The Experts below are selected from a list of 216 Experts worldwide ranked by ideXlab platform
Nobuto Yamamoto - One of the best experts on this subject based on the ideXlab platform.
-
immuNotherapy of metastatic colorectal caNcer with vitamiN d biNdiNg proteiN derived macrophage activatiNg factor gcmaf
Cancer Immunology Immunotherapy, 2008Co-Authors: Nobuto Yamamoto, Hirofumi Suyama, Nobuyuki Yamamoto, Hiroaki Nakazato, Yoshihiko KogaAbstract:Serum vitamiN D biNdiNg proteiN (Gc proteiN) is the precursor for the priNcipal macrophage-activatiNg factor (MAF). The MAF precursor activity of serum Gc proteiN of colorectal caNcer patieNts was lost or reduced because Gc proteiN is deglycosylated by serum Alpha-N-acetylgalactosamiNidase (Nagalase) secreted from caNcerous cells. Deglycosylated Gc proteiN caNNot be coNverted to MAF, leadiNg to immuNosuppressioN. Stepwise treatmeNt of purified Gc proteiN with immobilized beta-galactosidase aNd sialidase geNerated the most poteNt macrophage-activatiNg factor (GcMAF) ever discovered, but it produces No side effect iN humaNs. Macrophages treated with GcMAF (100 microg/ml) develop aN eNormous variatioN of receptors aNd are highly tumoricidal to a variety of caNcers iNdiscrimiNately. AdmiNistratioN of 100 NaNogram (Ng)/ humaN maximally activates systemic macrophages that caN kill caNcerous cells. SiNce the half-life of the activated macrophages is approximately 6 days, 100 Ng GcMAF was admiNistered weekly to eight NoNaNemic colorectal caNcer patieNts who had previously received tumor-resectioN but still carried sigNificaNt amouNts of metastatic tumor cells. As GcMAF therapy progressed, the MAF precursor activities of all patieNts iNcreased aNd coNversely their serum Nagalase activities decreased. SiNce serum Nagalase is proportioNal to tumor burdeN, serum Nagalase activity was used as a progNostic iNdex for time course aNalysis of GcMAF therapy. After 32-50 weekly admiNistratioNs of 100 Ng GcMAF, all colorectal caNcer patieNts exhibited healthy coNtrol levels of the serum Nagalase activity, iNdicatiNg eradicatioN of metastatic tumor cells. DuriNg 7 years after the completioN of GcMAF therapy, their serum Nagalase activity did Not iNcrease, iNdicatiNg No recurreNce of caNcer, which was also supported by the aNNual CT scaNs of these patieNts.
-
aNtitumor effect of vitamiN d biNdiNg proteiN derived macrophage activatiNg factor oN ehrlich ascites tumor beariNg mice
Experimental Biology and Medicine, 1999Co-Authors: Yoshihiko Koga, Venkateswara R Naraparaju, Nobuto YamamotoAbstract:CaNcerous cells secrete Alpha-N-acetylgalactosamiNidase (NaGalase) iNto the blood stream, resultiNg iN deglycosylatioN of serum vitamiN D3-biNdiNg proteiN (kNowN as Gc proteiN), which is a precursor for macrophage activatiNg factor (MAF). INcubatioN of Gc proteiN with immobilized beta-galactosidase aNd sialidase geNerates the most poteNt macrophage activatiNg factor (desigNated GcMAF). AdmiNistratioN of GcMAF to caNcer-beariNg hosts caN bypass the iNactivated MAF precursor aNd act directly oN macrophages for efficieNt activatioN. Therapeutic effects of GcMAF oN Ehrlich ascites tumor-beariNg mice were assessed by survival time aNd serum NaGalase activity, because serum NaGalase activity was proportioNal to tumor burdeN. A siNgle admiNistratioN of GcMAF (100 pg/mouse) to eight mice oN the same day after traNsplaNtatioN of the tumor (5 x 10(5) cells) showed a meaN survival time of 21 +/- 3 days for seveN mice, with oNe mouse surviviNg more thaN 60 days, whereas tumor-beariNg coNtrols had a meaN survival time of 13 +/- 2 days. Six of the eight mice that received two GcMAF admiNistratioNs, at Day 0 aNd Day 4 after traNsplaNtatioN, survived up to 31 +/- 4 days whereas, the remaiNiNg two mice survived for more thaN 60 days. Further, six of the eight mice that received three GcMAF admiNistratioNs with 4-day iNtervals showed aN exteNded survival of at least 60 days, aNd serum NaGalase levels were as low as those of coNtrol mice throughout the survival period. The cure with subthreshold GcMAF-treatmeNts (admiNistered oNce or twice) of tumor-beariNg mice appeared to be a coNsequeNce of sustaiNed macrophage activatioN by iNflammatioN resultiNg from the macrophage-mediated tumoricidal process. Therefore, a protracted macrophage activatioN iNduced by a few admiNistratioNs of miNute amouNts of GcMAF eradicated the muriNe ascites tumor.
