rGO nanomaterial-mediated cancer targeting and photothermal therapy in a microfluidic co-culture platform.

We developed the microfluidic co-culture platform to study photothermal therapy applications. We conjugated folic acid (FA) to target breast cancer cells using reduced graphene oxide (rGO)-based functional nanomaterials. To characterize the structure of rGO-based nanomaterials, we analyzed the molecular spectrum using UV-visible and Fourier-transform infrared spectroscopy (FT-IR). We demonstrated the effect of rGO-FA-based nanomaterials on photothermal therapy of breast cancer cells in the microfluidic co-culture platform. From the microfluidic co-culture platform with breast cancer cells and human umbilical vein endothelial cells (HUVECs), we observed that the viability of breast cancer cells treated with rGO-FA-based functional nanomaterials was significantly decreased after near-infrared (NIR) laser irradiation. Therefore, this microfluidic co-culture platform could be a potentially powerful tool for studying cancer cell targeting and photothermal therapy.

Generation of uniform-sized multicellular tumor spheroids using hydrogel microwells for advanced drug screening.

Even though in vitro co-culture tumor spheroid model plays an important role in screening drug candidates, its wide applications are currently limited due to the lack of reliable and high throughput methods for generating well-defined and 3D complex co-culture structures. Herein, we report the development of a hydrogel microwell array to generate uniform-sized multicellular tumor spheroids. Our developed multicellular tumor spheroids are structurally well-defined, robust and can be easily transferred into the widely used 2D culture substrates while maintaining our designed multicellular 3D-sphere structures. Moreover, to develop effective anti-cancer therapeutics we integrated our recently developed gold-graphene hybrid nanomaterial (Au@GO)-based photothermal cancer therapy into a series of multicellular tumor spheroid co-culture system. The multicellular tumor spheroids were harvested onto a two-dimensional (2D) substrate, under preservation of their three-dimensional (3D) structure, to evaluate the photothermal therapy effectiveness of graphene oxide (GO)-wrapped gold nanoparticles (Au@GO). From the model of co-culture spheroids of HeLa/Ovarian cancer and HeLa/human umbilical vein endothelial cell (HUVEC), we observed that Au@GO nanoparticles displayed selectivity towards the fast-dividing HeLa cells, which could not be observed to this extent in 2D cultures. Overall, our developed uniform-sized 3D multicellular tumor spheroid could be a powerful tool for anticancer drug screening applications.

Development of a theranostic prodrug for colon cancer therapy by combining ligand-targeted delivery and enzyme-stimulated activation.

The high incidence of colorectal cancer worldwide is currently a major health concern. Although conventional chemotherapy and surgery are effective to some extent, there is always a risk of relapse due to associated side effects, including post-surgical complications and non-discrimination between cancer and normal cells. In this study, we developed a small molecule-based theranostic system, Gal-Dox, which is preferentially taken up by colon cancer cells through receptor-mediated endocytosis. After cancer-specific activation, the active drug Dox (doxorubicin) is released with a fluorescence turn-on response, allowing both drug localization and site of action to be monitored. The therapeutic potency of Gal-Dox was also evaluated, both in vivo and ex vivo, thus illustrating the potential of Gal-Dox as a colorectal cancer theranostic with great specificity.

In vivo imaging of ß-galactosidase stimulated activity in hepatocellular carcinoma using ligand-targeted fluorescent probe.

Development of targeted, selective, and noninvasive fluorescent probes for in vivo visualization of tumor-associated overexpressed enzymes are highly anticipated for cancer diagnosis and therapy. Herein, we developed a noninvasive fluorescent probe (DCDHF-ßgal) for the sensitive detection, and in vivo visualization of ß-galactosidase in hepatocyte HepG2 cells and its xenograft model. As a model system for in vivo targeted imaging, DCDHF-ßgal possessing galactose unit selectively target hepatocyte and monitor the ß-galactosidase activity with deep tissue penetration, and low background interference. DCDHF-ßgal was activated by intracellular ß-galactosidases as the driving force for the release of NIR fluorophore, thereby exhibiting ratiometric optical response. Initial fluorescence emission measured at 615 nm was changed to fluorescence at 665 nm upon activation of DCDHF-ßgal with ß-galactosidase. Ratiometric fluorescence detection of ß-galactosidase was also observed in hepatocellular carcinoma cells and tumor xenograft. The noninvasive in vivo optical imaging facilitated by targeted and enzyme-activated imaging agent would be useful in various biomedical and diagnostic applications.

