Haematological and histopathological effects of apigenin, phloretin and myricetin based on uterotrophic assay in immature Wistar female albino rats

2014 ◽  
Vol 34 (7) ◽  
pp. 755-768
Author(s):  
N Barlas ◽  
G Karabulut
Keyword(s):  
Ethinyl Estradiol ◽  
Negative Control Group ◽  
Positive Control ◽  
Negative Control ◽  
Female Rats ◽  
Control Group ◽  
Albino Rats ◽  
Immature Female ◽  
Tissue Samples ◽  
Uterotrophic Assay

In this study, it is aimed to determine the histopathological and haematological effects of apigenin, phloretin and myricetin on Wistar immature female rats using Tier 2 of the uterotrophic assay. The female rats were divided into 17 groups with 6 rats in each group. There was a negative control group and positive control dose groups that contained 0.07 µg/kg/day, 0.7 µg/kg/day and 7 µg/kg/day of ethinyl estradiol (EE), 0.7 µg/kg/day 17α-ethinyl estradiol + 1 mg/kg/day tamoxifen and genistein. The other dose groups contain 1 mg/kg/day, 10 mg/kg/day and 100 mg/kg/day of apigenin, myricetin and phloretin. All chemicals had been given to Wistar immature female rats with oral gavage for three consecutive days. At the end of the study, blood samples were analysed for haematological parameters. Tissue samples that were taken from the liver, kidney, spleen and thyroid were histopathologically and histomorphometrically examined. There were no significant differences between oil control and other dose groups for glomerular histomorphometry. However, there were siginificant differences for thyroid histomorphometry. Especially, 10 and 100 mg/kg/day of phloretin dose groups had a siginificant increase in colloid surface area in thyroid compared with the 1 mg/kg/day of phloretin and oil control groups. Significant histopathological changes (congestion, degeneration, fibrosis and mononuclear cell infiltration) were noted in the tissue specimens obtained from the treatment groups compared with the control group. According to the results of the haematological analysis of the groups, especially the values of erythrocytes and haematocrit were increased significantly in most of the dose groups according to the oil control group.

scholarly journals Effects of avilamycin and essential oils on mRNA expression of apoptotic and inflammatory markers and gut morphology of piglets

2008 ◽  
Vol 53 (No. 9) ◽  
pp. 377-387 ◽  
Author(s):  
A. Kroismayr ◽  
J. Sehm ◽  
M.W. Pfaffl ◽  
K. Schedle ◽  
C. Plitzner ◽  
...  
Keyword(s):  
Essential Oils ◽  
Negative Control Group ◽  
Positive Control ◽  
Negative Control ◽  
Feed Additives ◽  
Feed Additive ◽  
Pcr Analysis ◽  
Control Group ◽  
Mesenteric Lymph Nodes ◽  
Tissue Samples

In this study 120 piglets were allotted to 3 dietary treatments, negative control group, one group receiving a blend of essential oils (EO) derived from oregano, anise and citrus peels (40 mg/kg diet), and a positive control group treated with avilamycin as growth promoting feed additive (40 mg/kg diet). On day 22 of the experiment, 12 representative animals from each treatment group were sacrificed and tissue samples were collected for quantitative real time-PCR analysis and gut tissue morphology. EO and avilamycin decreased the gene expression of the transcriptional factor NFκB and the apoptotic marker TNFα significantly in the ileum and jejunum, respectively. The expression of the proliferation marker Cyclin D1 was also significantly decreased by both substances in the colon, mesenteric lymph nodes and spleen. The colonic crypt depth was reduced by avilamycin, and also numerically by the essential oils. These changes correlated with the up-regulation of the apoptosis factor Caspase 3. Histomorphometry revealed a smaller size of ileal Peyer’s patches through the use of both feed additives, which correlated significantly with lower expression rates of NFκB. In conclusion, the results suggest that EO and avilamycin relieved weaning piglets from an immune defence stress in a similar way.

scholarly journals Anti-Oxidant and Hepatoprotective Effects of Senecio biafrae on CCl4-induced Liver Damage in Rats

2019 ◽  
Vol 13 (2) ◽  
pp. 31-35
Author(s):  
Israel Oghenevwodoko Okoro ◽  
◽  
Helen Ejiro Kadiri ◽  
Keyword(s):  
Negative Control Group ◽  
Positive Control ◽  
Negative Control ◽  
Control Group ◽  
Albino Rats ◽  
Root Extract ◽  
Distilled Water ◽  
Standard Drug ◽  
Anti Oxidant

