Annual Research & Review in Biology

Aims: The aim of the present study was to investigate some biochemical changes in field grown lettuce (Lactuca sativa L. cv Romaine) plants in terms of importance of the accumulation of anthocyanins, flavonoids and photosynthetic pigments as well as photosynthetic limitations which changed during exposure of plants to drought stress and UV-B radiation in order to circumvent the deleterious effects of these Stresses.
Place and Duration of Study: The experiment was conducted under filed conditions from November 2012 to January 2013, at the Agricultural Research Center, KAU.
Methodology: The experimental design was a factorial arrangement in randomized complete blocks with four replicates. The first factor was UV-B (300 nm). The second factor was irrigation regime (complete irrigation to field capacity and limited irrigation. Gas exchange measurements were carried out using a LI-6200 portable IRGA. Chlorophyll fluorescence of Fv/Fm was measured by PAM 2000 fluorometer. Biochemical analyses and antioxidant enzymes assays were performed according to the appropriate methods.
Results: Exposure of lettuce plants to enhanced UV-B radiation and drought stresses (DS) negatively and significantly affected the process of photosynthesis including CO₂ assimilation (P_N), stomatal conductance to water vapour (g_s) and transpiration rate (E). However, the amplitude of the effects of both stressors was dependent on the interactions. This resulted in alleviation of the negative effect of drought on photosynthesis and transpiration by UV-B radiation as the water stress intensified. Intercellular CO₂ (C_i) concentration was only reduced due to water stress compared to control plants. The maximum efficiency of photosystem II (F_v/F_m) was not affected by UV-B radiation stress but reduced by drought. There was an increase in the activities of some antioxidant due to both stresses when applied singly and in combination UV-B irradiation increased the contents of the UV-B absorbing compounds (carotenoids, soluble phenols, anthocyanins), while drought stress caused a notable increase in free proline content. Increase in content of Proline may be the drought-induced factor which plays a protective role in response to UV-B.
Conclusion: UV-B radiation provoked in general more severe damage, evaluated as changes in the amounts of stress markers, than DS, when applied separately. Under multiple stress conditions, each of the stress factors seems to bring out some adaptive effects to reduce the damage experienced by plants caused by the other one. DS can induce accumulation of UV-B absorbing compounds (flavonoids, carotenoids and soluble phenols), which is likely to offer some increased protection from UV-B.

Recent studies have shown that wild birds may act as potential barometers of environmental pollution. We studied the effects of two environmental toxicants, cadmium (Cd) and paraquat (PQ), on the skin fibroblasts of three wild birds including, green-winged teal (Anas carolinensis), bobwhite quail (Colinus virginianus), and house sparrow (Passer domesticus) of the orders Anseriformes, Galliformes and Passeriformes, respectively. The toxicities were represented as 50% lethal concentrations (LC₅₀), a dose causing death of 50% of the cells after 24 h exposure of toxicant. The LC₅₀ of Cd for the skin fibroblasts from green-winged teal, bobwhite quail, and house sparrow were found to be 0.015 mM, 0.078 mM, and 0.176 mM. The LC₅₀s of PQ for the same cells were 4.58 mM, 25.32 mM, and 36.53 mM, respectively. These results indicated that cells from water fowl may be highly susceptible to Cd and PQ poisoning whereas house sparrow was the least susceptible. Further studies are warranted to investigate the mechanism of differential susceptibility of birds to environmental toxins.