Liposomal Texaphyrin Theranostics for Metastatic Liver Cancer.

Reported here is a new theranostic agent, 1, which consists of a Gd3+-texaphyrin core conjugated to a doxorubicin prodrug via a disulfide bond. Conjugate 1 was designed to undergo cleavage in the presence of glutathione (GSH), a species typically upregulated in cancer cells. As prepared, conjugate 1 displays no appreciable fluorescence. However, when exposed to excess GSH an increase in the fluorescence intensity at 592 nm is observed that is ascribed to release of free doxorubicin. To improve the solubility and enhance the tumor targeting of 1, it was loaded into folate-receptor-targeted liposomes to produce FL-1 (for folate liposome loaded with 1). As inferred from both fluorescence turn on studies and independent HPLC analyses, FL-1 was found to undergo selective uptake and cleavage to release free Dox in the KB and CT26 cell lines, which express folate receptors on the cell surface, relative to the HepG2 and NIH3T3 cell lines, which show low expression of those receptors. FL-1 was found to produce a greater antiproliferative effect in the case of the KB and CT26 cell lines as compared to that in the HepG2 and NIH3T3 cell lines. FL-1 was also found to provide enhanced magnetic resonance imaging in vivo under conditions of T1 contrast in the early stage of metastatic cancer progression. Finally, time-dependent tumor regrowth studies involving both subcutaneous and metastatic liver cancer mouse models revealed that FL-1 is capable of reducing the tumor burden in vivo.

Acid-triggered release of doxorubicin from a hydrazone-linked Gd(3+)-texaphyrin conjugate.

The hydrazone-based Gd(3+)-texaphyrin doxorubicin conjugate 1, releases active doxorubicin at acidic pH values, allowing its components to be followed by two complementary imaging methods, namely Off-On fluorescence enhancement and MR imaging. It thus acts as a promising theranostic agent.

Hypoxia-directed and activated theranostic agent: Imaging and treatment of solid tumor.

Hypoxia, a distinguished feature of various solid tumors, has been considered as a key marker for tumor progression. Inadequate vasculature and high interstitial pressures result in relatively poor drug delivery to these tumors. Herein, we developed an antitumor theranostic agent, 4, which is activated in hypoxic conditions and can be used for the diagnosis and treatment of solid tumors. Compound 4, bearing biotin, a tumor-targeting unit, and SN38, an anticancer drug, proved to be an effective theranostic agent for solid tumors. SN38 plays a dual role: as an anticancer drug for therapy and as a fluorophore for diagnosis, thus avoids an extra fluorophore and limits cytotoxicity. Compound 4, activated in the hypoxic environment, showed high therapeutic activity in A549 and HeLa cells and spheroids. In vivo imaging of solid tumors confirmed the tumor-specific localization, deep tissue penetration and activation of compound 4, as well as the production of a strong anticancer effect through the inhibition of tumor growth in a xenograft mouse model validating it as a promising strategy for the treatment of solid tumors.

Antioxidant effects of cultured wild ginseng root extracts on the male reproductive function of boars and guinea pigs.

The main objective of this study was to investigate the effects of cultured wild ginseng root extracts (cWGRE) on the sperm of boars and the reproductive system of guinea pigs. Firstly, semen collected from boars (n=10) were incubated in 38°C for 1h with xanthine and xanthine oxidase to generate ROS. The cWGRE was added to the sperm culture system to test its antioxidant effect on the boar sperm. The amount of Reactive Oxygen Species (ROS) was measured by a chemiluminescence assay using luminol. The results indicated that the addition of cWGRE to boar sperm culture inhibited xanthine and xanthine oxidase-induced ROS concentrations. Treatment with cWGRE also had a positive effect on maintaining sperm motility. Effects of cWGRE administration on vitamin C-deficient guinea pigs were further investigated. Hartley guinea pigs (n=25) at 8 weeks of age were randomly divided into five groups. With the exception of the positive control group, each group was fed vitamin C-deficient feed for 21days (d). Respective groups were also orally administered cWGRE, ginseng extract, or mixed ginsenosides for 21 days. In comparison to the control group, oral administration of cWGRE reduced (P<0.05) amount of lipid peroxidation and increased (P<0.05) both glutathione peroxidase concentrations and the trolox equivalent antioxidant capacity. In addition, administration of cWGRE induced increases (P<0.05) in body weight, testosterone concentrations, and spermatid populations. The results of the present study support our hypothesis that cWGRE has positive effects on male reproductive functions via suppression of ROS production.

