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ORIGINAL ARTICLES
Volatile compounds, antioxidants, and anticancer activities of Cape gooseberry fruit (
Physalis peruviana
L.): An
in-vitro
study
Manal M Ramadan, Ahmed H El-Ghorab, Kadry Z Ghanem
July-December 2015, 10(2):56-64
DOI
:10.4103/1687-4293.175556
Background/aim
Cape gooseberry is golden-colored spherical fruit commercially produced in Egypt. It is primarily used in folk medicine for treating some diseases. To identify the aroma compounds in Cape gooseberry and to evaluate its antioxidant activities as well as its anticancer (for colon and breast cancers) effects in human cell lines.
Materials and methods
The volatile compounds were identified using gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS). Polyphenols (phenolics and flavonoids) were also determined. Antioxidant activity was determined by three different methods: 2,2΄-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azinobis(3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS), and ferric reducing antioxidant power (FRAP) assays. Anticancer (for colon or breast cancer) activity was determined in cancer cell lines using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay.
Results
A total of 34 components of the essential oil were identified by GC and GC-MS. The volatile compounds were grouped in classes of substances, including 11 terpene compounds (six monoterpenoids and five sesquiterpene), 11 esters, five alcohols, two phenolic compounds, two aldehydes, two ketones, and one lactone. Terpenes (monoterpenes and sesquiterpenes) were the most abundant volatile constituents, accounting for the largest portion of the total volatiles (36.09%). The next most abundant compounds were esters, comprising 17.17% of the total volatile components identified. Phenolic compounds were the next most abundant compounds, comprising 16.04% of the total volatiles. Alcohols and aldehydes represented 6.37 and 1.88% of the total volatile compounds, respectively. Ketones and lactones are less abundant in the profile of volatile compounds in Cape gooseberry. Ethanol extract had higher phenolic and flavonoid contents than did hexane extract. As ethanol extract of Cape gooseberry achieved higher antioxidant activity than did hexane extract, it tested as an anticancer (for colon or breast cancer) agent. Cape gooseberry extract was more potent in inhibiting colon cell lines (IC
50
: 142 μg/ml) compared with breast cell line (IC
50
: 371 μg/ml).
Conclusion
Egyptian Cape gooseberry fruits may be suggested as a potential source of natural antioxidants and anticancer agents.
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13
4,659
615
Volatile compounds and antioxidant activity of the aromatic herb
Anethum graveolens
Manal M Ramadan, Nadia N Abd-Algader, Hatil H El-kamali, Kadry Z Ghanem, Abdel Razik H Farrag
July-December 2013, 8(2):79-88
DOI
:10.4103/1687-4293.123791
Background/Aim
Anethum graveolens
L. (dill) (Apiaceae) is one of the most popular culinary herbs in the world. Dill has been cultivated since ancient times, and the use of this plant for medicinal and consumption purposes has been recorded dating back to the Greek and Egyptian civilizations. Dill is widely used to give flavor to food. The aim of this work was to assess the chemical composition of the volatile compounds in dill as well as their hepatoprotective and nephroprotective activity against free radicals generated by paracetamol.
Materials and methods
The chemical composition of the volatile compounds of dill was assessed by gas chromatography and gas chromatography-mass spectrometry. Four groups of rats were used (the normal control group, the
A.
graveolens-
supplemented group, the paracetamol-intoxicated group, and the
A.
graveolens-
protected group) for studying the effect of plant infusion on panadol (paracetamol)-induced free radicals and hepatotoxicity. Plasma total antioxidant capacity, plasma catalase, cellular glutathione peroxidase, plasma total protein, albumin, alanine amino transferase, aspartate amino transferase, alkaline phosphate, γ-glutamyl transferase, total bilirubin, and direct bilirubin levels were determined. In addition, kidney functions (plasma urea and creatinine) and histopathological and histochemical changes in the liver were investigated.
Results
The phytochemical results identified volatile components 7-α-hydroxy manool (24.43%), l-carvone (14.28%), limonene (13.9%), epi-α-bisabolol (6.81%), α-terpinene (5.44%), and α-phellandrene (4.63%) as the main constituents.
p
-Cymene (2.13%), sabinene (1.98), and α-pinene (1.43%) were determined as the minor constituents. The biochemical results showed that a mega dose of paracetamol induced the production of free radicals, which caused damage to hepatocytes and nephrocytes in rats. The aqueous extract of dill revealed high antioxidant properties and acted as an extracellular neutralizer of free radicals. Histopathological and histochemical observations showed severe damage in the liver. Supplementation with dill in paracetamol-intoxicated rats attenuated the damage to the liver.
Conclusion
The present study revealed that
A. graveolens
has antihepatotoxic properties that could minimize the deleterious effects generated by hepatotoxin paracetamol, and therefore it can be used as a potent antihepatotoxic agent.
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270
Serum vitamin D level as a predictor of bronchial asthma in Egyptian children
Hala G Elnadya, Eman M Foudac, Ola M Elsheikha, Inas R ElAlameeya, Amal I Elshafied, Lobna S Sherifa, Eman R Younessb, Nadia A Mohamedb
July-December 2013, 8(2):67-73
DOI
:10.4103/1687-4293.123788
Background/Aim
This study was designed to assess the relationship between the serum level of 25-hydroxyvitamin D and the clinical, functional severity and the level of asthma control among Egyptian asthmatic children.
Patients and methods
This case-control cross-sectional study was conducted on 50 asthmatic patients from those regularly attending the Pediatric Chest Clinic, Children's Hospital, Ain Shams University. Twenty healthy children of matched age and sex were recruited as the control group.
