ABSTRACT
A 79-year-old woman was referred to our hospital because numerous polyps were found in her stomach and large intestine at an ambulatory clinic. Although there were no characteristic symptoms or signs of Cronkhite-Canada syndrome (CCS), endoscopic and pathological findings indicated CCS. Moreover, colonoscopy showed two polypoid lesions (Is type), which appeared neoplastic by magnifying observation with image-enhanced endoscopy (IEE), in the ascending colon. Histologically, the resected specimens revealed tubular adenomas arising in the CCS inflammatory polyps. Remarkable remission of the polyps and edematous mucosa in the stomach and colon was seen after 8 months of administration of salazosulfapyridine (SASP) (3 g/day). Another adenoma was detected and removed endoscopically in the sigmoid colon. This is the first report to describe an asymptomatic case of CCS probably detected in the early phase of the disease, by magnifying IEE which enabled detection and treatment for associated colonic adenomas. SASP was effective in eradication of the inflammatory polyposis, and an additional adenoma was successfully found and removed by surveillance colonoscopy thereafter.
Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/therapeutic use , Endoscopy, Gastrointestinal/methods , Intestinal Polyposis/therapy , Mucous Membrane/pathology , Sulfasalazine/therapeutic use , Aged , Combined Modality Therapy , Female , Humans , Intestinal Polyposis/drug therapy , Intestinal Polyposis/surgery , Mucous Membrane/surgery , Treatment OutcomeABSTRACT
A 74-year-old woman underwent colonoscopy for investigation of a liver tumor. A lateral spreading tumor of the non-granular type (LST-NG), 25 mm in diameter, was detected at the rectosigmoid junction. As magnifying image-enhanced colonoscopy suggested a tubulovillous adenoma, endoscopic mucosal resection (EMR) was chosen for removal of the LST-NG. The lesion was effectively and evenly lifted after injection of 0.4% hyaluronic acid diluted with glycerol in the ratio of 1:1. A small amount of indigo-carmine dye was also added for coloration of the plane of resection. The lesion was completely removed en bloc. Although a blue-colored layer was identified in the resection defect, a small amount of a whitish layer was detected above the blue layer. The muscle layer was clearly located on the underside of the resected polyp. A total of 14 endoclips were used to close the defect completely. The patient was successfully treated conservatively without surgery. Histology of the resected specimen showed that it contained a tubulovillous adenoma with the submucosal layer and both layers of the muscularis propria. The surgical margin was free of neoplastic change horizontally and vertically. To the best of our knowledge, this is the first case report of full-thickness resection associated with EMR after unplanned injection of dilute hyaluronic acid into the subserosal layer rather than the intended submucosal layer. We describe how to promptly recognize this complication during colonoscopy, in order to achieve immediate closure of the defect, with the identification of a "mirror target sign" on the colonic wall.
Subject(s)
Adenoma, Villous/surgery , Hyaluronic Acid/administration & dosage , Intestinal Mucosa/surgery , Medical Errors , Rectal Neoplasms/surgery , Adenoma, Villous/pathology , Aged , Colonoscopy , Female , Humans , Intestinal Mucosa/pathology , Rectal Neoplasms/pathologyABSTRACT
AIMS: The use of vitamin E-infused highly crosslinked polyethylene (HXLPE) in total knee prostheses is controversial. In this paper we have compared the clinical and radiological results between conventional polyethylene and vitamin E-infused HXLPE inserts in total knee arthroplasty (TKA). PATIENTS AND METHODS: The study included 200 knees (175 patients) that underwent TKA using the same total knee prostheses. In all, 100 knees (77 patients) had a vitamin E-infused HXLPE insert (study group) and 100 knees (98 patients) had a conventional polyethylene insert (control group). There were no significant differences in age, sex, diagnosis, preoperative knee range of movement (ROM), and preoperative Knee Society Score (KSS) between the two groups. Clinical and radiological results were evaluated at two years postoperatively. RESULTS: Differences in postoperative ROM and KSS were not statistically significant between the study and control groups. No knee exhibited osteolysis, aseptic loosening, or polyethylene failure. Additionally, there was no significant difference in the incidence of a radiolucent line between the two groups. One patient from the study group required irrigation and debridement, due to deep infection, at six months postoperatively. CONCLUSION: Clinical results were comparable between vitamin E-infused HXLPE inserts and conventional polyethylene inserts at two years after TKA, without any significant clinical failure. Cite this article: Bone Joint J 2019;101-B:559-564.
