ABSTRACT
STUDY QUESTION: Is there a relation between specific Na+/K+ ATPase isoform expression and localization in human blastocysts and the developmental behavior of the embryo? SUMMARY ANSWER: Na+/K+ ATPase α1, ß1 and ß3 are the main isoforms expressed in human blastocysts and no association was found between the expression level of their respective mRNAs and the rate of blastocyst expansion. WHAT IS KNOWN ALREADY: In mouse embryos, Na+/K+ ATPase α1 and ß1 are expressed in the basolateral membrane of trophectoderm (TE) cells and are believed to be involved in blastocoel formation (cavitation). STUDY DESIGN, SIZE, DURATION: A total of 20 surplus embryos from 11 patients who underwent IVF and embryo transfer at a university hospital between 2009 and 2018 were analyzed. PARTICIPANTS/MATERIALS, SETTING, METHODS: After freezing and thawing Day 5 human blastocysts, their developmental behavior was observed for 24 h using time-lapse imaging, and the expression of Na+/K+ ATPase isoforms was examined using quantitative RT-PCR (RT-qPCR). The expressed isoforms were then localized in blastocysts using fluorescent immunostaining. MAIN RESULTS AND THE ROLE OF CHANCE: RT-qPCR results demonstrated the expression of Na+/K+ ATPase α1, ß1 and ß3 isoforms in human blastocysts. Isoforms α1 and ß3 were localized to the basolateral membrane of TE cells, and ß1 was localized between TE cells. A high level of ß3 mRNA expression correlated with easier hatching (P = 0.0261). LARGE SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: The expression of mRNA and the localization of proteins of interest were verified, but we have not been able to perform functional analysis. WIDER IMPLICATIONS OF THE FINDINGS: Of the various Na+/K+ ATPase isoforms, expression levels of the α1, ß1 and ß3 mRNAs were clearly higher than other isoforms in human blastocysts. Since α1 and ß3 were localized to the basolateral membrane via fluorescent immunostaining, we believe that these subunits contribute to the dilation of the blastocoel. The ß1 isoform is localized between TE cells and may be involved in tight junction formation, as previously reported in mouse embryos. STUDY FUNDING/COMPETING INTEREST(S): This work was supported by the JSPS KAKENHI (https://www.jsps.go.jp/english/index.html), grant number 17K11215. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors have no conflicts of interest.
Subject(s)
Blastocyst , Embryo, Mammalian , Animals , Blastocyst/metabolism , Cell Membrane/metabolism , Embryo, Mammalian/metabolism , Humans , Mice , RNA, Messenger/metabolism , Sodium-Potassium-Exchanging ATPase/genetics , Sodium-Potassium-Exchanging ATPase/metabolismABSTRACT
OBJECTIVE: We examined the association between living alone and mental health and the moderating effects of face-to-face and non-face-to-face social contacts, among community-dwelling older adults. STUDY DESIGN: Cross-sectional study. METHODS: This cross-sectional study recruited Japanese adults older than 60 years, who attended health check-ups held in a suburban town hall in July and August of 2018 and 2019. As mental health outcomes, depression was assessed using the Geriatric Depression Scale 15-items, loneliness was assessed using the University of California, Los Angeles Loneliness Scale 3-items, and happiness was self-rated on a 10-point scale. Face-to-face social contacts were evaluated by participants' frequency of meetings with relatives or friends, whereas non-face-to-face contacts were measured by the frequency of interactions via letter, telephone or e-mail. Multivariable linear regression analysis was conducted to examine the association between living alone with each mental health outcome and the effect modifications of having face-to-face and non-face-to-face social contacts. RESULTS: Data from 300 older adults were analysed. The participants' mean age was 73.0 years, 51.3% were female, and 16.0% lived alone. Living alone was significantly associated with poorer mental health. Regarding loneliness and low happiness, having face-to-face and non-face-to-face contacts more than once a week alleviated the adverse association of living alone (loneliness: face-to-face contacts, P = 0.020; non-face-to-face contacts, P = 0.028; happiness: face-to-face contacts, P = 0.020; non-face-to-face contacts, P = 0.001). CONCLUSIONS: Our findings suggest that non-face-to-face, as well as face-to-face social contacts have a moderating effect on the adverse association of living alone with loneliness and happiness.
