7±2.6 vs 13.6±2.4mmol/L; p<0.0001), while episodes of hyperglycaemia were less (median: 3 [IQR 1–8] vs 7 [IQR 4–12]; p=0.001). Patients who experienced hypoglycaemia were also less likely to have a repeat episode with the BBB protocol (median:

http://www.selleckchem.com/products/ganetespib-sta-9090.html 1 [IQR 1–3] vs 3 [IQR 2–4.5]). The BBB protocol is easy to implement and resulted in significant improvement in BGL control compared with SSI. Copyright © 2011 John Wiley & Sons. “
“The neurological complications of diabetic ketoacidosis (DKA) include cerebral oedema or, rarely, acute cerebrovascular accident (CVA) due to ischaemic brain infarction or haemorrhage. These complications result from complex haemostatic mechanisms involving a state of systemic inflammation, coagulopathy, endothelial dysfunction and loss of blood volume induced by insulin deficiency. The development of cerebral oedema is believed to be under-reported in adult patients with DKA as compared to children. Only a limited number of case reports exist in the literature regarding the development of CVA as a complication of DKA in adults. A high index of suspicion needs to be maintained for early recognition of neurological

complications as associated signs and symptoms may only be subtle and masked by altered sensorium commonly seen in the acute phase of DKA, leading to potentially catastrophic consequences if left untreated. Here we present the case of a 22-year-old man with type 1 diabetes who developed cerebellar infarction with associated brainstem herniation as a complication of diabetic ketoacidosis and required urgent neurosurgical intervention. selleck compound Copyright © 2012 John Wiley & Sons. Practical Diabetes 2012; 29(9): 377–379

“
“This study aimed to describe a diabetes specialist nurse (DSN) telemedicine advice service in a university hospital diabetes service in terms of the payment by results (PbR) tariff costs, potential admissions avoidance and casemix. The source, purpose, duration, outcome and patient age were recorded prospectively over 12 months for every patient-initiated, diabetes-related telephone consultation. Megestrol Acetate In all, 5703 patient-initiated telephone consultations were recorded. Of these, 3459 (60.7%) involved insulin dose management for those receiving insulin therapy for longer than six months. In contrast, 530 (9.3%) consultations covered dose adjustment for individuals started on insulin therapy within the previous six months. A total of 235 (4.1%) consultations involved managing insulin, food and fluid intake during intercurrent illness (‘sick day’ advice) – 103 (1.8%) with ketonuria and 132 (2.3%) without ketonuria. Of these, only 17 required referral to their general practitioner for review for a hospital admission, representing 218 potentially avoided admissions over the study period. Individuals over 60 years of age accounted for 3610 (63.3%) consultations. The PbR tariff for each telephone consultation was £23 ($37.66; €26.10), with an estimated annual cost of £131 169 ($214 781; €148 908).

However, Che1-dependent signaling is shown to contribute indirectly to surface attachment, indicating that distinct mechanisms are likely underlying flocculation and attachment to surfaces in A. brasilense. Chemotaxis is a widespread function in motile soil bacteria as it affords cells with the ability to sense and to Epacadostat order navigate toward the most favorable niches

for growth (Wadhams & Armitage, 2004). At the molecular level, the chemotaxis pathway is the dedicated chemosensory signal transduction system that allows cells to couple detection of physicochemical changes in their surroundings to changes in the swimming pattern (i.e. chemotaxis). Chemotaxis signal transduction has been best studied in Escherichia coli and experimental evidence indicates that this prototypical enteric model is conserved and functions similarly (with some variations on the theme) in phylogenetically diverse motile bacteria. In addition to regulating chemotaxis responses in motile bacteria, chemotaxis-like signal transduction pathways were shown to regulate cellular behaviors other than flagellar rotation in several other bacterial species (Kirby, 2009), including the alphaproteobacterium Azospirillum brasilense, a soil diazotroph (Bible et al., 2008).

