Expression of R-spondins/Lgrs in development of movable craniofacial organs

Wnt signaling plays a critical role in the development of many organs, including the major movable craniofacial organs tongue, lip, and eyelid. Four members of the R-spondin family (Rspo1–4) bind to Lgr4/5/6 to regulate the activation of Wnt signaling. However, it is not fully understood how Rspos/Lgrs regulate Wnt signaling during the development of movable craniofacial organs. To address this question, we examined the expression of Rspos, Lgrs, and Axin2 (major mediator of canonical Wnt signaling) during tongue, lip, and eyelid development. The expression of Axin2, Rspos and Lgrs was observed in many similar regions, suggesting that Rspos likely activate canonical Wnt signaling through the Lgr-dependent pathway in these regions. Lgr expression was not detected in regions where Axin2 and Rspos were expressed, suggesting that Rspos might activate canonical Wnt signaling through the Lgr-independent pathway in these regions. In addition, the expression of Rspos and Lgrs were observed in some other regions where Axin2 was not expressed, suggesting the possibility that Rspos and/or Lgrs are involved in non-canonical Wnt signaling or the Wnt-independent pathway. Thus, we identified a dynamic spatiotemporal expression pattern of Rspos and Lgrs during the development of the eyelid, tongue, and lip.     

Comparison of sand fly trapping approaches for vector surveillance of Leishmania and Bartonella species in ecologically distinct,endemic regions of Peru

BackgroundIn Peru, the information regarding sand fly vectors of leishmaniasis and bartonellosis in the Amazon region is limited. In this study, we carried out sand fly collections in Peruvian lowland and highland jungle areas using different trap type configurations and screened them for Leishmania and Bartonella DNA.Methodology/Principal findingsPhlebotomine sand flies were collected in Peruvian Amazon jungle and inter Andean regions using CDC light trap, UV and color LED traps, Mosquito Magnet trap, BG Sentinel trap, and a Shannon trap placed outside the houses. Leishmania spp. screening was performed by kDNA PCR and confirmed by a nested cytochrome B gene (cytB) PCR. Bartonella spp. screening was performed by ITS PCR and confirmed by citrate synthase gene (gltA). The PCR amplicons were sequenced to identify Leishmania and Bartonella species.UV and Blue LED traps collected the highest average number of sand flies per hour in low jungle; UV, Mosquito Magnet and Shannon traps in high jungle; and Mosquito Magnet in inter Andean region. Leishmania guyanensis in Lutzomyia carrerai carrerai and L. naiffi in Lu. hirsuta hirsuta were identified based on cytB sequencing. Bartonella spp. related to Bartonella bacilliformis in Lu. whitmani, Lu. nevesi, Lu. hirsuta hirsuta and Lu. sherlocki, and a Bartonella sp. related to Candidatus B. rondoniensis in Lu. nevesi and Lu. maranonensis were identified based on gltA gene sequencing.Conclusions/SignificanceUV, Blue LED, Mosquito Magnet and Shannon traps were more efficient than the BG-Sentinel, Green, and Red LED traps. This is the first report of L. naiffi and of two genotypes of Bartonella spp. related to B. bacilliformis and Candidatus B. rondoniensis infecting sand fly species from the Amazon region in Peru.     

中稻田三种飞虱的捕食性天敌优势种及农药对天敌的影响

对两优0923非防治的中稻田白背飞虱、灰飞虱和褐飞虱与其天敌间的关系,采用灰色关联度法、生态位分析方法,对盆拍法调查的3种飞虱与其捕食性天敌在数量、时间和空间三方面关系进行分析,对每一种天敌对应的关联度、生态位重叠指数和相似性比例等参数标准化后的密切指数相加,按照密切指数值之和大小排序,评判3种飞虱捕食性天敌优势种.并用同样方法分析常规防治田农药对飞虱捕食性天敌的影响,以期为合理施药,科学保护和利用天敌优势种提供科学依据,其结果是,非防治田白背飞虱前三位天敌是条纹蝇虎、草间小黑蛛和锥腹肖蛸;灰飞虱的是八斑球腹蛛、茶色新园蛛和锥腹肖蛸;褐飞虱的是纵条蝇狮、四点亮腹蛛和黑肩绿盲蝽.防治田白背飞虱前三位天敌是条纹蝇虎、草间小黑蛛和锥腹肖蛸;灰飞虱的是拟水狼蛛、四点亮腹蛛和草间小黑蛛;褐飞虱的是黑肩绿盲蝽、拟水狼蛛和四点亮腹蛛.盆拍法的防治田和非防治田之间3种飞虱前三位的相同天敌,白背飞虱完全相同,灰飞虱没有相同天敌,褐飞虱的是黑肩绿盲蝽和四点亮腹蛛.飞虱与天敌在时间和数量关系上,扫网法的防治田和非防治田之间3种飞虱前三位的相同天敌,白背飞虱的是锥腹肖蛸和四点亮腹蛛,灰飞虱完全相同,褐飞虱的是纵条蝇狮和条纹影虎.两种稻田的差异主要是农药杀伤了飞虱,使飞虱数量减少,并对天敌有一定杀伤力,进而影响到天敌的发生规律.非防治田的盆拍法和扫网法之间,3种飞虱前三位相同的天敌,白背飞虱的是锥腹肖蛸,灰飞虱的也是锥腹肖蛸,褐飞虱的是纵条蝇狮.防治田两调查方法结果之间,白背飞虱的是锥腹肖蛸,灰飞虱和褐飞虱前3位天敌中没有相同天敌,其差异主要是两法调查稻株的部位不同所致.     

