Simultaneous Analysis of Multidrug Resistance 1(MDR1) C3435T,G2677T/A,and C1236T Genotypes in Hamadan City Population,West of Iran

Background: One of the limitations in the treatment of common diseases such as cancer chemotherapy is development of multidrug resistance (MDR). Polymorphisms could alter the expression level of MDR1 gene, which plays an important role in MDR. In this research, the frequency of C3435T, C1236T, and G2677T/A polymorphisms of MDR1 gene was investigated in a large group of population from Hamadan city to provide a sample data resource. Methods: Peripheral blood (2 ml) was taken, and DNA extraction was carried out. Multiplexed mutagenically separated PCR, which was followed by polyacrylamide gel electrophoresis and silver staining, was applied to detect the mentioned polymorphisms in 935 individuals. Sequencing performed for confirmation of gel electrophoresis resulted in 10 random cases. In total, alleles and genotypes of 933 persons (776 women and 157 men) were determined. Results: The most frequent alleles of the polymorphisms were: 3435T, C1236, and G2677. The most frequent genotypes were: 3435C/T, 1236C/T, and 2677G/A, and their concurrent presence was also found as the most frequent simultaneous genotypes. There was not any meaningful difference among the prevalence of these genotypes in groups of men and women. Conclusion: Our results were close to those of other studies performed in Iran and compared to the other ethnic groups, which showed more similarity to Asian peoples than Europeans. As an aspect of personalized medicine, it could be used by chemotherapists to improve the routine methods of cancer treatment.Key Words: Gene polymorphism, Multi-drug resistance, Neoplasm     

Association between aflatoxin contamination and drought tolerance traits in peanut

The current study investigates the association between drought tolerance traits and aflatoxin contamination in peanut grown under long-term drought. Two field experiments were conducted at Khon Kaen University, Thailand using a split–split plot design with three drought stress levels as main plots, 11 genotypes as sub-plots, and two soil inoculations of Aspergillus flavus treatments as sub-sub-plots. The effects of temperature, soil moisture and A. flavus population on kernel colonization and aflatoxin contamination, and drought tolerance traits viz. specific leaf area (SLA) and root length density (RLD) were measured. The results demonstrated that elevated soil temperatures and reduced soil moisture, favored aflatoxin production. Drought in combination with higher levels of A. flavus inoculum load in the soil resulted in an increase in the fungal populations in the soil which in turn resulted in increased kernel colonization and subsequent aflatoxin contamination. A combination of SLA and RLD, and kernel colonization had a significant influence on aflatoxin contamination under drought conditions in both seasons (r = 0.73** and 0.76**). The results revealed that drought tolerance traits (SLA and RLD) could be contributing to resistance to aflatoxin contamination suggesting that a combination of SLA, RLD and kernel colonization could be used as selection criteria in selecting parents for aflatoxin resistance.     

大麦蛋白质含量与SSR标记的关联分析

为检测与大麦蛋白质含量相关联的SSR标记,对93份不同来源的大麦材料进行蛋白质含量测定和93个SSR标记分析,利用GLM和MLM模型进行SSR标记与蛋白质含量的关联分析。结果显示,93份材料共检测出406个等位变异,每个标记的等位变异数平均为4.4个,基因多样性和PIC变幅分别为0.021 3～0.862 0和0.021 0～0.648 5。通过群体结构分析将93份材料划分为5个亚群。GLM和MLM模型分析结果表明,在两种模型中均检测到9个SSR标记与大麦籽粒蛋白质含量相关联,且所检测到的标记相一致,对表现变异的解释率分别为7.49%～14.13%和7.30%～13.97%;两种模型中,均只有标记GMS061与蛋白质含量关联达到极显著水平(P0.01)。本研究结果可为后期标记辅助选择育种提供一定的理论依据。     

