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Standarizing in Planta Agrobacterium Tumefacience Mediated Genetic Transformation Protocol to Develop New Events by Transforming G. Hirsutum Cotton based on Cry1Ac-Cry1Ec Genes
Yanal A. Alkuddsi,
Shreekanth S. Patil,
S. M. Manjula,
K. J. Pranesh,
B. C. Patil
Issue:
Volume 2, Issue 4, August 2014
Pages:
190-199
Received:
13 July 2014
Accepted:
22 July 2014
Published:
10 August 2014
Abstract: Cotton breeding for insect resistance has been limited by a lack of sufficient genetic variation in the existing germplasms. Therefore, genetic engineering provides the possibility of creating varieties carrying new properties coming even from heterologous source. Exogenous pesticidal transgenes can be introduced into plants. Agrobacterium mediated plant transformation offers advantages like reducing copy number of the transgene and little co-suppression. Inter specific hybrids are known to be more susceptible to biotic stress. It is hence important to develop Bt version for inter specific hybrid. Presently, the Bt gene commercialized are owned by private sector. It is necessary to develop public sector’s Bt event and commercialize them. UAS Dharwad is involved in developing public sector Bt cotton genotypes. One variety, RCR4 (Gossypium hirsutum, L.) was used in the present investigation. Cry1Ac-Cry1Ec genes are to control Helicoverpa armigera and Spodoptera litura. The seedlings in pots were co-cultivated with solid Agrobacterium culture after cutting the meristematic tip with sharp knife. The number of seedlings co-cultivated, number of seedlings established and the number of seedlings showing transformed status are presented in this study. PCR was performed to confirm the presence of the transgene in the plants that were selected to be advanced further. The results showed that non of plants had trangenes Cry1Ac- Cry1Ec as detected through PCR amplification. In planta genetic transformation was carried out and the plants were tested in T0 generation by means of PCR amplification for the genes Cry1Ac- Cry1Ec. The results obtained were not amplified the Cry1Ac- Cry1Ec. Hence the transformation of the genes was not up to mark and the plants of T1 generation are also not confirmed.
Abstract: Cotton breeding for insect resistance has been limited by a lack of sufficient genetic variation in the existing germplasms. Therefore, genetic engineering provides the possibility of creating varieties carrying new properties coming even from heterologous source. Exogenous pesticidal transgenes can be introduced into plants. Agrobacterium mediated...
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Asymptomatic Carriage of Plasmid-Mediated Multidrug-Resistant Staphylococcus aureus in Nasal Tracts of Persons in a Semi-Urban Area of Nigeria
Jonathan Osariemen Isibor,
Ehima Otabor
Issue:
Volume 2, Issue 4, August 2014
Pages:
200-204
Received:
22 July 2014
Accepted:
30 July 2014
Published:
10 August 2014
Abstract: Staphylococcus aureus remains one of the most important opportunistic bacterial pathogens of man and strains resistant to available drug classes have become more and more prevalent in clinical and community settings. The aim of this study was to determine the nasal carriage rates of multidrug – resistant (MDR) Staph aureus in food vendors and hospital workers in Ekpoma, Nigeria, and to assess whether resistance was plasmid – mediated. With the use of sterile swab sticks (Sterilin, UK), one hundred nasal swabs were obtained from consenting volunteers. Each swab was streaked on mannitol salt agar (Difco, Detroit MI) and incubated at 370C for 24hrs. Colonies growing on medium were Gram – stained and tested for catalase, coagulase and mannitol fermentation. Antibiotic susceptibility tests were performed using the Kirby-Bauer disc diffusion technique, and results were interpreted using the Clinical and Laboratory Standard Institute’s guidelines. Results showed that 32 (32%) of the 100 specimens screened yielded Staph aureus, with 18 (36%) and 14 (28%) of the isolates recovered from hospital workers and food vendors, respectively. The prevalence rate in males (40%) compared with females (24%) was not statistically significant (P>0.05), using Student’s t-test analysis. All the Staph aureus isolates were susceptible to Augmentin. The highest resistance rates of 65.6%, 59.3% and 50.0% were recorded for Amoxycillin, Ofloxacin and Oxacillin respectively, while low resistance rates were recorded for Gentamicin (9.38%) and Cefuroxime (15.6%). Of the 17 (53%) MDR isolates of Staph aureus, only 4 (23.5%) remained MDR after curing with 0.1mg/ml acridine orange solution. Observance of personal hygiene can reduce the level of contamination or infection of carriers thereby reducing the risk of infection for others in the community, while rational use of antibiotics by health professionals and end users could help to curb the rising trend of antibiotic resistance within the community.
