For patients with diffuse colonic disease but without rectal involvement, it may also be possible to consider a total abdominal colectomy with ileal rectal anastomosis. Advantages of this operation generally include preserved rectal and sexual function. The operation itself is shorter and less extensive. However, this operation does not treat dysplasia or inflammatory disease within the rectum. This

area will require continued surveillance, and in patients with both Crohn’s disease and UC the rates of recurrence selleck chemical of inflammatory disease in the rectum are as high as 60%.28 This operation is contraindicated in patients with rectal or anal lesions, and considered as very high risk for patients with multifocal dysplasia. Other contraindications include patients with baseline fecal incontinence

or severe rectal inflammation. check details For patients who are not fit for anastomosis, or reconnection, a total abdominal colectomy with Hartmann procedure may be performed. This operation leaves the remnant rectum in place during the operation, and an end ileostomy is performed. Advantages of this surgery include decreased time and morbidity by leaving the rectum in situ. However, risks include inflammation and risk of dysplasia within the rectum, and continued surveillance is necessary. In isolated inflammatory and dysplastic disease, or in cases of a sporadic adenoma, the most appropriate operation may be a segmental colectomy. Benefits of this operation include shorter operative times, maintenance of key portions of the colon, including possibly the ileocecal valve which may functionally decrease risks of diarrhea, and the greater part of the colon for fluid absorption. This option is restricted to patients with isolated dysplasia and those with relatively normal mucosa in terms

of inflammation; surgical anastomosis necessitates functional mucosa for creation of a colon anastomosis. Patients who undergo this option must be committed to continued colonoscopic surveillance to evaluate for metachronous lesions and the risk of continued progression of inflammatory disease. Data demonstrate that up to 40% of patients with Crohn’s disease click here will require additional colectomy at 10 years for recurrence of inflammation after segmental colectomy.29 and 30 All resections, whether segmental or complete proctocolectomies, should follow the principles of surgical oncology. A full lymphadenectomy and vessel resection with high ligation should be completed. Current data recommend resection of a minimum of 12 lymph nodes for segmental colectomy to ensure appropriate staging of tumors.31 In addition, good data also exist to affirm that the use of laparoscopic or minimally invasive surgery is beneficial for patients.32 All of the aforementioned procedures can be performed laparoscopically in experienced hands.

It has been shown that the pro-inflammatory cytokine tumour necrosis factor-α (TNFα) induces rapid phosphorylation of IκBα and its ubiquitin-induced degradation. This event is necessary for NFκB/p65 to be released from the complex with IκBα and for its relocation to the nucleus where it exerts transactivation functions by binding co-activator proteins [reviewed in [39]]. As

incubation with PCP resulted in decreased phosphorylation of IKKβ-mediated phosphorylation of NFκB/p65 at the activating S536, we addressed the question whether PCP affected the TNFα-mediated translocation of NFκB/p65 into the nucleus. As shown in Fig. Selleck LDK378 7, treatment of both cell lines with TNFα led to the accumulation of NFκB/p65 in the nucleus with respect to DMSO- and PCP-treated cells, respectively, where NFκB/p65 appeared

to localize in the cytoplasm. However, incubation of cells with PCP suppressed TNFα-induced migration of NFκB/p65 into the nucleus as indicated by the persistent signal in the cytoplasm. The study presented here, indicates that PCP is the active component of C11 exerting cytotoxic effects in human pancreatic cancer BTK inhibitor concentration cell lines. Our previous studies showed that simultaneous silencing of the CK2 catalytic subunits by RNA interference enhances the sensitivity of these cell lines towards chemotherapeutic agents currently used in the clinics for the cure of advanced pancreatic cancer [[5], for a review see [40]]. We show here that PCP inhibits recombinant human CK2 in an ATP-competitive manner as well as the endogenously expressed enzyme as revealed by the decreased phosphorylation of the molecular chaperone Cdc37 at S13, a known cellular substrate target of CK2 [Fig. 6., [38]]. Evidence indicates that CK2 supports survival and confers resistance to chemotherapeutic treatment of cancer cells [reviewed in [2] and [6]]. Hence, PCP-mediated induction of cell death in human pancreatic cancer cells reported here, may be due, at least partially, to the inhibition of endogenous

CK2. However, as protein kinase CK2 expression levels have been shown to be elevated in cancer and highly proliferating cells, we cannot exclude that other types of cancer cells would respond to PCP treatment in a similar fashion. Cediranib (AZD2171) The poor prognosis of pancreatic cancer is in part attributed to the presence of a subgroup of cancer stem cells which account for tumour recurrence due to their self-renewal, metastatic potential and resistance to cytotoxic drug treatment [19] and [20]. Interestingly, we show here that incubation of a sub-population of Panc-1 cells enriched in cancer stem cells with PCP induces a level of cytotoxicity comparable to the one observed in the cancer stem cells-depleted population suggesting that PCP treatment lowers the intrinsic resistance of cancer stem cells to cell death induction (Fig. 2).

