The observed elevation in TRAP1 protein abundance requires further validation

to determine whether enhanced TRAP content may limit cell damage and regulate cell repair for restoration or apoptosis after elevated stress response [60]. The functional role of PARK7 in skeletal muscle is still unknown, but PARK7 knockout mice show a reduced mitochondrial ACO2 activity and enhanced mitochondrial glutathione peroxidase activity, which suggests a deficient mitochondrial H2O2 scavenging function [61] and [62]. We report that ACO2 abundance is increased in myotubes from T2D patients, while PARK7 protein level is reduced. Whether PS-341 mouse there is a direct connection between these proteins in metabolic pathways and disease predisposition requires further investigation. However, differential protein profiles of chaperones in myotubes derived from T2D patients supports the the growing idea that disturbances in the protein maintanance system may cause impaired mitochondrial quality control system and thereafter a fundamental disturbance of cellular

metabolic activity [55]. A comparison of the proteome of myotubes derived from NGT versus T2D patients revealed that several proteins involved in mRNA processing, regulation and transcription are altered. This finding advocates the idea that T2D imparts a disease-related inhibition of basic cellular functions in skeletal muscle. For example, KHSRP, a key mediator of mRNA decay known to promote the biogenesis of a subset of microRNAs [63], was more abundant Cyclin-dependent kinase 3 in myotubes from E7080 research buy T2D patients. KHSRP is phosphorylated by p38MAPK and Akt in the regulation of the mRNA degradation pathway [64] and turnover of myogenic mRNA [65]. Therefore, while KHSRP is more abundant in T2D myotubes, its role and function requires further study in relation to insulin

resistance. Proteome analysis also revealed that myotubes derived from T2D patients possess higher levels of the DNA repair proteins, XRCC5, and RECQL. The function of these proteins in metabolism and T2D is still elusive. Whether an increased DNA repair activity may reflect an enhanced oxidative stress caused by increased ROS and/or reduced oxidative defense remains to be determined. The differential proteome signatures of myotubes derived from people with T2D versus NGT offers new insights into causes of T2D, highlighting pathways involving disturbances in energy metabolism, oxidative stress response, protein dynamics and gene regulation. The analysis presented here demonstrates a clear disturbance of the protein signature in skeletal muscle myotubes derived from T2D patients compared to NGT subjects. Our results reveal that metabolic impairments, reductions in GSH concentration, and differences in the protein profile are retained in cultured differentiated myotubes from T2D subjects. Thus, our findings emphasize that an intrinsic proteome exists, directed by either epigenetic or genetic factors, in skeletal muscle from T2D patients.

αvβ8−/− CD103+ DCs MK-2206 mw also

showed reduced production of inflammatory cytokines compared with CD103− DCs 6 ( Supplementary Figure 3), indicating that reduced TGF-β activation by αvβ8 does not result in an overt proinflammatory phenotype in CD103+ intestinal DCs. Data presented previously were obtained using intestinal DC subsets isolated from mLN, which include DCs draining from the small and large intestine. To determine whether CD103+ DCs present within intestinal tissues show a similar reliance on integrin αvβ8-mediated TGF-β activation to induce Foxp3+ iTregs, we first analyzed expression of β8 integrin on DCs isolated from small and large intestinal lamina propria. Similar to mLN DC subsets, CD103+ DCs from both the small and large intestine expressed high levels of β8 integrin (Figure 5A). Additionally, CD103+ DCs from both small and large intestine supported enhanced Foxp3+ iTreg induction versus CD103− DCs, which was completely reliant on expression of integrin αvβ8 ( Figure 5B). As

observed for mLN, iTreg induction in αvβ8−/− CD103+ DCs from small or large intestine was rescued by addition of active TGF-β ( Figure 5C). Interestingly, we observed slightly elevated expression of β8 on CD103− DCs from BMS-907351 research buy large intestinal lamina propria versus CD103− DCs from small intestine ( Figure 5A), mLN, and spleen (data not shown). However, such expression did not translate into an enhanced ability to induce iTreg, indicating a potentially novel role for β8 expression on CD103− DCs from the large intestine ( Figure 5B). Taken together, these data show that increased αvβ8-mediated TGF-β activation by intestinal CD103+ DCs is critical for their enhanced Edoxaban ability to induce Foxp3+ iTregs ex vivo. We next sought to determine whether integrin αvβ8 expression by intestinal

