_:b549506998 . . . . _:b549506993 . . _:b549507083 . _:b549506992 . _:b549506990 "6"^^ . _:b549506995 . _:b549506994 . _:b549506991 "5"^^ . _:b549507005 . _:b30247475 _:b30247496 . _:b30247475 _:b30247497 . _:b549507004 . _:b30247475 _:b30247498 . _:b30247475 _:b30247499 . _:b30247495 "Angiogenesis is required for invasive tumor growth and metastasis. Therefore, it constitutes an important point in the control of cancer progression and a target to inhibit further growth.>>47<< Unlike normal blood vessels, the tumor vasculature is abnormally composed, resulting in the formation of physiological barriers which hamper the delivery of therapeutic agents into tumors." . _:b30247475 _:b30247500 . _:b30247475 _:b30247501 . _:b549507007 . _:b30247475 _:b30247502 . _:b30247475 _:b30247503 . _:b30247475 _:b30247488 . _:b30247475 _:b30247489 . _:b549507032 . _:b30247475 _:b30247490 . _:b549507006 . _:b30247478 . _:b30247475 _:b30247491 . _:b30247475 _:b30247492 . . _:b30247475 _:b30247493 . _:b549507001 . _:b30247475 _:b30247494 . _:b30247475 _:b30247495 . . _:b549507000 . _:b30247488 "However, a combination of free cisplatin and RAPA resulted in a much lower IC50 of 0.82 \u03BCM (based on the concentration of cisplatin) (Figure 3, A). A combination index (CI) was determined using the Chou\u2013Talalay isobologram equation.13,>>14<< The CI at the IC50 of the free drug combination was 0.36, indicating strong synergism." . _:b549507006 . _:b549506997 "4"^^ . _:b549507003 . _:b30247475 _:b30247504 . _:b549506996 "4"^^ . _:b30247475 _:b30247505 . _:b549507002 . _:b30247475 _:b30247506 . _:b30247475 _:b30247507 . _:b30247475 _:b30247508 . . _:b30247475 _:b30247509 . _:b549506999 "4"^^ . _:b549507013 . . _:b549506998 "4"^^ . . _:b30247508 "Previously, our laboratory has successfully encapsulated drugs such as gemcitabine triphosphate,>>22<< siRNA, DNA,23 peptide,25 and other types of hydrophilic phosphorated drugs into DOPA-coated calcium phosphate cores." . _:b549507012 . _:b549506993 "4"^^ . _:b549507015 . . _:b549506992 "5"^^ . _:b549507014 . . _:b30247482 . _:b549506995 "4"^^ . _:b549507009 . _:b549507101 . . _:b549506994 "4"^^ . _:b549507008 . . _:b549507005 "3"^^ . . _:b549507011 . _:b549507004 "3"^^ . _:b549507040 . _:b549507010 . _:b549507007 "3"^^ . . _:b549507021 . . _:b549507006 "3"^^ . _:b549507012 . . _:b549507020 . _:b549507055 . _:b549507001 "3"^^ . _:b549507023 . _:b549507000 "3"^^ . _:b549507022 . _:b30247483 "receptors are well-known membrane-bound proteins that show high affinity for neuroleptics and are overexpressed on many human tumors including melanoma, non-small cell lung carcinoma, breast tumors of neural origin, and prostate cancer.>>31<<\u221233 Sigma receptors bind to haloperidol and various other neuroleptics showing a high affinity with the anisamide moiety." . _:b549507003 "3"^^ . _:b549507017 . _:b549507002 "3"^^ . _:b30247489 "that RAPA significantly sensitized A375-luc melanoma cells to apoptosis induced by cisplatin, possibly because mTOR inhibitors sensitize tumor cells to cisplatin by blocking the up-regulation of p21 and inducing apoptosis as a result.