Results
Pulmonary function tests were significantly decreased in the asthmatic cases compared with the control (
P
< 0.01). The serum vitamin D level was found to be significantly decreased in asthmatic children compared with the control group (
P
< 0.001). Out of the 50 asthmatic children, 20 had a serum vitamin D level of less than 25 ng/ml, and were considered to be 'vitamin D deficient', whereas the remaining 30 children had a level ranging between >25 ng/ml and ≤30 ng/ml and were considered to be 'vitamin D insufficient'. The asthmatic children had a highly significant increase in the total leukocytic count, the eosinophilic count, serum immunoglobulin E, and serum alkaline phosphatase (
P
< 0.001 in all). There was also a significant increase in the serum phosphorus level in the asthmatic children group compared with the control group (
P
< 0.01). The serum vitamin D level was 24.1 ng/ml ± 2.9 in the asthmatic patients on inhaled corticosteroid therapy and 28 ng/ml ± 1.4 in those not on inhaled corticosteroid therapy. Significant positive correlations were found between the serum vitamin D level and the predicted percentage of forced expiratory volume in 1 s (
r
= 0.871,
P
< 0.001). A highly significant negative correlation was found between the serum vitamin D level and serum immunoglobulin E (marker of allergy;
r
= −0.589,
P
< 0.001). Moreover, serum vitamin D showed an inverse correlation with the clinical severity of bronchial asthma (
r
= 0.903,
P
< 0.001) and the level of control of asthma (
r
= 0.923,
P
< 0.001), classified according to the Global Initiative for Asthma (GINA) classification. The accuracy of vitamin D as a predictor of asthma was found to be 88% by automatic linear modeling.
Conclusion
Our results revealed an important support for the association between bronchial asthma in children and vitamin D deficiency, with a direct relationship between its serum level and pulmonary function test measures and the increased asthma severity. Vitamin D deficiency can be considered as a strong predictor of asthma. Improving the vitamin D status can help in the primary prevention of asthma and in decreasing exacerbations of attacks. Clinical trials of vitamin D supplementation to prevent asthma exacerbation are recommended.
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1,780
149
Evaluation of the phenolic content and antioxidant potential of
Althaea rosea
cultivated in Egypt
Nagwa M Ammar, El-Sayeda A El-Kashoury, Lamiaa T Abou El-Kassem, Rasha E Abd El-Hakeem
July-December 2013, 8(2):48-52
DOI
:10.4103/1687-4293.123786
Objective
The aim of this study was to evaluate the total phenolic content (TPC) and antioxidant potential of
Althaea rosea
Cav., family Malvaceae, as well as to isolate and identify the flavonoid content of the methanolic extract of the aerial parts. In addition, a comparison between the TPC and antioxidant capacity of the methanolic extract of both aerial parts and flowers was carried out to discover new active constituents that can be utilized in drug industry.
Materials and methods
The extraction of the flavonoid compounds was carried out by percolation of the dried aerial parts of the plant under investigation with 70% methanol until exhaustion. The combined extract was then concentrated and defatted with petroleum ether (60-80°C). After separation of the lipoidal matter, the remaining extract was purified from mucilage and subjected to several column chromatographic techniques for isolation of the flavonoids. The identification of flavonoid compounds was carried out using physical, chemical, and spectral methods such as ultraviolet,
1
H NMR, and
13
C NMR. The antioxidant potential of the methanolic extracts of both aerial parts and flowers was determined using the s[table 2], 2-diphenyl-1-picrylhydrazyl free radical scavenging activity method. Furthermore, their TPC was also determined using the Folin-Ciocalteu method.
Results
Five flavonoid compounds were isolated from the aerial parts of
A. rosea
Cav., which were identified as quercetin 3-
O
-β-d-glucuronopyranoside-8-C-β-d-glucopyranoside, kaempherol-3-
O
-β-d-rutinoside, kaempherol-4΄-
O
-β-d-glucoside, kaempherol-3-
O
-β-d-glucoside, and kaempherol. The antioxidant activity was measured in terms of their IC
50
. The IC
50
values of the methanolic extracts of the aerial parts and flowers were 11 and 1 mg/ml, respectively, whereas the TPCs were 48 and 73 μg/ml, respectively.
Conclusion
The methanolic extracts of both aerial parts and flowers of
A. rosea
Cav. are rich in phenolic compounds and have a prominent antioxidant activity. The antioxidant activity of both extracts may be attributed to their phenolic content.
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2,136
254
Health related quality of life of children with epilepsy in Egypt
Zeinab M Monir, Inas R EL-Alameey, Eman Eltahlawy
July-December 2013, 8(2):53-66
DOI
:10.4103/1687-4293.123787
Background/aim
Children with epilepsy are at an increased risk of poor health-related quality of life even in the absence of active seizures. The study was undertaken to assess the health-related quality of life and its predictors in children with epilepsy, comparing the relationship between different types of seizures and health-related quality of life in Egypt.
Patients/methods
This cross sectional case control study included 50 epileptic children aged 8-12 years, with a mean age of 9.35 ± 1.59 years and a male to female ratio of 1.8 : 1. They were divided into two subgroups according to the types of seizures: 26 patients with generalized seizures in subgroup I and 24 patients with partial seizures in subgroup II, and 50 apparently healthy children of matched age, sex, and social class were included as the control group. The Arabic version of the 23-item Pediatric quality of life Generic Core Scale (Parents' scale) was applied to evaluate the health-related quality of life of both patients and healthy controls.
Results
Diminished health-related quality of life is a common feature of epilepsy. Highly significant lower overall quality of life scores of all functioning domains of health-related quality of life were present between patients' subgroups (
P
< 0.001), and between total patients versus control groups (
P
< 0.001). Univariate analysis was performed to identify significant predictors of poorer quality of life in children with epilepsy. On analyzing the risk factors using odds ratio, epilepsy-related risk factors such as age at onset of seizures, types of seizures, duration of the illness, number and duration of antiepileptic drugs as well as children's clinical variables such as learning problems, developmental delay in milestones, limitation in child hobbies, urine incontinence, and prolonged sleep, and some family-related variables such as marital disharmony and parental anxiety were found to be significantly strong predictors of poorer health-related quality of life in children with epilepsy, with prediction of 95%.
Conclusion
Epileptic patients were found to be at a higher risk of developing impaired quality of life with lower mean scores of all domains of quality of life, especially patients with generalized, frequent fits, those on polytherapy, and patients with a younger age of seizure onset and a longer duration of illness. The scope of management of epilepsy should include optimal seizure control and improvement of the health-related quality of life of the affected children.
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5
2,920
278
Assessment of compliance to standard precautions among surgeons in Zagazig University Hospitals, Egypt, using the Health Belief Model
Eman M Mortada, Marwa M Zalat
January-June 2014, 9(1):6-14
DOI
:10.4103/1687-4293.137319
Background/aim
Although it is recommended that healthcare professionals should comply with the standard precautions to prevent acquiring blood-borne diseases (AIDS, hepatitis B and C), yet, they frequently do not comply with this recommendation. Understanding the reasons for compliance and noncompliance will help in designing educational programs for hospital staff and in determining a strategy for improving health behavior. The present study aimed to assess surgeons' compliance to standard precautions and determine surgeons' perceived beliefs affecting their compliance using the Health Belief Model.