Background: The present study was performed to explore whether the aqueous extract of Senecio biafrae (S. biafrae) roots provide any in vivo protective activity against carbon tetrachloride (CCl4)-induced hepatotoxicity in male albino rats. Methods: Rats (150-200 grams) were grouped into five groups (A-E) of six rats each and were treated orally for twelve days with 72 hourly administration of CCl4 (1 mL/kg) as follows: Group A received distilled water only (negative control), Group B was administered distilled water plus CCl4 (positive control), Group C was administered 400 mg/kg extract and CCl4, Group D received 200 mg/extract and CCl4, while Group E was administered standard drug (Silymarin 25mg/kg, PO). Results: Pre-treatment with the extract of S. biafrae (200 or 400mg/kg) or Silymarin (25mg/kg) caused significant restoration in the biomarkers as evaluated by reducing the levels of malondialdehyde, transaminases and elevating the levels of superoxide dismutase, catalase and glutathione peroxidase activities, which were altered by CCl4 toxicity. The extract at a dose of 400mg/kg demonstrated similar activities comparable to the standard drug (Silymarin). Conclusion: The results of this study indicate that the root extract of S. biafrae possesses hepatoprotective and anti-oxidant properties which may be due to the presence of phytochemicals in it.

scholarly journals Comparative Study of the Protective Effect of Cola anomala and Coffea arabica Against Induced Toxicity in Rats

2020 ◽  
Vol 9 (5) ◽  
pp. 1
Author(s):  
M-A. Angie Mbong ◽  
F. L. Ebouel Edoun ◽  
L. C. Manga Ngandi ◽  
J. A. Fotso Youvop ◽  
R. Orang Orang ◽  
...  
Keyword(s):  
Lipid Profile ◽  
Metabolic Disorders ◽  
Liver Toxicity ◽  
Hdl Cholesterol ◽  
Negative Control Group ◽  
Positive Control ◽  
Negative Control ◽  
Oxidative Status ◽  
Female Rats ◽  
Control Group

This work was aimed at evaluating the effects of C. anomala and C. arabica on Methotrexate (MTX) induced metabolic disorders. For this, the aqueous extract (AE) of C. anomala and C. arabica were prepared and their polyphenols, flavonoids and alkaloids contents determined as well as their antiradical and total antioxidant capacity. An animal experimentation using female rats was carried out for 14 days. Rats were divided into 6 groups; a negative control group receiving water; a positive control group receiving 12mg/Kg Bw of MTX; four tests groups receiving 12mg/Kg Bw of MTX and one of the extracts at a dose of 200mg/Kg Bw or 400mg/Kg Bw. At the end of the experiment, plasmas and hemolysates were prepared as well as liver and kidney homogenates for the evaluation of oxidative status (catalase, total protein and malondialdehyde (MDA)), liver toxicity (alanine amino transferase (ALT)) and renal toxicity (creatinine and urea) and lipid profile (triglycerides, total cholesterol and HDL-cholesterol). Weight gain in extract-treated rats was better with the C. anomala. Concerning oxidative status, MDA levels were generally lower in C. anomala-treated groups compared to C. arabica while catalase level was higher in C. anomala-treated rats. As for lipid profile, it is C. arabica that showed more or less better results. Both extracts led to an amelioration of toxicity markers compared to exclusive treatment with MTX. The results of this study suggest that C. anomala and C. arabica may reduce metabolic disorders associated with the intake of MTX during cancer treatment; C. anomala protecting better C. arabica.