>>19<<" . _:b549507016 . _:b549507013 "3"^^ . _:b549507019 . _:b549507012 "3"^^ . _:b549507100 . . _:b549507018 . _:b549507015 "3"^^ . _:b549507029 . _:b30247474 "DOPA-coated cisplatin cores were prepared according to our previous publications,>>25<<,26 dispersed in THF, and stored in a glass vial for further use." . . _:b549507014 "3"^^ . _:b549507028 . _:b549507009 "3"^^ . _:b549507031 . _:b549507008 "3"^^ . _:b549507029 . _:b549507030 . . _:b549507011 "3"^^ . . _:b549507025 . _:b549507010 "3"^^ . _:b549507073 . _:b549507024 . _:b549507021 "2"^^ . _:b549507027 . _:b549507020 "2"^^ . _:b549507026 . _:b30247503 . _:b549507023 "2"^^ . _:b30247502 . _:b549507037 . _:b549507022 "2"^^ . . _:b549507072 . . _:b549507036 . . _:b30247497 . _:b549507017 "3"^^ . _:b549507039 . _:b549507016 "3"^^ . _:b30247496 . _:b549507038 . _:b549507019 "2"^^ . _:b549507023 . _:b549507033 . _:b549507018 "3"^^ . . . _:b549507032 . _:b549507029 "2"^^ . _:b30247505 "composed of the surrounding blood vessels, immune cells, fibroblasts, pericytes, endothelial cells, signaling molecules, and the extracellular matrix,53 and many important molecular targets in the microenvironment have been identified.>>54<< Therefore, the tumor microenvironment has become an important aim for maximizing the effects of chemotherapy through combination therapy." . . _:b549507035 . _:b549507028 "2"^^ . . _:b549506996 . _:b549507034 . _:b549507031 "2"^^ . _:b549507045 . _:b549507030 "2"^^ . _:b30247509 . _:b30247485 "It has been reported that RAPA is capable of sensitizing melanoma cells to cisplatin.>>19<<\u221221 PLGA NPs loaded with a 4.5 wt % of cisplatin cores and 2.2 wt % of RAPA corresponding to a cisplatin/RAPA molar ratio of 5.5:" . _:b549507044 . _:b549507025 "2"^^ . _:b549507047 . _:b549507024 "2"^^ . _:b549507046 . _:b549507076 . _:b549507000 . _:b549507027 "2"^^ . _:b549507063 . _:b549507041 . _:b549507026 "2"^^ . _:b30247509 "Previously, our laboratory has successfully encapsulated drugs such as gemcitabine triphosphate,22 siRNA, DNA,23 peptide,>>25<< and other types of hydrophilic phosphorated drugs into DOPA-coated calcium phosphate cores." . _:b549507040 . _:b549507037 "2"^^ . _:b30247492 "It was also found that liver was the major organ to clear PLGA NPs, which is consistent with the biodistribution of other PLGA NPs.>>44<<" . _:b549507043 . _:b549507036 "2"^^ . _:b549507053 . . . _:b549507042 . _:b549507039 "2"^^ . . _:b549507053 . _:b549507098 . _:b549507038 "2"^^ . _:b30247494 "As collagen is secreted by TAFs (tumor-associated fibroblasts), the effect of this treatment on TAFs was further investigated by staining for \u03B1-smooth muscle actin (SMA), a marker of TAFs.>>46<< Apoptosis was confirmed through terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) (Figure 8)." . _:b549507052 . . _:b549507033 "2"^^ . . . . _:b549507055 . _:b549507032 "2"^^ . . _:b30247491 . _:b549507068 . . _:b549507054 . . . _:b549507035 "2"^^ . _:b549507049 . _:b549507034 "2"^^ . _:b549507048 . _:b549507064 . _:b549507045 "2"^^ . _:b549507051 . _:b549507044 "2"^^ . . _:b549507050 . _:b549507047 "2"^^ . . _:b549507061 . _:b549507024 . . _:b549507046 "2"^^ . _:b549507060 . _:b549507080 . _:b549507041 "2"^^ . _:b549507063 . _:b549507040 "2"^^ . _:b549507010 . _:b549507062 . _:b549507043 "2"^^ . _:b549507057 . . _:b549507042 "2"^^ . _:b549507025 . . _:b549507056 . _:b549507053 "2"^^ . . _:b549507059 . _:b549507052 "2"^^ . _:b549507058 . _:b549507055 "2"^^ . _:b549507069 . _:b549507054 "2"^^ . _:b549507068 . _:b549506997 . _:b549507049 "2"^^ . _:b549507071 . _:b549507048 "2"^^ . _:b549507070 . _:b549507051 "2"^^ . _:b30247474 . _:b549507037 . _:b549507065 . . _:b549507050 "2"^^ . _:b549507064 . _:b549507061 "2"^^ . _:b549507067 . _:b549507060 "2"^^ . . _:b549507066 . _:b549507063 "2"^^ . . _:b549507062 "2"^^ . _:b30247477 . _:b549507057 "2"^^ . . _:b549507056 "2"^^ . _:b549506990 . _:b549506991 . _:b549506992 . _:b549506993 . _:b549507059 "2"^^ . _:b549506994 . _:b549506995 . _:b549506996 . _:b549506997 . _:b549507058 "2"^^ . _:b549506998 . _:b549506999 . _:b549507000 . _:b549507001 . _:b549507069 "2"^^ . _:b549507002 . _:b549507003 . _:b549507087 . _:b549507004 . _:b549507056 . _:b549507005 . _:b549507068 "2"^^ . _:b549507006 . _:b549507007 . _:b549507043 . _:b549507008 . _:b549507009 . _:b549507071 "2"^^ . _:b549507010 . _:b549507011 . _:b549507012 . _:b549507013 . _:b549507014 . _:b549507070 "2"^^ . _:b549507015 . _:b549507016 . _:b549507017 . _:b30247479 . _:b30247480 . _:b549507008 . _:b549507018 . _:b549507065 "2"^^ . . _:b549507019 . . _:b549507020 . _:b549507021 . _:b549507022 . _:b549507064 "2"^^ . _:b549507023 . _:b549507024 . _:b549507025 . _:b549507026 . _:b549507067 "2"^^ . _:b549507027 . _:b549507028 . _:b549507001 . _:b549507029 . _:b549507030 . _:b549507066 "2"^^ . _:b549507031 . _:b549507032 . _:b549507033 . _:b549507034 . _:b549507035 . _:b549507036 . _:b549507037 . _:b549507038 . _:b549507039 . _:b549507040 . _:b549507041 . _:b549507042 . _:b549507043 . _:b549507089 . _:b549507044 . . _:b549507045 . _:b549507046 . _:b549507047 . . _:b549507048 . _:b549507049 . . _:b549507050 . _:b549507051 . _:b549507052 . _:b549507053 . _:b30247488 . _:b549507054 . _:b549507055 . _:b30247476 "Cisplatin is a first-line therapy against a wide spectrum of solid neoplasms. However, systemic toxicities including nephro- and neurotoxicities along with drug resistance severely limit the use of cisplatin in the clinical setting.34\u2212>>36<< In an effort to overcome the associated nephro- and neurotoxicities, attempts to encapsulate cisplatin into NPs were made." . _:b549507056 . _:b549507057 . _:b549507058 . _:b549507059 . _:b549507060 . _:b549507061 . _:b549507062 . _:b549507063 . _:b549507064 . _:b549507065 . _:b549507066 . _:b549507067 . _:b549507068 . _:b549507069 . _:b549507060 . _:b549507070 . _:b549507071 . . _:b30247498 . . _:b30247484 "are well-known membrane-bound proteins that show high affinity for neuroleptics and are overexpressed on many human tumors including melanoma, non-small cell lung carcinoma, breast tumors of neural origin, and prostate cancer.31\u2212>>33<< Sigma receptors bind to haloperidol and various other neuroleptics showing a high affinity with the anisamide moiety." . _:b549507075 . _:b30247490 "that RAPA significantly sensitized A375-luc melanoma cells to apoptosis induced by cisplatin, possibly because mTOR inhibitors sensitize tumor cells to cisplatin by blocking the up-regulation of p21 and inducing apoptosis as a result.19\u2212>>21<<" . . . . _:b30247505 . _:b30247508 . . . _:b549507051 . . . . . _:b30247507 "through combination therapy. For example, recent studies have shown that the administration of low dose TGF-\u03B2 inhibitor significantly decreases the IFP in the tumor and therefore enhances the accumulation and penetration of NPs.55,>>56<< However, the hydrophobicity of the TGF-\u03B2 inhibitor limits its use and the encapsulation multiple drugs into NPs with diverse physicochemical properties has previously proven difficult." . _:b549507054 . . _:b549507026 . _:b549507048 . _:b549506991 . _:b549507017 . . _:b549507102 . . _:b549507016 . _:b549507007 . _:b30247492 . _:b549507058 . _:b549506994 . . _:b549507038 . _:b30247484 . . _:b549507042 . . _:b549507091 . . _:b549507094 . _:b549507071 . _:b549507052 . . _:b549507062 . _:b30247507 . _:b549507077 . . _:b30247481 "% and the EE of RAPA was still facilitated by 2.8-fold in the presence of the cisplatin cores. This finding is consistent with previous observations that oleic acid coated NPs can enhance encapsulation of hydrophobic drugs in PLGA NP.40\u2212>>42<< DLS results confirmed that the size and monodispersity of all PLGA NPs was not affected by the encapsulation of RAPA, cisplatin cores, or both drugs in a single NP (Figure S1, Supporting Information)." . . . _:b30247476 . _:b30247477 . _:b30247478 . _:b549507074 . _:b549507070 . _:b549507005 . _:b30247479 . . _:b30247498 "in the tumor in mice treated with cisplatin PLGA NPs alone was only slightly affected by the treatment. In addition, blood vessel normalization was observed with RAPA treatment groups, consistent with previous reports in breast cancers.