Rterial pressures for control and experimental groups. No statistically significant differences were observed among mean arterial pressures during 5day treatment period. doi:10.1371/journal.pone.0046568.tTNF with or without LOS did not have any effect on blood pressure parameters (Table 2).EchocardiographyWhen compared to control animals, TNF Felypressin custom synthesis treated rats had progressive increases in LV end-diastolic dimension (LVD), LV end-systolic dimension (LVS) and TEI index and decreases in fractional shortening (FS ) measurements (Table 3). MNS Additionally, rats treated with TNF+LOS exhibited significant decreases in LVD, LVS and TIE index and increases in FS when compared to rats given TNF only.EPR Measurements Enzymatic and Respiratory Activities of Mitochondrial Complexes I IIMeasurements of mitochondrial complex I, II and III enzymatic activities were performed as previously described [12]. Briefly, aliquots of mitochondria were mixed with oxygenated KHB (20 mm Hg O2) containing 1 mM EGTA. Then, the oxygen spin label NOX-13.1-OS (5 mM), CMH (200 mM), and one of the following substrates were added to the mitochondrial suspension: 20 mM glutamate (complex I), 5 mM succinate (complex II), or 5 mM pyruvate (complex III). After adequate mixing, the samples were taken into capillary tube and mitochondrial complex I, II and III were measured using EPR. Activity of each mitochondrial complex was quantified by EPR under the same settings described previously [12,22]. Total ROS, O2N2 and 25837696 OONO2 production rates in LV tissue, as determined by EPR, were all significantly higher in LV tissues of TNF treated rats than in control and TNF+LOS groups (Figure 1a?c ). Increases in ROS generation induced by TNF or ANGII were significantly inhibited by LOS. These results further support the ability of LOS to reduce oxidative stress in LV tissue. Table 3. Echocardiographic data from experimental groups.Parameter IVSD (mm) IVSS mm) LVD (mm) LVS (mm) PWD(mm) PWS (mm) FSControl 1.3760.001 2.3160.08 7.2760.06 5.1960.10 1.2960.02 2.1860.026 36.8660.72 332.2563.2 0.2760.TNF 1.6860.06 2.4860.07 7.9760.06* 5.7460.16* 1.4260.04 2.2460.09 26.8560.45* 329.4064.1 0.3860.04*TNF +LOS 1.3360.06 2.3760.1 7.3460.1 5.2760.1 1.3460.1 2.2760.04 35.8860.7 333.7564.9 0.2960.003Measurement of ATP and ADP/ATP RatioATP production rates and ADP/ATP ratios were quantified in isolated mitochondria using a commercially available kit (BioVision, Mountain View, CA) as described previously [12,22].Statistical AnalysesAll data are expressed as mean 6 SEM. Statistical analyses were performed using GraphPad Prism version 5.00 for Windows. Data were analyzed by ANOVA, followed by Bonferroni’s multiple comparison tests. In all cases, p,0.05 was considered statistically significant.HR TEIResults Blood PressureBlood pressure measurements were obtained for all experimental animals during the 5 day treatment period. Administration ofEchocardiographic analysis revealed that both left ventricular diastolic (LVD) and systolic (LVS) dimensions were significantly greater in TNF treated animals. TNF treatment also decreased fractional shortening (FS) and increased Tei index when compared to controls. Co treatment of TNF treated rats with losartan prevented these changes. IVSD, intraventricular septal thickness at end diastole; IVSS, intraventricular septal thickness at end systole; PWD, posterior wall thickness at end diastole; PWS, posterior wall thickness at end systole; HR, heart rate. Values are expressed.Rterial pressures for control and experimental groups. No statistically significant differences were observed among mean arterial pressures during 5day treatment period. doi:10.1371/journal.pone.0046568.tTNF with or without LOS did not have any effect on blood pressure parameters (Table 2).EchocardiographyWhen compared to control animals, TNF treated rats had progressive increases in LV end-diastolic dimension (LVD), LV end-systolic dimension (LVS) and TEI index and decreases in fractional shortening (FS ) measurements (Table 3). Additionally, rats treated with TNF+LOS exhibited significant decreases in LVD, LVS and TIE index and increases in FS when compared to rats given TNF only.EPR Measurements Enzymatic and Respiratory Activities of Mitochondrial Complexes I IIMeasurements of mitochondrial complex I, II and III enzymatic activities were performed as previously