Subject(s)
Depression/epidemiology , Happiness , Independent Living/psychology , Loneliness/psychology , Social Interaction , Aged , Cross-Sectional Studies , Female , Humans , Japan/epidemiology , Male , Self ReportSubject(s)
Leukemia, Myeloid, Acute/complications , Pulmonary Alveolar Proteinosis/diagnosis , Pulmonary Alveolar Proteinosis/etiology , Aged , Bronchoalveolar Lavage Fluid , Female , Humans , Leukemia, Myeloid, Acute/physiopathology , Pulmonary Alveolar Proteinosis/diagnostic imaging , Tomography, X-Ray ComputedABSTRACT
In order to clarify the association between hyperglycemia during the early period after allogeneic stem cell transplantation (allo-SCT) and adverse outcomes, we retrospectively analyzed 563 consecutive patients who underwent allo-SCT at our institute between 2008 and 2015. Patients were categorized into three groups according to mean fasting blood glucose levels on days 0-7 (normoglycemia group<110 mg/dL, n=347; mild hyperglycemia group 110-149 mg/dL, n=192 and moderate/severe hyperglycemia group≥150 mg/dL, n=24). The median follow-up was 2.7 years. Patients in the moderate/severe hyperglycemia group had significantly worse characteristics. The cumulative incidences of 2-year non-relapse mortality (NRM) and the probabilities of 2-year overall survival (OS) in the normoglycemia, mild hyperglycemia and moderate/severe hyperglycemia groups were 7.5%, 19% and 29%, respectively (P<0.01), and 69%, 53% and 33%, respectively (P<0.01). In multivariate analyses, hyperglycemia was an independent predictor of high NRM (vs normoglycemia; mild hyperglycemia, hazard ratio (HR) 2.56, 95% confidence interval (CI) 1.56-4.18; moderate/severe hyperglycemia, HR 4.46, 95% CI 1.92-10.3) and poor OS (vs normoglycemia; mild hyperglycemia, HR 1.54, 95% CI 1.14-2.07; moderate/severe hyperglycemia, HR 1.61, 95% CI 0.89-2.91). In conclusion, hyperglycemia on days 0-7 after allo-SCT was associated with inferior outcomes.
Subject(s)
Hematopoietic Stem Cell Transplantation/adverse effects , Hyperglycemia/diagnosis , Adult , Blood Glucose/analysis , Humans , Hyperglycemia/etiology , Prognosis , Retrospective Studies , Time Factors , Transplantation, Homologous , Treatment OutcomeABSTRACT
We conducted a comprehensive analysis of 28 recurrently mutated genes in acute myeloid leukemia (AML) in 271 patients with de novo AML. Co-mutations were frequently detected in the intermediate cytogenetic risk group, at an average of 2.76 co-mutations per patient. When assessing the prognostic impact of these co-mutations in the intermediate cytogenetic risk group, overall survival (OS) was found to be significantly shorter (P=0.0006) and cumulative incidence of relapse (CIR) significantly higher (P=0.0052) in patients with complex molecular genetic abnormalities (CMGAs) involving three or more mutations. This trend was marked even among patients aged ⩽65 years who were also FLT3-ITD (FMS-like tyrosine kinase 3 internal tandem duplications)-negative (OS: P=0.0010; CIR: P=0.1800). Moreover, the multivariate analysis revealed that CMGA positivity was an independent prognostic factor associated with OS (P=0.0007). In stratification based on FLT3-ITD and CEBPA status and 'simplified analysis of co-mutations' using seven genes that featured frequently in CMGAs, CMGA positivity retained its prognostic value in transplantation-aged patients of the intermediate cytogenetic risk group (OS: P=0.0002. CIR: P<0.0001). In conclusion, CMGAs in AML were found to be strong independent adverse prognostic factors and simplified co-mutation analysis to have clinical usefulness and applicability.