Development of Multiplex PCR Assays for the Identification of the 33 Serotypes of Streptococcus suis

Streptococcus suis is an important zoonotic agent causing severe diseases in pigs and humans. To date, 33 serotypes of S . suis have been identified based on antigenic differences in the capsular polysaccharide. The capsular polysaccharide synthesis (cps) locus encodes proteins/enzymes that are responsible for capsular production and variation in the capsule structures are the basis of S . suis serotyping. Multiplex and/or simplex PCR assays have been developed for 15 serotypes based on serotype-specific genes in the cps gene cluster. In this study, we developed a set of multiplex PCR (mPCR) assays to identify the 33 currently known S . suis serotypes. To identify serotype-specific genes for mPCR, the entire genomes of reference strains for the 33 serotypes were sequenced using whole genome high-throughput sequencing, and the cps gene clusters from these strains were identified and compared. We developed a set of 4 mPCR assays based on the polysaccharide polymerase gene wzy, one of the serotype-specific genes. The assays can identify all serotypes except for two pairs of serotypes: 1 and 14, and 2 and 1/2, which have no serotype-specific genes between them. The first assay identifies 12 serotypes (serotypes 1 to 10, 1/2, and 14) that are the most frequently isolated from diseased pigs and patients; the second identifies 10 serotypes (serotypes 11 to 21 except 14); the third identifies the remaining 11 serotypes (serotypes 22 to 31, and 33); and the fourth identifies a new cps cluster of S . suis discovered in this study in 16 isolates that agglutinated with antisera for serotypes 29 and 21. The multiplex PCR assays developed in this study provide a rapid and specific method for molecular serotyping of S . suis .     

UncI protein can mediate ring-assembly of c-subunits of FoF1-ATP synthase in vitro

In F_oF₁-ATP synthase, multimeric c-subunits are assembled to a ring (c-ring) in the membranes that rotates as protons flow across F_o. We recently reported that assembly of c-ring of Propionigenium modestum in the membranes of Escherichia coli cells required P. modestum UncI, a product of the conserved uncI gene in the F_oF₁ operon. However, cooperation with endogenous factors in E. coli remained unclear. Here, P. modestum c-subunit was synthesized in vitro in the presence of liposomes. When c-subunit alone was synthesized, it did not form c-ring. However, when c-subunit and P. modestum UncI were synthesized together, c-ring was formed. Fusion of the two kinds of liposomes, one containing only unassembled c-subunit and the other only UncI, resulted in gradual formation of c-ring. Thus, UncI alone can mediate in vitro post-translational c-ring assembly.     

Characterization of y-type high-molecular-weight glutenins in tetraploid species of Leymus

Three y-type high-molecular-weight (HMW) glutenin gene open reading frames (ORFs), Chiy1, Chiy2, and Racy, were isolated and characterized from Leymus chinensis PI499516 and Leymus racemosus ssp. racemosus W623305. They shared an extra glutamine in the N-terminal and LAAQLPAMCRL peptides in the C-terminal with x-type HMW glutenins but had different N-terminal lengths. Like other y-type HMW glutenins, Chiy2 and Racy had 104 (or 105) amino acid (aa) residues at the N-terminal and started with EGEASR, whereas Chiy1 had 99 aa in this domain and started with QLQCER because of the deletion of EGEASR. Five other y-type glutenins, including those from Elymus ciliaris, Pseudoroegneria libanotica, and Leymus mollis, were similar to Chiy1. The ORF of Chiy2 was probably not expressed. The ORFs of both Chiy1 and Racy were expressed in bacteria. The maximum likelihood phylogenic tree based on the signal peptide and N-terminal and C-terminal aa residues revealed two clades of y-type HMW glutenins in Triticeae; the first contained Ay, By, Cy, Dy, Eey, Gy, Ky, Ry, Tay, and Uy, while the second clade contained the remaining y types, including those from Leymus. Within the second clade, HMW glutenins lacking the EGEASR peptide formed a subclade. These y-type HMW glutenins in Leymus could not be targeted to the Xm or Ns genome.     