Most probably the N-terminal region of the metalloproteinase domain of native moojenin under non-reducing conditions cannot be determined by Edman degradation because it is blocked by the presence of pyroglutamic acid, as is usually observed for other SVMPs of the PIII subclass (Muniz et al., Selleck AZD5363 2008). The proteolytic fragment was present in a low proportion compared to the unprocessed moojenin; however, it could be detected by sequenator analysis since this procedure presents higher sensitivity than SDS-PAGE. The proteolytic activity of the moojenin was assayed on bovine fibrinogen. Moojenin degraded fibrinogen, as evidenced by the appearance of new protein bands at the bottom of the gel. Apparently,

moojenin completely degraded the Aα-chain and Bβ-chain of fibrinogen, in a time-dependent manner ( Fig. 3A). The Aα-chain was totally degraded even at the shortest time tested (15 min), while the Bβ-chain was degraded at the longest time (90 min). The γ-chain appeared

unaffected throughout the incubation period examined. The optimal temperature range for the degradation of the fibrinogen chains was determined to be 30–40 °C. Activity was completely lost at temperatures ≥50 °C ( Fig. 3C). Apoptosis Compound Library The digestion pattern of the moojenin was similar to other purified metalloproteinases from bothropic venom, for example, BleucMP from Bothrops leucurus ( Gomes et al., 2011), BlaH1 from Bothrops lanceolatus (Fer-de-lance) ( Stroka et al., 2005) and BmooMPα-I from B. moojeni ( Bernardes et al., 2008). All these enzymes are classified as α-fibrinogenases. They degrade the Aα-chain of fibrinogen first, followed by the Bβ-chain, and show no effect on the γ-chain. SVMPs are usually more active at pHs ranging from neutral to basic (Manning, 1995;

Xu et al., 2004). Interestingly, for the first time, we demonstrated the action of a proteinase at acidic pH. Moojenin degraded fibrinogen chains at pH 4, but not at pHs ranging from neutral to basic (Fig. 3B). Chelating agents such as EDTA, 1,10 phenanthroline and β-mercaptoethanol inhibited the fibrinogenolytic MycoClean Mycoplasma Removal Kit activity of the enzyme. In contrast, benzamidine, leupeptin and PMSF did not affect this activity (Fig. 3D). These results suggest that moojenin belongs to the class of metalloproteinases and disulfide bonds are important for the maintenance of its structure. Numerous snake venom proteinases have been isolated and characterized (Serrano and Maroun, 2005). These enzymes affect, for example, fibrinogenolysis, platelet aggregation, the complement system, blood pressure and blood coagulation (Markland, 1998; Zhang et al., 1998; Castro et al., 2004; Kini, 2005; Serrano and Maroun, 2005). Interestingly, moojenin presented a coagulant activity. These results are in accordance with the finding of Serrano and colleagues (Serrano et al., 1993b). These authors purified a metalloproteinase, denominated MPB, with a residual coagulant activity.

Figure 6, Figure 7, Figure 8 and Figure 9 present the results for each group of pigments. The problem of the adaptation of phytoplankton cells to light conditions in the Baltic Sea is more complex than in Case this website 1 (ocean) waters. The relative errors of the approximated concentrations of different pigment groups are larger than for ocean waters. The only exception is chlorophyll c, for which the logarithmic statistical

error was about 8.8% lower (σ– = 34.6% for Baltic waters and 38.2% for ocean waters). Analysis of the approximated concentrations of other PSP groups, i.e. chlorophyll b and PSC, as a function of spectral fitting showed that the relative estimation errors were more than twice as large for the Baltic data than for BAY 73-4506 the ocean data. This may have been due to the different distributions of the relative spectral irradiances at different depths in Case 1 and Case 2 waters. In the deeper regions of oligotrophic waters (such as ocean waters), light comes mainly from the blue-green part of the spectrum, whereas in eutrophic waters (such as Baltic waters), there is much less of this light. The chromatic acclimation factor gives a relatively good estimate of the concentrations of the major groups of PSP in

