CreA-independent carbon catabolite repression of cellulase genes by trimeric G-protein and protein kinase A in Aspergillus nidulans.

Cellulase production in filamentous fungi is repressed by various carbon sources. In our preliminary survey in Aspergillus nidulans, degree of de-repression differed depending on carbon sources in a mutant of creA, encoding the transcriptional repressor for carbon catabolite repression (CCR). To further understand mechanisms of CCR of cellulase production, we compared the effects of creA deletion with deletion of protein kinase A (pkaA) and G (ganB) genes, which constitute a nutrient sensing and signaling pathway. In plate culture with carboxymethyl cellulose and D-glucose, deletion of pkaA and ganB, but not creA, led to significant de-repression of cellulase production. In submerged culture with cellobiose and D-glucose or 2-deoxyglucose, both creA or pkaA single deletion led to partial de-repression of cellulase genes with the highest level by their double deletion, while ganB deletion caused de-repression comparable to that of the creA/pkaA double deletion. With ball-milled cellulose and D-glucose, partial de-repression was detected by deletion of creA but not of pkaA or ganB. The creA/pkaA or creA/ganB double deletion led to earlier expression than the creA deletion. Furthermore, the effect of each deletion with D-xylose or L-arabinose as the repressing carbon source was significantly different from that with D-glucose, D-fructose, and D-mannose. Consequently, this study revealed that PkaA and GanB participate in CreA-independent CCR and that contribution of CreA, PkaA, and GanB in CCR differs depending on the inducers, repressing carbon sources, and culture conditions (plate or submerged). Further study of CreA-independent mechanisms is needed to fully understand CCR in filamentous fungi.

Yeast Interspecies Comparative Proteomics Reveals Divergence in Expression Profiles and Provides Insights into Proteome Resource Allocation and Evolutionary Roles of Gene Duplication.

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.

Esthetic result of rhomboid flap repair after breast-conserving surgery for lower quadrant breast cancer lesion with skin invasion: report of two cases.

Breast-conserving surgery (BCS) has been increasingly performed as a standard operative strategy for patients with breast cancer. The primary purpose of BCS is to acquire both local control and good cosmetic results. An insignificant difference in cancer treatment results has been shown between BCS and total mastectomy. However, achieving sufficiently cosmetic results can be difficult, particularly in patients with tumors that are large or localized to the lower quadrant. To avoid breast deformities and asymmetries after BCS, immediate reconstruction using autologous tissue has been accepted as the standard option. Rhomboid skin and adipose flap repair is a simple, less invasive procedure than the myocutaneous flap, which has primarily been performed in patients with upper quadrant lesions. We herein report the cases of two patients with lower quadrant breast cancer with skin invasion, who underwent BCS with immediate breast repair using a rhomboid flap. This procedure is therefore worth considering as one of the first options for immediate repair after BCS.

[Clinical effect and positioning of capecitabine for metastatic breast carcinoma].

We evaluated the effect of capecitabine for metastatic breast carcinoma. Of 18 metastatic breast carcinoma patients experienced in our institution from November 2002 to July 2007, all patients had resisted the anthracycline or taxanes therapy before. Of these patients, 7 had liver, 6 had bone, and 5 had lung metastases. The capecitabine response in these 18 patients was evaluated as follows: PR in 4, long SD in 6, SD in 3, and PD in 5. In particular, 3 patients with liver metastases showed remarkable tumor regression. Compared to the 3rd-and 4th-line, 2nd-line capecitabine proved more effective. Hand-foot syndrome was found in 4 patients. Only one patient discontinued the therapy due to nausea. These results showed that capecitabine may be a useful treatment regimen for chemotherapy-resistant metastatic breast carcinoma patients.

Evaluation of lymph node metastases of breast cancer using ultrasmall superparamagnetic iron oxide-enhanced magnetic resonance imaging.