The Effect of Herbal Infusion on Fasting Blood Glucose (FBS) and Malondyaldehyde (MDA) Levels in Diabetic Rats

2020 ◽  
Vol 1009 ◽  
pp. 9-14
Author(s):  
Apriliani Ismi Fauziah ◽  
Lintang Dian Saraswati ◽  
Ari Udiyono ◽  
Beti Safitri
Keyword(s):  
Blood Glucose ◽  
Fasting Blood Glucose ◽  
Aloe Vera ◽  
Control Sample ◽  
Negative Control Group ◽  
Positive Control ◽  
Negative Control ◽  
Diabetic Rats ◽  
Control Group ◽  
Albino Rats

Aloe vera, ginger, and sappan wood have recently gained attention as a potent antioxidant. This study was undertaken to analyze the possible effects of oral administration of herbal infusion (the combination of Aloe vera, ginger, and sapang wood) on fasting blood glucose (FBS) and malondialdehyde (MDA) levels in diabetic rats. Forty male albino rats (150-250 gm) were divided into five groups (n=8 in each group): alloxan-induced diabetic (negative control group/K-), acarbose treated (positive control group/K+), diabetic rats treated with herbal infusion dose 6.75 mg/150 gmbw (P1), diabetic rats treated with herbal infusion dose 13.5 mg/150 gmbw (P2), and diabetic rats treated with herbal infusion dose 20.25 mg/150 gmbw (P3). Male albino rats induced with a dose of alloxan 120 mg/kgbw and also 5% glucose after 6 hours of alloxan induction. FBS and MDA levels of each rat were measured before induction of alloxan was performed as a control sample. After seven days and 14 days of herbal infusion administration, FBS parameters was investigated with a glucometer. In the last week, the concentration of MDA in serum blood was determined using a UV-VIS spectrophotometer with a wavelength of 545 nm. Diabetic rats exhibited a significant decrease in FBS and MDA. Administration of herbal infusion was reduced FBS significantly between groups (ANOVA, Bonferroni, p<0.05). The concentration of MDA in serum blood was decreased significantly on K-, K+, P1, P2, P3 compared to the control group (ANOVA, LSD, P<0.05). It could be postulated herbal infusion has anti-hyperglycemic and anti-diabetic effects of FBS and MDA in male albino rats Wistar strain.

scholarly journals Evaluation of Protective Properties of Elaeis oleifera Fruit Extract on Renal Parameters of Dichlorvos-Induced Nephrotoxocity in Albino Rats

2020 ◽  
pp. 1-13
Author(s):  
Edna Ogechi Nwachuku ◽  
Fedelis Beega ◽  
Adline Erinma Ben- Chioma ◽  
Ngozi Brisibe ◽  
Ibioku Elekima
Keyword(s):  
Treatment Group ◽  
Palm Oil ◽  
Negative Control Group ◽  
Positive Control ◽  
Negative Control ◽  
Positive Control Group ◽  
Control Group ◽  
Albino Rats ◽  
Study Group ◽  
Acute Study

Aim: Evaluate the protective effects of palm oil on renal parameters after dichlorvos toxicity in albino rats. Study Design and Methodology: The study consisted of 3 phases: The acute study which lasted for 24 hours, the sub-acute study which lasted for 14 days and the sub chronic study which lasted for 30 days. The design and treatment pattern is shown below. Phase 1: Acute Study. Group 1: No DDVP, No palm oil for 24 hours (Negative control), Group 2: 30 mg/kg of DDVP without palm oil (positive control), Group 3: 30 mg/kg of DDVP and 100 mg/kg palm oil for 24 hours (treatment group). Phase 2: Sub-Acute (14 days) Study. Group 4: No DDVP, No palm oil for 14 days (Negative control), Group 5: 10 mg/kg of DDVP without palm oil daily for 14 days (positive control), Group 6: 10 mg/kg of DDVP and 100 mg/kg of palm oil daily for 14 days (positive control). Phase 3: Sub-Chronic (30 days) Study. Group 7: No DDVP, No palm oil for 30 days (Negative control), Group 8: 10 mg/kg of DDVP without palm oil daily for 30 days (positive control), Group 9: 10 mg/kg of DDVP and 100 mg/kg palm oil daily for 30 days (treatment group). All administration was done orally. After the period of treatments, the rats were sacrificed after 18 hours of fast. Whole blood samples (5 mls) were collected into lithium heparin bottle and spun at 3500 rpm for 5 minutes to obtain plasma samples. Samples obtained were used for the determination of Na+, K+, HCO3, urea, and creatinine while renal tissues obtained were used for histopathological examinations. Results: Significantly higher values were seen in urea in the dichlorvos treated rats over a period of 24 hours, 14 days, and 30 days as compared to rats co-treated with palm oil and the control. Creatinine indicated significantly higher over a period of 24 hours while non-significant increases were observed in the dichlorvos treated rats over a period of 14 days and 30 days. More so, significantly higher values were seen in potassium in the dichlorvos treated rats over a period of 24 hours and 14 days, while significantly higher values in potassium were seen after period of 30 days as compared to rats co-treated with palm oil and the control. Sodium and chloride did not indicate significant difference over the period of 24 hours, 14 days, and 30 days. Histological examination of the renal tissue indicated structural distortions dichlorvos treated rats over a period of 24 hours, 14 days and 30 days while significant improvements in the structural integrity of the kidney were observed in rats co-treated with palm oil. Conclusion: Results obtained indicated that palm oil showed a protective effect in ameliorating the nephrotoxicity induced by dichlorvos as shown by the histological examination and decreased values of creatinine and urea as well as potassium in palm oil treated rats.