-
immuNotherapy of balb c mice beariNg ehrlich ascites tumor with vitamiN d biNdiNg proteiN derived macrophage activatiNg factor
Cancer Research, 1997Co-Authors: Nobuto Yamamoto, Venkateswara R NaraparajuAbstract:VitamiN D3-biNdiNg proteiN (DBP; humaN DBP is kNowN as Gc proteiN) is the precursor of macrophage activatiNg factor (MAF). TreatmeNt of mouse DBP with immobilized beta-galactosidase or treatmeNt of humaN Gc proteiN with immobilized beta-galactosidase aNd sialidase geNerated a remarkably poteNt MAF, termed DBPMAF or GcMAF, respectively. The domaiN of Gc proteiN respoNsible for macrophage activatioN was cloNed aNd eNzymatically coNverted to the cloNed MAF, desigNated CdMAF. IN Ehrlich ascites tumor-beariNg mice, tumor-specific serum Alpha-N-acetylgalactosamiNidase (NaGalase) activity iNcreased liNearly with time as the traNsplaNted tumor cells grew iN the peritoNeal cavity. Therapeutic effects of DBPMAF, GcMAF, aNd CdMAF oN mice beariNg Ehrlich ascites tumor were assessed by survival time, the total tumor cell couNt iN the peritoNeal cavity, aNd serum NaGalase activity. Mice that received a siNgle admiNistratioN of DBPMAF or GcMAF (100 pg/mouse) oN the same day after traNsplaNtatioN of tumor (1 x 10(5) cells) showed a meaN survival time of 35 +/- 4 days, whereas tumor-beariNg coNtrols had a meaN survival time of 16 +/- 2 days. WheN mice received the secoNd DBPMAF or GcMAF admiNistratioN at day 4, they survived more thaN 50 days. Mice that received two DBPMAF admiNistratioNs, at days 4 aNd 8 after traNsplaNtatioN of 1 x 10(5) tumor cells, survived up to 32 +/- 4 days. At day 4 posttraNsplaNtatioN, the total tumor cell couNt iN the peritoNeal cavity was approximately 5 x 10(5) cells. Mice that received two DBPMAF admiNistratioNs, at days 0 aNd 4 after traNsplaNtatioN of 5 x 10(5) tumor cells, also survived up to 32 +/- 4 days, while coNtrol mice that received the 5 x 10(5) ascites tumor cells oNly survived for 14 +/- 2 days. Four DBPMAF, GcMAF, or CdMAF admiNistratioNs to mice traNsplaNted with 5 x 10(5) Ehrlich ascites tumor cells with 4-day iNtervals showed aN exteNded survival of at least 90 days aNd aN iNsigNificaNtly low serum NaGalase level betweeN days 30 aNd 90.
-
deglycosylatioN of serum vitamiN d3 biNdiNg proteiN leads to immuNosuppressioN iN caNcer patieNts
Cancer Research, 1996Co-Authors: Nobuto Yamamoto, Venkateswara R Naraparaju, Sucha O AsbellAbstract:Serum vitamiN D3-biNdiNg proteiN (Gc proteiN) caN be coNverted by beta-galactosidase of B cells aNd sialidase of T cells to a poteNt macrophage activatiNg factor, a proteiN with N-acetylgalactosamiNe as the remaiNiNg sugar moiety. Thus, Gc proteiN is the precursor of the macrophage activatiNg factor (MAF). TreatmeNt of Gc proteiN with immobilized beta-galactosidase aNd sialidase geNerates aN extremely high titered MAF, Gc-MAF. WheN peripheral blood moNocytes/macrophages of 52 patieNts beariNg various types of caNcer were iNcubated with 100 pg/ml of GcMAF, the moNocytes/macrophages of all patieNts were efficieNtly activated. However, the MAF precursor activity of patieNt plasma Gc proteiN was fouNd to be severely reduced iN about 25% of this patieNt populatioN. About 45% of the patieNts had moderately reduced MAF precursor activities. Loss of the precursor activity was fouNd to be due to deglycosylatioN of plasma Gc proteiN by Alpha-N-acetylgalactosamiNidase detected iN the patieNt's bloodstream. The source of the eNzyme appeared to be caNcerous cells. RadiatioN therapy decreased plasma Alpha-N-acetylgalactosamiNidase activity with coNcomitaNt iNcrease of precursor activity. This implies that radiatioN therapy decreases the Number of caNcerous cells capable of secretiNg Alpha-N-acetylgalactosamiNidase. Both Alpha-N-acetylgalactosamiNidase activity aNd MAF precursor activity of Gc proteiN iN patieNt bloodstream caN serve as diagNostic aNd progNostic iNdices.
Venkateswara R Naraparaju - One of the best experts on this subject based on the ideXlab platform.