BACKGROUND: We assessed the utility of enhanced magnetic resonance imaging (MRI) using ultrasmall superparamagnetic iron oxide (USPIO) in the evaluation of axillary lymph node metastases in patients with breast cancer. STUDY DESIGN: MR examination of the axilla was performed before and 24-36 h after USPIO administration for patients with stage II or III breast cancer. Diagnostic performance was compared using size criteria (metastasis was defined when short axis diameter >5 or >10mm) or morphologic criteria on conventional MRI, the combined study of USPIO precontrast and postcontrast images, and USPIO postcontrast study alone. RESULTS: A total of 622 nodes (503 metastatic and 119 nonmetastatic nodes) were dissected from 33 patients. The results of conventional MRI for nodes >5mm were 59.1% sensitivity, 86.7% specificity, and 80.4% overall accuracy. Results for nodes >10mm were 15.7% sensitivity, 99.2% specificity, and 80.2% overall accuracy. Results based on morphology were 36.5% sensitivity, 94.1% specificity, and 81.0% overall accuracy. The results of the combined study of USPIO precontrast and postcontrast images were 86.4% sensitivity, 97.5% specificity, 91.1% positive predictive value, 96.1% negative predictive value, and 95.0% overall accuracy. The results of USPIO postcontrast images alone were 84.7% sensitivity, 96.8% specificity, and 94.0% overall accuracy. Patient-based results of postcontrast USPIO study alone were 100.0% sensitivity, 80.0% specificity, and 93.9% overall accuracy. CONCLUSIONS: USPIO postcontrast study alone was useful in the assessment of axillary lymph node metastases in patients with breast cancer.

Detection of human epidermal growth factor receptor 2 protein and gene in fine needle aspiration cytology specimens and tissue sections from invasive breast cancer: can cytology specimens take the place of tissue sections?

Overexpression of HER2 protein and HER2 gene amplification in breast cancer are prognostic factors for the response to specific medical treatments such as trastuzumab, endocrine therapy, and chemotherapy. Whereas HER2 expression and gene amplification are generally examined in tissue sections, we investigated whether specimens from fine needle aspiration cytology (FNAC) are adequate for these analyses. HER2 protein overexpression and HER2 gene amplification were assessed in both FNAC specimens and tissue sections from 58 cases of invasive breast cancer. Immunohistochemistry assay for HER2 protein expression was performed according to the HercepTest protocol, and HER2 gene amplification was examined with the Spot-light CISH (chromogenic in situ hybridization) Detection kit. There was a significant positive correlation between assessments of HER2 protein status in the cytology specimens and tissue sections. The sensitivity, specificity, and accuracy of HER2 gene amplification detection in cytology specimens in relation to those in tissue sections were 84.0% (21/25 cases), 87.9% (29/33 cases), and 86.2% (50/58 cases), respectively. FNAC specimens are suitable for detection of HER2 overexpression and HER2 gene amplification in invasive breast cancer.

[A case of liver metastasis of breast cancer successfully treated with paclitaxel infusion into the hepatic artery: an attempt of once weekly regimen].

On October 21, 1995, a 45-year-old woman underwent mastectomy (Kodama's procedure), preserving the thoracic muscles, based on the diagnosis of right breast cancer (T2aN1bM0). After surgery, she received 3 cycles of adjuvant chemotherapy (CAF) using anthracycline and other drugs. Thereafter she was orally treated with UFT for 2 years. She developed tumor recurrence in her liver 3 years after surgery. At that time, she was initially scheduled to undergo partial hepatectomy (including the tumor-affected area), but it was later judged that radical surgery was impossible due to the presence of multiple lesions in both lobes of the liver. A reservoir was therefore installed in the hepatic artery. Ten days after reservoir installation, intra-arterial infusion of paclitaxel (80 mg/m2), dissolved in 100 ml of physiological saline, was performed for one hour, using the installed reservoir. This treatment was administered once weekly for 3 consecutive weeks, followed by one week of rest. After 3 cycles of this regimen, the liver metastases had disappeared completely. At present, 2 years after the beginning of this therapy, the patient remains in a state of CR (complete response). Grade 2 alopecia and grade 1 peripheral neuropathy were the only adverse reactions observed.

[Examination of therapy using trastuzumab in patients with metastatic breast carcinoma].

We evaluated the effect of combination therapy of trastuzumab and paclitaxel for metastatic breast carcinoma. Among the 23 metastatic breast carcinoma patients treated in our institution from September 2001 to December 2002, 10 (43%) patients were immunohistochemically positive for the HER2 protein. Four of these patients had bone, 3 had lung, 2 had liver and 1 had supraclavicular lymph node metastases. The combination chemotherapy for these 10 patients was evaluated as follows: CR in 1 patient, PR in 4, NC in 2, and PD in 3. Two patients with liver metastases showed remarkable tumor regression. Combination therapy did not have to be stopped in any of the patients due to side effects. These results show that combination therapy of trastuzumab and paclitaxel may be a useful treatment regimen for chemotherapy-resistant metastatic breast carcinoma patients.