In only a few cases, however, have the molecular targets of these chemotaxis-like pathways been identified. Seliciclib supplier The A. brasilense Che1 chemotaxis-like pathway has been shown to have a minor,

and likely indirect, function in regulating chemotaxis behavior in this species (Hauwaerts et al., 2002; Bible et al., 2008; Edwards et al., 2011). Experimental evidence indicates that Che1 functions to modulate changes in adhesive cell surface properties which impact the propensity for cell-to-cell aggregation and flocculation (Bible et al., 2008). Deletions of cheA1 or cheY1, which each code for central proteins controlling the response output of the signal transduction pathway, yield cells that aggregate and flocculate more than the wild-type strain (Bible et al., 2008). A mutant strain deleted for all of the genes encoded within the che1 gene cluster has a phenotype similar to the strains lacking only CheA1 or CheY1, consistent with a role for Che1 selleckchem in regulating the ability of cells to flocculate. A strain carrying a mutation that disrupts the function of both CheB1 and CheR1 is severely impaired in flocculation, consistent with CheB1 and CheR1 functioning in a signaling feedback loop that controls chemosensory adaptation (Stephens et al., 2006; Bible et al., 2008). Other possible roles that Che1 may have on functions such as adhesion to surfaces or root colonization, have been previously proposed to be related to flocculation (Burdman et al., 2000a, b) but have not yet been investigated. The purpose of the present study was to determine the conditions under which A.

C. Pedersen has received research funding from Abbott, Roche, Bristol-Myers Squibb, Merck Sharp & Dohme, GlaxoSmithKline, Swedish Orphan Drugs and Boehringer Ingelheim. J. Gerstoft has received research funding from Abbott, Roche, Bristol-Myers Squibb, Merck Sharp & Dohme, Pharmasia, GlaxoSmithKline, Swedish Orphan Drugs and Boehringer Ingelheim. Line D. Rasmussen, Merete Dybdal, Gitte Kronborg, Carsten S. Larsen, Gitte Pedersen, Lars Pedersen, Janne Jensen and Henrik T Sørensen report no conflicts of interest. “
“Adherence is critical for maximizing the effectiveness of pre-exposure prophylaxis (PrEP) in preventing HIV infection. Strategies for

promoting adherence to HIV treatment, and their potential Bortezomib datasheet application to PrEP adherence, have received considerable attention. However, adherence promotion strategies for prevention medications have not been well characterized and may be more applicable to PrEP. We aimed to identify adherence support interventions that have been effective in other prevention fields and could be applied in the HIV prevention context to support pill taking among PrEP users. To identify adherence support interventions that could be evaluated and applied in the PrEP context, we conducted a systematic review across the following Selleck HSP inhibitor prevention fields: hypertension, latent tuberculosis infection, hyperlipidaemia,

oral contraceptives, osteoporosis, malaria prophylaxis, and post-exposure prophylaxis for HIV infection. We included randomized controlled trials that evaluated the efficacy of interventions to improve adherence to daily oral medications prescribed for primary prevention in healthy individuals or for secondary prevention in asymptomatic individuals. Our searches identified 585 studies, of which 48 studies met the eligibility criteria and were included in the review; nine evaluated Arachidonate 15-lipoxygenase multiple strategies, yielding 64 separately tested interventions. Interventions with the strongest evidence for improving adherence included complex, resource-intensive interventions, which combined multiple adherence support

approaches, and low-cost, low-intensity interventions that provided education or telephone calls for adherence support. Our review identified adherence interventions with strong evidence of efficacy across prevention fields and provides recommendations for evaluating these interventions in upcoming PrEP studies. “
“We investigated whether age modified associations between markers of HIV progression, CD4 T lymphocyte count and HIV RNA viral load (VL), and the following markers of metabolic function: albumin, haemoglobin, high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and total cholesterol (TC). A retrospective analysis of data from the United Kingdom Collaborative HIV Cohort was carried out. Analyses were limited to antiretroviral-naïve subjects to focus on the impact of HIV disease itself.

To support recommendations in its submission, ACP developed the following: a comprehensive review of pharmacist prescribing,[3] The ACP presented the proposed expanded scope of practice to the Minister and the Health Professions Advisory Board in November 2003[8] supported by numerous organizations including the University of Alberta Faculty of Pharmacy, the National Association of Pharmacy Regulatory Authorities (NAPRA), the Pharmaceutical Examining Board of Canada (PEBC) and the Minister of Health and Wellness Enzalutamide molecular weight at that time, Ms Iris Evans. External stakeholders