Subject(s)
Chromosome Aberrations , Leukemia, Myeloid, Acute/diagnosis , Leukemia, Myeloid, Acute/genetics , Neoplasm Proteins/genetics , Adolescent , Adult , Aged , Aged, 80 and over , CCAAT-Enhancer-Binding Proteins/genetics , CCAAT-Enhancer-Binding Proteins/metabolism , Chromosomes, Human, Pair 16 , Chromosomes, Human, Pair 21 , Chromosomes, Human, Pair 8 , Cytogenetic Analysis , DNA (Cytosine-5-)-Methyltransferases/genetics , DNA (Cytosine-5-)-Methyltransferases/metabolism , DNA Methyltransferase 3A , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Dioxygenases , Female , Gene Expression , Humans , Leukemia, Myeloid, Acute/mortality , Leukemia, Myeloid, Acute/pathology , Male , Middle Aged , Neoplasm Proteins/metabolism , Nuclear Proteins/genetics , Nuclear Proteins/metabolism , Nucleophosmin , Prognosis , Proto-Oncogene Proteins/genetics , Proto-Oncogene Proteins/metabolism , Recurrence , Retrospective Studies , Survival Analysis , fms-Like Tyrosine Kinase 3/genetics , fms-Like Tyrosine Kinase 3/metabolismABSTRACT
Gene mutations were found in acute myeloid leukemia (AML) and their importance has been noted. To clarify the importance and stability of mutations, we examined gene mutations in paired samples at diagnosis and relapse of 34 adult AML patients. Five acquired gene mutations were detected at relapse. Of the 45 gene mutations at diagnosis, 11 of them were lost at relapse. The acquired mutations at relapse were all class I mutations as Fms-like tyrosine kinase 3 (FLT3) and rat sarcoma viral oncogene homolog (RAS) mutations. The disappeared mutations at relapse were 3 of 11 internal tandem duplications of FLT3 (FLT3-ITD) (27.3%), 3 of 3 FLT3 tyrosine kinase domain (FLT3-TKD) (100%), 3 of 13 Nucleophosmin 1 (23.1%) and 2 of 5 CCAAT/enhancer-binding protein-α (40%) mutations. However, epigenetics-modifying gene (DNMT3a, TET2 and IDH1/2) mutations had no change between diagnosis and relapse samples, and may become minimal residual disease marker. The frequency of FLT3-ITD at relapse in patients with DNMT3a mutation at diagnosis is significantly higher than those in patients without them (P=0.001). Moreover, the high frequency of FLT3-ITD at relapse is also seen in AML cases that initially present with any epigenetics-modifying gene mutations (P<0.001). Our results indicate that epigenetics-modifying gene mutations may cause genetic instability and induce FLT3-ITD, leading to resistance to therapy and relapse.
Subject(s)
DNA (Cytosine-5-)-Methyltransferases/genetics , DNA-Binding Proteins/genetics , Isocitrate Dehydrogenase/genetics , Leukemia, Myeloid, Acute/genetics , Mutation , Proto-Oncogene Proteins/genetics , fms-Like Tyrosine Kinase 3/genetics , DNA Methyltransferase 3A , Dioxygenases , Epigenomics , Humans , Nucleophosmin , RecurrenceABSTRACT
AIMS: Squamous cell carcinoma (SCC) is the most common form of malignant transformation in mature cystic teratoma (MCT) of the ovary. Some investigators have suggested the possibility of origin from columnar epithelium. The aim of this study was to analyse such tumours immunohistochemically to elucidate their histogenesis. METHODS AND RESULTS: The expression of cytokeratin (CK) 10 and CK18 was examined in 21 samples of SCC arising in MCT. The expression of CK10 and CK18 was also assessed in SCCs arising in different organs (skin, vulva, lung and uterine cervix) for the purpose of comparison. SCC in MCT expressed CK10 in 7/21 cases [33.3%, 95% confidence interval (CI) 0.12-0.53] and CK18 in 14/21 cases (66.7%, 95% CI 0.46-0.87). SCC in MCT expressed CK10 less frequently, but CK18 more frequently, as is the case in SCCs of the uterine cervix (CK10, 20%; CK18, 70%) and the lung (CK10, 5%; CK18, 90%), both of which are derived from columnar epithelium by squamous metaplasia. CONCLUSIONS: SCC in MCT may be derived from metaplastic squamous epithelium.