scholarly journals Effect of Chitosan on Histology of Reproductive Organs of Female Wistar Rats (Rattus norvegicus) Exposed to Acetate Lead

2019 ◽  
Vol 30 (4) ◽  
pp. 259
Author(s):  
Alief Ayu Purwitasari ◽  
Annisa' Wigati Rozifa ◽  
Dwi Dianita Irawan ◽  
Umi Kalsum ◽  
Retty Ratnawati ◽  
...  
Keyword(s):  
Treatment Group ◽  
Lead Acetate ◽  
Antral Follicle ◽  
Negative Control Group ◽  
Positive Control ◽  
Negative Control ◽  
Female Rats ◽  
Control Group ◽  
P Value ◽  
Control Group Design

Lead is one of the pollutants widely spread in the environment because it is not easily decomposed. Lead can affect system functions such as the ovary and endometrium. Lead can trigger oxidative stress by reducing antioxidant enzymes and increasing Reactive Oxygen Species (ROS). Lead can also reduce Follicle Stimulating Hormone (FSH) and Luteinizing Hormone (LH) levels by disturbing the hypothalamus. Chitosan is an antioxidant compound that can reduce the toxic effects of lead. The purpose of this study was to study the effects of chitosan administration on the diameter of antral follicles, the number of endometrial arterioles, and the thickness of endometrial rats after lead acetate exposure. This study was an experimental laboratory using a posttest-only control group design approach applied on 25 female rats aged 8 weeks old, body weight 125-175 grams. Lead and chitosan were given orally with a sonde. There were 5 groups, namely, negative control group (without any treatment), positive control group (lead 175mg/kg/BW), treatment group 1 (lead 175mg/kg/BW + chitosan 16mg/kg/BW, treatment group 2 (lead 175mg/kg/BW + chitosan 32mg/kg/BW), and treatment group 3 (lead 175mg/kg/BW + chitosan 64mg/kg/BW) for 30 days. The rats were sacrificed at proestrus phase, which was proven from vaginal swab. Observations were carried out using the Hematoxylin Eosin (HE) staining method. The observations were analyzed using One Way ANOVA and followed by Least Significant Differences (LSD) test. The results showed significant results (p-value &lt;0.05). Chitosan can increase the diameter of the antral follicle, increase the number of endometrial arterioles, and increase the thickness of endometrial rats exposed by lead acetate.

scholarly journals Anti-Anaemic Effect of Cnidoscolus chayamansa (Mc Vaugh) Leaf Extract on Phenyl Hydrazine-Treated Rats

2020 ◽  
pp. 23-32
Author(s):  
R. J. Kutshik ◽  
M. K. Jiyil ◽  
C. E. Mafuyai ◽  
L. L. Ichor
Keyword(s):  
Leaf Extract ◽  
Negative Control Group ◽  
Positive Control ◽  
Negative Control ◽  
Untreated Group ◽  
Control Group ◽  
Albino Rats ◽  
Phenylhydrazine Hydrochloride ◽  
Blood Disorder ◽  
Cnidoscolus Chayamansa