Subject(s)
Arthroplasty, Replacement, Knee/instrumentation , Knee Prosthesis/adverse effects , Polyethylene/administration & dosage , Prosthesis Design/methods , Vitamin E/administration & dosage , Aged , Arthroplasty, Replacement, Knee/adverse effects , Arthroplasty, Replacement, Knee/methods , Cross-Linking Reagents , Female , Humans , Knee Joint/surgery , Male , Middle Aged , Postoperative Period , Prosthesis Design/adverse effects , Range of Motion, ArticularSubject(s)
Adenoma/surgery , Appendiceal Neoplasms/surgery , Appendix/surgery , Colonoscopes , Colonoscopy/methods , Dissection/instrumentation , Intestinal Mucosa/surgery , Adenoma/diagnosis , Appendiceal Neoplasms/diagnosis , Appendix/pathology , Diagnosis, Differential , Equipment Design , Humans , Intestinal Mucosa/pathology , Male , Middle AgedABSTRACT
We describe a 39-year-old Japanese man with rippling muscle disease who carried a novel homozygous mutation (Trp70 to a stop codon) in the caveolin-3 gene. The patient also had extraocular muscle paresis showing atrophy of the extraocular muscles on orbital MRI. The involvement of the extraocular muscles of patients with caveolinopathy is discussed.
Subject(s)
Caveolin 3/genetics , Homozygote , Muscular Diseases/genetics , Mutation/genetics , Ocular Motility Disorders/genetics , Adult , Codon, Terminator/genetics , DNA Mutational Analysis , Humans , Magnetic Resonance Imaging , Male , Muscular Diseases/pathology , Ocular Motility Disorders/pathology , Oculomotor Muscles/pathology , Oculomotor Muscles/physiopathology , Tryptophan/geneticsABSTRACT
A recombinant phage containing an actin gene (lambda Ha201) was isolated from a human DNA library and the structure of the actin gene was determined. The amino acid sequences deduced from the nucleotide sequences of lambda Ha201 were compared with those of six actin isoforms; they matched those of bovine aortic smooth muscle actin, except for codon 309, which was valine (GTC) in lambda Ha201 and alanine (GCN) in bovine aortic smooth muscle actin. Southern blot hybridization experiments showed that the gene of normal human cells did not have the TaqI-sensitive site around position 309, whereas half of the genes of HUT14 cells did. These results indicate that one allele of the aortic smooth muscle actin gene in HUT14 cells has a transition point mutation (C----T) at codon 309 and that the amino acid sequences of normal human aorta and bovine smooth muscle actins are probably identical. In addition to the five introns interrupting exons at codons 150, 204, and 267, and between codons 41 and 42 and 327 and 328, which are common to skeletal muscle and cardiac muscle actin genes, the smooth muscle actin gene has two more intron sites between codons 84 and 85 and 121 and 122. The previously unreported intron site between codons 84 and 85 is unique to the smooth muscle actin gene. The intron site between codons 121 and 122 is common to beta-actin genes but is not found in other muscle actin genes. A hypothesis is proposed for the evolutionary pathway of the actin gene family.