>>17<<,45,49 Normalization may enhance transvascular flux and improve the delivery of NPs to the tumor." . _:b30247473 . _:b30247474 . _:b30247475 . . _:b30247484 . _:b549507092 . _:b30247485 . . _:b549507082 . _:b30247486 . _:b30247487 . _:b30247485 . _:b549507033 . _:b30247480 . _:b30247481 . . . _:b30247482 . _:b30247477 "drug loading through a reverse microemulsion method.26 The resulting cisplatin cores are composed entirely of cisplatin and stabilized by DOPA with a drug loading of approximately 93 wt % and have tunable sizes between 12 and 75 nm.>>25<<,26 After being coated with an outer leaflet lipid layer, cisplatin cores demonstrated potent antitumor effects both in vivo and in vitro." . _:b30247483 . . . _:b30247492 . . _:b30247493 . . . _:b549507034 . _:b30247494 . . . _:b549507086 . . _:b30247495 . . _:b549507065 . _:b30247488 . . _:b30247489 . _:b549506990 . _:b30247490 . _:b30247504 "The tumor microenvironment is composed of the surrounding blood vessels, immune cells, fibroblasts, pericytes, endothelial cells, signaling molecules, and the extracellular matrix,>>53<< and many important molecular targets in the microenvironment have been identified." . . _:b549507078 . . _:b30247491 . _:b549507046 . _:b549507090 . _:b30247500 . . _:b549507003 . . _:b30247501 . . . _:b549507079 . . _:b30247502 . _:b30247503 . _:b30247496 . _:b30247497 . _:b30247498 . _:b30247473 "materials and methods" . _:b30247499 . _:b549507103 . _:b30247508 . . _:b549507097 . _:b30247509 . . _:b30247475 "results and discussion" . _:b30247499 . . _:b30247504 . . _:b30247505 . _:b30247506 . . . _:b30247507 . . _:b549507067 . . _:b30247501 . "10.1021%2Fnn5010815" . . _:b549507077 . _:b30247495 . _:b30247503 "penetration of gemcitabine inside the pancreatic tumor. Preclinical data have also shown that PEGylated Cellax reduced the content of \u03B1-smooth muscle actin, which is a marker of TAFs, in 4T1 and MDA-MB-231 orthotopic breast tumor models.>>5<< Cellax treatment significantly decreased the IFP of tumor. As a result, the perfusion of active drugs into tumors was significantly enhanced, resulting in a notable tumor growth inhibition." . _:b549506998 . _:b549507076 . _:b549507022 . _:b549507079 . _:b549507078 . _:b30247479 "Therefore, the encapsulation is limited by the compatibility between RAPA and the hydrophobic block copolymer.37\u2212>>39<< However, when RAPA was coencapsulated alongside 4.5 wt % cisplatin cores at a 5.0 wt % feed ratio (combinatory PLGA NPs), the EE and DL of RAPA in combinatory PLGA NPs were subsequently increased to 80% and 4.0 wt % (Figure 1D)," . _:b549507073 . . _:b30247493 . _:b549507072 . _:b549507075 . _:b30247476 . _:b30247490 . _:b549507074 . _:b549507057 . . _:b549507085 . _:b549507084 . _:b549507087 . _:b549507035 . _:b549507086 . . _:b549507081 . _:b549507085 . _:b549507080 . _:b30247494 . _:b549507083 . _:b549507082 . _:b549507093 . _:b549507049 . _:b549507077 "2"^^ . . _:b549507092 . _:b549507076 "2"^^ . . _:b549507095 . _:b549507079 "2"^^ . . _:b549507094 . _:b549507078 "2"^^ . _:b549507089 . _:b549507028 . _:b549507073 "2"^^ . . _:b549507088 . _:b549507072 "2"^^ . _:b549507045 . _:b549507091 . . _:b549507075 "2"^^ . . _:b549507090 . _:b549507009 . _:b549506995 . _:b30247480 "wt % and the EE of RAPA was still facilitated by 2.8-fold in the presence of the cisplatin cores. This finding is consistent with previous observations that oleic acid coated NPs can enhance encapsulation of hydrophobic drugs in PLGA NP.