Participants and methods
A cross-sectional study was carried out at surgical departments in Zagazig University Hospitals from December 2012 to May 2013. A questionnaire on various aspects of infection control and standard precautions practices was provided to 307 surgeons, with a response rate of 70%.
Results
Our findings indicated that 57.5% of the surgeons sampled in Zagazig University Hospitals were compliant with standard precautions. 59.8% of compliant surgeons had been exposed to at least one needle-stick injury in the previous 3 months, whereas slightly less than half of compliant surgeons (48.4%) had been exposed to splashes, with a highly significant difference compared with nonexposed surgeons. All Health Belief Model subscales were correlated directly with the surgeons' compliance, except perceived barriers.
Conclusion
There is adequate compliance with standard precautions among surgeons in Zagazig University Hospitals, especially female surgeons, with a high level of knowledge among compliant compared with noncompliant surgeons. All Health Belief Model subscales were correlated directly with the surgeons' compliance, except perceived barriers. Adequate training of surgeons, provision of infection prevention equipment, regular reporting, follow-up, and assessment of occupational exposures need to be introduced.
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3,917
432
Determinants of unintended pregnancy and its impact on the health of women in some governorates of Upper Egypt
Ammal M Metwally, Rehan M Saleh, Amira M Abdelhamed, Somia I Salama, Carine W Mores, Fatma A Shaaban, Osama M Azmy
January-June 2015, 10(1):1-8
DOI
:10.4103/1687-4293.159368
Background/aim
Worldwide, more than half of all unintended pregnancies end in abortion, indicating the preference for increasingly smaller families. The present work aimed to assess the determinants of unintended pregnancy and its impact on women's health in El Fayoum and Benisuef governorates.
Participants and methods
A community-based cross-sectional household survey was conducted among 827 married women of reproductive age. The study was conducted in two governorates, El Fayoum and Benisuef, for a period of 6 months.
Results
The current study revealed that 15.9% of women who participated in the study had an unmet need for family planning, with subsequent unintended pregnancy. Higher age of women and her age at the time of marriage, illiteracy, short interpregnancy spacing, exceeding the desired number of children, negative attitude of husbands toward the use of family planning methods, and absence of discussion between partners on the use of means to postpone pregnancy increased the number of unintended pregnancies. Also, women with unintended pregnancy were at a higher risk of experiencing health problems during her pregnancy.
Conclusion
Despite the efforts taken by family planners, the problem of unintended pregnancy continues to increase in Egypt, with its adverse effect on pregnancy and maternal outcomes. Therefore, the strategy for maternal mortality reduction in Egypt should focus on addressing the unmet needs of high-parity, uneducated, nonworking women.
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5
2,798
353
Improving the production of unsaturated fatty acid esters and flavonoids from date palm pollen and their effects as anti-breast-cancer and antiviral agents: An
in-vitro
study
Kadry Z Ghanem, Manal M Ramadan, Hassan Z Ghanem, Mohamed Fadel
July-December 2015, 10(2):47-55
DOI
:10.4103/1687-4293.175555
Background/aim
Pollens from different plants contain unsaturated fatty acid esters (USFAEs) and flavonoids that play a very important role as bioactive compounds. Therefore, the present study was designed to improve the production of volatile USFAEs and flavonoids from date palm pollen (DPP) in a culture of
Trichoderma koningii
and test its activities as an anti-breast-cancer and antiviral agent.
Materials and methods
The volatile esters of fermented and nonfermented date palm pollens (FDPPs) were identified using gas chromatographic-mass spectrometric (GC-MS) analysis. Antioxidant activities were determined using three different methods: the 2,2′-diphenyl-1-picrylhydrazyl (DPPH) assay, the ferric reducing antioxidant power assay, and the 2,2-azinobis(3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS) assay. Polyphenols (phenolics and flavonoids) were also determined. Anti-breast-cancer and antiviral activities were determined using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay.
Results
GC-MS analysis showed an improvement in the level of USFAE in FDPP (47.99%) almost double that of the DPP results (24.11%) extract concentration. Flavonoids content of the FDPP extract (93.4 ± 6.3 mg/ml) was higher than that obtained by the DPP extract (45.4 ± 2.1 mg/ml), which was more than double the value. Antioxidant activity of the FDPP extract increased 3.16, 3.42, and 2.14 times that of the DPP extract as determined by the ABTS, ferric reducing antioxidant power (FRAP), and DPPH assays, respectively. The extract of FDPP showed strong anticancer activity against the MCF-7 cell line (IC
50
: 9.52 μg/ml) compared with the DPP extract (IC
50
: 96.22 μg/ml). Also, the FDPP extract had strong antiviral activity (CC
50
: 16.5 μg/ml) compared with DPP (CC
50
: 38.8 μg/ml). This is the first report in which the FDPP extract is used in biological studies as anti-breast-cancer and antiviral agents.
Conclusion
Fermentation of DPP by
T. koningii
improves many bioactive volatile USFAE and flavonoid contents; these have anti-breast-cancer and antiviral activity.
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3
2,759
308
Ameliorative role of ethanolic extract of
Moringa oleifera
leaf on aflatoxin B
1
-induced genotoxicity and biochemical alterations in rats
Ibrahim M Farag, Hanaa M Roshdy, Hasnaa A Radwan, Inas S Ghaly, Sherifa H Salah, Emam A Abdel-Rahim, Aboelfetoh M Abdalla
January-June 2018, 13(1):60-70
DOI
:10.4103/jasmr.jasmr_33_17
Background/aim
The present study was conducted to assess the ameliorative role of
Moringa oleifera
leaf extract (MOLE) on genotoxicity and biochemical alteration of aflatoxin B
1
(AFB
1
) in rats.
Materials and methods
The rat groups involved negative control, control of DMSO, positive control that received AFB
1
in DMSO (0.7 g/kg, body weight) four times weekly for 1 month, groups 4–6 that received the same dose of AFB
1
in DMSO at the same period plus MOLE doses (3.3, 4.0 and 4.7 g/kg) daily for 1 month, and groups 7–9 that received MOLE alone at the same doses for 15 days after cessation of AFB
1
treatment. Molecular genetic, cytogenetic, sperm, and biochemical studies were documented.
