Rate acute interstitial pneumonia, chronic interstitial pneumonia, bronchopneumonia, bronchiolitis, an inflammation of the pleura and 52.8 % respectively atelectasis (236 of 477 samples), 5.4 % (24 of 477 samples), 7.8 % (35 of 477 samples), 6.7 % (30 of 477 samples), 3.4 % (15 of 477 samples) and 15.2% (68 of 477 samples). The lung lesions, acute interstitial pneumonia and bronchopneumonia in autumn winter rather than spring and summer (p <0/05) and as a result, this study showed that high rates of lung lesions in the camel population. Waste higher results in cold seasons (fall and winter) shows.

Keywords: camel, surgery, histopathology, breathing organ

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57 Genotoxic Effect of Tricyclieandidepressant Drug “Clomipramine Hydrochloride’ on Somatic and Germ Cells of Male Mice

Authors: Samia A. El-Fiky, F. A. Abou-Zaid, Ibrahim M. Farag, Naira M. Efiky

Abstract:

Clomipramine hydrochloride is one of the most used tricyclic antidepressant drug in Egypt. This drug contains in its chemical structure on two benzene rings. Benzene is considered to be toxic and clastogenic agent. So, the present study was designed to assess the genotoxic effect of Clomipramine hydrochloride on somatic and germ cells in mice. Three dose levels 0.195 (Low), 0.26 (Medium), and 0.65 (High) mg/kg.b.wt. were used. Seven groups of male mice were utilized in this work. The first group was employed as a control. In the remaining six groups, each of the above doses was orally administrated for two groups, one of them was treated for 5 days and the other group was given the same dose for 30 days. At the end of experiments, the animals were sacrificed for cytogenetic and sperm examination as well as histopathological investigations by using hematoxylin and eosin stains (H and E stains) and electron microscope. Concerning the sperm studies, these studies were confined to 5 days treatment with different dose levels. Moreover, the ultrastructural investigation by electron microscope was restricted to 30 days treatment with drug doses. The results of the dose dependent effect of Clomipramine showed that the treatment with three different doses induced increases of frequencies of chromosome aberrations in bone marrow and spermatocyte cells as compared to control. In addition, mitotic and meiotic activities of somatic and germ cells were declined. The treatments with medium or high doses were more effective for inducing significant increases of chromosome aberrations and significant decreases of cell divisions than treatment with low dose. The effect of high dose was more pronounced for causing such genetic deleterious in respect to effect of medium dose. Moreover, the results of the time dependent effect of Clomipramine observed that the treatment with different dose levels for 30 days led to significant increases of genetic aberrations than treatment for 5 days. Sperm examinations revealed that the treatment with Clomipramine at different dose levels caused significant increase of sperm shape abnormalities and significant decrease in sperm count as compared to control. The adverse effects on sperm shape and count were more obviousness by using the treatments with medium or high doses than those found in treatment with low dose. The group of mice treated with high dose had the highest rate of sperm shape abnormalities and the lowest proportion of sperm count as compared to mice received medium dose. In histopathological investigation, hematoxylin and eosin stains showed that, the using of low dose of Clomipramine for 5 or 30 days caused a little pathological changes in liver tissue. However, using medium and high doses for 5 or 30 days induced severe damages than that observed in mice treated with low dose. The treatment with high dose for 30 days gave the worst results of pathological changes in hepatic cells. Moreover, ultrastructure examination revealed, the mice treated with low dose of Clomipramine had little differences in liver histological architecture as compared to control group. These differences were confined to cytoplasmic inclusions. Whereas, prominent pathological changes in nuclei as well as dilated of rough Endoplasmic Reticulum (rER) were observed in mice treated with medium or high doses of Clomipramine drug. In conclusion, the present study adds evidence that treatments with medium or high doses of Clomipramine have genotoxic effects on somatic and germ cells of mice, as unwanted side effects. However, the using of low dose (especially for short time, 5 days) can be utilized as a therapeutic dose, where it caused relatively similar proportions of genetic, sperm, and histopathological changes as those found in normal control.

Keywords: clomipramine, mice, chromosome aberrations, sperm abnormalities, histopathology

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56 Genotoxic Effect of Tricyclic Antidepressant Drug “Clomipramine Hydrochloride’ on Somatic and Germ Cells of Male Mice

Authors: Samia A. El-Fiky, Fouad A. Abou-Zaid, Ibrahim M. Farag, Naira M. El-Fiky

Abstract:

Clomipramine hydrochloride is one of the most used tricyclic antidepressant drug in Egypt. This drug contains in its chemical structure on two benzene rings. Benzene is considered to be toxic and clastogenic agent. So, the present study was designed to assess the genotoxic effect of Clomipramine hydrochloride on somatic and germ cells in mice. Three dose levels 0.195 (Low), 0.26 (Medium), and 0.65 (High) mg/kg.b.wt. were used. Seven groups of male mice were utilized in this work. The first group was employed as a control. In the remaining six groups, each of the above doses was orally administrated for two groups, one of them was treated for 5 days and the other group was given the same dose for 30 days. At the end of experiments, the animals were sacrificed for cytogenetic and sperm examination as well as histopathological investigations by using hematoxylin and eosin stains (H and E stains) and electron microscope. Concerning the sperm studies, these studies were confined to 5 days treatment with different dose levels. Moreover, the ultrastructural investigation by electron microscope was restricted to 30 days treatment with drug doses. The results of the dose dependent effect of Clomipramine showed that the treatment with three different doses induced increases of frequencies of chromosome aberrations in bone marrow and spermatocyte cells as compared to control. In addition, mitotic and meiotic activities of somatic and germ cells were declined. The treatments with medium or high doses were more effective for inducing significant increases of chromosome aberrations and significant decreases of cell divisions than treatment with low dose. The effect of high dose was more pronounced for causing such genetic deleterious in respect to effect of medium dose. Moreover, the results of the time dependent effect of Clomipramine observed that the treatment with different dose levels for 30 days led to significant increases of genetic aberrations than treatment for 5 days. Sperm examinations revealed that the treatment with Clomipramine at different dose levels caused significant increase of sperm shape abnormalities and significant decrease in sperm count as compared to control. The adverse effects on sperm shape and count were more obviousness by using the treatments with medium or high doses than those found in treatment with low dose. The group of mice treated with high dose had the highest rate of sperm shape abnormalities and the lowest proportion of sperm count as compared to mice received medium dose. In histopathological investigation, hematoxylin and eosin stains showed that, the using of low dose of Clomipramine for 5 or 30 days caused a little pathological changes in liver tissue. However, using medium and high doses for 5 or 30 days induced severe damages than that observed in mice treated with low dose. The treatment with high dose for 30 days gave the worst results of pathological changes in hepatic cells. Moreover, ultrastructure examination revealed, the mice treated with low dose of Clomipramine had little differences in liver histological architecture as compared to control group. These differences were confined to cytoplasmic inclusions. Whereas, prominent pathological changes in nuclei as well as dilated of rough Endoplasmic Reticulum (rER) were observed in mice treated with medium or high doses of Clomipramine drug. In conclusion, the present study adds evidence that treatments with medium or high doses of Clomipramine have genotoxic effects on somatic and germ cells of mice, as unwanted side effects. However, the using of low dose (especially for short time, 5 days) can be utilized as a therapeutic dose, where it caused relatively similar proportions of genetic, sperm, and histopathological changes as those found in normal control.

