Understanding the molecular mechanisms of odorant binding and activation of the human OR52 family.

Structural and mechanistic studies on human odorant receptors (ORs), key in olfactory signaling, are challenging because of their low surface expression in heterologous cells. The recent structure of OR51E2 bound to propionate provided molecular insight into odorant recognition, but the lack of an inactive OR structure limited understanding of the activation mechanism of ORs upon odorant binding. Here, we determined the cryo-electron microscopy structures of consensus OR52 (OR52cs), a representative of the OR52 family, in the ligand-free (apo) and octanoate-bound states. The apo structure of OR52cs reveals a large opening between transmembrane helices (TMs) 5 and 6. A comparison between the apo and active structures of OR52cs demonstrates the inward and outward movements of the extracellular and intracellular segments of TM6, respectively. These results, combined with molecular dynamics simulations and signaling assays, shed light on the molecular mechanisms of odorant binding and activation of the OR52 family.

Effectiveness of pegfilgrastim prophylaxis in preventing febrile neutropenia during R-FC chemoimmunotherapy for chronic lymphocytic leukemia: A multicenter prospective phase II study.

Background: A chemotherapy of rituximab, fludarabine and cyclophosphamide (R-FC) has been accepted as a promising frontline chemotherapy in selected patients with chronic lymphocytic leukemia (CLL). Although R-FC regimen is a relatively dose-dense regimen and neutropenia incidence is more than 50%, primary prophylactic pegfilgrastim was not fully recommended in the clinical field. Therefore, the study evaluated the prophylactic effectiveness of pegfilgrastim to reduce the incidence of febrile neutropenia associated with R-FC of patients with CLL. Patients and methods: A single-arm, multicenter, prospective phase II study was designed to assess the efficacy of prophylactic pegfilgrastim. Thirty-four CLL patients were enrolled and analyzed for neutropenia and other related factors, and comparative analysis was performed with historical cohort. Results: Compared with our historical cohort, incidence of grade 3-4 neutropenia and febrile neutropenia was remarkably reduced during any cycle of chemotherapy (14.7% vs. 48.2% of study cohort vs. historical cohort during C1, 5.9% vs. 65.8% during C2, 12.9% vs. 80.6% during C3, 10% vs. 84.6% during C4, 3.4% vs. 83.6% during C5, and 10.7% vs. 85.7% during C6, p <0.001). Also, cumulative incidence of disrupted chemotherapy was noticeably reduced in study cohort on any cycles of R-FC regimen (8.8% vs. 22.2% of study cohort vs. historical cohort on C2, 9.7% vs. 25.2% on C3, 13.4% vs. 26.9% on C4, 13.8% vs. 45.2% on C5, 17.9% vs. 47.3% on C6, p=0.007). In addition, treatment-related mortality was 5.9%, which significantly reduced compared to 9.6% of our historical cohort (HR 0.64, 95% CI 0.42-0.79, P = 0.032). Conclusion: Primary prophylactic pegfilgrastim is effective in the prevention of neutropenia/febrile neutropenia, and infection-related mortality during R-FC regimen in patients with CLL.

Structural basis for Y2 receptor-mediated neuropeptide Y and peptide YY signaling.

Neuropeptide Y (NPY) and its receptors are expressed in various human tissues including the brain where they regulate appetite and emotion. Upon NPY stimulation, the neuropeptide Y1 and Y2 receptors (Y1R and Y2R, respectively) activate GI signaling, but their physiological responses to food intake are different. In addition, deletion of the two N-terminal amino acids of peptide YY (PYY(3-36)), the endogenous form found in circulation, can stimulate Y2R but not Y1R, suggesting that Y1R and Y2R may have distinct ligand-binding modes. Here, we report the cryo-electron microscopy structures of the PYY(3-36)‒Y2R‒Gi and NPY‒Y2R‒Gi complexes. Using cell-based assays, molecular dynamics simulations, and structural analysis, we revealed the molecular basis of the exclusive binding of PYY(3-36) to Y2R. Furthermore, we demonstrated that Y2R favors G protein signaling over ß-arrestin signaling upon activation, whereas Y1R does not show a preference between these two pathways.

Peripheral T cell lymphoma of the nasopharynx with expansion of EBV-positive B cells masquerading as an extranodal NK/T cell lymphoma, nasal type.