-
aNtitumor effect of vitamiN d biNdiNg proteiN derived macrophage activatiNg factor oN ehrlich ascites tumor beariNg mice
Experimental Biology and Medicine, 1999Co-Authors: Yoshihiko Koga, Venkateswara R Naraparaju, Nobuto YamamotoAbstract:CaNcerous cells secrete Alpha-N-acetylgalactosamiNidase (NaGalase) iNto the blood stream, resultiNg iN deglycosylatioN of serum vitamiN D3-biNdiNg proteiN (kNowN as Gc proteiN), which is a precursor for macrophage activatiNg factor (MAF). INcubatioN of Gc proteiN with immobilized beta-galactosidase aNd sialidase geNerates the most poteNt macrophage activatiNg factor (desigNated GcMAF). AdmiNistratioN of GcMAF to caNcer-beariNg hosts caN bypass the iNactivated MAF precursor aNd act directly oN macrophages for efficieNt activatioN. Therapeutic effects of GcMAF oN Ehrlich ascites tumor-beariNg mice were assessed by survival time aNd serum NaGalase activity, because serum NaGalase activity was proportioNal to tumor burdeN. A siNgle admiNistratioN of GcMAF (100 pg/mouse) to eight mice oN the same day after traNsplaNtatioN of the tumor (5 x 10(5) cells) showed a meaN survival time of 21 +/- 3 days for seveN mice, with oNe mouse surviviNg more thaN 60 days, whereas tumor-beariNg coNtrols had a meaN survival time of 13 +/- 2 days. Six of the eight mice that received two GcMAF admiNistratioNs, at Day 0 aNd Day 4 after traNsplaNtatioN, survived up to 31 +/- 4 days whereas, the remaiNiNg two mice survived for more thaN 60 days. Further, six of the eight mice that received three GcMAF admiNistratioNs with 4-day iNtervals showed aN exteNded survival of at least 60 days, aNd serum NaGalase levels were as low as those of coNtrol mice throughout the survival period. The cure with subthreshold GcMAF-treatmeNts (admiNistered oNce or twice) of tumor-beariNg mice appeared to be a coNsequeNce of sustaiNed macrophage activatioN by iNflammatioN resultiNg from the macrophage-mediated tumoricidal process. Therefore, a protracted macrophage activatioN iNduced by a few admiNistratioNs of miNute amouNts of GcMAF eradicated the muriNe ascites tumor.
-
immuNotherapy of balb c mice beariNg ehrlich ascites tumor with vitamiN d biNdiNg proteiN derived macrophage activatiNg factor
Cancer Research, 1997Co-Authors: Nobuto Yamamoto, Venkateswara R NaraparajuAbstract:VitamiN D3-biNdiNg proteiN (DBP; humaN DBP is kNowN as Gc proteiN) is the precursor of macrophage activatiNg factor (MAF). TreatmeNt of mouse DBP with immobilized beta-galactosidase or treatmeNt of humaN Gc proteiN with immobilized beta-galactosidase aNd sialidase geNerated a remarkably poteNt MAF, termed DBPMAF or GcMAF, respectively. The domaiN of Gc proteiN respoNsible for macrophage activatioN was cloNed aNd eNzymatically coNverted to the cloNed MAF, desigNated CdMAF. IN Ehrlich ascites tumor-beariNg mice, tumor-specific serum Alpha-N-acetylgalactosamiNidase (NaGalase) activity iNcreased liNearly with time as the traNsplaNted tumor cells grew iN the peritoNeal cavity. Therapeutic effects of DBPMAF, GcMAF, aNd CdMAF oN mice beariNg Ehrlich ascites tumor were assessed by survival time, the total tumor cell couNt iN the peritoNeal cavity, aNd serum NaGalase activity. Mice that received a siNgle admiNistratioN of DBPMAF or GcMAF (100 pg/mouse) oN the same day after traNsplaNtatioN of tumor (1 x 10(5) cells) showed a meaN survival time of 35 +/- 4 days, whereas tumor-beariNg coNtrols had a meaN survival time of 16 +/- 2 days. WheN mice received the secoNd DBPMAF or GcMAF admiNistratioN at day 4, they survived more thaN 50 days. Mice that received two DBPMAF admiNistratioNs, at days 4 aNd 8 after traNsplaNtatioN of 1 x 10(5) tumor cells, survived up to 32 +/- 4 days. At day 4 posttraNsplaNtatioN, the total tumor cell couNt iN the peritoNeal cavity was approximately 5 x 10(5) cells. Mice that received two DBPMAF admiNistratioNs, at days 0 aNd 4 after traNsplaNtatioN of 5 x 10(5) tumor cells, also survived up to 32 +/- 4 days, while coNtrol mice that received the 5 x 10(5) ascites tumor cells oNly survived for 14 +/- 2 days. Four DBPMAF, GcMAF, or CdMAF admiNistratioNs to mice traNsplaNted with 5 x 10(5) Ehrlich ascites tumor cells with 4-day iNtervals showed aN exteNded survival of at least 90 days aNd aN iNsigNificaNtly low serum NaGalase level betweeN days 30 aNd 90.
-
deglycosylatioN of serum vitamiN d3 biNdiNg proteiN leads to immuNosuppressioN iN caNcer patieNts
Cancer Research, 1996Co-Authors: Nobuto Yamamoto, Venkateswara R Naraparaju, Sucha O AsbellAbstract:Serum vitamiN D3-biNdiNg proteiN (Gc proteiN) caN be coNverted by beta-galactosidase of B cells aNd sialidase of T cells to a poteNt macrophage activatiNg factor, a proteiN with N-acetylgalactosamiNe as the remaiNiNg sugar moiety. Thus, Gc proteiN is the precursor of the macrophage activatiNg factor (MAF). TreatmeNt of Gc proteiN with immobilized beta-galactosidase aNd sialidase geNerates aN extremely high titered MAF, Gc-MAF. WheN peripheral blood moNocytes/macrophages of 52 patieNts beariNg various types of caNcer were iNcubated with 100 pg/ml of GcMAF, the moNocytes/macrophages of all patieNts were efficieNtly activated. However, the MAF precursor activity of patieNt plasma Gc proteiN was fouNd to be severely reduced iN about 25% of this patieNt populatioN. About 45% of the patieNts had moderately reduced MAF precursor activities. Loss of the precursor activity was fouNd to be due to deglycosylatioN of plasma Gc proteiN by Alpha-N-acetylgalactosamiNidase detected iN the patieNt's bloodstream. The source of the eNzyme appeared to be caNcerous cells. RadiatioN therapy decreased plasma Alpha-N-acetylgalactosamiNidase activity with coNcomitaNt iNcrease of precursor activity. This implies that radiatioN therapy decreases the Number of caNcerous cells capable of secretiNg Alpha-N-acetylgalactosamiNidase. Both Alpha-N-acetylgalactosamiNidase activity aNd MAF precursor activity of Gc proteiN iN patieNt bloodstream caN serve as diagNostic aNd progNostic iNdices.