Immunohistochemical Study of Expression of Sohlh1 and Sohlh2 in Normal Adult Human Tissues

The expression pattern of Sohlh1 (spermatogenesis and oogenesis specific basic helix-loop-helix 1) and Sohlh2 in mice has been reported in previous studies. Sohlh1 and Sohlh2 are specifically expressed in spermatogonia, prespermatogonia in male mice and oocytes of primordial and primary follicles in female mice. In this report, we studied the expression pattern of Sohlh1 and Sohlh2 in human adult tissues. Immunohistochemical staining of Sohlh1 and Sohlh2 was performed in 5 samples of normal ovaries and testes, respectively. The results revealed that Sohlh genes are not only expressed in oocytes and spermatogonia, but also in granular cells, theca cells, Sertoli cells and Leydig cells, and in smooth muscles of blood vessel walls. To further investigate the expression of Sohlh genes in other adult human tissues, we collected representative normal adult tissues developed from three embryonic germ layers. Compared with the expression in mice, Sohlhs exhibited a much more extensive expression pattern in human tissues. Sohlhs were detected in testis, ovary and epithelia developed from embryonic endoderm, ectoderm and tissues developed from embryonic mesoderm. Sohlh signals were found in spermatogonia, Sertoli cells and also Leydig cells in testis, while in ovary, the expression was mainly in oocytes of primordial and primary follicles, granular cells and theca cells of secondary follicles. Compared with Sohlh2, the expression of Sohlh1 was stronger and more extensive. Our study explored the expression of Sohlh genes in human tissues and might provide insights for functional studies of Sohlh genes.     

Fecundity as one of possible factors contributing to the dominance of the wMel genotype of Wolbachia in natural populations of Drosophila melanogaster

Alphaproteobacteria of the genus Wolbachia are common intracellular endosymbionts of a variety of insects. Their successful spread over a vast range of host taxa is often attributed to selective advantages conferred by the bacteria to infected individuals. Among the known diversity of Wolbachia pipientis infecting Drosophila melanogaster, a single genotype, wMel, within the wMel strain has been found to dominate over other genotypes world-wide. Genotyping of D. melanogaster wild populations from Ukraine reveals a relatively high frequency of the wMel genotype, although 31 % flies from an Uman’ population are infected with the rare genotype wMelCS. We demonstrate that wMelCS-infected females have lower fecundity compared to wMel-infected flies, which might be the cause of wMel prevalence in D. melanogaster populations. We report no difference in the bacterial transmission rate between these two bacterial genotypes. However, we observed an association between transmission fidelity of Wolbachia and genotype of D. melanogaster indicating that Wolbachia-host relationships in this case are more complex. Furthermore our study reveals fluctuations in Wolbachia infection rates in wMel-infected populations.     

Analysis of changes of insect-plant ratio-improvement of key-factor analysis for evaluating bottom-up effects in insect population dynamics

A revised key-factor analysis was presented for analyzing the temporal changes in the ratio of insect absolute number to plant resource. Ten data sets for 5 insect species were then analyzed. In this key-factor analysis, the key factor is defined as the factor contributing highly to between-year variation inR _r , the log rate of the inter-year change of the insect-plant ratio. The yearly change of plant resource was handled as a separate factor, expressed byr _pl , log ratio of plant resource in yearn to plant resource in yearn+1. The following was revealed: 1) In 7 of the 10 data sets examined,r _pl influenced variations ofR _r ; in particular in 3 casesr _pl was the main key factor. 2) Generation-to-generation fluctuations of absolute insect densities showed density dependence in 4 cases, while those of insect-plant ratios, in 8 cases. 3) The Royama model or a linear model, explained well the relationship between log insect-plant ratio (X _r ) andR _r and the relationship betweenX _r and log yearly change rate of absolute insect density (R _abs ). However, in the 7 cases in whichr _pl was a critical factor for variations ofR _r , with, increase ofX _r ,R _r showed a steeper, decrease around the equilibrium point (the point for whichR _r is 0) thanR _abs . This occurred becauser _pl tended to be negatively correlated withX _r . Consequently, in two casesX _r fluctuated cyclicly or chaotically although without the changes in plant resource, fluctuations ofX _r would be damped oscillations approaching equilibrium.     

