Incidence of human papilloma virus in esophageal squamous cell carcinoma in patients from the Lublin region

AIM:To assess the prevalence of human papilloma virus(HPV) in esophageal squamous cell carcinoma(ESCC) in the south-eastern region of Poland.METHODS:The study population consisted of 56 ESCC patients and 35 controls.The controls were patients referred to our department due to other nonesophageal and non-oncological disorders with no gross or microscopic esophageal pathology as confirmed by endoscopy and histopathology.In the ESCC patients,samples were taken from normal mucosa(56 mucosa samples) and from the tumor(56 tumor samples).Tissue samples from the controls were taken from normal mucosa of the middle esophagus(35 control samples).Quantitative determination of DNA was carried out using a spectrophotometric method.Genomic DNA was isolated using the QIAamp DNA Midi Kit.HPV infection was identified following PCR amplification of the HPV gene sequence,using primers MY09 and MY11 complementary to the genome sequence of at least 33 types of HPV.The sequencing results were computationally analyzed using the basic local alignment search tool database.RESULTS:In tumor samples,HPV DNA was identified in 28 of 56 patients(50%).High risk HPV phenotypes(16 or/and 18) were found in 5 of 56 patients(8.9%),low risk in 19 of 56 patients(33.9%) and other types of HPV(37,81,97,CP6108) in 4 of 56 patients(7.1%).In mucosa samples,HPV DNA was isolated in 21 of 56 patients(37.5%).High risk HPV DNA was confirmed in 3 of 56 patients(5.3%),low risk HPV DNA in 12 of 56 patients(21.4%),and other types of HPV in 6 of 56 patients(10.7%).In control samples,HPV DNA was identified in 4 of 35 patients(11.4%) with no high risk HPV.The occurrence of HPV in ESCC patients was significantly higher than in the controls [28 of 56(50%) vs 4 of 35(11.4%),P < 0.001].In esophageal cancer patients,both in tumor and mucosa samples,the predominant HPV phenotypes were low risk HPV,isolated 4 times more frequently than high risk phenotypes [19 of 56(33.9%) vs 5 of 56(8.9%),P < 0.001].A higher prevalence of HPV was identified in female patients(71.4% vs 46.9%).Accordingly,the high risk phenotypes were isolated more frequently in female patients and this difference reached statistical significance [3 of 7(42.9%) vs 2 of 49(4.1%),P < 0.05].Of the pathological characteristics,only an infiltrative pattern of macroscopic tumor type significantly correlated with the presence of HPV DNA in ESCC samples [20 of 27(74.1%) vs 8 of 29(27.6%) for ulcerative or protruding macroscopic type,P < 0.05].The occurrence of total HPV DNA and both HPV high or low risk phenotypes did not significantly differ with regard to particular grades of cellular differentiation,phases in depth of tumor infiltration,grades of nodal involvement and stages of tumor progression.CONCLUSION:Low risk HPV phenotypes could be one of the co-activators or/and co-carcinogens in complex,progressive,multifactorial and multistep esophageal carcinogenesis.

Genetic Diversity of Italian Ryegrass Diploid Cultivars, Revealed by Electrophoretically Detected Genotypes in Phosphoglucose Isomerase （PGI）

Nine diploid cultivars of Italian ryegrass （Lolium multiflorum Lain.） from France （Fortyl, Vertyl and Jericho）, Germany （Ligrande）, United Kingdom （Aber Epic and Aber Mario）, Denmark （Cordelia）, Netherlands （Alamo） and Poland （Tur） were tested with horizontal gel electrophoresis according to one locus （with four alleles） of the PGI enzyme system. One of them, named P4 is typical for the species, therefore can serve as a good marker for hybrids identification. Each cultivar was characterized by frequencies of different phenotypes. They were highly polymorphic （Pg = 0.58 - 0.78） and showed differences in heterozygosity level. The variability within populations （GST = 0.055） was higher than among populations （DST = 0.032）.

