Significance of MEFV gene R202Q polymorphism in Turkish familial Mediterranean fever patients

Objective

Familial Mediterranean fever (FMF) is an autosomal recessive disorder characterized by recurrent attacks of fever and inflammation in the peritoneum, synovium, or pleura, accompanied by pain. The disease is associated with mutations in the Mediterranean fever (MEFV) gene, which encodes for the pyrin protein. The aim of this study was to explore the frequency and clinical significance of the R202Q (c.605G>A) polymorphism in exon 2 of the MEFV gene in a cohort of Turkish patients with FMF.

Methods

The study included 191 patients with FMF and 150 healthy controls. Genomic DNA was isolated and genotyped using polymerase chain reaction (PCR)-based restriction fragment length polymorphism (RFLP) assay for the MEFV gene R202Qpolymorphism.

Results

The genotype and allele frequencies of R202Q polymorphism showed a statistically significant difference between FMF patients and controls (p < 0.0001 and p = 0.0004, respectively) and especially the homozygous AA genotype was significantly higher in FMF patients than healthy controls (p = 0.0002; odds ratio = 6.27; 95% CI = 2.1–18.3). However no significant association was observed between clinical and demographic features of FMF patients and R202Qpolymorphism.

Conclusion

The results of this study showed that there was a high association between MEFV gene R202Q polymorphism and FMF. R202Q polymorphism should be included in routine molecular diagnosis of FMF patients.     

Frequency of MEFV gene mutations in Hatay province,Mediterranean region of Turkey and report of a novel missense mutation (I247V)

In the present study, 1000 patients with clinical suspicion of FMF were retrospectively reviewed to determine the spectrum of MEFV gene mutations by using DNA sequence analysis between September, 2008 and April, 2012. Sixteen different mutations and 55 different genotypes were detected in 618 of 1000 patients. Among 16 different mutations, R202Q (21.35%) was the most frequently observed mutation; followed by E148Q (8.85%), M694V (7.95%), M680I (2.40%), V726A (1.85%), M694I (0.95%), A744S (0.80%), R761H (0.55%), P283L (0.35%), K695R (0.20%), E230K (0.15%), L110P (0.10%), I247V (0.05%), G196W (0.05%) and G304R (0.05%). In the present study, a novel missense mutation (I247V) and a silent variant (G150G) were identified in the MEFV gene. On the other hand, P238L, G632A and G304R mutations are the first cases reported from Turkey. Our results indicated that MEFV mutations are highly heterogeneous in our study population as in other regions of Turkey and mutation screening techniques such as PCR-RFLP, amplification refractory mutation system or reverse hybridization do not adequately detect uncommon or novel mutations. Therefore, it was proven that sequence analysis of the MEFV gene could be useful for detection of rare or unknown mutations.     

Mutational analysis of the PRYSPRY domain of pyrin and implications for familial mediterranean fever (FMF)

Familial Mediterranean fever (FMF) is an autosomal, recessively inherited disease, characterized by recurrent fever and serositis that affects mainly patients of the Mediterranean basin. The gene responsible for FMF, named MEFV, was cloned and several missense mutations were found to be responsible for the disease. Based on a recent molecular analysis of MEFV gene mutations in 43 patients from Crete aiming to correlate specific genotypes and clinical manifestations of FMF, we were prompted to construct a three-dimensional model (3-D model) of the PRYSPRY domain of pyrin. The majority of the known MEFV mutations located on this domain have been classified, according to disease severity, and localized on this 3-D model. The functional consequences of these mutations and their implications on disease severity are discussed. Moreover, we report a putative novel missense mutation, S702C, which we identified in exon 10 of the MEFV gene and localized on the constructed 3-D model.     