Epstein-Barr virus-infected B cells are found at high frequency in peripheral T cell lymphoma. Herein, we report a case involving excessive EBV-positive B cells accompanying peripheral T cell lymphoma, not otherwise specified in the nasopharynx masquerading as nasopharyngeal extranodal NK/T cell lymphoma. A large number of Epstein-Barr virus-infected B cells infiltrate in between CD3-positive cytotoxic tumor T cells, as if EBV was infecting tumor T cells. After chemotherapy, the T cell lymphoma population decreased, but the B cell population expanded to form EBV-positive diffuse large B cell lymphoma in the tonsils and nasopharynx. At the follow-up, bone marrow biopsy exhibited infiltration of composite peripheral T cell lymphoma, not otherwise specified, and EBV-positive diffuse large B cell lymphoma. Although this condition is rare, the cell lineage of EBV-infected cells must be confirmed when diagnosing extranodal NK/T cell lymphoma to exclude the possibility of misdiagnosis by Epstein-Barr virus-infected B cells.

Composite follicular lymphoma and classic Hodgkin lymphoma.

Composite lymphoma is very rare and a combination of Hodgkin lymphoma and non-Hodgkin lymphoma and even histiocytic tumors can occur. Because of the unfamiliarity, not only can this cause diagnostic problems, but can also affect treatment plan. We report a case of composite lymphoma in a 40-year-old male. Initial biopsy showed a composite lymphoma of follicular lymphoma grade 1 and classic Hodgkin lymphoma. After chemotherapy, another lymph node was taken because of disease progression, which revealed follicular lymphoma, grade 3a without Hodgkin lymphoma component.

Treatment outcomes of stereotactic ablative radiation therapy for non-spinal bone metastases: focus on response assessment and treatment indication.

OBJECTIVE: To report treatment outcomes of stereotactic ablative radiation therapy (SABR) for non-spinal bone metastases in a single institution, and to compare assessments of Response Evaluation Criteria in Solid Tumors (RECIST) v. 1.1 and the University of Texas MD Anderson Cancer Center (MDA) criteria. METHODS: From July 2011 to January 2017, 33 patients with 38 non-spinal bone metastatic lesions were treated using SABR. Treatment intent was categorized as follows: single metastasis or oligo-metastases; oligo-progression; and dominant areas of progression. Tumor responses were evaluated according to the RECIST and MDA criteria. Local control (LC) was defined as lesions that were not classified as progressive disease on both criteria. RESULTS: The median follow-up period was 10.4 months (range, 2.5-47.4). Both 1- and 2 year LC rates were 94.2 %. The median overall survival (OS) was 20.2 months, and the median progression-free survival (PFS) was 6.9 months. Treatment intent was a significant factor for OS in multivariate analysis. The 1 year OS rates for single metastasis or oligo-metastasis, for oligo-progression, and for dominant areas of progression were 84.2%, 66.7%, and 0.0%, respectively ( p < 0.001). Overall response rate was 86.8 % according to MDA criteria, and 75.7 % according to RECIST criteria. When using MDA criteria, there appeared to be significant associations both between response and PFS (median 7.6 months for responders vs 2.5 months for non-responders; p = 0.036) and between response and OS. In contrast, when using RECIST criteria, the associations were significant neither between response and PFS (median 5.8 months for responders vs 9.3 months for non-responders; p = 0.522) nor between response and OS (25.7 months for responders vs 18.5 months for non-responders; p = 0.811). CONCLUSION: SABR for non-spinal bone metastases demonstrated high LC rates with acceptable toxicity. The MDA criteria demonstrated advantages in predicting survival outcomes. ADVANCES IN KNOWLEDGE: SABR for non-spinal bone metastases is a promising treatment option to achieve good local control. The MDA criteria, which is a newly proposed response evaluation criteria for bone metastases, has advantages in predicting survival outcomes compared to other established criteria.

Tripodal Lipoprotein Variants with C-Terminal Hydrophobic Residues Allosterically Modulate Activity of the DegP Protease.

DegP, a member of the highly conserved HtrA family of proteases, performs a regulated proteolysis of toxic misfolded proteins in the periplasm of Gram-negative bacteria. The allosteric switch between inactive and active conformations is a central mechanism to carefully control proteolytic activity of DegP and to maintain the optimal cellular fitness, but few other molecules than substrates are known to allosterically control DegP activity. Here, we demonstrate that a mutant variant of an outer-membrane lipoprotein, Lpp+Leu, can function as a novel allosteric effector that changes the dynamic range of DegP activity. The three leucines at the C-termini of trimeric Lpp+Leu are central components for activity modulation. Selection experiments with Lpp variant libraries show that Lpp variants with diverse sequences at or near C-termini, in particular those with hydrophobic residues at C-termini, function similarly to Lpp+Leu. Interestingly, Lpp variants carrying different residues at C-terminal, penultimate, or antepenultimate positions display dramatically different patterns of activation and inhibition effects, suggesting that their interactions with DegP differentially stabilize distinct inactive or active conformations. We propose that the tripodal structure with three hydrophobic ends that mimics Lpp+Leu is a novel platform for allosteric effectors, which may be useful in developing new antibiotics against DegP or homologous HtrA proteases.