Genetic Structure of Tetraploid Italian and Westerwolds Ryegrasses （Lolium spp,） as Revealed by Enzyme and ISSR Polymorphism

The genetic variation of Italian ryegrass （Lolium multiflorum Lain.） and westerwolds ryegrass （L. westerwoldicum Breakw.） was investigated in eight tetraploid cultivars originated from Poland. The 337 individual seedlings, cultivated in uniform greenhouse conditions, were tested with horizontal gel electrophoresis according to five enzyme systems： diaforase （DIA）, superoxide dismutase （SOD）, glutamate oxalacetate transaminase （GOT）, peroxidise （PX） and phosphogluco isomerise （PGI）, which were used to array allelic diversity at five polymorphic loci. The estimation of genetic diversity in 64 plants of the same cultivars was studied using inter simple sequence repeats （ISSRs） as molecular marker. The genetic variation described by ISSR suggests that the polymorphism detected appears to be poorly informative at the taxonomic level. For statistical analysis, unweighted pair group method with arithmetic mean （UPGMA） based dendrograms and minimum spanning trees were constructed using Nei＇s distance and Jaccard＇ s similarity coefficient.

Application of Molecular Genetic Methods in Anthropological and Paleodemographic Studies of Fragmentary and Damaged Skeletal Material from Rescue Excavations

The purpose of this study was to demonstrate the application of molecular genetic methods in anthropology and paleodemography in case where the examined bone material is damaged and fragmented, and where the skeletal remains of children are investigated. The application of traditional anthropological methods is limited, especially if sex determination of children and fragmentary skeletal remains is detected. Sex typing genetic markers （SRY, amelogenin） were used for sex determination of children and undetermined fragmentary skeletal remains from the burial site Pohansko, south outer precincts （Czech Republic）. This is an approximately 1,200 year-old burial site （the Great Moravian period）, which was excavated during rescue excavations. After the genetic analysis, sex was determined in more than half of the investigated samples. The results of the genetic analysis were used for completion of demographic data of this archaeological site. The results of sex determination of several samples were independently verified by the Institute of Criminology in Prague （Czech Republic）, using the PowerPlex ESX 17 System （Promega）. This study showed the suitability of modem molecular genetic methods to skeletal anthropology and paleodemographic analyses.

The Impact of Urban Noise on the Behavior of Two Mouse Species Belonging to the Genus Apodemus

Traffic noise was tested as a stress factor in an open-field study of urban and non-urban populations of two species belonging to the genus Apodemus: the striped field mouse and the yellow-necked mouse. The striped field mouse has inhabited green areas in cities for years, whereas the yellow-necked mouse was first observed in such areas relatively recently. Behavioral reactions to noise in the urban population of striped field mice were the same as the reactions observed in the non-urban population of the species. As for the yellow-necked mouse, behavior in response to noise in the non-urban population differed from that observed in the urban population. We see such differences as being related to the fact that striped field mice, which have inhabited urban areas for many generations, form a stable population in urban areas, and specific individuals are adapted to urban conditions. Yellow-necked mice, in turn, form no stable urban population and the individuals found in cities are most probably migrants, not adapted to urban conditions, which is the reason why their reaction to noise differs from the typical response.