DCs supported enhanced Foxp3+ iTreg induction in vivo. To this end, we adoptively transferred ovalbumin antigen-specific CD4+ OT-II T cells into control or Itgb8 (CD11c-Cre) mice and supplemented drinking water with ovalbumin. T cells were isolated from OT-II/Rag−/− mice, which lack endogenous Foxp3+ Tregs. Previous experiments using this method have shown that Foxp3+ iTreg induction is promoted specifically in the mLN, at least in part via the enhanced ability of intestinal CD103+ DCs to promote iTreg induction. 6 and 7 In control mice, we observed ∼5% induction of Foxp3+ iTregs arising from adoptively transferred OT-II T cells specifically within the mLN (Figure 6A). This induced population was not observed in the spleen or in mice not fed ovalbumin (data not shown).

Access to instant test results would enable the growers to implement timely orchard management practices. The commonly utilized bacterial gene used for laboratory based diagnostics of

Las is a fragment of the 16S rDNA gene (Li et al., 2006). We have developed LAMP primer sequences for the phage related region of the Las genome. If Las positives are found, the crude extracts used in LAMP assays can be re-evaluated in diagnostic laboratories by qPCR for the 16S rDNA region. Utilization of two different genomic regions will be beneficial in detecting potential contaminations. The LAMP assay developed here is about 100 times more sensitive than standard qPCR Trametinib molecular weight technique and hence likely to detect the bacterium in low-titer situations (Fig. 3). While testing a large number of plant DNA samples of different varieties, we observed that it was easier to discriminate between weak positives Selleck Pifithrin�� and negatives in LAMP assays rather than in qPCR assays where samples with Ct values of 34 or above are generally considered inconclusive. Very high levels of sensitivity of LAMP reactions have been reported in many other systems. In the filarial parasite

Loa loa associated with a tropical human disease Loiasis, it has been shown that LAMP assay can detect 0.5 ag of the worm genomic DNA; compared to the qPCR test that detects 0.1 pg, LAMP was considered 200,000 times more sensitive ( Fernandez-Soto et al., 2014). In our analysis, the increased sensitivity observed in LAMP compared to qPCR may be because Urease of two reasons: a) qPCR is generally conducted in a duplex format to detect plant gene (‘Cox’) or psyllid gene (‘wingless’) in addition to the bacterial gene (16S rDNA). When the bacterial titer is low, amplification of the internal control genes may deplete the reagents required for the amplification of the bacterial target gene and this may negatively affect the Ct value calculated in qPCR assays; b) when target concentration is low, the PCR inhibitors present in the extract may negatively affect qPCR Ct values. Such an effect seems to be less of a problem with LAMP reactions. When the qPCR

assays were conducted for only Las (without housekeeping gene) and the bacterial titers were low, the sensitivity of qPCR appeared to be lower than LAMP. The Liberibacter genomic region chosen by our study seems to be specific to Las and is not reported from other bacteria. Psyllids are known to harbor several endosymbiotic bacteria like Wolbachia (alpha proteobacterial group) and Candidatus Carsonella (gamma proteobacteria) ( Saha et al., 2012). The LAMP reaction was always negative with known Las-free ACP and B. cockerelli indicating that the primers did not amplify DNA from the endosymbiont populations. If the LAMP technology is extended in the future to test plant root samples that are reported to have the bacterium earlier than the canopy ( Johnson et al.

“
“The authors would like to correct the omission of a reference in their discussion of the genetic interaction between Sdo1 and Tif6 in yeast. This work was published by Menne TF, Goyenechea B, Sanchez-Puig N, Wong, C. C., Tonkin, L. M., Ancliff, P. J., Brost, R. L., Costanzo, M., Boone, C., Warren, A. J.. The Shwachman–Bodian–Diamond GSK1120212 concentration syndrome protein mediates translational activation of ribosomes in yeast. Nat Genet. 2007;39:486–495. “
“The authors regret that the above article contained a minor error on page 2: the twenty first line of the right-hand column should read, “In normal red cells the ratio between reduced and oxidized glutathione (GSSG)

is usually about 100:1” as opposed to “In normal red cells the ratio between oxidized and reduced glutathione (GSSH) is 100:1”. “
“Sickle cell disease (SCD), the most common inherited red blood cell (RBC) disorder, affects individuals of African, Mediterranean, and Asian descent and manifests as haemolysis and vaso-occlusion [1]. Patients experience a