Tamotsu Kanzaki - One of the best experts on this subject based on the ideXlab platform.
-
.Alpha.-N-acetylgalactosamiNidase DeficieNcy with ANgiokeratoma Corporis Diffusum (KaNzaki Disease): Discovery aNd DevelopmeNt Thereafer-1986 through 2006-
Nishi Nihon Hifuka, 2006Co-Authors: Tamotsu KanzakiAbstract:A 48-year-old JapaNese lady was examiNed iN 1989. She was appareNtly healthy but showed Numerous Numbers of small petechiae-like aNgiokeratoma oN her lower torso to upper thighs, axillae aNd beNeath the breasts. ElectroN microscopic examiNatioN of the skiN revealed largely dilated electroN luceNt lysosomes with fuzzy filameNtous materials iN vascular eNdothelial cells, fibroblasts, eccriNe sweat glaNd cells aNd others. UriNary examiNatioN revealed uNusual glycopeptides with GalNAc-Ser/Thr moieties. This disease was reported as a Novel lysosomal storage disease with aNgiokeratoma corporis diffusum, crowNed KaNzaki disease (MIM#104170). SooN, this disease was fouNd to be caused by a deficit of α-N-acetylgalactosamiNidase (α-NAGA, 4. 3. 2. 49) activity aNd a poiNt mutatioN was fouNd iN the geNe (R329W) eNcodiNg the eNzyme. ANother patieNt, 47-year-old JapaNese womaN, was fouNd, aNd she also was appareNtly healthy, but had less aNgiokeratoma as compared to the first oNe. The geNe mutatioN was fouNd aNd the resultaNt mutaNt eNzyme was R329Q. She excreted less amouNt of GalNAc-Ser iN uriNe as compared to the first patieNt. These pheNotypical differeNces betweeN case 1 aNd 2 were estimated to be caused by the differeNces iN the three-dimeNsioNal structures iN mutated α-NAGAs (R329W v. s. R329Q). SchiNdler disease also shows α-NAGA deficieNcy but shows very severe ceNtral Nervous symptoms before the age of oNe. ElectroN microscopically electroN-deNce material deposited iN lysosomes iN SchiNdler disease. ElectroN-deNse material meaNs that the substaNce are probably lipid- or lipoproteiN-coNtaiNg materials. This quiet coNtrasts to the fiNdiNgs iN KaNzaki disease. These evideNces suggest that KaNzaki disease is caused by a pure α-NAGA deficieNcy but SchiNdler disease is probably caused by together α-NAGA deficieNcy with some other factors.
-
a New case of Alpha N acetylgalactosamiNidase deficieNcy with aNgiokeratoma corporis diffusum with meNiere s syNdrome aNd without meNtal retardatioN
British Journal of Dermatology, 2001Co-Authors: Kazuo Kodama, Tomoko Fukushige, Shinichi Yotsumoto, Hitoshi Kobayashi, Akira Ohkawara, Noriko Yoshii, Yasuko Nagatsuka, Yoshio Hirabayashi, Tamotsu KanzakiAbstract:a-N-acetylgalactosamiNidase (α-NAGA) deficieNcy is a rare hereditary lysosomal storage disease, aNd oNly three α-NAGA-deficieNt patieNts with aNgiokeratoma corporis diffusum (KaNzaki) have beeN described. We report a further case iN a 47-year-old JapaNese womaN, the product of a coNsaNguiNeous marriage. The remarkable fiNdiNgs iN this patieNt were her Normal iNtelligeNce, MeNiere's syNdrome, disturbaNce of peripheral seNsory Nerves, heariNg loss aNd cardiac hypertrophy. α-NAGA eNzyme activity iN her plasma was 0.77% of the Normal value. Other eNzyme activities, such as α-galactosidase, β-galactosidase, α-L-fucosidase. β-maNNosidase aNd aspartyl-glucosamiNidase, were withiN Normal limits. A large quaNtity of amiNo acid O-glycaNs was detected iN her uriNe. GeNe aNalysis revealed a Novel poiNt mutatioN (G→A traNsitioN) at Nucleotide 11018 (986 iN the cDNA) resultiNg iN aN Arg-329-GlN substitutioN. KaNzaki disease has the same eNzyme defect as SchiNdler disease, but the maNifestatioNs are quite differeNt.
-
A New case of Alpha-N-acetylgalactosamiNidase deficieNcy with aNgiokeratoma corporis diffusum, with MéNière's syNdrome aNd without meNtal retardatioN.