The role of the pro-apoptotic protein Siva in the pathogenesis of Familial Mediterranean fever: A structural and functional analysis

Familial Mediterranean fever (FMF) is an autosomal, recessive disease, attributed to mutations in MEFV gene encoding pyrin, which is characterized by recurrent, acute and self-limiting attacks of fever as well as an increased neutrophil and monocyte apoptosis. Most disease-associated mutations in MEFV gene reside on the C-terminal PRYSPRY (B30.2) domain of pyrin, an area found to interact with the pro-apoptotic protein Siva. Because apoptotic events may be contributing to endogenous inflammation we hypothesized that mutations in pyrin may affect Siva-mediated apoptosis. The confirmation of this hypothesis would be of a great biological significance since it would be demonstrated a connection between apoptosis and inflammation. We used homology modeling to construct a 3-D model of Siva protein and the constructed model of Siva defined structural elements with potential of binding other proteins to induce apoptosis. Given that Siva protein binds pyrin as shown by transfection and immunoprecipitation experiments, apoptosis was assessed by FACS and Western blotting. No differences in rates of apoptosis in myeloid cells (THP-1) upon transfection with either wt pyrin or mutant forms of pyrin were found. Patients with FMF did not display any mutations in the Siva-1 (full length) gene. Siva-1 was not linked to pyrin in the major predicted FMF gene network constructed using a literature-curated gene signature for FMF. These results suggest that Siva-mediated unprovoked apoptosis is not likely to be involved in the pathogenesis of FMF.     

Localization of the familial Mediterranean fever gene (FMF) to a 250-kb interval in non-Ashkenazi Jewish founder haplotypes. The French FMF Consortium.

Chromosome 16p13.3 harbors a gene (MEF) associated with familial Mediterranean fever (FMF), a recessive disease very common in populations of Mediterranean ancestry. In the course of positional cloning of MEF, we genotyped 26 non-Ashkenazi Jewish FMF pedigrees (310 meioses) with 15 microsatellite markers, most of which were recently developed by Généthon. Identification of recombination events in the haplotypes allowed narrowing of the MEF interval to a region between D16S3124 (telomeric) and D16S475 (centromeric). Two markers, D16S3070 and D16S3275, a microsatellite marker isolated from a YAC that also contains D16S3070, showed no recombination with the disease. Linkage disequilibrium and haplotype analysis highlighted the existence of a founder haplotype in our population. The core ancestral alleles were present in 71% of MEF-bearing chromosomes at loci D16S3070 and D16S3275. Furthermore, identification of historical crossing-over events in these pedigrees indicated that MEF is located between these two loci, which are both contained in a 250-kb genomic fragment.     

Occurrence of epibenthic dinoflagellates in relation to biotic substrates and to environmental factors in Southern Mediterranean (Bizerte Bay and Lagoon,Tunisia): An emphasis on the harmful Ostreopsis spp., Prorocentrum lima and Coolia monotis

Harmful events associated with epibenthic dinoflagellates, have been reported more frequently over the last decades. Occurrence of potentially toxic benthic dinoflagellates, on the leaves of two magnoliophytes (Cymodocea nodosa and Zostera noltei) and thalli of the macroalgae (Ulva rigida), was monitored over one year (From May 2015 to April 2016) in the Bizerte Bay and Lagoon (North of Tunisia, Southern Mediterranean Sea). The investigated lagoon is known to be highly anthropized. This is the first report on the seasonal distribution of epibenthic dinoflagellates hosted by natural substrates, from two contrasted, adjacent coastal Mediterranean ecosystems. The environmental factors promoting the development of the harmful epibenthic dinoflagellates Ostreopsis spp., Prorocentrum lima and Coolia monotis were investigated. The highest cell densities were reached by Ostreopsis spp. (1.9 × 10³ cells g⁻¹ FW, in October 2015), P. lima (1.6 × 10³ cells g⁻¹ FW, in June 2015) and C. monotis (1.1 × 10³ cells g⁻¹ FW, in May 2015). C. nodosa and Z. noltei were the most favorable host macrophytes for C. monotis (in station L2) and Ostreopsis spp. (in station L3), respectively. Positive correlations were recorded between Ostreopsis spp. and temperature. Densities of the epibenthic dinoflagellates varied according to the collection site, and a great disparity was observed between the Bay and the Lagoon. Maximum concentrations were recorded on C. nodosa leaves from the Bizerte Bay, while low epiphytic cell abundances were associated with macrophytes sampled from the Bizerte Lagoon. The observed differences in dinoflagellate abundances between the two ecosystems (Bay-Lagoon) seemed not related to the nutrients, but rather to the poor environmental conditions in the lagoon.     