Abstract: Staphylococcus aureus remains one of the most important opportunistic bacterial pathogens of man and strains resistant to available drug classes have become more and more prevalent in clinical and community settings. The aim of this study was to determine the nasal carriage rates of multidrug – resistant (MDR) Staph aureus in food vendors and hospi...
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Protective Effect of Aspirin on γ Radiation-Induced Sperm Malformations in Swiss Albino Male Mice
Marowa Mansour AlMathkour,
Entissar Suliman AlSuhaibani
Issue:
Volume 2, Issue 4, August 2014
Pages:
205-210
Received:
4 August 2014
Accepted:
11 August 2014
Published:
20 August 2014
Abstract: This study evaluated the effects of γ radiation on spermatozoa based on the morphological characteristics of sperm in the caudal epididymis of SWR/J mice. In this study, various abnormal sperm shapes (amorphous heads, hook-less, mid-piece defect, pinheads, coiled tails, and tail-less) were observed after exposure to γirradiation (2 or 4 Gy)and after treatment with aspirin (ASA) and γ-irradiation (0.5 mg/kg + 2 or 4 Gy, 5 mg/kg + 2 or 4 Gy and 50 mg/kg + 2 or 4 Gy). The higher rate of abnormal sperm forms was observed in the γ-irradiated mice compared with the aspirin + γ-irradiation-treated mice. In addition, the numberof sperm with amorphous heads and coiled tails wassignificantly increasedafter irradiation. This study suggests that ASA can effectively reducethe effects of 2to4-Gy radiationin sperm. However, further studies are needed elucidate the mechanisms underlying the antioxidant effect of ASA.
Abstract: This study evaluated the effects of γ radiation on spermatozoa based on the morphological characteristics of sperm in the caudal epididymis of SWR/J mice. In this study, various abnormal sperm shapes (amorphous heads, hook-less, mid-piece defect, pinheads, coiled tails, and tail-less) were observed after exposure to γirradiation (2 or 4 Gy)and afte...
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Histopathological Alterations in the Gill, Liver and Brain of Cyprinus Carpio on Exposure to Quinalphos
Ramesh Raju Chamarthi,
Manjunatha Bangeppagari,
Jaffer Mohiddin Gooty,
Srinivasulu Mandala,
Juan Ortiz Tirado,
Shambanagouda R. Marigoudar
Issue:
Volume 2, Issue 4, August 2014
Pages:
211-216
Received:
18 March 2014
Accepted:
20 May 2014
Published:
30 August 2014
Abstract: Study was conducted to assess the histopathological damage of Gill, Liver and Brain in common carp, Cyprinus carpio after sublethal exposure to Quinalphos. Exposed to sublethal concentration (One tenth (1/10th, 0.75 μl/L) of commercial grade quinalphos (25% Emulsified Concentration) for 1, 7, 14, 21 and 30 days and a parallel control was run simultaneously. Gill, Liver and Brain of exposed individuals exhibited some remarkable changes in their histology in comparison to control. Prominent changes include shrinkage of the glomerulus, and dilation of tubular lumen. Vacuolization, desquamation, hydropic swelling and hyaline degeneration of tubular epithelium is also observed. Cyst formation and hemorrhage also appear in certain specimens. Duration of exposure appears to have a profound effect on Gill, Liver and Brain as with increasing duration of exposure histopathological damages become more severe.
Abstract: Study was conducted to assess the histopathological damage of Gill, Liver and Brain in common carp, Cyprinus carpio after sublethal exposure to Quinalphos. Exposed to sublethal concentration (One tenth (1/10th, 0.75 μl/L) of commercial grade quinalphos (25% Emulsified Concentration) for 1, 7, 14, 21 and 30 days and a parallel control was run simult...
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Expression of O6 Methyl Guanine Methyl Transferase (Mgmt) in Oral Squamous Cell Carcinoma Patients in Alexandria, Egypt (Smokers and Non-Smokers)
Hend Mohamed Helmy,
Taissir Ali Omar,
Sahar Mohamed ElSheikh,
Ahmed Serag Habib
Issue:
Volume 2, Issue 4, August 2014
Pages:
217-223
Received:
6 August 2014
Accepted:
16 August 2014
Published:
30 August 2014
Abstract: Objectives: O6-methylguanine-DNA methyltransferase (MGMT) removes mutagenic, carcinogenic, and cytotoxic adducts from O6-methylguanine in DNA through a direct reversal mechanism. Decreased expression of MGMT has been reported in a variety of human malignant tumors. The purpose of this study was to clarify the correlation of MGMT expression levels in oral squamous cell carcinoma (OSCC) and cigarette smoking. Study Design: MGMT protein expression in 22 cases of oral squamous cell carcinoma by immunohistochemistry was investigated. Correlation with detailed tobacco history was then tested by statistical analysis. Results: All the non smoker OSCC cases showed positive immunoexpression. However 3 cases of the smoker patient revealed a weak positive immunostaining with only sporadic cells at the periphery. Conclusion: The results suggest that the absence of MGMT expression is related strongly to tobacco smoking and, thus, might be a significant event in oral carcinogenesis.