also presented information to the board at that meeting. Cabinet approved Bill 22 on 30 May 2006 and it was proclaimed in force on 1 April 2007. The following explanatory analysis will describe the development and implementation of Bill 22 to the Health Professions Act (1999). The framework adopted for this analysis is proposed by Lomas,[9] including problem definition, policy development process, and consequences of implementation. There are a number of inter-related problems driving the development of this legislation. The HWRC identified the following

problems, among others: The healthcare system is inefficient because it is not flexible with respect to scopes and roles of practice. More reflective of the time in which Bill 22 was being developed, the 2001 Premier’s Advisory Council on Health for Alberta Report (Mazankowski Report) described five areas within the current healthcare structure that the province needed to address.[10] These are described in Table 1. In a direct mailing to ACP stakeholders Compound C in vitro entitled ‘Pharmacist Prescribing: The Facts’ the perceived problems which Bill 22 would

address, from the perspective of the pharmacist profession, included: Pharmacists are drug-therapy experts who are limited by existing legislation from optimizing their contribution to the healthcare system. Stakeholders who participated directly in the process of developing, or are influenced by, these Regulations are described in Table 2. The ACP[11] proposed that pharmacists be given legislative authority Nintedanib (BIBF 1120) for three activities: 1 Initial prescribing access: Prescribing when a patient chooses the pharmacist for advice and treatment of minor injuries, chronic illnesses or conditions, to support lifestyle changes, disease prevention, or for time sensitive care. For these options presented, an analysis of the knowledge taken into account in formulating this alternative, core values underpinning the policy and the relationship to the goals of the policy is provided in Table 3.[12,13] Table 4 describes the barriers and facilitators for areas of knowledge, key values, institutional structures and external influences.[14–16] Privileges for pharmacists in Alberta granted through this policy include: 1 Initial prescribing access.

An alternative explanation is that these proteins are not Tat substrates, but are translocated through another route, such as for example the Sec pathway. The next residue (Leu18 in AmyH) is also commonly a strongly hydrophobic residue, usually Leu, Ile, or Val, but changing this residue to Ala in SufI does not lead

to a block in its translocation Cyclopamine mw (Stanley et al., 2000). In contrast, it is critical in AmyH, as the L18A mutant is not translocated at all, shown both by the starch-plate assays and Western blotting (Fig. 3). This finding is corroborated by the observation that none of the haloarchaeal proteins in our datasets contained an Ala in that position. As outlined in the introduction, the haloarchaeal Tat system differs on several aspects from those of nonhalophilic Tat systems. Therefore, we could not exclude the possibility that, for instance, proteins with RK or KR motifs would also be Tat-dependent substrates. However, we found that residues that are critical to the translocation of an E. coli Tat substrate are also critical to the export of AmyH, including both arginine residues and the first of the pair of hydrophobic residues that follow the arginines. In addition, the second hydrophobic residue in the Tat motif is also essential for AmyH secretion, while

this residue seems to be of less importance in the E. coli Tat substrate SufI. The sequence logos indicate that this residue can also be another strongly hydrophobic amino acid such as Val or Ile, but further mutational

analysis has to be performed to confirm this. It is Methisazone interesting to note Wortmannin concentration that the importance of this residue was already indicated by our bioinformatics analysis. The consensus motif for haloarchaeal Tat substrates can be denoted as (S/T)RRx(F/L)L, even though the first residue (Ser or Thr) does not appear to be essential for translocation. This information is useful in the prediction of Tat substrates encoded by genes found in haloarchaeal genomes. We do need to note, though, that our conclusions are based on the analysis of only one haloarchaeal Tat substrate, and it is clear that the characterization of other signal peptides is needed to understand the requirements for Tat-dependent export fully. D.K. was sponsored by a studentship from the Biotechnology and Biological Sciences Research Council, and A.B. was supported by a University Research Fellowship from the Royal Society. Table S1. Uniprot accession numbers and their Tat motifs. Please note: Wiley-Blackwell is not responsible for &!QJ;the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. “
“Salmonella enterica serovar Enteritidis is a major cause of human gastrointestinal tract disease, infection being due in large part to the consumption of contaminated eggs. Recent genome sequencing of S.