木薯栽培品种农艺性状与分子标记的关联分析

为寻找与木薯茎叶鲜重、块根鲜重、株高、开花、块根数和叶片叶绿素含量紧密关联的分子标记,利用145对SRAP,132对EST-SSR引物对19个木薯栽培品种进行多位点的扫描分析,并在分析群体的连锁不平衡水平基础上,通过关联分析软件TASSEL2.1将标记与这些栽培品种的以上农艺性状进行关联。结果表明:群体中非共线性的SRAP和EST-SS位点组合存在一定的连锁不平衡,在147个SRAP位点的1471个位点组合中支持的不平衡成对位点仅在木薯粗略基因组中占位点组合的0.815%(p0.01),132个ESTSSR位点的993种位点组合仅在木薯基因组中占位点组合的0.302%(p0.01);利用EST-SSR数据分析栽培品种的群体结构可划分为3个亚群,亚群的划分与群体亲缘类型相关联;共检测到29个与性状相关联的位点,其中与茎叶鲜重相关的位点11个,与块根鲜重相关的位点3个,与株高相关的位点3个,与是否开花相关的位点8个,与块根数相关的位点5个,与叶片叶绿素含量相关的位点2个。下一步工作是进行优等位基因的挖掘,找到与茎叶鲜重和株高相关的优异位点、等位变异及携带优异等位变异的载体材料。     

Differential expression of rheumatoid factor-associated crossreactive idiotypes in Iranian seropositive and seronegative patients with rheumatoid arthritis

INTRODUCTION: High levels of rheumatoid factors (RF) are detectable in serum of the majority of patients with rheumatoid arthritis (RA), but 5-10% of patients remain seronegative (SN). Despite clinical and genetic similarities between these two subsets of RA, it has been proposed that they may be regarded as distinct clinical entities. METHODS: In the present study a panel of monoclonal antibodies (mAb) recognizing RF-associated cross-reactive idiotypes (CRI) linked to the VH1 (G8), VH4 (LC1), VK3b (17-109) and a mAb recognizing the VK3 subgroup (C7) of immunoglobulin variable region (IgV) gene products were used to quantitate the level of expression of these gene products in serum and synovial fluid of 35 seropositive (SP) and 8 SN RA patients by capture ELISA. RESULTS: While the concentration and relative proportion of the IgV are recognized by the mAb G8, 17-109 and C7 were significantly higher in serum and synovial fluid of the SP RA, compared to the SN-RA patients (G8, p = 0.009; 17-109, p = 0.0001; C7, p = 0.001). The CRI recognized by the mAb LC1 was highly represented in serum and synovial fluid of the SN-RA patients. There have been no significant differences in the level of expression of these IgV gene products (other than the product recognized by C7 mAb in SP patients) between serum and synovial fluid of either group of patients. CONCLUSION: Our results suggest that the expressed repertoire of Ig VH and VK genes in these two subsets of RA is differentially regulated and may be influenced by selective mechanisms leading to positive or negative selection of certain genes.     

Association of Lecithin Cholesterol Acyltransferase rs5923 Polymorphism in Iranian Individuals with Extremely Low High-Density Lipoprotein Cholesterol: Tehran Lipid and Glucose Study

Background:

The serum concentration of high-density lipoprotein cholesterol (HDL-C) is one of the important heritable risk factors for cardiovascular disease and is a target for therapeutic intervention. In this study, we aimed to evaluate the effects of lecithin cholesterol acyltransferase (LCAT) gene polymorphism rs5923 on LCAT enzyme activity and serum HDL-C concentration.

Methods:

The study population was selected from consecutive individuals with HDL-C ≤ 5^th percentile (n = 73) and extremely high HDL-C ≥ 95^th percentile (n = 57) who had participated in the Tehran Lipid and Glucose Study. The rs5923 polymorphism was genotyped using direct sequencing. LCAT activity was measured by fluorometric assay kit, and lipid concentrations were measured using the enzymatic colorimetric method.

