6). Chronic infection with HBV has been recognized to exacerbate liver fibrosis in patients.7-10 Mouse models for liver fibrosis have been successfully established in normal mice31-33 but there was no animal model to mimic liver fibrosis occurring in long-term HBV-infected patients. In this work, to our knowledge, we are the first to observe spontaneously

occurring or CCl4-induced liver fibrosis in HBV-tg mice, and thus explored the possible immunologic RAD001 mechanisms. The oversensitive liver fibrosis induced by CCl4 in HBV-tg mice may help us to investigate the precise mechanisms of liver fibrosis during chronic HBV infection. A question always exists as to the relationship between liver injury and fibrosis. Although liver injury is not the only pathway involved in liver fibrosis, for example, HSCs might be directly activated without an intermediate step of aggressive liver injury through PDGF overexpression,34 in general, the severity and persistence of liver injury determines the outcome of liver fibrosis. In our study, liver fibrosis followed chronic liver injury. In Fig. 1 we show the spontaneously developed liver fibrosis (increased

transcription of α-SMA, transcription of col1a1, MMP2, and TIMP1) accompanied by liver injury (elevated serum ALT) in 6-month-old find more HBV-tg mice. In Figs. 2-5 it is shown that in chronic CCl4-induced liver fibrosis, HBV-tg mice also had more liver fibrosis associated with more injury. Generally, it was realized that cytotoxic T lymphocytes (CTLs) contribute to initiate hepatocyte injury.35, 36 However,

the effector 上海皓元 mechanisms are not only by CTLs but also by other immune cells, among which the roles of innate immune cells in CTL-related or -unrelated inflammatory-mediated fibrosis is unclear and needs study. In CTL-related injury, the CTL-derived cytokines might activate other innate immune cells (such as NKT cells) to produce more inflammatory cytokines, which indirectly lead to hepatocyte injury in addition to CTL direct-killing hepatocytes.37 On the other hand, in CTL-unrelated injury, previously we and others found that innate cells (NK, NKT cells) mediated liver injury in HBV transgenic mice (a mouse model without CTL function).38, 39 In our experiments with respect to HBV-related liver fibrosis, we found NKT cells are pivotal to activate HSCs (Figs. 7, 8). The accumulating data indicate that NKT cells could be activated through TCR recognition (e.g., Vα14/Vβ8 in mice) with antigen-CD1d complex (usually glycolipid) or other killer cell receptors such as NKG2D of NKT cells with their ligands (Rae-1, Mult-1).

6). Chronic infection with HBV has been recognized to exacerbate liver fibrosis in patients.7-10 Mouse models for liver fibrosis have been successfully established in normal mice31-33 but there was no animal model to mimic liver fibrosis occurring in long-term HBV-infected patients. In this work, to our knowledge, we are the first to observe spontaneously

occurring or CCl4-induced liver fibrosis in HBV-tg mice, and thus explored the possible immunologic Wnt inhibitor mechanisms. The oversensitive liver fibrosis induced by CCl4 in HBV-tg mice may help us to investigate the precise mechanisms of liver fibrosis during chronic HBV infection. A question always exists as to the relationship between liver injury and fibrosis. Although liver injury is not the only pathway involved in liver fibrosis, for example, HSCs might be directly activated without an intermediate step of aggressive liver injury through PDGF overexpression,34 in general, the severity and persistence of liver injury determines the outcome of liver fibrosis. In our study, liver fibrosis followed chronic liver injury. In Fig. 1 we show the spontaneously developed liver fibrosis (increased

transcription of α-SMA, transcription of col1a1, MMP2, and TIMP1) accompanied by liver injury (elevated serum ALT) in 6-month-old Selleckchem Midostaurin HBV-tg mice. In Figs. 2-5 it is shown that in chronic CCl4-induced liver fibrosis, HBV-tg mice also had more liver fibrosis associated with more injury. Generally, it was realized that cytotoxic T lymphocytes (CTLs) contribute to initiate hepatocyte injury.35, 36 However,

the effector 上海皓元医药股份有限公司 mechanisms are not only by CTLs but also by other immune cells, among which the roles of innate immune cells in CTL-related or -unrelated inflammatory-mediated fibrosis is unclear and needs study. In CTL-related injury, the CTL-derived cytokines might activate other innate immune cells (such as NKT cells) to produce more inflammatory cytokines, which indirectly lead to hepatocyte injury in addition to CTL direct-killing hepatocytes.37 On the other hand, in CTL-unrelated injury, previously we and others found that innate cells (NK, NKT cells) mediated liver injury in HBV transgenic mice (a mouse model without CTL function).38, 39 In our experiments with respect to HBV-related liver fibrosis, we found NKT cells are pivotal to activate HSCs (Figs. 7, 8). The accumulating data indicate that NKT cells could be activated through TCR recognition (e.g., Vα14/Vβ8 in mice) with antigen-CD1d complex (usually glycolipid) or other killer cell receptors such as NKG2D of NKT cells with their ligands (Rae-1, Mult-1).