The prevalence of tubal ligation was 27% in the study participants. Little is known about the influence of reproductive, gynecological and hormonal

factors on survival of ovarian cancer and very few studies have investigated selleck chemicals the influence of tubal ligation on ovarian cancer survival. The results from our study confirm a finding in a UK study that reported a past history of surgical sterilization to be an adverse independent prognostic indicator in women presenting with stage III epithelial ovarian cancer.10 However, another study reported that previous tubal sterilization was associated with improved survival and a decrease the cancer death risk in Danish women with Stage III ovarian carcinomas, although the association was not statistically significant.11 Two other studies, conducted in Australia and the UK respectively, reported no association of ovarian cancer survival with tubal ligation or hysterectomy.12,13 Tubal ligation has consistently been reported to predict a reduced risk of ovarian cancer incidence in epidemiological studies and is recognized as an established protective factor,2–6 which is in contrast to the observation in our study

that previous tubal ligation was an independently adverse prognostic factor Apitolisib for survival from the same cancer. Serous carcinoma is the most common epithelial ovarian malignancy.17 Most cases in the subtype present at an advanced stage and the overall prognosis is poor.21–23 The proportions of serous carcinoma accounted for 57% and 34% of the participants with and without a tubal sterilization prior to diagnosis, respectively. A higher proportion of the serous carcinoma subtype in the patients who previously had

a tubal sterilization much may partially explain its adverse influence on survival of the cancer, because that subtype of histopathology is associated with poor prognosis.21–23 A recent review and meta-analysis reported that a higher risk reduction was found for endometrioid invasive cancers in comparison with the other types. A less apparent reduction was found for serous-invasive cancers, whereas the results did not reach statistical significance for mucinous-invasive cancers.24 The hypothesis that chronic inflammation in the fallopian tube resulting from a tubal ligation may explain its adverse influence on ovarian cancer survival was proposed in other studies. One study reported that chronic inflammation in the fallopian tube was a possible risk factor for mutagenesis leading to serous carcinoma.25 Another study found that in situ epithelial lesions of the fallopian tube show gene copy abnormalities consistent with these being early lesions of serous carcinoma.26 Further studies that examine the relationship are warranted to support the hypothesis. Several issues should be taken into consideration when interpreting our results.

5-L culture was washed twice with 1 M NaCl and 10 mM EDTA, pH 7.0, and twice with double-distilled water. The pellet was dissolved in sterile water and sonicated for 5 min with 3-s pulses at 30% amplitude in a Branson digital sonifier (model 250, Branson Ultrasonics Corporation, CT).

The sonicated suspension was centrifuged at 15 000 g for 30 min. The supernatant this website was discarded and the pellet was dissolved in 50 mM NaOH. This suspension was incubated on ice for 3 h with gentle shaking. The suspension was centrifuged at 15 000 g for 20 min at 4 °C. The supernatants containing the solubilized binary toxins were dialyzed overnight against buffer A (25 mM Tris-HCl, 10 mM NaCl, 2 mM DTT, pH 9.0). The dialyzed suspension was centrifuged at 15 000 g for 20 min at 4 °C and the supernatant was loaded on a Q-sepharose column Vincristine (Bio-Rad laboratories, Hercules, CA). The bound protein was eluted with a linear gradient of 10–1000 mM NaCl over a six-column volume. The binary proteins coeluted at around 300 mM NaCl concentration. The eluted protein fractions were analyzed on 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The pure protein fractions were pooled and dialyzed extensively against buffer A. After dialysis, the pooled fractions were concentrated to ∼2 mg mL−1 and loaded on to a Superdex™ 200 10/300 GL column (GE Healthcare Bio-Sciences, Uppsala, Sweden) for further purification. The purified fractions were further resolved on 12% SDS-PAGE. The

purified protein was dialyzed against 25 mM Tris-HCl, pH 8.0, 10 mM NaCl buffer, the protein was estimated using modified Lowry’s protocol and then tested ADP ribosylation factor for toxicity against third-instar larvae of C. quinquefasciatus. Different concentrations of purified binary proteins, along with control and buffer control, were tested in 10 mL tap water containing

