High-throughput organo-on-pillar (high-TOP) array system for three-dimensional ex vivo drug testing.

The development of organoid culture technologies has triggered industrial interest in ex vivo drug test-guided clinical response prediction for precision cancer therapy. The three-dimensional culture encapsulated with basement membrane (BM) components is extremely important in establishing ex vivo organoids and drug sensitivity tests because the BM components confer essential structures resembling tumor histopathology. Although numerous studies have demonstrated three-dimensional culture-based drug screening methods, establishing a large-scale drug-screening platform with matrix-encapsulated tumor cells is challenging because the arrangement of microspots of a matrix-cell droplet onto each well of a microwell plate is inconsistent and difficult to standardize. In addition, relatively low scales and lack of reproducibility discourage the application of three-dimensional organoid-based drug screening data for precision treatment or drug discovery. To overcome these limitations, we manufactured an automated organospotter-integrated high-throughput organo-on-pillar (high-TOP) drug-screening platform. Our system is compatible with various extracellular matrices, including BM extract, Matrigel, collagen, and hydrogel. In addition, it can be readily utilized for high-content analyses by simply exchanging the bottom plates without disrupting the domes. Our system demonstrated considerable robustness, consistency, reproducibility, and biological relevancy in three-dimensional drug sensitivity analyses using Matrigel-encapsulated ovarian cancer cell lines. We also demonstrated proof-of-concept cases representing the clinical feasibility of high-TOP-assisted ex vivo drug tests linked to clinical chemo-response in ovarian cancer patients. In conclusion, our platform provides an automated and standardized method for ex vivo drug-sensitivity-guided clinical response prediction, suggesting effective chemotherapy regimens for patients with cancer.

Ecklonia cava Extract and Dieckol Attenuate Cellular Lipid Peroxidation in Keratinocytes Exposed to PM10.

Airborne particulate matter can cause oxidative stress, inflammation, and premature skin aging. Marine plants such as Ecklonia cava Kjellman contain high amounts of polyphenolic antioxidants. The purpose of this study was to examine the antioxidative effects of E. cava extract in cultured keratinocytes exposed to airborne particulate matter with a diameter of <10 µm (PM10). After the exposure of cultured HaCaT keratinocytes to PM10 in the absence and presence of E. cava extract and its constituents, cell viability and cellular lipid peroxidation were assessed. The effects of eckol and dieckol on cellular lipid peroxidation and cytokine expression were examined in human epidermal keratinocytes exposed to PM10. The total phenolic content of E. cava extract was the highest among the 50 marine plant extracts examined. The exposure of HaCaT cells to PM10 decreased cell viability and increased lipid peroxidation. The PM10-induced cellular lipid peroxidation was attenuated by E. cava extract and its ethyl acetate fraction. Dieckol more effectively attenuated cellular lipid peroxidation than eckol in both HaCaT cells and human epidermal keratinocytes. Dieckol and eckol attenuated the expression of inflammatory cytokines such as tumor necrosis factor- (TNF-) α, interleukin- (IL-) 1ß, IL-6, and IL-8 in human epidermal keratinocytes stimulated with PM10. This study suggested that the polyphenolic constituents of E. cava, such as dieckol, attenuated the oxidative and inflammatory reactions in skin cells exposed to airborne particulate matter.

Effect of selenium supplementation for protection of salivary glands from iodine-131 radiation damage in patients with differentiated thyroid cancer.