Keywords: chromosome aberrations, clomipramine, mice, histopathology, sperm abnormalities

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55 Acute Toxic Effects of Zn(SO4) on Gill and Liver Tissues of Fresh Water Catfish Clarias batrachus (L.)

Authors: Muneesh Kumar, Rajesh Kumar, Sangeeta Devi

Abstract:

Heavy metals are a major problem because they are toxic and tend to accumulate in living organisms. This study was carried out with the aims of studying on histopathology of Zn(SO4) toxicity on gill and liver tissues of catfish (Clarias batrachus) within the period of 96 h. Totally, 140 fishes with mean weight 50±10 g were stocked in 12 aquariums with capacity of 200 L water and divided in to 3 trails including control, 4 ppm and 8 ppm of Zn with 3 replicates. Tissue samples were fixed by bouin’s solution and sectioned in 7 μm based on histological regular method and stained with Hematoxylin and Eosin (H&E) method for microscopic study within the period of 96 h. Results showed some damaged such as hyperplasia, telangiectasis and edema, necrosis of second filaments, jerky movement, aneurism, hyperemia and fusion of second filaments in gills; and cell atrophy, necrosis, fatty degeneration, hyperemia and bile stagnation at different treatments in comparison with control. Gill and liver tissue damages were severed with the increase of Zn concentration and days. Therefore, Zn had acute toxicity effects on gill and liver tissues in Catfish at 5 and 10 ppm concentrations.

Keywords: gill, liver, histopathology, zinc, Clarias batrachus

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54 The Change in the Temporomandibular Joint Bone in Osteoarthritis Induced Mice

Authors: Boonyalitpun P., Pruckpattranon P., Thonghom A., Rotpenpian N.

Abstract:

Osteoarthritis is a musculoskeletal and neuromuscular abnormality, masticatory muscle, and other tissue that causes pain and breaks down the articular surface of the temporomandibular joint (TMJ). The aim of this study is to investigate the change in the mandibular condyle, in terms of thickness and porosity, and osteoclast marker in the mandibular condyle of TMJ induced osteoarthritis mice (TMJ-OA mice). We investigated the bony changes in the TMJ structure of a complete Freund adjuvant (CFA)-injected TMJ in a mice model over 28 days. On day 28, we observed any change in the TMJ by a micro computed tomography scan (micro-CT scan) in the parameters of trabecular microarchitecture. Then we studied the thickness of the condyles by hematoxylin and eosin staining. Moreover, we calculated the area around the TMJ’s condylar head containing the osteoclast expression by TRAP (Tartrate-resistant acid phosphatase) immunohistochemistry staining. The result found that the parameter of a micro-CT scan was no different from microarchitecture in the TMJ compared with the control group; however, mandibular condyles of the TMJ-OA group was significantly thinner than the control groups, and the osteoclast expression significantly increased in the TMJ-OA group. Therefore, our findings suggest that CFA-induced TMJ-OA represents an expression of osteoclast mandibular condyle of the TMJ, which is the proposed mechanism for a TMJ-OA model.

Keywords: condyle, osteoarthritis, osteoclast, temporomandibular joint

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53 Physicochemical Attributes of Pectin Hydrogel and Its Wound Healing Activity

Authors: Nor Khaizan Anuar, Nur Karimah Aziz, Tin Wui Wong, Ahmad Sazali Hamzah, Wan Rozita Wan Engah

Abstract:

The physicochemical attributes and wound healing activity of pectin hydrogel in rat models, following partial thickness thermal injury were investigated. The pectin hydrogel was prepared by solvent evaporation method with the aid of glutaraldehyde as crosslinking agent and glycerol as plasticizer. The physicochemical properties were mainly evaluated using differential scanning calorimetry (DSC) and Fourier transform infrared (FTIR) spectroscopy, while the wound healing activity was examined by the macroscopic images, wound size reduction and histological evaluation using haematoxylin and eosin (H&E) stain for 14 days. The DSC and FTIR analysis suggested that pectin hydrogel exhibited higher extent of polymer-polymer interaction at O-H functional group in comparison to the unprocessed pectin. This was indicated by the increase of endothermic enthalpy values from 139.35 ± 13.06 J/g of unprocessed pectin to 156.23 ± 2.86 J/g of pectin hydrogel, as well as the decrease of FTIR wavenumber corresponding to O-H at 3432.07 ± 0.49 cm-1 of unprocessed pectin to 3412.62 ± 13.06 cm-1 of pectin hydrogel. Rats treated with pectin hydrogel had significantly smaller wound size (Student’s t-test, p<0.05) when compared to the untreated group starting from day 7 until day 14. H&E staining indicated that wounds received pectin hydrogel had more fibroblasts, blood vessels and collagen bundles on day 14 in comparison to the untreated rats.