spectrum of disease symptoms and complications, including periods of acute pain (vaso-occlusive episodes [VOE]), chronic pain, multi-organ injury, reduced quality of life, and a shortened lifespan [1] and [2]. Worldwide it is estimated that over 200,000 children affected with SCD are born every year, primarily in selleck chemical sub-Saharan Africa (180,000 births per year) [3] and [4]. Approximately 2000 children in the US [5] are born with SCD each year, with a disease incidence of 1 in 2474 live births (newborn screening data 1990–1999) [6]; the estimated US prevalence ranges from 70,000–140,000 [7] and [8]. Among individuals with the homozygous sickle haemoglobin mutation (HbSS) living in first-world countries, the estimated mean life expectancy Tacrolimus (FK506) is 39 years [8], which has improved significantly over the last few decades. Increased overall survival of paediatric patients with SCD [9] (Fig. 1) can be attributed to the landmark Prophylactic Penicillin Study (PROPS; 1986), [10] which demonstrated that the use of prophylactic

penicillin could prevent life-threatening infections in affected children. Thus, universal newborn screening became standard practice in the US in the late 1990s and in the United Kingdom in the early 2000′s [10], [11] and [12], enabling early diagnosis and patient management. The introduction of a pneumococcal conjugate vaccine also significantly contributes to decreased SCD mortality in children younger than 10 years of age [12] and [13]. However, in low-resource countries, more than 50% of children younger than 5 years of age die due to complications of SCD [14]. Because more than 98% of children with milder forms of SCD in high-resource countries are living into adulthood, SCD is now a chronic condition requiring comprehensive, life-long management [9].

Animals were subjected to restraint stress by placing them in a metal restrainer (17 × 6 × 5 cm) for 15 min, followed by decapitation and blood collection for corticosterone and ACTH measurements. The groups were as follows: Wistar basal (n = 5), Wistar restraint (n = 6), WAR basal (n = 4) http://www.selleckchem.com/products/pci-32765.html and WAR restraint (n = 5). To determine the adrenal responsiveness to ACTH 24 h before the experiments, rats were anesthetized with 2,2,2-tribromoethanol (25 mg/100 g bw. i.p., Aldrich, Milwaukee, WI, USA); a catheter was inserted into the right external jugular vein and advanced to the right atrium (Harms and Ojeda, 1974) for i.v. drug administration. On the day of the experiment, rats were pretreated

with dexamethasone (100 μg/100 g subcutaneously) and 2 h later they received an i.v. injection of ACTH (Synacthène, Novartis—8 ng/rat) or vehicle (0.9% NaCl). Trunk blood samples for plasma corticosterone determination were collected by decapitation 15 min after the injection. Groups: Wistar vehicle (n = 4), Wistar ACTH (n = 5), WAR vehicle (n = 4) and WAR ACTH (n = 5). Rucaparib datasheet Plasma hormone levels were determined by specific radioimmunoassay, as previously described (Elias et al., 2002). The assay sensitivity was 0.4 μg/dl for corticosterone and 16 pg/ml for ACTH. The intra- and inter-assay coefficients of variation were 4% and 8% for corticosterone and 4.3% and 16% for ACTH, respectively. All samples from a single experiment were assayed

in duplicate in the same assay. Data were expressed as means ± SEM. Two-way ANOVA was used to analyze the data obtained from experiments on circadian rhythm variation, restraint stress

and exogenous ACTH stimulation. To analyze adrenal gland weight, adrenal medulla area and adrenal cortex layers, the Mann–Whitney test was used. Significance was established at p < 0.05. Eduardo HL Umeoka: Experimental procedures, data analysis and article preparation. Sérgio Britto Garcia: Histopathology and morphometry analysis of adrenal gland. José Antunes-Rodrigues, Lucila LK Elias and Norberto Garcia-Cairasco: Buspirone HCl Experimental design, advice on execution of experimental protocols/methods and article preparation. This project is supported by FAPESP, FAPESP-Cinapce, CAPES, PROEX-Physiology and PROEX-Neurology, FAEPA. JAR, LLEK and NGC are recipients of CNPq-Brazil research fellowship. We wish to thank all members of the Neuroendocrinology Laboratory and Neurophysiology and Experimental Neuroethology Laboratory, especially Rodrigo Rorato, Mauricio Benedetti, Olagide Castro, Victor Santos Rodrigues and Simone Marroni. The authors also wish to thank Maria Valci Silva, Marina Holanda and Rosangela Orlandin Lopes for their technical support. “
“In the above article, Fig. 1 and the legend for Fig. 1 appeared incorrectly. The correct Fig. 1 and legend for Fig. 1 are included here. “
“The five factor model is one of the most extensively applied models of personality currently in use.