Results
Genetic and sperm results revealed that AFB
1
treatment induced significant elevation of genetic alterations and sperm abnormalities as compared with normal control. Biochemical studies showed that the treatment with AFB
1
disturbed the parameters of liver functions, where aspartate-transaminase, alanine-transaminase, and alkaline phosphatase were activated and bilirubin contents as well as the rate of malondialdehyde were increased significantly, but the endogenous antioxidative system (catalase, superoxide-dismutase activities and glutathione as well as total antioxidant capacity) and protein profile were reduced significantly. Moreover, kidney functions (urea, uric acid, and creatinine contents) were elevated under AFB
1
administration. The treatment with MOLE significantly minimizes the genetic alterations, sperm abnormalities, and biochemical destruction. These ameliorations were increased by increasing the dose level. Better findings were seen by using MOLE as a therapeutic agent than its using as a protective agent.
Conclusion
This study revealed that MOLE contains therapeutic factors used in curing of genotoxicity induced by AFB
1
in rats, and treatment of animals that were exposed to AFB
1
with MOLE significantly ameliorated the genetic, sperm, and biochemical parameters as compared with animals treated with AFB
1
alone.
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263
ORIGINAL ARTICLE: BIOCHEMISTRY
Brain-derived neurotrophic factor and coenzyme Q10 levels in blood of children with learning disorder
Mohamed E Elhadidy, Ola H Gebril, Adel Hashish, Ayman Kilany, Neveen H Nashaat, Ehab R Abdelraouf
July-December 2019, 14(2):124-129
DOI
:10.4103/jasmr.jasmr_15_19
Background
Learning disorder (LD) is manifested by significant difficulties in the acquisition and use of reasoning, reading, writing, or mathematical abilities, despite average intelligence and proper education. Its etiological factors were suggested to be related to neurodevelopmental alterations. Measurement of the levels of brain-derived neurotrophic factor (BDNF) and coenzyme Q10 (CoQ10) was targeted in children with LD in comparison with typically developing (TD) ones.
Materials and methods
This study included 82 Egyptian Arabic-speaking children matched for age and sex and socioeconomic status, comprising 42 with specific LD (group I) and 40 TD children (group II). All participants were subjected to clinical and full neurological examination after reporting a full medical history. Furthermore, LD group was subjected to Stanford–Binet intelligence scale, dyslexia assessment test, and phonological awareness test, which evaluates cognitive and learning aptitudes. The levels of BDNF and CoQ10 were determined in serum by enzyme-linked immunosorbent assay.
Results
All children with LD obtained a score of 1 or more as at-risk quotient by the dyslexia assessment test, which indicated a specific reading disorder. The BDNF and CoQ10 levels in the LD group were significantly less than those in the TD group. No correlations were found between the measured markers and each other or between them and the measured factors of the used tests.
Conclusion
The detected low levels of BDNF and CoQ10 in children with specific LD with impairment in reading would be suspected to be related to etiological or exaggerating factors for the deficits in such children.
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1,180
109
ORIGINAL ARTICLES
VELscope versus toluidine blue for detection of dysplastic changes in oral keratotic lesions: diagnostic accuracy study
Mostafa Belal, Wesam A Elmoneim, Sherine Nasry, Basma Mostafa, Shereen Ali
January-June 2018, 13(1):45-52
DOI
:10.4103/jasmr.jasmr_2_18
Background/aim
Various techniques have been implemented in the early detection of oral potentially malignant disorders including commonly occurring hyperkeratotic lesions. The objective of the present study was to assess the efficacy of visually enhanced lesion scope (VELscope Vx) versus toluidine blue (TB) with reference to histopathological examination in the detection of dysplastic changes in oral keratotic lesions.
Patients and methods
A total of 30 patients having oral keratotic lesions were clinically diagnosed and subjected to VELscope Vx and TB examination. Histopathological assessment of biopsied lesions was also done. Sensitivity, specificity, positive and negative predictive values were obtained for both VELscope and TB and were compared with histopathological examination.
Results
VELscope Vx was able to identify four out of seven cases with mild dysplasia and a case of oral squamous cell carcinoma, while TB was able to identify five cases and the oral squamous cell carcinoma case as true positive cases. Thus, sensitivity was 62.5 and 75% for VELscope Vx and TB, respectively. Regarding the specificity of VELscope Vx, it was 71.4% while TB had a specificity of 85.7%.
Conclusion
The sensitivity and specificity of TB examination outweighed those of VELscope Vx in detecting dysplastic changes within keratotic lesions.
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2
2,500
271
The antifibrotic effect of Zilla spinosa extracts targeting apoptosis in CCl4-induced liver damage in rats
Enayat A Omara, Sayed A El-Toumy, Marwa E Shabana, Abdel-Razik H Farrag, Somaia A Nada, Nermeen Shafee
July-December 2018, 13(2):129-143
DOI
:10.4103/jasmr.jasmr_29_18
Background/aim
Liver fibrosis and its end-stage cirrhosis are the main reasons of morbidity and mortality all over the world. The current study aimed to evaluate the efficacy of
Zilla spinosa
(
Z. spinosa
) on CCl
4
-induced liver fibrosis, apoptosis, and oxidative stresses in rats.
Materials and methods
Extract of aerial part of
Z. spinosa
was used in this study. Thirty male Sprague‑Dawley rats were enrolled in this study and divided into five groups (six each): group 1 served as control and groups 2–5 were treated with CCl
4
(1 ml/kg intraperitoneal twice a week for 8 weeks), where group 2 served as a control positive, group 3 received silymarin (50 mg/kg) daily, and groups 4 and 5 were administrated with
Z. spinosa
(100 and 200 mg/kg, respectively) daily for 8 weeks. At the end of each experiment, liver function tests were analyzed in serum, whereas malondialdehyde (MDA), Nitric oxide (NO), Glutathione (GSH), and hydroxyproline (HA) were analyzed in liver tissues. Liver fibrosis was confirmed histopathologically, and collagen content, caspase-3, and α-smooth muscle actin (α-SMA) were assayed immunhistochemically.