h-α-SMA was more strongly expressed than mouse α-SMA, as measured by real-time polymerase chain reaction (PCR), and in drug-treated animals the human isoform of α-SMA but not the murine isoform was down-regulated, suggesting that injected PTFs were still present and functionally active at the end of the experiment, and also that the presence of host/resident myofibroblasts did not significantly affect results (Fig. 6D). In conclusion, we demonstrated that LPA secreted by HCC cells recruits

and activates PTFs, orchestrating their differentiation Roxadustat to a CAF-like myofibroblastic phenotype which, in turn, accelerates HCC progression. Finally, we aimed to validate these findings in HCC patients. We therefore analyzed LPA serum levels in 60 patients with HCC (30 patients with and 30 without metastases), and in 50 patients with liver cirrhosis. We found that LPA serum levels were higher in HCC compared with liver cirrhosis patients (P < 0.05). Among HCC patients, Selleck STA-9090 LPA serum levels were significantly (P < 0.05) higher in those with metastasis compared with those

without (Fig. 6A). Patients with higher (P < 0.001) serum levels of LPA also have larger HCC tumors (>4 cm) and shorter survival compared with those with lower LPA serum concentrations (Fig. 6B,C). To validate our LPA data in HCC patients, publicly accessible microarray data were analyzed for a correlation between ATX and clinical outcome in HCC patients. ATX expression was significantly increased in HCC patients with more advanced disease, in particular in those with metastatic invasion (P < 0.001) (Fig. 6D),13 and was more strongly expressed in tumoral compared with paired nontumoral tissues (Fig. 6E,F) . In addition, we compared the expression of ATX and LPA receptors in epithelial and stromal components of the same HCC tissues microdissected

using the laser capture microscope technique 上海皓元 (Fig. 7A,B). We found similar expression levels of ATX in both the epithelial and the stromal component of HCC. However, LPA receptors were essentially expressed in the stromal rather than the epithelial component, indicating the stroma as the main target of the LPA paracrine loop (Fig. 7C). Finally, the ACTA2 gene was significantly expressed in tumoral compared with paired nontumoral tissues (Fig. 7D). This is consistent with publicly accessible microarray data published by Wang (Fig. 7E). Taken together, these data show that the stromal component represents the main target of LPA in patients with HCC, and that α-SMA–positive cells are predominant within the tumor stroma, as shown by the increased expression of the ACTA2 gene.

h-α-SMA was more strongly expressed than mouse α-SMA, as measured by real-time polymerase chain reaction (PCR), and in drug-treated animals the human isoform of α-SMA but not the murine isoform was down-regulated, suggesting that injected PTFs were still present and functionally active at the end of the experiment, and also that the presence of host/resident myofibroblasts did not significantly affect results (Fig. 6D). In conclusion, we demonstrated that LPA secreted by HCC cells recruits

and activates PTFs, orchestrating their differentiation MLN8237 in vitro to a CAF-like myofibroblastic phenotype which, in turn, accelerates HCC progression. Finally, we aimed to validate these findings in HCC patients. We therefore analyzed LPA serum levels in 60 patients with HCC (30 patients with and 30 without metastases), and in 50 patients with liver cirrhosis. We found that LPA serum levels were higher in HCC compared with liver cirrhosis patients (P < 0.05). Among HCC patients, check details LPA serum levels were significantly (P < 0.05) higher in those with metastasis compared with those