Biscogniauxia mediterranea associated with cork oak (Quercus suber) in Tunisia: Relationships between phenotypic variation,genetic diversity and ecological factors

Biscogniauxia mediterranea is a xylariaceous ascomycete responsible for ‘charcoal disease’, the most frequent disease of cork oak in the Mediterranean basin. However, little is known regarding this fungus in North Africa. This is the first report on the morphological and genetic diversity of B. mediterranea populations in Tunisia.A total of forty-eight isolates from cork oak trees, corresponding to three different sites and three tree parts, were identified as B. mediterranea based on morphological and molecular diagnosis. Variability among the isolates was evaluated using morphological traits and microsatellite-primed PCR (MSP-PCR). B. mediterranea isolates showed high variability in culture, specifically in colony color. A high level of genetic diversity within populations and within tree parts was detected (H = 0.417, I = 0.605, and H = 0.415, I = 0.601 respectively). Significant genetic differentiation among populations (G_ST = 0.116 and Φ_ST = 0.137; p < 0.001) was determined. However, a low genetic differentiation among tree parts (G_ST = 0.066 and Φ_ST = 0.018; p = 0.07) was observed. Principal coordinate analysis showed that genetic divergence was partially related to the geographical origin. Cluster analysis based on morphological traits showed that the grouping of the isolates occurs independently of the geographic location and the tree part. The effective number of alleles (Ne), the Nei's gene diversity index (H), the Shannon's information index (I), the colony color and the presence of exudates were significantly correlated with the altitude and the rainfall. The number of alleles (Na), the percentage of polymorphic loci (PPL), the texture and the density of the mycelium were significantly correlated with the temperature. These results provide a deeper understanding of population genetics of B. mediterranea and its adaptation to environmental conditions which could help to develop control strategies against charcoal canker disease.     

马铁菊头蝠(Rhinolophus ferrumequinum)回声定位声波与生境类型和环境因子的关系

2007年在吉林省罗通山自然保护区,利用超声波探测仪（Avisoft-SASLAB PRO）录制并分析不同生境中马铁菊头蝠的回声定位声波。结果显示马铁菊头蝠在不同类型生境中活动;各生境中回声定位声波参数存在显著差异（one-way ANOVA,P〈0.05）。从环境因子中通过主成分分析筛选出与其回声定位声波相关的植被、气候和地形因子,探讨回声定位声波与这些因子的相关性。结果显示FM1和FM2带宽与乔木高（r=-0.948,-0.825;P〈0.05）、FM1起始频率和FM2终止频率与林冠面积（r=-0.967,-0.958;P〈0.05）、FM1起始频率、FM2终止频率和峰频与湿度（r=-0.776、-0.875和-0.794,P〈0.05）、脉冲持续时间和脉冲间隔与平均灌木高均呈显著负相关（r=-0.911,-0.990;P〈0.05）,峰频与植被株数（r=0.756,P〈0.05）、脉冲持续时间与冠下高呈显著正相关（r=0.870,P〈0.05）。表明各种环境因子（植被因子、气候因子和地形因子）都在一定程度上影响回声定位声波,回声定位声波具有表型可塑性和生境适应性,这些特性决定了马铁菊头蝠生境利用的程度和可利用的资源。     

Effects of environmental and temporal factors on Glomeromycotina spores in sand dunes along the Gulf of Valencia (Spain)

AMF symbiosis in sand dunes is the key for maintenance of stable vegetation. The main goal of this work was to determine the effects of environmental and temporal factors on AMF living in sand dunes (Gulf of Valencia, Spain). Soil samples were collected seasonally at 6 sites, during 2 yrs, from three habitats and four plant species and the frequency and relative abundance of AMF was examined. AMF were more frequent in mobile than in embryonic dunes, in spring and in sites with old vegetation. Ten AMF species were identified, their distribution depending mainly on the anthropogenic disturbance of the site. Gigasporaceae Cetraspora sp. and Dentiscutata sp. preferred undisturbed soil whereas Diversisporaceae, Glomeraceae and other Gigasporaceae were associated with recently restored soils. All AMF species were found in all plant species although Corymbiglomus corymbiforme was mainly associated with Echinophora spinosa. Our results might be of help for Mediterranean sand dune restoration.     