described [12]. Briefly, aliquots of mitochondria were mixed with oxygenated KHB (20 mm Hg O2) containing 1 mM EGTA. Then, the oxygen spin label NOX-13.1-OS (5 mM), CMH (200 mM), and one of the following substrates were added to the mitochondrial suspension: 20 mM glutamate (complex I), 5 mM succinate (complex II), or 5 mM pyruvate (complex III). After adequate mixing, the samples were taken into capillary tube and mitochondrial complex I, II and III were measured using EPR. Activity of each mitochondrial complex was quantified by EPR under the same settings described previously [12,22]. Total ROS, O2N2 and 25837696 OONO2 production rates in LV tissue, as determined by EPR, were all significantly higher in LV tissues of TNF treated rats than in control and TNF+LOS groups (Figure 1a?c ). Increases in ROS generation induced by TNF or ANGII were significantly inhibited by LOS. These results further support the ability of LOS to reduce oxidative stress in LV tissue. Table 3. Echocardiographic data from experimental groups.Parameter IVSD (mm) IVSS mm) LVD (mm) LVS (mm) PWD(mm) PWS (mm) FSControl 1.3760.001 2.3160.08 7.2760.06 5.1960.10 1.2960.02 2.1860.026 36.8660.72 332.2563.2 0.2760.TNF 1.6860.06 2.4860.07 7.9760.06* 5.7460.16* 1.4260.04 2.2460.09 26.8560.45* 329.4064.1 0.3860.04*TNF +LOS 1.3360.06 2.3760.1 7.3460.1 5.2760.1 1.3460.1 2.2760.04 35.8860.7 333.7564.9 0.2960.003Measurement of ATP and ADP/ATP RatioATP production rates and ADP/ATP ratios were quantified in isolated mitochondria using a commercially available kit (BioVision, Mountain View, CA) as described previously [12,22].Statistical AnalysesAll data are expressed as mean 6 SEM. Statistical analyses were performed using GraphPad Prism version 5.00 for Windows. Data were analyzed by ANOVA, followed by Bonferroni’s multiple comparison tests. In all cases, p,0.05 was considered statistically significant.HR TEIResults Blood PressureBlood pressure measurements were obtained for all experimental animals during the 5 day treatment period. Administration ofEchocardiographic analysis revealed that both left ventricular diastolic (LVD) and systolic (LVS) dimensions were significantly greater in TNF treated animals. TNF treatment also decreased fractional shortening (FS) and increased Tei index when compared to controls. Co treatment of TNF treated rats with losartan prevented these changes. IVSD, intraventricular septal thickness at end diastole; IVSS, intraventricular septal thickness at end systole; PWD, posterior wall thickness at end diastole; PWS, posterior wall thickness at end systole; HR, heart rate. Values are expressed.

ncer, including EOC, has begun to be more fully appreciated. The most studied epigenetic alteration is DNA methylation, the addition of a methyl moiety to the cytosine-5 position within the context of a CpG dinucleotide, mediated by DNA methyltransferases. DNA methylation patterns are reset early in the embryogenesis and reestablished early during development. After that, they are thought to be relatively stable. In cancer, the physiological regulation of DNA methylation is disrupted leading to drastic changes of the distribution pattern of 5-methylcytosine. The heavy methylation found in the bulk of chromatin is reduced, while the normally unmethylated CpG islands located in the promoter and first exon of genes become hypermethylated. Promoter hypermethylation often leads to inactivation of different tumor-suppressing genes and is associated with many important pathways involved in cancer, such as DNA repair, cell cycle regulation, apoptosis, carcinogen metabolism, hormonal response, and cell adherence. Aberrant DNA methylation is also involved in the development of resistance to chemotherapy . The role of DNA hypomethylation in carcinogenesis is less studied. Recent studies have demonstrated that global decrease in the level of DNA methylation is related to hypomethylation of repeated sequences, increase in genetic instability, as well as reactivation of proto-oncogenes and pro-MEK 162 metastasis genes. Similar to other malignancies, aberrant DNA methylation, including global hypomethylation of heterochromatin and local CpG island methylation, occurs in EOC and contributes to ovarian tumorigenesis and mechanisms of chemoresistance. Applying a more global array-based technology, several studies have demonstrated that DNA methylation changes in ovarian