Intraductal lipid-rich carcinoma of the breast with a component of glycogen-rich carcinoma.

We report a rare case of intraductal lipid-rich carcinoma of the breast with a component of glycogen-rich carcinoma. An impalpable tumor that was revealed by mammography and magnetic resonance imaging was excised. Histologic examination showed vacuolated neoplastic cells in the mammary ducts, and electron microscopy confirmed lipid droplets in the cytoplasm. The coexistence of glycogen-rich carcinoma was shown. Lipid-rich carcinoma that is coexistent with glycogen-rich carcinoma is rare, and most lipid-rich carcinomas are invasive. Intraductal lipid-rich carcinoma is difficult to detect without echography or mammography.

High-resolution MR lymphography using ultrasmall superparamagnetic iron oxide (USPIO) in the evaluation of axillary lymph nodes in patients with early stage breast cancer: preliminary results.

BACKGROUND: The purpose of this study was to examine the utility of high-resolution magnetic resonance (MR) lymphography using ultrasmall superparamagnetic iron oxide (USPIO) in the evaluation of axillary lymph nodes in patients with early stage breast cancer. METHODS: Ten women with breast cancer without swollen axillary lymph nodes were enrolled in this study. High-resolution MR lymphography was performed 24 h after administration of USPIO. On the MR examination, a 3-inch surface coil was placed on the sentinel lymph node (SLN) parts as defined by a radioisotope (RI) scintigraphy method, and T2*-weighted (T2*W) and T1-weighted (T1W) images were obtained. Detected nodes were differentiated as normal or diseased nodes by the enhancement patterns. The day after MR examination, SLN biopsy (SNB) was performed. The imaging results were compared to the histopathologic findings. RESULTS: On MR images, the mean number of detectable nodes was 7.9 per patient. Eight patients who histopathologically had no metastatic lymph nodes were diagnosed as nonmetastatic and two patients who had 3- and 6-mm metastatic areas in the node, respectively, were diagnosed as metastatic preoperatively. No side effects were noted. CONCLUSIONS: High-resolution MR lymphography using USPIO enabled us to obtain good axillary lymph node evaluation results. These results suggest that this method of imaging may contribute to better preoperative lymph node staging.

Sentinel lymph node biopsy in breast cancer patients undergoing skin/nipple-sparing mastectomy and immediate autologous reconstruction.

Immediate breast reconstruction following mastectomy is an effective treatment for breast cancer patients. Among several implant options, a latissimus dorsi myocutaneous (LDM) flap is used mainly due to the ease and minimal invasiveness of the procedure. The role of sentinel lymph node (SLN) biopsy with total mastectomy is evolving. Since SLN biopsy is not included in health insurance coverage in the treatment of patients in Japan, it is not generally performed as a separate procedure due to its cost. The present study reviewed the results of seven patients who underwent initial-staged SLN biopsy followed by planned mastectomy and LDM flap reconstruction. Two patients with positive SLNs showed macrometastases and underwent modified radical mastectomy with immediate reconstruction. In contrast, cases showing negative results for sentinel lymph nodes underwent total mastectomy. There were no false-negative cases among the SLN biopsy-negative cases. When an SLN is found to be positive on final pathology, the patient with reconstruction by LDM flap generally requires a potentially difficult reoperation on the remaining axillary nodes. When initial SLN biopsy is generally performed as a separate procedure in Japan, it will be an effective method for screening the axilla for patients who wish to undergo LDM flap reconstruction.

Examination of the clinical usefulness of TS-1 in locally recurrent breast cancer.

This study examined the therapeutic efficacy of TS-1 in the treatment of locally recurrent breast cancer. Between August 2006 and May 2009, 7 patients with local breast cancer recurrences were selected included in the study. The sites of recurrence were the cervical lymph nodes in 4 patients, the axillary lymph nodes in 2 and the thoracic wall in 1. Among the 7 patients, 6 were administered TS-1 as first-line treatment and 1 was administered TS-1 as second-line treatment. Complete response (CR) was achieved in 2 patients, 1 achieved partial response, 2 had stable disease (SD), 1 had long SD and 1 had progressive disease. The overall response rate was 43%, and the clinical benefit rate was 57%. A patient with recurrence of breast cancer in the thoracic wall 28 years after surgery also achieved CR following therapy. The only adverse event observed was a case of hand-foot syndrome in 1 patient. No patients withdrew from treatment, and favorable compliance was achieved in the study. The results indicated that TS-1 has the potential to be one of the drugs for first-line treatment of locally recurrent breast cancer.