The role of genetic and environmental factors in phenotypic variability of arterial pressure and body weight and their interconnection (a family study)

Contribution of genetic and environmental factors into phenotypic variability of blood pressure and body mass as well as into phenotypic correlation between these traits was defined. It was shown that additive genetic determinant is responsible for considerable level of blood pressure and body mass variability. Those common environmental factors are of importance which operate within the limits of one generation. Maternal effect of systolic and diastolic blood pressure variability is negligible. The correlation between systolic and diastolic blood pressure is determined both by environmental and genetic factors, whereas that between blood pressure and body mass is mediated by only environmental factors. The contribution of environmental factors into correlation between blood pressure and body mass lessens with age. The results obtained may be applied in development of population and individual programmes for preventing cardiovascular diseases.     

Geographical patterns of morphological variation and environmental correlates in contact zones: a multi-scale approach using two Mediterranean vipers (Serpentes)

Adaptation to similar selective pressures can explain morphological convergence between closely related species in contact zones. Geostatistics and Geographical Information System were used to identify multi-scale patterns of morphological variability and test the hypothesis of morphological convergence due to local environmental pressures in the contact areas between Vipera aspis and Vipera latastei in the Iberian Peninsula. Nine morphological traits from 630 and 362 vipers at regional and local scale, respectively, were interpolated by Kriging to generate surfaces of morphological variation. Kriging is a geostatistical algorithm that allows investigating the spatial structure of data with statistical models. At both scales, a convergent north–south pattern in morphological variability was observed and the contact areas were identified as integration zones where intermediate vipers are found. Significant correlations were found between surfaces of univariate and multivariate traits, with precipitation and temperature seasonality. Thus, several morphological traits were apparently under local environmental selection. Nevertheless, the influence of biotic pressures and gene flow on morphological convergence of vipers in contact zones deserves further study.     

Population size and structure of three mussel species (Bivalvia: Unionidae) in a northeastern German river with special regard to influences of environmental factors

Unionid mussels are of great ecological importance in running and standing waters, however their populations declined continually during the 20th century. In order to collect more data on the situation of these mussels in running waters, the populations of Anodonta anatina, Anodonta cygnea, and Unio tumidus (Unionidae) were investigated in the northeastern German river Ryck from 1996 to 1998. At three sampling stations along the river, the mussel stocks as well as sediment and water properties were analyzed. In this river abundances of up to 138.7 indiv./m² were detected. The average population densities varied from 10.9 to 34.9 indiv./m². More critically however, the age structures showed significant signs of irregular reproductive success and overaging. Consequently, the mussel stocks are going to decline, and furthermore, U.tumidus should be listed as an endangered species in the Ryck river. Negative influences on bivalves can be expected from sporadically occurring low oxygen and high ion concentrations in the water. But after all, the conditions within the sediments seem to be decisive for the survival of juvenile mussels and thus for the age structure of the populations.     

祁连山东段高原鼢鼠(Eospalax baileyi)土丘空间分布格局及其与环境因子的空间关联性

高原鼢鼠推土造丘行为对高寒草地生态系统的生产和生态功能有重要影响,研究高原鼢鼠土丘空间分布格局及其与环境因子的关系,可以揭示高原鼢鼠栖息地利用和选择规律,为合理控制鼠害及保护草地生物多样性提供科学依据。于2014年8月在祁连山东段选取面积为140m×100m的高原鼢鼠栖息地,消除景观尺度取样带来的气候、地形和土壤的异质性,利用地统计学方法,分析高原鼢鼠土丘的空间分布格局、并揭示其与环境因子中土壤容重、土壤水分、植物地上、地下生物量、根系营养物质含量(可溶性糖、粗蛋白、粗脂肪)以及各功能群丰富度(禾本科、莎草科、杂类草)的空间关系。半方差函数及普通克里格插值表明,高原鼢鼠土丘存在中等程度的空间变异且呈现聚集分布,各环境因子均存在不同程度的空间异质性。交方差函数分析表明,高原鼢鼠分布虽与各环境因子在多种尺度下表现出复杂的空间关联性(正的或负的),但mantel检验发现土壤容重、莎草科丰富度与高原鼢鼠土丘分布呈现显著的负空间关联性,杂类草丰富度和根系粗脂肪含量与高原鼢鼠土丘分布存在显著正空间关联性。综上所述,高原鼢鼠主要栖息利用在土壤疏松、莎草科丰富度较低、杂类草较多和根系粗脂肪含量较高的地方。     