Abstract: Objectives: O6-methylguanine-DNA methyltransferase (MGMT) removes mutagenic, carcinogenic, and cytotoxic adducts from O6-methylguanine in DNA through a direct reversal mechanism. Decreased expression of MGMT has been reported in a variety of human malignant tumors. The purpose of this study was to clarify the correlation of MGMT expression levels i...
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Isolation, Characterization and Identification of Endophytic Bacteria in Maize (Zea Mays L.) Cultivated on Acrisols of the Southeast of Vietnam
Dang Thi Ngoc Thanh,
Cao Ngoc Diep
Issue:
Volume 2, Issue 4, August 2014
Pages:
224-233
Received:
11 August 2014
Accepted:
19 August 2014
Published:
30 August 2014
Abstract: Endophytic bacterial diversity in Maize plant cultivated on Acrisols of the Eastern of South Vietnam was studied. Maize material was collected from five sites (provinces/cities) of this region. Endophytic bacteria were isolated in two kinds of medium (LGI, NFb) together with 16S rRNA gene fragments amplified from DNA using eubacterial universal primers (p515FPL and p13B). A total of 301 isolates were isolated and all of them have ability of nitrogen fixation and phosphate solubilization together with IAA biosynthesis but there were 30 isolates having the best characteristics and they were identified as maize endophytes and nifH gene owners. The sequences from selected endophytic bacteria (30 isolates) showed high degrees of similarity to those of the GenBank references strains (between 97% and 100%). Among the selected isolates were 6 isolates belong to Bacillus (20.00%) and 24 isolates belong to Proteobacteria (80.00%) including 2 alpha-proteobacteria (7%), 6 beta-proteobacteria (20%), and 16 gamma-proteobacteria (53%). Based on Pi value (nucleotide diversity), Proteobacteria group had the highest Theta values and Theta values (per sequence) from S of SNP for DNA polymorphism were calculated for each group and Proteobacteria group had the highest values in comparison of two groups. From these results showed that 3 isolates including DTN1b (Azotobacter vinelandii), VTN2b (Bacillus subtilis) and VTN7 (Enterobacter cloacae) proposed as potential microbial inoculants or biofertilizers for sustainable corn production in poor Acrisols in Vietnam because of their benefit and biosafety.
Abstract: Endophytic bacterial diversity in Maize plant cultivated on Acrisols of the Eastern of South Vietnam was studied. Maize material was collected from five sites (provinces/cities) of this region. Endophytic bacteria were isolated in two kinds of medium (LGI, NFb) together with 16S rRNA gene fragments amplified from DNA using eubacterial universal pri...
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TNF-α Genetic Polymorphisms and its Expression in Egyptian Rheumatoid Arthritis Patients
Afaf K. Mousa,
Ahmed M. Ghoneim,
Noha B. El-Mashad,
Ahmed E. El-Ghobashy
Issue:
Volume 2, Issue 4, August 2014
Pages:
234-240
Received:
15 August 2014
Accepted:
23 August 2014
Published:
30 August 2014
Abstract: Rheumatoid arthritis (RA) is a complex, multifactorial, inflammatory disease that affects more than 1.5 million adults. The current study aimed to investigate whether there is an association between either -376 G/A or -238 G/A polymorphisms of TNF-α promoter and the risk of RA in Egyptian patients, and investigate whether these polymorphisms are linked to TNF-α expression. DNA from 54 clinically confirmed RA patients and 24 apparently healthy individuals was genotyped by RFLP technique. Some samples were selected for semi-quantitative measurement of TNF-α mRNA expression. For the -376 polymorphism, no polymorphism was recorded in this site neither in RA patients nor in the apparently healthy individuals. This indicated the wide distribution of the wild type GG genotype among Egyptians. For the -238 G/A polymorphism, data indicated that 77.8% of RA patients were of the genotype GG and 22.2% were heterozygous (GA), while 91.7% of the apparently healthy individuals were of the genotype GG and 8.3% were heterozygous (GA). The homozygous genotype AA was not recorded in any RA or healthy subject. There was no statistically significant difference in the genotype distribution between RA patients and the apparently healthy individuals. Also, there was no statistically significant difference in either the G or A allele distribution between the RA group and the group of healthy subjects. Semi-quantitative PCR on some samples revealed a statistically significant increase in the relative expression of TNF-α mRNA in RA patients compared to healthy subjects. Based on these data, we conclude that -238 G/A and -376 G/A polymorphisms can not be considered as risk factors for RA among Egyptians and the increased expression of TNF-α in Egyptian RA patients is not linked to these polymorphisms. Therefore, Egyptian RA patients may have different genetic or environmental factors contributing to the pathogenesis of RA and further studies are necessary to search for other genetic polymorphisms and/or genes that contribute to the increased expression of TNF-α and hence the pathogenesis of RA in Egyptian patients.