ocean waters. But the large estimation errors in Baltic waters may be due to the phycobilin concentration modifying the light field spectrum in the Baltic, which is not taken into account in the analysis. Analysis of the errors resulting from the approximations of the PPC content, depending on the energy characteristics of the underwater irradiance in the short-range part of PAR ( eq. (7)), showed that the relative errors are ID-8 1.3 times

higher for Baltic waters than for ocean waters. The logarithmic statistical errors are σ– = 38.4% for Baltic waters and 32.0% for ocean waters. In summary, the problem of the adaptation and acclimation of phytoplankton cells to the irradiance conditions in Case 2 waters, such as those of the Baltic Sea, appears to be more complex than in Case 1 (ocean) waters. Only in the case of certain pigments does the verification of the approximations of their concentrations or the environmentally dependent concentrations of pigment groups give lower estimation errors than those resulting from the approximations found for oceanic waters. This is the situation we are faced with when estimating the total content of chlorophylls c and PPC with respect to the optical depth and the total content of chlorophylls c with respect to chromatic adaptation factors. The spectral fitting function, i.e. the chromatic adaptation factor, approximates the content of the major groups of photosynthetic pigments in ocean waters fairly well.

e. mostly tuna data), and do not mark them with the ‘F’ symbol for estimated figures. Secondly, starting with the publication of

1996 data [6], the Yearbook included only the production from capture fisheries with the exclusion of aquaculture production and its title was changed accordingly from “Catches and landings” to “Capture production”. The 1984–1997 aquaculture data had been published yearly as “FAO Fisheries Circular No. 815” but in 2000 the first FAO Aquaculture production yearbook was issued [7]. Backward revision of the two data series was completed in 2003, when fully separated capture and aquaculture datasets for the 1950–2001 period were made available through the BMN673 FISHSTAT+ software. Finally, in 2008 the three Fishery Statistics Yearbooks on “Capture production”, “Aquaculture production”, and “Fishery Commodities” have no longer been published in hard copy but only on

a CD-ROM enclosed in a booklet [8] including summary tables for all databases. Since the following edition [9] were also added overviews, charts and a section on “Food Balance Sheets”. To coordinate fishery PI3K inhibitor statistical programs of regional and inter-governmental organizations, in 1960 the FAO Conference established the “Continuing Working Party on Fishery Statistics in the North Atlantic Area” (CWP). In 1995, the CWP changed its title to “Coordinating Working Party on Fishery Statistics” due to its new global coverage. The CWP has played a key role in establishing and harmonizing concepts, techniques, classifications and standards for the collection, processing and dissemination of fishery statistics [10]. Nowadays, 19 regional and global

organizations1 participate in the mechanism meeting approximately every two years. Catch data and other fishery statistics are generally submitted to FAO by national correspondents in the appropriate ministry or institution. At about May every year, FAO sends to correspondents paper and electronic versions of standard questionnaires and encourages reporting through them. However, to facilitate data submission, any format in which the national statistics are stored is accepted by FAO. The deadline to return data to FAO is the 31st August. As soon after this date, FAO starts to send out reminders and contact those countries which have not yet submitted their data. The FAO capture database Phosphoprotein phosphatase is usually closed at about the end of February and at the beginning of March the updated database is made available on the web.2 Statistics made available by national authorities are complemented or replaced if better data of other origins are available. The CWP at its 18th Session [11] recommended members to regard as the most reliable data those held by the Regional Fishery Body (RFB) with assessment responsibility for a given stock, which are supposed to be the ‘best scientific estimate’. Following this recommendation, FAO often replaces the data received from national offices with those validated by RFBs, e.g.

