IL-31 Associated with Coronary Artery Lesion Formation in Kawasaki Disease

Background

Kawasaki disease (KD) is known to be associated with T help (Th) 2 reaction and subsequently allergic diseases. Interleukin-31 (IL-31) has also been reported to be involved in Th2 mediated diseases such as allergic diseases. However, the role of IL-31 in KD has not been previously reported. The aim of this study is to investigate whether IL-31 is associated with KD and its clinical outcome.

Material

A total of 78 KD patients who met the criteria of KD were enrolled in this study as well as 20 age-matched controls. Plasma samples were conducted to measure IL-31 before intravenous immunoglobulin (IVIG) treatment (KD1), within 3 days after IVIG treatment (KD2) and at least 3 weeks after IVIG treatment (KD3) by utilizing enzyme-linked immunosorbent assay (ELISA).

Result

Our findings showed that IL-31 expression was higher in KD patients after IVIG treatment significantly (KD2>KD1: 1265.0±199.3 vs. 840.2±152.5 pg/ml, p<0.0001). Further analysis revealed that IL-31 level was significantly higher in KD patients with coronary artery lesion (CAL) (656.6±139.5 vs. 1373.0±422.0 pg/ml, p = 0.04) before IVIG treatment (KD1). There were no significant differences between the IVIG resistance and IVIG responsiveness groups.

Conclusion

IL-31 was increased after IVIG treatment in patients with KD and was significantly associated with CAL formation. The results from this study may help to identify a novel risk factor for predicting KD and CAL formation.     

Immunologic identification of the aromatase enzyme system in human endometrium

Confluent human endometrial stromal cells were cultured in medium with no hormone or supplemented with medroxyprogesterone acetate (MPA), estradiol (E2), and porcine relaxin (RLX) for 5 days. These stromal cells were then labeled with [35S]methionine for 3 h. The radioactive proteins in the particulate fraction of cell homogenate were extracted by detergent and incubated with antisera to purified placental aromatase cytochrome P-450 (P-450arom) and NADPH-cytochrome P-450 reductase to isolate the radio-labeled aromatase enzyme components. Analysis of the radio-labeled protein, isolated by antibody to the cytochrome P-450arom from different preparations (P45FBIII or R-8-2) showed a major band at molecular weight 54k on SDS polyacrylamide gel electrophoresis (SDS-PAGE). The intensity of 54k band was stronger in hormone treated stromal cells than that of control in parallel with the increase of aromatase activity. The radio-labeled protein isolated by anti-NADPH cytochrome P-450 reductase, REDFBIV, showed a major band at the molecular weight 73k on SDS-PAGE with comparable intensity in control and hormone treated samples. Thus, the apparent molecular weights of endometrial cytochrome P-450arom and cytochrome P-450 reductase were identical to placental aromatase enzyme system. When a secretory endometrium and a decidua were labeled with [35S]methionine, the cytochrome P-450arom was detected only in the decidua. NADPH cytochrome P-450 reductase was detected both in the endometrium and the decidua. These results show that antisera to placental aromatase enzyme system cross reacts with the endometrial aromatase enzyme components. The synthesis of cytochrome P-450arom was stimulated by MPA, E2 and RLX while the synthesis of the NADPH-cytochrome P-450 reductase aromatase component was not affected by the hormone.     

Focus on Weed Control: The Impact of Herbicide-Resistant Rice Technology on Phenotypic Diversity and Population Structure of United States Weedy Rice

