Annual Research & Review in Biology

Antiseptics and disinfectants are important substances for medical care. However, the needs for new antiseptics have increased in the last few years, due to multi-drug resistant pathogens. In this study we aimed to evaluate, the efficacy of 2-aminoindanbiguanide chloride (2-AIB.HCI) and 2-aminoindanbiguanideacetate (2-AIB.HOAc) salts that were synthesized against multi-drug resistant Gram-negative bacilli (MDR-GNB) (Acinetobacter spp, Escherichia coli and Klebsiella pneumoniae). According to our resultsat70 mg/mL concentrations of 2-AIB.HCI and 2-AIB.HOAc salts have been confirmed to be effective against hospital infection causing MDR-GNB. It was shown that 2-AIB.HCI and 2-AIB.HOAc salts have antibacterial efficacy against MDR-GNB.

Aims: The aim of this work is to assess the influence of temperature (T) and precipitation (P) on Basal Area Increment (BAI) of Pinus cooperi in a mixed conifer forest ecosystem. We hypothesize that the sensitivity of annual tree growth to climate variables will increase with increasing elevation.
Study Design: The study area is located in "Las Rusias" mountain, in northwest Mexico. The sample design consisted of selection of three sites with different elevations: Low (L), Mid (M) and High (H), of representative stands of Pinus cooperi.
Place and Duration of Study: The field work was carried out in October 2011, whilst processing data was done at Lab in December 2011.
Methodology: Increment cores were collected from 3-10 trees at each site along the elevational gradient. Increment cores were extracted for each tree of Pinus cooperi at 1.3 m height using increment borers of 5 mm. Increment cores were processed using standard dendrochronological techniques. In order to determine the climatic response of the BAI chronology, BAI data was associated with climate records (1946-2010) from the nearby weather station.
Results: The bootstrapped procedure conducted between the BAI and climatic variables showed significant correlations for the study area. All correlation coefficient values ranged from -0.5 to 0.42 (p<0.05). The Pearson´s coefficients indicated that BAI has been driven largely by (P) and (T) along the gradient.
The relationships between BAI and P showed similar trends for the three elevations. P. cooperi BAI was positively correlated with precipitation during the previous winter.
Conclusion: The results showed that the hypothesis that sensitivity to rainfall and temperature will vary along the elevation gradient appears reasonable. These findings could be used to suggest strategies for forest management, however some stand dynamics factors should be considered as well.

Introduction: The foramen magnum (FM) is a unique and complex anatomical area located in the central basal region of the occipital bone. It is a three-dimensional opening through which vital structures pass, such as the medulla oblongata and its membranes, the spinal accessory nerve, the vertebral arteries, the anterior and posterior spinal arteries, the tectorial membrane and the alar ligaments. Because of this vital relationship between the FM and its content, it is of great importance to study the morphological types of FM. Findings from such evaluations have essential applications in clinical practice and in surgery.
Objective: To determine the incidence of different morphological types of FM.
Materials and Methods: 110 FM from dry human crania of known sex and age were analyzed. The specimens belong to the anatomical collections of Tiradentes University (UNIT) and the Metropolitan University of Education and Culture (UNIME). Among these, 66 were male, 44 were female and their ages ranged from 11 to 91 years (mean: 58.01 years). FM morphology was studied using photographic images and with the aid of Corel Draw Photo-Paint X5.
Results: Among the 110 FM analyzed, nine morphological types were found: pear-shaped 37.27%, rounded 15.45%, tetragonal 10.91%, biconvex 10.91%, hexagonal 9.09%, oval 5.45%, pentagonal 2.73% and heptagonal 1.82%. In the remaining 6.36% of the FM, it was not possible to define a regular and specific form.
Conclusions: Pear-shaped FM predominated, followed by rounded FM. In relation to sex, the tetragonal and biconvex forms occurred five times more frequently in males than in females, while the hexagonal form occurred twice as frequently in females than in males.