Subject(s)
Actins/genetics , Genes , Muscle, Smooth, Vascular/metabolism , Amino Acid Sequence , Aorta/metabolism , Base Sequence , Cloning, Molecular , DNA/metabolism , DNA Restriction Enzymes , DNA, Recombinant/metabolism , Humans , Nucleic Acid Hybridization , Plasmids , RNA, Messenger/geneticsABSTRACT
Recombinant phages that carry the human smooth muscle (enteric type) gamma-actin gene were isolated from human genomic DNA libraries. The amino acid sequence deduced from the nucleotide sequence matches those of cDNAs but differs from the protein sequence previously reported at one amino acid position, codon 359. The gene containing one 5' untranslated exon and eight coding exons extends for 27 kb on human chromosome 2. The intron between codons 84 and 85 (site 3) is unique to the two smooth muscle actin genes. In the 5' flanking region, there are several CArG boxes and E boxes, which are regulatory elements in some muscle-specific genes. Hybridization with the 3' untranslated region, which is specific for the human smooth muscle gamma-actin gene, suggests the single gene in the human genome and specific expressions in enteric and aortic tissues. From characterized molecular structures of the six human actin isoform genes, we propose a hypothesis of evolutionary pathway of the actin gene family. A presumed ancestral actin gene had introns at least sites 1, 2, and 4 through 8. Cytoplasmic actin genes may have directly evolved from it through loss of introns at sites 5 and 6. However, through duplication of the ancestral actin gene with substitutions of many amino acids, a prototype of muscle actin genes had been created. Subsequently, striated muscle actin and smooth muscle actin genes may have evolved from this prototype by loss of an intron at site 4 and acquisition of a new intron at site 3, respectively.
Subject(s)
Actins/genetics , Biological Evolution , Chromosomes, Human, Pair 2 , Muscle, Smooth/physiology , Amino Acid Sequence , Animals , Base Sequence , Blotting, Northern , Blotting, Southern , Cell Line , Chromosome Mapping , DNA/genetics , DNA/isolation & purification , Digestive System Physiological Phenomena , Exons , Genomic Library , Humans , Hybrid Cells/cytology , Hybrid Cells/physiology , Introns , Mice , Molecular Sequence Data , RNA/genetics , RNA/isolation & purification , Rats , Sequence Homology, Nucleic AcidABSTRACT
Cancer patients with weight loss showed urinary excretion of a lipid-mobilizing factor (LMF), determined by the ability to stimulate lipolysis in isolated murine epididymal adipocytes. Such bioactivity was not detectable in the urine of cancer patients without weight loss or in normal subjects. The LMF was purified using a combination of ion exchange, exclusion, and hydrophobic interaction chromatographies to give a single component of apparent Mr 43,000, which showed homology in amino acid sequence with human plasma Zn-alpha2-glycoprotein. Both substances showed the same mobility on denaturing and nondenaturing gels and the same chymotrypsin digestion pattern, both stained heavily for carbohydrate, and they showed similar immunoreactivity. Polyclonal antisera to human plasma Zn-alpha2-glycoprotein was also capable of neutralization of the bioactivity of human LMF in vitro. Using competitive PCR to quantify expression of Zn-alpha2-glycoprotein, we found that only those tumors that were capable of producing a decrease in carcass lipid expressed mRNA for Zn-alpha2-glycoprotein. These results provide strong evidence to suggest that tumor production of Zn-alpha2-glycoprotein is responsible for the lipid catabolism seen in cancer patients.