Subject(s)
Carcinoma, Squamous Cell/pathology , Keratin-18/metabolism , Ovarian Neoplasms/pathology , Teratoma/pathology , Adult , Aged , Aged, 80 and over , Carcinoma, Squamous Cell/metabolism , Cell Transformation, Neoplastic/metabolism , Epithelium/metabolism , Epithelium/pathology , Female , Gene Expression Regulation, Neoplastic , Humans , Keratin-10/genetics , Keratin-10/metabolism , Keratin-18/genetics , Middle Aged , Ovarian Neoplasms/metabolism , Ovary/metabolism , Ovary/pathology , Teratoma/metabolismABSTRACT
OBJECTIVE: Early cervical adenocarcinoma (ECA) with a tumour depth of <3 mm has a good prognosis. To clarify the cytological features of ECAs with depth <3 mm, these were compared with those of ECA with 3-5 mm and invasive adenocarcinoma (IA) invading the cervical wall with more than 5 mm in depth. METHODS: The cervical cytological features of ECAs with depth <3 mm (14 cases) were compared with those of ECA with 3-5 mm (four cases) and IA (13 cases). Cytologically, the presence or absence of tumour diathesis, number of atypical cells, crowded cell groups, groups with glandular structures, feathering, groups with palisading borders, rosettes, clusters, cell shape and size, nuclear shape and size, nucleolar shape and size, chromatin distribution, border and character of cytoplasm, and single cell pattern were evaluated. RESULTS: A tumour diathesis was seen in five of 14 ECA <3 mm in depth (36%), all four ECA with 3-5 mm (100%) and 11 of 13 IA with more than 5 mm (85%). Single cells, macronucleoli and coarsely granular chromatin pattern were less frequent in ECA of <3 mm than that in ECA with 3-5 mm and IA. The number of atypical cells and glandular structures in ECA was significantly less than that in IA. Cell crowding, feathering, palisading and rosettes were common in both ECA and IA. CONCLUSION: The characteristic cytological features of ECA with depth <3 mm, having a good prognosis, were clean background, fewer single cells and macronucleoli, and less frequent coarsely granular chromatin pattern compared with those in ECA with 3-5 mm and IA. The number of atypical cells and glandular structures in ECA was significantly less than that in IA. Familiarity with the cytological features of ECA and its mimics is essential.
Subject(s)
Adenocarcinoma/diagnosis , Uterine Cervical Neoplasms/diagnosis , Cervix Uteri/pathology , Early Diagnosis , Female , Humans , Lymphatic Metastasis , Neoplasm Invasiveness , Neoplasm Recurrence, Local , PrognosisABSTRACT
OBJECTIVE: The aim of this study was to determine whether or not we could distinguish uterine papillary serous carcinoma (UPSC) from other types of endometrial cancer by cytology. METHODS: We examined the cytological findings of the endometrium from five cases with UPSC and compared them with 10 cases with endometrioid adenocarcinoma, grade 1 (G1). A morphometric analysis was performed. Cytological samples from the cervix and ascites of the patients with UPSC were also reviewed. RESULTS: All five patients had FIGO stage III and IV tumours. Three patients died of the disease and two are still alive with disease. The tumour cells of UPSC tended to be shed in papillary clusters with a tumour diathesis. Psammoma bodies were seen only in UPSC. The frequency of irregular-shaped nuclei, membrane thickness and eccentric nuclei in UPSC was higher than in G1. The chromatin pattern was coarsely granular, and both anisonucleosis and bare nuclei were prominent in UPSC. Cytomorphometrically, the maximum diameter of the nuclei in UPSC was significantly greater than that in G1. The nucleoli were also more often seen in UPSC than in G1. The findings of the nuclei and nucleoli in the cervical and peritoneal fluid cytology closely resembled those in endometrial smears. The features of the cervical smears and peritoneal fluid cytology were different from those of endometrial cytology regarding clear background and small clusters of cells. CONCLUSION: As the endometrial cytology findings accurately suggested the histological diagnosis of UPSC, the diagnosis of UPSC was confirmed in this study by endometrial cytology. The cytological diagnosis of UPSC should be based on the findings of tumour diathesis, psammoma bodies and papillary clusters composed of tumour cells with enlarged nuclei and numerous nucleoli.