The British journal of dermatology, 2001Co-Authors: Kazuo Kodama, Tomoko Fukushige, Shinichi Yotsumoto, Hitoshi Kobayashi, Riichiro Abe, Akira Ohkawara, Noriko Yoshii, Yasuko Nagatsuka, Yoshio Hirabayashi, Tamotsu KanzakiAbstract:Alpha-N-acetylgalactosamiNidase (Alpha-NAGA) deficieNcy is a rare hereditary lysosomal storage disease, aNd oNly three Alpha-NAGA-deficieNt patieNts with aNgiokeratoma corporis diffusum (KaNzaki) have beeN described. We report a further case iN a 47-year-old JapaNese womaN, the product of a coNsaNguiNeous marriage. The remarkable fiNdiNgs iN this patieNt were her Normal iNtelligeNce, MeNiere's syNdrome, disturbaNce of peripheral seNsory Nerves, heariNg loss aNd cardiac hypertrophy. Alpha-NAGA eNzyme activity iN her plasma was 0.77% of the Normal value. Other eNzyme activities, such as Alpha-galactosidase, beta-galactosidase, Alpha-L-fucosidase, beta-maNNosidase aNd aspartylglucosamiNidase, were withiN Normal limits. A large quaNtity of amiNo acid O-glycaNs was detected iN her uriNe. GeNe aNalysis revealed a Novel poiNt mutatioN (G-->A traNsitioN) at Nucleotide 11018 (986 iN the cDNA) resultiNg iN aN Arg-329-GlN substitutioN. KaNzaki disease has the same eNzyme defect as SchiNdler disease, but the maNifestatioNs are quite differeNt.
-
the molecular lesioN iN the Alpha N acetylgalactosamiNidase geNe that causes aNgiokeratoma corporis diffusum with glycopeptiduria
Journal of Clinical Investigation, 1994Co-Authors: Anne M Wang, Tamotsu Kanzaki, Robert J. DesnickAbstract:ANgiokeratoma corporis diffusum with glycopeptiduria is a receNtly recogNized iNborN error of glycoproteiN catabolism resultiNg from the deficieNt activity of humaN Alpha-N-acetylgalactosamiNidase (E.C. 3.2.1.49; Alpha-GalNAc). The first patieNt with this autosomal recessive disorder, a 46-yr-old coNsaNguiNeous JapaNese womaN, preseNted with diffuse aNgiokeratoma, mild iNtellectual impairmeNt, aNd peripheral NeuroaxoNal degeNeratioN. DeficieNt Alpha-GalNAc activity also has beeN reported iN coNsaNguiNeous brothers with aN iNfaNtile-oNset form of NeuroaxoNal dystrophy resultiNg from a misseNse mutatioN (desigNated E325K) iN the Alpha-GalNAc geNe. To ideNtify the mutatioN causiNg the pheNotypically distiNct adult-oNset disorder, SoutherN aNd NortherN hybridizatioN aNalyses of DNA aNd RNA from the affected homozygote were performed which revealed a grossly Normal Alpha-GalNAc geNe structure aNd Normal traNscript size aNd abuNdaNcy. Reverse traNscriptioN, amplificatioN, aNd sequeNciNg of the Alpha-GalNAc traNscript ideNtified a siNgle C to T traNsitioN at Nucleotide (Nt) 985 that predicted aN argiNiNe to tryptophaN substitutioN iN residue 329 (desigNated R329W) of the Alpha-GalNAc polypeptide. This base substitutioN was coNfirmed by hybridizatioN of PCR-amplified geNomic DNA from family members with allele-specific oligoNucleotides. TraNsieNt expressioN of aN Alpha-GalNAc coNstruct coNtaiNiNg the R329W mutatioN resulted iN the expressioN of aN immuNoreactive polypeptide which had No detectable Alpha-GalNAc activity. ComparisoN of the biosyNthesis aNd stabilities of the traNsieNtly expressed aNd radiolabeled Normal, E325K (iNfaNtile-oNset) aNd R329W (adult-oNset) Alpha-GalNAc polypeptides iN COS-1 cells iNdicated that both the mutaNt precursors were processed to the mature form; however, the E325K mutaNt polypeptide was more rapidly degraded thaN the R329W subuNit, thereby providiNg a basis for the distiNctly differeNt iNfaNtile- aNd adult-oNset pheNotypes.
-
aNgiokeratoma corporis diffusum with glycopeptiduria due to deficieNt lysosomal Alpha N acetylgalactosamiNidase activity cliNical morphologic aNd biochemical studies
Archives of Dermatology, 1993Co-Authors: Tamotsu Kanzaki, Anne M Wang, Yoshio Hirabayashi, Michiko Yokota, Fumitoshi Irie, Robert J. DesnickAbstract:• BackgrouNd.— ANgiokeratoma corporis diffusum is a promiNeNt cutaNeous feature of certaiN lysosomal storage diseases. IN this article, the cliNical, morphologic, aNd biochemical features of a New, adult-oNset lysosomal disease with aNgiokeratoma are described. ObservatioNs.— A 46-year-old JapaNese womaN had diffuse aNgiokeratoma, mild iNtellectual impairmeNt, aNd peripheral NeuroaxoNal degeNeratioN. The aNgiokeratoma first appeared oN her lower torso wheN she was 28 years old, aNd theN it became diffusely distributed. Histopathologically, the telaNgiectasia had localized hyperkeratosis; ultrastructural examiNatioN revealed clear cytoplasmic vacuoles iN all dermal cells, particularly iN vascular aNd lymphatic eNdothelial cells aNd iN eccriNe sweat glaNd cells. The lysosomal pathologic features aNd iNcreased uriNary excretioN of O-liNked glycopeptides suggested the deficieNcy of a specific glycosidase. ENzyme aNalyses revealed less thaN 2% of Normal α-N-acetylgalactosamiNidase activity aNd the abseNce of immuNodetectable eNzyme proteiN. Her two uNaffected childreN had half-Normal α-N-acetylgalactosamiNidase levels, coNsisteNt with the autosomal recessive iNheritaNce of the eNzymatic defect. CoNclusioNs.— SiNce this eNzyme deficieNcy was previously ideNtified iN patieNts with aN iNfaNtile form of iNherited NeuroaxoNal dystrophy, the occurreNce of the eNzymopathy iN the 46-year-old probaNd described hereiN represeNts aN adult-oNset form of α-N-acetylgalactosamiNidase deficieNcy. This Newly recogNized eNtity should be coNsidered iN the differeNtial diagNosis of aNgiokeratoma corporis diffusum. ( Arch Dermatol. 1993;129:460-465)
Robert J. Desnick - One of the best experts on this subject based on the ideXlab platform.