The use of herbicide-resistant (HR) Clearfield rice (Oryza sativa) to control weedy rice has increased in the past 12 years to constitute about 60% of rice acreage in Arkansas, where most U.S. rice is grown. To assess the impact of HR cultivated rice on the herbicide resistance and population structure of weedy rice, weedy samples were collected from commercial fields with a history of Clearfield rice. Panicles from each weedy type were harvested and tested for resistance to imazethapyr. The majority of plants sampled had at least 20% resistant offspring. These resistant weeds were 97 to 199 cm tall and initiated flowering from 78 to 128 d, generally later than recorded for accessions collected prior to the widespread use of Clearfield rice (i.e. historical accessions). Whereas the majority (70%) of historical accessions had straw-colored hulls, only 30% of contemporary HR weedy rice had straw-colored hulls. Analysis of genotyping-by-sequencing data showed that HR weeds were not genetically structured according to hull color, whereas historical weedy rice was separated into straw-hull and black-hull populations. A significant portion of the local rice crop genome was introgressed into HR weedy rice, which was rare in historical weedy accessions. Admixture analyses showed that HR weeds tend to possess crop haplotypes in the portion of chromosome 2 containing the ACETOLACTATE SYNTHASE gene, which confers herbicide resistance to Clearfield rice. Thus, U.S. HR weedy rice is a distinct population relative to historical weedy rice and shows modifications in morphology and phenology that are relevant to weed management.Weedy rice (Oryza sativa), a conspecific weed of cultivated rice, is a global threat to rice production (Delouche et al., 2007). Classified as the same species as cultivated rice, it is highly competitive (Diarra et al., 1985; Pantone and Baker, 1991; Burgos et al., 2006), difficult to control without damaging cultivated rice, and can cause almost total crop failure (Diarra et al., 1985). The competition of cultivated rice with weedy rice can lead to yield losses from less than 5% to 100% (Kwon et al., 1991; Watanabe et al., 2000; Chen et al., 2004; Ottis et al., 2005; Shivrain et al., 2009b). Besides being difficult to control, weedy rice persists in rice fields because of key weedy traits, including variable emergence (Shivrain et al., 2009b), high degree of seed shattering (Eleftherohorinos, et al., 2002; Thurber et al., 2010), high diversity in seed dormancy (Do Lago, 1982; Noldin, 1995; Vidotto and Ferrero, 2000; Burgos et al., 2011; Tseng et al., 2013), and its seed longevity in soil (Goss and Brown, 1939). Weedy rice is a problem mainly in regions with large farm sizes where direct-seeded rice culture is practiced (Delouche et al., 2007). It is not a major problem in transplanted rice culture, where roguing weeds is possible and hand labor is available. The severity of the problem has increased in recent decades because of the significant shift to direct seeding from transplanting (Pandey and Velasco, 2002; Rao et al., 2007; Chauhan et al., 2013), which is driven by water scarcity (Kummu et al., 2010; Turral et al., 2011), increasing labor costs, and migration of labor to urban areas (Grimm et al., 2008).The herbicide-resistant (HR) Clearfield rice technology (Croughan, 2003) provides an option to control weedy rice in rice using imidazolinone herbicides, in particular, imazethapyr. Imidazolinones belong to group 2 herbicides, also known as ACETOLACTATE SYNTHASE (ALS) inhibitors. Examples of herbicides in this group are imazamox, imazapic, imazaquin, and imazethapyr. Developed through mutagenesis of the ALS locus (Croughan, 1998), Clearfield rice was first commercialized in 2002 in the southern U.S. rice belt (Tan et al., 2005). Low levels of natural hybridization are known to occur between the crop and weedy rice. Gene flow generally ranges from 0.003% to 0.25% (Noldin et al., 2002; Song et al., 2003; Messeguer et al., 2004; Gealy, 2005; Shivrain et al., 2007, 2008). After the adoption of Clearfield technology, resistant weedy outcrosses were soon detected in commercial fields (Fig. 1), generally after two cropping seasons of Clearfield rice, where escaped weedy rice was able to produce seed (Zhang et al., 2006; Burgos et al., 2007, 2008). Similar observations have been reported outside the United States, in other regions adopting the technology (Gressel and Valverde, 2009; Busconi et al., 2012).Open in a separate windowFigure 1.Suspected herbicide-resistant weedy rice in a rice field previously planted with Clearfield rice along the Mississippi River Delta in Arkansas. More than 10 morphotypes of weedy rice were observed in this field, with different maturity periods. In the foreground is a typical weedy rice with pale green leaves; the rice cultivar has dark green leaves. The inset shows a weedy morphotype that matured earlier than cultivated rice.Despite this