An activatable prodrug for the treatment of metastatic tumors.

Metastatic cancers have historically been difficult to treat. However, metastatic tumors have been found to have high levels of reactive oxygen species such as hydrogen peroxide (H2O2), supporting the hypothesis that a prodrug could be activated by intracellular H2O2 and lead to a potential antimetastatic therapy. In this study, prodrug 7 was designed to be activated by H2O2-mediated boronate oxidation, resulting in activation of the fluorophore for detection and release of the therapeutic agent, SN-38. Drug release from prodrug 7 was investigated by monitoring fluorescence after addition of H2O2 to the cancer cells. Prodrug 7 activated by H2O2, selectively inhibited tumor cell growth. Furthermore, intratracheally administered prodrug 7 showed effective antitumor activity in a mouse model of metastatic lung disease. Thus, this H2O2-responsive prodrug has therapeutic potential as a novel treatment for metastatic cancer via cellular imaging with fluorescence as well as selective release of the anticancer drug, SN-38.

Palatal implants for persistent snoring and mild obstructive sleep apnea after laser-assisted uvulopalatoplasty.

Laser-assisted uvulopalatoplasty (LAUP) was widely performed in 1990s as a surgical therapeutic procedure to improve snoring or mild obstructive sleep apnea (OSA). However, LAUP is not currently recommended as a treatment for OSA because the evidence for its efficacy is insufficient. Little is known about alternative minimally invasive surgery in patients who refuse continuous positive airway pressure or oral appliance after failed LAUP. We present a case of successful surgical treatment of persistent snoring and mild OSA with palatal implants after LAUP. This case suggests that palatal implants may be offered as an alternative surgical procedure for selective patients with persistent or recurrent snoring or mild OSA after LAUP.

An activatable theranostic for targeted cancer therapy and imaging.

A new theranostic strategy is described. It is based on the use of an "all in one" prodrug, namely the biotinylated piperazine-rhodol conjugate 4 a. This conjugate, which incorporates the anticancer drug SN-38, undergoes self-immolative cleavage when exposed to biological thiols. This leads to the tumor-targeted release of the active SN-38 payload along with fluorophore 1 a. This release is made selective as the result of the biotin functionality. Fluorophore 1 a is 32-fold more fluorescent than prodrug 4 a. It permits the delivery and release of the SN-38 payload to be monitored easily in vitro and in vivo, as inferred from cell studies and ex vivo analyses of mice xenografts derived from HeLa cells, respectively. Prodrug 4 a also displays anticancer activity in the HeLa cell murine xenograft tumor model. On the basis of these findings we suggest that the present strategy, which combines within a single agent the key functions of targeting, release, imaging, and treatment, may have a role to play in cancer diagnosis and therapy.

Comparison of diagnostic accuracy between CellprepPlus® and ThinPrep® liquid-based preparations in effusion cytology.

Liquid-based cytology (LBC) is being increasingly used for body fluid specimens and has improved diagnostic accuracy when compared to conventional smears. We compared the diagnostic accuracy and cellular morphologic features between CellprepPlus® LBC and ThinPrep® LBC in effusion cytology. One hundred and eighty body fluid specimens, consisting of 119 pleural fluid specimens, 59 peritoneal fluid specimens, and 2 pericardial fluid specimens, were obtained from 166 patients. Equal volumes of body fluid from each specimen were used in the CellprepPlus® and ThinPrep® preparations. Sensitivity, specificity, and positive and negative predictive values were evaluated. In addition, we selected 16 specimens from patients with metastatic adenocarcinoma, confirmed them by both LBC preparations, and measured the size of the nucleus in the tumor cells in these specimens. The sensitivity of the CellprepPlus® and ThinPrep® methods was 73.1% and 50.0%, respectively. The specificity and positive predictive values were 100% for both LBC methods, and the negative predictive values of the CellprepPlus® and ThinPrep® methods were 90.9% and 83.3%, respectively. The average nuclear size of the tumor cells was calculated as 20.87 µm using the CellprepPlus® method and 15.08 µm using the ThinPrep® method (P < 0.05). The CellprepPlus® method provided better diagnostic accuracy of effusion cytology compared to the ThinPrep® method and revealed the characteristic morphological features of tumor cells, including large and hypochromatic nuclei, prominent nucleoli, distinct nuclear membranes, and high cellularity.