Subject(s)
Cachexia/urine , Digestive System Neoplasms/urine , Glycoproteins/urine , Lipid Mobilization , Ovarian Neoplasms/urine , Peptides/urine , Seminal Plasma Proteins , Adipocytes/drug effects , Animals , Antibodies, Monoclonal/metabolism , Blood Proteins/chemistry , Blood Proteins/metabolism , Cachexia/complications , Cells, Cultured , Chromatography, Ion Exchange , Digestive System Neoplasms/complications , Epididymis , Female , Glycoproteins/genetics , Humans , Lipolysis/drug effects , Male , Mice , Mice, Inbred Strains , Molecular Weight , Neoplasms, Experimental/urine , Ovarian Neoplasms/complications , Peptides/isolation & purification , Peptides/pharmacology , Polymerase Chain Reaction , Proteoglycans , Zn-Alpha-2-GlycoproteinABSTRACT
This study was designed to clarify the role of endogenous Bcl-xL expression in modulating apoptosis of malignant cells. Administration of bcl-x-antisense oligonucleotides decreased Bcl-xL protein levels in the MKN-45 human gastric cancer cell line. The decrease in Bcl-xL protein content resulted in increased cell death induced by serum deprivation or Fas-antibody administration. Flow cytometric analysis revealed that the increased apoptotic cell death was more prominent in bcl-x-antisense-treated cells as compared to control cells, bcl-x-sense-treated cells, or bcl-x-nonsense-treated cells. To inhibit the effect of intrinsic Bcl-xL protein, we overexpressed Bak, which binds Bcl-xL and inhibits the anti-apoptotic effect of Bcl-xL, by transfection into MKN-45 cells. Bak-overexpressing cells showed increased apoptotic cell death induced by Fas-antibody when compared to parent cells and MKN-neo-transfected cells. Bak-overexpressing cells also showed greater sensitization to 5-fluorouracil and cisplatin than parent cells and MKN-neo-transfected cells. In conclusion, we demonstrated that administration of bcl-x-antisense oligonucleotides or overexpression of Bak protein induces sensitization to apoptosis in MKN-45 gastric cancer cells, suggesting that endogenous Bcl-xL expression in cancer cells is an important modulator of apoptosis.
Subject(s)
Apoptosis/genetics , Gene Expression Regulation, Neoplastic , Neoplasm Proteins/physiology , Proto-Oncogene Proteins c-bcl-2/physiology , Stomach Neoplasms/pathology , Antibodies, Monoclonal/pharmacology , Antineoplastic Agents/pharmacology , Cisplatin/pharmacology , Culture Media, Serum-Free , Fluorouracil/pharmacology , Humans , Membrane Proteins/genetics , Membrane Proteins/physiology , Oligonucleotides, Antisense/pharmacology , Proto-Oncogene Proteins c-bcl-2/genetics , Recombinant Fusion Proteins/physiology , Stomach Neoplasms/genetics , Stomach Neoplasms/metabolism , Tumor Cells, Cultured/drug effects , bcl-2 Homologous Antagonist-Killer Protein , bcl-X Protein , fas Receptor/immunology , fas Receptor/physiologyABSTRACT
BACKGROUND-OBJECTIVE: No study has focused on the difference in efficacy of maintenance therapy between patients with new-onset and recurrent gastroesophageal reflux disease (GERD). The aim of this study is to reveal this point. METHODS: Endoscopically proven GERD patients who had completed 8-week initial therapy were sequentially randomized to continuous arm (Omeprazole 20mg od) or on-demand arm (Omeprazole 20mg on-demand). Patients filled in daily symptoms and tablet usages for 24 weeks. Patients underwent upper GI endoscopy at 24 weeks. Symptom relief was defined as no symptoms for>6 days during a week. The numbers of patients who achieved symptom relief and mucosal healing were compared between the new-onset and recurrent groups in the continuous arm and in the on-demand arm, respectively. RESULTS: Among new-onset GERD [n=82 (continuous: 42 patients, on-demand: 40)], continuous arm achieved significant symptom-relief than in on-demand arm at 4*,5*,6** and 17*week. Among recurrent GERD [n=36(continuous: 17 patients, on-demand: 19)], continuous arm achieved significant symptom-relief at 1**,2*,3*,4*,5**,7**,8**,17* and 18* week, respectively (*<0.05,**<0.01). The number of healed patients was significantly higher in new-onset group (60/68, 88.2%) than in recurrent group (17/30, 56.7%) (<0.01). CONCLUSION: Since therapeutic response during maintenance therapy was poor in recurrent GERD, continuous therapy is recommended in order to maintain symptom-relief and mucosal healing. Hippokratia 2015, 19 (1): 53-56.