Results:

The genotype frequencies were significantly different between the high HDL-C group (CC 94.7%, CT 5.3%) and the low HDL-C group (CC 83.6%, CT 16.4%) (P = 0.048). The T-allele frequencies in subjects with low and high HDL-C were 0.082 and 0.026, respectively (P = 0.16). The association of the single-nucleotide polymorphism rs5923 with low HDL-C was not statistically significant after adjustment for age, sex, and BMI (odd ratio = 2.65, 95% confidence interval = 0.32-21.5, P = 0.36, regression logistic analysis). Also, the effects of LCAT enzyme activity did not depend on the HDL-C level (P = 0.24).

Conclusion:

rs5923 polymorphism is not associated with low HDL-C levels in Iranian population. Key Words: Polymorphism, Single nucleotide, Lipoproteins     

Clinical and Genetic Characteristics of Splicing Variant in CYP27A1 in an Iranian Family with Cerebrotendinous xanthomatosis

Background:CTX is a rare congenital lipid-storage disorder, leading to a progressive multisystem disease. CTX with autosomal recessive inheritance is caused by a defect in the CYP27A1 gene. Chronic diarrhea, tendon xanthomas, neurologic impairment, and bilateral cataracts are common symptoms of the disease. Methods:Three affected siblings with an initial diagnosis of non-syndromic intellectual disability were recruited for further molecular investigations. To identify the possible genetic cause(s), WES was performed on the proband. Sanger sequencing was applied to confirm the final variant. The clinical and molecular genetic features of the three siblings from the new CTX family and other patients with the same mutations, as previously reported, were analyzed. The CYP27A1 gene was also studied for the number of pathogenic variants and their location. Results:We found a homozygous splicing mutation, {"type":"entrez-nucleotide","attrs":{"text":"NM_000784","term_id":"1519312912"}}NM_000784: exon6: c.1184+1G>A, in CYP27A1 gene, which was confirmed by Sanger sequencing. Among the detected pathogenic variants, the splice site mutation had the highest prevalence, and the mutations were mostly found in exon 4. Conclusion:This study is the first to report the c.1184+1G>A mutation in Iran. Our findings highlight the other feature of the disease, which is the lack of relationship between phenotype and genotype. Due to nonspecific symptoms and delay in diagnosis, CYP27A1 genetic analysis should be the definitive method for CTX diagnosis. Key Words: Cerebrotendinous xanthomatosis, CYP27A1, Intellectual disability, Iran, Whole exome sequencing     

咖啡小粒种六倍体和中小粒杂种四倍体胚乳的组织培养

进行了利用咖啡胚乳离体培养技术培育六倍体咖啡胚乳植株及四倍体中小粒种咖啡种间杂种的研究。结果表明:将小粒种咖啡自花授粉所得六倍性胚乳及以中粒种咖啡为母本、小粒种咖啡为父本进行杂交所获得的四倍性种间杂种胚乳,分别在附加有6-BA1~2mg/L和NAA2mg/L的改良MS培养基上培养,能诱导出有效的愈伤组织;在附加有6-BA或KT0.5mg/L,NAA0.1mg/L的改良MS培养基上,愈伤组织能分化出各种类型的胚状体;在附加有NAA或IBA0.5mg/L及2g/L活性炭的1/2MS培养基上,胚状体可被诱导生根而再生完整的六倍体咖啡胚乳植株和四倍体种间杂种咖啡胚乳植株。      

Role of endogenous vitamin E in renal ischemic preconditioning process: differences between male and female rats

Background: Antioxidants such as α-tocopherol (vitamin E) and β-carotene (vitamin A) play an important role in protective effect of repeated brief periods of ischemia, namely ischemic preconditioning (IPC). Values of these antioxidants were investigated and compared after induction of ischemia reperfusion (IR) and kidney IPC in both male and female rats. Methods: Forty eight Wistar rats were divided randomly into six groups of 8: groups A and B (male and female controls, respectively), group C (male IR or IR cases), group D (female IR cases) and groups E and F (male and female IPC cases, respectively). In groups C and D, ischemia was induced by clamping of left renal arteries for 45 min. In groups E and F, rats underwent four cycles of 4 min of arterial clamping and 11 min of de-clamping before final 45 min ischemia induction. Afterward, serum was collected to assess the blood urea nitrogen, creatinine and vitamins E and A values. Renal tissues were obtained for histological assessments. Results: α-tocopherol levels in male and female rats showed a significant increase in IPC compared with IR group (P<0.01) and also in female IPC compared with male IPC group. β-carotene levels had no significant variations. Histological evaluation showed that IR-induced renal injuries were less in female rats. Also, protective effects of IPC were more in female rats (P<0.01). Conclusions: Renal IPC reduced damages in both male and female rats, but tissue injuries in females were decreased much more along with the increase of endogenous vitamin E. Key Words: α-tocopherol, β-carotene, Antioxidant, Reperfusion, Ischemic preconditioning (IPC)     