Carbon ion irradiation abrogates Lin28B-induced X-ray resistance in melanoma cells.

The Lin28/let-7 axis plays an important role in tumor initiation and developmental processes. Lin28B is upregulated in a variety of cancers, and its overexpression enhances cancer cell proliferation and radioresistance through the suppression of let-7 micro RNA expression. In this study, we investigated the role of the Lin28/let7 axis as a target for radiosensitization of melanoma cancer cells. The overexpression of Lin28B reduced mature let-7 microRNA expression in melanoma cell lines, and enhanced the sphere-forming ability of melanoma cell lines, which is a characteristic of cancer stem cell (CSC) populations. Interestingly, Lin28B-overexpressed melanoma cells were more resistant to X-ray irradiation than control cells, and Lin28B-induced radioresistance was abolished after carbon ion irradiation. Consistent with these results, Lin28B overexpression reduced the numbers of γH2A.X foci after X-ray irradiation, whereas carbon ion irradiation had no such effect. Our results suggest that a carbon ion beam is more effective than an X-ray beam in terms of killing cancer cells, possibly due to elimination of CSC populations.

Stereotactic Body Radiotherapy for Recurrent or Oligometastatic Uterine Cervix Cancer: A Cooperative Study of the Korean Radiation Oncology Group (KROG 14-11).

AIM: To evaluate local control and patient survival for recurrent or oligometastatic uterine cervical cancer treated with stereotactic body radiotherapy (SBRT) using CyberKnife, and to demonstrate the safety of SBRT. PATIENTS AND METHODS: Between 2002 and 2013, 100 recurrent or oligometastatic lesions in 85 patients were treated with SBRT at three Institutions. SBRT sites were within the previous RT field in 59 and partially overlapped in nine. SBRT sites included three local recurrences, 89 lymph node metastases, and eight distant metastases. Patients were treated with a median dose of 39 Gy in three fractions, which was equivalent to a biologically effective dose (BED) of 90 Gy. RESULTS: The median follow-up period was 20.4 months. Local failure occurred in 17 out of 100 SBRT-treated sites. The 2-year and 5-year local progression-free survival rates were 82.5% and 78.8%, respectively. Eleven local failures occurred within the previous RT field. The 2-year and 5-year overall survival rates were 57.5% and 32.9%, respectively. BED >90 Gy (p=0.072) and >69 Gy (p=0.059) and longer disease-free interval (p=0.065) predicted marginally superior local control. Re-irradiation appeared to be related to inferior local control (p<0.001), but the SBRT BED in this group was much lower than the dose in the other group (median BED, 79 Gy vs. 90 Gy). Chronic toxicities of grade 3 or more occurred in five cases. CONCLUSION: SBRT for recurrent or oligometastatic cervical cancer resulted in excellent local control, especially with a long disease-free interval and high BED treatment, with acceptable toxicities. Therefore, SBRT can be considered a therapeutic option for these patients.

Robotic system-based fractionated stereotactic radiotherapy in locally recurrent nasopharyngeal carcinoma.

PURPOSE: We reviewed survival, local control, and toxicity in patients with locally recurrent nasopharyngeal carcinoma (NPC) who had been treated with fractionated stereotactic radiotherapy (FSRT). MATERIALS AND METHODS: Between June 2002 and March 2008, we retrospectively reviewed 35 patients with locally recurrent NPC treated using FSRT with CyberKnife. Gross tumor volumes ranged from 2.6 to 64.0 ml (median, 7.9 ml). Radiation doses were prescribed at the isodose line (75-84% of the maximum dose; median, 80%). The prescribed dose of FSRT ranged from 24 to 45 Gy (median, 33 Gy) in three or five fractions. RESULTS: The overall survival (OS) rate, local failure-free survival (LFFS) rate, and disease progression-free survival (DPFS) rate at 5 years were 60%, 79%, and 74%, respectively. Twenty-three patients achieved complete response after FSRT. Only T stage at recurrence was an independent prognostic factor for OS and DPFS. Five patients exhibited severe late toxicity (Grade 4 or 5). CONCLUSIONS: With regard to OS and LFFS, our study provided favorable outcomes. The incidence of severe late toxicities was acceptable in our study. FSRT would be considered as the alternative treatment of choice in re-irradiation for locally recurrent NPC.