10 third-instar C. quinquefasciatus larvae in each beaker (10 mL), with three replications for each concentration, and experiments were repeated three times. The total larval mortality was scored after 48 h of treatment. Mortality data were analyzed using probit analysis and the LC50 values were calculated at a 95% confidence limit (spss 12.0 for Windows). TVC of indigenous isolates and standard 1593 and 2362 grown in NB medium were in the range of 3.8–13 × 108 spores mL−1 (Table 1). Among these isolates, a significantly higher TVC (F=710.99; d.f.=4; P<0.05) was obtained with ISPC-8 (1.3 × 109 spores mL−1). The results of the insecticidal activity of different B. sphaericus strains revealed varying virulence patterns against third-instar larvae C. quinquefasciatus (Table 1). The range for LC50 and LC90 values observed for indigenous isolates was 0.68–6.44 × 103 spores mL−1 and 6.85–37.40 × 103 spores mL−1, respectively, whereas the respective LC50 values for standard strains 1593 and 2362 were 1.85 and 1.22 × 103 spores mL−1 and the LC90 values were 15.39 and 20.58 × 103 spores mL−1. This observation indicates that ISPC-8 (LC50 0.

, 2004), as well as improving tactile acuity of the index finger when applied on the approximate area of its cortical representation (Tegenthoff et al., 2005). Intermittent high-frequency stimulation (iHFS), a form of repetitive peripheral tactile stimulation of the index finger, is similarly effective in improving tactile http://www.selleckchem.com/products/SP600125.html acuity (Ragert et al., 2008) and, as we show here, also increases cortical excitability. Ragert et al. (2003) demonstrated that rTMS and peripheral tactile stimulation can interact when applied simultaneously,

with one potentiating the other’s effect on tactile acuity, although their results suggested a potential ceiling limit to the combined effect, or a possible homeostatic mechanism controlling the possible range of plastic alterations. In this study, we aimed to investigate the extent to which these two interventions (rTMS and tactile iHFS) would interact when applied consecutively. Additionally, we sought to determine if two kinds of parameters, behavioural and neurophysiological, are affected by the interaction in similar ways. We

tested three groups, each with 15 subjects, who were all right-handed (20 females, aged 20–28 years; mean age, 24 years). Subjects gave their written informed consent prior to participating. The study protocol was approved by the local ethics committee of the Ruhr-University Bochum, and the project protocol was performed in accordance with the Declaration of Helsinki. To study changes in cortical excitability, we applied a paired-pulse protocol consisting

of paired electrical median nerve stimulation, selleck compound with an interstimulus interval (ISI) of 30 ms. Stimulation of the median nerve was selected in order to establish a link between the SEP recordings and the cortical representation of the right index finger selected for the two stimulation protocols (rTMS and iHFS). Nerve stimulation was performed using a block electrode placed on the wrist (pulse duration, 0.2 ms; repetition rate of the paired stimuli, 2 Hz; ISI between paired stimuli, 30 ms). The median nerve stimulation intensity was set at the motor threshold, defined as the intensity at which a visible contraction of the thenar muscles was detected, and was kept constant for each subject throughout the experiment. Subjects were asked to report a prickling Org 27569 phenomenon in the thumb, index and middle fingers of the stimulated hand in order to verify correct positioning of the stimulating block electrode. During median nerve stimulation and SEP recordings, subjects were seated in a comfortable chair, and were instructed to relax and to stay awake, with their eyes closed. SEPs were recorded and stored for offline analysis using a Schwarzer 8 apparatus (bandpass filter 2–2000 Hz). Paired-pulse SEP recordings were made using a two-electrode array. One electrode was located over the SI, 2 cm posterior to the C3 position (C3′), according to the International 10–20 system.

41 ± 5.61 and 16.77 ± 19.52, respectively. Caries activity and gingivitis were correlated with the presence of mature dental biofilm. Prevalence of soft tissue lesions, dental caries and gingivitis in HIV-infected children was high and correlated to lack of satisfactory oral hygiene habits, suggesting the need of therapeutic programmes that allow these

children to recover their oral health. “
“International Journal of Paediatric Dentistry 2012; 22: 265–270 Background.  A device based on infrared laser fluorescence (IRLF) has become available as an adjunct for the diagnosis of dental caries. Aims.  The objective of this study was to clarify the differences of IRLF readings in the mesial, central and distal occlusal pits of first permanent molars. Design.  Sixty-four children (average age 8.0 years) selleck kinase inhibitor were examined using IRLF. The mesial, central and distal pits of clinically

healthy first permanent molars were measured. The instrument provides measurements in arbitrary units on an open-ended interval scale. Results.  Mean (± SE) IRLF values in the mesial pits were 4.9 ± 0.4 (upper) and 6.5 ± 0.4 (lower) and were significantly lower than those in the central (8.8 ± 0.6 and 11.5 ± 0.9) and distal (9.6 ± 0.7 and 10.4 ± 0.8) pits in the maxilla and mandible. There was no significant difference between the right (7.3 ± 0.5, 9.4 ± 0.6) and left (8.2 ± 0.5, 9.5 ± 0.6) dental arches. IRLF measurements in the mesial pits of human first permanent sound molars were lower than the central and distal pits in children whose second molars had not erupted. IMP dehydrogenase Conclusions. 