Abstract: Rheumatoid arthritis (RA) is a complex, multifactorial, inflammatory disease that affects more than 1.5 million adults. The current study aimed to investigate whether there is an association between either -376 G/A or -238 G/A polymorphisms of TNF-α promoter and the risk of RA in Egyptian patients, and investigate whether these polymorphisms are li...
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Ramipril and Valsartan as Protective Agents against Some Complications of an Experimentally Induced Obesity in Rats
Said Al-Dalaen,
Aiman Al-Qtaitat,
Samir Mahgoub,
Nedal Alnawaiseh,
Jehad Al-Shuneigat,
Samih Sarayreh,
Yousef Sarayreh
Issue:
Volume 2, Issue 4, August 2014
Pages:
241-250
Received:
3 August 2014
Accepted:
27 August 2014
Published:
10 September 2014
Abstract: Objectives: Our work aimed to study the effect of experimentally induced obesity on the rate of advanced glycated end products (AGEs) formation and the activity of angiotensin converting enzyme (ACE) and its relation to oxidative stress and also to evaluate the protective effect of ramipril (an angiotensin converting enzyme inhibitor), valsartan [an angiotensin II blocker; AT1 receptor blocker), and their combination on these obese animals. Materials and Methods: The present study was conducted on ten female albino rats fed on standard chow as a control group and fifty obese animals received for sixteen weeks high fat diet alone or in concomitant combination with either ramipril (2 mg/kg/day or 0.25 mg/kg) or valsartan (0.30 mg/kg/day) or the combination of both drugs (0.25 mg/kg of ramipril and 0.30 mg/kg of valsartan daily for sixteen weeks). Blood, kidney and aortic AGEs, ACE activity and advanced oxidation protein products (AOPPs) were measured. Results: The obtained results showed increase in triacylglycerols (TGs) levels (p<0.043) in the obese animals versus the control group. The total blood cholesterol (TC) and LDL-cholesterol (LDL-C) levels, also, were significantly higher (p<0.0001) in obese animals compared to the corresponding values in controls, with a significant reduction in their levels in all treated groups except in group IV (p=.041) when compared to the control group. On the other hand, HDL-cholesterol (HDL-C) was significantly lower (p<0.0001) in the obese animals compared to its level in the controls. The obese animals showed significant increase in their blood glucose and serum insulin levels when compared to the controls [(p<0.037) and (p<0.045), respectively]. The results, also, revealed that obesity was associated with a statistically significant increase in the blood, kidney and aortic tissue levels of AGEs, ACE and AOPPs compared to their corresponding values in the control group. Treatment with ramipril, valsartan and their combination caused significant reduction in serum and tissue levels of both AGEs and AOPPs when compared with the obese group. On the other hand, ACE activity was markedly reduced following the administration of ramipril alone or when it is combined with valsartan, while the administration of valsartan alone showed no significant effect on the activity of ACE when compared to the obese group. Moreover, combination of ramipril (at a submaximal antihypertensive dose of 0.25 mg/kg/day) with valsartan produce a marked reduction in all parameters examined compared to valsartan alone. Conclusion: combination of ramipril and valsartan showed more therapeutic effect compared to individual therapy with ACE inhibitor or AT1 receptor blocker.
Abstract: Objectives: Our work aimed to study the effect of experimentally induced obesity on the rate of advanced glycated end products (AGEs) formation and the activity of angiotensin converting enzyme (ACE) and its relation to oxidative stress and also to evaluate the protective effect of ramipril (an angiotensin converting enzyme inhibitor), valsartan [a...
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