Aim: The present study was conducted to explore the allelic and genotypic frequencies of CYP1A2 gene polymorphisms, CYP1A2*1C (−3860 G>A) and CYP1A2*1F (−163 C>A), among 200 healthy Iranian volunteers.
Study Design: Completely randomized design.
Place and Duration of Study: Place – Biotechnology Department, School of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran. Duration – June, 2011 to May, 2013.
Methodology: DNA was extracted from the blood samples by salting-out method. Primers and probes were designed by the Primer Express® Software v3.0.1. Allelic discrimination of the CYP1A2 alleles was performed by Real-Time PCR, and the data were analyzed by the SPSS 16.00 software.
Results: The frequency of each polymorphism in Iranian population was found to be 0.05 and 0.3 for CYP1A2*1C and CYP1A2*1F, respectively. Our data did not show any difference between the Iranian, Turkish, and Egyptian populations in CYP1A2*1C polymorphism; also, CYP1A2*1F polymorphism analysis did not show any differences between Iranian, Turkish, Egyptian, Chinese, Serbian, and Japanese populations. Allele frequency in our population was in equilibrium according to Hardy–Weinberg principle (P=0.05).
Conclusion: This is the first study analyzing the CYP1A2 clinically important allelic variants in Iranian population. Our results indicate that functional significance of these alleles should be evaluated in various populations. It is necessary to determine the relationship between these variants and CYP1A2-related drug metabolism in associated groups.

Aims: The aim of our study was to isolate and characterized the potential fungal species, which can be used for future rehabilitation of these environment.
Study Design: Cross-sectional study.
Place and Duration of Study: Calancan Bay, Marinduque, between November 2011 and December 2012.
Methodology: Copper-laden sediments were obtained in the Calancan Bay; cultured and fungal species were examined morphologically by both macroscopic and microscopic means. Edaphic factors (soil temperature, moisture, pH, organic matter, and texture) were also determined in all randomly selected stations.
Results: Five fungal genera were isolated from the Calancan Bay, namely, Aspergillus sp., Penicillium sp., Fonsecaea sp., Trichoderma sp. and Fusarium sp.
Conclusion: The diversity and occurrence of the isolated fungal species were dependent on the edaphic factors emanating in the area.

Aims: Wheat performance under both Irrigated and dry land areas is affected by salinity and decreased. Like other plants, wheat cultivars try to indicate salt resistance through biochemical, physiological and morphological changes. In this study, responsive proteins in wheat leaves under salt stress were investigated by proteomics technique.
Study Design: Factorial experiment based on a randomized complete block design
Methodology: In this research, tolerant (Roshan) and sensitive (Ghods) cultivars were treated by 200mM salinity during 17 days. A proteomic technique was used to study proteins of leaf. Proteins of the leaves were extracted by TCA–acetone, and separated by two-dimensional gel electrophoresis at pH 4–7.
Results: 200 repeatable spots were identified by using Melanie software and analyzed statically. 24 and 12 responsive spots were identified by using MALDI TOF-TOF mass spectrometry in tolerant and sensitive cultivars respectively. 5 responsive proteins were overlap between both cultivars. In tolerant cultivar, Expression of 19 spots was increased and expression of 5 spots was decreased. All the identified proteins are involved in the regulation of carbohydrate, energy and nitrogen metabolism, photosynthesis related proteins, ROS scavenging proteins, signaling related protein, transporters and chaperon. But, all identified proteins except one of them in sensitive cultivar were down regulated under salt stress.
Conclusion: Based on this study, salt tolerant cultivar could alleviate salt stress. But salt sensitive cultivar couldn’t tolerate salinity and it seems that salt stress have destructive effects on its proteins. In other words, protein compositions of salt sensitive cultivar are more deeply damaged by salt stress than salt tolerant cultivar and protein decay could be a marker of the sensitivity of the concerned cultivar.

Aims: This work is aimed at toxicological effects of three Bacillus species employed as bio control agent against mosquito larvae in terrestrial environment.
Study Design: Using animal model where weight, hematology and histopathology were considered.
Place and Duration of Study: At Microbiology Department, Federal University of Technology, Akure, Nigeria, between December 2009 and February, 2010.
Methodology: Twenty wistar rats weighing 180±0.61g were used and fed with mice feed and water ad libitum for one week to be acclimatize before dividing into four groups of five. Group 1 was the control and was allowed to normal rat feed and water only, group II – IV were subjected to daily single subcutaneous injection with 200ï­l of 10⁷cfu/ml cell dose of Bacillus thuringiensis, Bacillus subtilis and Bacillus cereus respectively for seven days. The rats were observed shortly after each dosing and thereafter twice daily for one week for general behavioral signs of toxicity and possibly mortality.
Results: The rats infected with B. cereus lost weight by 3.8% during the period of study while those in the control, the treated with B. thuringiensis var. isrealensis and B. subtilis gained weight by an average of 3% of the initial weight. In the hematological analysis of the rats, there was significant difference (P<0.05) in values when compared with the control group. Though decrease in differential counts was observed in the rats injected with B. subtilis and B. thuringiensis, significant difference (P<0.05) in values was not observed with the control group. Histopatological examinations of the liver and small intestine of the rats treated with B. cereus had observable pathological changes when compared with control, B. thuringiensis and B. subtilis treat groups.
Conclusion: There were no significant toxicological changes in the groups treated with B. subtilis and B. thuringiensis when compared with the control group. Based on these observations, it could be concluded that these bacterial species could be used as bio control agent in human environment without causing havoc to humans and animals.