Sediment distribution and accumulation in lagoons of the Southern Mediterranean Region (the MELMARINA Project) with special reference to environmental change and aquatic ecosystems

Surface sediments and sediment cores were collected from coastal lagoons and lakes located in the Southern Mediterranean Region (SMR) as part of the MELMARINA Project which involved integrated eco-hydrological monitoring and modelling. This study uses surface sediments and sediment cores to infer spatial characteristics and temporal changes at the MELMARINA primary sites, Merja Zerga in Morocco, Ghar El Melh in Tunisia and Lake Manzala in Egypt. In addition, surface sediment sampling was undertaken at Egyptian Lake Bardawil and sediment cores were collected from the Lagune de Nador (Morocco). Sediment distribution patterns are investigated using GIS with georeferenced sample locations to facilitate display and resurvey. Major variations in sedimentary organic matter and, particularly, carbonate content, occur within and between sites. Local landscapes combined with hydrological and biogeochemical processes influence the distributions of sediment bulk components (carbonates, organic material and clastic matter) and molluscan shells and shell debris are an important source of sedimentary carbonate at all three primary sites. Sediment cores were dated using natural (²¹⁰Pb) and artificial (¹³⁷Cs) radionuclides, and sediment accumulation rate changes indicate that sources of sediment supply varied markedly through the twentieth century but have generally diminished after the mid-1960s. Sedimentary siliceous microfossils (diatoms) were generally poorly preserved, but mollusc shell remains were well represented. Sediment chronologies and sediment bulk composition allow discussion of some recent changes in bulk, minerogenic and biogenic sediment accumulation patterns in the SMR lagoons. Sediment accumulation rates also varied between sites and multiple cores from Lake Manzala indicated that rates showed considerable spatial variability. Low-level sediment contamination by fossil fuel combustion particulates and trace metals was demonstrated for Ghar El Melh and Lagune de Nador where Pb and Zn accumulation rates were highest in twentieth century sediment. It is emphasized that sediment quality and quantity have strong influences on lagoon ecosystem function and sedimentation is relevant to hydromorphology and to concepts of ecological quality. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Guest editors: J. R. Thompson & R. J. Flower Hydro-ecological Monitoring and Modelling of North African Coastal Lagoons     

Ecogeographic variation of body size in the spectacled salamanders (Salamandrina): influence of genetic structure and local factors

Aim The patterns and causes of ecogeographical body size variation in ectotherms remain controversial. In amphibians, recent genetic studies are leading to the discovery of many cryptic species. We analysed the relationships between body size and climate for a salamander (Salamandrina) that was recently separated into two sibling species, to evaluate how ignoring interspecific and intraspecific genetic structure may affect the conclusions of ecogeographical studies. We also considered the potential effects of factors acting at a local scale. Location Thirty‐four populations covering the whole range of Salamandrina, which is endemic to peninsular Italy. Methods We pooled original data and data from the literature to obtain information on the snout–vent length (SVL) of 3850 Salamandrina females; we obtained high‐resolution climatic data from the sampled localities. We used an information‐theoretic approach to evaluate the roles of climate, genetic features (mitochondrial haplogroup identity) and characteristics of aquatic oviposition sites. We repeated our analyses three times: in the first analysis we ignored genetic data on intraspecific and interspecific variation; in the second one we considered the recently discovered differences between the two sibling species; in the third one we included information on intraspecific genetic structure within Salamandrina perspicillata (for Salamandrina terdigitata the sample size was too small to perform intraspecific analyses). Results If genetic information was ignored, our analysis suggested the existence of a relationship between SVL and climatic variables, with populations of large body size in areas with high precipitation and high thermal range. If species identity was included in the analysis, the role of climatic features was much weaker. When intraspecific genetic differences were also considered, no climatic feature had an effect. In all analyses, local factors were important and explained a large proportion of the variation; populations spawning in still water had a larger body size. Main conclusions An imperfect knowledge of species boundaries, or overlooking the intraspecific genetic variation can strongly affect the results of analyses of body size variation. Furthermore, local factors can be more important than the large‐scale parameters traditionally considered, particularly in species with a small range.     