cancer are cumulative with disease progression and CT resistance. Using a similar approach we have recently shown that DNA hypermethylation occurs in less invasive/early stages of ovarian tumorigenesis, while advanced disease was associated with DNA hypomethylation of a number of oncogenes, implicated in cancer progression, invasion/ metastasis and probably chemoresistance. The polypeptide N-acetylgalactosaminyltransferase 3 gene was among the genes identified to be notably hypomethylated in lowmalignant potential and high grade serous EOC tumors. The GALNT3 gene is a member of the GalNAc-transferases gene family; the genes of this family conduct the transfer of N-acetyl galactosamine to the hydroxyl group of a serine or threonine residue in the first step of O-linked oligosaccharide biosynthesis. So far, 20 members of the GALNAC-Ts gene family have been identified and most of them encode an active polypeptide GALNT www.impactjournals.com/oncotarget 545 functioning in the primary step of the O-glycosylation of different proteins, including mucins. Aberrant mucintype O-glycosylation represents one of the most abundant posttranslational cancer-associated changes, comprising diverse biologic and pathologic consequences influencing growth and survival of cancer cells and their ability for invasion and metastasis. The membrane-associated mucin-1, as one highly glycosylated protein, is overexpressed in more than 90% of high-grade PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19858123 EOC tumors including metastatic lesions, as MUC1 expression correlated with EOC progression. This prompted us to further investigate if GALNT3 displays elevated expression levels in serous EOC tumors with different malignant potential, and whether this gene is functionally imp

Gentamycin at 15 mg/ml.Figure 5. The blaA gene is induced by ampicillin only at high levels. Cultures of late-exponential phase cells (,0.6 of OD600) were diluted 1:100 with LB broth containing ampicillin at H (50 mg/ml), M (2.5 mg/ml), or L (0.125 mg/ml) amounts, and incubated at 30uC in a shaker at 200 rpm (A) PblaA promoter activities were determined by measuring b-galactosidase (in Miller units) using the PblaA-lacZ reporter system in the wild type. Results are averages of at least three replicates, and the error bars represent standard deviation (SD). Activity of PblaA in the presence of ampicillin at the H (50 mg/ml) level were also assayed using qRT-PCR (presented as diamonds). (B) b-lactamase activity assay. At the indicated times, samples were taken for b-lactamase activity measurements. In both panels, experiments were performed at least in triplicate and the error bars represent standard deviation (SD). doi:10.1371/journal.pone.0060460.gExpression of blaA in S. oneidensisFigure 6. Impacts of the loss of LMW PBPs on growth in the presence of ampicillin. (A) Susceptibility assay of LMW PBP mutants ndacB (PBP4), ndacA (PBP5), and npbpG (PBP7) to ampicillin. ndacAc represents the ndacA strain complemented in trans. (B) Growth of the ndacA strain in the presence of ampicillin at H (50 mg/ml), M (2.5 mg/ml) or L (0.125 mg/ml) levels. H-WT and M-WT represent growth of the wild type strain under the specified conditions. (C) Activities of PblaA-lacZ in strains devoid of one of the LMW PBPs. After growth for two hours, samples were taken for bgalactosidase measurements. Experiments were performed at least in triplicate and the error bars represent standard deviation (SD) as in (B and C). doi:10.1371/journal.pone.0060460.gConstruction and complementation of in-frame deletion mutantsIn-frame deletion mutants were constructed using the fusion PCR method was as previously described [43]. Primers used in this study are listed in Table S1. Each deletion mutation was verified by sequencing of the mutated region. For genetic complementation, either promoterless pHG101 or its derivative pHG102, which contains the S. oneidensis arcA promoter for genes not in proximity to their promoter, was used [29]. Introduction of each verified complementation vector into the corresponding mutant was achieved by mating with E. coli WM3064 containing the vector, and confirmed by plasmid extraction, restriction enzyme mapping and sequencing.were aliquotted into 24-well plates with a volume of 2 ml per well. Antibiotics and natural products were added to each well at three concentrations. The