Stereotactic body radiation therapy boost in locally advanced pancreatic cancer.

PURPOSE: To investigate the clinical application of a stereotactic body radiation therapy (SBRT) boost in locally advanced pancreatic cancer patients with a focus on local efficacy and toxicity. METHODS AND MATERIALS: We retrospectively reviewed 30 patients with locally advanced and nonmetastatic pancreatic cancer who had been treated between 2004 and 2006. Follow-up duration ranged from 4 to 41 months (median, 14.5 months). A total dose of 40 Gy was delivered in 20 fractions using a conventional three-field technique, and then a single fraction of 14, 15, 16, or 17 Gy SBRT was administered as a boost without a break. Twenty-one patients received chemotherapy. Overall and local progression-free survival were calculated and prognostic factors were evaluated. RESULTS: One-year overall survival and local progression-free survival rates were 60.0% and 70.2%, respectively. One patient (3%) developed Grade 4 toxicity. Carbohydrate antigen 19-9 response was found to be an independent prognostic factor for survival. CONCLUSIONS: Our findings indicate that a SBRT boost provides a safe means of increasing radiation dose. Based on the results of this study, we recommend that a well controlled Phase II study be conducted on locally advanced pancreatic cancer.

Stereotactic body radiation therapy in patients with pelvic recurrence from rectal carcinoma.

OBJECTIVE: To investigate the clinical applications of stereotactic body radiation therapy (SBRT) using the CyberKnife system for pelvic recurrence from rectal cancer with a focus on survival and toxicity. METHODS: Between 2002 and 2006, 23 patients with recurrent rectal cancer were treated with SBRT at our institution. The median follow-up was 31 months. Sites of recurrence were pre-sacral in seven patients and the pelvic wall in 16. SBRT doses ranged from 30 to 51 Gy (median 39 Gy) and were delivered in three fractions. Response to treatment was assessed by computed tomography. Overall and local progression-free survival and toxicities were recorded. RESULTS: Four-year overall survival and local control rates were 24.9 and 74.3%, respectively. No prognostic factor was found to affect patient survival or local progression. One patient developed a severe radiation-related toxicity, but recovered completely after treatment. CONCLUSIONS: SBRT for pelvic recurrence was found to be comparable with other modalities with respect to overall survival and complication rates. Further studies are needed to confirm these preliminary results.

Acute arthritis of carpal bones secondary to metastatic gastric cancer.

A 39-year-old woman presented with acute onset of arthralgia in both wrists. Although her primary gastric cancer was in remission after previous treatment, carcinomatous arthritis was suggested by the osteolytic radiographic findings and refractoriness to nonsteroidal anti-inflammatory drugs, which was then confirmed by synovial fluid cytopathology. In view of the high incidence of gastric cancer in Korea, gastric cancer involving the carpal bones should be considered when we encounter a patient presenting with inflammatory peripheral joint arthritis, which might be the first sign of recurrence.

The efficacy of epirubicin, cisplatin, uracil/tegafur, and leucovorin in patients with advanced biliary tract carcinoma.

BACKGROUND: Advanced biliary tract carcinoma is among the most prevalent fatal diseases in Korea. However, to our knowledge, to date no effective therapeutic modality has been shown to prolong the survival of patients in the inoperable stages of this disease. METHODS: This Phase II study was conducted to determine the efficacy and toxicity of a combined regimen of epirubicin, cisplatin, and uracil/tegafur (UFT) modulated by leucovorin in patients with advanced or recurrent biliary tract carcinoma. RESULTS: Eleven of 40 patients (27.5%) had gallbladder carcinoma, and the remaining patients had tumors arising from other sites in the biliary tract. All patients were treated with intravenous epirubicin (50 mg/m(2) on Day 1), intravenous cisplatin (60 mg/m(2) on Day 1), oral UFT (300 mg/m(2) per day on Days 1-21), and oral leucovorin (75 mg per day on Days 1-21). Nine patients exhibited a partial response, representing 22.5% of the possible response rate (95% confidence interval [95% CI], 12.8-32.2%) based on an intention-to-treat analysis. The median survival was 34 weeks (95% CI, 20-48 weeks), and the median time to disease progression was 16 weeks (95% CI, 7-25 weeks). Neutropenia and thrombocytopenia comprised dose-limiting toxicity conditions. CONCLUSIONS: The combination of epirubicin, cisplatin, and UFT modulated by leucovorin was active marginally in patients with advanced biliary tract carcinoma and was capable of stabilizing the disease effectively. Because it was a safe and convenient treatment modality, it may be used in outpatient care with only minor toxicity in patients with advanced malignancies of the biliary tract.