A2、A3类型高粱F2代花粉碘染色率与田间自交结实率的关系

以高粱3个A2类型杂交种（A2TX622×晋粱5号、A2V4×1383-2、A2V4×晋粱5号）和3个A3类型杂交种（A3299×1174、A32457×1174、A3JW×1174）的F2群体为材料,来进行A2、A3类型高粱F2代花粉碘染色率与田间自交结实率之间关系的研究。结果表明：3个A2类型F2群体联合分析结果的回归方程为RT=1.188×R0.25＋0.906×R0.75＋0.915×R1.0,3个A3类型F2群体联合分析的回归方程为RT=0.188×R0.25＋0.832×R0.75＋0.933×R1.0,所有6个F2群体联合分析的回归方程为RT=0.193×R0.25＋0.896×R0.75＋0.949×R1.0。RT与R0.25、R0.75和R1.0之间确实存在显著或极显著的回归关系,而且品种间存在差异。     

Association between climate indices,aridity index,and rainfed crop yield in northeast of Iran

Agricultural drought occurs when there is a deficit in soil water supply to crops. Severe drought limits crop water availability and reduces yield. Rainfed crop production is very vulnerable to drought conditions and farmers in northeast of Iran who heavily depend on their rainfed cereals production usually suffer from drought occurrence. Based on history, any severe drought resulted in severe financial problems and forced the affected farmers to move to cities in search of alternative jobs. Any possibility to enable the farmers to mitigate or adapt to drought is highly required. In this study, the relationship between aridity index (AI) and detrended crop yield (1985–2005) of selected crops (wheat and barley) and the influence of three climate indices (AO, NAO and NINO-3.4) were assessed for Khorasan province in northeast of Iran. All associations were assessed at annual, seasonal (wet and dry seasons) and monthly scale considering both concurrent and lag correlations (1-year and 2-year lag). Our results indicated a significant correlation (P < 0.05) between the AI and crops yield mostly in central Khorasan province. Our study also showed that correlation coefficient between AI and barley yield was stronger than AI and wheat yield across all study locations. Seasonal (wet) AI showed significant correlation with crops yield. These results demonstrated that, in some areas of Khorasan, drought is one of the key causes of interannual yield variability. We also observed a significant association between NAO and NINO-3.4 with AI. Precipitation is one of the components of AI, so AI response to NAO and NINO-3.4 can be related to the observed association between this index and precipitation. It seems that these indices could be useful tools to monitor drought patterns and subsequent yield variability in some regions of Khorasan province.     

Correlation between catecholamine levels and outcome in patients with severe head trauma

Some studies have shown that catecholamines and the changes in their levels during and after head trauma can be useful in predicting the outcome in head trauma patients. The goal of this study is to search for a probable relation between urine levels of catecholamines and prognosis in patients with severe head trauma. Fifty four patients with severe head trauma Glasgow Coma Scale (GCS < or = 8) on admission time were recruited in Imam Reza Hospital within one. These patients were included when having no major accompanying trauma in other organs. Twenty four hour urine was collected after admission and levels of metanephrine and nor-metanephrine were measured. The relation between urine levels of these metabolites with final outcome and also with GCS at admission, 24, 48 h and 1 week after admission and discharge time and Glasgow Outcome Scale (GOS) were studied. Fifty two patients, 48 males and 4 females with a mean age of 32.3 +/- 14.7 (3-72) years were included. The main underlying etiologies were motorcycle (46.2%) and car accidents (25%). Diffuse axonal injury, brain contusion and subdural hematoma were three main diagnoses (28.8, 17.3 and 15.4% of the cases, respectively). 19 (36.5%) of the patients expired within the study period. The mean level of metanephrine and normetanephrine in urine were 207.9 +/- 200.5 and 330.2 +/- 218.4 microg in 24 h, respectively. There was no meaningful relation between urine levels of these metabolites and any of GCS and GOS. There was also no meaningful relation between these parameters and final prognosis in patients.     