Herbal plants have been found to be effective in traditional medicine for healthcare. However, the full potentials of these natural resources are yet to be fully explored. This research was coined at investigating the effects of Tetracarpidium conophorum seeds on the hormone and sperm profile of male albino rats. Forty eight albino rats of about twelve weeks weighing between 130-180g each were divided into four groups (A, B, C and D) with twelve rats in each group. The test extract was obtained from the seeds and incorporated into the feed of the rats. Group A served as the control (without test substance) while groups B, C and D were fed with of 4, 8 and 12g/kg body weight (BW) of the test substance for the period of 63 days. Respectively the results obtained from hormonal analysis show that there are significant difference (p<0.05) in the serum level of follicle stimulating hormone (FSH) and luteinizing hormone (LH) of the rats between the different treatment groups while the serum levels of testosterone and estradiol were not significant (p>0.05). Results of semen quality show that there are significant differences (p<0.05) in the sperm count, sperm morphology, sperm viability and semen pH among rats between the different groups. Sperm motility and organ weights (testes and epididymides) were not significantly affected (p>0.05). The results indicate that the seeds of T. conophorum can enhance the production of reproductive hormones and may be used in the formulation of useful fertility drugs.

Passion fruit is of high economic importance in Kenya. In the recent past, diseases have led to decline in fruit production. This study aimed at identifying disease management practices that could be adopted to mitigate disease impacts. Passion fruit varieties used include purple passion (Passiflora edulis Sims) and the KPF hybrid. Field maintenance practices assessed include frequency of orchard sanitation through regular weeding and pruning, pesticide application, nutrient and water provision to plants during the dry season. Agronomic factors assessed include grafting, prunning and effect of wounding in increasing plants’ susceptibility to dieback infections. Assessments were done on-farm and on station; 3 research sites were identified in the field and 1 site was set up on-station. For each of the 3 on-farm sites, a farmer’s performance in orchard maintenance was rated for all the factors on a score of 1-5 and compared to disease severity (1-5 dieback scale ) and incidence (0-100%). Experiments were set up to assess role of grafting on 4 months old purple passion seedlings, while effects of prunning and wounding were assessed on 9 months old passion fruit plants of both varieties. Disease establishment was observed and severity was assessed based on the 1-5 dieback scoring chart. Results showed that proper field maintenance reduced disease incidence and severity (12% and 1.6, respectively) in site 1, while poor maintenance yielded higher levels (55% and 4.0, respectively) in site 3. Agronomic practices important in transmission of dieback pathogens on the purple variety were pruning, grafting and wounding. These were found to enhance disease establishment and severity (P ≤ 0.05) on the purple variety while KPF 12 was tolerant to dieback infections. The study established that proper agronomy and field maintenance practices are important and should be integral in control of dieback disease.

This study examined the effects of chilling, freezing, packaging, temperature and storage time on the physical, chemical changes and microbiological growth in barred Spanish mackerel (Scomberomorous commerson). At time 0 the free fatty acid, trimethylamine, and thiobarbituric acid reactive substances (TBARS) valueswere1.2%, 0.30 mg- N/100g and 0.17mgmalonaldehyde/kg, respectively. Total viable cell count and coli forms fecal were 4.6 and 3.3 log₁₀ CFU/g, respectively. Barred Spanish mackerel was chilled at two different temperatures (4 and 7ºC) for6 days. Also, fresh mackerel was kept frozen up to 6 months at –10 and -18ºC. Sampling was carried out on the initial and at 1, 3, and 6 days for chilling and 0, 1, 3 and 6 months for freezing. Physical and chemical changes of the mackerel were evaluated by increasing TBARS values, trimethylamine and free fatty acid. Drip loss and Torry meter were also measured at various time intervals for six days at 4ºC, for three days at 7ºC and for 180 days at –10 and -18ºC. Shelf life was found to be 6 and 3 days for barred Spanish mackerel stored at 4ºC and 7ºC, respectively. Stored barred Spanish mackerel either at -10 and -18ºC showed longer acceptability tolerance (180 days). Generally, increasing temperature from 4 to 7ºC has led to about 50% decrease in the freshness period and shelf life. Freezing storage allowed the retention of freshness qualities of barred Spanish mackerel for extending their shelf life and the improved quality.