huxleyi

requires high P concentrations relative to N: this is what we observed in our study. E. huxleyi is a cosmopolitan species, widely distributed in both oceanic and coastal waters ( Balch et al. 1991). E. huxleyi may have an unusually high affinity for P uptake and can also use alkaline phosphatase to access dissolved organic P sources ( Riegmann et al. 2000). Here, the main environmental drivers of the phytoplankton communities were wind speed/direction NVP-BGJ398 datasheet and nutrient ratios. We propose that wind speed has a strong impact on this coastal ecosystem based on the principle that in a shallow water column (i.e. 20 m), the wind speed is proportional to the bottom stress on the ocean floor and then to the resuspension of sediment and associated nutrients. Over the course of twelve months, this study demonstrated selleckchem a typical austral-seasonal pattern in water temperature, accompanied by a similar annual cycle in phytoplankton. The main species contributing to the Chl a signal

were Pyramimonas spp., Hemiselmis sp., Gyrodinium sp., Heterocapsa rotunda, Cylindrotheca closterium, Chaetoceros spp., Chrysochromulina spp. and Emiliania huxleyi. The different phytoplankton groups showed shifts in species dominance between summer and winter, with a dominance of chlorophytes during six months of the year. It became apparent that wind speed and direction played an important role in setting the environmental conditions off Port Stanvac and subsequently on the distribution and abundance of phytoplankton species in this coastal area. In summary, our results show that in the coastal waters of the GSV, phytoplankton communities are affected by wind conditions and by changing nutrient learn more levels on a seasonal basis, which is typical of coastal environments. Nutrient enrichment of coastal waters is generally the main factor driving the succession and composition of phytoplankton communities, and further work is now needed to identify the sources of nutrients in this region, where river run-off is limited and evaporation is high relative to precipitation. This is particularly relevant in the light

of environmental studies on the impact of the Adelaide Desalination Plant, which became fully operational in early December 2012. The authors acknowledge the financial support of the National Centre of Excellence in Desalination Australia which is funded by the Australian Government through the Water for the Future initiative. The authors are grateful to Shaun Byrnes, John Luick and Charles James for their help with the sampling and processing of the oceanographic data. We would also like to thank Lorenzo Andreacchio, Satish Dogra and the crew of the r/v ‘Ngerin’ for their help during sampling trips. “
“The Chinese mitten crab Eriocheir sinensis is a well-known non-native species introduced in ballast tanks to European waters almost one hundred years ago ( Peters & Panning, 1939, Gollasch 2006).

The dashed line in Fig. 2 and Fig. 3 represents the recorded sea state at each station, using the Beaufort Scale and measured by visual observation of multiple observers. These data suggest a possible inverse relationship

between plastic count/weight and the sea state. More data are needed to examine this relationship more thoroughly. However such a relationship has been tentatively identified in Kukulka et al. (2012) from MEK inhibitor review the North Atlantic. In general, changes between some pairs of samples can be explained by changes in sea surface conditions. For example, lower (compared to adjacent samples) counts in samples 27 and 28 coincide in time with the brief and sharp increase in winds. Analogously, high counts and weights in samples 22 and 23 were

obtained during a short period of weaker wind. The statistical model used herein (Maximenko et al., 2012), based on observed trajectories of drifting buoys, was successfully used to find an accumulation zone of plastic pollution in the SPSG. While this model identifies regions of maximum aggregation of the floating debris, it fails to predict the relative abundance of plastic between different gyres. For example, it predicts the maximum density in the South Pacific to be as much as ten times higher than the maximum density in the North Atlantic. The actual selleck screening library particle abundance in the central region of the North Atlantic, reported between 29 and 31°N, was 20,328 ± 2324 pieces km−2 (Law et al., 2010), i.e. the abundance was 1.3 times higher in the South Pacific (26,898 ± 60,818 pieces km−2 in this study). This is explained by the setup of the model experiment.

The relative maximum in the model South Pacific is dictated by the larger amount of tracer “injected” there in the model due to the larger size of this subtropical gyre. In reality, northern gyres appear to contain more plastics, corresponding to higher rates of production, consumption and loss of plastic to the marine environment in the northern hemisphere (Lebreton et al., 2012). Law et al. (2010) observed no significant increase in plastic marine pollution in a 22-year survey second of the North Atlantic subtropical gyre, while during a similar time frame Goldstein et al. (2012) observed a dramatic increase of microplastics in the NPSG. Overall, the densities of microplastics found in the SPSG are comparable with those observed in other areas of the world (Hidalgo-Ruz et al., 2012). They are, however, lower than those reported for the North Pacific Subtropical Gyre (NPSG). Using a similar approach as Moore et al. (2001), herein we found a mean of ∼25,000 microplastics km−2 compared to ∼330,000 microplastics km−2 in the NPSG. The maximum density in the NPSG was ∼970,000 microplastics km−2 (Moore et al.