Background: Anaemia is one of the world's threatening disease conditions of blood disorder that leads to the decrease in red blood cells which affects people of all ages. Aim of the Study: This study aimed to evaluate the effects of Cnidoscolus chayamansa leaf extract on phenylhydrazine hydrochloride induced anaemia in albino rats. Study Duration: This study was conducted on 30th June 2019 at the Department of Biochemistry, Faculty of Basic Medical Sciences, University of Jos, Nigeria. Methodology: A total of 16 rats were split into four groups. Group A the baseline for negative control, group B positive control, group C pre-treatment and group D post-treatment. Induction was done by intraperitoneal with phenylhydrazine at a dose of 40 mg/kg. Blood was taken for each 2 days interval for PCV, RBC, WBC and Hb. Preliminary phytochemical screening was investigated by standard procedures. Results: The results show a significant (P<0.05) increase in serum PCV, RBC, WBC and Hb concentration when compared with the untreated group. Decrease in serum alanine aminotransferase (15.46±4.443 & 7.80±0.429), aspartate aminotransferase (20.89±1.095 & 11.66±0.898), total bilirubin (13.70±0.351 & 10.45±0.620), direct bilirubin (7.49±0.659 & 3.76±0.042) and alkaline phosphatase (254.25±2.287 & 192.00±1.474) and a significant (P<0.05) increase in serum protein and albumin when compared with untreated group. Iron binding shows a significantly (P<0.05) increase (153.49±0.530 & 166.09±1.334) and (85.16±0.824 &98.43±0.562) while there was a significant (p<0.05) decrease in serum Urea (2.39±0.027&4.70±0.047), Creatinine (68.38±0.404&75.01±0.891), and Uric acid (234.92±2.761&246.52±1.136) when compared with the baselines.  Conclusion: The positive effect of the vegetable may be attributed to its rich phytochemicals, nutrients which supports the use of the leaves for food and ethnomedicinal purposes in many parts of Nigeria.

The Comparison of Ostecyte in the Pressure area and Tension area on Tooth Movement Because of Hyperbaric Oxygen Therapy

DENTA ◽  
2018 ◽  
Vol 12 (1) ◽  
pp. 34
Author(s):  
Arya Barahmanta ◽  
Muhammad Faizal Winaris ◽  
Pambudi Raharjo
Keyword(s):  
Hyperbaric Oxygen ◽  
Oxygen Therapy ◽  
Tooth Movement ◽  
Bone Remodelling ◽  
Orthodontic Tooth Movement ◽  
Negative Control Group ◽  
Positive Control ◽  
Negative Control ◽  
Control Group ◽  
Pressure Area

<p><strong><em>Background:</em></strong><em> Orthodontic tooth movement is a </em><em>interaction prosess</em><em> of resorption and deposition of bone remodeling. Orthodontic tooth movement by mechanical strength causes changes in alveolar bone. Osteocyte is an essential cell to respond bone remodelling. Hyperbaric Oxygen Therapy affects production of osteocyte because it can release Reactive Oxygen Species (ROS) and Nitrid Oxide (NO).  <strong>Purpose: </strong>To determine the difference number  of osteocyte in pressure and tension area during tooth movement by adjuvant of Hyperbaric Oxygen 2,4 ATA during 7 days starting on day 8 to day 14. <strong>Materials and Methods</strong>: This research used Completery Randomized Control Group Post Test Only Design. 36 cavia cobaya (male)  were divided into 3 groups randomly : the negative control groups, positive control group, and treatment group. Preparat staining used Hematoxylin Eosin (HE) and calculated on microscop 1000x with 20 field of view. Data analyses used one way ANOVA and LSD test then compared each area by using paired T test. <strong>Result:</strong> The data showed that the treatment group (P=10,67) tension area has the highest number of osteocyte than  negative control group (K-=3,67), positive control (K+=7,42). In the pressure area showed that negative control group (K-=5,00) has the highest  than positive control group (K+=3,83) and treatment (P=3,25). <strong>Conclusion: </strong>Therapy HBO 2,4 ATA 7 days starting on day 8 to day 14 is could increase osteocyte in the tissue to stimulate process of bone remodelling.</em></p><pre><strong> </strong></pre><p><strong><em>Keywords:</em></strong><em> Hyperbaric Oxygen, Tooth movement, Bone remodeling, </em><em>Osteocyte</em><em></em></p><p><em> </em></p><p><strong><em>Correspondence:</em></strong><em> </em><em>Arya Brahmanta</em><em>, Department of Orthodonty, Faculty of Dentistry, Hang Tuah University, Arif Rahman Hakim 150, Surabaya, Phone 031-5945864, Email:</em><em> </em><a href="mailto:[email protected]"><em>arya.brahmanta</em><em>@</em><em>hangtuah.ac.id</em></a></p>