Importance of short-term dynamics in carbon isotope ratios of ecosystem respiration (delta13C(R)) in a Mediterranean oak woodland and linkage to environmental factors

Temporal dynamics in carbon isotope ratios of ecosystem respiration (delta13C(R)) were evaluated on hourly, daily and annual timescales in a Mediterranean woodland. Emphasis was given to the periods of transition from wet to dry season and vice versa, when the system turns from a net carbon sink to a source. The constancy of nocturnal delta13C(R) was tested. The relationship between delta13C(R) (determined through Keeling plots) and environmental factors was evaluated through time-lag analysis. Delta13C(R) exhibited high annual variation (> 7). During the transition periods, delta13C(R) correlated significantly with factors influencing photosynthetic discrimination, soil respiration, and whole-canopy conductance. Time-lags differed between below- and above-ground variables, and between seasons. A shift in regression parameters with environmental factors indicated seasonal differences in ecosystem responsiveness (e.g. temperature acclimation). Delta13C(R) exhibited substantial nocturnal enrichment (> 4) from dusk to dawn. These data indicate pronounced short-term dynamics in delta13C(R) at hourly to daily timescales and a modulated response to environmental drivers. Substantial short-term changes in nocturnal delta13C(R) may have important implications for the sampling protocols of nocturnal Keeling plots.     

The influence of temperature, relative humidity and host factors on the attachment and survival of Boophilus microplus (Canestrini) larvae to skin slices

Attachment of B. microplus larvae was examined using slices (0.5 mm) of bovine skin stretched over a suitable medium. Optimal temperature for attachment lay between 31 and 38°C. At 38°C, 70 to 80% had attached by 4 h but this was followed 6–7 h after the release of the larvae by a significant decrease in the percentage attached. By 8 h, the percentage attached had risen again and remained at 70–80% for the subsequent 16 h. Attachment was not influenced by ambient relative humidity within the range 20–75 % on the time scale studied (16 h). Most larvae denied access to the skin surface at 20 or 45 % R.H. (38°C) died within 24 h. Those allowed to feed survived at 45% but most died at 20% R.H. There was no difference in attachment when bovine or rabbit serum or phosphate buffered saline was used, nor when skin was taken from Zebu × European cattle or Herefords. Attachment was no different when skin was taken from neck, rib or rump, or from cattle with different levels of resistance to B. microplus but there was reduced attachment with mouse skin. Attachment was reduced on skin which had been stored at ? 15°C for 1 day but storage at 10°C for 16 h had no effect. These results are related to problems of tick water balance, host specificity and stimuli for tick attachment.     

The genetic structure of the loggerhead sea turtle (Caretta caretta) in the Mediterranean as revealed by nuclear and mitochondrial DNA and its conservation implications

The population genetic structure of the loggerhead sea turtle (Caretta caretta) nesting in the eastern Mediterranean was assessed by sequencing a fragment of the control region of the mitochondrial DNA (n = 190) and seven microsatellites (n = 112). The two types of markers revealed genetic structuring (mtDNA: γ_st = 0.212, P < 0.001; nDNA F _st = 0.006, P < 0.001), thus indicating that both females and males are philopatric and that gene flow between populations is restricted. Mitochondrial DNA data indicate that the female populations nesting on the islands of Crete and Cyprus have suffered a recent bottleneck or colonization event. However, no bottleneck or founder effect was revealed by nuclear markers, thus indicating male-mediated gene flow from other populations that would increase nuclear genetic variability. Crete, and to a lower extent Cyprus, are thought to play a central role in such male-mediated gene flow that may reduce the negative effect of genetic drift or inbreeding on the small populations of Lebanon and Israel. This population structure indicates that assessing population relevance only on the basis of genetic variability and size would be misleading, as some populations not fulfilling those requirements may play a relevant role in genetic exchange and hence contribute to the overall genetic variability.