Keywords: pectin, physicochemical, rats, wound

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52 Comparison of the Effects of Fresh Leaf, Septum and Peel Extracts of Walnut on Blood Glucose and Pancreatic Structure

Authors: Tahmineh Hasanzadeh, Afshin Farahbakhsh

Abstract:

There is some report about the hypoglycemic effect of Juglans rejia L. leaf in alloxan induced diabetic rats and hypoglycemic effect of its fruit peel administered intraperitoneally.In Iranian traditional medicine, septum of walnut shell (SWS) was recommended to reduce blood glucose. For this purpose, 41 male bulb/C mice 25-30 gm were divided into five groups. All the animals received IP injection of streptozotocin (STZ) (220 mg/kg). Two weeks later, the diabetic animals were received daily oral treatment of normal saline and aqueous extract of SWS (200, 400, 600 and 800 mg/kg) respectively for four weeks. Blood samples were taken from retro orbital sinus before the start of the experiment and repeated each two weeks. At the end of the experiment, the animals were sacrificed and the pancreatic tissues were fixed, prepared and stained by Hematoxylin-Eosin for light microscope studies. The results showed that in each group, the SWS extract reduced blood glucose in a long time (p < 0.05). metabolic extract in STZ- induced diabetic rats, which was accompanied by the hypoglycemic effect of leaf extract. However, this effect should be determined with scientific researches. Therefore, the aim of this study is to evaluate the effect of the aqueous extract of SWS on blood glucose and histopathological structure of pancreas.

Keywords: septum of walnut, blood glucose, pancreas, diabetes, walnut leaf, walnut peel, insulin

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51 In vitro Effects of Amygdalin on the Functional Competence of Rabbit Spermatozoa

Authors: Marek Halenár, Eva Tvrdá, Tomáš Slanina, Ľubomír Ondruška, Eduard Kolesár, Peter Massányi, Adriana Kolesárová

Abstract:

The present in vitro study was designed to reveal whether amygdalin (AMG) is able to cause changes to the motility, viability and mitochondrial activity of rabbit spermatozoa. New Zealand White rabbits (n = 10) aged four months were used in the study. Semen samples were collected from each animal and used for the in vitro incubation. The samples were divided into five equal parts and diluted with saline supplemented with 0, 0.5, 1, 2.5 and 5 mg/mL AMG. At times 0h, 3h and 5h spermatozoa motion parameters were assessed using the SpermVision™ computer-aided sperm analysis (CASA) system, cell viability was examined with the metabolic activity (MTT) assay, and the eosin-nigrosin staining technique was used to evaluate the viability of rabbit spermatozoa. All AMG concentrations exhibited stimulating effects on the spermatozoa activity, as shown by a significant preservation of the motility (P<0.05 with respect to 0.5 mg/mL and 1 mg/mL AMG; Time 5 h) and mitochondrial activity (P< 0.05 in case of 0.5 mg/mL AMG; P< 0.01 in case of 1 mg/mL AMG; P < 0.001 with respect to 2.5 mg/mL and 5 mg/mL AMG; Time 5 h). None of the AMG doses supplemented had any significant impact of the spermatozoa viability. In conclusion, the data revealed that short-term co-incubation of spermatozoa with AMG may result in a higher preservation of the sperm structural integrity and functional activity.

Keywords: amygdalin, CASA, mitochondrial activity, motility, rabbits, spermatozoa, viability

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50 Carvacrol Attenuates Lung Injury in Rats with Severe Acute Pancreatitis

Authors: Salim Cerig, Fatime Geyikoglu, Pınar Akpulat, Suat Colak, Hasan Turkez, Murat Bakir, Mirkhalil Hosseinigouzdagani, Kubra Koc

Abstract:

This study was designed to evaluate whether carvacrol (CAR) could provide protection against lung injury by acute pancreatitis development. The rats were randomized into groups to receive (I) no therapy; (II) 50 μg/kg cerulein at 1h intervals by four intraperitoneal injections (i.p.); (III) 50, 100 and 200 mg/kg CAR by one i.p.; and (IV) cerulein+CAR after 2h of cerulein injection. 12h later, serum samples were obtained to assess pancreatic function the lipase and amylase values. The animals were euthanized and lung samples were excised. The specimens were stained with hematoxylin-eosin (H&E), periodic acid–Schif (PAS), Mallory's trichrome and amyloid. Additionally, oxidative DNA damage was determined by measuring as increases in 8-hydroxy-deoxyguanosine (8-OH-dG) adducts. The results showed that the serum activity of lipase and amylase in AP rats were significantly reduced after the therapy (p<0.05). We also found that the 100 mg/kg dose of CAR significantly decreased 8-OH-dG levels. Moreover, the severe pathological findings in the lung such as necrosis, inflammation, congestion, fibrosis, and thickened alveolar septum were attenuated in the AP+CAR groups when compared with AP group. Finally, the magnitude of the protective effect on lung is certain, and CAR is an effective therapy for lung injury caused by AP.