大鼠小胶质细胞在浸润前或炎症极化后的开放染色质区域数据

小胶质细胞是脑部驻留的骨髓系细胞,在生理和病理情况下的大脑中均起重要作用。本研究报道了关于大鼠小胶质细胞DNA酶超敏(DHS)的开放染色质区域的数据集。该数据集全面、可复制、错误发现率可控。我们用未处理的和神经胶质瘤条件培养基(GCM)或脂多糖(LPS)刺激6 h的原代培养小胶质细胞,比较了小胶质细胞开放染色质图谱。结果显示胶质瘤分泌因子诱导小神经胶质细胞的侵袭性和免疫抑制性激活,而激活的小胶质细胞促进肿瘤生长。大鼠小胶质细胞的开放染色质图谱由126640个可重复的DHS区域组成,其中分别有2303和12357个区域在用GCM或LPS刺激后显示出开放性的显著变化。活性基因表现出组成型开放的启动子,但基因附近的DHS区域的聚集开放性与该基因表达之间没有直接依赖关系。对应同一基因的各个区域经常呈现不同的开放性变化模式。GCM调节的DHS区域更多出现在远离基因体的区域,而LPS调节的区域更多出现在内含子中。GCM和LPS对不同区域的开放性调控的差异与免疫检查点基因相对应。这两种处理在对Toll样受体信号传导和轴突导向途径中对应基因区域的聚集开放性有不同的影响,这表明小胶质细胞迁移所使用的分子机制与轴突生长相似,并且这些通路的调节有助于胶质瘤诱导的小胶质细胞侵袭性极化。我们的开放染色质区域数据集为大鼠小胶质细胞中基因调控的研究铺平了道路。

Multitasking guardian of mitochondrial quality:Parkin function and Parkinson’s disease

The familial form of Parkinson's disease(PD)is linked to mutations in specific genes.The mutations in parkin are one of the most common causes of early-onset PD.Mitochondrial dysfunction is an emerging active player in the pathology of neurodegenerative diseases,because mitochondria are highly dynamic structures integrated with many cellular functions.Herein,we overview and discuss the role of the parkin protein product,Parkin E3 ubiquitin ligase,in the cellular processes related to mitochondrial function,and how parkin mutations can result in pathology in vitro and in vivo.

SOX9-transactived long non-coding RNA NEAT1 promotes the self-renewal of liver cancer stem cells through PKA/Hippo signaling

Dear Editor,Hepatocellular carcinoma(HCC),the most common pathological type of primary liver cancer,ranks as the third deadliest cancer.Despite the progress of surgical resection in recent years,the 5-year survival of HCC patients is still unsatisfactory due to the frequent relapse and chemoresistance.Accumulating evidence has demonstrated that liver cancer stem cells(CSCs)are critical for HCC chemoresistance and recurrence.Nevertheless,the molecular mechanisms of liver CSC regulation remain unclear,which hampers the development of the therapeutic strategy that targets liver CSCs.

Brain GABA and glutamate levels in workers of two ant species （Hymenoptera： Formicidae）： Interspecific differences and effects of queen presence/absence

Presence of amino acid neurotransmitters gamma-aminobutyric acid （GABA） and glutamate （Glu） in ant brains was reported in very few studies. To learn more about factors influencing GABA and Glu levels in ant brains, we applied high-performance liquid chromatography to measure levels of these compounds in single brains of workers of 2 ant species, Myrmica ruginodis （subfamily Myrmicinae） and Formica polyctena （subfamily Formicinae） taken from queenright/queenless colony fragments and tested in dyadic aggression tests consisting of an encounter with a nestrnate, an alien conspecific or a small cricket. Brain glutamate levels were higher than those of GABA in both tested species. Brain GABA levels （in μmol/brain） and GABA ： Glu ratio were higher in M. ruginodis （a submissive species） than in E polyctena （a dominant, aggressive species） in spite of smaller brain weight of M. ruginodis. Brain glutamate levels （in μmol/brain） did not differ between the tested species, which implies that glutamate concentration （in μmol/mg of brain tissue） was higher in M. ruginodis. Queen absence was associated with increased worker brain GABA levels in E polyctena, but not in M. ruginodis. No significant effects of opponent type were discovered. As GABA agonists enhance friendly social behavior in rodents, we hypothesize that elevated brain GABA levels of orphaned workers of F.polyctena facilitate the adoption of a new queen. This is the first report providing information on GABA and glutamate levels in single ant brains and documenting the effects of queen presence/absence on brain levels of amino acid neurotransmitters in workers of social Hymenoptera.