plates were kept at the room temperature for observation. The morphology of cells was examined with a Motic BA310 phase-contrast microscope. Micrographs were captured with a Moticam 2306 charged-coupled-device camera and Motic images advanced 3.2 software. All experiments were conducted at least in triplicate.Antibiotic susceptibility assayAntibiotic susceptibility of S. oneidensis was determined with both liquid and solid cultures. For antibiotics commonly used in genetic manipulation, the highest concentrations were set according to the molecular 86168-78-7 Indolactam V cost biology manual and lower concentrations were prepared by double dilution. Three ml of ISC cultures were spotted onto LB agar plates containing antibiotics of varying concentrations. The plates were incubated for up to 3 days and scored for growth each day. No growth, some growth after 3 days, and full growth after 1 day were co.Gentamycin at 15 mg/ml.Figure 5. The blaA gene is induced by ampicillin only at high levels. Cultures of late-exponential phase cells (,0.6 of OD600) were diluted 1:100 with LB broth containing ampicillin at H (50 mg/ml), M (2.5 mg/ml), or L (0.125 mg/ml) amounts, and incubated at 30uC in a shaker at 200 rpm (A) PblaA promoter activities were determined by measuring b-galactosidase (in Miller units) using the PblaA-lacZ reporter system in the wild type. Results are averages of at least three replicates, and the error bars represent standard deviation (SD). Activity of PblaA in the presence of ampicillin at the H (50 mg/ml) level were also assayed using qRT-PCR (presented as diamonds). (B) b-lactamase activity assay. At the indicated times, samples were taken for b-lactamase activity measurements. In both panels, experiments were performed at least in triplicate and the error bars represent standard deviation (SD). doi:10.1371/journal.pone.0060460.gExpression of blaA in S. oneidensisFigure 6. Impacts of the loss of LMW PBPs on growth in the presence of ampicillin. (A) Susceptibility assay of LMW PBP mutants ndacB (PBP4), ndacA (PBP5), and npbpG (PBP7) to ampicillin. ndacAc represents the ndacA strain complemented in trans. (B) Growth of the ndacA strain in the presence of ampicillin at H (50 mg/ml), M (2.5 mg/ml) or L (0.125 mg/ml) levels. H-WT and M-WT represent growth of the wild type strain under the specified conditions. (C) Activities of PblaA-lacZ in strains devoid of one of the LMW PBPs. After growth for two hours, samples were taken for bgalactosidase measurements. Experiments were performed at least in triplicate and the error bars represent standard deviation (SD) as in (B and C). doi:10.1371/journal.pone.0060460.gConstruction and complementation of in-frame deletion mutantsIn-frame deletion mutants were constructed using the fusion PCR method was as previously described [43]. Primers used in this study are listed in Table S1. Each deletion mutation was verified by sequencing of the mutated region. For genetic complementation, either promoterless pHG101 or its derivative pHG102, which contains the S. oneidensis arcA promoter for genes not in proximity to their promoter, was used [29]. Introduction of each verified complementation vector into the corresponding mutant was achieved by mating with E. coli WM3064 containing the vector, and confirmed by plasmid extraction, restriction enzyme mapping and sequencing.were aliquotted into 24-well plates with a volume of 2 ml per well. Antibiotics and natural products were added to each well at three concentrations. The plates were kept at the room temperature for observation. The morphology of cells was examined with a Motic BA310 phase-contrast microscope. Micrographs were captured with a Moticam 2306 charged-coupled-device camera and Motic images advanced 3.2 software. All experiments were conducted at least in triplicate.Antibiotic susceptibility assayAntibiotic susceptibility of S. oneidensis was determined with both liquid and solid cultures. For antibiotics commonly used in genetic manipulation, the highest concentrations were set according to the molecular biology manual and lower concentrations were prepared by double dilution. Three ml of ISC cultures were spotted onto LB agar plates containing antibiotics of varying concentrations. The plates were incubated for up to 3 days and scored for growth each day. No growth, some growth after 3 days, and full growth after 1 day were co.