Aims: Antibiotic resistance is one of the widely recognized public health challenges in Bangladesh. The present study was aimed to analyze the current status of irrational use of antibiotics in rural Bangladesh and to explore the views from both physicians’ and patients’ perspective.
Study Design: Population based survey.
Place and Duration of Study: The survey was conducted among 6,000 patients and 580 physicians in the rural areas of Dhaka and Rajshahi divisions of Bangladesh from July 2012 to December 2012.
Methodology: The survey followed a face-to-face interview protocol. 24 Upazila Health Complexes and 112 Union Health Centers of Dhaka and Rajshahi divisions were conveniently surveyed by trained volunteer interviewers. Two separate survey questionnaires were developed for physician and patient survey.
Results: From the physician survey it was found that significantly more doctors prescribe antibiotics in suspected infections (P<.0001). Around forty-four (44.1) percent doctors prescribe antibiotics in cold and fever before diagnosis. A significant proportion of physicians never receive any feedback about the antibiotic they prescribe (31.9%, P<.0001) and more than 50% doctors claimed that they receive feedback occasionally, not always (P<.0001). According to the physician’s patient non-compliance is the main cause of antibiotic resistance in the country (68.8%). Though 48.6% patients think that it is important to strictly follow the doctor’s prescription, a significant percentage believe that it is not always necessary (26.7%, P<.0001) and more than 50% patient stop taking the antibiotic as soon as the symptoms disappear, while only 25.2% patient complete their full course. Only 6.3% patients consult their doctor if they miss the dose of an antibiotic and more than 50% take the next dose on time (P<.0001). When a drug does not work the patient usually consider the doctor is incompetent (25.6%) and many (24.5%) believe that the quality of the drug is not up to the mark.
Conclusion: The result of this survey indicates that the antibiotics are used among the rural people in irrational way. To overcome this situation close supervision of the relevant authority is required in order to minimize the growing antibiotic resistance in Bangladesh.

Aims: Behavioral responses of 3-month-old male pups from female Wistar rats exposed to daily 1 hour or 3 hour restraint stress during the first 7 days of pregnancy were studied by tail flick and formalin test.
Methodology: Eighteen mature virgin female albino rats (140g-160g) were randomly allocated in a blinded fashion to 3 groups (n=6 each) and mated. Group 1 rats were the control and did not undergo restraint stress. Groups 2 and 3 rats were restrained for 1 hour and 3 hours respectively during the first 7 days of pregnancy. Six male rats only served purpose of copulation. At 3 month of age, 34 pups consisting of 10, 11 and 13 pups delivered by rats in groups 1, 2 and 3 respectively were randomly selected and studied for nociception.
Results: Body weights were higher in both 1 hour and 3 hour prenatally stressed pups compared to that of the control. The latency period during the tail immersion test in the pups prenatally stressed for 3 hour daily but not those stressed for 1 hour daily was significantly higher (p<0.05) compared to that of control. While there were no significant differences in the formalin score in pups prenatally stressed for 1 hour and 3 hour compared to the score of the control during the early phase, the formalin score of the pups prenatally stressed for 3 hour daily but not those stressed for 1 hour daily was significantly lower compared to that of the control during the late phase.
Conclusion: Our findings suggest that early prolonged prenatal stress modulates nociceptive sensitivity in 3-month-old rat and that different mechanisms are responsible for the effects of prenatal stress on acute and persistent pain in the formalin test.

Aims: To evaluate the effect of garlic oil and turmeric extract on rumen fermentation and methane emission using an in vitro gas production technique.
Study Design: Completely randomized design.
Place and Duration of Study: Department of animal science, Faculty of agriculture, Ferdowsi University of Mashhad, between November 2012 and June 2013.
Methodology: Treatments were: control (no additives); 10 mg/L of cultural fluid of monensin (MO) as a positive control; 20, 40 and 80 mg/L of cultural fluid of garlic oil (GA20, GA40 and GA80 respectively); 20, 40 and 80 mg/L of cultural fluid of turmeric extract (TU20, TU40 and TU80 respectively). All treatments were incubated for 96 h and gas production was recorded by a pressure transducer at 2, 4, 6, 8, 12, 24, 48, 72 and 96 h of incubation. At 24 h post incubation, pH of cultural medium and methane production were determined.
Results: Supplementation of MO and GA80 resulted in a significant reduction in total gas production compared with the control. Furthermore, addition of garlic oil at 80 mg/L strongly decreased methane production during 24 h post incubation (43% lower than the control). Mid and high concentrations of turmeric extract (TU40 and TU80) caused to increase in vitro dry matter disappearance (IVDMD) in comparison with the control. Partitioning factor (PF = mg DM disappeared/ ml gas produced after 24 h incubation) and pH of the cultural medium were higher using MO, TU20 and GA80 compared with that of the control. Lower extent of gas production (b) and gas production rate (c) were observed in MO and GA80 compared with those of the control.
Conclusion: Present data indicated that using natural additives might alter rumen fermentation pattern as observed in methane emission from a mixed diet.

