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1.
Gan To Kagaku Ryoho ; 51(2): 205-207, 2024 Feb.
Artigo em Japonês | MEDLINE | ID: mdl-38449414

RESUMO

Advanced breast cancer with skin invasion are relatively often accompanied by bleeding, effusion, malodor, and pain, which reduce the quality of life(QOL)of the patients and their families. Therefore, local symptom control is as important as surgical treatment and chemotherapy. We control these symptoms by using cadexomer iodine, cadexomer iondine is expensive, but relatively easy for family to use and keep patients' QOL.


Assuntos
Neoplasias da Mama , Humanos , Feminino , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/cirurgia , Qualidade de Vida , Iodóforos , Pele
2.
BMC Womens Health ; 23(1): 620, 2023 11 21.
Artigo em Inglês | MEDLINE | ID: mdl-37990218

RESUMO

BACKGROUND: Recently, there has been a growing global movement concerning menstruation, a healthy and natural physiological phenomenon in women. The disadvantages caused by menstruation are "gender-based obstacles." Adolescent girls are also under its influence and perhaps in a more vulnerable situation than adult women. This study investigated the experiences related to menstruation that affect health-related quality of life (HRQOL) of high school students in Japan. METHODS: This cross-sectional study was conducted at a municipal high school in Fukuoka Prefecture, Japan. The study population comprised 233 female students among which 198 completed the questionnaire. The questionnaire contained items about menstruation and HRQOL measured by the 36-Item Short Form Health Survey (SF-36). RESULTS: Approximately a quarter had experienced difficulties in obtaining sanitary products in the past year, whether for economic or non-economic reasons. Menstruation-associated symptoms, impact on daily life, trouble with menstruation at an unexpected time, usage of painkillers, unhealthy lifestyle, and negative perception of menstruation were significantly associated with lower HRQOL scores, particularly in the mental component summary scores of the SF-36. CONCLUSIONS: For the high school students with severe menstruation-associated symptoms that interfere with their daily lives, the results of this study suggest that improving access to medical care, information, and education can contribute to a better HRQOL.


Assuntos
Menstruação , Qualidade de Vida , Adolescente , Feminino , Humanos , Estudos Transversais , População do Leste Asiático , Conhecimentos, Atitudes e Prática em Saúde , Estudantes , Inquéritos e Questionários
3.
Oncol Lett ; 25(4): 139, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36909373

RESUMO

Trefoil factors (TFFs) are upregulated in numerous types of cancer, including those of the breast, the colon, the lung and the pancreas, suggesting their potential utility as biomarkers for screening. In the present study, the clinical relevance of serum or urinary TFFs as biomarkers were comprehensively evaluated and the correlation with TFF expression levels in lung cancer tissue was examined. Serum and urine were collected from 199 patients with lung cancer and 198 healthy individuals. Concentrations of serum and urinary TFF1, TFF2 and TFF3 were measured using ELISA and the potential of TFF levels to discriminate between cancer and non-cancer samples was evaluated. In 100 of the cancer cases, expression of TFF1-3 was analyzed using immunohistochemical staining of paraffin sections. Furthermore, the relationship between TFF levels and clinicopathological factors among these cancer cases was analyzed using immunohistochemistry of tissue specimens, quantified and statistically analyzed. While serum levels of all TFFs measured using ELISA were significantly higher in patients with lung cancer compared with those in healthy individuals, urinary TFFs were lower. Areas under the curve (AUC) of the receiver operating characteristic curves for serum/urinary TFF1, TFF2 and TFF3 were 0.709/0.594, 0.722/0.501 and 0.663/0.665, respectively. Furthermore, the combination of serum TFF1, TFF2, TFF3 and urinary TFF1 and TFF3 demonstrated the highest AUC (0.826). In the clinicopathological analysis, serum TFF1 was higher in the early pathological T-stage (pTis/1/2) compared with the later stage (pT3/4) and TFF2 was higher in the pN0/1 than the pN2 group. With regards to the histological types, urinary TFF1 was higher in squamous cell carcinoma than adenocarcinoma (AC), but TFF2 tended to be higher in AC. Using immunohistochemical analysis, although TFF1 and TFF3 expression showed positive correlation with serum concentrations, TFF2 was inversely correlated. In conclusion, serum and urinary TFF levels are promising predictive biomarkers, and their measurements provide a useful in vivo and non-invasive diagnostic screening tool. In particular, TFF1 and TFF3 could be surrogate markers of clinicopathological profiles of human lung cancer.