OBJECTIVE: In the current study, we examined whether selenium supplementation during iodine-131 (131I) treatment had a radio-protective effect on salivary glands. SUBJECTS AND METHODS: Sixteen patients with differentiated thyroid cancer were prospectively enrolled in the study. Patients after total thyroidectomy, before 131I treatment, were divided into two groups; 8 patients in the selenium group and 8 patients in the control group. Patients in the selenium group received 300νg of selenium orally for 10 days, from 3 days before to 6 days after 131I treatment. The control group received a placebo over the same period. To assess salivary gland function, salivary gland scintigraphy was performed before and 6 months after 131I treatment. Serum amylase and whole blood selenium levels were measured before and 2 days and 6 months after 131I treatment. Using salivary gland scintigraphy, maximum uptake ratio (MUR), maximum secretion percentage (MSP), and ejection fraction (EF) of each salivary gland were calculated. RESULTS: Baseline clinical characteristics, baseline amylase and selenium levels, and parameters of baseline salivary gland scintigraphy were not significantly different between selenium and control groups (P>0.05). On a blood test performed 2 days after 131I treatment, the selenium group showed a significantly higher whole blood selenium level (P=0.008) and significantly lower serum amylase level (P=0.009) than the control group. On follow-up salivary gland scintigraphy, the control group showed significantly decreased, MUR of the bilateral parotid and left submandibular glands, MSP of the bilateral parotid and submandibular glands, and EF of the left submandibular glands (P<0.05), while the selenium group only had a significant decrease in MSP of the right submandibular gland and EF of the left submandibular gland (P<0.05). CONCLUSION: Selenium supplementation during 131I treatment was effective to reduce salivary glands damage by 131I radiation in patients with differentiated thyroid cancer.

Patient-Derived Xenograft Models of Epithelial Ovarian Cancer for Preclinical Studies.

PURPOSE: Patient-derived tumor xenografts (PDXs) can provide more reliable information about tumor biology than cell line models. We developed PDXs for epithelial ovarian cancer (EOC) that have histopathologic and genetic similarities to the primary patient tissues and evaluated their potential for use as a platform for translational EOC research. MATERIALS AND METHODS: We successfully established PDXs by subrenal capsule implantation of primary EOC tissues into female BALB/C-nude mice. The rate of successful PDX engraftment was 48.8% (22/45 cases). Hematoxylin and eosin staining and short tandem repeat analysis showed histopathological and genetic similarity between the PDX and primary patient tissues. RESULTS: Patients whose tumors were successfully engrafted in mice had significantly inferior overall survival when compared with those whose tumors failed to engraft (p=0.040). In preclinical tests of this model, we found that paclitaxel-carboplatin combination chemotherapy significantly deceased tumor weight in PDXs compared with the control treatment (p=0.013). Moreover, erlotinib treatment significantly decreased tumor weight in epidermal growth factor receptor-overexpressing PDX with clear cell histology (p=0.023). CONCLUSION: PDXs for EOC with histopathological and genetic stability can be efficiently developed by subrenal capsule implantation and have the potential to provide a promising platform for future translational research and precision medicine for EOC.

Analysis of Geometric Shifts and Proper Setup-Margin in Prostate Cancer Patients Treated With Pelvic Intensity-Modulated Radiotherapy Using Endorectal Ballooning and Daily Enema for Prostate Immobilization.

We evaluate geometric shifts of daily setup for evaluating the appropriateness of treatment and determining proper margins for the planning target volume (PTV) in prostate cancer patients.We analyzed 1200 sets of pretreatment megavoltage-CT scans that were acquired from 40 patients with intermediate to high-risk prostate cancer. They received whole pelvic intensity-modulated radiotherapy (IMRT). They underwent daily endorectal ballooning and enema to limit intrapelvic organ movement. The mean and standard deviation (SD) of daily translational shifts in right-to-left (X), anterior-to-posterior (Y), and superior-to-inferior (Z) were evaluated for systemic and random error.The mean ±â€ŠSD of systemic error (Σ) in X, Y, Z, and roll was 2.21 ±â€Š3.42 mm, -0.67 ±â€Š2.27 mm, 1.05 ±â€Š2.87 mm, and -0.43 ±â€Š0.89°, respectively. The mean ±â€ŠSD of random error (δ) was 1.95 ±â€Š1.60 mm in X, 1.02 ±â€Š0.50 mm in Y, 1.01 ±â€Š0.48 mm in Z, and 0.37 ±â€Š0.15° in roll. The calculated proper PTV margins that cover >95% of the target on average were 8.20 (X), 5.25 (Y), and 6.45 (Z) mm. Mean systemic geometrical shifts of IMRT were not statistically different in all transitional and three-dimensional shifts from early to late weeks. There was no grade 3 or higher gastrointestinal or genitourianry toxicity.The whole pelvic IMRT technique is a feasible and effective modality that limits intrapelvic organ motion and reduces setup uncertainties. Proper margins for the PTV can be determined by using geometric shifts data.

