Detection of Microsatellite Instability by High-Resolution Melting Analysis in Colorectal Cancer

Background: Colorectal cancer is the third most common cancer worldwide. MSI is a molecular marker of a deficient MMR system and happens in almost 15% of CRCs. Because of a wide frequency of MSI⁺ CRC in Iran compared to other parts of the world, the importance of screening for this type of cancer is highlighted. Methods: The most common MSI detection technique is a fluorescent PCR-based method in which fragments are analyzed by CE. This technique is very time-consuming, difficult, and expensive. We sought to develop and evaluate a proper method with high accuracy, specificity, and sensitivity to screen the MSI⁺ CRC. A HRM analysis procedure is relying on the analysis of the melting curve attributes. Low cost, feasibility, high specificity, and sensitivity are outstanding attributes of HRM analysis. Results: Five mononucleotide microsatellite markers, including BAT-25, BAT-26, NR-21, NR-24, and NR-27, in 25 archival CRC tumor tissue samples were compared with normal tissue adjacent using HRM method. The specificity and sensitivity of BAT-25 with HRM method were 100% compared to CE, while other markers had lower sensitivity. However, when all the markers were considered together, the sensitivity and specificity became 100%. The number of MSI⁺ samples was 56%, which shows a higher ratio than previous Iranian studies. The highest MSI was related to BAT-26 (52%). Conclusion: The HRM method is much simpler and more cost-effective than current MSI techniques, and its sensitivity and accuracy are comparable. Therefore, it can serve as an alternative method in cases where CE is unavailable. Key Words: Biomarkers, Microsatellite instability, Colorectal cancer     

Wide hybridization and introgression breeding in safflower: Effectiveness of different selection methods

Possibility of using interspecific hybridization to increase genetic diversity of drought tolerance in safflower (Carthamus tinctorius L.) and effectiveness of different selection methods is poorly understood. The main objectives of this study were to evaluate the effectiveness of (a) interspecific hybridization to expand genetic diversity in safflower for agronomic traits and drought tolerance in advance generations and (b) different selection methods (direct vs. indirect) for improving seed yield. Interspecific hybridizations were performed between species of C. tinctorius × C. palaestinus (TP), C. palaestinus × C. oxyacantha (PO) and C. tinctorius × C. oxyacantha (TO). Based on the field screening of genotypes in F3 generation, four groups of selected genotypes (derived from combination of direct and indirect selection under both drought and normal environments along with selection based on drought tolerance index (STI)) were evaluated in F4 and F5 generations. Indirect selection for seed yield was significantly more efficient than direct selection. Selection in stress environment resulted to more efficiency than selection in normal environment. No significant difference was observed between direct selection and selection based on STI. The result showed that TP was the best population to combine high seed yield and drought tolerance in safflower.     

Simultaneous selection for seed and forage production in cocksfoot (Dactylis glomerata): application of drought tolerance and susceptibility indices

An experiment was conducted to estimate drought tolerance of some cocksfoot genotypes based on seed and forage yield and related indices (including tolerance index, mean productivity, geometric mean productivity, reduction percentage index, stress tolerance index and stress susceptibility index) and to identify relationships between measured traits. Twenty‐five genotypes were evaluated in the field under control and drought‐stress environments over 2 years. The results revealed a high variation for seed yield and forage yield and the measured indices under both moisture conditions. No correlation was observed between seed yield and forage yield under the control treatment, but this correlation was positively significant under the water‐stress treatment. Drought‐tolerant and susceptible genotypes were identified based on seed and forage yield performance. Genotypes 2, 4 and 7 had high seed yield, while genotypes 7 and 10 had high forage yield under both stressed and non‐stressed conditions and were identified as the most tolerant genotypes in each category. The results also revealed that genotypes 1, 2, 4, 6, 7, 8 and 24 had relatively high seed and forage yield in both control and drought‐stressed conditions. These genotypes can be used for further studies to improve both seed and forage yields concurrently.     

Effects of date palm leaf fiber on the thermal and tensile properties of recycled ternary polyolefin blend composites

This work investigated the effects of date palm leaf fiber (DPLF) content on the thermal and tensile properties; and morphology of compatibilized polyolefin ternary blend. Recycled polyolefin ternary blend consisting of low density polyethylene (RLDPE), high density polyethylene (RHDPE) and polypropylene (RPP) were fabricated at different parts per hundred resin (phr) of DPLF. Maleic anhydride grafted polyethylene (MAPE) was used as compatibilizer to enhance the adhesion between filler and polymer matrix. The composites were prepared using melt extrusion and tests samples were produced via injection molding process. Thermal conductivity results showed that as much as 11 % reduction in thermal conductivity was achieved with the incorporation of 30 phr DPLF. Highest tensile strength was observed with the incorporation of 10 phr DPLF. The elongation at break was reduced with the addition of DPLF due to impediment of chain mobility by the fillers. Initial degradation temperature increased with the addition of DPLF. Hence, it is concluded that DPLF can be used to develop green and thermally insulating composites. It is hoped that the present results will stimulate further studies on the thermally insulative materials based on natural fibers reinforced polymer composites for applications in the building industries.     