05 with stratification according to previous TNF antagonist status, concomitant corticosteroid use, and concomitant immunosuppressive use. The Cochran–Mantel–Haenszel chi-square P value, risk difference (primary test), and associated 2-tailed 95% confidence intervals (CIs) were determined, as were the relative risk and its 2-tailed 95% CI. Secondary analyses were performed sequentially, with a P value of .05 or less required to proceed to

testing of each subsequent outcome. Of the 6 secondary analyses, 4 (ie, 2 pairs of outcomes, each pair evaluating 1 end point for the 2 populations) involved simultaneous testing for the TNF antagonist–failure and overall populations ( Supplementary Figure 1). The Hochberg method was applied to each secondary outcome pair to maintain the overall type 1 error rate at a P value of .05 or less. A logistic regression model, including baseline

CDAI score, stratification factors, and geographic CDK activation region, was conducted as a sensitivity analysis using the chi-square test at a statistical significance level of 0.05; the chi-square P value and odds ratio, with associated 95% CIs, were determined. Analysis of covariance models of change from baseline to week t for the continuous efficacy outcome variables in the vedolizumab and placebo groups 5-FU in vitro was performed. For the prespecified exploratory analyses of TNF antagonist–naive patients and for those based on concomitant corticosteroid or immunosuppressive use, P values were determined and 95% CIs were calculated using the exact method (for categoric data with numerators ≤5) or the normal approximation. Power estimates for the primary and secondary outcomes were 91% and 81%–93%, respectively, on the basis of total sample sizes of 296 for the TNF antagonist–failure population and 396 for the overall population. A total of 660 patients were screened (Figure 2), of whom 244 were excluded because of not meeting enrollment criteria (n = 209), withdrawal of consent (n = 11), having an SAE (n = 5), having

a protocol violation (n = 1), or other/unknown reasons (n = 18). Parvulin Of 416 randomized patients, 315 (76%) had previous failure of (ie, inadequate response to, loss of response to, or intolerance of) 1 or more TNF antagonists, and 101 patients (24%) were TNF-antagonist naive. Demographic characteristics (Table 1) generally were similar between treatment groups in the TNF antagonist–failure population. Corticosteroids were the most common concomitant medications used at any time during the study (54% of patients), followed by immunosuppressives (34%) and mesalamine (31%). Previous immunosuppressive exposure was reported by 89% of patients. In the TNF antagonist–failure population, 2 or more TNF antagonists had failed in 66% of patients (44% of whom had a primary nonresponse), whereas 3 TNF antagonists had failed in 11% of patients.

Jedoch muss ihr Einfluss auch unter Bedingungen des Eisenüberschusses berücksichtigt werden. Überschüssiges intrazelluläres Eisen wird in Ferritin eingebaut, ein oligomeres Protein aus 24 identischen (oder ähnlichen) Staurosporine supplier Untereinheiten mit einem Molekulargewicht von ungefähr 500 kDa. Ferritin kann bis zu 4500 Eisenionen pro

Molekül in einer nicht toxischen, aber dennoch bioverfügbaren Form binden. Die Funktion des Ferritins ist es, den Umfang des potenziell schädlichen „intrazellulären labilen Eisenpools” zu beschränken und gleichzeitig Eisen in einer Form zu speichern, das bei Knappheit mobilisiert werden kann, wodurch das Risiko für einen Eisenmangel verringert wird [25]. Die Bindung wird über die Messung der intrazellulären Konzentration des labilen Eisens durch das Iron regulatory protein/Iron responsive element-(IRP/IRE)-System an den Bedarf gekoppelt. Dieses System schränkt die Expression von Ferritin ein, wenn die intrazelluläre Konzentration HDAC inhibitor an labilem Eisen niedrig ist, und steigert die Ferritinexpression bei hoher Konzentration. Es muss angenommen werden, dass etwas labiles Eisen im Zytoplasma vorhanden ist, da solch ein System sonst nicht

