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     

Effects of emulsified versus saline administration of GnRHa on induction of ovulation in rainbow trout, Oncorhynchus mykiss

The effects of saline-dissolved or Freund's incomplete adjuvant (FIA)-emulsified GnRHa treatment on the induction of ovulation in rainbow trout, Oncorhynchus mykiss, were examined. The FIA-emulsified GnRHa was first diluted in 0.25 ml physiological saline and then mixed with an equal volume of FIA. Fish were selected in the beginning of the spawning season and were allocated into four groups and were treated intraperitoneally with (a) 0.5 ml of emulsified GnRHa (GnRHa–FIA), (b) 0.5 ml saline-dissolved GnRHa in a single injection (GnRHa-1), (c) 0.5 ml saline-dissolved GnRHa in two injections spaced 1 d apart (GnRHa-2) and (d) 0.5 ml of saline (Control). The GnRHa dose in all hormone treatments was 25 µg kg^− 1. All fish in the FIA–GnRHa and GnRHa-2 groups ovulated within 10 and 11 d after treatment, respectively. In contrast, only 75% in the Control fish and 60% of the fish in the GnRHa-1 group ovulated within 36 d after treatment. None of the treatments caused any pre- or post-spawning mortality in the broodstock. Fertilization, eyeing and hatching percentages of the produced progeny were normal in all the treatment groups and did not differ significantly among them. In conclusion, FIA-emulsified GnRHa can be effective in advancing the onset of and synchronizing the ovulation of rainbow trout within a two-week period, thus shortening the egg collection period, without affecting broodstock survival and egg quality.     

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.     

Silicon Ameliorates the Adverse Effects of Salinity on Turfgrass Growth and Development

An experiment was conducted to assess the effect of foliar application of potassium silicate on Cynodon dactylon [L.] Pers., Festuca arundinacea Schreb. and Lolium perenne L. With increased salinity level, chlorophyll content and relative water content (RWC) in these three turfgrasses were reduced significantly when compared with the untreated control. Supplementary silicon (Si) ameliorated the adverse effects of salinity on chlorophyll content. Silicon treatments decreased proline at all salinity levels. Moreover, addition of Si increased shoot length and shoot number in all turfgrasses. Sodium (Na) concentration was increased in both leaves and roots of turfgrasses at high salinity level; however, Si treatment significantly reduced Na concentration in all of them. Silicon increased Potassium concentration in shoots and roots (though not significantly) in most cases. It can be concluded that Si alleviated the adverse effects of salt stress in all turfgrasses.     

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.     

Growth performance,muscle and liver composition,blood traits,digestibility and gut bacteria of beluga (Huso huso) juvenile fed different levels of soybean meal and lactic acid

The objective of this study was to assess the effects of fishmeal (FM) replacement with 0, 350 or 700 g/kg soybean meal (SBM) in combination with the supplementation of lactic acid (LA; 0, 10 or 20 g/kg) in the diets of juvenile beluga sturgeon (Huso huso; 700 ± 30 g). Nine isonitrogenous (400 g/kg protein) and isoenergetic (18 MJ/kg) diets were fed to beluga ad libitum, three times a day, for 60 days. The results showed that replacing FM with SBM without LA significantly reduced fish growth; on the other hand, LA supplementation had positive effects on fish fed diets that FM was replaced by SBM (p < .05). Increasing SBM in the diet altered the fatty acid profiles of the fish, reducing long‐chain polyunsaturated fatty acids and the n‐3/n‐6 fatty acids. High amounts of SBM (700 g/kg) caused reductions in the haematocrit, glucose and cholesterol levels in the blood (p < .05). In addition, the digestibility of protein, fat, dry matter and phosphorus was reduced when replacing FM with SBM, however, adding LA to the diets increased fish performance (p < .05), and this improvement was sharper in 2% LA groups. The number of LA bacteria increased significantly with the dietary supplementation of LA (p < .05). Based on these results, replacing 350 g/kg of FM with SBM and adding 20 g/kg LA to their feed do not negatively affect the biological and physiological indices of beluga.