Results
Alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, total bilirubin, MDA, NO, and HA levels were increased (
P
<0.05), whereas total protein and GSH were decreased (
P
<0.05) in CCl
4
-administrated rats. Histopathological results showed loss of lobular structure, fibrosis with expansion of portal tract by fibrous tissue together with inflammatory changes confined to portal tract and central vein, and intense centrilobular necrosis and remarkable fatty hydropic degeneration. In addition, extensive accumulation of connective tissue, marked depletion of glycogen, strong expression of α-SMA, and increased of caspase-3 were found in CCl
4
-administrated rats. Oral administration of
Z. spinosa
at 100 or 200 mg/kg restored the normal levels of liver function parameters, MDA, NO and GSH; decreased HA; and reduced collagen, glycogen content, caspase-3, and α-SMA in liver tissue of rats. The high dose of 200 mg/kg showed more potent effect than low dose of 100 mg/kg when compared with silymarin treatment group.
Conclusion
The present study clarified that
Z. spinosa
extract has antioxidant and antiapoptotic properties in CCl
4
-induced liver fibrosis in rats, and may be able to exert a therapeutic effect on developing hepatic fibrosis; moreover, high dose of 200 mg/kg appeared to be more potent than low dose (100 mg/kg).
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2
1,744
177
Role of citicoline as a protective agent on toluene-induced toxicity in rats
Nermeen Shaffie, Marwa E Shabana
January-June 2019, 14(1):14-24
DOI
:10.4103/jasmr.jasmr_9_19
Background/aim
Toluene is used as an organic solvent; it has toxic effects on liver, renal, and neurological tissues. Citicoline has antioxidant effects. The aim of this study was to investigate the protective effect of citicoline against toluene-induced toxicity in rats.
Material and methods
A total of 24 rats were divided in four equal groups with six rats each. Rats in group 1 were the control. Rats in group 2 were exposed to toluene intraperitoneally at 900 mg/kg. Rats in group 3 were exposed to toluene 900 mg/kg in combination with citicoline at 150 mg/kg orally. The rats in group 4 were exposed to toluene 900 mg/kg in combination with citicoline 300 mg/kg orally. All treatments were used daily for 6 days. All rats were killed by decapitation. Tissue sections were stained with routine histological methods and examined under light microscope. In addition, sections were immunohistochemically staining with caspase-3 for liver and renal tissues and glial fibrillary acidic protein (GFAP) for brain tissue, and area percentage of positivity was measure by image analysis system.
Results
Histopathological and immunomorphometric studies of GFAP in brain and caspase-3 in liver and kidney were done. Toluene-treated groups showed vacuolar degeneration of hepatocytes and epithelial lining of renal tubules, neuronal damage, and neurodegeneration. There was an increase in caspase-3 in liver and kidney, which show marked increase in control positive rats that treated with toluene alone, but when large dose of citicoline 300 mg was added to toluene, the area percentage of caspase-3 was markedly decreased in liver and kidney. In addition, GFAP expression in brain tissue was decreased in toluene-treated control positive group, which then increased when treated with citicoline especially with a large dose.
Conclusion
The results of this study indicate that citicoline treatment can protect against toluene-induced toxicity in rats.
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Cytotoxicity and antimicrobial activity of naturally and chemically synthesized zinc oxide nanoparticles
Amr A El-Waseif
January-June 2019, 14(1):42-51
DOI
:10.4103/jasmr.jasmr_8_19
Background/aim
Zinc oxide (ZnO) is a polar inorganic compound with numerous applications, for example, as an antimicrobial agent. The present study aims to synthesize ZnO nanoparticles (NPs) by two different methods, and then determine the antimicrobial activity of ZnO NPs from both methods. It then focuses on comparison between the cytotoxicity of both ZnO NPs.
Materials and methods
ZnO NPs were synthesized using natural and chemical methods. The synthesized and prepared ZnO NPs were detected by precipitation in both methods using de Man, Rogosa, and Sharpe broth and alkaline medium, respectively. The characterization of ZnO NPs was performed using ultraviolet spectroscopy, zeta potential, and transmission electron microscopy (TEM) to decide properties of NPs. Viability tests are essential for assessing the effect of toxicants on cells. To measure cell viability following NP exposure, MTT assay was used.
Results
Results of ultraviolet and TEM experiments for both NPs indicated absorbance at 356–360 nm, which is typical for ZnO NPs. Results show that naturally prepared ZnO NPs had an average size within 7.8 nm; they were small spherical particles with a narrow size distribution relatively with smooth surfaces. The chemically prepared ZnO NPs’ TEM images showed an average size of 27.6 nm. Zeta value of naturally synthesized ZnO NPs was estimated to be −25.30 mV at pH=7. However, the value of zeta potential in chemical preparation strategy showed −18.6 mV. Results revealed that toxicity of naturally synthesized ZnO NPs was less than that of chemically prepared ZnO NPs. Furthermore, the decline in cytotoxicity attributed to ZnO NP exposure was dependent on the concentration of ZnO NPs. The antimicrobial activity results of ZnO NPs showed that the ZnO NPs produced from both methods recorded antimicrobial activities against the pathogenic strain models used.
Conclusion
The microbial synthesized ZnO NPs within size 7.8 nm when used at concentration 625 μg/ml as antimicrobial agent recorded the lowest cytotoxicity when compared with chemically synthesized. So that natural synthesis of ZnO was recommended.
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1,622
206
Histopathological and histochemical effects of nicotine on the liver and kidney of adult male rats
Medhat M Menshawy, Walid M Sharaf, Reham S.E. Esmail, Abdel R.H. Farrag
January-June 2019, 14(1):7-13
DOI
:10.4103/jasmr.jasmr_39_18
Background/aim
Nicotine is the more abundant alkaloid component contained in tobacco. Smoking causes numerous diseases that are associated with anemia. Nicotine is metabolized primarily by the liver, and to a lesser extent, by the kidney. The aim of this study was to investigate the iron distribution and histopathological changes in liver and kidney tissues of rats administered with nicotine.
Materials and methods
This study was performed on thirty adult male albino rats. Animals were divided into three groups (10 each). Group I served as control rats, which were injected subcutaneously with 1 ml of normal saline; group II rats were injected subcutaneously with nicotine at a dose of 0.25 mg/kg/day; and group III rats were injected subcutaneously with nicotine at a dose of 0.5 mg/kg/day for 8 weeks. At the end of the experiment, the animals were killed under ether anesthesia. Liver and kidney tissues specimens were prepared by routine histological procedures and examined under a light microscope.