Rimp, qRT-PCR was conducted using isoform-specific primers and probes. The isoform Ago1C was shown to be ubiquitously expressed in all organs or tissues examined (Fig. 4A). However, the mRNA levels of both Ago1A and Ago1B were significantly up-regulated in lymphoid organ (Fig. 4A), suggesting that the two isoforms were involved in shrimp immunity. Considering that Ago BI 78D3 web proteins represent key components in antiviral RNAi immunity in many species [12,13,14,15], the effects of WSSV infection on the expression of Ago1 isoforms was investigated. Because of the up-regulation of Ago1A and Ago1B 25033180 isoforms in shrimp lymphoid organs, the lymphoid organ was selected to investigate the expression profiles of Ago1 isoforms in response to WSSV challenge. It was showed that the expression of Ago1A and Ago1B was significantly increased at 12 and 24 h postinoculation (p,0.05) (Fig. 4B), whereas the expression of Ago1C remained unchanged at all time points compared to the controlResults Identification of Ago1 Isoforms in ShrimpBased on PCR amplification using degenerated primers and RACE analysis, full-length cDNA of the shrimp Ago1 gene was obtained. Sequence analysis revealed that the Ago1 gene generated three transcripts: Ago1A (GenBank accession no.: GU265732), Ago1B (GenBank accession no.: JX170715) and Ago1C (GenBank accession no.: JX170716). Sequence comparisons showed that the Ago1 isoforms were different from each other with a 9-nt inserted fragment (Ago1-fragment 1) at their 59 termini and an additional 81-nt fragment of (Ago1-fragment 2) in the PIWI domain (Fig. 1). No amplification was observed when the extracted RNA was used in PCR analyses. These findingsRole of Z-360 site Argonaute-1 Isoforms in Antiviral DefenseRole of Argonaute-1 Isoforms in Antiviral DefenseFigure 6. Roles of Ago1 isoforms in the shrimp immune response against WSSV infection. To characterize the roles of Ago1 isoforms in the antiviral immunity, shrimp were injected with WSSV and isoform-specific siRNAs. Shrimp were injected simultaneously with WSSV and the low or high concentration of Ago1A-siRNA (A), Ago1B-siRNA (B), Ago1A/B-siRNA (C) or Ago1C-siRNA (D), respectively. As control, WSSV+control siRNA and WSSV only were included in the injections. At 48 h post-inoculation, the shrimp from each treatment were subjected to quantitative real-time PCR to quantify the expressions of Ago1A, Ago1B, and Ago1C (left). The solutions used for injections were shown in the box. At the same time, the WSSV loads in shrimp were monitored by quantitative real-time PCR (right). The statistically significant differences between treatments were represented with asterisk (*P,0.05). Lane headings showed the solutions used for injections. doi:10.1371/journal.pone.0050581.g(0 h post-inoculation) (Fig. 4B). Taken together, these results indicated that Ago1A and Ago1B isoforms that contained the Ago1-fragment 2 played important roles in shrimp antiviral immunity.Effects of Ago1 Isoforms on Shrimp Antiviral ImmunityTo investigate the roles of Ago1 isoforms in antiviral immunity, the expression of Ago1 isoforms were each silenced in shrimp using isoform-specific siRNAs, followed by WSSV challenge. First, to test the specificities of Ago1 isoform-specific siRNAs, FLAGtagged Ago1 isoform constructs and isoform-specific siRNAs were transfected into S2 cells. Western blot analysis showed that the expression of Ago1A, Ago1B or Ago1C isoforms was inhibited by the corresponding sequence-specific Ago1A-siRNA, Ago1BsiRNA.Rimp, qRT-PCR was conducted using isoform-specific primers and probes. The isoform Ago1C was shown to be ubiquitously expressed in all organs or tissues examined (Fig. 4A). However, the mRNA levels of both Ago1A and Ago1B were significantly up-regulated in lymphoid organ (Fig. 4A), suggesting that the two isoforms were involved in shrimp immunity. Considering that Ago proteins represent key components in antiviral RNAi immunity in many species [12,13,14,15], the effects of WSSV infection on the expression of Ago1 isoforms was investigated. Because of the up-regulation of Ago1A and Ago1B 25033180 isoforms in shrimp lymphoid organs, the lymphoid organ was selected to investigate the expression profiles of Ago1 isoforms in response to WSSV challenge. It was showed that the expression of Ago1A and Ago1B was significantly increased at 12 and 24 h postinoculation (p,0.05) (Fig. 4B), whereas the expression of Ago1C remained unchanged at all time points compared to the controlResults Identification of Ago1 Isoforms in ShrimpBased on PCR amplification using degenerated primers and RACE analysis, full-length cDNA of the shrimp Ago1 gene was obtained. Sequence