4.
Micromachines (Basel) ; 13(6)2022 May 27.
Artigo em Inglês | MEDLINE | ID: mdl-35744451

RESUMO

The quantitative and functional analyses of cells are important for cell-based therapies. In this study, to establish the quantitative cell analysis method, we propose an impedance measurement method supported by dielectrophoretic cell accumulation. An impedance measurement and dielectrophoresis device was constructed using opposing comb-shaped electrodes. Using dielectrophoresis, cells were accumulated to form chain-like aggregates on the electrodes to improve the measurement sensitivity of the electrical impedance device. To validate the proposed method, the electrical impedance and capacitance of primary and de-differentiated chondrocytes were measured. As a result, the impedance of the chondrocytes decreased with an increase in the passage number, whereas the capacitance increased. Therefore, the impedance measurement method proposed in this study has the potential to identify chondrocyte phenotypes.

5.
Sci Rep ; 12(1): 4811, 2022 03 21.
Artigo em Inglês | MEDLINE | ID: mdl-35314746

RESUMO

Genetic analysis and culturing techniques for gastric non-Helicobacter pylori Helicobacter (NHPH) are progressing. NHPH is reported to accompany nodular gastritis, gastric MALT lymphoma, and mild gastritis. However, only a few gastric cancer cases infected by NHPH have been reported. PCR analysis specific for NHPH and H. pylori was performed for DNA from gastric mucosa of 282 Korean gastric cancer patients, who were treated with endoscopic submucosal dissection. For more precise strain detection of NHPH, NHPH-positive mucosa was stained by immunohistochemistry specific for Helicobacter suis. The Cancer Genome Atlas (TCGA) classification was analyzed for these 3 gastric cancer sub-groups by in situ hybridization and immunohistochemistry. Among 281 patients, 3 patients (1.1%) were positive for NHPH. One patient (Patient 1) was also positive for H. pylori by PCR, another patient (Patient 3) was positive for serum IgG for H. pylori, and the other patient (Patient 2) had no evidence for H. pylori infection. Gastric mucosa of Patients 2 and 3 were positive for H. suis staining. All three NHPH-positive gastric cancers were located in the antrum, and belonged to the Chromosomal Instability Type of TCGA classification. Gastric NHPH can be a cause of gastric cancer, although likely with lower pathogenesis than H. pylori.


Assuntos
Gastrite , Infecções por Helicobacter , Helicobacter pylori , Helicobacter , Neoplasias Gástricas , Mucosa Gástrica/patologia , Gastrite/patologia , Infecções por Helicobacter/patologia , Helicobacter pylori/genética , Humanos , Neoplasias Gástricas/patologia
6.
Gan To Kagaku Ryoho ; 49(13): 1461-1463, 2022 Dec.
Artigo em Japonês | MEDLINE | ID: mdl-36733102

RESUMO

CREATE-X trial demonstrated the effectiveness of additional capecitabine therapy in prolonging disease-free survival among patients who are HER2 negative, especially those with triple-negative breast cancer who had residual invasive disease after standard neoadjuvant chemotherapy. We investigated our data regarding adjuvant capecitabine for residual disease. Ten patients were enrolled, and the average age of the patients was 54.2 years. All patients completed 8 courses of treatment; all adverse events were Grade 2 or lower. Five-year disease-free survival rate was 70.0% in an average observation period of 40.9 months. Three patients recurred within 2 years, and all patients had brain metastasis. In the CREATE-X trial, the rate of disease-free survival was 69.8% in the capecitabine group versus 56.1% in the control group; our results were same as those of CREATE-X. Brain metastasis may be detected by the early phase of enhanced brain MRI.


Assuntos
Neoplasias da Mama , Neoplasias de Mama Triplo Negativas , Humanos , Pessoa de Meia-Idade , Feminino , Capecitabina/efeitos adversos , Neoplasias de Mama Triplo Negativas/tratamento farmacológico , Neoplasias de Mama Triplo Negativas/cirurgia , Terapia Neoadjuvante , Neoplasias da Mama/tratamento farmacológico , Quimioterapia Adjuvante/métodos , Recidiva Local de Neoplasia/tratamento farmacológico , Intervalo Livre de Doença , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico
7.
Onco Targets Ther ; 14: 4761-4777, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34531663