complication, the adoption of Clearfield rice technology is increasing, albeit at a slower pace than that of glyphosate-resistant crops. After a decade of commercialization, 57% of the rice area in Arkansas was planted with Clearfield rice cultivars in 2013 (J. Hardke, personal communication). Clearfield technology has been very successful at controlling weedy rice, and polls among rice growers suggest that farmers have kept the problem of HR weeds in check by following the recommended stewardship practices (Burgos et al., 2008). The most notable of these are (1) implementation of herbicide programs that incorporate all possible modes of action available for rice production; (2) ensuring maximum efficacy of the herbicides used; (3) preventing seed production from escaped weedy rice, remnant weedy rice after crop harvest, or volunteer rice and weedy rice in the next crop cycle; (4) rotating Clearfield rice with other crops to break the weedy rice cycle; and (5) practicing zero tillage to avoid burying HR weedy rice seed (Burgos et al., 2008).Clearfield rice has gained a foothold in Asia, where rice cultivation originated (Londo and Schaal, 2007; Zong et al., 2007). Clearfield rice received government support for commercialization in Malaysia in 2010 (Azmi et al., 2012) because of the severity of the weedy rice problem there. Dramatic increases in rice yields (from 3.5 to 7 metric tons ha⁻¹) were reported in Malaysia where Clearfield rice was planted (Sudianto et al., 2013). However, the risk of gene flow and evolution of resistant weedy rice populations is high in the tropics, where up to three rice crops are planted each year, and freezing temperatures, which would reduce the density of volunteer plants, do not occur.In the United States, where Clearfield technology originated and has been used for the longest time, the interaction between HR cultivated rice and weedy rice is not yet fully understood. Two main populations of weedy rice are known to occur in the southern United States and can be found in the same cultivated rice fields. These populations are genetically differentiated, are largely distinct at the phenotypic level, and have separate evolutionary origins (Reagon et al., 2010). One group tends to have straw-colored hulls and is referred to as the SH population; a second group tends to have black-colored hulls and awns and is referred to as the BHA population (Reagon et al., 2010). Genomic evidence suggests that both groups descended from cultivated ancestors but not from the tropical japonica subgroup varieties that are grown commercially in the United States. Instead, the SH group evolved from indica, a subgroup of rice commonly grown in the lowland tropics, and the BHA group descended from aus, a related cultivated subgroup typically grown in Bangladesh and the West Bengal region (Reagon et al., 2010). Weed-weed and weed-crop hybrids are also known to occur, but prior to Clearfield commercialization, these hybrids had occurred at low frequency (Reagon et al., 2010; Gealy et al., 2012). With the advent and increased adoption of Clearfield cultivars, the impact on U.S. weedy rice population structure and the prevalence of the SH and BHA groups are unknown.Efforts to predict the possible consequences of HR or genetically modified rice on weedy rice have been a subject of discussion for many years. Both weedy rice and cultivated rice are primarily self-fertilizing, but, as mentioned above, low levels of gene flow are known to occur. Additional environmental and intrinsic genetic factors can act as prezygotic and postzygotic mating barriers between cultivated and weedy rice and influence the possibility and levels of gene flow between these groups (Craig et al., 2014; Thurber et al., 2014). However, once gene flow occurs between cultivated and weedy rice, and if the resulting hybrids are favored by selection, the resulting morphological, genetic, and physiological changes in weedy rice populations can alter the way that weedy rice evolves and competes. For example, herbicide-resistant weed outcrosses in an experimental field have been observed to be morphologically diverse (Shivrain et al., 2006), with some individuals carrying major weedy traits and well adapted to rice agriculture. Such weedy plants could be more problematic than their normal weedy counterparts. Thus, introgression of crop genes into weedy populations has the potential to change the population dynamic, genetic structure, and morphological profile of weedy plants. This, in turn, must alter our crop management practices. To increase our understanding of the impact of HR rice on the evolution of weedy rice, in this article we aim to (1) assess the frequency of herbicide resistance in weedy rice in southern U.S. rice fields with a history of Clearfield use; (2) characterize the weedy attributes of resistant populations; and (3) determine the genetic origins of herbicide-resistant weeds in U.S. fields.     