Keywords: antioxidant activity, acute pancreatitis, carvacrol, experimental, lung injury, oxidative DNA damage

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49 Antigastritic Effect of Starch from Manihot utilissima on Male Wistar Rats Induced Aspirin

Authors: Naela Nabiela, Ahmad Hilmi Fahmi, M. Sukron, Ayu Elita Sari, Yusran, Suparmi

Abstract:

Aspirin is one of NSAIDs (non-steroid inflammatory drugs), can cause gastric ulcer as an side effect of prolonged consumption. The effort to prevent the increase of gastric HCl level can by treating with amylopectin was reported that can cover the gastric mucose. However, the effect of amylopectin in starch from Manihot utilissima which is believed as traditional treatment gastric ulcer have not been clear yet. This study was conducted to determine the effect of starch formed as syrup to HCl level and gastric histopatology. This experiment post test only control group design used 42 male wistar rats divided into 7 groups. All groups, except first group, were induced by 60 mg/100gBW/day aspirin for 3 days. The following day for 2 days each group was treated by starch syrup at dosed 0.45% w/v, 0.9% w/v, 1.8% w/v, 0% w/v, and sucralfate. Respectively, HCl level were measured by acidi-alkalimetri titration method, while the gastric histopathology were prepared by hematoxylin-eosin staining. The result shows that aspirin induction can increase the HCl level as 0,00767 N. Starch syrup at dose 1.8% w/v was effective to reduce HCl level and the grade of second gastric necrosis. It can be conclude that starch syrup is potention as a treatment to cure gastric ulcer caused by NSAIDs side effect.

Keywords: concentration of HCl stomach, gastric histopathology, gastritis, starch

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48 The Effects of Red Onion Extract (Allium ascalonicum L.) in the Pulmonary Histopathological Lesions of Layer Chickens at 47 Days Old Raised in the Battery Cage

Authors: R. N. Nataria, A. D. Paryuni, R. Wasito

Abstract:

Layer farms in Indonesia have still obstacles to increasing their productivity, especially due to poultry diseases. The red onion (Allium ascalonicum L.) is a plant that contains flavonoid and saponin. Flavonoid is useful as anti-inflammatory and antioxidant while saponin is useful as antivirus, anti-inflammatory, antifungal, and immunomodulator. This study aimed to know and determine the effect of onion extracts to pulmonary histopathological lesions in layer chickens which raised in the battery cage. This study used eighteen layer chickens at seventeen days old. The eighteen layer chickens were divided into three groups of six each, namely without administration of red onion extract (Group I), with administration red onion extract through drinking water (Group II) and with administration red onion extract peroral (Group III). Every ten days, six chickens were necropsied and then the lungs were processed for histopathological preparations and stained with routine hematoxylin and eosin. The results showed that the lungs of the Group I had severe congestion and diffuse hemorrhages. In Group II, lungs had moderate congestion and hemorrhages. In group III, lungs had mild congestion and hemorrhages. It is concluded, that red onion extract apparently has reduced the lungs lesions in layer chickens.

Keywords: histopathological lesions, layers, lungs, poultry diseases, red onion extract

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47 Histochemical Localization of Hepatitis B Surface Antigen in Hepatocellular Carcinoma: An Evaluation of Two Staining Techniques in a Tertiary Hospital in Calabar, Nigeria

Authors: Imeobong Joseph Inyang, Aniekan-Augusta Okon Eyo, Abel William Essien

Abstract:

Hepatitis B virus (HBV) is one of the known human carcinogens. The presence of HBsAg in liver tissues indicates active viral replication. More than 85% of Hepatocellular Carcinoma (HCC) cases occur in countries with increased rates of chronic HBV infection. An evaluation study to determine the relationship between positivity for HBsAg and development of HCC and its distribution between age and gender of subjects was done. Shikata Orcein and Haematoxylin and Eosin (H&E) staining techniques were performed on liver sections. A total of 50 liver tissue specimens comprising 38 biopsy and 12 post-mortem specimens were processed. Thirty-five of the 50 specimens were positive for HBsAg with Orcein stain whereas only 16 were positive with H&E stain, and these were also positive with Orcein stain, giving an HBsAg prevalence of 70.0% (35/50). The prevalence of HCC in the study was 56.0% (28/50), of which 21 (75.0%) cases were positive for HBsAg, 18 (64.3%) were males while 10 (35.7%) were females distributed within the age range of 20-70 years. The highest number of HBsAg positive HCC cases, 7/21 (33.3%) occurred in the age group 40-49 years. There was no relationship in the pattern of distribution of HCC between age and gender using the Pearson correlation coefficient (r = 0.0474; P < 0.05). HBV infection predisposed to HCC. Orcein technique was more specific and is therefore recommended for screening of liver tissues where facilities for immunohistochemistry are inaccessible.

Keywords: Hepatitis B. surface antigen, hepatocellular carcinoma, orcein, pathology

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46 Modifiable Poly Methacrylic Acid-Co-Acrylonitrile Microgels Fabricated with Cu and Co Nanoparticles for Simultaneous Catalytic Reduction of Multiple Compounds

Authors: Muhammad Ajmal, Muhammad Siddiq, Nurettin Sahiner

Abstract:

We prepared poly(methacrylic acid-co-acrylonitrile) (p(MAc-co-AN)) microgels by inverse suspension polymerization, and converted the nitrile groups into amidoxime groups to obtain more hydrophilic amidoximated poly(methacrylic acid-co-acrylonitile) (amid-p(MAc-co-AN)) microgels. Amid-microgels were used as microreactors for in situ synthesis of copper and cobalt nanoparticles. Cu (II) and Co (II) ions were loaded into microgels from their aqueous metal salt solutions and then converted to corresponding metal nanoparticle (MNP) by treating the loaded metal ions with sodium borohydride (NaBH4). The characterization of the prepared microgels and microgel metal nanoparticle composites was carried out by SEM, TEM and TG analysis. The amounts of metal nanoparticles within microgels were estimated by AAS measurements by dissolving the MNP entrapped within microgels by concentrated HCl acid treatment. Catalytic performances of the prepared amid-p(MAc-co-AN)-M (M: Cu, Co) microgel composites were investigated by using them as catalyst for the degradation of cationic and anionic organic dyes such as eosin Y (EY), methylene blue (MB) and methyl Orange (MO), and for the reduction of nitro aromatic pollutants like 2-nitrophenol (2-NP) and 4-nitrophenol (4-NP) to their corresponding amino phenols. Here, we also report for the first time, the simultaneous degradation/reduction of MB, EY, and 4-NP by amid-p(MAc-co-AN)-Cu microgel composites. Different parameters affecting the reduction rates such as metal types, amount of catalysts, temperature and the amount of reducing agent were investigated.