without (Fig. 6A). Patients with higher (P < 0.001) serum levels of LPA also have larger HCC tumors (>4 cm) and shorter survival compared with those with lower LPA serum concentrations (Fig. 6B,C). To validate our LPA data in HCC patients, publicly accessible microarray data were analyzed for a correlation between ATX and clinical outcome in HCC patients. ATX expression was significantly increased in HCC patients with more advanced disease, in particular in those with metastatic invasion (P < 0.001) (Fig. 6D),13 and was more strongly expressed in tumoral compared with paired nontumoral tissues (Fig. 6E,F) . In addition, we compared the expression of ATX and LPA receptors in epithelial and stromal components of the same HCC tissues microdissected

using the laser capture microscope technique 上海皓元医药股份有限公司 (Fig. 7A,B). We found similar expression levels of ATX in both the epithelial and the stromal component of HCC. However, LPA receptors were essentially expressed in the stromal rather than the epithelial component, indicating the stroma as the main target of the LPA paracrine loop (Fig. 7C). Finally, the ACTA2 gene was significantly expressed in tumoral compared with paired nontumoral tissues (Fig. 7D). This is consistent with publicly accessible microarray data published by Wang (Fig. 7E). Taken together, these data show that the stromal component represents the main target of LPA in patients with HCC, and that α-SMA–positive cells are predominant within the tumor stroma, as shown by the increased expression of the ACTA2 gene.

We did not consider multivariate analysis because of the wide heterogeneity and lack of complete data for identification of possible variables that could explain heterogeneity. A chi-squared for interaction was used to examine whether the 1-year survival varied significantly between subgroups.

Begg’s funnel plots were generated, and Egger’s regression asymmetry test was used to examine potential publication bias related to the 1-year survival rates. For all analyses, P < 0.05 was considered statistically significant. All analyses were completed with SAS version 8.1 (SAS Institute, selleck products Cary, NC) software. This study was not supported by any pharmaceutical company or grants; the cost was borne by the authors’ institutions. After review of the titles and abstracts, 30 RCTs8–37 fulfilled the inclusion criteria and were selected for review. Twenty studies9, 12–21, 23, 25, 26, 28, 31, 33–35, 37 were North American and European, and 108, 10, 11, 22, 24, 27, 29, 30, 32, 36 were Asian-Pacific. Of the 30 RCTs, 148–21 were

published before 2000, and the other 1622–37 since 2000. The distribution of the main characteristics of patients in the control arm of the 30 RCTs8–37 considered in the current analysis is reported in Table 1. Angiogenesis inhibitor Characteristics of arms (treatment and control) of RCTs included in the meta-analysis are detailed in Supporting Table 2. In 15 RCTs, there was an inactive placebo arm,12, 15–17, 19, 24, 25, 29, 30, 32–37

whereas in the others, untreated patients received no treatment or supportive care only.8–11, 13, 14, 18, 20–23, 26–28, 31 A total of 4335 patients were included in these 30 studies, 1927 of whom were in the control group. The size of the control groups in each study ranged from 1112 to 30335 patients. The percentage of men ranged from 6526 to 100.11 Mean patient age was 62.3, ranging from 4911 to 69.34, 37 The proportion MCE公司 of patients with cirrhosis ranged from 6334 to 100%.12, 19, 20, 23 Data on the cause of liver disease were missing in many trials. HCV status was not reported in 11 trials,8–12, 17, 22, 24, 27, 30, 37 and anti-HCV, when reported, was positive in 436 to 94%13 of the patients. HBV status was not reported in six trials,9, 11, 12, 22, 30, 37 and hepatitis B surface antigen, when reported, was positive in 013, 23 to 94.4%.10 The proportion of patients with alcohol-related liver disease was not reported in 13 RCTs,8, 10–12, 18, 22, 24, 26, 27, 30, 32, 34, 36 and ranged from 2.525 to 78%31 in studies reporting alcohol consumption. Among the studies providing data on the distribution of the ECOG Performance Status (ECOG PS),13, 16, 17, 20, 27, 28, 30, 31, 32, 35–37 the frequency of an ECOG PS = 0 went from 032 to 77%.