Subject(s)
Cystadenocarcinoma, Papillary/pathology , Endometrial Neoplasms/pathology , Aged , Cystadenocarcinoma, Papillary/diagnosis , Endometrial Neoplasms/diagnosis , Female , Humans , Middle Aged , Neoplasm StagingABSTRACT
We investigated the energetic costs of quadrupedal and bipedal walking in two Japanese macaques. The subjects were engaged in traditional bipedal performance for years, and are extremely adept bipeds. The experiment was conducted in an airtight chamber with a gas analyzer. The subjects walked quadrupedally and bipedally at fixed velocities (<5 km/hr) on a treadmill in the chamber for 2.5-6 min. We estimated energy consumption from carbon dioxide (CO2) production. While walking bipedally, energetic expenditure increased by 30% relative to quadrupedalism in one subject, and by 20% in another younger subject. Energetic costs increased linearly with velocity in quadrupedalism and bipedalism, with bipedal/quadrupedal ratios remaining almost constant. Our experiments were relatively short in duration, and thus the observed locomotor costs may include presteady-state high values. However, there was no difference in experimental duration between bipedal and quadrupedal trials. Thus, the issue of steady state cannot cancel the difference in energetic costs. Furthermore, we observed that switching of locomotor mode (quadrupedalism to bipedalism) during a session resulted in a significant increase of CO2 production. Taylor and Rowntree ([1973] Science 179:186-187) noted that the energetic costs for bipedal and quadrupedal walking were the same in chimpanzees and capuchin monkeys. Although the reason for this inconsistency is not clear, species-specific differences should be considered regarding bipedal locomotor energetics among nonhuman primates. Extra costs for bipedalism may not be great in these macaques. Indeed, it is known that suspensory locomotion in Ateles consumes 1.3-1.4 times as much energy relative to quadrupedal progression. This excess ratio surpasses the bipedal/quadrupedal energetic ratios in these macaques.
Subject(s)
Energy Metabolism/physiology , Gait/physiology , Macaca/physiology , Walking/physiology , Animals , Carbon Dioxide/metabolism , Male , RespirationABSTRACT
The placental trophoblast is considered to act as a barrier between mother and fetus, mediating the exchange of various materials across the placenta. ATP-binding cassette (ABC) transporters such as P-glycoprotein (P-gp) and multidrug-resistance protein (MRP) are expressed in the placenta and function as efflux transport systems for xenobiotics. In the present study, we aimed to determine the localization of MRP1 in the human placenta in comparison with that of P-gp. Western blotting analysis with human placental membrane vesicles indicated that P-gp and MRP1 are localized on the brush-border membranes and basal membranes, respectively. Immunohistochemical analysis with human normal full-term placenta showed that anti-P-gp monoclonal antibody F4 stained the brush-border side of the trophoblast cells, whereas anti-MRP1 monoclonal antibody MRPr1 stained the basal side. These results confirm that P-gp and MRP1 are located on the brush-border membranes and basal membranes, respectively, of human full-term placental trophoblast. MRP1 was also detected on the abluminal side of blood vessels in the villi. Accordingly, MRP1 may play a role distinct from that of P-gp, which is considered to restrict the influx of xenobiotics into the fetus.