Sustained activation of P2X7 induces MMP-2- evoked cleavage and functional purinoceptor inhibition

P2X7 purinoceptor promotes survival or cytotoxicity depending on extraceUular adenosine triphosphate （ATP） stimulus Intensity controlling its ion channel or P2X7-dependent large pore （LP） functions. Mechanisms governing this operational divergence and functional idiosyncrasy are ill-understood. We have discovered a feedback loop where sustained activation of P2X7 triggers release of active matrix metalloproteinase 2 （MMP-2）, which halts ion channel and LP responses via the MMP-2-dependent receptor cleavage. This mechanism operates in cells as diverse as macrophages, dystrophic myoblasts, P2X7-transfected HEK293, and human tumour cells. Given that serum-born MMP-2 activity also blocked receptor functions, P2X7 responses in vivo may decrease in organs with permeable capillaries. Therefore, this mechanism represents an Important fine-tuning of P2X7 functions, reliant on both cell-autonomous and extraneous factors. Indeed, it allowed evasion from the ATP-induced cvtotoxicity in macrophages and human cancer ceils with high P2X7 expression levels. Finally, we demonstrate that P2X7 ablation eliminated getatinase activity in inflamed dystrophic muscles in vivo. Thus, P2X7 antagonists could be used as an alternative to highly toxic MMP inhibitors in treatments of inflammatory diseases and cancers.

Metabolic regulation of microglial phagocytosis:Implications for Alzheimer’s disease therapeutics

Microglia,the resident immune cells of the brain,are increasingly implicated in the regulation of brain health and disease.Microglia perform multiple functions in the central nervous system,including surveillance,phagocytosis and release of a variety of soluble factors.Importantly,a majority of their functions are closely related to changes in their metabolism.This natural inter-dependency between core microglial properties and metabolism offers a unique opportunity to modulate microglial activities via nutritional or metabolic interventions.In this review,we examine the existing scientific literature to synthesize the hypothesis that microglial phagocytosis of amyloid beta(Aβ)aggregates in Alzheimer’s disease(AD)can be selectively enhanced via metabolic interventions.We first review the basics of microglial metabolism and the effects of common metabolites,such as glucose,lipids,ketone bodies,glutamine,pyruvate and lactate,on microglial inflammatory and phagocytic properties.Next,we examine the evidence for dysregulation of microglial metabolism in AD.This is followed by a review of in vivo studies on metabolic manipulation of microglial functions to ascertain their therapeutic potential in AD.Finally,we discuss the effects of metabolic factors on microglial phagocytosis of healthy synapses,a pathological process that also contributes to the progression of AD.We conclude by enlisting the current challenges that need to be addressed before strategies to harness microglial phagocytosis to clear pathological protein deposits in AD and other neurodegenerative disorders can be widely adopted.

Stem Cell Regulation by Arabidopsis WOX Genes

Gene amplification followed by functional diversification is a major force in evolution. A typical example of this is seen in the WUSCHEL-RELATED HOMEOBOX （WOX） gene family, named after the Arabidopsis stem cell regulator WUSCHEL. Here we analyze functional divergence in the WOX gene family. Members of the WUS clade, except the cambium stem cell regulator WOX4, can substitute for WUS function in shoot and floral stem cell maintenance to different degrees. Stem cell function of WUS requires a canonical WUS-box, essential for interaction with TPL/TPR co-repressors, whereas the repressive EAR domain is dispensable and the acidic domain seems only to be required for female fertility. In contrast to the WUS clade, members of the ancient WOX13 and the WOX9 clades cannot support stem cell maintenance. Although the homeodomains are interchangeable between WUS and WOX9 clade members, a WUS- compatible homeodomain together with canonical WUS-box is not sufficient for stem cell maintenance. Our results suggest that WOX function in shoot and floral meristems of Arabidopsis is restricted to the modern WUS clade, suggesting that stem cell control is a derived function. Yet undiscovered functional domains in addition to the homeodomain and the WUS-box are necessary for this function.

FURTHER STUDIES ON THE MESODERM-INDUCING SUBSTANCE FROM PIG LIVER

In our previous work we have reported the procurement of a mesoderminducing protein from pig liver, which, as evidenced by electrophoretic findings, was still contaminated with impurities. Recently, the material was further purified by CM cellulose column chromatography with continuous gra-