Rac1 Regulates Endometrial Secretory Function to Control Placental Development

During placenta development, a succession of complex molecular and cellular interactions between the maternal endometrium and the developing embryo ensures reproductive success. The precise mechanisms regulating this maternal-fetal crosstalk remain unknown. Our study revealed that the expression of Rac1, a member of the Rho family of GTPases, is markedly elevated in mouse decidua on days 7 and 8 of gestation. To investigate its function in the uterus, we created mice bearing a conditional deletion of the Rac1 gene in uterine stromal cells. Ablation of Rac1 did not affect the formation of the decidua but led to fetal loss in mid gestation accompanied by extensive hemorrhage. To gain insights into the molecular pathways affected by the loss of Rac1, we performed gene expression profiling which revealed that Rac1 signaling regulates the expression of Rab27b, another GTPase that plays a key role in targeting vesicular trafficking. Consequently, the Rac1-null decidual cells failed to secrete vascular endothelial growth factor A, which is a critical regulator of decidual angiogenesis, and insulin-like growth factor binding protein 4, which regulates the bioavailability of insulin-like growth factors that promote proliferation and differentiation of trophoblast cell lineages in the ectoplacental cone. The lack of secretion of these key factors by Rac1-null decidua gave rise to impaired angiogenesis and dysregulated proliferation of trophoblast cells, which in turn results in overexpansion of the trophoblast giant cell lineage and disorganized placenta development. Further experiments revealed that RAC1, the human ortholog of Rac1, regulates the secretory activity of human endometrial stromal cells during decidualization, supporting the concept that this signaling G protein plays a central and conserved role in controlling endometrial secretory function. This study provides unique insights into the molecular mechanisms regulating endometrial secretions that mediate stromal-endothelial and stromal-trophoblast crosstalk critical for placenta development and establishment of pregnancy.     

Size factor as one of the problems in the sex estimation of skulls: upper palaeolithic sample from Predmostí (Czech Republic) as an example

In this study we estimate sex and population affinity of Gravettian Predmostí (P) skulls using linear and geometric discriminant analysis (DA), and compare them with results of 2D geometric morphometrics (GM). We used the measurements of P3 and P9 males, P4 and P10 females, as originally estimated by Matiegka (1934), as well as two databases--the recent skull database of Howells and the fossil data of Henke. DA classifies the P skulls as robust and belonging to the "male" region, loosing the sensitivity of inter-population differences influenced by size factor. That is why this approach could not be applied. The geographic inter-population differences according to DA do not define P skull shapes as extreme. The influence of geographic variability could be stronger than the inter-sexual differences. Despite the chronological differences between databases and Gravettian skulls, these differences are a component of regional inter-population variability. According to our results, GM is more successful methodological approach than DA. Our previous sex estimation of P skulls with help of GM is completely in accordance with the classical morphoscopic estimation. However, an appropriate reference database is necessary in both the GM and DA approaches. For the sexing of skull with unknown population affinity, and with absence of appropriate reference database, we suggest to use the application of more subjective visual scoring methods.     

Arsenic in Agricultural Soils of a Historically Mined and Industrial Region of Southern Serbia and Northern Kosovo: Bioavailability and Uptake by Plants Species Zea mays L. and Solanum tuberosum L.

This article reports the results of a study focused on the presence and bioavailability of arsenic in agricultural soil in the mining and industrial regions of northern Kosovo and southern Serbia, as well as uptake and bioaccumulation of arsenic in two commonly cultivated plant species (Zea mays L. and Solanum tuberosum L.). This area was one of the most important mining districts in Europe. The collected soil samples were subjected to a modified BCR three-step sequential extraction procedure in order to investigate the chemical partitioning of arsenic in the soils. The general distribution of arsenic in various fractions was: exchangeable < reducible < oxidizable fractions. Highest concentrations of total arsenic in soil were found close to industrial facilities and tailing ponds. In addition, fluvisols were significantly more enriched with arsenic than soils at a distance from the river flows. The edible parts of the plant specimen showed different As contents, suggesting that these plant species have different propensities for the uptake and bioaccumulation of arsenic from soil.     