Close Association between Aleurone Traits and Lipid Contents of Rice Grains Observed in Widely Different GeneticResources of Oryza sativa

Abstract

We investigated 321 varieties to identify the aleurone traits that are closely related to the lipid content of rice. Brown rice seeds were cut crosswise near the center with a razor blade and the cut surface was stained with Oil Red O, and then observed under integrated fluorescence microscope (BZ9000). We found wide variations among varieties in the area stained, but the japonica group contained many varieties with a large stained area. We selected 17 representative varieties covering the whole range of stained areas and confirmed the thickness of aleurone layer using Kawamoto’s film method (micro-thin sections that were created with cryomicrotome). Aleurone traits were examined under a light microscope and measured. We found a strong correlation between the area of the aleurone layer and the stained area (r = 0.799***) and between the thickness of the aleurone layer and the thickness of the stained region (r = 0.543*). The area of the stained region therefore provides a fast and effective indicator for selecting varieties for the thickness of the aleurone layer. We also measured the amount (mg per 100 seeds) and proportion of triacylglycerols (TAGs) to 1 g of seed (mg g-¹) in brown rice seeds without embryos. The amount and proportion of TAG to 1 g of seed were both significantly correlated with the area, average thickness and percentage of aleurone layer. This suggests that the aleurone traits will be good indicators for the selection of varieties with high levels of TAG.     

Association between root growth angle and root length density of a near-isogenic line of IR64 rice with DEEPER ROOTING 1 under different levels of soil compaction

DEEPER ROOTING 1 (DRO1) of rice controls the gravitropic response of root growth angle. In order to clarify the effects of DRO1 on root growth angle and root length density under different soil resistance to penetration, and to quantify the relationship between root growth angle and root length density, we assessed the root growth of Dro1-NIL (a near-isogenic line homozygous for the Kinandang Patong allele of DRO1 in the IR64 background) under upland Andosol field conditions in Japan in 2013 and 2014. The trial included three levels of soil compaction (none, moderate, and hard). Root length density at a depth of 30 to 60 cm was largest in Kinandang Patong, followed by Dro1-NIL, and was least in IR64 in both years and in all compaction treatments. Root length density at this depth decreased with hard compaction (to 70% of control) and increased with moderate compaction (to 135%). The number of roots with a deep angle (i.e. 45° to 90° from the horizontal) measured by the basket method was similar at maximum tillering and maturity stages, and its value as a proportion of the total number of roots was strongly correlated with the root length density at 30 to 60 cm in both years, which demonstrates the importance of a deep root angle for the development of deep roots. Dro1-NIL had a higher proportion of deep roots than IR64, but the difference was small under hard compaction, with a significant genotype × compaction interaction.     

抗虫抗除草剂转基因水稻B1C893的获得与鉴定

利用农杆菌介导法将Cry1Ca#和Bar基因转化水稻恢复系R893,获得6株再生植株。经过分子鉴定,发现Bar基因和Cry1Ca#基因成功整合到水稻基因组并稳定表达。测定其中单拷贝且可育的转基因株系苗期不同器官中Cry1Ca蛋白的表达量,发现根系中最低而叶片中最高,其中最高的第6号株系叶片中含量达到35.07μg/g。获得的转基因株系对除草剂草铵膦和稻纵卷叶螟均具有良好的抗性。     