Antimicrobial Activity of Antrodia camphorata Extracts against Oral Bacteria

Antrodia camphorata (A. camphorata) is a unique, endemic and extremely rare mushroom species native to Taiwan, and both crude extracts of and purified chemical compounds from A. camphorata have been reported to have a variety of significant beneficial effects, such as anti-tumor and anti-inflammatory activity. However, reports on the effects of A. camphorata against dental pathogens have been limited. Oral health is now recognized as important for overall general health, including conditions such as dental caries, periodontal disease and rheumatoid arthritis. Streptococcus mutans (S. mutans) and Porphyromonas gingivalis (P. gingivalis) are the most common bacteria associated with dental plaque and periodontopathic diseases, respectively. Thus, our study examined the ability of five various crude extracts of A. camphorata to inhibit the growth of dental bacteria and anti-adherence in vitro. Among the extracts, the ethanol, ethyl acetate and chloroform extracts exhibited the lowest MICs against P. gingivalis and S. mutans (MIC = 4∼16 µg/mL). The MIC of the aqueous extract was greater than 2048 µg/mL against both P. gingivalis and S. mutans. In vitro adherence of S. mutans was significantly inhibited by the addition of either the ethyl acetate extract or chloroform extract (MIC = 16∼24 µg/mL), while the ethanol extract (MIC = 32∼64 µg/mL) exhibited moderate inhibitory activity. Based on the result of this study, the ethyl acetate and chloroform extracts of A. camphorata may be good candidates for oral hygiene agents to control dental caries and periodontopathic conditions.     

A Single Divergent Exon Inhibits Ankyrin-B Association with the Plasma Membrane

Vertebrate ankyrin-B and ankyrin-G exhibit divergent subcellular localization and function despite their high sequence and structural similarity and common origin from a single ancestral gene at the onset of chordate evolution. Previous studies of ankyrin family diversity have focused on the C-terminal regulatory domain. Here, we identify an ankyrin-B-specific linker peptide connecting the ankyrin repeat domain to the ZU5₂-UPA module that inhibits binding of ankyrin-B to membrane protein partners E-cadherin and neurofascin 186 and prevents association of ankyrin-B with epithelial lateral membranes as well as neuronal plasma membranes. The residues of the ankyrin-B linker required for autoinhibition are encoded by a small exon that is highly divergent between ankyrin family members but conserved in the ankyrin-B lineage. We show that the ankyrin-B linker suppresses activity of the ANK repeat domain through an intramolecular interaction, likely with a groove on the surface of the ANK repeat solenoid, thereby regulating the affinities between ankyrin-B and its binding partners. These results provide a simple evolutionary explanation for how ankyrin-B and ankyrin-G have acquired striking differences in their plasma membrane association while maintaining overall high levels of sequence similarity.     

Affinity purification of Candida albicans CaCdc4-associated proteins reveals the presence of novel proteins involved in morphogenesis

Candida albicans CDC4 is nonessential and plays a role in suppressing filamentous growth, in contrast to its evolutionary counterparts involved in the G1-S transition of the cell cycle. Genetic epistasis analysis has indicated that proteins besides Sol1 are targets of C. albicans Cdc4. Moreover, no formal evidence suggests that C. albicans Cdc4 functions through the ubiquitin E3 ligase of the Skp1-Cul1/Cdc53-F-box complex. To elucidate the role of C. albicans CDC4, C. albicans Cdc4-associated proteins were sought by affinity purification. A 6×His epitope-tagged C. albicans Cdc4 expressed from Escherichia coli was used in affinity purifications with the cell lysate of C. albicans cdc4 homozygous null mutant. Candida albicans Cdc4 and its associated proteins were resolved by SDS-PAGE and visualized by silver staining. The candidate proteins were recovered and trypsin-digested to generate MALDI-TOF spectra profiles, which were used to search against those of known proteins in the database to reveal their identities. Two out of four proteins encoded by GPH1 and THR1 genes were further verified to interact with C. albicans Cdc4 using a yeast two-hybrid assay. We conclude that in vitro affinity purification using C. albicans Cdc4 generated from E. coli as the bait and proteins from cell lysate of C. albicans cdc4 homozygous null mutant as a source of prey permit the identification of novel proteins that physically interact and functionally associate with C. albicans Cdc4.     

Mitotic chromosomes of species in the subgenusDrosophila (Diptera:Drosophilidae)

The karyotypes of 17 species in the subgenusDrosophila are compared according to their taxonomical relationships. Although closely related species often possess similar karyotypes, the karyotypes diverge considerably within the subgenus. Thus extensive chromosome rearrangements did occur during the speciation. Species with higher chromosome numbers do not necessarily have higher average of total chromosome length per cell.     