RESUMO

INTRODUCTION: Trefoil Factor (TFF) is a member of a protein family comprised of three isoforms, of which TFF-1 exhibits antithetical functions; promotion or suppression of cell proliferation, survival and invasion, depending on the cancer type. However, the pathobiological function of TFF-1 in lung carcinoma has been still unclear. METHODS: We examined the expression and secretion of TFF-1 using cultured human lung carcinoma cells by immunoblotting, immunofluorescence, enzyme-linked immunosorbent assay and quantitative real-time PCR analyses. The effects of TFF-1 on various phenotypes were analyzed in two cell lines, including those transfected with cDNA encoding TFF-1. Cell proliferation and death were examined by hemocytometer cell counting and by colorimetric viability/cytotoxicity assay. Cell cycle profile, migration and invasion were also examined by flow cytometry, wound healing assay and Matrigel Transwell assay, respectively. The effect of TFF-1 overexpression was confirmed by additional transfection of TFF-1-specific siRNA. RESULTS: Endogenous TFF-1 protein expression and secretion into the media were observed exclusively in adenocarcinoma-derived cell lines. Forced overexpression of TFF-1 drove cell cycle transition, while the proliferation decreased by 19% to 25% due to increased cell death. This cell death was predominantly caused by apoptosis, as assessed by the activation of caspase 3/7. Cell migration was also suppressed by 71% to 82% in TFF-1-transfected cells. The suppressive effect of TFF-1 on proliferation and migration was restored by transfection of TFF-1 siRNA. Moreover, invasion was also suppressed to 77% to 83% in TFF-1-transfected cells. CONCLUSION: These findings reveal that TFF-1 functions as a suppressor of cancer proliferation by induction of apoptosis, cell migration and invasion and thus may provide a synergistic target for potential treatment strategies for human lung carcinoma.

8.
Case Rep Oncol ; 14(2): 944-949, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34248562

RESUMO

Spina bifida (SB) is a congenital neural tube defect that often presents with neurological disability and decubitus ulcers. A 66-year-old woman with SB presented to our hospital with decubitus ulcers and was treated by a plastic surgeon. She was referred to our department because of a mass measuring 5 × 4 cm in the superolateral quadrant of the right breast. The size of the right axillary lymph node (LN) was 2 × 1 cm. A core-needle biopsy revealed an invasive ductal carcinoma. Total mastectomy and axillary LN dissection were planned. However, 2 days prior to surgery, the size of the mass and the LN rapidly increased to 7 × 4 cm and 3 × 2 cm, respectively. Furthermore, the enlarged LN was close to the thoracodorsal artery. Since complete resection was difficult, neoadjuvant chemotherapy was also administered. On day 11 of neoadjuvant chemotherapy, the patient was febrile and developed a decubitus ulcer infection at the buttock. The neutrophil count was within normal limits; thus, she was not diagnosed with febrile neutropenia. Follow-up computed tomography revealed a shrinking of the mass to 5 × 4 cm after the first cycle of neoadjuvant chemotherapy. After 17 days of antibiotic therapy and drainage, total mastectomy and axillary LN dissection were performed. Due to the risk of recurrence of infection, adjuvant chemotherapy was discontinued and hormone therapy was initiated. In conclusion, indications for chemotherapy should be carefully evaluated in SB patients with lower limb paralysis and decubitus ulcers.

9.
Shokuhin Eiseigaku Zasshi ; 62(3): 100-104, 2021.
Artigo em Japonês | MEDLINE | ID: mdl-34219097

RESUMO

A microwave digestion method for the analysis of tin, arsenic and lead in soft drinks was studied and its performance was evaluated. The trueness, repeatability (RSD) and Intralaboratory reproducibility (RSD) of the method were estimated to be in the ranges of 93-100, 0.7-6.1 and 0.9-8.6%, respectively. In the preparation of the samples for tin analysis, the addition of sulfuric acid after microwave digestion improved the recovery rate and reproducibility. To verify the applicability of this method, recovery tests were conducted on five types of soft drinks, and the results were satisfactory. It was confirmed that this method is applicable to the digestion of a wide range of soft drinks and to the analysis of tin, arsenic, and lead, and that it reduces working time and makes the work easier compared to the conventional method.