analysis revealed that the Ago1 gene generated three transcripts: Ago1A (GenBank accession no.: GU265732), Ago1B (GenBank accession no.: JX170715) and Ago1C (GenBank accession no.: JX170716). Sequence comparisons showed that the Ago1 isoforms were different from each other with a 9-nt inserted fragment (Ago1-fragment 1) at their 59 termini and an additional 81-nt fragment of (Ago1-fragment 2) in the PIWI domain (Fig. 1). No amplification was observed when the extracted RNA was used in PCR analyses. These findingsRole of Argonaute-1 Isoforms in Antiviral DefenseRole of Argonaute-1 Isoforms in Antiviral DefenseFigure 6. Roles of Ago1 isoforms in the shrimp immune response against WSSV infection. To characterize the roles of Ago1 isoforms in the antiviral immunity, shrimp were injected with WSSV and isoform-specific siRNAs. Shrimp were injected simultaneously with WSSV and the low or high concentration of Ago1A-siRNA (A), Ago1B-siRNA (B), Ago1A/B-siRNA (C) or Ago1C-siRNA (D), respectively. As control, WSSV+control siRNA and WSSV only were included in the injections. At 48 h post-inoculation, the shrimp from each treatment were subjected to quantitative real-time PCR to quantify the expressions of Ago1A, Ago1B, and Ago1C (left). The solutions used for injections were shown in the box. At the same time, the WSSV loads in shrimp were monitored by quantitative real-time PCR (right). The statistically significant differences between treatments were represented with asterisk (*P,0.05). Lane headings showed the solutions used for injections. doi:10.1371/journal.pone.0050581.g(0 h post-inoculation) (Fig. 4B). Taken together, these results indicated that Ago1A and Ago1B isoforms that contained the Ago1-fragment 2 played important roles in shrimp antiviral immunity.Effects of Ago1 Isoforms on Shrimp Antiviral ImmunityTo investigate the roles of Ago1 isoforms in antiviral immunity, the expression of Ago1 isoforms were each silenced in shrimp using isoform-specific siRNAs, followed by WSSV challenge. First, to test the specificities of Ago1 isoform-specific siRNAs, FLAGtagged Ago1 isoform constructs and isoform-specific siRNAs were transfected into S2 cells. Western blot analysis showed that the expression of Ago1A, Ago1B or Ago1C isoforms was inhibited by the corresponding sequence-specific Ago1A-siRNA, Ago1BsiRNA.

The diagnostic criteria and the studied populations are very different from one study to another (Tables 5 and 6), these analyses suggest that both CMV and HSV are associated with increased Autophagy mortality rates. This study strongly suggests that CMV reactivation in critically ill patients is associated with increased mortality. With respect to HSV infections, its impact on various outcome measures seems to be less important when compared to patients infected with CMV. However, only a trial evaluating the efficacy of an anti-viral treatment in ICU patients could demonstrate that CMV and/or HSV alter outcome.Author ContributionsConceived and designed the experiments: 12926553 YC LP SB BL. Performed the experiments: YC SB JMF SH BL AR CZ MM LP. Analyzed the data: YC LP SB. Wrote the paper: YC LP SJ DR.Impact of CMV and HSV on Ventilated Patients
Insulin-like Growth Factor-1 (IGF-1) is a potent peptide factor involved in a broad range of tissue processes including cell growth and survival, proliferation, differentiation and metabolism, but the molecular basis of these diverse functions is not well understood. In the adult mammal, IGF-1 is synthesized predominately in the liver, and acts as a systemic growth factor, playing important roles in both normal and neoplastic growth [1]. IGF-1 is also produced in extrahepatic tissues where it plays a predominantly autocrine/ paracrine role in local processes. Despite a significant reduction of serum IGF-1 peptide levels in mice where the Igf-1 gene was deleted conditionally in the liver, other parameters were largely normal, indicating that locally synthesized IGF-1 can support normal postnatal growth and development [2]. The diversity of IGF-1 actions may derive from the existence of several different isoforms that differ from one another due to alternative splicing of the initial transcript [3,4]. The single copy Igf-1 gene locus encodes multiple pre-propeptide precursors in which the mature protein is flanked by variable N-terminal signal peptides and C-terminal extension (E) peptides. In the mouse, the Igf-1 gene encodes four main pre-propeptides, combining signal peptides (SP1 or SP2) with Ea or Eb extension peptides (Figure 1). As these pre-propeptides all undergo post-translational processing to generate the same mature 70 aa IGF-1 protein, the specific roles of