Phototropins and Their LOV Domains：Versatile Plant Blue-Light Receptors

The phototropins phot1 and phot2 are plant blue-light receptors that mediate phototropism, chloroplast movements, stomatal opening, leaf expansion, the rapid Inhibition of hypocotyl growth in etiolated seedlings, and possibly solar tracking by leaves in those species in which It occurs. The phototroplns are plasma membrane-associated hydrophilic proteins with two chromophore domains （designated LOV1 and LOV2 for their resemblance to domains In other signaling proteins that detect light, oxygen, or voltage） in their Nterminal half and a classic serine/threonlne kinase domain in their C-terminal half. Both chromophore domains bind flavin mononucleotide （FMN） and both undergo light-activated formation of a covalent bond between a nearby cystelne and the C（4a） carbon of the FMN to form the signaling state. LOV2-cystelnyl adduct formation leads to the release downstream of a tightly bound amphlpathlc α-helix, a step required for activation of the klnase function. This cysteinyl adduct then slowly decays over a matter of seconds or minutes to return the photoreceptor chromophore modules to their ground state. Functional LOV2 is required for light-activated phosphorylation and for various blue-light responses mediated by the phototroplns. The function of LOV1 is still unknown, although It may serve to modulate the signal generated by LOV2. The LOV domain Is an ancient chromophore module found In a wide range of otherwise unrelated proteins In fungi and prokaryotes, the latter Including cyanobacterla, eubacterla, and archaea. Further general reviews on the phototropins are those by Celaya and Liscum （2005） and Christie and Briggs （2005）.     

Risk Factors for Progressive Visual Field Loss in Primary Angle-Closure Glaucoma: A Retrospective Cohort Study

Purpose

To investigate risk factors associated with progressive visual field (VF) loss in primary angle closure glaucoma (PACG).

Methods

We retrospectively reviewed medical record of PACG patients who had ≥5 reliable VF examinations (central 24-2 threshold test, Humphrey Field Analyzer) and ≥2 years of follow-up. Each VF was scored using Collaborative Initial Glaucoma Treatment Study system. Progression was defined if 3 consecutive follow-up VF tests had an increased score of ≥3 above the mean of the first 2 VF scores. Factors associated with VF progression were evaluated by Cox proportional hazards models.

Results

A total of 89 eyes from 89 patients (mean age, 69.8 ± 7.9 years), who received a mean of 6.9 ± 2.3 VF tests (mean deviation at initial, -8.1 ± 4.4 dB) with a mean follow-up of 63.9 ± 23.9 months were included. VF progression was detected in 9 eyes (10%). The axial length (AL), anterior chamber depth, and intraocular pressure (IOP) in patients with and without progression were 22.5 ± 0.6 and 23.1 ± 0.9 mm, 2.5 ± 0.3 and 2.5 ± 0.3 mm, 14.8 ± 2.4 and 14.3 ± 2.3 mm Hg, respectively. AL was the only factor associated with progression in both Cox proportional hazards univariate (p = 0.031) and multivariate models (p = 0.023).

Conclusion

When taking into account age, IOP, follow-up period, and number of VF tests, a shorter AL is the only factor associated with VF progression in this cohort of Chinese patients with PACG. Further studies are warranted to verify the role of AL in progressive VF loss in PACG.     

Isolation and characterization of yeast nicotinamide adenine dinucleotide kinase.

NAD+ kinase (ATP:NAD+ 2'-phosphotransferase, EC 2.7.1.23) from yeast has been purified utilizing ion-exchange and NAD+-agarose affinity chromatography to give a 2100-fold purification. The apparent homogeneity of the enzyme preparation was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and analytical ultracentrifugation. The enzyme has a subunit molecular weight of 31,000, and a native molecular weight of 124,000, and is, thus, probably a tetramer. The single form of the enzyme has an apparent isoelectric point of 5.85. Initial velocity studies in the forward direction with both substrates gave intersecting Lineweaver-Burk plots, and this suggests a sequential mechanism in which both substrates are bound before products are released. Replots of these data were linear and gave Km values for NAD+ and ATP of 0.68 mM and 2.3 mM, respectively.