Assuntos
Arsênio , Arsênio/análise , Bebidas Gaseificadas , Digestão , Micro-Ondas , Reprodutibilidade dos Testes , Estanho
10.
Pharmaceutics ; 13(2)2021 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-33562278

RESUMO

Ghrelin is the peptide that increases the hunger sensation and food intake and is expected to be clinically applied for treatment of diseases such as cachexia and anorexia nervosa. In the clinical application of ghrelin, injections are problematic in that they are invasive and inconvenient. Thus, we aimed to develop a formulation that can eliminate the need for injections and can be applied clinically. We prepared formulations of an hGhrelin derivative, in which the octanoyl group essential for expression of activity is modified to avoid rapid des-acylation, using lyophilizate for a dry powder inhalation (LDPI) system. The formulation of hGhrelin derivative was optimized by the addition of phenylalanine, of which the fine particle fraction of 5 µm or less was 41.7 ± 3.8%. We also performed pharmacokinetic/pharmacodynamic tests in monkeys using the optimum formulation that can be applied clinically. The absolute bioavailability of inhaled hGhrelin derivative with respect to that intravenously injected was 16.9 ± 2.6%. An increase in growth hormone was shown as an effect of the inhaled hGhrelin derivative similar to intravenous injection. The LDPI formulation can deliver the hGhrelin derivative systemically, and it is expected to be applied clinically as a substitute for injections.

11.
Surg Case Rep ; 5(1): 157, 2019 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-31654286

RESUMO

BACKGROUND: Angiosarcoma of the breast is rare. It carries a poor prognosis because of its high risk of local recurrence and distant metastases. Presently, there are still no established systemic therapies. Thus, the main treatment strategy for breast angiosarcoma is complete resection. This underscores the importance of closely monitoring the spread of the tumor lesion, particularly for multifocal angiosarcoma, and to plan an optimal operative procedure. We herein present the successful surgical treatment of a rare case of multifocal primary breast angiosarcoma. CASE PRESENTATION: A 43-year-old woman visited our hospital with a growing lump on her right breast accompanied by pain. Clinical and radiological examinations revealed a well-circumscribed 40-mm-diameter tumor at the inner lower quadrant of her right breast. Histological examination of a needle biopsy specimen revealed angiosarcoma. Based on a precise evaluation of the tumor by contrast-enhanced MRI and contrast-enhanced CT scan, a wide local excision with sufficient margins was performed. In the resected specimen, three discontinuous small lesions of angiosarcoma were observed around the main tumor. Therefore, total mastectomy was additionally performed. Pathological examination revealed two other small nodules of angiosarcoma in the remnant right breast, which appeared to be close but not continuous to the defective part of the initial resection. Postoperative follow-up at 1 year showed no signs of recurrence or distant metastasis. Multifocal primary breast angiosarcoma is extremely rare with only two previous reports describing its multifocality. CONCLUSIONS: Owing to its rarity, a standardized surgical treatment for breast angiosarcoma remains controversial. Our case suggests that primary breast angiosarcoma may occasionally present with multifocal tumor. Thus, it is important to keep in mind the multifocality of breast angiosarcoma when assessing its spread by diagnostic imaging and when planning the surgical strategy.

12.
Elife ; 72018 08 10.
Artigo em Inglês | MEDLINE | ID: mdl-30095406

RESUMO

The ultimate overexpression of a protein could cause growth defects, which are known as the protein burden. However, the expression limit at which the protein-burden effect is triggered is still unclear. To estimate this limit, we systematically measured the overexpression limits of glycolytic proteins in Saccharomyces cerevisiae. The limits of some glycolytic proteins were up to 15% of the total cellular protein. These limits were independent of the proteins' catalytic activities, a finding that was supported by an in silico analysis. Some proteins had low expression limits that were explained by their localization and metabolic perturbations. The codon usage should be highly optimized to trigger the protein-burden effect, even under strong transcriptional induction. The S-S-bond-connected aggregation mediated by the cysteine residues of a protein might affect its expression limit. Theoretically, only non-harmful proteins could be expressed up to the protein-burden limit. Therefore, we established a framework to distinguish proteins that are harmful and non-harmful upon overexpression.