Detection of specific hydatid antigens and antibodies in serum and urine of experimentally infected sheep

Immunodiagnostic confirmation of cystic human hydatidosis is frequently required before surgical intervention or of chemotherapy. However, it remains inadequate to detect specific antibodies or antigens in some confirmed cases of echinococcosis. This study was carried out to investigate the accuracy of three different immunodiagnostic tests for detection of specific circulating antigens or antibodies in the serum and urine of 13 experimentally infected sheep. For this purpose, Echinococcus granulosus were collected from small intestine of experimentally infected dogs, and 2000 taenid eggs were orally administered to each of the 13 sheep. There were six other sheep, which were kept as the control group. Biweekly serum and urine samples were collected from all the sheep for 4 months after infection. The sera were subjected to indirect hemagglutination test and the concentrated urine samples were subjected to coagglutination and counter immunoelectrophoresis tests. The results revealed that the sensitivity of these tests in detecting the hydatid antigens in the urine or antihydatid antibodies in the serum of the infected sheep reached their maximum in 12th and 13th week after infection; then it decreased in the following weeks. Examination of the non-infected sheep samples throughout the experiment showed that the aforesaid findings were specific only to the infected sheep. It seems that the appearance of specific hydatid antigen in urine and its antibodies in the serum were simultaneous. Although these tests are highly specific, false negative outcomes were encountered in their detection of cystic echinococcosis. In general, it seems rational to establish some series of diagnostic procedures in order to reveal antibodies and antigen of metacestode in serum and urine of the patients.     

Bone Marrow Stromal Cell Transdifferentiation into Oligodendrocyte-Like Cells Using Triiodothyronine as a Inducer with Expression of Platelet-Derived Growth Factor α as a Maturity Marker

Background: The present study investigated the functional maturity of oligodendrocyte derived from rat bone marrow stromal cells (BMSC). Methods: The BMSC were isolated from female Sprague-Dawley rats and evaluated for different markers, such as fibronectin, CD106, CD90, Oct-4 and CD45. Transdifferentiation of OLC from BMSC was obtained by exposing the BMSC to DMSO and 1 µM all-trans-retinoic acid during the pre-induction stage and then induced by heregulin (HRG), platelet-derived growth factor AA (PDGFR-α), fibroblast growth factor and T3. The neuroprogenitor cells (NPC) were evaluated for nestin, neurofilament 68, neurofilament 160 and glial fibrillary acidic protein gene expression using immunocytochemistry. The OLC were assessed by immunocytochemistry for O4, oligo2, O1 and MBP marker and gene expression of PDGFR-α was examined by RT-PCR. Results: Our results showed that the fibronectin, CD106, CD90, CD45 and Oct-4 were expressed after the fourth passage. Also, the yield of OLC differentiation was about 71% when using the O1, O4 and oligo2 markers. Likewise, the expression of PDGFR-α in pre-oligodendrocytes was noticed, while MBP expression was detected in oligodendrocyte after 6 days of the induction. Conclusion: The conclusion of the study showed that BMSC can be induced to transdifferentiate into mature OLC. Key Words: Bone marrow stromal cell, Triiodothyronine, Platelet-derived growth factor α     

Quercetin attenuates H2O2‐induced toxicity of rooster semen during liquid storage at 4°C

Current study was carried out to examine the protective effects of quercetin against toxicity induced by hydrogen peroxide in rooster semen in vitro. Semen samples were collected from ten roosters (Ross 308 broiler breeder males, 32 weeks old) twice a week by abdominal massage method. Samples with ≥70% progressive motility were selected, pooled, diluted and used for the study. Experimental groups consisted of negative control, control that received solvent of quercetin, H₂O₂ (40 μM) and combination groups which incubated with constant dose of H₂O₂ (40 μM) plus various levels of quercetin (20, 40 and 80 μM). Measurement of total hydroperoxide (HPO), malondialdehyde (MDA), nitric oxide (NO), total antioxidant capacity (TAC) and superoxide dismutase activity as well as routine sperm tests were done at 0, 24 and 48 hr of storage at 4°C. Results revealed that exposure to hydrogen peroxide significantly increased HPO (138.43 ± 7.32 vs. 66.08 ± 3.97 μmol/g protein), MDA (7.21 ± 0.08 vs. 5.71 ± 2.16 μmol/g protein) and NO (0.367 ± 0.013 vs. 0.215 ± 0.011 μmol/g protein) levels and decreased sperm progressive motility (27.28 ± 1.21 vs. 47.49 ± 1.29%), and amounts of TAC (11.49 ± 0.39 vs. 15.70 ± 0.79 mmol/g protein) compared to control at 24 hr (p < 0.05). Changes at mentioned variables were repeated at 48 hr of storage. Also, co‐administration of quercetin (especially at 40 and 80 μM) with hydrogen peroxide restored the toxic effects of hydrogen peroxide on rooster semen parameters such as primary and secondary lipid peroxidative indicators and other evaluated variables. The study concluded that rooster semen enrichment with quercetin would protect lipid peroxidative and nitrosative hydrogen peroxide‐mediated damage during cold liquid storage of rooster semen.     