This suggests that bowheads have a sense of smell, and we speculate that they may use this to find aggregations of krill on which they feed. “
“Aerial photographs were analyzed to investigate the feeding habits of the Bering-Chukchi-Beaufort (BCB) population of bowhead whales (Balaena mysticetus), particularly

epibenthic feeding near Barrow, Alaska. Evidence of epibenthic feeding was based on mud visible on the dorsal surface of whales, resulting from feeding near the seafloor. Other cues used to assess feeding were an open mouth or the presence of feces in photographs. Over 3,600 photographs were analyzed including photos from surveys in spring BI 6727 order and late summer and in both the western and eastern Beaufort Sea. Of all the photographs analyzed, 64% were scored as definitively muddy. In spring, ratios ranged from a low of 27% in 2003 to a high of 76% in 2004. When all May sample sets off Barrow were combined (1985, 1986, 2003, 2004), there was a significant difference (t-test, P < 0.004) between the proportion of muddy juveniles to the proportion of muddy adults, with muddy adults being more common. The Barrow area was a commonly used feeding ground during migrations in both the spring (61% of the sample were feeding; 55% epibenthically) AZD6738 manufacturer and autumn (99% of the sample; 97% epibenthically). Bowheads both migrate and feed through areas where petroleum extraction is underway and anticipated; hence, exposure

to oil after a spill is of considerable concern to Native communities and management agencies. “
“Domoic acid (DA) is a neuroexcitatory toxin increasingly 上海皓元 causing strandings and mortality of marine mammals. The hippocampus of mammalian brains, associated with learning, memory, and spatial navigation, is one of the predominant regions affected by DA exposure. California sea lions stranding from 2003 to 2006 as a result of DA toxicosis were classified as having acute (n= 12) or chronic neurologic (n= 22) clinical signs. Chronic neurologic cases were examined by magnetic resonance imaging to determine the extent of brain damage related to DA exposure. Brain damage included hippocampal and parahippocampal

atrophy, temporal horn enlargement, and pathological T2 hyperintensity. Posttreatment, animals were fitted with satellite transmitters and their movement and dive behaviors compared with those of a control group. The only significant difference between acute and chronic animals was distance traveled per day. There were, however, significant differences between chronic neurologic cases and controls: chronic neurologic cases dove shallower for shorter durations, traveled further from shore, and spent less time hauled out and more time surface swimming than control animals. There was no relationship between severity of brain damage and behavioral patterns for chronic neurologic cases. Sea lions with chronic neurologic changes had a poor prognosis for survival following release.

This suggests that bowheads have a sense of smell, and we speculate that they may use this to find aggregations of krill on which they feed. “
“Aerial photographs were analyzed to investigate the feeding habits of the Bering-Chukchi-Beaufort (BCB) population of bowhead whales (Balaena mysticetus), particularly

epibenthic feeding near Barrow, Alaska. Evidence of epibenthic feeding was based on mud visible on the dorsal surface of whales, resulting from feeding near the seafloor. Other cues used to assess feeding were an open mouth or the presence of feces in photographs. Over 3,600 photographs were analyzed including photos from surveys in spring Saracatinib mw and late summer and in both the western and eastern Beaufort Sea. Of all the photographs analyzed, 64% were scored as definitively muddy. In spring, ratios ranged from a low of 27% in 2003 to a high of 76% in 2004. When all May sample sets off Barrow were combined (1985, 1986, 2003, 2004), there was a significant difference (t-test, P < 0.004) between the proportion of muddy juveniles to the proportion of muddy adults, with muddy adults being more common. The Barrow area was a commonly used feeding ground during migrations in both the spring (61% of the sample were feeding; 55% epibenthically) BVD-523 order and autumn (99% of the sample; 97% epibenthically). Bowheads both migrate and feed through areas where petroleum extraction is underway and anticipated; hence, exposure

to oil after a spill is of considerable concern to Native communities and management agencies. “
“Domoic acid (DA) is a neuroexcitatory toxin increasingly MCE公司 causing strandings and mortality of marine mammals. The hippocampus of mammalian brains, associated with learning, memory, and spatial navigation, is one of the predominant regions affected by DA exposure. California sea lions stranding from 2003 to 2006 as a result of DA toxicosis were classified as having acute (n= 12) or chronic neurologic (n= 22) clinical signs. Chronic neurologic cases were examined by magnetic resonance imaging to determine the extent of brain damage related to DA exposure. Brain damage included hippocampal and parahippocampal

atrophy, temporal horn enlargement, and pathological T2 hyperintensity. Posttreatment, animals were fitted with satellite transmitters and their movement and dive behaviors compared with those of a control group. The only significant difference between acute and chronic animals was distance traveled per day. There were, however, significant differences between chronic neurologic cases and controls: chronic neurologic cases dove shallower for shorter durations, traveled further from shore, and spent less time hauled out and more time surface swimming than control animals. There was no relationship between severity of brain damage and behavioral patterns for chronic neurologic cases. Sea lions with chronic neurologic changes had a poor prognosis for survival following release.