Aim: While antioxidant effect of ginger rhizome in reducing the oxidative stress in many diseases is well reported, there are limited studies on its effect in improving testicular function of cryptorchid rats. The present study was therefore carried out on cryptorchid rats given ethanol extract of ginger rhizome (EEG) to evaluate the effect on epididymal sperm and plasma oxidative stress parameters.
Methodology: Twenty-four male Sprague-Dawley rats (170g-210g) were randomly divided in a blinded fashion into 3 groups (n=8). Group A was sham-operated and treated with vehicle (corn-oil). Groups B and C were rendered cryptorchid and treated with vehicle and EEG respectively.
Results: Cryptorchid rats demonstrated significant decline in testicular weight leading to reduced epididymal sperm count, motility and percentage of morphologically normal sperm. Plasma oxidative stress was evident as there was rise in malondialdehyde (MDA) levels but decrease in the activity of the antioxidant enzyme superoxide dismutase (SOD). EEG treatment however, significantly (p<0.05) improved all the above parameters.
Conclusion: The above findings indicate that EEG treatment helped to improve the testicular function in cryptorchid rats probably by counteracting the rise in oxidative stress in the plasma.

Many species of protozoa present in the alimentary canal of human beings causing diseases. Due to water-borne diseases about 3.5 million people including 3 million children die throughout the world as well as about 98% deaths occur through extensive water-borne outbreaks in the emerging republics Only the diarrheal diseases cause greater than 1.5 million deaths per year. To investigate the presence of E. histolytica in different water sources of district Peshawar. The study was designed for molecular detection of E. histolytica in water sources. A total of 300 water samples were collected from different water sources of district Peshawar from May, 2011 to April, 2012. And for further process the samples were brought to the Department of Zoology Kohat University of Science and Technology, Kohat on time. Water samples (n=300) were collected from different water sources (Tube Well, Bore Well, Tap and Drain) in six different areas of District Peshawar (Pakistan). The water was filtered thorough whattman filter paper and the residue was subjected to DNA extraction and PCR was conducted for detection of E. histolytica. To increase the sensitivity of the test a small region (125-bp) of the SSU rRNA was targeted for the PCR amplification. pH of the water were also tested, mean value for over all pH was 8.21 (±0.06), including 8.16 (±0.20) of tube well water, 8.30 (±0.32) of bore well water, 8.26 (±0.24) of tap water and the pH of drain water was 8.11 (±0.48). Overall prevalence of E. histolytica in drinking water of district Peshawar was 11.33% (34/300) followed by 3.57% (2/56) in tube well, 2.74% (2/73) in bore well, 14.41% (16/111) in tap water and 23.33% (14/60) in drain water. The highest prevalence of E. histolytica was 65% recorded in tap water of Faisal Colony and P<.05 was considered significant. It was revealed from the current study that E. histolytica is present in water sources of some areas in Peshawar which may be due to flood and improper management of water scheme. The study also revealed that a proper treatment of water for human consumption is required especially in Faisal Colony in district Peshawar.

Aim: There are differences in the frequency of CYP2C19 mutant alleles among different ethnic group. The aim of present study was to estimate the distribution of CYP2C19 allele and genotypic variants in Iranian Fars ethnic group and compare it with other populations.
Study Design: Polymerase Chain Reaction (PCR)-Restriction Fragment Length Polymorphism (RFLP) technique was used to determine Genotyping of CYP2C19 alleles.

Place and Duration of Study: Biochemistry and Biophysics, Metabolic Disorders Research Center, Gorgan Faculty of Medicine, Golestan University of Medical Sciences, between September 2012 and August 2013.
Methodology: To determine the genotype and allelic frequency of CYP2C19, 140 unrelated healthy Fars origin people who were referred to Health Center, were included this study.
Results: The allele frequency of CYP2C19*1, CYP2C19*2 and CYP2C19*3 were 77.8%, 19.2% and 2.8%, respectively. 75% of subjects were with CYP2C19*1/*1 genotype. 22.1%, 1.4%, 1.4%, 0% and 0% subjects were with CYP2C19*1/*2, CYP2C19*1/*3, CYP2C19*2/*2, CYP2C19*2/*3 and CYP2C19*3/*3 genotypes, respectively. Poor metabolizer, intermediate metabolizer and extensive metabolizers genotype frequencies were seen in 75%, 23.5% and 1.4% of subjects, respectively.
Conclusion: Ethnic differences in CYP2C19 genetic polymorphism of cytochrome p450 enzyme may cause variation in drug response, activity or detoxification. This study suggests the further study of this polymorphic enzyme in Fars population to determine the clinical significance and optimal dosage of some drugs in this ethnic group.