15 and 0.2 (5 ml/gradient) of NaCl in buffer A (1 g of brain; 1.5 g DEAE cellulose; 5 ml NaCl/gradient). Their proteins were determined by Bradford (1976) method and calpain activity was found in the fraction of 0.2 M NaCl for brain. Calpain activity was analyzed as described

by Buroker-Kilgore and Wang (1993) as modified (Emerick et al., 2010 and Emerick et al., 2012a). To assess neurotoxicity development, a five-point scale was used as described elsewhere (DeOliveira et al., 2002): 0 indicates a normal hen; 1 is slightly abnormal gait; 2 mild ataxia; 3 severe ataxia accompanied by frequent collapse; and 4 complete incapacitation, that is, inability to move and permanent lateral recumbent. The hens were observed on days Selleck Talazoparib 8, 10, 12, 14, 16,

18 and 21 after OP intoxication, but values presented in Table 2 are scores of the twenty-first day of observation. Differences in biochemical BMS-354825 analyses were examined for statistical significance by one way ANOVA (Analysis Of Variance) followed by Tukey’s test for multiple comparisons. These tests were performed in Microsoft Office Excel® 2007 for Windows. Differences in neurotoxicity scores (non-parametric data) were tested for statistical significance with the Kruskal–Wallis test, followed by the Wilcoxon Mann–Whitney test for multiple comparisons. The non-parametric tests were carried out in the BioEstat® 5.0 program (Mamirauá, Brazil). The definition of significance was p < 0.05 for all statistical analyses. All biochemical data are presented as the averages of three samples done in triplicate (3 hens). All biochemical next data are expressed as means ± the standard deviation (SD). All clinical data are presented as the sum of score of three hens 21 days after OP treatment. Measurements were made of the activities of NTE, AChE and calpain

in samples collected from hens fasted for 12 h before euthanasia. Control values were used for the data presented in Fig. 1, Fig. 2 and Fig. 3 and are 27.2 ± 4.9 μmol/min/g of protein, 904 ± 98 μmol/min/g of protein and 16.1 ± 3.4 units of absorbance/min/g of protein for NTE, AChE and calpain in hen brain, respectively. Fig. 1 shows that only the group of hens given TOCP 500 mg/kg had NTE inhibition above 80% when compared to the control group 24 h after dosing. Among the isoforms of methamidophos, only the (+)-methamidophos (50 mg/kg) was capable of inhibiting NTE activity (approximately 60%) at that time. This inhibition was statistically significant different compared to the control group and the group that received TOCP (500 mg/kg). However, no significant differences among the groups were noted 21 days after administration of the toxicants. Fig. 2 shows that all isoforms of methamidophos at a dose of 50 mg/kg caused inhibition of AChE of approximately 80% compared to control group. The group which received TOCP 500 mg/kg inhibited the AChE activity approximately 20% compared to control.

Further, because paraspinal and PS muscles have different nerve supplies (dorsal vs. ventral rami of lumbar nerves, respectively) buy Bortezomib and MFI is increased bilaterally, denervation is not considered a plausible explanation in the current study. Finally, the positive correlation between fatty infiltration and episode frequency (mean: 4.4, min: 2, max: 9 per year; R2 = 0.450), may suggest a role for nociception in fatty infiltration.

This assumption is consistent with previous observations of generalized inhibition of MF, ES and PS recruitment with experimentally-induced pain ( Dickx et al., 2008; D’Hooge et al., submitted for publication). Further research is required to determine if peripheral nociception is involved in fatty infiltration via a reflex-mediated decrease in neural drive. Previously, Hultman

et al. (1993) found no difference in paraspinal muscle density on CT during remission of intermittent LBP. Results of fatty infiltration in the presence of LBP are less consistent than CSA measures. Some authors demonstrate increased fatty infiltration (Parkkola et al., 1993; Hultman et al., 1993; Mengiardi, 2006; Kjaer et al., 2007), whereas others show no difference to healthy controls (McLoughlin et al., 1994; Danneels et al., 2000; Kjaer et al., 2007). The discrepancy in results may be due to methodological check details differences such as the ROI in which fatty infiltration is determined (total vs. lean muscle, isolated MF vs. paraspinals grouped) or measuring technique (qualitative vs. quantitative, CT vs. MRI). The current study measured fatty infiltration

in two complementary modes yielding divergent results: lean fatty infiltration was increased, without macroscopic alterations. Similarly, Mengiardi (2006) revealed increased metabolic fat content with proton MR spectrocoscopy, which was not detectable with a semi-quantitative visual grading system using conventional MRI. Using a multifaceted approach to investigate lumbar muscle structure, the current study showed that fatty infiltration in lean muscle tissue was increased, without alterations in muscle size or macroscopic fat deposition during Arachidonate 15-lipoxygenase remission of LBP. This emphasizes the importance of differentiating muscle quantity (CSA) and quality (composition). In this respect, Elliott et al. reported enlarged cervical muscle CSAs and fatty infiltration in relation to whiplash-associated disorders, acknowledging that caution must be exercised during interpretation of CSA measurements in the presence of intramuscular fat (Elliott et al., 2008a, 2010). Similarly, lean fatty infiltration may be masking a reduction in muscle size in our results. It is assumed that fatty infiltration may negatively affect muscle contractility when muscle fibers are replaced with non-contractile tissue. Consequently, the deteriorated muscle composition may contribute to LBP recurrence.