Integrative taxonomy and evolutionary history of a newly revealed spider Dysdera ninnii complex (Araneae: Dysderidae)

The spider genus Dysdera is a species‐rich clade of specialized woodlice predators, composed typically of complexes of sibling species. Here, we analyse the Dysdera ninnii complex, distinguishing three species that exhibit slight but constant differences in the morphology of their copulatory organs, and in their genetic background. We designate a neotype for D. ninnii and redescribe it. We consider Dysdera pavesii Thorell, 1873 to be a junior synonym of Dysdera ninnii Canestrini, 1868. In addition, we describe two new species ( D ysdera moravica sp. nov. and D ysdera microdonta sp. nov. ). All three species occur in the region of north‐eastern Italy, Slovenia, and north‐western Croatia. D ysdera moravica sp. nov. expanded to central Europe. The species occur allopatrically or parapatrically. All three species possess the same diploid number and X0 sex chromosome determination. In some individuals we found chromosome fusions, and such polymorphism is common in spiders with holokinetic chromosomes. The analysis of mitochondrial (cytochrome c oxidase subunit I, COI) and nuclear ribosomal (internal transcribed spacer 2, ITS2) DNA markers revealed two clades, one formed by D. ninnii and D . microdonta sp. nov. , and a second by D . moravica sp. nov. Species of the first clade are not well defined by DNA markers. We noticed only weak separation of maternally inherited COI, and even overlap of autosomally inherited ITS2 sequences. We suggest that either short speciation time, unfinished lineage sorting, or rare hybridization events caused this pattern. In one sample of D . microdonta sp. nov. we detected the coxA gene of a Rickettsia species, which is the first record of this parasitic bacteria from the spider family Dysderidae. D ysdera microdonta sp. nov. occurs at higher altitudes than D. ninnii, and their distribution ranges form a long contact zone. Remarkably, we did not record any overlap of the two distribution ranges, suggesting that the lack of a precopulatory interspecific barrier causes a loss of reproduction potential. We hypothesize that because of the unsolved interspecific barrier together with only tiny differences in morphology and COI sequences, and no differences in karyotypes and ITS2 sequences, the D. ninnii species complex is evolutionarily young. © 2014 The Linnean Society of London     

不同方法处理的九香虫对人胃癌SGC-7901细胞体外作用比较及其抑癌组分分布

本实验探讨冷冻和传统中药炮制方法处理的九香虫对胃癌细胞增殖的影响及九香虫抑癌活性组分的体内分布。体外培养人胃癌SGC-7901细胞,采用四甲基偶氮唑盐比色法(MTT法)观察不同方法处理的九香虫各组分水溶液对SGC-7901细胞的体外抑制作用。结果发现,炮制九香虫蛋白浓度为50和100mg·L~(-1)时作用48h对SGC-7901细胞生长具有抑制作用,抑制率分别为13.45%和14.68%,而浓度达到200和400mg·L~(-1)时对SGC-7901细胞生长具有促进作用,抑制率为-7.94%和-82.50%;冷冻处理下九香虫不同浓度对SGC-7901细胞生长具有显著的抑制作用,该处理组蛋白浓度为50、100、200和400mg·L-1时抑制率分别为0.49%,3.82%,4.42%,39.33%。选取九香虫整虫及分解后的各部位处理组最大作用浓度比较,增殖抑制率为血淋巴腹部整虫头部。因此,冷冻处理的九香虫对胃癌细胞抑制率更高,且在该条件下九香虫的抑癌活性组分主要分布于血淋巴和腹部。     

Dosimetric accuracy of three dose calculation algorithms for radiation therapy of in situ non-small cell lung carcinoma