funktionieren könnte; in der Tat wurde es in Zellkultur auch nachgewiesen [26] and [27]. Die Konzentration labiler Eisenionen in Zellen und im Interstitialraum kann Fenton-Reaktionen auslösen und die Balance hin zu vermehrtem oxidativem Stress verschieben [2] (zum Mechanismus siehe Abb. 2). In Kulturzellen nimmt der labile Eisenpool parallel zum oxidativen Stress zu. Beides ist mit entsprechenden intrazellulären Sonden untersucht worden, wobei die Expression der schweren Ferritin-Kette durch genetischen Eingriff inhibiert war [26]. Umgekehrt vermindert eine Überexpression der schweren Ferritinkette den labilen Eisenpool und gleichzeitig den oxidativen Stress [27]. Eiseninduzierter oxidativer Stress war in Tierexperimenten an der Pathophysiologie entzündlicher Darmerkrankungen [28] und der rheumatoiden Arthritis [29] beteiligt. Protein tyrosine phosphatase Außerdem

induziert systemische Entzündung katabole Reaktionen im Intermediärstoffwechsel [30], von denen angenommen wird, dass sie Wachstumsverzögerungen verursachen [31]; verzögertes Wachstum zeigte sich bei ausreichend mit Eisen versorgten Kindern, die mit Eisen supplementiert wurden [14] and [32]. Außerdem wird das labile Eisen als Erklärung für den verschärfenden Effekt der Eisensupplementation auf den klinischen Verlauf der Malaria herangezogen; die Plasmodien können nämlich während ihrer intraerythrozytären Phase kein Eisen aus Häm mobilisieren [33] and [34]. Die Virulenz anderer intrazellulärer Pathogene, wie z. B. Mycobakterium tuberculosis und Mycobakterium leprae, hängt ebenfalls von der Verfügbarkeit intrazellulären Eisens ab [35]. Das IRP/IRE-System steigert die zelluläre Eisenaufnahme, indem es die Expression des Transferrin-Rezeptors (TfR) mittels posttranskriptionaler Mechanismen erhöht [36].

pin mutants have greatly impaired fertility MK-2206 price and striking sporophytic defects that are similar to published defects arising from treatment with auxin transport inhibitors. Our results show that PIN proteins are conserved auxin transport facilitators. To clarify the roles of auxin in moss gametophore development, we grew colonies on medium supplemented with auxins that have different biochemical properties: indoleacetic acid (IAA), naphthylacetic acid (NAA), and 2,4-dichlorophenoxyacetic acid (2,4-D). Although weak effects were seen with the native auxin IAA (Figure S1 available online), a spectrum of phenotypes of lesser-to-greater severity was observed in treatments

with NAA and 2,4-D and was classified into five phenotypic classes, classes I–V (Figures 1A and S1). An increased frequency of more-severe phenotypes correlated with increasing auxin concentrations (Figure S1C). When grown on lower auxin concentrations (e.g., 100 nM NAA, 1 μM 2,4-D), class I and class II shoots were prevalent. Class I shoots appeared similar to controls, but the zone of rhizoid emergence was displaced apically, as in previous reports [47, 48 and 49]. Class II shoots (seen in 2,4-D treatments) were elongated and had more leaves than controls (Figures 1A, 1C, S1A,

and S1D). Class III shoots were stunted, producing fewer leaves than Selleck NVP-AUY922 untreated controls (Figures 1A, 1B, and S1D), and leaves were narrow with fewer, longer cells than untreated controls (Figures 1C, S1B, and S1D). In class IV shoots, leaf outgrowth was suppressed, and gametophores comprised a raspberry-like dome of cells above a zone of rhizoid emergence (Figure 1A). Confocal microscopy revealed

a spiral of successively larger leaf progenitor cells emanating from the apical cell, thus demonstrating its continued activity (Figure 1B). The strongest effect of auxin was revealed in class V shoots, which lost apical cell function. Shoots terminated with irregularly shaped cells, or rhizoids, consistent with previous reports [47 and 49] (Figure 1B). These data suggest that accumulation of auxin in shoots triggers diverse developmental effects at different threshold G protein-coupled receptor kinase levels. Notably, auxin accumulation causes defects in meristem function, leaf initiation, and oriented leaf growth. By analogy to flowering plants, we hypothesized that gametophore development is normally driven by changes in the auxin distribution within tissues, which was disrupted by adding exogenous auxin. We reasoned that such changes might occur by a conserved transport-dependent mechanism. To test this hypothesis, we analyzed the effect on gametophore development of the compounds 1-N-naphthylphthalamic acid (NPA) and naringenen (Nar), which are potent PATIs in angiosperms.