Efek Ekstrak Etanol Kulit Petai (Parkia speciosa Hassk) Terhadap Penurunan Kadar Glukosa Darah Mencit Jantan

2018 ◽  
Vol 3 (1) ◽  
pp. 1
Author(s):  
Verawaty Verawaty ◽  
Dhea Claudia Novel
Keyword(s):  
Blood Glucose ◽  
Ethanol Extract ◽  
Negative Control Group ◽  
Positive Control ◽  
Negative Control ◽  
Control Group ◽  
Male Mice ◽  
Glucose Levels ◽  
The Third ◽  
Blood Glucose Levels

<p>Penelitian ini bertujuan untuk melihat pengaruh pemberian ekstrak etanol kulit petai (Parkia speciosa Hassk) terhadap penurunan kadar glukosa darah mencit jantan yang diinduksi aloksan. Hewan percobaan dibagi atas 5 kelompok diantaranya kelompok kontrol negatif, kelompok kontrol positif,dosis I (280 mg/kgBB mencit), dosis II (560 mg/kg BB mencit), dosis III (840 mg/kg BB mencit). Penelitian dilakukan selama 21 hari. Persentase penurunan kadar glukosa darah mencit jantan setelah diberikan ekstrak etanol kulit petai pada hari ke-21 adalah dosis I (77,52 %) lebih besar dibandingkan dengan dosis II (69,5 %) dan dosis III (73,37 %). Data yang diperoleh dianalisis dengan uji Two Way Anova dengan program SPSS 17. Hasil penelitian ini menunjukkan bahwa pemberian ekstrak etanol kulit petai untuk tiga variasi dosis menyatakan perbedaan yang bermakna secara statistik terhadap penurunan kadar glukosa darah mencit jantan.</p><p><em>Petai (Parkia speciosa Hassk) has a compound β-sitosterol and stigmasterol that have efficacy to decreased blood glucose levels. This study aimed to determine the effect of ethanol extract of petai peel for decrease blood glucose levels of male mice induced by alloxan. Experimental animals were divided into 5 groups including negative control group, positive control group, the first dose (280 mg/kg in mice), the second dose (560 mg/kg in mice), the third dose (840 mg/kg in mice). The study was conducted for 21 days. After 21 days, the result found that the percentage of blood glucose levels after the male mice given the ethanol extract of petai peel was, the first dose (77.52%) biger than the second dose (69.5%) and the third dose (73.37%). The data obtained were analyzed by Two Way ANOVA using SPSS 17. The results showed that have signicantly difference between three dose variation of ethanol extract of petai peel in blood glucose levels.</em></p>

scholarly journals Resistance to Fracture of Dental Roots Obturated with Different Materials

2015 ◽  
Vol 2015 ◽  
pp. 1-5 ◽  
Author(s):  
Berkan Celikten ◽  
Ceren Feriha Uzuntas ◽  
Kamran Gulsahi
Keyword(s):  
Fracture Resistance ◽  
Testing Machine ◽  
Negative Control Group ◽  
Positive Control ◽  
Negative Control ◽  
Positive Control Group ◽  
Control Group ◽  
Root Canal Filling ◽  
Filling Materials ◽  
Group 2

The aim of this study was to compare the vertical fracture resistance of roots obturated with different root canal filling materials and sealers. Crowns of 55 extracted mandibular premolar teeth were removed to provide root lengths of 13 mm. Five roots were saved as negative control group (canals unprepared and unfilled). Fifty root canals were instrumented and then five roots were saved as positive control group (canals prepared but unfilled). The remaining 45 roots were randomly divided into three experimental groups (n=15root/group) and obturated with the following procedures: in group 1, glass ionomer-based sealer and cone (ActiV GP obturation system); in group 2, bioceramic sealer and cone (EndoSequence BC obturation system); and in group 3, roots were filled with bioceramic sealer and cone (Smartpaste bio obturation system). All specimens were tested in a universal testing machine for measuring fracture resistance. For each root, the force at the time of fracture was recorded in Newtons. The statistical analysis was performed by using Kruskal-Wallis and post hoc test. There were no significant differences between the three experimental groups. The fracture values of three experimental and negative control groups were significantly higher than the positive control group. Within the limitations of this study, all materials increased the fracture resistance of instrumented roots.

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