Results
Injection with nicotine showed iron deposition inside the liver sinusoids, seen as green bluish stain. In addition, scattered iron intracellular deposition appeared as scattered bluish dots in kidney. Histopathological examination of liver specimens from rats injected subcutaneously with nicotine showed mononuclear cell infiltration and that some of the hepatocytes had a hyperchromatic nucleus and enlarged sinusoids as compared with the control ones. However, kidney specimens showed some vacuolated tubular cells, few dilated tubules, and the presence of intraluminal casts in some tubules.
Conclusion
This study indicated that subcutaneous injection of nicotine causes iron deposition in the liver and kidney and is associated with structural damage.
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249
Serum levels of monocyte chemoattractant protein-1, interlukin-6, and paraoxonase-1 in childhood obesity
Enas R Abd El Hamid, Azza A Abdel-Shaheed, Rania N Sabry, Eman R Youness, Mones M Abu Shady, Amr Said Megawer, Nadia A Mohamed
January-June 2015, 10(1):27-31
DOI
:10.4103/1687-4293.159371
Background/aim
Obesity is becoming an epidemic health problem. Elevated cytokines and chemokines are prominent features in obesity, which play a main role in the development of other chronic diseases. The aim of this study was to determine the serum levels of monocyte chemoattractant protein-1 (MCP-1), interlukin-6 (IL-6), and serum paraoxonase-1 (PON1) in childhood obesity.
Materials and methods
The present study included 40 obese school-aged children (5-15 years) and 40 healthy children as controls. The patients were presented to the outpatient clinic in National Institute of Nutrition. MCP-1, IL-6, PON1, total cholesterol, and triglycerides were measured in all participants.
Results
The mean serum levels of MCP-1, IL-6, and total cholesterol were significantly higher in obese participants than in controls (
P
< 0.0001), whereas the PON1 was significantly lower in obese participants than in controls (
P
< 0.0001). MCP-1, IL-6, and serum cholesterol levels showed significant positive correlation with BMI (
P
< 0.05), whereas PON1 showed a significant negative correlation with BMI (
P
< 0.05). Multiple regression analysis showed a strong association between PON1 activity and BMI (
P <
0.0001).
Conclusion
Childhood obesity is associated with increased serum MCP-1 and IL-6 and decreased PON1 and hypercholesterolemia suggesting an increase in adulthood disease risk. Measuring serum MCP-1, IL-6, PON1 activity in obese children may be a good predictor for future chronic disease development and complications.
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Effect of
Escherichia coli
,
Salmonella
spp., and
Staphylococcus aureus
isolated from polluted drainage water on rats
Gamal A Osman, Medhat M Menshawy, Abdel Razik H Farrag
January-June 2016, 11(1):1-8
DOI
:10.4103/1687-4293.186778
Background/aim
Water pollution not only damages the environment but also kills wildlife. The present study aimed to investigate the effect of
Escherichia coli
,
Salmonella typhi
, and
Staphylococcus aureus
, which were isolated from drainage water at El-Rahawey area, Giza, Egypt, on experimental rats.
Materials and methods
Water samples were collected from the subsurface layer for bacteriological examination, stored in an ice box, and delivered to the laboratory and analyzed to count the total viable bacterial counts/ml using the most probable number technique. The isolated bacteria were given to rats for 21 days at a dose equal to 10
3
/ml and the liver and kidney were dissected for histopathological and histochcemical studies.
Results
Results showed that the average counts at 22 and 37°C were 10
5
and 10
6
CFU/ml, respectively, whereas the average counts by using the most probable number technique/100 ml were 10
5
, 10
4
, 10
3
, 10
3
, and 10
2
for total coliforms, fecal coliforms,
E. coli
,
Salmonellae
group, and total staphylococci, respectively. Isolation of pathogens from water sources were identified by using membrane filter technique on specific media. The histopathological examination of the liver of treated rats with
E. coli
,
S. typhi
, and
S. aureus
revealed swollen hepatocytes with decreased sinusoidal spaces and widely distributed necrotic foci. In the kidney, renal tubules showed extensive epithelial swelling with decreased lumen space and generalized necrotic changes with interstitial hemorrhage in renal cortex. The histochemical study indicated the depletion of staining of protein and polysaccharides.
Conclusion
E. coli
,
S. typhi
, and
S. aureus
that are isolated from polluted drainage water cause histological and histochemical changes in the liver and kidney of rats.
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282
Ultrastructural and morphometric effect of aluminum phosphate and calcium phosphate nanoparticles as adjuvants in vaccinated mice
Mohamed S Salim, Aliaa M Issa, Abdel Razik H Farrag, Hamdalla H Zidan, Aly F Mohamed
January-June 2016, 11(1):22-28
DOI
:10.4103/1687-4293.186780
Background/aim
Calcium phosphate (CAP) and aluminum phosphate (alum) compounds have been approved as vaccine adjuvants for humans. The present study aimed to study the ultrastructural and morphometric effect of both aluminum phosphate (alum) and CAP nanoparticle adjuvant.
Materials and methods
Eighteen Swiss albino mice were used in the experiment. An overall 50% of them were adult and the other 50% were juvenile. Mice were immunized intramuscularly with 0.125 ml of adjuvanted tetanus toxoid vaccine. In the alum adjuvant group, three adult mice and another three juvenile ones were injected with alum adjuvanted vaccine and killed 1 week after immunization. In the CAP adjuvant group, three adult mice and another three juvenile mice were injected with CAP adjuvanted vaccine and killed 1 week after immunization. At the end of the work, samples of the liver, kidney, and brain were subjected to light and electron microscopic examinations.
Results
In both the alum group and the CAP nanoparticle adjuvant group, the examination revealed wide and severe ultrastructural changes in the liver, kidney, and brain of adult and juvenile mice. These changes included swollen mitochondria with degenerated cristae, severely degenerated ground cytoplasm, and unusual chromatin picture of nucleus. In contrast, morphometric studies of the nucleus/cell ratio and damaged areas showed a significant increase in the liver, kidney, and brain of both adjuvants.
Conclusion
The results of this study revealed that both aluminum phosphate (alum) and CAP nanoparticle adjuvant caused ultrastructural and morphometric changes in tissues of the liver, kidney, and brain.
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Differential effects of t helper-17 cytokines on the functions of granulocytes isolated from
schistosoma mansoni
-infected patients and healthy individuals
Ayatollah El-Shorbagy, MT Shata, Mohammad A Mohey, Soad Nady
January-June 2016, 11(1):29-36
DOI
:10.4103/1687-4293.186781
Background and aim
The role of T helper-17 lymphocytes in the regulation of the immune response against
Schistosoma
soluble egg antigens is still controversial. In this study, the in-vitro effects of T helper-17 cytokines [interleukin (IL)-17 and IL-22] on granulocyte functions isolated from
Schistosoma-
infected patients or healthy individuals were examined.