旱地保护性耕作条件下不同冬小麦品种部分生理指标与 WUE的关系

为给旱地保护性耕作条件下小麦高水分利用效率品种的筛选以及高产节水栽培提供依据,以烟Blu 6439、洛旱6号、石家庄8号、石麦15和烟农21为供试材料,研究了小麦不同生育期的LAI、植被归一化指数(NDVI)、SPAD值、净光合速率(Pn)与水分利用效率(WUE)的关系.结果表明,旱地保护性耕作条件下,不同基因型小麦的LAI和NDVI在抽穗期和灌浆中期与WUE存在抛物线关系.旗叶SPAD值和Pn与WUE呈直线关系.说明在旱地保护性耕作条件下,SPAD值和Pn较高的小麦品种具有高效用水潜能,可作为筛选高产节水品种的重要指标.     

小麦Glu-A1位点1和2*亚基近等基因系间品质差异的研究

为了解小麦G lu-A 1位点的1亚基和2*亚基间的遗传差异,2000~2002年对1和2*亚基的2对近等基因系(N IL s,分别来自龙麦19和克丰6号)进行了分析。结果表明,1亚基与其相应的2*亚基近等基因系间在蛋白质含量、湿面筋/干面筋比值、沉降值/干面筋比值的平均值分别为15.3%/15.3%、2.65/2.71、2.33/2.28;吸水率、形成时间、稳定时间、软化度、断裂时间、评价值的平均值分别为63.0%/63.0%、4.1 m in/4.3 m in、9.4 m in/9.1 m in、63.3 FU/59.2 FU、13.9 m in/12.7 m in、58.7/59.5;最大阻力、延伸性和拉伸面积的平均值分别为270 EU/256 EU、18.5 cm/18.9 cm、71.8 cm2/65.1 cm2。这些品质性状间的差异在统计学上均不显著。最后还讨论了两种亚基在小麦品质育种中的应用。     

香蕉枯萎病菌内源报告基因细胞周期蛋白C1基因FocFCC1的鉴定及分析

本研究克隆鉴定了香蕉枯萎病菌4号生理小种（Foc4）的细胞周期蛋白C1（Fusarium Cyclin C1,FCC1）基因FocFCC1,采用Split-marker同源重组技术获得基因敲除突变体ΔFocFCC1,通过比较突变体和野生菌株在生长速度、产孢量和致病力等方面的差异,探索了FocFCC1基因缺失对香蕉枯萎病菌生物学功能的影响,并评估了FocFCC1作为枯萎菌内源报告基因的可行性。结果表明：与Foc4野生型菌株相比,ΔFocFCC1菌丝生长缓慢、菌丝畸形及产孢量减少,对巴西蕉苗的致病力明显减弱,推测FocFCC1基因在Foc4的生长发育、产孢及致病力等方面具有重要作用。以FocFCC1为靶标基因,评估了两种基因敲除重组方法介导的基因敲除效率,利用红色表型作为判断依据,挑取再生板上有红色表型的转化子进行PCR检测,Split-marker重组方法获得阳性转化子比例为91.7%,而多片段装配融合方法获得阳性转化子的比例为64%。ΔFocFCC1出现典型红色菌落,红色表型与PCR检测的FocFCC1阳性敲除转化子一一对应,FocFCC1基因敲除后红色表型肉眼容易分辨,可作为香蕉枯萎病菌内源报告基因探索新的遗传操作技术在该菌上应用的可行性。     

陆地棉杂交种F1株内变异与产量性状的关系研究

 试验对陆地棉F₁铃重、绒长、衣分、子指的株内横向变异量和纵向变异量与构成皮棉产量的4个主要性状进行了分析,并重点对铃重、衣分的横向变异量和纵向变异量与直接构成皮棉产量的4个性状进行了典型相关分析。通过典型变量的筛选,第一对典型变量达到极显著差异,其中铃重的横向变异与单株皮棉产量均具有较大的信息载荷量,但二者方向相反,说明铃重横向变异量大的类型不利于产量的提高。铃重横向变异量应尽可能为小值。