Assuntos
Biologia Computacional/métodos , Dissulfetos/química , Regulação Fúngica da Expressão Gênica , Glicólise , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/metabolismo , Sequência de Aminoácidos , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/crescimento & desenvolvimento , Proteínas de Saccharomyces cerevisiae/química , Proteínas de Saccharomyces cerevisiae/genética , Homologia de Sequência
13.
Gastroenterol Res Pract ; 2018: 1024074, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29977284

RESUMO

OBJECTIVE: The serum pepsinogen test has limitation in its predictive power as a noninvasive biomarker for gastric cancer screening. We aimed to investigate whether the combination of TFF3 and pepsinogen could be an effective biomarker for the detection of gastric cancer even in the early stages. METHODS: In total, 281 patients with early gastric cancer (EGC), who underwent endoscopic submucosal dissection in Korea, and 708 healthy individuals from Japan were enrolled in the derivation cohort. The validation cohort included 30 Korean patients with EGC and 30 Korean healthy control blood donors. Serum TFF3 levels were examined using enzyme-linked immunosorbent assay. RESULTS: Using a cutoff of 6.73 ng/mL in the derivation cohort, the sensitivity of the combination of tests for EGC detection was superior (87.5%) to that of TFF3 (80.4%) or pepsinogen test alone (39.5%). Similarly, in the validation cohort, the sensitivity of TFF3 plus pepsinogen was higher (90.4%) than that of TFF3 (80.0%) or pepsinogen test alone (33.3%). CONCLUSION: The combination of serum TFF3 and pepsinogen is a more effective noninvasive biomarker for gastric cancer detection compared with pepsinogen or TFF3 alone, even in EGC. This trial is registered with NCT03046745.

14.
Sci Rep ; 7(1): 4846, 2017 07 07.
Artigo em Inglês | MEDLINE | ID: mdl-28687783

RESUMO

Breast cancer remains a common malignancy in women, but the take-up for breast cancer screening programs in Japan is still low, possibly due to its perceived inconvenience. TFF1 and TFF3 are expressed in both breast cancer tissue and normal breast. Serum trefoil proteins were reported as cancer screening markers for gastric, prostate, lung, pancreatic cancer and cholangio carcinoma. The purpose of this study was to examine whether serum trefoil proteins could be screening biomarkers for breast cancer. Serum trefoil proteins in 94 breast cancer patients and 84 health check females were measured by ELISA. Serum TFF1 and TFF3 were significantly higher and serum TFF2 was significantly lower in breast cancer patients. Area under the curve of receiver operating characteristic of TFF1, TFF2, and TFF3 was 0.69, 0.83, and. 0.72, respectively. AUC of the combination of TFF1, TFF2, and TFF3 was 0.96. Immunohistochemically, TFF1 expression was positive in 56.5% and TFF3 was positive in 73.9% of breast cancers, while TFF2 was negative in all tumors. Serum TFF1 had positive correlation with expression of TFF1 in breast cancer tissue. Serum concentrations of TFF1 and TFF3 but not TFF2 are higher in women with breast cancer than in women without breast cancer.


Assuntos
Biomarcadores Tumorais/sangue , Neoplasias da Mama/patologia , Soro/química , Fator Trefoil-1/sangue , Fator Trefoil-2/sangue , Fator Trefoil-3/sangue , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imuno-Histoquímica , Japão
15.
Genes Cells ; 22(6): 591-601, 2017 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-28503907

RESUMO

In budding yeast, a mother cell can produce a finite number of daughter cells over its life. The accumulation of a variety of types of damaged components has an impact on the aging process. Asymmetrical inheritance during cell division causes these aberrant intracellular constituents to be retained in mother cells and prevents them from segregating to daughter cells. However, the understanding of asymmetrical inheritance of individual proteins that are damaged or old age, and their relevance to the aging process, has been limited. The aim of this study is to propose a proteomics strategy for asymmetrical inheritance of preexisting proteins between mother and daughter cells. During synchronous culture for one generation, newly synthesized proteins were labeled with stable isotope amino acids to discriminate preexisting proteins originally expressed in mother cells, followed by separation of mother and daughter cells using a conventional method based on biotin labeling. Isotope incorporation ratios for individual proteins were quantified using mass spectrometry. We successfully identified 21 proteins whose preexisting versions were asymmetrically inherited in mother cells, including plasma membrane transporter involved in the aging process and organelle-anchoring proteins related to the stress response to misfolded proteins. Thus, our approach would be useful for making catalog of asymmetrically inherited proteins.