The click here inherently higher IRLF values of some sites should not be misinterpreted and trigger early invasive treatment. “
“Child abuse and neglect (CAN) is a widespread social phenomenon encompassing all forms of maltreatment with serious lifelong consequences. Dentists and dental team members are in the unique position to identify the symptoms of CAN often visible in craniofacial region. To evaluate Croatian dentists’ level of knowledge, experience, and attitude towards CAN issue. Investigation was conducted in five major Croatian cities (Zagreb, Varaždin, Osijek, Rijeka, and Split). A previously used questionnaire regarding knowledge and experience in child protection was adopted to Croatian terminology and distributed to 544 dentists. A total of 510 dentists who returned a questionnaire with valid data 26.27% reported to have had suspicion of CAN during professional career and 5.1% reported their suspicion within the last 6 months, mostly to social services and police. Fear of violence towards the child and uncertainty about observations were the most frequently reported barriers towards referring and only 11.4% knew the procedure. About 80% of respondents want further training in identifying and reporting of physical abuse. Study showed a lack of knowledge and uncertainty in recognizing and reporting CAN cases in Croatian dentists.

, 2008). find more In an attempt to identify the target proteins affected by virB, we compared

protein differences between a virB mutant and its parental strain using comparative proteomic analysis (Wang et al., 2009). Interestingly, several intracellular survival-related proteins, including VjbR, DnaK, HtrA, Omp25 and GntR, were downregulated in the virB mutant. Of these proteins affected by virB, products of the two major outer membrane proteins (OMPs), Omp25 and Omp31, were expressed at decreased levels, implying that T4SS might affect the membrane properties of Brucella. OMPs are essential for maintaining the integrity and selective permeability of membranes (Moriyon & Lopez-Goni, 1998). In addition, OMPs are often regulated by environmental signals and play important roles in bacterial pathogenesis by enhancing the adaptability to various environments (Lin et al., LGK-974 2002; Caro-Hernandez et al., 2007). Virulence regulation systems, exemplified by VjbR and BvrR/BvrS, regulate the expression of membrane proteins. The mutants showed an altered

expression of OMPs. Because of the limited separation resolution of two-dimensional polyacrylamide gel electrophoresis (2-DE), only a small part of the proteins could be isolated and identified. Therefore, it is possible that far more OMPs are differentially expressed in the virB mutant and that OM-related phenotypes are altered. To further test the effect of T4SS on the OM, in the present study, OMPs of a wild-type and a virB Etofibrate mutant strain were isolated and compared. The membrane integrity was tested by comparing the sensitivity of these proteins to polymyxin B and several stresses. Notably, a large number of OMPs were differentially expressed. More protein products of Omp25 and Omp31 were shown to be altered, revealing a complicated post-translational modification of the two proteins. In vitro sensitivity assays showed that the resistance of the virB mutant to different stress

environments was reduced. These data indicated that a drastic modification in the OM of the virB mutant occurred and that T4SS plays important roles in membrane integrity. A virB inactivation mutant BMΔvirB (BM with a promoter of the virB operon deleted) and complementary strains BM-IVGT (BMΔvirB containing complementary plasmid pBBR1-IVGT) were constructed previously (Wang et al., 2009). Brucella was cultured in tryptic soy broth (TSB) or tryptic soy agar (TSA). When necessary, antibiotics were added to a final concentration of 100 μg mL−1 ampicillin and 25 μg mL−1 gentamicin. The Brucella OM fractions were isolated as described previously (Ying et al., 2005). 2-DE and matrix-assisted laser desorption/ionization time-of-flight(MALDI-TOF) MS were performed essentially as described previously (Wang et al., 2009). Total RNA was isolated with Trizol agent (Invitrogen, Carlsbad, CA) as recommended by the manufacturer.