The second layer of regulation includes a series of modifications that regulate FOXO transcriptional activity by changing DNA binding and promoter binding specificity. This group includes acetylation

by the redox activated acetyl transferase, p300,[52-54] deacetylation by SIRT1,[55-57] SIRT2[58, 59] and SIRT3,[60] lysine methylation,[61, 62] and glycosylation.[20-22] Vemurafenib ic50 Lysine methylation at K270 of FOXO3 promotes loss of DNA binding and reduces FOXO-mediated apoptosis. Deacetylation by SIRT1 has been shown to differentially alter DNA binding affinity, so that more highly acetylated forms of FOXO3 favor expression of pro-apoptotic genes, (Bim, TRAIL, and FasL), while the more deacetylated forms favor expression of antioxidant and cytoprotective genes.[55] SIRT2 also deacetylates FOXOs and increases their DNA-binding activity.[58, 59] The binding of CBP/p300 to FOXOs is essential for transactivation of target genes.[52-54] However, the acetylation itself attenuates FOXO transcriptional activity. Several lysines were reported to be acetylated in FOXOs. Brunet et al. found that FOXO3 is acetylated at K242,K259, K271, K290, and K569 in the presence of stress stimuli.[55] Acetylation at K222, K245, K248,

K262, K265, K274, K294 of PD-0332991 nmr FOXO1 was also reported to regulate its DNA binding affinity and sensitivity to AKT phosphorylation.[63-65] Acetylation at K242, K245, and K262 of FOXO1 is sufficient to attenuate its transcriptional activity.[64] Fukuoka et al. reported the importance of K186, K189, and K408 deacetylation by HDAC in regulating FOXO4 transciptional activity.[66] O-glycosylation is another modification that

does not affect the nuclear/cytosolic distribution of FOXOs, but results in the upregulation of specific gene expression such as G6Pase[21] and other gluconeogenic genes.[20] Recent studies show that some of these effects involve the ability of specific PTMs, such as GlcNAcylation to produce differential binding of FOXOs to cofactors such as PGC-1α with a subsequent increase in specific transcriptional activities.[22] This second layer of modifications gives an idea of how FOXO transcriptional activity can be regulated. However, the question of how FOXOs decide which transcriptional program is activated in any given condition is still unclear. Since MCE公司 all FOXO proteins recognize a conserved consensus motif TTGTTTAC[67, 68] present in multiple genes, the promoter binding patterns may be defined more by differential binding to various cofactors. FOXOs have been shown to interact with a large number of binding partners resulting in changes in transcriptional activity of both proteins. The list includes a number of nuclear hormone receptors, other transcription factors such as β-catenin, runt-related transcription factor 3 (RUNX3), SMADs, and histone-modifying enzymes such as acetylases and methyltranferases (summarized by[69]).

The second layer of regulation includes a series of modifications that regulate FOXO transcriptional activity by changing DNA binding and promoter binding specificity. This group includes acetylation

by the redox activated acetyl transferase, p300,[52-54] deacetylation by SIRT1,[55-57] SIRT2[58, 59] and SIRT3,[60] lysine methylation,[61, 62] and glycosylation.[20-22] Idasanutlin cost Lysine methylation at K270 of FOXO3 promotes loss of DNA binding and reduces FOXO-mediated apoptosis. Deacetylation by SIRT1 has been shown to differentially alter DNA binding affinity, so that more highly acetylated forms of FOXO3 favor expression of pro-apoptotic genes, (Bim, TRAIL, and FasL), while the more deacetylated forms favor expression of antioxidant and cytoprotective genes.[55] SIRT2 also deacetylates FOXOs and increases their DNA-binding activity.[58, 59] The binding of CBP/p300 to FOXOs is essential for transactivation of target genes.[52-54] However, the acetylation itself attenuates FOXO transcriptional activity. Several lysines were reported to be acetylated in FOXOs. Brunet et al. found that FOXO3 is acetylated at K242,K259, K271, K290, and K569 in the presence of stress stimuli.[55] Acetylation at K222, K245, K248,

