Stress-dependent cell stiffening by tardigrade tolerance proteins that reversibly form a filamentous network and gel.

Tardigrades are able to tolerate almost complete dehydration by entering a reversible ametabolic state called anhydrobiosis and resume their animation upon rehydration. Dehydrated tardigrades are exceptionally stable and withstand various physical extremes. Although trehalose and late embryogenesis abundant (LEA) proteins have been extensively studied as potent protectants against dehydration in other anhydrobiotic organisms, tardigrades produce high amounts of tardigrade-unique protective proteins. Cytoplasmic-abundant heat-soluble (CAHS) proteins are uniquely invented in the lineage of eutardigrades, a major class of the phylum Tardigrada and are essential for their anhydrobiotic survival. However, the precise mechanisms of their action in this protective role are not fully understood. In the present study, we first postulated the presence of tolerance proteins that form protective condensates via phase separation in a stress-dependent manner and searched for tardigrade proteins that reversibly form condensates upon dehydration-like stress. Through a comprehensive search using a desolvating agent, trifluoroethanol (TFE), we identified 336 proteins, collectively dubbed "TFE-Dependent ReversiblY condensing Proteins (T-DRYPs)." Unexpectedly, we rediscovered CAHS proteins as highly enriched in T-DRYPs, 3 of which were major components of T-DRYPs. We revealed that these CAHS proteins reversibly polymerize into many cytoskeleton-like filaments depending on hyperosmotic stress in cultured cells and undergo reversible gel-transition in vitro. Furthermore, CAHS proteins increased cell stiffness in a hyperosmotic stress-dependent manner and counteract the cell shrinkage caused by osmotic pressure, and even improved the survival against hyperosmotic stress. The conserved putative helical C-terminal region is necessary and sufficient for filament formation by CAHS proteins, and mutations disrupting the secondary structure of this region impaired both the filament formation and the gel transition. On the basis of these results, we propose that CAHS proteins are novel cytoskeleton-like proteins that form filamentous networks and undergo gel-transition in a stress-dependent manner to provide on-demand physical stabilization of cell integrity against deformative forces during dehydration and could contribute to the exceptional physical stability in a dehydrated state.

[Primary breast diffuse large B-cell lymphoma in an elderly man maintaining long-term complete response].

Primary breast diffuse large B-cell lymphoma (DLBCL) is a rare non-Hodgkin's lymphoma that mostly affects women. Here, we report a case of primary breast DLBCL that affected an older man without any autoimmune disease or drug-related female hormones. The patient was a 65-year-old man whose chief complaints were gradually-increasing lump in the right chest and swelling of the right axillary lymph nodes. He was diagnosed with malignant lymphoma through a needle biopsy on suspicion of right breast cancer with right axillary lymph node metastasis. Since the histological type could not be confirmed, right breast mass resection was performed. The patient was referred to our department for treatment because of the diagnoses of primary breast DLBCL, germinal center B-cell type (Hans classification), and clinical stage IIA. In addition to the six courses of R-CHOP therapy, intrathecal injections were used in combination to prevent CNS infiltration. He has been in complete remission for 5 years. Although rare, breast lymphoma can also occur in men; therefore, early histological diagnosis and response to CNS recurrence prevention are important.

Rapid HER2 cytologic fluorescence in situ hybridization for breast cancer using noncontact alternating current electric field mixing.

BACKGROUND: Human epidermal growth factor receptor 2-in situ hybridization (HER2-ISH) is widely approved for diagnostic, prognostic biomarker testing of formalin-fixed paraffin-embedded tissue blocks. However, cytologic ISH analysis has a potential advantage in tumor samples such as pleural effusion and ascites that are difficult to obtain the histological specimens. Our aim was to evaluate the clinical reliability of a novel rapid cytologic HER2 fluorescence ISH protocol (rapid-CytoFISH). MATERIALS AND METHODS: Using a new device, we applied a high-voltage/frequency, noncontact alternating current electric field to tissue imprints and needle rinses, which mixed the probe within microdroplets as the voltage was switched on and off (AC mixing). Cytologic samples (n = 143) were collected from patients with immunohistochemically identified HER2 breast cancers. The specimens were then tested using standard dual-color ISH using formalin-fixed paraffin-embedded tissue (FFPE-tissue DISH) for HER2-targeted therapies, CytoFISH, and rapid-CytoFISH (completed within 4 h). RESULTS: All 143 collected cytologic specimens (50 imprinted cytology specimens from resected tumors and 93 liquid-based cytology specimens from needle rinses) were suitable for FISH analysis. The HER2/chromosome enumeration probe (CEP) 17 ratios did not significantly differ between FFPE-tissue DISH and either CytoFISH protocol. Based on HER2 scoring criteria, we found 95.1% agreement between FFPE-tissue DISH and CytoFISH (Cohen's kappa coefficient = 0.771 and 95% confidence interval (CI): 0.614-0.927). CONCLUSION: CytoFISH could potentially serve as a clinical tool for prompt determination of HER2 status in breast cancer cytology. Rapid-CytoFISH with AC mixing will enable cancer diagnoses and HER2 status to be determined on the same day a patient comes to a clinic or hospital.