>>40<<\u221242 DLS results confirmed that the size and monodispersity of all PLGA NPs was not affected by the encapsulation of RAPA, cisplatin cores, or both drugs in a single NP (Figure S1, Supporting Information)." . _:b549507044 . _:b549507074 "2"^^ . _:b549507101 . _:b549507085 "2"^^ . _:b549507100 . _:b549507027 . . _:b549507084 "2"^^ . _:b549507103 . _:b549507087 "2"^^ . _:b549507102 . _:b549507086 "2"^^ . . _:b549507097 . _:b549507081 "2"^^ . _:b549507096 . . _:b549507080 "2"^^ . _:b549507099 . _:b549507083 "2"^^ . _:b549507098 . . _:b549507082 "2"^^ . _:b549507093 "2"^^ . _:b549507018 . _:b549507092 "2"^^ . _:b30247486 . _:b549507002 . _:b549507095 "2"^^ . _:b549507094 "2"^^ . . _:b549507089 "2"^^ . . _:b30247501 "An elevated IFP, a physical barrier from the extra cellular matrix, hinders the delivery of drugs into the dense collagen matrix.>>45<< Our studies have shown that combinatory PLGA NPs modulated the stroma by depleting TAFs and decreasing the amount of collagen." . _:b549507088 "2"^^ . _:b549507066 . _:b549507091 "2"^^ . _:b549507090 "2"^^ . _:b549507014 . . _:b549507101 "2"^^ . _:b549507041 . _:b549507100 "2"^^ . . _:b549507103 "2"^^ . _:b549507039 . . _:b549507102 "2"^^ . _:b30247500 "tumor in mice treated with cisplatin PLGA NPs alone was only slightly affected by the treatment. In addition, blood vessel normalization was observed with RAPA treatment groups, consistent with previous reports in breast cancers.17,45,>>49<< Normalization may enhance transvascular flux and improve the delivery of NPs to the tumor." . _:b549507097 "2"^^ . _:b549507059 . _:b549507095 . _:b549507096 "2"^^ . . _:b549506992 . _:b549507099 "2"^^ . _:b30247482 "Co-encapsulation of multiple agents with diverse physicochemical properties, such as calcium phosphate cores loaded with gemcitabine and macromolecular siRNA, may be possible.>>22<<" . . _:b549507098 "2"^^ . . _:b549507004 . _:b549507099 . . . . _:b549507031 . _:b549507015 . _:b549507093 . _:b549507011 . _:b30247481 . _:b549507072 . _:b549507073 . _:b549507074 . _:b549507075 . . _:b549507076 . _:b549507077 . _:b549507078 . _:b549507079 . _:b549507080 . _:b549507081 . _:b549507082 . . _:b549507083 . _:b549507084 . _:b549507085 . _:b30247506 . _:b549507086 . _:b549507087 . _:b549507088 . _:b549507089 . _:b549507090 . _:b549507091 . _:b549507092 . _:b549507093 . _:b549507094 . _:b549507095 . _:b549507096 . _:b549507061 . _:b549507097 . _:b549507098 . _:b549507030 . . _:b549507099 . _:b549507100 . _:b549507101 . _:b30247506 "through combination therapy. For example, recent studies have shown that the administration of low dose TGF-\u03B2 inhibitor significantly decreases the IFP in the tumor and therefore enhances the accumulation and penetration of NPs.>>55<<,56 However, the hydrophobicity of the TGF-\u03B2 inhibitor limits its use and the encapsulation multiple drugs into NPs with diverse physicochemical properties has previously proven difficult." . _:b30247497 "barriers which hamper the delivery of therapeutic agents into tumors.48 Therefore, antiangiogenic imbalances in the tumor results in tumor vasculature normalization and thus improves NP delivery and accumulation in the tumor.