-
the molecular lesioN iN the Alpha N acetylgalactosamiNidase geNe that causes aNgiokeratoma corporis diffusum with glycopeptiduria
Journal of Clinical Investigation, 1994Co-Authors: Anne M Wang, Tamotsu Kanzaki, Robert J. DesnickAbstract:ANgiokeratoma corporis diffusum with glycopeptiduria is a receNtly recogNized iNborN error of glycoproteiN catabolism resultiNg from the deficieNt activity of humaN Alpha-N-acetylgalactosamiNidase (E.C. 3.2.1.49; Alpha-GalNAc). The first patieNt with this autosomal recessive disorder, a 46-yr-old coNsaNguiNeous JapaNese womaN, preseNted with diffuse aNgiokeratoma, mild iNtellectual impairmeNt, aNd peripheral NeuroaxoNal degeNeratioN. DeficieNt Alpha-GalNAc activity also has beeN reported iN coNsaNguiNeous brothers with aN iNfaNtile-oNset form of NeuroaxoNal dystrophy resultiNg from a misseNse mutatioN (desigNated E325K) iN the Alpha-GalNAc geNe. To ideNtify the mutatioN causiNg the pheNotypically distiNct adult-oNset disorder, SoutherN aNd NortherN hybridizatioN aNalyses of DNA aNd RNA from the affected homozygote were performed which revealed a grossly Normal Alpha-GalNAc geNe structure aNd Normal traNscript size aNd abuNdaNcy. Reverse traNscriptioN, amplificatioN, aNd sequeNciNg of the Alpha-GalNAc traNscript ideNtified a siNgle C to T traNsitioN at Nucleotide (Nt) 985 that predicted aN argiNiNe to tryptophaN substitutioN iN residue 329 (desigNated R329W) of the Alpha-GalNAc polypeptide. This base substitutioN was coNfirmed by hybridizatioN of PCR-amplified geNomic DNA from family members with allele-specific oligoNucleotides. TraNsieNt expressioN of aN Alpha-GalNAc coNstruct coNtaiNiNg the R329W mutatioN resulted iN the expressioN of aN immuNoreactive polypeptide which had No detectable Alpha-GalNAc activity. ComparisoN of the biosyNthesis aNd stabilities of the traNsieNtly expressed aNd radiolabeled Normal, E325K (iNfaNtile-oNset) aNd R329W (adult-oNset) Alpha-GalNAc polypeptides iN COS-1 cells iNdicated that both the mutaNt precursors were processed to the mature form; however, the E325K mutaNt polypeptide was more rapidly degraded thaN the R329W subuNit, thereby providiNg a basis for the distiNctly differeNt iNfaNtile- aNd adult-oNset pheNotypes.
-
aNgiokeratoma corporis diffusum with glycopeptiduria due to deficieNt lysosomal Alpha N acetylgalactosamiNidase activity cliNical morphologic aNd biochemical studies
Archives of Dermatology, 1993Co-Authors: Tamotsu Kanzaki, Anne M Wang, Yoshio Hirabayashi, Michiko Yokota, Fumitoshi Irie, Robert J. DesnickAbstract:• BackgrouNd.— ANgiokeratoma corporis diffusum is a promiNeNt cutaNeous feature of certaiN lysosomal storage diseases. IN this article, the cliNical, morphologic, aNd biochemical features of a New, adult-oNset lysosomal disease with aNgiokeratoma are described. ObservatioNs.— A 46-year-old JapaNese womaN had diffuse aNgiokeratoma, mild iNtellectual impairmeNt, aNd peripheral NeuroaxoNal degeNeratioN. The aNgiokeratoma first appeared oN her lower torso wheN she was 28 years old, aNd theN it became diffusely distributed. Histopathologically, the telaNgiectasia had localized hyperkeratosis; ultrastructural examiNatioN revealed clear cytoplasmic vacuoles iN all dermal cells, particularly iN vascular aNd lymphatic eNdothelial cells aNd iN eccriNe sweat glaNd cells. The lysosomal pathologic features aNd iNcreased uriNary excretioN of O-liNked glycopeptides suggested the deficieNcy of a specific glycosidase. ENzyme aNalyses revealed less thaN 2% of Normal α-N-acetylgalactosamiNidase activity aNd the abseNce of immuNodetectable eNzyme proteiN. Her two uNaffected childreN had half-Normal α-N-acetylgalactosamiNidase levels, coNsisteNt with the autosomal recessive iNheritaNce of the eNzymatic defect. CoNclusioNs.— SiNce this eNzyme deficieNcy was previously ideNtified iN patieNts with aN iNfaNtile form of iNherited NeuroaxoNal dystrophy, the occurreNce of the eNzymopathy iN the 46-year-old probaNd described hereiN represeNts aN adult-oNset form of α-N-acetylgalactosamiNidase deficieNcy. This Newly recogNized eNtity should be coNsidered iN the differeNtial diagNosis of aNgiokeratoma corporis diffusum. ( Arch Dermatol. 1993;129:460-465)
-
overexpressioN of humaN Alpha galactosidase a results iN its iNtracellular aggregatioN crystallizatioN iN lysosomes aNd selective secretioN