Aims: The aim of this research was to determine the chemical and fatty acids composition of different genotypes of tobacco seeds.
Place and Duration of Study: Ten tobacco genotypes grown in Sulaimani-Iraq were investigated in this study. The tobacco seeds were provided by Agricultural station of Bazian, Ministry of Agriculture. The investigations were carried out on air-dried seeds during 2013 in University of Sulaimani, Faculty of Agricultural Sciences.
Methodology: Plants were sampled at different locations of Sulaimani to determine the chemical and fatty acid composition. The seed chemical and fatty acids compositions were determined by Soxhlet, Kjeldahl, Column chromatography, TLC and Gas chromatography methods.
Results: There were significant differences (p˂ 0.05) among genotypes for all studied chemical composition. Protein, ash, fiber and oil contents ranged from 20.861 to 23.872%, from 2.067 to 3.467%, from 13.66 to 19.33% and from 24.56 to 41.933% respectively. The results obtained from phospholipid, sterols and tocopherol showed significant difference among all genotypes. The content of phospholipids, sterols and tocopherols in the oils was 0.453-1.167%, 0.2-0.373% and 0.005-0.007% respectively. The results showed significant difference (p˂ 0.05) among genotypes for all saturated and unsaturated fatty acid. Palmitic saturated fatty acid (21.33 to 25.667%) and oleic unsaturated fatty acids (17.00 to 26.667%) were predominant in the oils.
Conclusion: The results indicated that all genotypes contained large amount of lipid, protein and fiber. Our data showed that the amount of unsaturated fatty acids is higher than the amount of saturated fatty acids in tobacco seeds. The knowledge of the present studies on different genotypes of tobacco seeds could be important to its appropriate industrial use and for improvement in the nutritional value.

Aims: To investigate the antimicrobial activity of Phomopsis sp. ED2, endophytic fungus isolated from Orthosiphon stamineus Benth against various pathogenic microorganisms.
Study Design: Place and Duration of Study: Industrial Biotechnology Research Laboratory, Universiti Sains Malaysia, Penang, Malaysia, between Dec 2011 to August 2013.
Methodology: The endophytic fungus Phomopsis sp. ED2 was cultivated in the culture medium supplemented with the host plant extract. Then the antimicrobial activities of the crude extracts were screened via disc diffusion assay. The minimal inhibitory concentration and minimal lethality concentration of the extract against selected test microorganisms by using broth microdilution assay. The bioactive constituents of the extract were analyzed by using gas chromatography-mass spectroscopy analysis.
Results: The ethyl acetate extract of this fungus showed broad spectrum inhibitory activity on both Gram positive and Gram negative bacteria with the minimal inhibitory concentrations ranged from 31.25 to 250 µg/mL. The gas chromatography-mass spectroscopy analysis of the extract showed that the bioactive constituents present in the extract were benzoic acid, lomustine and penicillic acid.
Conclusion: The endophytic fungus residing in Orthosiphon stamineus Benth, Phomopsis sp. ED2 exhibited significant antibacterial activity of the test microorganisms. It is a potential source of antibacterial compounds.

There are many barriers that have hindered the progress of clinical application of gene therapy as a potential method for treating cancer. These limitations are caused by transduction/delivery failure rates with viral vector systems and were due to the resultant non-specific targeting as well as the triggering of immune system stimulation. Recently, mesenchymal stem cells (MSCs) were found to be advantageous in improving tumor tropism as well as in providing an immune-privilege. Therefore, the combination of viral vector systems and stem cells was suggested as an attractive solution to promote targeted-delivery of anticancer therapeutics to tumor sites. In this regard, many published studies have shown that MSCs could be useful as a potential vehicle for delivering anticancer agents to tumors. In this review, the disadvantages of viral vector systems for gene therapy were analyzed, and an updated account on the role of MSCs in cancer gene therapy was presented. In addition, major safety and therapeutic unresolved problems facing this approach in clinical application were also addressed.