Evaluation of the potential for lymph node metastasis using CRP 1846C>T genetic polymorphism in invasive breast cancer.

Lymph node status is a key indicator of the best approach to treatment of invasive breast cancer. However, the accuracy with which lymph node metastasis is diagnosed is not currently satisfactory. New and more reliable methods that enable one to know who has a greater potential for lymph node metastasis would be highly desirable. We previously reported that lymph node involvement in esophageal and lung cancer may have a genetic component: C-reactive protein (CRP) 1846C>T genetic polymorphism. Here we examined the diagnostic value of CRP 1846C>T polymorphism for assessing the risk of lymph node metastasis in cases of invasive breast cancer. The study participants were 185 women with invasive breast cancer who underwent curative surgery with lymph node dissection. Using DNA from blood samples and polymerase chain reaction-restriction fragment length polymorphism, the utility of CRP genetic 1846C>T polymorphism (rs1205) for assessing the risk of lymph node metastasis was evaluated. Fifty-two (28 %) patients had lymph node metastasis. After the patients were divided into two groups based on their CRP 1846 genotypes (C/C+C/T and T/T), the clinical characteristics did not differ between the groups, but there was a significantly greater incidence of lymph node metastasis among patients in the T/T group. Moreover, the odds ratio for lymph node involvement in patients carrying the 1846 T/T genotype was more than 2.2 in multivariate logistic regression models. CRP genetic polymorphism may be a novel predictor of the risk of lymph node metastasis in invasive breast cancer.

[A case of recurrent malignant melanoma of esophagus responsive to combined chemotherapy, radiotherapy and cellular immunotherapy].

Treatment of primary malignant melanoma of the esophagus remains challenging. We treated a 53-year-old man with pT4N2M0, Stage IVa malignant melanoma of the esophagus with esophagectomy followed by adjuvant chemotherapy. Six months later, computed tomography revealed a 12 cm disseminated tumor of the mesenterium, multiple peritoneal dissemination, and a large amount of ascites. We administered chemotherapy consisting of dacarbazine combined with cisplatin and nimustine, and radiotherapy(50 Gy)was applied to the disseminated mesenteric tumor. At another clinic, the patient was administered synchronous cellular immunotherapy consisting of dendritic cells pulsed with autologous tumor lysates and lymphokine-activated killer cells. The mesenteric tumor was extremely responsive to this trimodal treatment. Because recurrence occurred later within the left orbita muscle, we added 50 Gy of radiation to prevent blindness. The patient responded to this treatment and survived another 6 months with high quality of life. It is difficult to treat advanced malignant melanoma of the esophagus, and patient prognosis is extremely poor. In this patient, the recurrent tumors responded well to trimodal therapy consisting of chemotherapy, radiotherapy and cellular immunotherapy.

Pseudoangiomatous stromal hyperplasia (PASH) of the mammary gland: report of a case.

Pseudoangiomatous stromal hyperplasia (PASH) is an uncommon benign breast disease. Pseudoangiomatous stromal hyperplasia is characterized by a collagenous proliferation of mammary stroma, associated with capillary-like spaces. We herein report the case of a 48-year-old woman who presented with a huge mass in her right breast. Physical examination revealed a 20 x 23-cm mass, which was surgically excised. Histopathologically, the tumor was composed of abundant fibrous stroma containing non-vascular slit-like spaces and scattered ducts and lobules. Neither stromal nor epithelial cells showed atypia. We therefore diagnosed the tumor to be PASH.