Patients and methods
Twenty-seven
Schistosoma
mansoni-
infected patients and 13 healthy individuals from Kasr Al-Aini Viral Hepatitis Center were enrolled in the present study. Granulocytes were isolated from whole blood of patients and controls by Ficoll–Paque density gradient for removal of the mononuclear layer and then lysis of red blood cells. Granulocytes were stimulated
in vitro
with soluble egg antigen in the presence of IL-17, IL-22, or both. After 24 h, the supernatants were collected for the measurement of tumor necrosis factor (TNF)-α, hydrogen peroxide (H
2
O
2
), myeloperoxidase (MPO), and nitric oxide (NO) using enzyme-linked immunosorbent assay as surrogate markers for granulocyte functions.
Results
The results indicated that the presence of IL-17 significantly decreased (
P
<0.05) TNF-α, H
2
O
2
, MPO, and NO production by granulocytes isolated from
Schistosoma-
infected patients. In contrast, in the presence of IL-22 or both IL-17 and IL-22, there were significant increases in the production of H
2
O
2
and TNF-α by granulocytes isolated from
Schistosoma-
infected patients. Moreover, in the presence of both IL-17 and IL-22, nonsignificant changes were observed in MPO or NO levels compared with those in the control participants.
Conclusion
IL-17, in contrast to IL-22, inhibited the functional activity of granulocytes isolated from
S. mansoni-
infected patients. Therefore, neutralization of IL-17 may work as a therapeutic strategy for these patients.
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1,529
160
Studying of B-cell differentiation and macrophage colony-stimulating factors as new players in the diagnosis of rheumatoid arthritis
Mohamed A Mokhles, Hala Z Raslan, Hiba Sibaii, Salwa R El-Zayat, Mahitab El-Kassaby, Hagar Hassan
July-December 2017, 12(2):79-85
DOI
:10.4103/jasmr.jasmr_21_17
Background/aim
There is an imperative, yet unmet need for the development of a biomarker panel for progression of rheumatoid arthritis (RA). This study aimed to investigate the reliability of B-cell differentiation factor (BCDF) and macrophage colony-stimulating factor (MCSF) as diagnostic and prognostic biomarkers of RA.
Patients and methods
A total of 25 RA patients were enrolled and divided into three subgroups according to the disease activity score, remission/minimal, moderate, and severe, in addition to 20 healthy controls. Enzyme-linked immunosorbent assay was used to measure serum BCDF, MCSF, and interleukin (IL)-2, and the calorimetric method was used to measure immunoglobulin G (IgG). In addition, the erythrocyte sedimentation rate was measured at 1 h in the RA patients.
Results
BCDF, MCSF, and IgG were significantly higher in the RA patients compared with the controls. BCDF was significantly higher in the severe disease group compared with the remission/mild and the moderate disease groups, whereas IgG was significantly higher among the moderate disease group compared with the remission/mild disease group, and among the severe disease group compared with the remission/mild disease group, whereas the erythrocyte sedimentation rate did not show any significant difference between the three subgroups of the disease group. BCDF delineates RA from controls at a cut-off value of 102 pg/ml, whereas at a cut-off value of 37.5 ng/l MCSF delineates RA from controls.
Conclusion
Serum BCDF and MCSF can be used as RA diagnostic biomarkers, whereas BCDF can be used additionally as a prognostic biomarker.
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176
Role of a combination of myrtle extract and fish oil supplementation in improving endothelial dysfunction in streptozotocin-induced diabetic rats
Omnia Aly, Tahany R Elias, Mervet H Agaibyi, Wafaa I Rasheed, Magdi N Ashour, Jihan S Hussein
July-December 2017, 12(2):86-91
DOI
:10.4103/jasmr.jasmr_26_17
Background/aim
Diabetes mellitus is one of the most important systemic diseases in the world and considered as the leading cause of endothelial dysfunction. Our previous studies have been clarified the antioxidant, anti-inflammatory, and antidiabetic effects of myrtle extract and fish oil. This study aimed to explore the effect of supplementation of a combination of myrtle extract and fish oil in improving the endothelial dysfunction associated with diabetes.
Materials and methods
Sixty albino rats were classified into three groups (20 rats each): control group, diabetic group, and treated diabetic group that received myrtle extract and fish oil. Plasma 5-lipoxygenase (5-LOX), interleukin-1β, tumor necrosis factor α, lipoxin-A4, asymmetric dimethylarginine, nitric oxide, glucose, insulin, insulin resistance, and cell membrane fatty acids [arachidonic acid (ω-6), linoleic acid (ω-6), α-linolenic acid (ω-3), and oleic acid (ω-9)] were estimated.
Results
Using a combination of myrtle extract and fish oil induced significant decrease in the plasma level of 5-lipoxygenase, asymmetric dimethylarginine, arachidonic acid (ω-6), linoleic acid (ω-60), and glucose, whereas induced significant increase in lipoxin-A4, nitric oxide, and α-linolenic acid (ω-3).
Conclusion
A combination of myrtle extract and fish oil has a potential effect that exceed the beneficial role of each of them in reducing endothelial dysfunction as well as oxidative stress and inflammation associated with diabetes mellitus.
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Behavioral determinants of hand hygiene compliance among physicians in Riyadh region in Saudi Arabia
Hala A Amer, Abdalla F Bahar, Muhammad N Algamal, Hind A Alzoman, Ammal M Metwally
July-December 2017, 12(2):99-105
DOI
:10.4103/jasmr.jasmr_23_17
Background/aims
Hand hygiene is recognized globally as a leading measure to reduce healthcare-associated infection. However, compliance with hand hygiene is unacceptably low. Physicians − in particular − have repeatedly been observed to be poor compliers. Promotion of this important practice is a complex issue. It is important to study the behavioral determinants in relation to hand hygiene practices to develop effective improvement strategies. The aims of our study are to assess the knowledge of the physician and their attitudes and practices of hand hygiene, to identify the behavioral determinants of noncompliance, and to recommend interventions that could increase the practice.