K262, K265, K274, K294 of BIBW2992 in vivo FOXO1 was also reported to regulate its DNA binding affinity and sensitivity to AKT phosphorylation.[63-65] Acetylation at K242, K245, and K262 of FOXO1 is sufficient to attenuate its transcriptional activity.[64] Fukuoka et al. reported the importance of K186, K189, and K408 deacetylation by HDAC in regulating FOXO4 transciptional activity.[66] O-glycosylation is another modification that

does not affect the nuclear/cytosolic distribution of FOXOs, but results in the upregulation of specific gene expression such as G6Pase[21] and other gluconeogenic genes.[20] Recent studies show that some of these effects involve the ability of specific PTMs, such as GlcNAcylation to produce differential binding of FOXOs to cofactors such as PGC-1α with a subsequent increase in specific transcriptional activities.[22] This second layer of modifications gives an idea of how FOXO transcriptional activity can be regulated. However, the question of how FOXOs decide which transcriptional program is activated in any given condition is still unclear. Since MCE all FOXO proteins recognize a conserved consensus motif TTGTTTAC[67, 68] present in multiple genes, the promoter binding patterns may be defined more by differential binding to various cofactors. FOXOs have been shown to interact with a large number of binding partners resulting in changes in transcriptional activity of both proteins. The list includes a number of nuclear hormone receptors, other transcription factors such as β-catenin, runt-related transcription factor 3 (RUNX3), SMADs, and histone-modifying enzymes such as acetylases and methyltranferases (summarized by[69]).

As shown in Fig. 4A, we observed robust up-regulation of the BH3-only protein, Bnip3, by GANT61 in all three HCC cell lines; the expression of other Bcl-2 family proteins (including Bim, Noxa, Puma, Bcl2, and Bclxl) were not significantly affected, although the level of Mcl-1 was selleckchem slightly reduced in two of the three HCC cell lines. GANT61 induced a 4.39-fold, 2.84-fold, and 1.97-fold increase in Bnip3 mRNA level in Huh7, Hep3B, and HepG2 cells, respectively, as determined by qRT-PCR (Fig. 4B). The effect of GANT61 on Bnip3 expression was dose-dependent (at 24- and 48-hour timepoints) (Fig. 4C). The role of Bnip3 in GANT61-induced autophagy was supported by the observations that siRNA knockdown

of Bnip3 prevented GANT61-induced LC3II accumulation (Fig. 4D, left panel) and that overexpression of Bnip3 enhanced GANT61-induced LC3II accumulation (Fig. 4D, right panel) and reversed SAG-induced LC3II reduction (Fig. 4E). We sought to further investigate the mechanism by which Hh signaling regulates Bnip3. As the Bnip3 promoter does not contain the Gli consensus DNA-binding sequences, it is likely that Hh signaling might regulate Bnip3 through an indirect mechanism. Given that Bnip3 is a downstream target of the MEK/ERK signaling pathway[11, 12]

and that Hh and MEK/ERK signaling pathways are known to interconnect in other cells,[13-15] we performed experiments DNA Synthesis inhibitor to determine whether inhibition of Hh by GANT61 might induce Bnip3 expression by way of activation of MEK/ERK. As shown in Fig. 4F, GANT61 treatment increased the phosphorylation of MEK and ERK1/2

(but did not 上海皓元 affect the levels of total MEK and ERK1/2). We observed that inhibition of MEK by U0126 prevented GANT61-induced phosphorylation of ERK1/2, expression of Bnip3, and accumulation of LC3II (Fig. 4G). These findings suggest that GANT61-induced Bnip3 expression is mediated at least in part through activation of the MEK/ERK pathway. Although Bnip expression is known to be regulated by nuclear factor kappa B (NF-κB),[16] p53,[17] and DNA methyltransferase-1 (DNMT-1),[18] these molecules were not altered by GANT61 treatment in our system (Supporting Fig. S3). Beclin-1, the mammalian ortholog of yeast Atg6, is a well-known key regulator of autophagy; it is a critical component of the class III phosphatidylinositol-3-kinase complex (PI3KC3) required for autophagy. The overall structure of Beclin-1/Atg6 and its essential role in autophagosome formation is evolutionarily conserved throughout all eukaryotic phyla. Whereas Beclin-1 expression promotes autophagy, Beclin-1 reduction decreases autophagic activity.[19] Sequence alignment and structural modeling indicate that Beclin-1 contains a putative BH3-like domain (amino acids 112-123), which is known as a novel BH-3 domain only protein.[20] The BH-3 domain of Beclin-1 interacts with Bcl-2, which leads to inhibition of autophagy.