Clinical significance of diffuse hepatic uptake on post-therapeutic early and delayed (131)I scan in differentiated thyroid cancer: a preliminary report.

OBJECTIVE: The purpose of this study was to investigate the clinical significance of diffuse hepatic uptake on post-therapeutic early and delayed (131)I scan in patients with differentiated thyroid cancer (DTC). METHODS: We retrospectively analyzed 219 DTC patients who underwent high-dose (131)I treatment and subsequent post-therapeutic dual (131)I scan. Both early (third day after (131)I treatment) and delayed (5-6th day after (131)I treatment) (131)I scan images were visually assessed and diffuse hepatic uptake was scored using a 4-point grading system depending on intensity. RESULTS: On early (131)I scan, 73 patients (33.4 %) showed diffuse hepatic uptake, while 191 patients (87.2 %) patients showed diffuse hepatic uptake on delayed scan (p < 0.0001). The serum levels of ALT in patients with diffuse hepatic uptake on early scan were higher than those without diffuse hepatic uptake on early scan (p = 0.03 for ALT and p = 0.08 for AST). The serum levels of ALT and AST trended with the grade of hepatic uptake on delayed scan (p = 0.03 for ALT and p = 0.05 for AST). Diffuse hepatic uptake on early or delayed scan showed no significant relationship in the presence of thyroid remnants, metastatic DTC lesions, tumor recurrence during follow-up, and the serum thyroglobulin level (p > 0.05). On logistic regression analysis, both serum ALT (p = 0.01) and AST (p = 0.04) levels were significant predictive factors for diffuse hepatic uptake on early scan, while only serum ALT (p = 0.01) level was significant predictive factor for diffuse hepatic uptake on delayed scan. CONCLUSIONS: The frequency of diffuse hepatic uptake on the delayed scan was significantly higher than the early scan. Diffuse hepatic uptake on early post-therapeutic scan and the intensity of diffuse hepatic uptake on delayed scan showed significant correlation with the serum levels of hepatic enzymes, but no significant association in the presence of thyroid remnants, metastatic DTC lesions, and tumor recurrence during follow-up. The timing and intensity of diffuse hepatic uptake on post-therapeutic scan may be related with factors such as hepatic function other than the thyroid tissue or DTC.

Platinum-based combination chemotherapy vs. weekly cisplatin during adjuvant CCRT in early cervical cancer with pelvic LN metastasis.

AIM: Adjuvant concurrent chemoradiation (CCRT) should be considered in surgically-treated patients with early-stage cervical cancer (ECC) who exhibit pelvic lymph node (LN) metastasis. Platinum-based chemotherapy is usually recommended during adjuvant CCRT, however, it is unclear which regimen has better prognostic outcomes. PATIENTS AND METHODS: We reviewed the electronic medical records to find patients with primary ECC (FIGO stages IB-IIA) who underwent type III radical hysterectomy and adjuvant CCRT due to pelvic LN metastasis at the Samsung Medical Center, Sungkyunkwan University School of Medicine in Seoul, Korea, from November 1997 to September 2007. RESULTS: Among 75 patients, 34 received weekly cisplatin. Combination chemotherapy was performed without consolidation in 21 patients and with consolidation in 20 patients. The mean follow-up period was 59.0 months and the five-year survival rate was 84.4%. In multivariate analysis, combination chemotherapy with and without consolidation was associated with improved disease-free survival [hazard ratio (HR)=0.23, 95% confidence interval (CI)=0.06-0.88, p=0.032, and HR=0.29, 95% CI=0.09-0.91, p=0.034, respectively]; combination chemotherapy with consolidation significantly improved overall survival (HR=0.11, 95% CI=0.02-0.87, p=0.037) when compared to weekly cisplatin. CONCLUSION: We found that platinum-based combination chemotherapy during adjuvant CCRT after surgery promoted better survival than a weekly cisplatin regimen in ECC patients with pelvic LN metastasis.