Development of Heterodera glycines as Affected by Fusarium solani,the Causal Agent of Sudden Death Syndrome of Soybean

The effects of the blue form of Fusarium solani, the causal agent of sudden death syndrome (SDS), on Heterodera glycines were examined in the greenhouse. Roots of soybean cv. Coker 156 were inoculated with either H. glycines alone or F. solani + H. glycines in combination. Population levels of H. glycines were reduced 47% in the presence of F. solani. Life-stage development of H. glycines increased 3% in 30 days in the presence of F. solani. Fusarium solani colonized epidermal and cortical cells adjacent to developing juveniles of H. glycines and the nematode-induced syncytia within the soybean root tissue. At 40 days after inoculation, F. solani was isolated from 37% of the cysts in soil recovered from the F. solani + H. glycines combination treatment. Fusarium solani significantly affected H. glycines population density, life-stage development, and succeeding populations.     

Enteric bacteria of field-collected Colorado potato beetle larvae inhibit growth of the entomopathogens Photorhabdus temperata and Beauveria bassiana

The nematode Heterorhabditis marelatus fails to reproduce in the Colorado potato beetle, Leptinotarsa decemlineata, possibly due to interference from the enteric bacteria of the beetle. Specifically, the enteric bacteria inhibit the growth of Photorhabdus temperata, the enteric symbiont of the nematode, in vitro. However, previous work was based on a laboratory culture of L. decemlineata, and we wished to determine if similar bacteria were present in the field. Therefore, we cultured the enteric bacteria of fourth-instar larvae collected from the field at two locations in Maryland and Virginia. Representatives of the genera Pantoea, Enterobacter, Pseudomonas, Acinetobacter, Serratia, Stenotrophomonas, Curtobacterium, Bacillus, Lactococcus and Enterococcus were identified by sequencing of their 16S rDNA. Isolates belonging to the genera Pantoea, Enterobacter, Pseudomonas, Serratia and Bacillus inhibited the growth of P. temperata. A number of these isolates also inhibited the entomopathogenic fungus Beauveria bassiana in vitro.     

A new species and synonymy of the Neotropical Eucelatoria Townsend and redescription of Myiodoriops Townsend

The New World tropics represents the most diverse region for tachinid parasitoids (Diptera: Tachinidae), but it also contains the most narrowly defined, and possibly the most confusing, tachinid genera of any biogeographic region. This over-splitting of genera and taxonomic confusion has limited progress toward our understanding the family in this region and much work is needed to revise, redefine, and make sense of the profusion of finely split taxa. In a recent analysis of the Neotropical genus Erythromelana Townsend, two species previously assigned to this genus, Euptilodegeeria obumbrata (Wulp) and Myiodoriops marginalis Townsend were reinstated as monotypic genera. In the present study, we demonstrate that Euptilodegeeria obumbrata (Wulp), previously assigned to three different genera, represents in fact a species of the large New World genus Eucelatoria Townsend, in which females possess a sharp piercer for oviposition. We also show that the species Eucelatoria carinata (Townsend) belongs to the same species group as Eucelatoria obumbrata, which we here define and characterize as the Eucelatoria obumbrata species group. Additionally, we describe Eucelatoria flava sp. n. as a new species within the Eucelatoria obumbrata species group. Finally, we redescribe the genus Myiodoriops Townsend and the single species Myiodoriops marginalis Townsend.     

Effect of Eradication of Helicobacter pylori on Expression Levels of FHIT,IL-8 and P73 in Gastric Mucosa of First-Degree Relatives of Gastric Cancer Patients

Objectives

Helicobacter pylori (H. pylori) infection plays an important role in the carcinogenesis and development of gastric cancer. Eradication of H. pylori can effectively reduce the risk of gastric cancer, but the underlying mechanisms are not fully understood. This study aimed to investigate the effect of eradication of H. pylori on the expression levels of FHIT, IL-8 and P73 in the gastric mucosa of first-degree relatives of gastric cancer patients.

Methods

One hundred and thirty-two patients with functional dyspepsia having first-degree relatives with gastric cancer were prospectively recruited in this study. Nine patients presented with H. pylori infection and family histories of gastric cancer, 61 with H. pylori infection and without family histories of gastric cancer, 6 without H. pylori infection and with family histories of gastric cancer, and 56 without H. pylori infection and family histories of gastric cancer. The protein and mRNA expression levels of FHIT, IL-8 and P73 in gastric mucosa of the subjects were detected by immunohistochemical staining and polymerase chain reaction, respectively.

Results

Compared with the patients without H. pylori infection and family histories of gastric cancer, both the protein and mRNA levels of FIHT significantly decreased in patients with H. pylori infection and/or family histories of gastric cancer, and both the protein and mRNA levels of IL-8 significantly increased. After eradication of H. pylori, both the protein and mRNA levels of FHIT were significantly higher, and both the protein and mRNA levels of IL-8 were significantly lower. However, H. pylori infection and family histories of gastric cancer had no major effect on P73 expression.

Conclusions

Down-regulation of FHIT and up-regulation of IL-8 may be involved in the pathogenesis of H. pylori infection in the first-degree relatives of gastric cancer patients.