Keywords: microgels, nanoparticles, catalyst, pollutants

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45 Cimifugin Inhibited Th2-Type Allergic Contact Dermatitis

Authors: Xiaoyan Jiang, Huizhu Wang, Lili Gui, Dandan Shen, Xiao Wei, Xi Yu, Hailiang Liu, Min Hong

Abstract:

Objective: Applicate FITC to establish Th2-type allergic contact dermatitis model, and study the effect and mechanism of Cimifugin on Th2-type allergic contact dermatitis. Methods: The Balb/c mice were sensitized with painting 80 ul of 1.5% FITC onto the shaved abdomen skin at DAY1 and DAY2. The animals were challenged on their right ears with 20 ul of 0.6% FITC, and the left ears were painted with solvent alone at day 6, mice were administered cimifugin for 7 days. 24h later, ear swelling was noted, and the infiltration of eosinophils was investigated by hematoxylin and eosin (H&E) staining. while part of the ear tissue homogenates prepared for detecting interleukin-4 levels by ELISA .Mice were administered cimifugin In the initial stage of the above model for 5 days(-1DAY—DAY3), ear tissue were homogenized to detect IL-33 levels by ELISA. Results: Cimifugin 25mg/kg, 50mg/kg inhibited mouse ear swelling, ear histopathology showed that mice given Cimifugin has significantly reduced levels of local tissue fluid exudation, congestion, infiltration of lymphocytes, and other inflammatory conditions compared with the model group. At the same time, it has significantly reduce of Th2 cytokines IL-4 in the mouse ear tissue homogenate. Data of the initial stage shows that 12.5mg/kg, 50mg/kg Cimifugin significantly inhibited IL-33 levels. Conclusion: Cimifugin inhibit FITC-induced Th2-type allergic contact dermatitis, and its mechanism may be related to inhibition of IL-33.

Keywords: cimifugin, allergic contact dermatitis, Th1/Th2, IL-33

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44 Antioxidant Juice Prevents UV- Induced Skin Damage in Rats

Authors: S. P. Gomes, D. C. Goncalves, E. Ribeiro, M. C. L. Seelaender

Abstract:

Skin is susceptible to photo damage induced by exposure to sunlight, or ultraviolet (UV) radiation, which induces breakdown of extracellular matrix, DNA degradation, skin cell lesion and apoptosis, and development of cancer. Phytonutrients demonstrate protective effects against UV damage. The purpose of this study was evaluating the effect of an antioxidant juice (AJ) contaning Brazilian natural products upon skin damage. The juice was produced by Metabolics®. Male Wistar rats were divided in 4 groups: Animals receiving the antioxidant juice (AJ): orange, carrot, honey, tomato extract, avocado, ginger and camu-camu (Brazilian fruit, a major source of vitamin C) ad libitum for 21 days; or water (C), subdivided in groups exposed or not to UV radiation for 2 non consecutive days, during five hours each day, after 15 days of juice supplementation. On the 22nd day, rats were killed by decapitation and epithelium samples from the dorsal skin removed, fixed in bouin and embedded in paraffin. The sections were stained with hematoxylin and eosin or mallory and picrosirius red. Isolated DNA was submitted to electrophoresis (1.8% agarose gel, 0.5% ethidium bromide). UV radiation significantly induced sunburn of superficial epithelial cells of C, AJ treatment reduced this effect. Collagen changes were observed in UV groups, yet AJ treatment prevented collagen degradation. UV radiation induced significant DNA degradation, in C, which was prevented by AJ treatment. The antioxidant juice consumed chronically protected against acute skin damage.

Keywords: nutraceuticals, antioxidants, photoprotection, uv radiation

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43 Anatomical Studies on the Spleen and Mesenteric Lymph Node of the Grasscutter

Authors: R. M. Korzerzer, J. O. Hambolu, S. O. Salami, S. B. Oladele

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The grasscutter (Thryonomys swinderianus) has become an important source of protein and income to rural dwellers in most West African countries including Nigeria. Twelve apparently healthy grasscutters consisting of six males and six females between the ages of three and seven months were obtained from rural dwellers in Benue state and used for this study. The animals were transported by means of constructed cages to the Department of Veterinary Anatomy, Ahmadu Bello University, Zaria and sacrificed using chloroform inhalation gaseous anaesthesia by suffocation. The spleen and mesenteric lymph nodes were extirpated and the tissues prepared using standard methods, haematoxilin and eosin stain was used for routine histology, while Rhodamine B-aniline-methylene blue stain was used for staining reticular and elastic fibres. The spleen was dark red in colour and roughly triangular in outline, and was observed to increase consistently with age, maximum values were recorded at seven months of age in both males and females. Mean ± SEM values for splenic weights were 0.67 ± 0.09 g, 1.65 ± 0.35 g and 2.31 ± 0.06 g at three, five and seven months of age, respectively. The percentage ratio of splenic weight to body weight was 0.1%. Histologically, the germinal centres revealed three zones; the germinal centre, cortical layer and the marginal zone. The mesenteric lymph nodes were constantly bean shaped and appeared as opaque white masses which resemble fat but were distinguished from fat by their pearly glossy nature. The mean ± SEM values for mesenteric lymph node weights were 0.056 ± 0.005 g, 0.143 ± 0.034 g and 0.1600 ± 0.023 g at three, five and seven months of age, respectively.