BackgroundStudy determines differences in calculated dose distributions for non-small cell lung carcinoma (NSC LC) patients. NSC LC cases were investigated, being the most common lung cancer treated by radiotherapy in our clinical practice.Materials and methodsA retrospective study of 15 NSCLC patient dose distributions originally calculated using standard superposition (SS) and recalculated using collapsed cone (CC ) and Monte Carlo (MC) based algorithm expressed as dose to medium in medium (MCD_m) and dose to water in medium (MCD_w,) was performed so that prescribed dose covers at least 99% of the gross target volume (GTV). Statistical analysis was performed for differences of conformity index (CI), heterogeneity index (HI), gradient index (GI), dose delivered to 2% of the volume (D_2%), mean dose (D_mean) and percentage of volumes covered by prescribed dose (V_70Gy). For organs at risk (OARs), D_mean and percentage of volume receiving 20 Gy and 5Gy (V_20Gy, V_5Gy) were analysed.ResultsStatistically significant difference for GTVs was observed between MCD_w and SS algorithm in mean dose only. For planning target volumes (PTVs), statistically significant differences were observed in prescribed dose coverage for CC, MCD_m and MCD_w. The differences in mean CI value for the CC algorithm and mean HI value for MCD_m and MCD_w were statistically significant. There is a statistically significant difference in the number of MUs for MCD_m and MCD_w compared to SS.ConclusionAll investigated algorithms succeed in managing the restrictive conditions of the clinical goals. This study shows the drawbacks of the CC algorithm compared to other algorithms used.     

Influence of Phospholipids on Growth, Sporulation and Virulence of the Endoparasitic Fungi Drechmeria coniospora, Verticillium balanoides and Harposporium anguillulae in Liquid Culture

Verticillium balanoides mycelial growth was stimulated on solid corn meal agar (1.7 %) and in liquid corn meal broth (0.2 %) upon the addition of phospholipids at various concentrations. Sporulation differed with phospholipid products and was highest in pure corn meal. Drechmeria coniospora mycelial growth increased upon addition of phospholipids at all concentrations in solid or liquid culture. Sporulation increased at high concentration (1000 ppm) and decreased at low concentration (100 ppm) of phospholipids in the medium. For both fungi, infectivity of conidia produced in liquid culture decreased when compared to conidia from parasitized nematodes. Addition of phospholipids partly restored this effect. Harposporium anguillulae mycelial growth and sporulation was not affected by addition of phospholipids to solid or liquid corn meal medium.     

(2-Benzyl-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indol-8-yl)-acetic acid: an aldose reductase inhibitor and antioxidant of zwitterionic nature

Novel carboxymethylated pyridoindoles, characterized by antioxidant activity combined with the ability to inhibit aldose reductase, represent an example of a multitarget approach to the treatment of diabetic complications - severe diabetes-related health disorders of multifunctional nature. One of the novel carboxymethylated pyridoindoles, (2-benzyl-2,3,4,5-tetrahydro-1H-pyrido[4,3-b]indol-8-yl)-acetic acid (compound 1), was found to inhibit aldose reductase with the IC(50) value 18.2 ± 1.2 μM. Owing to aldose reductase pharmacophore requirements for an acidic proton, most aldose reductase inhibitors contain an acetic acid moiety, ionized at physiological pH, resulting in poor bioavailability of the drugs. The presence of a basicity center at the tertiary nitrogen of the carboxymethylated pyridoindoles, in addition to the acidic carboxylic function, predisposes these compounds to form double-charged zwitterionic species. The zwitterionic nature of compound 1 may remarkably affect its pH-lipophilicity profile allowing for increased membrane penetration in the pH region around its isoelectric point, which lies close to the physiological pH 7.4. In the first part of this study, the presence of zwitterionic species was experimentally proved by the concentration-dependent effect of sodium 1-hexanesulphonate on the distribution profile of compound 1. Then a series of experiments was performed in the cellular system of isolated erythrocytes in vitro. Isolated rat erythrocytes exposed to peroxyl radicals, generated in the solution by decomposition of the hydrophilic azoinitiator AAPH or intracellularly by decay of lipophilic t-BuOOH, underwent progressive hemolysis. The onset of the hemolysis was shifted from the starting zero point by the time interval assigned as a lag period. In the presence of compound 1 the lag period was significantly prolonged. Finally, under conditions of a short-term experiment in STZ-diabetic rats in vivo, increase in sorbitol levels in erythrocytes was recorded. Compound 1 administered in the dose 50mg/kg/day (i.g.) significantly decreased the sorbitol level in the erythrocytes. To conclude, the physico-chemical proof of the zwitterionic nature of compound 1 was established and the results obtained in isolated red blood cells indicated good cellular availability of the compound. In addition, in diabetic rats, sorbitol accumulation in red blood cells was significantly inhibited by compound 1 administered intra-gastrically, suggesting its ready uptake into the central compartment. The zwitterionic principle thus may have significant consequences for increased bioavailability of drugs bearing an acidic function.     