Context: Telfairia occidentalis Hook (Cucurbitaceae), a popular vegetable in West Africa, has been reported to have antidiabetic property. But the active components are not yet known.
Aims: This study was carried out to determine the hypoglycemic potentials and the antidiabetic property of polysaccharides of the leaf of Telfairia occidentalis.
Place and Duration of Study: Hypertension and Cardiovascular Physiology Research Laboratory; School of Pharmaceutical Sciences, University Sains Malaysia, Penang, Malaysia, between June 2013 and October 2013.
Methodology: The dried plant material was macerated with water and extracted with ethyl acetate to obtain ethyl acetate and aqueous residue fractions. To obtain the crude polysaccharides, the plant material was boiled with water. Ethanol (96%) was added to the extract. The precipitate obtained was successively washed with ethanol (99%), ether and acetone. Crude proteins were obtained by ammonium sulphate precipitation. The various fractions (250 and 500mg/kg) were orally administered to normoglycemic and STZ-induced diabetic rats. Blood glucose was evaluated with a glucometer at 0, 1, 2 and 4 hours, after administration. Glibenclamide (4mg/kg) was used as the positive control. Column chromatography and HPLC analyses of the aqueous residue and crude polysaccharides were carried out.
Results: None of the fractions at the two dose levels affected blood glucose concentration of the normoglycemic rats. However, the aqueous residue and crude polysaccharides significantly (p=0.05) reduced glucose level at 4 hours in the diabetic rats from 100% at 0 hour to 74.28 and 60.87% at 4 hours, respectively. Based on the column chromatography and HPLC profiles, the hypoglycemic activity of the aqueous residue and crude polysaccharides fractions could be attributed to their carbohydrates content.
Conclusion: Polysaccharides from the leaf of Telfairia occidentalis possess hypoglycemic activity and have the potential of being useful in the management of diabetes. Further work needs to be done to characterize the active polysaccharides.

Aims: The aim of this study was to assess the prevalence, pattern and predictors of herbal medicine use among patients with type 2 diabetes.
Study Design: Observational cross-sectional study design.
Place and Duration of Study: This study was conducted in 2012 at primary health care centers (PHCCs) in Khartoum.
Methodology: Data for a cross-sectional study were collected via face-to-face interviews using structured questionnaires at primary health care centers (PHCCs) in Khartoum, Sudan.
Results: From a total of 600 patients who responded, 58% reported the use of herbal medicine. Significant factors associated with herbal medicine use were, gender (P=0.006), education (P=0.001), duration of diabetes (P=0.005) and family income (P=0.019).Gender (P=0.001), education, duration of diabetes and place of birth (P=0.05). The most commonly used herbs were fenugreek, black seed, cinnamon and olive. The proposed motivations for using herbal medicines were ease of access, low cost and reports of a positive experience by others. Family and friends were the main source of information for most patients. About 63.8% failed to inform their doctors about using herbs, and the main reasons given for this were fear of a negative response, doctor did not ask, and unnecessary to inform the doctor.
Conclusion: The prevalence of herbal medicine among diabetics is high. The herbs used are not investigated for safety and efficacy and this point to the urgent need for research in this area. In order to overcome a lack of knowledge among doctors about herbs, we propose the integration of herbal medicine into the current medical curriculum so that in future physicians will be better able to communicate with their patients.