Usefulness of intraoperative touch smear cytology in breast-conserving surgery.

In breast-conserving surgery, positive margins are closely related to intramammary recurrence, but methods of assessing resection stumps during breast-conserving surgery have not been standardized. The present study investigated the usefulness of intraoperative touch smear cytology in our department. From 2005 to 2008, a total of 420 patients underwent breast cancer surgery. Subjects comprised 160 patients who underwent breast-conserving surgery and touch smear cytology. Results of the touch smear cytology were compared to those of the histological tissue analysis. Touch smear cytology displayed 70% sensitivity (14/20), 97.1% specificity (136/140) and a diagnostic accuracy of 93.8% (150/160). Six false-negative cases and 4 false-positive cases were identified. Of the 6 false-negatives, cancer cells were noted in the ductal component in 5 cases, and the degree of cancer cell atypia in the stump was low. Residual cancer cells were noted in the stump in 18 cases, and additional resection was performed in 16 cases. Cancer cells were identified histologically in the additionally resected tissue in 8 of these 16 cases (50%). The direction of positive cytology was towards the nipple in 16 cases, lateral tissue in 5 cases and contralateral nipple in 2 cases. A greater cancer cell volume, as assessed by touch smear cytology, tended to be associated with higher frequency of positive margins, as assessed by histological tissue analysis. Touch smear cytology is easy to perform, offering a very useful technique yielding comparable results to histological tissue analysis.

Tau expression and efficacy of paclitaxel treatment in metastatic breast cancer.

PURPOSE: Paclitaxel is widely used for the treatment of patients with metastatic breast cancer (MBC). Although several mechanisms of paclitaxel resistance have been demonstrated, useful markers of paclitaxel resistance have not been available in clinical practice. METHODS: In this study, the clinical significance of tau expression in MBC cases was established by identifying candidates with paclitaxel administration. Tissue specimens obtained from 35 patients were examined. Status of tau expression was determined by immunohistochemistry. RESULTS: Fifteen cases were classified as tau-negative and 20 cases were classified as tau-positive, respectively. Sixty percent of tau-negative expression showed favorable response. Conversely, 85% of tau-positive expression showed progressive or stable disease after paclitaxel administration. Time to disease progression in tau-negative and tau-positive groups was 9.4 +/- 6.6 and 6.0 +/- 3.7 months, respectively. CONCLUSIONS: Patients with tau-positive expression may derive less benefit than tau-negative from paclitaxel therapy in MBC.

Prognostic value of tumor-infiltrating dendritic cells expressing CD83 in human breast carcinomas.

DCs are the most potent antigen-presenting cells that play a major role in initiating the antitumor immune response. Although the clinical significance of TIDCs has been investigated in a variety of human cancers, few studies have focused on the in situ maturation status of DCs. We have analyzed the maturation-specific significance of TIDCs in the prognosis of patients with breast carcinoma. We evaluated 130 breast carcinomas for the presence of TIDCs using immunohistochemistry with an anti-CD1a antibody for immature DCs and an anti-CD83 antibody for mature DCs. Intratumoral expression of immunosuppressive cytokines was also examined. All samples contained CD1a(+) TIDCs, and 82 (63.1%) samples contained CD83(+) TIDCs. The number of CD83(+) TIDCs was inversely correlated with lymph node metastasis and with tissue expression of VEGF and TGF-beta, whereas the number of CD1a(+) TIDCs was not. Kaplan-Meier analysis (log rank statistics) revealed a significant association of increasing number of CD83(+) TIDCs with longer relapse-free (p = 0.002) and overall (p < 0.001) survival. Furthermore, among patients with lymph node metastasis, the survival rate of those with larger numbers of CD83(+) TIDCs was significantly better than that of patients with fewer CD83(+) TIDCs. Multivariate analysis revealed that CD83(+) TIDCs had independent prognostic relevance in breast carcinomas. The infiltration of tumors by mature DCs expressing CD83 may be of great importance in initiating the primary antitumor immune response and is confirmed as an independent, immunologic prognostic parameter for survival in patients with breast cancer.