Assuntos
Proteoma/análise , Proteômica/métodos , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/citologia , Saccharomyces cerevisiae/metabolismo , Divisão Celular Assimétrica , Cromatografia Líquida , Saccharomyces cerevisiae/crescimento & desenvolvimento , Espectrometria de Massas em Tandem , Fatores de Tempo
16.
Plant Cell Physiol ; 57(11): 2312-2322, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27565204

RESUMO

Pattern recognition receptors on the plant cell surface mediate the recognition of microbe/damage-associated molecular patterns (MAMPs/DAMPs) and activate downstream immune signaling. Autophosphorylation of signaling receptor-like kinases is a critical event for the activation of downstream responses but the function of each phosphorylation site in the regulation of immune signaling is not well understood. In this study, 41 Ser/Thr/Tyr and 15 Ser/Thr residues were identified as in vitro and in vivo autophosphorylation sites of Arabidopsis CERK1, which is essential for chitin signaling. Comprehensive analysis of transgenic plants expressing mutated CERK1 genes for each phosphorylation site in the cerk1-2 background indicated that the phosphorylation of T479 in the activation segment and Y428 located upstream of the catalytic loop is important for the activation of chitin-triggered defense responses. Contribution of the phosphorylation of T573 to the chitin responses was also suggested. In vitro evaluation of kinase activities of mutated kinase domains indicated that the phosphorylation of T479 and T573 is directly involved in the regulation of kinase activity of CERK1 but the phosphorylation of Y428 regulates chitin signaling independently of the regulation of kinase activity. These results indicated that the phosphorylation of specific residues in the kinase domain contributes to the regulation of downstream signaling either through the regulation of kinase activity or the different mechanisms, e.g. regulation of protein-protein interactions.


Assuntos
Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimologia , Arabidopsis/imunologia , Quitina/farmacologia , Proteínas Serina-Treonina Quinases/química , Proteínas Serina-Treonina Quinases/metabolismo , Transdução de Sinais , Treonina/metabolismo , Tirosina/metabolismo , Sequência de Aminoácidos , Mutação , Fosforilação/efeitos dos fármacos , Imunidade Vegetal/efeitos dos fármacos , Plantas Geneticamente Modificadas , Domínios Proteicos , Transdução de Sinais/efeitos dos fármacos
17.
Proteomics ; 16(10): 1457-73, 2016 05.
Artigo em Inglês | MEDLINE | ID: mdl-27030420

RESUMO

The accurate and precise absolute abundance of proteins can be determined using mass spectrometry by spiking the sample with stable isotope-labeled standards. In this study, we developed a strategy of hierarchical use of peptide-concatenated standards (PCSs) to quantify more proteins over a wider dynamic range. Multiple primary PCSs were used for quantification of many target proteins. Unique "ID-tag peptides" were introduced into individual primary PCSs, allowing us to monitor the exact amounts of individual PCSs using a "secondary PCS" in which all "ID-tag peptides" were concatenated. Furthermore, we varied the copy number of the "ID-tag peptide" in each PCS according to a range of expression levels of target proteins. This strategy accomplished absolute quantification over a wider range than that of the measured ratios. The quantified abundance of budding yeast proteins showed a high reproducibility for replicate analyses and similar copy numbers per cell for ribosomal proteins, demonstrating the accuracy and precision of this strategy. A comparison with the absolute abundance of transcripts clearly indicated different post-transcriptional regulation of expression for specific functional groups. Thus, the approach presented here is a faithful method for the absolute quantification of proteomes and provides insights into biological mechanisms, including the regulation of expressed protein abundance.


Assuntos
Fragmentos de Peptídeos/química , Proteoma/química , Proteômica/normas , Proteínas de Saccharomyces cerevisiae/química , Escherichia coli , Estudos de Avaliação como Assunto , Fragmentos de Peptídeos/isolamento & purificação , Proteólise , Proteoma/isolamento & purificação , Padrões de Referência , Saccharomyces cerevisiae , Proteínas de Saccharomyces cerevisiae/isolamento & purificação , Espectrometria de Massas em Tandem , Tripsina/química
18.
Mol Cell Proteomics ; 15(1): 218-35, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26560065