[A case of peritoneal dissemination of postoperative primary duodenal cancer successfully treated by TS-1 therapy].

A 62-year-old man reported to our hospital with serious complaints of abdominal pain, vomiting, and weight loss. An endoscopic examination detected a type 2 tumor of the descending limb of the duodenum. With a diagnosis of adenocarcinoma based on the biopsy finding, the patient was subjected to surgery. Laparotomy revealed the presence of a duodenal tumor disseminating to the omentum at the site where the transverse colon is attached. Pancreatoduodenectomy and partial resection of the transverse colon were carried out. CT conducted 6 months after surgery did not show any signs of tumor recurrence; but one year later, extensive tumor dissemination was noted on the hepatic surface. Upon consultation with the patient, a regimen of 80 mg/day of TS-1 given for 4 weeks followed by 2 weeks of a drug-free period was initiated. Six months later, the growth of tumor became arrested, improving his QOL. Nine months later, the tumor growth was progressive and the patient died two years after operation. The patient could gain long-term survival after operation. The TS-1 regimen applied in the present case may constitute a therapeutic strategy to be considered for similar conditions in future.

[A case of effective weekly paclitaxel administration for gastric cancer recurrence with carcinomatous pericarditis].

We reported a case of effective weekly paclitaxel administration for gastric cancer recurrence with carcinomatous pericarditis. A 69-year-old man underwent distal gastrectomy for gastric cancer in December 2001. However, he was re-admitted to the hospital for dyspnea in November 2002. A diagnosis of cardiac tamponade caused by gastric cancer relapse was made, and the patient was treated by weekly paclitaxel administration (90 mg/body) after drainage. It was effective in preventing reaccumulation of the pericardial effusion until his death 73 days after the diagnosis. It is thought that weekly paclitaxel administration can be a treatment for carcinomatous pericarditis.

Use of autologous instead of allogeneic blood transfusion during esophagectomy prolongs disease-free survival among patients with recurrent esophageal cancer.

BACKGROUND AND OBJECTIVES: A substantial body of evidence suggests that allogeneic blood transfusion increases the rate of recurrence of resected malignancies. The present study was conducted with the aim of understanding better the clinical characteristics of recurrent esophageal cancer and determining whether any survival advantage is conferred by transfusing autologous instead of allogeneic blood during the esophagectomy for the original malignancy. METHODS: We retrospectively analyzed 123 patients who received blood transfusion while undergoing esophagectomy for thoracic esophageal cancer between January 1991 and February 1998. We focused on those patients in whom the malignancy recurred. Of them, 23 patients received allogeneic blood and 18 received autologous blood. Compared were the clinico-pathological factors influencing prognosis as well as the disease-free survival periods and the period of survival after recurrence of the cancer. RESULTS: The clinico-pathological factors that influenced prognosis were similar in the two groups. There was also no significant difference in the rate at which the esophageal cancer recurred, or in survival time once it had recurred. On the other hand, disease-free survival prior to recurrence was significantly prolonged in the autologous blood transfusion group. CONCLUSION: Use of autologous instead of allogeneic blood prolongs disease-free survival of esophageal cancer patients.

Survival advantage of using autologous blood transfusion during surgery for esophageal cancer.

PURPOSE: There is evidence that blood transfusion is associated with an increased rate of tumor recurrence. This study was conducted to assess the survival advantage of giving autologous blood instead of allogeneic blood during surgery for esophageal cancer. METHODS: We retrospectively analyzed 62 patients who underwent esophagectomy for thoracic esophageal cancer between January 1991 and February 1995 and received allogeneic blood transfusion, and 61 patients operated on between March 1995 and February 1998, who received autologous blood transfusion. The clinicopathological factors and survival rates were compared between the two groups. RESULTS: The clinicopathological factors that influenced prognosis were similar in the two groups; however, a definite survival advantage was evident in the autologous blood transfusion group. According to multivariate analyses, the transfusion of allogeneic blood was an independent prognostic factor ( P = 0.0222), as was the presence of metastatic lymph nodes. Patients who received allogeneic blood transfusions perioperatively had more than a twofold greater risk (Hazard ration 2.406) of death over patients who received autologous blood transfusions. CONCLUSION: Autologous blood transfusion appears to be an independent prognostic factor for the survival of patients with esophageal cancer.