Subject(s)
Multidrug Resistance-Associated Proteins/analysis , Trophoblasts/chemistry , ATP Binding Cassette Transporter, Subfamily B/analysis , ATP Binding Cassette Transporter, Subfamily B/metabolism , Basement Membrane/chemistry , Basement Membrane/cytology , Blotting, Western , Female , Humans , Immunohistochemistry , Multidrug Resistance-Associated Proteins/immunology , Placenta/cytology , Placenta/metabolism , Pregnancy , Transport Vesicles/chemistry , Trophoblasts/cytology , Trophoblasts/metabolismABSTRACT
RCAS1, which acts as a ligand for a putative receptor on immune cells such as peripheral lymphocytes and natural killer cells, is strongly expressed in human cancers. RCAS1 can induce these cells to undergo apoptotic cell death, which suggests that RCAS1 expression may prohibit the stromal reaction occurring in a tumour. To clarify the clinical significance of RCAS1 expression in uterine endometrial cancer, we analysed the association between RCAS1 expression and clinicopathologic variables by statistical methods. With the use of immunohistochemical techniques, we performed a retrospective study of RCAS1 expression in resected tumour tissue from 147 patients with uterine endometrial cancer. We evaluated the statistical correlation between RCAS1 expression and clinicopathologic variables. RCAS1 was expressed in 106 of 147 patients with uterine endometrial cancer; 30 of these 147 patients showed RCAS1 overexpression. Overexpression of RCAS1 was significantly correlated with age at surgery, stage, extent of myometrial invasion, and positive peritoneal cytologic results. Multivariate analysis revealed that RCAS1 expression and metastasis were clinically significant prognostic factors for the overall survival. These findings indicated that analysis for RCAS1 expression can provide crucial information about the clinical behaviour of uterine endometrial cancer, which may be valuable for the management of patients with this disease.
Subject(s)
Antigens, Neoplasm/biosynthesis , Endometrial Neoplasms/genetics , Endometrial Neoplasms/pathology , Gene Expression Regulation, Neoplastic , Neoplasm Invasiveness , Neoplasm Metastasis , Adult , Aged , Aged, 80 and over , Antigens, Neoplasm/analysis , Female , Humans , Middle Aged , Multivariate Analysis , Prognosis , Survival Analysis , Treatment OutcomeABSTRACT
PURPOSE: The aim of the present study was to establish the accurate cutoff points of post-treatment serum beta-hCG values in identifying chemotherapeutic refractory cases among patients with low-risk persistent trophoblastic disease (PTD) treated with 8-day methotrexate-folinic acid as the primary therapy. MATERIALS AND METHODS: The values of serum beta-hCG measured before initiating treatment and weekly thereafter in 26 patients with low-risk PTD undergoing 8-day methotrexate-folinic acid treatment were analyzed. Thereafter, we determined the weekly cutoff points to identify the patient refractory for treatment by means of receiver-operating characteristic (ROC) plots analysis. RESULTS: The values of cutoff points in the pretreatment, the post-treatment 1st, 2nd, 3rd, and 4th week were 18.6, 15.0, 5.4, 3.4, and 2.0 ng/ml, respectively, and the value of accuracy during these weeks was appropriate (> 80%). When using the cutoff points of one and two weeks after initiating treatment, the accuracy in identifying chemotherapeutic refractory patients was 87.5% and 88.0%, respectively, with the highest values exceeding 85%. The sensitivity and specificity at one week were 92.9 and 80.0%, respectively. Similarly, the sensitivity and specificity at two weeks were 93.3 and 80.0%, respectively. CONCLUSION: These results suggest that the cutoff points of one and two weeks after initiating treatment are useful in identifying chemotherapeutic refractory patients among low-risk PTD patients, receiving 8-day methotrexate-folinic acid treatment.
Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Biomarkers, Tumor/blood , Chorionic Gonadotropin/blood , Leucovorin/administration & dosage , Lung Neoplasms/drug therapy , Methotrexate/administration & dosage , Trophoblastic Neoplasms/drug therapy , Uterine Neoplasms/drug therapy , Antimetabolites, Antineoplastic/administration & dosage , Female , Humans , Lung Neoplasms/blood , Pregnancy , Reference Values , Sensitivity and Specificity , Treatment Outcome , Trophoblastic Neoplasms/blood , Uterine Neoplasms/bloodABSTRACT
The hydroxyl radical (*OH) is generated in polluted dew on the needle surfaces of Japanese red pine (Pinus densiflora Sieb. et Zucc.). This free radical, which is a potent oxidant, is assumed to be a cause of ecophysiological disorders of declining trees on the urban-facing side of Mt. Gokurakuji, western Japan. Mists of *OH-generating N(III) (HNO2 and NO2-) and HOOH + Fe + oxalate solutions (50 and 100 microM, pH 5.1-5.4) simulating the dew water were applied to the foliage of pine seedlings grown in open-top chambers in the early morning. Needles treated with 100 microM N(III) tended to have a greater maximum CO2 assimilation rate (Amax), a greater stomatal conductance (g(s)) and a greater needle nitrogen content (Nneedle), suggesting that N(III) mist acts as a fertilizer rather than as a phytotoxin. On the other hand, needles treated with 100 microM HOOH + Fe + oxalate solution showed the smallest Amax, g(s), and Nneedle, suggesting that the combination of HOOH + Fe + oxalate caused a decrease in needle productivity. The effects of HOOH + Fe + oxalate mist on pine needles were very similar to the symptoms of declining trees at Mt. Gokurakuji.
Subject(s)
Carbon Dioxide/metabolism , Photosynthesis/physiology , Pinus/physiology , Carbon Dioxide/analysis , Cities , Hydroxyl Radical/chemistry , Iron/chemistry , Nitrogen/metabolism , Oxalates/chemistry , Plant Leaves/chemistry , Population Dynamics , Water/chemistryABSTRACT
Ubiquitin-conjugated proteins in human colorectal cancer tissues were analyzed by the immunoprecipitation with the antibody FK2 against conjugated ubiquitin followed with SDS-PAGE. In these immunoprecipitable proteins, a 38-kDa protein was abundant in the tumor regions but almost absent in the adjacent normal regions in 17/26 patients, thus we attempted to purify it. Using immunoaffinity chromatography with the antibody FK2 followed by gel filtration and SDS-PAGE, approximately 10 pmol of this protein was separated from 34 g of the pooled cancerous tissue and transferred onto a PVDF membrane. The 38-kDa protein was further digested with Achromobacter protease I, resulting in several peptide fragments. Amino acid sequences of these peptides showed complete sequence identity to those derived from either ubiquitin or phosphoglycerate mutase-B, suggesting that the 38-kDa protein is monoubiquitinated phosphoglycerate mutase-B, whose calculated mass is 37,369 Da. Western blot using an antibody against phosphoglycerate mutase-B revealed the presence of the 38-kDa protein in the anti-ubiquitin immunoprecipitates derived from the tumor regions, but not from normal counterparts. In addition, part of non-ubiquitinated phosphoglycerate mutase-B (29 kDa) was also found in the anti-ubiquitin immunoprecipitates, whose levels were higher in the tumor regions than in the adjacent normal regions. These results suggest that monoubiquitination of phosphoglycerate mutase-B as well as formation of a noncovalent complex containing ubiquitin and phosphoglycerate mutase-B increases in colorectal cancer and novel modification of phosphoglycerate mutase-B might have a pathophysiological role.
Subject(s)
Colorectal Neoplasms/enzymology , Phosphoglycerate Mutase/isolation & purification , Ubiquitin/metabolism , Aged , Aged, 80 and over , Amino Acid Sequence , Chromatography, Affinity , Female , Humans , Male , Middle Aged , Molecular Sequence Data , Molecular Weight , Phosphoglycerate Mutase/chemistryABSTRACT
BACKGROUND: This study used the clinicopathologic profiles of Japanese women younger than 50 years of age with endometrial carcinoma to distinguish the clinicopathologic features of carcinomas of the lower uterine segment (LUS) from those of carcinomas of the corpus mucosa proper (CMP). METHODS: Eighty-eight endometrial carcinomas in women younger than 50 years old (25.3%) were selected from our file of 348 Japanese women with endometrial carcinoma. Seventy-two were classified as carcinomas of the CMP and 16 carcinomas of the LUS. A tumor was judged to be a carcinoma of the LUS when it involved a continuous area ranging from the lower corpus to the upper cervix with or without intervention of a portion of uninvolved LUS. RESULTS: The mean ages of women with carcinomas of the CMP and LUS were 41.2 and 39.0 years, respectively. In comparison to carcinomas of the LUS, carcinomas of the CMP were more strongly associated with reproductive risk factors including parity (P = 0.01) and polycystic ovary syndrome (P = 0.02). There was no significant difference in body mass index or the incidence of diabetes mellitus and hypertension between women presenting with carcinomas of the CMP and LUS. Histologically, carcinomas of the LUS more frequently showed a high-grade endometrioid tumor (P = 0.02) with deep myometrial invasion (P < 0.01) and were less associated with endometrial hyperplasia (P < 0.01) than those of the CMP. CONCLUSIONS: Carcinomas of the LUS occurred predominantly in women younger than 50 years of age and had clinicopathologic features distinct from carcinomas of the CMP in women younger than 50 years of age.