ABSTRACT
beta-Microseminoprotein was very efficiently purified from human seminal plasma with only three steps including DEAE-Sephacel and Zinc-chelate Sepharose CL-6B column chromatography. The purified protein was a non-glycoprotein with a molecular weight (M(r)) of 19,000 and 17,000 on gel filtration and reduced SDS-PAGE, respectively. The protein gave six bands from M(r) 15,600 to 25,500 on non-reduced SDS-PAGE. The characterization including the molecular weight, amino acid sequence of N-terminus and concentrations in various body fluids is discussed. Furthermore, the immunohistochemical localization of the protein among various human tissues is demonstrated.
Subject(s)
Prostatic Secretory Proteins , Proteins/chemistry , Proteins/isolation & purification , Semen/chemistry , Seminal Vesicles/metabolism , Amino Acid Sequence , Chromatography, Affinity , Chromatography, Ion Exchange , Electrophoresis, Polyacrylamide Gel , Humans , Immunohistochemistry , Male , Mercaptoethanol/chemistry , Molecular Sequence Data , Molecular Weight , Proteins/immunology , Seminal Plasma Proteins , Tissue DistributionABSTRACT
We report a 31-year-old man with facioscapulohumeral muscular dystrophy who had congenital anomalies and mental retardation. Southern blot analysis, using the probe p13E-11, displayed an abnormal EcoRI DNA fragment that reflect DNA rearrangements in facioscapulohumeral muscular dystrophy. In addition, high-resolution cytogenetic study revealed an interstitial deletion of the short arm chromosome 9: 46,XY,del(9)(p.22.1p24.1).
Subject(s)
Chromosome Deletion , Chromosomes, Human, Pair 9 , Facial Muscles , Muscular Dystrophies/genetics , Muscular Dystrophies/physiopathology , Adult , Blotting, Southern , Centromere , Chromosome Mapping , Congenital Abnormalities , Deoxyribonuclease EcoRI , Humans , Intellectual Disability , MaleABSTRACT
We found a new dysferlin gene mutation in two Japanese families, one with limb-girdle muscular dystrophy 2B and the other with Miyoshi myopathy. All patients in the limb-girdle muscular dystrophy 2B family showed apparent proximal dominant muscle atrophy and weakness, whereas a patient with Miyoshi myopathy in the second family showed distal muscle involvement at an early stage. The common clinical feature of all patients in both families was preferential involvement of calf muscles rather than the tibialis anterior muscle, which was confirmed by muscle computed tomography scan. All patients in both families shared the same homozygous alleles for chromosome 2p13 markers, and dysferlin gene analysis revealed a novel missense mutation, a G to A transition at nt 5882, which changed aspartic acid to asparagine at codon 1837. Allele-specific polymerase chain reaction analysis was used for confirmation of the mutation and for genotype analysis of the family members.
Subject(s)
Membrane Proteins , Muscle Proteins/genetics , Muscular Dystrophies/genetics , Mutation/genetics , Aged , Alleles , DNA Mutational Analysis , DNA, Complementary/analysis , DNA, Complementary/genetics , Dysferlin , Female , Haplotypes/genetics , Humans , Japan , Male , Middle Aged , Muscle Proteins/metabolism , Muscle, Skeletal/diagnostic imaging , Muscle, Skeletal/pathology , Muscle, Skeletal/physiopathology , Muscular Dystrophies/pathology , Muscular Dystrophies/physiopathology , Pedigree , Polymerase Chain Reaction , Tomography, X-Ray ComputedABSTRACT
We previously reported a variant antithrombin III (AT III Kumamoto) associated with a 31-year-old female who suffered from recurrent thrombotic episodes. To define the molecular basis for the variant AT III, we used a combination of genomic amplification followed by cloning, sequencing, and hybridization with allele-specific oligonucleotide probes. We obtained evidence for a cytosine to thymine transition in exon 2 (codon 47) of the AT III gene in the proband. This mutation converts arginine 47 to cysteine. Oligonucleotide hybridization procedures were used for confirmation of the mutation and for genotype analysis of the family members.