Aims: Dill (Anethum graveolens L.) is a perennial herbaceous plant belonging to the (Apiaceae or Umbelliferae) family, with high essential oil content, and dill leaves and and seeds are used as seasoning. This study was conducted in 2011 and 2012 at Iran County, aiming to determine the effects of biological and chemical nitrogen fertilizers on grain yield, essential oil yield, and essential oil composition of dill.
Methodology: The treatments included Nitroxin bio-fertilizer (0% and 100%), and nitrogen (0, and 100 kg ha−1 (NH₄NO₃)), 50% nitroxin×50 kg ha-1nitrogen and control. A completely randomized block design with four replications was adopted.
Results: Results showed highest essential oil content detected in biological fertilizer and chemical fertilizer. Identification of essential oil composition showed that content of carvone increased with application of Nitroxin biofertilizers.
Conclusion: Results indicated, application of Nitroxin biofertilizers enhanced yield and essential oil content in this plant.

Aims: In two successive seasons, the effect of different types of fertilizers on growth, physiological characteristics and oil constituents of thyme plants was studied.
Methodology: The performance of compost and nitroxin were compared with standard, commercial rates of NPK fertilizers with respect to the growth, chemical composition and essential oil production of Thymus vulgaris L.
Results: In two successive seasons, plants treated by a mixture of nitroxin, compost showed a significant increase in vegetative growth, total N, P and carbohydrate content and essential oil production.
Conclusion: The findings clearly indicate that Nitroxin, and compost could replace conventional NPK fertilizers in the cultivation of thymus, and consequently minimize environmental pollution by these compounds.

Aims: It is now clear that cytokines such as interleukins play a fundamental role in inflammatory processes. Cytokines now offer alternative targets for therapeutic intervention and may be useful as predictive biomarkers of disease. In this study, we aimed to evaluate the relationship between cytokines and spinal mu opioid receptor expression during adjuvant-induced arthritis (AA) in rats.
Methodology: AA was induced by a single subcutaneous injection of CFA into the rats’ hindpaw. Anti-IL-6 and IL- 10 were administered daily during the 21 days of study. Hyperalgesia and edema were assessed on days 0, 3, 7, 14 and 21 by radiant heat and plethysmometer respectively. Spinal mu opioid receptor (MOR) expression was detected by Western blotting.
Results: Daily administration of neutralizing doses of anti-IL-6 caused a significant decrease in serum IL-10 level in AA rats, while treatment with anti-IL10 antibody in the AA rats did not significantly change serum IL-6 level. Anti-IL-10 antibody administration caused a significant increase in paw edema and hyperalgesia, while anti-IL-6 treatment decreased heperalgesia on days 3 and 7 but caused a significant increased in hyperalgesia on days 14 and 21 in AA rats. Administration of neutralizing dose of anti-IL10 antibody in AA rats did not cause significant variation of spinal MOR expression in comparison to IL-6 antibody treatment which induced significant reduction in spinal MOR expression.
Conclusion: Together these results suggest that some of the anti-hyperalgesic effects of serum IL-6 during the chronic phase of inflammation may be mediated by increased IL-10 levels.

Aims: The aim of this study is to investigate the genotoxicity of Methyl Tert-Butyl Ether (MTBE) to Vicia faba L. plants.
Study Design: Experiments were carried out to determine the depressive effect in the mitotic process of Vicia faba L. when treated with different concentrations of Methyl Tert-Butyl Ether.
Place and Duration of Study: Department of Biotechnology and Biology, Faculty of Science, Taif University, between January 2011 and February 2011.
Methodology: Vicia faba L. seeds were germinated in different concentrations of Methyl Tert-Butyl Ether (0.5, 1.0, 5.0, 10 and 15 %). For each concentration, five root tips were transferred to five microscope slides, stained with feulgen technique, covered with cover slip, squashed and observed microscopically. The cytotoxicity and genotoxicity induced by each concentration was compared with the value for the concomitant negative control using t-test.
Results: The trend of the results showed that the higher the concentration of MTBE, the inhibitorier the effect on mitosis with more pronounced chromosomal aberrations. Types of abnormalities revealed the induction of spindle disturbance, stickiness, laggards, fragments, bridges and micronuclei which lead to the loss of genetic material.
Conclusion: The induction of sticky chromosomes and micronuclei indicated that the Methyl Tert-Butyl Ether caused abnormal DNA condensation and inactivated the spindles. Because abnormalities of the cell division process results from the genotoxic effects of environmental chemicals, the Methyl Tert-Butyl Ether has the potential to cause aneuploidy in exposed organisms and adverse human health and environmental effects.