Differentiation of human umbilical cord mesenchymal stem cell into germ-like cell under effect of co-culture with testicular cell tissue

Supplements produced by mouse testicular cells (mTCs) and the interaction between cells can increase the differentiation rate of human umbilical cord mesenchymal stem cells (hUCMSCs) into the germ-like cells. We studied the differentiation rate of hUCMSCs into the germ-like cells under effect of mTCs co-culturing. Isolated hUCMSCs from postpartum human umbilical cords were cultured. Then, the expression of mesenchymal (CD73, CD90 and CD105) and haematopoietic (CD34 and CD45) markers of hUCMSCs were confirmed by flow cytometry. Then, the hUCMSCs were cultured in four distinct groups: (a) control, (b) co-culture until D0, (c) co-culture until D5 and (d) co-culture until D10, in order to differentiate into the germ-like cells. After 10 days, the expression of OCT4, VASA, Fragilis and SYCP3 genes were examined by Real-Time qPCR. The flow cytometry indicated a high expression of mesenchymal markers and a low expression of haematopoietic markers (CD73:98.6%, CD90: 99.1%, CD105: 99.5%, CD34: 4.22% and CD45: 2.54%). The expression of OCT4 decreased during the time while the expression of VASA, Fragilis and SYCP3 markers increased in the co-culture with testicular cells (p value <.05). Co-culture with mTCs may be used as an effective method to differentiate hUCMSCs into germ-like cells.     

Effect of dystocia on subsequent reproductive performance and functional longevity in Holstein cows

The objective of this study was to evaluate the effect of dystocia on the reproductive performance and functional longevity in Iranian Holsteins. Data consisted of 1 467 064 lactation records of 581 421 Holstein cows from 3083 herds which were collected by the Animal Breeding Center of Iran from April 1987 to February 2014. Reproduction traits in this study included interval from first to second calving, days open and days from first calving to first service. The generalized linear model was used for the statistical analysis of reproductive traits. Survival analysis was performed using the Weibull proportional hazards models to analyse the impact of dystocia on functional longevity. The incidence of dystocia had an adverse effect on the reproductive performance of dairy cows. Therefore, reproductive traits deteriorated along with increase in dystocia score (p < 0.05). The culling risk was increased along with increase in the score of dystocia (p < 0.0001). The greatest culling risk was observed in primiparous cows, small herds and low‐yielding cows (p < 0.0001). Also, the lowest culling risk was found for cows calving at the youngest age (<27 months), and cows with age at first calving >33 months had the greatest risk (p < 0.0001). The results of current study indicated that dystocia had important negative effects on the reproductive performance and functional longevity in dairy cows, and it should be avoided as much as possible to provide a good perspective in the scope of economic and animal welfare issues in dairy herds.     

The effects of onion and salt treatments on essential oil content and composition of Rosa damascena Mill

Due to the low oil content of Rosa damascena Mill., on one hand, and the wide spectrum of its application, on the other hand, it is desirable to find ways to increase the oil yield and also its quality. Therefore, in the present study the effects of chopped onion (1.0, 1.5, 2.0 and 4.0 g per 220 g of rose flower) and sodium chloride (11.0, 15.5, 22.0 and 30.0 g per 220 g of rose flower) which were added to rose flowers before distillation, were investigated on the quality and quantity of the oil. The quantity was measured as the ratio of oil to rose flower (w/w, %) and the quality was evaluated using the oil composition especially the content of monoterpene alcohols and hydrocarbons. Compared with corresponding control sample, treating with 1.5 g of chopped onion increased the oil content by 41% but it resulted in a lower quality; whereas in the presence of 1.0 g of onion, an increase of 30% and a decrease of 18% were observed in the percentage of monoterpene alcohols and hydrocarbons, respectively without any considerable change in the oil content. However, the effect of sodium chloride was somehow different from that of onion; such that in the optimal amount (22.0 g), the salt had no considerable effect on the quality but an increase of 21% in the oil yield.