Subject(s)
Antithrombin III/genetics , Adult , Alleles , Base Sequence , Cloning, Molecular , DNA/genetics , Exons/physiology , Female , Gene Amplification/genetics , Genotype , Humans , Molecular Sequence Data , Mutation , Nucleic Acid Hybridization , Oligonucleotide ProbesABSTRACT
Abnormal antithrombin III (AT III) was found in the plasma of a 31-year-old female who suffered from recurrent thrombotic episodes. Heparin cofactor activity was 28% of normal and undetectable when measured by inhibition of thrombin and factor Xa (F.Xa), while both progressive antithrombin and antifactor Xa activities were normal. The concentration of plasma AT III antigen was 37 mg/dl. Analysis by crossed-immunoelectrophoresis (CIE) in the presence of heparin and affinity chromatography on heparin-Sepharose revealed that the propositus' AT III did not bind to heparin. When heparin cofactor II (HC II) was removed from propositus' plasma, heparin cofactor activity of AT III was not detected. Thus, HC II seemed to account for the plasma heparin cofactor activity found in the presence of thrombin. The patient's parents and three of her brothers demonstrated qualitative abnormality of AT III; heparin cofactor activity was 30-50% of normal levels in the presence of both thrombin and F.Xa. These findings indicate that the propositus' AT III lacks affinity for heparin and the mode of its inheritance seems to be autosomal dominant and, hence, the propositus would be a homozygote. For this variant, the name of AT III Kumamoto is proposed.
Subject(s)
Antithrombin III/genetics , Genetic Variation , Heparin/metabolism , Homozygote , Thrombosis/blood , Adult , Antithrombin III/metabolism , Blood Coagulation Tests , Chromatography, Affinity , Factor Xa , Female , Humans , Immunoelectrophoresis, Two-Dimensional , Pedigree , Serine Proteinase Inhibitors , Thrombin/antagonists & inhibitors , Thrombosis/geneticsABSTRACT
Abiesenonic acid methyl ester (AVB-I acid methyl ester), a triterpenoid compound prepared from abieslactone, suppressed tumor promoter-induced phenomena in vitro and in vivo; i.e., AVB-I acid methyl ester inhibited 12-o-tetradecanoylphorbol-13-acetate (TPA)-stimulated 32Pi-incorporation into phospholipids of cultured cells and the promoting action of TPA on skin tumor formation in mice initiated with 7,12-dimethylbenz[a]anthracene.
Subject(s)
Carcinogens/pharmacology , Lactones/pharmacology , Tetradecanoylphorbol Acetate/pharmacology , Triterpenes/pharmacology , Animals , Female , HeLa Cells/metabolism , Mice , Mice, Inbred ICR , Phospholipids/metabolism , Skin Neoplasms/chemically induced , TreesABSTRACT
Aplysiatoxin and debromoaplysiatoxin, a debrominated form of aplysiatoxin, have both been shown to be potent tumor promoters in a two-stage carcinogenesis experiment on mouse skin. However, debromoaplysiatoxin did not behave like aplysiatoxin in most of the biological assay systems using cultured cells. The discrepancy was supposed to be due to a factor in the bovine serum used for culture, a similar factor not being present in sera of eight other animal species examined. The factor was purified to homogeneity from bovine serum by ammonium sulfate fractionation and chromatographies on DEAE-cellulose, Sephadex G-150, hydroxyapatite, and a reversed-phase HPLC column. The factor was a 40-kDa protein, and partial amino-acid sequencing of its tryptic peptides indicated that the factor is alpha 1-acid glycoprotein. Both the purified factor and the commercially available bovine alpha 1-acid glycoprotein abolished in vitro the activation of protein kinase C by debromoaplysiatoxin but not that by aplysiatoxin. Debromoaplysiatoxin induced differentiation of HL-60 cells into macrophages at a comparable concentration to aplysiatoxin, when serum-free medium was used. These results suggest that alpha 1-acid glycoprotein, which interacts specifically with debromoaplysiatoxin, contained in bovine serum must have masked the in vitro properties of the tumor promoter in the biological assay systems.