Keywords: anatomical, spleen, mesenteric lymph node, grasscutter

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42 Pathological Observations of Intestinal Coccidiosis in Camel (Camelus dromedarius)

Authors: Abhilasha Dadhich, Manisha Mathur, Sanjay Kumar, Hemant Dadhich

Abstract:

The camel (Camelus dromedarius) is an important animal component of the fragile desert eco-system of India. Apart from others, impaired milk and meat production decrease in performance and even death are some of the major consequences of parasitic disease like coccidiosis in camel. Coccidiosis which is an acute invasion and destruction of intestinal mucosa by protozoa of the genera Eimeria or isospora spp. Post-Mortem examinations of 5 carcasses of dromedary of different age groups aged from 2 to 5 years were conducted. The history indicated that the camels were suffering from diarrhoea, dysentery, pyrexia, inappetence, weight loss, and emaciation. Post mortem examinations showed macroscopic and microscopic alterations in the small intestine, particularly in jejunum and ileum regions. The mucosae were congested, and haemorrhagic on which there were numerous whitish-grey nodular foci were observed. The affected intestinal tissue specimens were preserved in 10% formal saline and processed mechanically for paraffin embedding by acetone and benzene technique. The sections were stained with haematoxylin and eosin method of staining for histopathological examinations. Histologically, typical lesions such as congestion and haemorrhages were present. The intestinal villi were oedematous; mucosa degenerated and desquamated, along with infiltration of eosinophils and macrophages. Crypts of lieberkuhn were obliterated due to presence of schizonts in lamina propria. Older camels served as the source of spread of coccidial infection and were also predisposed to secondary infections.

Keywords: camel, coccidiosis, Eimeria, histopathology

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41 Detection of Leptospira interrogans in Kidney and Urine of water Buffalo and its Relationship with Histopathological and Serological Findings

Authors: M. R. Haji Hajikolaei, A. A. Nikvand, A. R. Ghadrdan, M. Ghorbanpoor, B. Mohammadian

Abstract:

This study was carried out on water buffalo for detection of Leptospira interrogans in kidney and urine and its relationship with serological findings. Blood, urine and kidney samples were taken immediately after slaughter from 353 water buffalos at Ahvaz abattoir in Khouzestan province, Iran. Sera were initially screened at serum dilution of 1:100 against seven live antigens of Leptospira interrogans: pomona, hardjo, ballum, icterohemorrhagiae, tarasovi, australis and grippotyphosa using the microscopic agglutination test (MAT) and sera with positive results were titrated against reacting antigens in serial twofold dilution from 1:100 to 1:800. The samples of kidney were embedded in paraffin wax and 5µm thick sections were stained routinely with Haematoxylin and Eosin (H&E). Polymerase chain reaction (PCR) examination was done on urine and kidney by using LipL32 gene primers. Antibodies against one or more serovars at dilution >:100 were detected in sera. The most frequent reactor was hardjo (56.2%), followed by pomona (52.3%), australis (9.8%), tarassovi (5.9%), grippotyphosa (4.5%) and icterohaemorrhagiae (3.9%). The L. interrogans were detected in 43 (12.2%) of examined buffaloes, so that 26 (8.2%) of kidney tissues, 14 (4.8%) of urine samples separately and 3 (0.84%) of both kidney and urine samples were positive in PCR. From 153 (43.3%) buffaloes with positive MAT, 24 cases were positive by PCR of kidney and/or urine samples, synchronously. Renal lesions such as interstitial nephritis, acute tubular necrosis (ATN), pyelonephritis, glomerolonephritis, renal fibrosis and hydronephrosis were found in 128 (36.3%) cases. Statistical analysis indicated that there was no significant association between results of MAT, PCR and interstitial nephritis.

Keywords: leptospiral infection, PCR, MAT, histopathology, river buffalo

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40 An Electron Microscopic Study of Developing Human Fetal Pancreas

Authors: Gupta Renu, T. S. Roy

Abstract:

Introduction: For the prospect of successful replacement therapies in treatment of Diabetes mallitus it is necessary to know events occurring during normal human pancreas development. Literature of human pancreas development are few in number as well as mainly related to first trimester because of ethical and technical difficulties. So the study was conducted on 12 fetuses from 12 gestational weeks (GW) to 5 months of infant to know normal development of exocrine and endocrine part of human pancreas. Material and Methods: Human fetalpancreases were screened by haematoxyline and eosin staining and done electron microscopy for suitable specimens to know ultrastructural detail of fetal pancreas. Results:It was observed arborized tubules, the cells budding out from these tubules differentiated into primitive acini and islets in 12thGW. At 14 weeks scanty granules were observed in the endocrine cells which coincided with the capillary invasion of the islets. The ducts and acini were surrounded by well-organized connective tissue. The acinihad elongated cells, small amount of cytoplasm and large open face euchromatic nuclei with single nucleolus. The mature form of islets of Langerhans was observed close to the acini and duct in 20 GW fetus. Connective tissue around the duct was well organized.No significant developmental change was observed early postnatal, infant. Conclusion: The development of both component exocrine as well as endocrine part of human fetal pancreas was studied by light and electron microscopy. Observations suggested that the fetal pancreas contained mainly ducts, few acini, many centroacinar cells, and large undifferentiated tissue.

Keywords: gestational weeks (GW), acini, islets of Langerhans, ducts

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39 Rebamipide Retards CCL4 Induced Hepatic Fibrosis: A Role of PGE2