The univariate Searchlight revealed that individual variability was larger in the situational non-translation (SnT) language-switching condition than in the focused simultaneous translation (FST) language-switching condition. In the SnT session, the informative voxels were spread in the bilateral occipital, temporal lobe, and some discrete

regions. In contrast, the results of the FST were concentrated along the routes connecting regions around the left fusiform, left and right lingual and left supramarginal gyri. In FST, all of the participants showed a similar trend, with a coherent and intense band of sensitivity. This result suggests that in the relatively difficult FST language-switching task, the participants Pirfenidone clinical trial needed

more attentive control, and so the activations of the brain were more intense and regulated. Note that the lingual region is believed to play a role in visual search and attentional control during language switching (Wang et al., 2007). An interesting finding is that the Searchlight did not detect any important voxels in the frontal lobe. buy 17-AAG In contrast, GLM detected a significant activation in the frontal region for the k2k-vs-c2c and k2c-vs-c2k conditions. Because Korean uses an alphabetic writing system, the activations in the left middle frontal gyrus (Broca’s area), left precentral and left caudate might be related to alphabetic reading. In contrast, it is possible that the clusters of informative voxels and significant activations found in the occipital lobe by both the Searchlight and GLM (c2k-vs-k2c) methods during the presentation of the Chinese stimuli were due to the logographic aspect of the Chinese character stimuli (Liu and Perfetti, 2003, Siok et al., 2004, Tan et al.,

2001 and Wang et al., 2007). Furthermore, Crinion et al. (2006) found that the left caudate played a role in monitoring and controlling bilinguals’ use of languages, which is also endorsed by our GLM result from k2k-vs-c2c. Left temporal activation may be related to general language processing, while activation in the right Dimethyl sulfoxide temporal gyrus (k2c-vs-c2k) may be related to attentional demand required for language processing (Sabri et al., 2008). Literature investigating language switching has also implicated the left fusiform. Notably, an investigation by Abutalebi et al. (2007) that applied auditory stimuli to detect language switching demonstrated that the left BA37 (-38,-25,-18) was important for controlling lexical-semantic processing. Other studies illustrated that activity in the fusiform gyrus might be indicative of some other cognitive processes (Guo et al., 2011, Hernandez, 2009, Hernandez and Meschyan, 2006 and Price et al., 1999; Moritz-Gassera & Duffau, 2009). Investigations using invasive techniques (Duffau et al., 2014, Kho et al.

3). We then investigated whether http://www.selleckchem.com/products/VX-765.html the bacterial infection interfered with ovary activation in the beebread-fed queenless bees. Infection indeed impaired ovary activation, as was shown by a significantly lower number of bees with activated ovaries compared to the non-infected bees on this same diet (Fig.

3, insert). To investigate whether the effects of nutrition and infection extended to other reproduction-related genes (in addition to storage protein and receptor genes), we analyzed the vasa transcripts levels in the bees fed on different diets and challenged with S. marcescens. Significantly higher vasa transcripts levels were observed in the bees fed beebread than in those fed the other diets ( Fig. 4). Like observed for the vg, vgR, and hex 70a genes, bacterial infection impaired the increase in vasa transcript levels in the beebread-fed AZD2014 cell line bees ( Fig. 4). In the present study, we explored the costs of bacterial infection on gene transcription, protein storage and ovary activation in honey bee workers in relation to the type of the supplied diet. In a previous study (Lourenço et al., 2009), we used injection rather than oral administration to bacterially infect bees and then analyzed vg and hex 70a expression at 12 h post-infection. The transcript and protein-level responses to bacterial injection