Subject(s)
Anticarcinogenic Agents/isolation & purification , Carcinogens/toxicity , Lyngbya Toxins/antagonists & inhibitors , Lyngbya Toxins/toxicity , Orosomucoid/pharmacology , 3T3 Cells/drug effects , 3T3 Cells/metabolism , Amino Acid Sequence , Animals , Anticarcinogenic Agents/pharmacology , Cattle , Drug Interactions , Enzyme Activation , Epidermal Growth Factor/metabolism , Humans , Kinetics , Mice , Mice, Inbred BALB C , Molecular Sequence Data , Orosomucoid/isolation & purification , Orosomucoid/physiology , Phorbol 12,13-Dibutyrate/metabolism , Protein Kinase C/metabolism , Rabbits , Rats , Sensitivity and Specificity , Sequence Homology, Amino Acid , TritiumABSTRACT
Poly(A) polymerase [polyadenylate nucleotidyltransferase, EC 2.7.7.19] was extracted from Tetrahymena pyriformis. The enzyme was demonstrated to be present in three forms by column chromatography on DEAE-cellulose, and they were termed poly(A) polymerase Ia, Ib, and II in order of increasing affinity to the column. The properties of enzymes Ia and Ib were similar except that Ia utilizes poly(A) as a primer rather efficiently. Enzyme II differed from enzymes Ia and Ib not only in elution profile on DEAE-cellulose column chromatography but also in pH and temperature preferences, molecular weight, requirement for divalent cations, sensitivity to salts at high ionic strength, optimal primer concentration, and subcellular localization. The molecular weights of enzymes Ia and Ib measured by gel filtration were both 43,000, and that of enzyme II was 95,000. All three enzymes required Mn2+ for maximal activity; Mg2+ could replace Mn2+ in the reaction of enzyme II, but only partially. In the presence of 0.1 M ammonium sulfate the activities of enzymes Ia and Ib were both completely inhibited, whereas enzyme II still showed 42% of its original activity. These findings suggest that there are two distinct types of poly(A) polymerase in Tetrahymena pyriformis.
Subject(s)
Nucleotidyltransferases/metabolism , Polynucleotide Adenylyltransferase/metabolism , Tetrahymena pyriformis/enzymology , Animals , DNA , Isoenzymes/metabolism , Kinetics , Magnesium/pharmacology , Manganese/pharmacology , Molecular Weight , Polynucleotide Adenylyltransferase/isolation & purification , Polyribonucleotides/pharmacology , Structure-Activity RelationshipABSTRACT
Rat and mouse cDNAs for Zn-alpha 2-glycoprotein (Zn alpha 2gp) were isolated from liver libraries (lambda gt11) and compared with the human one. The lengths of cDNA inserts analyzed were 1,233 and 1,273 nucleotides for rat and mouse, respectively. The deduced amino acid sequences suggested that rat and mouse Zn alpha 2gp proteins consist of 279 and 290 amino acid residues in the mature form, respectively. They have 59.4% (rat) and 58.6% (mouse) identities in amino acid sequence with the human counterpart, and between rat and mouse the identity is 88.5%. Among the three domains, domain B is best conserved; the identities are 74.7, 73.6, and 95.6% between human and rat, human and mouse, and rat and mouse, respectively. Four cysteine and eight tryptophan residues are all conserved, and two of the three asparagine residues that carry a glycan in the human protein are conserved. Analysis of rat tissues by Northern blot suggested that its mRNA is expressed in liver, and, to a much lesser extent in submandibular gland, lung, kidney, and stomach. A more detailed study by in situ hybridization demonstrated that some epithelial cells of renal tubules and the isthmus and the neck zone cells of gastric fundic glands express Zn alpha 2gp mRNA.