E-peptides in IGF-1 biology remain unclear. One of the isolated E-peptides (Eb, renamed MGF) has been reported to increase the regenerative capability of skeletal muscle, play a neuroprotectiverole against ischemia, and facilitate the actions of IGF-1 to improve cardiac function and mobilize resident stem cell populations [5,6,7]. Other studies suggest that E-peptides are not required for IGF-1 secretion but increase cell entry of IGF-1 from the media [8]. Transgenic studies have shed further light on the role of Epeptides. IGF-1Ea propeptide provided as a muscle-specific Epigenetic Reader Domain transgene results in muscle hypertrophy and enhances regeneration after injury [9,10,11], reducing inflammation and fibrosis [12]. This phenotype is unaffected by the choice of N-terminal signal peptide [13] but is not recapitulated by a muscle-specific transgene encoding IGF-1 lacking an E-peptide moiety, which produces no local effects but instead significantly increases serum IGF-1 levels [14]. The dramatic phenotypes resulting from supplemental tissue-specific IGF-1Ea transgene expression in other tissues such as heart [15] and skin [16], with no increase in circulating IGF-1.The diagnostic criteria and the studied populations are very different from one study to another (Tables 5 and 6), these analyses suggest that both CMV and HSV are associated with increased mortality rates. This study strongly suggests that CMV reactivation in critically ill patients is associated with increased mortality. With respect to HSV infections, its impact on various outcome measures seems to be less important when compared to patients infected with CMV. However, only a trial evaluating the efficacy of an anti-viral treatment in ICU patients could demonstrate that CMV and/or HSV alter outcome.Author ContributionsConceived and designed the experiments: 12926553 YC LP SB BL. Performed the experiments: YC SB JMF SH BL AR CZ MM LP. Analyzed the data: YC LP SB. Wrote the paper: YC LP SJ DR.Impact of CMV and HSV on Ventilated Patients
Insulin-like Growth Factor-1 (IGF-1) is a potent peptide factor involved in a broad range of tissue processes including cell growth and survival, proliferation, differentiation and metabolism, but the molecular basis of these diverse functions is not well understood. In the adult mammal, IGF-1 is synthesized predominately in the liver, and acts as a systemic growth factor, playing important roles in both normal and neoplastic growth [1]. IGF-1 is also produced in extrahepatic tissues where it plays a predominantly autocrine/ paracrine role in local processes. Despite a significant reduction of serum IGF-1 peptide levels in mice where the Igf-1 gene was deleted conditionally in the liver, other parameters were largely normal, indicating that locally synthesized IGF-1 can support normal postnatal growth and development [2]. The diversity of IGF-1 actions may derive from the existence of several different isoforms that differ from one another due to alternative splicing of the initial transcript [3,4]. The single copy Igf-1 gene locus encodes multiple pre-propeptide precursors in which the mature protein is flanked by variable N-terminal signal peptides and C-terminal extension (E) peptides. In the mouse, the Igf-1 gene encodes four main pre-propeptides, combining signal peptides (SP1 or SP2) with Ea or Eb extension peptides (Figure 1). As these pre-propeptides all undergo post-translational processing to generate the same mature 70 aa IGF-1 protein, the specific roles of E-peptides in IGF-1 biology remain unclear. One of the isolated E-peptides (Eb, renamed MGF) has been reported to increase the regenerative capability of skeletal muscle, play a neuroprotectiverole against ischemia, and facilitate the actions of IGF-1 to improve cardiac function and mobilize resident stem cell populations [5,6,7]. Other studies suggest that E-peptides are not required for IGF-1 secretion but increase cell entry of IGF-1 from the media [8]. Transgenic studies have shed further light on the role of Epeptides. IGF-1Ea propeptide provided as a muscle-specific transgene results in muscle hypertrophy and enhances regeneration after injury [9,10,11], reducing inflammation and fibrosis [12]. This phenotype is unaffected by the choice of N-terminal signal peptide [13] but is not recapitulated by a muscle-specific transgene encoding IGF-1 lacking an E-peptide moiety, which produces no local effects but instead significantly increases serum IGF-1 levels [14]. The dramatic phenotypes resulting from supplemental tissue-specific IGF-1Ea transgene expression in other tissues such as heart [15] and skin [16], with no increase in circulating IGF-1.