Subject(s)
Adenocarcinoma/pathology , Endometrial Neoplasms/pathology , Neoplasm Invasiveness , Adolescent , Adult , Age Factors , Body Mass Index , Diabetes Complications , Female , Humans , Hypertension/complications , Middle Aged , Parity , Polycystic Ovary Syndrome/complications , Retrospective Studies , Risk FactorsABSTRACT
We show that most of the internalized rat LH receptor is routed to a lysosomal degradation pathway whereas a substantial portion of the human LH receptor is routed to a recycling pathway. Chimeras of these two receptors identified a linear amino acid sequence (GTALL) present near the C terminus of the human LH receptor that, when grafted onto the rat LH receptor, redirects most of the rat LH receptor to a recycling pathway. Removal of the GTALL sequence from the human LH receptor failed to affect its routing, however. The GTALL sequence shows homology with the C-terminal tetrapeptide (DSLL) of the beta2-adrenergic receptor, a motif that has been reported to mediate the recycling of the internalized beta2-adrenergic receptor by binding to ezrin-radixin-moesin-binding phosphoprotein-50. Addition of the DSLL tetrapeptide to the C terminus of the rat LH receptor also redirects most of the internalized rat LH receptor to a recycling pathway but, like the recycling of the human LH receptor, this rerouting is not mediated by ezrin-radixin-moesin-binding phosphoprotein-50. We conclude that most of the internalized rat LH receptor is degraded because its C-terminal tail lacks motifs that promote recycling and that two distinct, but homologous, motifs (DSLL at the C terminus or GTALL near the C terminus) can reroute the internalized rat LH receptor to a recycling pathway that is independent of ezrin-radixin-moesin-binding phosphoprotein-50.
Subject(s)
Endocytosis/physiology , Receptors, LH/chemistry , Receptors, LH/metabolism , Amino Acid Motifs , Amino Acid Sequence , Animals , Chorionic Gonadotropin/metabolism , Humans , Molecular Sequence Data , Rats , Receptors, LH/genetics , Recombinant Fusion Proteins/chemistry , Recombinant Fusion Proteins/metabolism , TransfectionABSTRACT
OBJECTIVE: To clarify the relationship between age, histological type, and size of ovarian tumors. METHOD: A review was made of 1648 cases of histopathologically diagnosed ovarian tumors and tumor-like lesions, and information on the age of the patients and size of the tumor was obtained. Statistical analysis was performed using Kruskal-Wallis tests or Mann-Whitney U-tests. RESULTS: There were 840 (51%) cases of benign tumors, 73 (4%) cases of tumors of low malignant potential (LMP), 268 (16%) cases of malignant tumors and 467 (28%) cases of tumor-like lesions. The age of the patients was significantly different among tumor-like lesions (34.6+/-8.1 years), benign tumors (39.8+/-16.4 years), LMP tumors (45.2+/-18.3 years) and malignant tumors (51.9+/-13.0 years) (P<0.0001). The maximum diameter of the tumors was significantly different among tumor-like lesions (7.1+/-3.3 cm), benign tumors (10.9+/-5.6 cm), malignant tumors (13.6+/-6.5 cm) and LMP tumors (18.5+/-6.8 cm) (P<0.0001). CONCLUSION: The distribution of tumor histological type (tumor-like lesions, benign, LMP and malignant) was correlated against patient age and lesion diameter, with tumors in older patients or larger tumors more likely to be malignant.