Novel complex Re-Arrangement of ARG1 commonly shared by unrelated patients with Hyperargininemia

Background

Hyperargininemia is a very rare progressive neurometabolic disorder caused by deficiency of hepatic cytosolic arginase I, resulting from mutations in the ARG1 gene. Until now, some mutations were reported worldwide and none of them were of Southeast Asian origins. Furthermore, most reported mutations were point mutations and a few others deletions or insertions.

Objective

This study aims at identifying the disease-causing mutation in the ARG1 gene of Malaysian patients with hyperargininemia.

Methodology

We employed a series of PCR amplifications and direct sequencing in order to identify the mutation. We subsequently used quantitative real-time PCR to determine the copy number of the exons flanking the mutation. We blasted our sequencing data with that of the reference sequence in the NCBI in order to obtain positional insights of the mutation.

Results

We found a novel complex re-arrangement involving insertion, inversion and gross deletion of ARG1 (designated g.insIVS1 + 1899GTTTTATCAT;g.invIVS1 + 1933_ + 1953;g.delIVS1 + 1954_IVS2 + 914;c.del116_188;p.Pro20SerfsX4) commonly shared by 5 patients with hyperargininemia, each originating from different family. None of the affected families share known relationship with each other, although four of the five patients were known to have first-cousin consanguineous parents.

Conclusion

This is the first report of complex re-arrangement in the ARG1. Further analyses showing that the patients have shared the same geographic origin within the northeastern part of Malaysia prompted us to suggest a simple molecular screening of hyperargininemia within related ethnicities using a long-range PCR.     

三峡库区岸生植物野古草(Arundinella anomala Steud.)光合作用对水淹的响应

为了阐明水淹对三峡库区岸生植物野古草光合作用的影响,模拟了三峡库区消落带水淹发生情况,考察了在不同水淹处理下野古草(Arundinella anomalaSteud.)的光合及叶绿素荧光特性。实验设置了对照(不进行水淹,常规供水管理)、半淹(植株置于水中,植株地上部分一半被淹没)、水下0.5m(植株置于水中,植株顶部在水面下0.5m)、水下2m(植株置于水中,植株顶部在水面下2m)4个不同的水淹深度和02、0、40d和60d等4个不同的水淹时间处理,测定了在不同水淹深度和水淹时间处理下野古草的净光合速率、总叶绿素含量、PSⅡ的最大光化学效率、电子传递速率、表观量子效率、叶绿素利用效率与羧化效率。结果发现,在水淹前期,水淹对野古草的光合特性影响较小,直到水淹60d后,才对野古草的光合特性产生明显影响,且影响程度随水淹深度的不同而不同。野古草在水淹20d和水淹40d后,各水淹处理的净光合速率与对照相比无明显降低,其中水淹20d后,半淹处理的野古草叶片净光合速率比对照还高出16.1%。水淹60d后,水下0.5m和水下2m的净光合速率显著低于对照和半淹,其净光合速率分别为7.51μmol.m-2.s-1和9.15μmol.m-2.s-1。结果表明,水淹20d和40d对野古草的电子传递速率、表观量子效率和羧化效率没有影响。水淹处理60d后,与对照植株相比,半淹处理植株的电子传递速率、表观量子效率、叶绿素利用效率和羧化效率没有明显变化,但水下0.5m和水下2m处理植株的电子传递速率、表观量子效率、叶绿素利用效率和羧化效率有明显降低。在整个实验期间,半淹处理植株的净光合速率、电子传递速率、表观量子效率和羧化效率没有受到任何不利影响。尽管在水淹60d后水下0.5m和水下2m处理植株的净光合速率、电子传递速率、表观量子效率、叶绿素利用效率和羧化效率降低,但降低后的数值仍不低于甚至高于一些自然生长的未受水淹的植物物种。研究表明,野古草对水淹具有很好的耐受能力,是一种可以用于三峡库区消落区植被构建的优良植物物种。     

云南省恙螨区系及垂直分布研究

2000～2004年对云南省恙螨进行调查,发现云南省内恙螨192种,主要分布于中西部及中南部的热带及亚热带气候区,垂直高度多在1 000 m以下或1 500～2 500m范围.古北界种类有114种,占59%,东洋界种类较少为6%,35%的种类为跨界分布.中华纤恙螨leptotrombidium sinicum和小板纤恙螨L.scutellare为优势螨种.结合其他资料发现,该省广布种类10种,特有种5种,广宿主种及窄宿主种类均较多.少数螨种仅发现于3000 m以上的高海拔温带气候区或500m以下的低海拔热带气候区.恙螨分布与地域、海拔及气候因素有关.     