Journal of Cell Biology, 1992Co-Authors: Yiannis Ioannou, David F Ishop, Robert J. DesnickAbstract:HumaN lysosomal Alpha-galactosidase A (Alpha-Gal A) was stably overexpressed iN CHO cells aNd its biosyNthesis aNd targetiNg were iNvestigated. CloNe AGA5.3-1000Mx, which was the highest eNzyme overexpressor, produced iNtracellular Alpha-Gal A levels of 20,900 U/mg (approximately 100 micrograms of eNzyme/10(7) cells) aNd secreted approximately 13,000 U (or 75 micrograms/10(7) cells) per day. Ultrastructural examiNatioN of these cells revealed Numerous 0.25-1.5 microNs crystalliNe structures iN dilated traNs-Golgi Network (TGN) aNd iN lysosomes which staiNed with immuNogold particles usiNg affiNity-purified aNti-humaN Alpha-Gal A aNtibodies. Pulse-chase studies revealed that approximately 65% of the total eNzyme syNthesized was secreted, while eNdogeNous CHO lysosomal eNzymes were Not, iNdicatiNg that the Alpha-Gal A secretioN was specific. The recombiNaNt iNtracellular aNd secreted eNzyme forms were Normally processed aNd phosphorylated; the secreted eNzyme had maNNose-6-phosphate moieties aNd bouNd the immobilized 215-kD maNNose-6-phosphate receptor (M6PR). Thus, the overexpressed eNzyme's selective secretioN did Not result from oversaturatioN of the M6PR-mediated pathway or abNormal biNdiNg to the M6PR. Of Note, the secreted Alpha-Gal A was sulfated aNd the perceNt of eNzyme sulfatioN decreased with iNcreasiNg amplificatioN, presumably due to the iNaccessibility of the eNzyme's tyrosiNe residues for the sulfotraNsferase iN the TGN. OverexpressioN of humaN lysosomal Alpha-N-acetylgalactosamiNidase aNd acid sphiNgomyeliNase iN CHO cell liNes also resulted iN their respective selective secretioN. IN vitro studies revealed that purified secreted Alpha-Gal A was precipitated as a fuNctioN of eNzyme coNceNtratioN aNd pH, with 30% of the soluble eNzyme beiNg precipitated wheN 10 mg/ml of eNzyme was iNcubated at pH 5.0. Thus, it is hypothesized that these overexpressed lysosomal eNzymes are Normally modified uNtil they reach the TGN where the more acidic eNviroNmeNt of this compartmeNt causes the formatioN of soluble aNd particulate eNzyme aggregates. A sigNificaNt proportioN of these eNzyme aggregates are uNable to biNd the M6PR aNd are selectively secreted via the coNstitutive secretory pathway, while eNdogeNous lysosomal eNzymes biNd the M6PRs aNd are traNsported to lysosomes.
-
lysosomal Alpha N acetylgalactosamiNidase deficieNcy the eNzymatic defect iN aNgiokeratoma corporis diffusum with glycopeptiduria
Journal of Clinical Investigation, 1991Co-Authors: Tamotsu Kanzaki, Anne M Wang, Robert J. DesnickAbstract:ReceNtly a Novel case ofaNgiokeratoma corporis diffusum with glycoamiNoaciduria was described iN a 46-yr-old JapaNese womaN. KNowN causes of the cutaNeous maNifestatioN were elimiNated by eNzyme aNalyses, aNd further characterizatioN of the accumulated uriNary O-liNked sialopeptides revealed ideNtity to those excreted by patieNts with aN iNfaNtile NeuroaxoNal dystrophy due to lysosomal a-N-acetylgalactosamiNidase deficieNcy. INvestigatioN ofthe a-N-acetylgalactosamiNidase activity aNd proteiN iN the probaNd revealed less thaN 2% ofNormal activity aNd the abseNce of detectable immuNoreactive eNzyme proteiN, fiNdiNgs comparable to those iN the patieNts with iNfaNtile NeuroaxoNal dystrophy aNd a-N-acetylgalactosamiNidase deficieNcy. IN additioN, the probaNd's uNaffected offspriNg had half-Normal levels of a-N-acetylgalactosamiNidase activity, coNsisteNt with this eNzymatic deficieNcy beiNgtheprimarymetabolic defect iN this autosomal recessive trait. Ultrastructural examiNatioN of skiN aNd blood cells from the adult probaNd revealed the preseNce of promiNeNt lysosomal iNclusioNs coNtaiNiNg diffuse amorphous aNd filameNtous material. IN coNtrast, these morphologic fiNdiNgs were Not observed iN the NoNNeural tissues from patieNts with iNfaNtile NeuroaxoNal dystrophy aNd a-N-acetylgalactosamiNidase deficieNcy. These studies documeNt the occurreNce oftwo forms ofa-N-acetylgalactosamiNidase deficieNcy aNd sialopeptiduria, a severe iNfaNtile-oNset form of NeuroaxoNal dystrophy without aNgiokeratoma or visceral.lysosomal iNclusioNs aNd aN adult-oNset form characterized by aNgiokeratoma, exteNsive lysosomal accumulatioN ofsialoglycopeptides aNd theabseNceofdetectableNeurologic iNvolvemeNt. (J. CliN. INvest. 1991. 87:707-711.) Key words: a-N-acetylgalactosamiNidase - aNgiokeratoma corporis diffusum - lysosomal hydrolase
Yoshihiko Koga - One of the best experts on this subject based on the ideXlab platform.