Participants and methods
An anonymous questionnaire was administered that included four sections: participants’ demographic data, self-reported compliance on the basis of the Centers for Disease Control and Prevention Guidelines, attitudes in relation to the studied behavior adapted from a validated published questionnaire, and the knowledge assessment section chosen from the WHO questionnaire. The questionnaire was distributed by infection control practitioners at King Saud Medical City − a tertiary care hospital in Riyadh, Saudi Arabia − and self-completed by 127 physicians of different specialties during March–May 2015.
Results
The response rate was 75%. The mean self-reported compliance rate was 89.4%. A multiple linear regression analysis was carried out on the studied variables that identified three main predictors most likely to be correlated with hand hygiene behavior noncompliance: self-efficacy (
β
=0.252;
P
=0.004), time-related attitude (
β
=0.16), and usefulness-related attitude (
β
=0.148).
Conclusion
Understanding the determinants of a fundamental behavior such as hand hygiene in healthcare settings is among the essential steps to plan more effectual intervention strategies for compliance improvement. Contributions from behavioral and social sciences are crucial when designing studies to investigate and improve behavior acquiescence. The healthcare system is complex and more studies are needed to elaborate specific behavioral determinants in relation to the practice of hand hygiene in other variable work situations and among different healthcare workers categories and employee belong to different culture group.
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150
Carotid intima–media thickness assessment in obese patients with chronic renal failure
Sahar A El-Masry, Hosneya A Mohamed, Ashraf M Enaite, Manal M Ali, Ibrahim A Ibrahim
January-June 2018, 13(1):18-24
DOI
:10.4103/jasmr.jasmr_16_18
Background/Aim
Measurement of carotid artery intima–media thickness (CIMT) is reliable for early detection of atherosclerosis, one of obesity’s complications, which is a leading cause of mortality among patients with end-stage renal disease. The study aimed to evaluate CIMT in relation to obesity, chronic renal failure (CRF) and both, for the early prevention of cardiovascular problems.
Patients and methods
This cross-sectional study included 118 adult individuals of both sexes, aged 30–60 years. Patients with CRF were gathered from the renal dialysis unit of King Fahd Hospital-Kasr El Einy Hospital, and the obese without CRF from the ‘Management of Visceral obesity Unit’, in ‘Medical Excellence Research Center (MERC)’, National Research Centre, during the period spanning from June 2015 to April 2016.They were grouped into three groups: a case group, which included 45 obese individuals with CRF, and two control groups: one comprised 39 nonobese patients with CRF and the other group comprised 34 obese patients without CRF. Anthropometric assessment, lipid profile, and ultrasound measurement of CIMT were performed for each patient.
Results
CIMT was greater in all groups than the normal range (0.06–0.08 cm), particularly in the obese group without renal failure. CIMT had a highly significant correlation with waist circumference and insignificant correlation with BMI and lipid profile in different groups.
Conclusion
The increase in CIMT was related to obesity and renal failure, but it was more prominent with obesity. CIMT had highly significant correlation with central obesity in cases wherein obesity and renal failure were coexisting together and insignificant correlation with lipid profile.
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1,732
195
Autologus serum and sodium hyaluronate role in alkali corneal burn healing
Eman S Mohamed, Mervat A Ali
January-June 2018, 13(1):39-44
DOI
:10.4103/jasmr.jasmr_13_18
Background/aim
Corneal burn wound healing includes a sequence of difficult processes that are focused on improving the outcomes, in particular, the healing time and the feature of the scar. Bodily fluids such as natural tears and autologus serum were used in the treatment of corneal burn. The aim of the study was to investigate the cure of corneal burn alkali injury by autologous serum (AS) and/or sodium hyaluronate.
Materials and methods
A total of 50 Wister rats (200–250 g) were distributed into five groups: group I acted as the control, group II had alkali burn (AB) with sodium hydroxide, group III had AB treated with AS, group IV was AB treated with sodium hyaluronate, and group V had AB with sodium hydroxide treated with autologus serum and sodium hyaluronate. Comet assay analysis was applied for cornea to determine DNA damage. Malondialdehyde, superoxide dismutase, glutathione peroxidase, and catalase were measured to estimate the balance between oxidants and antioxidants in the cornea.
Results
The data indicated a significant increase (
P<
0.05) in all comet assay parameters and malondialdehyde level in addition to a significant decrease (
P<
0.05) in superoxide dismutase, glutathione peroxidase, and catalase activity owing to AB. Enhancements of measured parameters were observed in all other treated groups.
Conclusion
AS accelerates the AB healing process, but the process was faster when diluted by sodium hyaluronate.
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1,787
144
REVIEW ARTICLE
Development of immunoassays of catecholamines and their metabolites
Masanori Yoshioka
July-December 2016, 11(2):37-42
DOI
:10.4103/1687-4293.199296
Catecholamines take the role of neurotransmitters and hormones and are involved in many diseases. Sensitive immunoassays for catecholamines and their metabolites were required to study the diagnoses of diseases. First, we achieved success in producing specific antibodies for catecholamines and basic metabolites using each antigen. We applied a monoclonal antibody to 3-methoxy-4-hydroxyphenylglycol to measure the concentrations to diagnose depression. The acidic homovanillic acid (HVA) and d-3-methoxy-4-hydroxymandelic acid (VMA) were useful in mass screening of neuroblastoma using monoclonal antibodies. The paper reviews these developments for future use. An antigen is important to produce the specific antibody. The amino acid residue of each catecholamine and l-3,4-dihydroxyphenylalanine protected with
N-
maleyl group as well as basic metabolites was reacted with bovine serum albumin using Mannich reaction in the presence of formaldehyde. The
N-
maleyl group of the conjugate was moderately liberated to give rise to the antigen. The antigen was injected with Freund’s Complete Adjuvant to the rabbit or mouse. The conventional or monoclonal antibody was used for radioimmunoassay or enzyme immunoassay. Each immunoassay showed high specificity in discriminating not only the fine structure of the hapten but also body ingredients. The kits of HVA and VMA were useful in screening the urine from the infant with neuroblastoma. The secret of specific antibody preparation is dependent on the synthetic method of the conjugation, which is chemically moderate. The preparation of antibody requires long time to increase the affinity. Thus, the methods of antibody preparation are established and repeatable for anyone. The mass screening of neuroblastoma with HVA and VMA was easily applied to infants. Our previous results had established immunoassays for all members of catecholamines and their metabolites for diagnosis and research.
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