>>48<< Combination of cytotoxic therapies and antiangiogenic treatment during the vascular normalization also exhibits synergistic effects." . _:b549507102 . _:b549507103 . _:b30247486 "It has been reported that RAPA is capable of sensitizing melanoma cells to cisplatin.19\u2212>>21<< PLGA NPs loaded with a 4.5 wt % of cisplatin cores and 2.2 wt % of RAPA corresponding to a cisplatin/RAPA molar ratio of 5.5:" . _:b549507081 . _:b30247487 "However, a combination of free cisplatin and RAPA resulted in a much lower IC50 of 0.82 \u03BCM (based on the concentration of cisplatin) (Figure 3, A). A combination index (CI) was determined using the Chou\u2013Talalay isobologram equation.>>13<<,14 The CI at the IC50 of the free drug combination was 0.36, indicating strong synergism." . _:b549507084 . _:b30247499 "the tumor in mice treated with cisplatin PLGA NPs alone was only slightly affected by the treatment. In addition, blood vessel normalization was observed with RAPA treatment groups, consistent with previous reports in breast cancers.17,>>45<<,49 Normalization may enhance transvascular flux and improve the delivery of NPs to the tumor." . _:b30247504 . . _:b30247473 . _:b30247487 . _:b30247475 . . . "PMC0" . _:b30247478 "Therefore, the encapsulation is limited by the compatibility between RAPA and the hydrophobic block copolymer.>>37<<\u221239 However, when RAPA was coencapsulated alongside 4.5 wt % cisplatin cores at a 5.0 wt % feed ratio (combinatory PLGA NPs), the EE and DL of RAPA in combinatory PLGA NPs were subsequently increased to 80% and 4.0 wt % (Figure 1D)," . _:b30247493 "A dense extracellular matrix (ECM) and the stromal components of tumors contribute to an increased interstitial fluid pressure (IFP), which severely hampers efficient penetration of NPs into the tumor.>>45<< As a result, cells distal from blood vessels are resistant to systemic therapy because of low drug availability." . . _:b30247500 . _:b549507069 . _:b549506999 . . . _:b549507096 . _:b30247475 _:b30247480 . _:b30247483 . _:b30247475 _:b30247481 . . _:b30247473 _:b30247474 . _:b30247475 _:b30247482 . _:b30247475 _:b30247483 . _:b549507036 . _:b30247475 _:b30247484 . _:b30247475 _:b30247485 . _:b30247502 "through both clinical and research data, weakening the tumor stroma has proven successful in enhancing the efficacy of chemotherapy. For example, abraxane was combined with gemcitabine to treat pancreatic cancer in a phase I/II study.50,>>51<< The combination showed significant anticancer efficacy with a 48% response rate, which was attributed to the depletion of stroma for enhanced penetration of gemcitabine inside the pancreatic tumor." . . _:b30247475 _:b30247486 . _:b30247475 _:b30247487 . . _:b30247489 . _:b30247475 _:b30247476 . _:b30247475 _:b30247477 . _:b30247475 _:b30247478 . _:b30247475 _:b30247479 . _:b30247496 "and a target to inhibit further growth.47 Unlike normal blood vessels, the tumor vasculature is abnormally composed, resulting in the formation of physiological barriers which hamper the delivery of therapeutic agents into tumors.>>48<< Therefore, antiangiogenic imbalances in the tumor results in tumor vasculature normalization and thus improves NP delivery and accumulation in the tumor." . _:b549506991 . _:b549507019 . _:b549506990 . _:b549507013 . _:b549507047 . _:b549507050 . _:b549507020 . . _:b30247491 "from combinatory PLGA NPs during the circulation. Eight hours post-iv injection, 6.0% of the injected dose per gram of cisplatin in combinatory NPs accumulated in the tumor (Figure 5, B), which was significantly higher than free CDDP.>>25<< Meanwhile, the accumulation ratio of 14C-RAPA/cisplatin/3H-labeled combinatory NPs was near 1:" . _:b549507021 . _:b549506993 . _:b549507088 . _:b549506997 . . . _:b549506996 . . _:b549506999 .