Clinical utility of 18F-FDG PET/CT concurrent with 131I therapy in intermediate-to-high-risk patients with differentiated thyroid cancer: dual-center experience with 286 patients.

UNLABELLED: Patients with differentiated thyroid carcinoma (DTC) are treated with (131)I therapy after total thyroidectomy or surgical resection of recurrent tumor. However, some recurrent DTC lesions are not iodine-avid, which affects further treatment planning. The aim of this study was to evaluate the clinical benefit of (18)F-FDG PET/CT performed concurrently with (131)I therapy in DTC patients with intermediate to high risk. METHODS: We retrospectively enrolled 286 DTC patients at 2 Korean medical centers who comprised 2 different patient groups: 28 patients who underwent adjuvant (131)I treatment after curative surgical resection of recurrent tumor and 258 patients with intermediate to high risk who underwent (131)I ablation after total thyroidectomy. (131)I therapy and (18)F-FDG PET/CT scanning were performed on the same day. Administration of l-thyroxine was withheld from all enrollees for 4 wk before (131)I treatment. RESULTS: In 39 patients (14%), (18)F-FDG PET/CT detected additional recurrent or metastatic lesions that were not detected on the posttherapy (131)I scan, and the treatment plan was changed for 30 patients (10%) based on such findings. Among the 28 patients receiving (131)I treatment after resection of recurrent tumor, PET/CT detected additional lesions in 46%, and treatment was changed in 43%. Assessing a subgroup of stage T3-T4N1 patients with tumor size > 2.0 cm, among 258 patients undergoing (131)I ablation after total thyroidectomy, we found that 25% had additional positive PET/CT results, and treatment changed for 17%. In contrast, 8% of stage T3-T4N1 patients with tumor size ≤ 2.0 cm, 6% of stage T1-T2N1 patients, and 3% of stage T3-T4N0 patients had additional positive PET/CT findings. CONCLUSION: (18)F-FDG PET/CT performed concurrently with (131)I therapy detected additional lesions in 14% of DTC patients and was particularly helpful for detecting additional lesions in patients undergoing (131)I therapy after resection of recurrent tumor or in stage T3-T4N1 patients with tumor size > 2.0 cm.

Modified yukmijihwangtang suppresses the production of proinflammatory cytokines in the intravesical hydrochloric acid-induced cystitis rat model via the NF-κB pathway.

Yukmijihwangtang (YM), a boiled extract of medicinal plants, has been prescribed for patients with kidney dysfunction in Korea; however, the mechanism underlying its therapeutic effects has not been fully elucidated. This study was conducted to evaluate the beneficial effects on bladder function by using modified YM (M-YM), which included Ulmi radicis cortex in addition to the six traditional medicinal plants in YM. Bladder irritation of the rats was caused by intravesical instillation of HCl. The animals were divided into six groups: sham group, cystitis-injury group with no treatment, cystitis-injury group with prednisolone treatment (5 mg/kg), and cystitis-injury with M-YM treatment (100, 200 or 500 mg/kg groups). Whole bladders were collected at day eight after injury. Samples were analyzed by histological and immunological examinations. An in vitro study was performed to determine whether M-YM extracts inhibit lipopolysaccharide (LPS)-induced nitric oxide (NO) production and IκB phosphorylation in a human uroepithelial cell line of T24 cells. Administration of M-YM notably improved bladder histological changes, and suppressed IL-6/TNF α production and IκB phosphorylation in a rat model of chronic cystitis. M-YM also inhibited LPS-induced NO production and IκB phosphorylation in T24 cells. This study suggests that administration of M-YM might be an applicable therapeutic traditional medicine for the treatment of interstitial cystitis.