Authors: Alaa E. El-sisi, Sherin Zakaria

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Rebamipide is an antiulcer drug with unique properties such as anti-inflammatory action. It induces endogenous prostaglandin e2 (PGE2). PGE2 is considered as a potent physiological suppressor of liver fibrosis. Aim of study: This study investigated the effect of rebamipide on hepatic fibrosis. Material and Method: Hepatic fibrosis was induced by intraperitoneal injections (IP) injection of CCl4 (0.45 mL/kg) in corn oil 1:5 twice a week for 4 weeks. Rats were divided into four groups as follow: Group 1 treated with CCL4 only, group 2 and 3 treated with CCL4 and rebamipide 60 mg/kg/day (group2) or 100 mg/kg/day (group3), and the fourth group was considered as control group and treated with vehicles. ALT, AST, and Bilirubin were assayed in serum. Antioxidant markers such as malondialdhyde (MDA) and superoxide dismutase (SOD) and fibrotic markers such as hyaluronic acid (HA) and procollagen-III (procol-III) were evaluated in liver tissues. IL-10 as well as PGE2 were also assayed in liver tissues. Pathologic changes in the liver were detected by hematoxylin and eosin staining. Collagen precipitation in liver tissues was visualized using masson trichrom stain. Results: Rebamipide inhibit CCL4 induced increase in ALT and AST significantly (p < 0.05). Rebamipide exerted an antioxidant effect as it inhibits CCL4 induced increased MDA level and decreased SOD activity. Fibrotic markers assay revealed that repamipide (60 or 100 mg/kg/day) decreased the level of procol-III and HA compared to CCl4 (p < 0.05). Oral administration of Rebamipide was associated with a significant increase (p < 0.05) of PGE2 and IL-10. Rebamipide especially at the dose of (100 mg/kg/day) restores liver histology structure and abolish collagen precipitation in liver tissues. Conclusion: Rebamipide retards hepatic fibrosis induced by CCL4 may be through the induction of PGE2 level.

Keywords: fibrotic markers, hepatic fibrosis, PGE2, rebamipide

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38 Histopathological Examination of BALB/C Mice Receiving Strains of Acinetobacter baumannii Resistant to Colistin Antibiotic

Authors: Shahriar Sepahvand, Mohammad Ali Davarpanah

Abstract:

Infections caused by Acinetobacter baumannii are among the common hospital-acquired infections that have seen an increase in antibiotic resistance in recent years. Colistin is the last treatment option against this pathogen. The aim of this study is to investigate the histopathology of BALB/C mice receiving sensitive and resistant strains of Acinetobacter baumannii to colistin. A total of 68 female laboratory mice weighing 30 to 40 grams of the BALB/C breed were studied in this research for three weeks under appropriate laboratory conditions in terms of food and environment. The experimental groups included: control group, second group, third group, fourth group. Lung, liver, spleen, and kidney tissues were removed from anesthetized mice and, after washing in physiological serum, were fixed in 10% formalin for 14 days. For dehydration, alcohol with ascending degrees of 70, 80, 90, and 100 was used. After clearing and soaking in paraffin, the samples were embedded in paraffin. Then, sections with a thickness of 5 microns were prepared and, after staining by hematoxylin-eosin, the samples were ready for study with a light microscope. In liver, spleen, lung, and kidney tissues of mice receiving the colistin-sensitive strain of Acinetobacter baumannii, infiltration of inflammatory cells and hyperemia were observed compared to control group mice. Liver and lung tissues of mice receiving strains of Acinetobacter baumannii resistant to colistin showed tissue destruction in addition to infiltration of inflammatory cells and hyperemia, with more destruction observed in lung tissue.

Keywords: acinetobacter baumannii, colistin antibiotic, histopathological examination, resistant

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37 Protective Effect of Saponin Extract from the Root of Garcinia kola (Bitter Kola) against Paracetamol-Induced Hepatotoxicity in Albino Rats

Authors: Alli Smith Yemisi Rufina, Adanlawo Isaac Gbadura

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Liver disorders are one of the major problems of the world. Despite its frequent occurrence, high morbidity, and high mortality, its medical management is currently inadequate. This study was designed to evaluate the Hepatoprotective effect of saponin extract of the root of Garcinia kola on the integrity of the liver of paracetamol induced Wistar albino rats. Twenty-five male adult Wistar albino rats were divided into five (5) groups. Group I, was the Control group that received distilled water only, group II was the negative control that received 2 g/kg of paracetamol on the 13th day, and group III, IV, and V were pre-treated with 100, 200 and 400 mg/kg of the saponin extract before inducing the liver damage on the 13th day with 2 g/kg of paracetamol. Twenty-four hours after administration, the rats were sacrificed, and blood samples were collected. The serum Alanine Transaminase (ALT), Aspartate Transaminase (AST), Alkaline Phosphatase (ALP) activities, Bilirubin and Conjugated Bilirubin, Glucose and Protein concentrations were evaluated. The liver was fixed immediately in Formalin and was processed and stained with Haematoxylin and Eosin (H&E). Administration of saponin extract from the root of Garcinia kola significantly decreased paracetamol induced elevated enzymes in the test group. Also, histological observations showed that saponin extract of the root of Garcinia kola exhibited a significant liver protection against the toxicant as evident by the cells trying to return to normal. Saponin extract from the root of Garcinia kola indicated a protection of the structural integrity of the hepatocytic cell membrane and regeneration of the damaged liver.

Keywords: hepatoprotective, liver damage, Garcinia kola, saponin, paracetamol

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36 Immune Response and Histological Alteration in the Crab Carcinus aestuarii, Due to Silver Nanoparticles

Authors: Ines Kovacic, Dijana Pavicic-Hamer, Petra Buric, Maja Levak Zorinc, Daniel M. Lyons

Abstract:

Silver nanoparticles (AgNPs), owing to their unique physical and chemical properties, have become one of the most widely used nanoparticles in consumer products. Despite the increased use of AgNPs in science and industry over the past twenty years, only relatively recently has concern been raised over their entering brackish and marine environments. However, data on their potential impact on marine organisms, especially invertebrates are very limited. This study aimed to examine the effects of 60 nm AgNPs (10, 100, 500 and 1000 µg/l) and silver ions (100, 1000 µg/l) on the Mediterranean green crab Carcinus aestuarii Nardo, 1847. The crab mortality was assessed during seven days of exposure. After the exposure, total haemocytes (THC) and differential haemocytes number (DHC) were counted (immune response), in addition to histological examination of gills stained with haematoxylin and eosin. The effect of AgNPs and silver ions resulted in a dose dependent mortality and destruction of gills epithelium with haemocytes infiltration in the gills lacuna. Total haemocyte count was greater with increasing concentration of AgNPs, at concentrations from 10 to 500 µg/l. Hyalinocytes were the most common immunological cells noted in the crab hemolymph, while granulocytes and semigranulocytes were suppressed with increasing concentration of AgNPs (500 and 1000 µg/l). Thus, as crabs are filter feeders, they are susceptible to uptake of AgNPs by direct accumulation in gills mucus or indirectly via circulation of haemocytes in their open vascular system. Results of this study on crabs add to knowledge of the effects of AgNPs in the marine environment.