Endoplasmic Reticulum Stress Induced Synthesis of a Novel Viral Factor Mediates Efficient Replication of Genotype-1 Hepatitis E Virus

Hepatitis E virus (HEV) causes acute hepatitis in many parts of the world including Asia, Africa and Latin America. Though self-limiting in normal individuals, it results in ~30% mortality in infected pregnant women. It has also been reported to cause acute and chronic hepatitis in organ transplant patients. Of the seven viral genotypes, genotype-1 virus infects humans and is a major public health concern in South Asian countries. Sporadic cases of genotype-3 and 4 infection in human and animals such as pigs, deer, mongeese have been reported primarily from industrialized countries. Genotype-5, 6 and 7 viruses are known to infect animals such as wild boar and camel, respectively. Genotype-3 and 4 viruses have been successfully propagated in the laboratory in mammalian cell culture. However, genotype-1 virus replicates poorly in mammalian cell culture and no other efficient model exists to study its life cycle. Here, we report that endoplasmic reticulum (ER) stress promotes genotype-1 HEV replication by inducing cap-independent, internal initiation mediated translation of a novel viral protein (named ORF4). Importantly, ORF4 expression and stimulatory effect of ER stress inducers on viral replication is specific to genotype-1. ORF4 protein sequence is mostly conserved among genotype-1 HEV isolates and ORF4 specific antibodies were detected in genotype-1 HEV patient serum. ORF4 interacted with multiple viral and host proteins and assembled a protein complex consisting of viral helicase, RNA dependent RNA polymerase (RdRp), X, host eEF1α1 (eukaryotic elongation factor 1 isoform-1) and tubulinβ. In association with eEF1α1, ORF4 stimulated viral RdRp activity. Furthermore, human hepatoma cells that stably express ORF4 or engineered proteasome resistant ORF4 mutant genome permitted enhanced viral replication. These findings reveal a positive role of ER stress in promoting genotype-1 HEV replication and pave the way towards development of an efficient model of the virus.     

Attachment style in parents of children with chronic gastrointestinal disease

Attachment is a point of interest in psychosomatic research since it influences a wide array of biopsychosocial phenomena. Data from literature highlights the role of this concept in the context of Inflammatory Bowel disease (IBD), still, there is a lack of data regarding attachment among parents of children with chronic gastrointestinal diseases. The main hypothesis for the current study is that parents of children with IBD will have a more insecure attachment than parents of children with celiac disease (CD) and parents of healthy children. The second hypothesis is that insecure attachment among parents of sick children will be associated with lower parental quality of life (QoL). 46 parents of children with IBD, 42 parents of children with CD and 43 parents of healthy children completed the validated modification of the Brennan's Experiences in Close Relationship Inventory. Results were categorized as secure and insecure attachment. In order to assess parental QoL, the WHOQOL-BREF questionnaire was used. The Total QoL was calculated as a sum of all domain items. Secure attachment was found in 45.7% parents of children with IBD, in 35.7% parents of children with CD and in 32.6% parents of healthy children. Surprisingly, the lowest rate of secure attachment was found in parents of healthy children. However, significant differences among groups do not exist. For all groups of parents the attachment style is associated with Total QoL, although only among parents of children with IBD, the secure attachment independently and significantly predicts higher parental Total QoL. According to results, we might say that parental attachment style does not have a role that exclusively belongs in the context of paediatric chronic gastrointestinal diseases. However, parents of children with IBD who have insecure attachment represent target group for psychosocial support in order to improve their QoL.