RESUMO

Omics analysis is a versatile approach for understanding the conservation and diversity of molecular systems across multiple taxa. In this study, we compared the proteome expression profiles of four yeast species (Saccharomyces cerevisiae, Saccharomyces mikatae, Kluyveromyces waltii, and Kluyveromyces lactis) grown on glucose- or glycerol-containing media. Conserved expression changes across all species were observed only for a small proportion of all proteins differentially expressed between the two growth conditions. Two Kluyveromyces species, both of which exhibited a high growth rate on glycerol, a nonfermentative carbon source, showed distinct species-specific expression profiles. In K. waltii grown on glycerol, proteins involved in the glyoxylate cycle and gluconeogenesis were expressed in high abundance. In K. lactis grown on glycerol, the expression of glycolytic and ethanol metabolic enzymes was unexpectedly low, whereas proteins involved in cytoplasmic translation, including ribosomal proteins and elongation factors, were highly expressed. These marked differences in the types of predominantly expressed proteins suggest that K. lactis optimizes the balance of proteome resource allocation between metabolism and protein synthesis giving priority to cellular growth. In S. cerevisiae, about 450 duplicate gene pairs were retained after whole-genome duplication. Intriguingly, we found that in the case of duplicates with conserved sequences, the total abundance of proteins encoded by a duplicate pair in S. cerevisiae was similar to that of protein encoded by nonduplicated ortholog in Kluyveromyces yeast. Given the frequency of haploinsufficiency, this observation suggests that conserved duplicate genes, even though minor cases of retained duplicates, do not exhibit a dosage effect in yeast, except for ribosomal proteins. Thus, comparative proteomic analyses across multiple species may reveal not only species-specific characteristics of metabolic processes under nonoptimal culture conditions but also provide valuable insights into intriguing biological principles, including the balance of proteome resource allocation and the role of gene duplication in evolutionary history.


Assuntos
Proteínas Fúngicas/metabolismo , Kluyveromyces/metabolismo , Proteoma/metabolismo , Proteômica/métodos , Saccharomyces cerevisiae/metabolismo , Saccharomyces/metabolismo , Evolução Molecular , Proteínas Fúngicas/genética , Duplicação Gênica , Glucose/metabolismo , Glicerol/metabolismo , Kluyveromyces/genética , Kluyveromyces/crescimento & desenvolvimento , Proteoma/genética , Saccharomyces/genética , Saccharomyces/crescimento & desenvolvimento , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/crescimento & desenvolvimento , Especificidade da Espécie , Espectrometria de Massas em Tandem
19.
J Oleo Sci ; 61(11): 659-64, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-23138255

RESUMO

Cellobiose lipids (CLs) are bolaform glycolipid biosurfactants, which are produced from natural resources by a yeast strain and show fungicidal activity. In this study, the gelation properties of CL in solvents were investigated by several techniques including rheology and atomic force microscopy (AFM). The yeast CL was found to gelate 6 out of 26 solvents. Although it did not provide gels in ethanol or 1, 3-butanediol which are widely used for cosmetic industries, we succeeded in producing gels by mixing ethanol or 1, 3-butanediol with water. AFM observation of the gels on a silicon substrate provided 3D supramolecular structures with an entangled fibrous network. Moreover, it was also found that some of fibrous structures were twisted helical ribbons. This should be due to the cellobiose backbone having several chiral functional groups. The sol-gel phase transition temperatures for gels in mixed ethanol/water and 1, 3-butanediol/water systems were below 100°C, indicating that the gels can be obtained with rather mild preparation conditions. The present CL gels would be useful for novel multifunctional materials applicable to various industries.


Assuntos
Celobiose/química , Cryptococcus/química , Géis/química , Lipídeos/química , Tensoativos/química , Microscopia de Força Atômica , Peso Molecular , Reologia , Solventes/química , Temperatura , Fatores de Tempo
20.
Biosci Biotechnol Biochem ; 75(8): 1597-9, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21821939

RESUMO

Cryptococcus humicola JCM 1461 efficiently produced cellobiose lipids (CLs), bolaform biosurfactants. The main product was identified as 16-O-(2″,3″,4″,6'-tetra-O-acetyl-ß-cellobiosyl)-2-hydroxyhexadecanoic acid. The production yield of CLs reached 13.1 g/L under the intermittent feeding of glucose. The critical micelle concentrations (CMC) of the main product at pH 4.0 and 7.0 were 3.3×10(-5) M and 4.1×10(-4) M respectively.


Assuntos
Biotecnologia/métodos , Celobiose/metabolismo , Cryptococcus/metabolismo , Glicolipídeos/biossíntese , Ácidos Palmíticos/metabolismo , Tensoativos/metabolismo , Cromatografia Líquida de Alta Pressão , Cromatografia em Camada Fina , Meios de Cultura/química , Glucose/metabolismo , Concentração de Íons de Hidrogênio , Espectroscopia de Ressonância Magnética , Micelas , Ácidos Palmíticos/química , Propriedades de Superfície
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