Keywords: crab, immune response, histological alteration, silver nanoparticles

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35 Circadian Disruption in Polycystic Ovary Syndrome Model Rats

Authors: Fangfang Wang, Fan Qu

Abstract:

Polycystic ovary syndrome (PCOS), the most common endocrinopathy among women of reproductive age, is characterized by ovarian dysfunction, hyperandrogenism and reduced fecundity. The aim of this study is to investigate whether the circadian disruption is involved in pathogenesis of PCOS in androgen-induced animal model. We established a rat model of PCOS using single subcutaneous injection with testosterone propionate on the ninth day after birth, and confirmed their PCOS-like phenotypes with vaginal smears, ovarian hematoxylin and eosin (HE) staining and serum androgen measurement. The control group rats received the vehicle only. Gene expression was detected by real-time quantitative PCR. (1) Compared with control group, PCOS model rats of 10-week group showed persistently keratinized vaginal cells, while all the control rats showed at least two consecutive estrous cycles. (2) Ovarian HE staining and histological examination showed that PCOS model rats of 10-week group presented many cystic follicles with decreased numbers of granulosa cells and corpora lutea in their ovaries, while the control rats had follicles with normal layers of granulosa cells at various stages of development and several generations of corpora lutea. (3) In the 10-week group, serum free androgen index was notably higher in PCOS model rats than controls. (4) Disturbed mRNA expression patterns of core clock genes were found in ovaries of PCOS model rats of 10-week group. Abnormal expression of key genes associated with circadian rhythm in ovary may be one of the mechanisms for ovarian dysfunction in PCOS model rats induced by androgen.

Keywords: polycystic ovary syndrome, androgen, animal model, circadian disruption

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34 In vitro Effects of Salvia officinalis on Bovine Spermatozoa

Authors: Eva Tvrdá, Boris Botman, Marek Halenár, Tomáš Slanina, Norbert Lukáč

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In vitro storage and processing of animal semen represents a risk factor to spermatozoa vitality, potentially leading to reduced fertility. A variety of substances isolated from natural sources may exhibit protective or antioxidant properties on the spermatozoon, thus extending the lifespan of stored ejaculates. This study compared the ability of different concentrations of the Salvia officinalis extract on the motility, mitochondrial activity, viability and reactive oxygen species (ROS) production by bovine spermatozoa during different time periods (0, 2, 6 and 24 h) of in vitro culture. Spermatozoa motility was assessed using the Computer-assisted sperm analysis (CASA) system. Cell viability was examined using the metabolic activity MTT assay, the eosin-nigrosin staining technique was used to evaluate the sperm viability and ROS generation was quantified using luminometry. The CASA analysis revealed that the motility in the experimental groups supplemented with 0.5-2 µg/mL Salvia extract was significantly lower in comparison with the control (P<0.05; Time 24 h). At the same time, a long-term exposure of spermatozoa to concentrations ranging between 0.05 µg/mL and 2 µg/mL had a negative impact on the mitochondrial metabolism (P<0.05; Time 24 h). The viability staining revealed that 0.001-1 µg/mL Salvia extract had no effects on bovine male gametes, however 2 µg/mL Salvia had a persisting negative effect on spermatozoa (P<0.05). Furthermore 0.05-2 µg/mL Salvia exhibited an immediate ROS-promoting effect on the sperm culture (P>0.05; Time 0 h and 2 h), which remained significant throughout the entire in vitro culture (P<0.05; Time 24 h). Our results point out to the necessity to examine specific effects the biomolecules present in Salvia officinalis may have individually or collectively on the in vitro sperm vitality and oxidative profile.

Keywords: bulls, CASA, MTT test, reactive oxygen species, sage, Salvia officinalis, spermatozoa

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33 Evaluation of Anti-Typhoid Effects of Azadirachta indica L. Fractions

Authors: A. Adetutu, T. M. Awodugba, O. A. Owoade

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The development of resistance to currently known conventional anti-typhoid drugs has necessitated search into cheap, more potent and less toxic anti-typhoid drugs of plant origin. Therefore, this study investigated the anti-typhoid activity of fractions of A. indica in Salmonella typhi infected rats. Leaves of A. indica were extracted in methanol and fractionated into n-hexane, chloroform, ethyl-acetate, and aqueous fractions. The anti-salmonella potentials of fractions of A. indica were assessed via in-vitro inhibition of S. typhi using agar well diffusion, Minimum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC) and biofilm assays. The biochemical and haematological parameters were determined by spectrophotometric methods. The histological analysis was performed using Haematoxylin and Eosin staining methods. Data analysis was performed by one-way ANOVA. Results of this study showed that S. typhi was sensitive to aqueous and chloroform fractions of A. indica, and the fractions showed biofilm inhibition at concentrations of 12.50, 1.562, and 0.39 mg/mL. In the in-vivo study, the extract and chloroform fraction had significant (p < 0.05) effects on the number of viable S. typhi recovered from the blood and stopped salmonellosis after 6 days of treatment of rats at 500 mg/kg b.w. Treatments of infected rats with chloroform and aqueous fractions of A. indica normalized the haematological parameters in the animals. Similarly, treatment with fractions of the plants sustained a normal antioxidant status when compared with the normal control group. Chloroform and ethyl-acetate fractions of A. indica reversed the liver and intestinal degeneration induced by S. typhi infection in rats. The present investigation indicated that the aqueous and chloroform fractions of A. indica showed the potential to provide an effective treatment for salmonellosis, including typhoid fever. The results of the study may justify the ethno-medicinal use of the extract in traditional medicine for the treatment of typhoid and salmonella infections.

Keywords: Azadirachta indica L, salmonella, typhoid, leave fractions

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