云南省恙螨名录初报

2000-2004年对云南省寄生恙螨进行现场调查,共采集到恙螨192种,结合其他文献资料报道,目前云南省境内已经发现的恙螨至少有222种,隶属3亚科27属。本文归纳总结了到目前为止云南省所记载恙螨的名称及所寄生宿主。     

云南省小兽体表革螨名录初报

1994年至2004年在云南省境内共调查了25个县（市）,共捕获53种啮齿目、食虫目、攀口目、兔形目和翼手目小兽10803只,从其体表分检出革螨68572只,隶属10科26属80种,其中云南新记录种12种;结合相关文献记载,到目前为止,整个云南省小兽体表革螨共有10科33属112种。     

Transgenic Rat Model of Neurodegeneration Caused by Mutation in the TDP Gene

TDP-43 proteinopathies have been observed in a wide range of neurodegenerative diseases. Mutations in the gene encoding TDP-43 (i.e., TDP) have been identified in amyotrophic lateral sclerosis (ALS) and in frontotemporal lobe degeneration associated with motor neuron disease. To study the consequences of TDP mutation in an intact system, we created transgenic rats expressing normal human TDP or a mutant form of human TDP with a M337V substitution. Overexpression of mutant, but not normal, TDP caused widespread neurodegeneration that predominantly affected the motor system. TDP mutation reproduced ALS phenotypes in transgenic rats, as seen by progressive degeneration of motor neurons and denervation atrophy of skeletal muscles. This robust rat model also recapitulated features of TDP-43 proteinopathies including the formation of TDP-43 inclusions, cytoplasmic localization of phosphorylated TDP-43, and fragmentation of TDP-43 protein. TDP transgenic rats will be useful for deciphering the mechanisms underlying TDP-43–related neurodegenerative diseases.     

Mutations in HPSE2 Cause Urofacial Syndrome

Urinary voiding dysfunction in childhood, manifesting as incontinence, dysuria, and urinary frequency, is a common condition. Urofacial syndrome (UFS) is a rare autosomal recessive disease characterized by facial grimacing when attempting to smile and failure of the urinary bladder to void completely despite a lack of anatomical bladder outflow obstruction or overt neurological damage. UFS individuals often have reflux of infected urine from the bladder to the upper renal tract, with a risk of kidney damage and renal failure. Whole-genome SNP mapping in one affected individual defined an autozygous region of 16 Mb on chromosome 10q23-q24, within which a 10 kb deletion encompassing exons 8 and 9 of HPSE2 was identified. Homozygous exonic deletions, nonsense mutations, and frameshift mutations in five further unrelated families confirmed HPSE2 as the causative gene for UFS. Mutations were not identified in four additional UFS patients, indicating genetic heterogeneity. We show that HPSE2 is expressed in the fetal and adult central nervous system, where it might be implicated in controlling facial expression and urinary voiding, and also in bladder smooth muscle, consistent with a role in renal tract morphology and function. Our findings have broader implications for understanding the genetic basis of lower renal tract malformations and voiding dysfunction.     

Somatic embryogenesis and vegetative cutting capacity are under distinct genetic control in Coffea canephora Pierre

The purpose of the study was to evaluate the possible genetic effect on vegetative propagation of Coffea canephora. Diversity for somatic embryogenesis (SE) ability was observed not only among two groups of C. canephora Pierre (Congolese and Guinean), but also within these different genetic groups. The results therefore showed that, under given experimental conditions, SE ability is depending on genotype. Furthermore the detection of quantitative trait loci (QTLs) controlling the SE and cutting abilities of C. canephora was performed on a large number of clones including accessions from a core collection, three parental clones and their segregating progenies. On the one hand we detected eight QTLs determining SE. Six positive QTLs for SE ability, whatever the criteria used to quantify this ability, were localized on one single chromosome region of the consensus genetic map. Two negative QTLs for SE ability (frequency of micro calli without somatic embryo) were detected on another linkage group. Deep analysis of the six QTLs detected for SE ability came to the conclusion that they can be assimilated to one single QTL explaining 8.6–12.2% of the observed variation. On the other hand, two QTLs for average length of roots and length of the longest sprouts of cuttings were detected in two linkage groups. These QTLs detected for cutting ability are explaining 12–27% of the observed variation. These observations led to conclude that SE and cutting abilities of C. canephora Pierre appeared to be genetic dependent but through independent mechanisms.     

MultiplexSSR: A pipeline for developing multiplex SSR‐PCR assays from resequencing data

Next‐generation sequencing has greatly promoted the investigation of single nucleotide polymorphisms, while studies of simple sequence repeats are sharply decreasing. However, simple sequence repeats still present some advantages in conservation genetics. In this study, an end‐to‐end pipeline referred to as MultiplexSSR was established to develop multiplex PCR assays in batches with highly polymorphic simple sequence repeats for capillary platforms from resequencing data. The distribution of single sequence repeats in the genome, the error profiles of genotypes and allelotypes, and the increase in the allele length range depending on the number of individuals were investigated. A total of 98% of single sequence repeats presented lengths of less than 100 bp. The error rate of the genotyping and allelotyping of dimeric patterns was ten times higher than those for other patterns. The error rate of allelotyping was less than that of genotyping. The allele length range reached approximate saturation with 10 individuals. This pipeline uses allele numbers to select highly polymorphic loci, masks loci with variation, and applies in silico PCR to improve primer specificity. The application of the developed multiplex SSR‐PCR assays validated the pipeline's robustness, showing higher polymorphism and stability for the developed simple sequence repeats and a lower cost for genotyping and providing low‐depth resequencing data from less than a dozen individuals for the development of markers. This pipeline fills the gap between next‐generation sequencing and multiplex SSR‐PCR.