The prognostic significance of tumor volume regression during radiotherapy and concurrent chemoradiotherapy for cervical cancer using MRI.

OBJECTIVE: The purpose of this study was to determine the prognostic significance of tumor volume regression rate during radiotherapy (RT) measured by three serial magnetic resonance imaging (MRIs) studies performed in patients treated with RT alone and compare the results with patients treated with concurrent chemoradiotherapy (CCRT). METHODS: We evaluated 81 patients with uterine cervical cancer who underwent three serial MR examinations, i.e., at the start of RT, at 36-45 Gy of external RT and 1 month after the end of RT. Forty-three patients were treated with RT alone and 38 patients were treated with CCRT. Pre-RT tumor volume (V1), the tumor volume regression rate measured during the fourth week of RT and residual tumor volume at 1 month after the end of RT (V3) were determined for each patient. The cut-off value used for the three parameters studied was the one that made the largest outcome difference. These volume parameters were analyzed to determine a difference in the treatment outcome. RESULTS: In the patients treated with CCRT, the mean value of the V1 was larger and the mean value of the V3 was smaller than in patients treated with RT alone. The mean value of the mid-RT regression rate was somewhat higher in patients treated with CCRT than in patients treated with RT alone; however, this difference was not statistically significant (79% vs. 69%). In both the RT alone and the CCRT group, the patients with a mid-RT regression >/=75% had 100% 5-year local control rates and a better disease free survival than the patients with mid-RT regression <75%. The patients with V3=0 cm(3) also had a better 5-year local control rate than the patients with a V3>0 cm(3), but statistical significance was found only in the patients treated with CCRT. CONCLUSIONS: The mid-RT tumor volume regression rate, at 36-45 Gy of external RT, was a predictor of local control rate in both RT and CCRT patient groups. However, in the patients who were treated with CCRT, the local control rate difference was even larger by post-RT residual volume than by the mid-RT tumor regression rate. Further studies on appropriate evaluation timing for mid-RT response in patients receiving CCRT are needed.

Phase II study of consolidation chemotherapy after concurrent chemoradiation in cervical cancer: preliminary results.

PURPOSE: Our aim was to determine the efficacy of consolidation chemotherapy after concurrent chemoradiation (CCRT) using high-dose-rate brachytherapy in patients with locally advanced cervical carcinoma. METHODS AND MATERIALS: Patients with cervical carcinoma (FIGO stage IB2-IVA) were treated with external beam radiation therapy to the whole pelvis (50.4 Gy) and high-dose-rate brachytherapy (24 Gy to point A). Cisplatin 60 mg/m(2) (Day 1) and 5-fluorouracil 1000 mg/m(2) (Days 1-5) were given every 3 weeks starting concurrently with the radiation and followed by 3 more cycles of consolidation for a total of 6 cycles. RESULTS: Thirty patients (94%) received 3 more cycles of post-CCRT consolidation chemotherapy and were evaluable for the toxicity and efficacy of consolidation. The most common toxicities of Grade 2 or higher were nausea or vomiting (47%) and anemia (33%). Late complications of the rectum and bladder occurred in 13% and 6% of the patients, respectively. The clinical complete response rate was 87% (95% CI, 75%-99%). During a median follow-up of 27 months (range, 6-58 months), 5 patients (17%) had recurrence; the sites of failure were 3 (10%) inside the radiation field and 2 (7%) outside the radiation field. The estimated 3-year progression-free survival rate was 83% (95% CI, 67%-99%) and overall survival rate was 91% (95% CI, 79%-100%). CONCLUSIONS: Consolidation chemotherapy after CCRT is well tolerated and effective in patients with locally advanced cervical carcinoma. A prospective randomized trial to compare this treatment strategy with standard CCRT seems to be worthwhile.