were not distinguishable from those caused by water injection (injury). In the present work, the injury effect was circumvented by orally administering the bacteria via the diet. In addition, we extended the duration of the experiments (to 6 and 9 days) and considered additional parameters, i.e., nutrition and ovary status (activated or non-activated). Three other genes (vgR, apoLpR and vasa) were also investigated in the current study. Notably, the cost of infection on transcription and protein levels was mostly evident in the beebread-fed bees. In these bees, the transcription of vg, vgR, hex 70a and vasa, and the levels of Vg and Hex 70a proteins, were clearly impaired by the infection. These results indicate

that the physiological cost of infection is better evidenced under certain dietary conditions. Furthermore, the dynamic process of Vg storage (in hemolymph) and mobilization (to the fat body) may have been disrupted since the expression of vgR was inhibited in beebread-fed however bees as a consequence of the infection. Royal jelly, like beebread, is a rich source of proteins for bees. It might be thought that the proportion of royal jelly in the diet was insufficient to allow increased levels of vg, vgR, hex 70a and vasa transcripts, and the Vg and Hex 70a proteins. Alternatively, the diet could have provided an excess of royal jelly and caused adverse effects on transcription. It is known that high levels of dietary protein consumption negatively correlate with survival in young worker honey bees ( Pirk et al., 2010).

1). Ears at 10, 15, 20, 22, 25 and 30 days after pollination (DAP) were collected from plants grown under standard greenhouse

conditions. Kernels were dissected from the ears, immediately frozen in liquid nitrogen, and stored at − 80 °C until RNA extraction. Isolated total RNA was size-fractionated on a 15% Tris–borate–EDTA (TBE) urea polyacrylamide gel to enrich molecules of 15–30 nt. The small RNA was ligated with adapters (5′-GTCTCTAGCCTGCAGGATCGATG-3′) AG-014699 supplier and (5′-AAAGATCCTGCAGGTGCGTCA-3′), and (5′-GTCTCTAGCCTGCAGGATCGATG-3′) and (5′-AAAGATCCTGCAGGTGCGTCA-3′) using T4 RNA ligase and size-fractionated on a 15% TBE urea polyacrylamide gel. The resultant RNA was reversely transcribed to cDNA with a small RNA RT-primer (5′-CAAGCAGAAGACGGCATACGA-3′), and the cDNA was then directly subcloned into vector pMD18-T (TaKaRa). These tandem cDNA fragments were transformed into Escherichia Selleck Selumetinib coli strain DH5 by electroporation. Colony PCR was performed using 5′ and 3′ primers, and clones with lengths of 60–80 bp were used for sequencing according to the manufacturer’s protocols (Colony PCR Made Easy,

http://www.lucigen.com/colonyPCR). Small RNAs (200 nt) were isolated with the mirVana PARIS Kit (Ambion) according to the manufacturer’s instructions. For reverse transcription (RT), 1 μg of small RNA was treated with the miScript Reverse Transcription Kit (Qiagen) at 37 °C for 60 min and a final incubation at 95 °C for 5 min. Real-time PCR of miRNAs was carried out Interleukin-2 receptor using the miScript SYBR Green PCR kit (Qiagen) in an Applied Biosystems 7500 real-time PCR machine (ABI). PCR was conducted at 95 °C for 15 min, followed by 40 cycles of incubation at 94 °C for 15 s, 55 °C for 30 s, and then 70 °C for 30 s. Each PCR was repeated at least three times. All samples were normalized to 5S rRNA expression and fold change expression was calculated according to the 2− ΔΔCt method as described previously [39]. High-quality small RNA reads larger than 18 nt were extracted from the raw reads and mapped to maize genome sequences (http://www.maizesequence.org) using SOAPaligner/soap2

(http://soap.genomics.org.cn/soapaligner.html) [40]. Matched sequences were then queried against non-coding RNAs from the Rfam database (http://www.sanger.ac.uk/Software/Rfam) and the ncRNA database (http://www.ncrna.org/frnadb/blast/fRNAdb). Most non-miRNAs, non-siRNAs and mRNA degradation fragments were removed by a BLASTn search of the NCBI GenBank database (http://www.ncbi.nlm.nih.gov/blast/Blast.cgi) [41]. Any small RNAs with exact matches to these sequences were excluded from further analysis. miRNAs were predicted with Mireap (https://sourceforge.net/projects/mireap/). Secondary structures of the predicted miRNAs were confirmed using the RNAfold online tool (http://rna.tbi.univie.ac.at/cgi-bin/RNAfold.cgi).