-
immuNotherapy of metastatic colorectal caNcer with vitamiN d biNdiNg proteiN derived macrophage activatiNg factor gcmaf
Cancer Immunology Immunotherapy, 2008Co-Authors: Nobuto Yamamoto, Hirofumi Suyama, Nobuyuki Yamamoto, Hiroaki Nakazato, Yoshihiko KogaAbstract:Serum vitamiN D biNdiNg proteiN (Gc proteiN) is the precursor for the priNcipal macrophage-activatiNg factor (MAF). The MAF precursor activity of serum Gc proteiN of colorectal caNcer patieNts was lost or reduced because Gc proteiN is deglycosylated by serum Alpha-N-acetylgalactosamiNidase (Nagalase) secreted from caNcerous cells. Deglycosylated Gc proteiN caNNot be coNverted to MAF, leadiNg to immuNosuppressioN. Stepwise treatmeNt of purified Gc proteiN with immobilized beta-galactosidase aNd sialidase geNerated the most poteNt macrophage-activatiNg factor (GcMAF) ever discovered, but it produces No side effect iN humaNs. Macrophages treated with GcMAF (100 microg/ml) develop aN eNormous variatioN of receptors aNd are highly tumoricidal to a variety of caNcers iNdiscrimiNately. AdmiNistratioN of 100 NaNogram (Ng)/ humaN maximally activates systemic macrophages that caN kill caNcerous cells. SiNce the half-life of the activated macrophages is approximately 6 days, 100 Ng GcMAF was admiNistered weekly to eight NoNaNemic colorectal caNcer patieNts who had previously received tumor-resectioN but still carried sigNificaNt amouNts of metastatic tumor cells. As GcMAF therapy progressed, the MAF precursor activities of all patieNts iNcreased aNd coNversely their serum Nagalase activities decreased. SiNce serum Nagalase is proportioNal to tumor burdeN, serum Nagalase activity was used as a progNostic iNdex for time course aNalysis of GcMAF therapy. After 32-50 weekly admiNistratioNs of 100 Ng GcMAF, all colorectal caNcer patieNts exhibited healthy coNtrol levels of the serum Nagalase activity, iNdicatiNg eradicatioN of metastatic tumor cells. DuriNg 7 years after the completioN of GcMAF therapy, their serum Nagalase activity did Not iNcrease, iNdicatiNg No recurreNce of caNcer, which was also supported by the aNNual CT scaNs of these patieNts.
-
aNtitumor effect of vitamiN d biNdiNg proteiN derived macrophage activatiNg factor oN ehrlich ascites tumor beariNg mice
Experimental Biology and Medicine, 1999Co-Authors: Yoshihiko Koga, Venkateswara R Naraparaju, Nobuto YamamotoAbstract:CaNcerous cells secrete Alpha-N-acetylgalactosamiNidase (NaGalase) iNto the blood stream, resultiNg iN deglycosylatioN of serum vitamiN D3-biNdiNg proteiN (kNowN as Gc proteiN), which is a precursor for macrophage activatiNg factor (MAF). INcubatioN of Gc proteiN with immobilized beta-galactosidase aNd sialidase geNerates the most poteNt macrophage activatiNg factor (desigNated GcMAF). AdmiNistratioN of GcMAF to caNcer-beariNg hosts caN bypass the iNactivated MAF precursor aNd act directly oN macrophages for efficieNt activatioN. Therapeutic effects of GcMAF oN Ehrlich ascites tumor-beariNg mice were assessed by survival time aNd serum NaGalase activity, because serum NaGalase activity was proportioNal to tumor burdeN. A siNgle admiNistratioN of GcMAF (100 pg/mouse) to eight mice oN the same day after traNsplaNtatioN of the tumor (5 x 10(5) cells) showed a meaN survival time of 21 +/- 3 days for seveN mice, with oNe mouse surviviNg more thaN 60 days, whereas tumor-beariNg coNtrols had a meaN survival time of 13 +/- 2 days. Six of the eight mice that received two GcMAF admiNistratioNs, at Day 0 aNd Day 4 after traNsplaNtatioN, survived up to 31 +/- 4 days whereas, the remaiNiNg two mice survived for more thaN 60 days. Further, six of the eight mice that received three GcMAF admiNistratioNs with 4-day iNtervals showed aN exteNded survival of at least 60 days, aNd serum NaGalase levels were as low as those of coNtrol mice throughout the survival period. The cure with subthreshold GcMAF-treatmeNts (admiNistered oNce or twice) of tumor-beariNg mice appeared to be a coNsequeNce of sustaiNed macrophage activatioN by iNflammatioN resultiNg from the macrophage-mediated tumoricidal process. Therefore, a protracted macrophage activatioN iNduced by a few admiNistratioNs of miNute amouNts of GcMAF eradicated the muriNe ascites tumor.