Aims: Reproductive behavior of couples, are important for health authorities for planning and policy making. Periodic national surveys, as well as an annual report are being made by the health administrators in every province.
Study Design: A cross- sectional study.
Place and Duration of Study: Thalassemic research center, Mazandaran University of medical science, between January and March 2012.
Methodology: It was a cross- sectional study with the multi stage random sampling. First, three districts were randomly selected. Half of the households selected from each urban and rural residents. Subjects from families were randomly selected from the list of health centers. Validity of the questionnaire for a structured interview was obtained by expert consultation. Experienced female health workers were educated for the filling the questionnaires. Epidemiologic characteristic of couples was assessed by 10 questions. Couples asked if they have planned a pregnancy in near future. Descriptive statistics were used for reporting data. The P<0.05 was considered statistically significant.
Results: Six hundred and sixty families were studied. Age of the man and wife was 39.7 +/-10.5 and 35.5 +/- 10.7 years respectively. Married year's was 14.6 +/- 10.6 per couple, with the sum of 9618 years. Thirty three (5% CI 95%: 2.7 - 7.3) women were pregnant, 38(6.2% CI 95%: 3.6-8.8%) couples were keen to have a near pregnancy. Almost 22% of women were permanently infertile and 82.7% were using safe contraceptive methods. Twenty one (3.7%), 5(0.9%) of families had illegal and medical abortions respectively. About 70% (469) were using contraceptive methods. Most popular safe method was oral pills (20 %). Among permanent methods the most popular one was tubal ligation (TL).
Conclusion: Still a significant proportion of couples are using unsafe contraception methods. However, use of prenatal diagnosis service became recognized.

The objective of this study is to predict the presence of heart disease with reduced number of attributes using data mining techniques. The term heart disease is related to all diverse diseases affecting the heart. The exposure of heart disease from various factors is an issue which is not free from false presumptions often accompanied by unpredictable effects. Researchers have been using several data mining techniques to help health care professionals in the diagnosis of heart disease. In our work, Classification and Regression Tree (CRT) is proposed to determine the attributes which contribute more towards the diagnosis of heart ailments, which indirectly may reduce the number of tests which are needed to be taken by a patient. The dataset used consists of 270 cases. Originally, thirteen attributes were involved (age, sex, chest pain type, resting blood pressure, serum cholesterol, fasting blood sugar, resting electrocardiographic results, maximum heart rate, exercise induced angina, depression induced by exercise relative to rest, slope, number of vessels colored by fluoroscopy, and exercise thallium scintigraphic defects). To evaluate the performance of the model the sensitivity, specificity, and accuracy are calculated. Comparison with other data mining techniques is presented. The simulation result obtained from the model enables to establish significant patterns and relationships between the medical factors and heart disease.

Acyl-CoA:diacylglycerol acyltransferase (DGAT) is a critical enzyme involved in the last committed step of triglyceride synthesis. DGAT has two known isoforms that are expressed in various tissues being crucial to energy homeostasis in mammals. DGAT inhibits the accumulation of diacylglycerol, a lipid thought to promote insulin resistance, by incorporating them into triglyceride. However, both deletion and overexpression studies targeting DGAT have resulted in decreased diacylglycerol content and caused increased insulin sensitivity. Studies analyzing obese models though have noticed a positive correlation between DGAT and obesity. Thus, it remains elucidated whether DGAT plays a causal role in the development of diet-related diseases or provides protection by preventing accumulation of diacylglycerol. The purpose of this review is to present an overview of DGAT, state factors that may influence its expression, explain the potential consequences of DGAT overexpression or deficiency, and discuss its relative importance to glucose metabolism and energy storage.