Association of protein expression in isolated milk epithelial cells and cis-9, trans-11 conjugated linoleic acid proportions in milk from dairy cows.

BACKGROUND: The effects of the individual variation among dairy cows on the synthesis of cis-9, trans-11 conjugated linoleic acid (CLA) are still not well characterised. Therefore, the protein expression profiles of isolated milk epithelial cells (MECs) were detected by two-dimensional electrophoresis and their correlation with the various proportion of cis-9, trans-11 CLA were evaluated. RESULTS: Although animals were offered the same diet, the proportion of cis-9, trans-11 CLA in group High (1.02 ± 0.10%) was twice as high as that in group Low (0.59 ± 0.14%) (P < 0.05). MECs with the characteristics of native epithelial cells were successfully isolated from the milk and these cells had no obvious RNA degradation or were hardly contaminated with leucocytes or blood red cells. Moreover, the protein expression pattern of cathelicidin 5 in isolated MECs was positive, whereas annexin I (confirmed by real-time polymerase chain reaction), ZW10 interactor and κ-casein were negatively related to the proportion of cis-9, trans-11 CLA in the milk fat. CONCLUSION: The varied individual content of cis-9, trans-11 CLA in cows may be associated with annexin I. These findings may provide some theoretical basis for studies concerning the effects of the individual variation among dairy cows of the synthesis of cis-9, trans-11 CLA. © 2013 Society of Chemical Industry.

Tumor bed volumetric changes during breast irradiation for the patients with breast cancer.

PURPOSE: The aim of this study was to evaluate changes in breast tumor bed volume during whole breast irradiation (WBI). MATERIALS AND METHODS: From September 2011 to November 2012, thirty patients who underwent breast-conserving surgery (BCS) followed by WBI using computed tomography (CT) simulation were enrolled. Simulation CT scans were performed before WBI (CT1) and five weeks after the breast irradiation (CT2). The tumor bed was contoured based on surgical clips, seroma, and postoperative change. We retrospectively analyzed the factors associated with tumor bed volumetric change. RESULTS: The median tumor bed volume on CT1 and CT2 was 29.72 and 28.6 mL, respectively. The tumor bed volume increased in 9 of 30 patients (30%) and decreased in 21 of 30 patients (70%). The median percent change in tumor bed volume between initial and boost CT was -5%. Seroma status (p = 0.010) was a significant factor in tumor bed volume reduction of 5% or greater. However, patient age, body mass index, palpability, T stage, axillary lymph node dissection, and tumor location were not significant factors for tumor bed volumetric change. CONCLUSION: In this study, volumetric change of tumor bed cavity was frequent. Patients with seroma after BCS had a significant volume reduction of 5% or greater in tumor bed during breast irradiation. Thus, resimulation using CT is indicated for exquisite boost treatment in breast cancer patients with seroma after surgery.

Evaluation of Urine Cytology in Urothelial Carcinoma Patients: A Comparison of CellprepPlus® Liquid-Based Cytology and Conventional Smear.

BACKGROUND: Urine cytology is an important test in the screening of urothlelial neoplasms. The conventional smear (CS) method of testing urine samples has a low sensitivity, approximately 50% result accuracy for detecting urothelial carcinomas, while liquid-based cytology (LBC) has much improved diagnostic accuracy, sensitivity, and specificity. The aim of this study was to compare the morphologic features and diagnostic efficacy of CellprepPlus® LBC with those of CS for urine cytology. METHODS: A total of 713 cases of urine specimens collected from November 2009 to September 2010 were included. All specimens were divided equally for the preparation of CellprepPlus® LBC and CS for each case. RESULTS: CellprepPlus® revealed more cellularity, a cleaner background and better cytomorphologic features, but it showed a less intact architectural pattern compared to that of CS. Of the 88 histologically confirmed cases, the diagnostic sensitivity for CellprepPlus® was 50% and higher than the 37.5% for CS. The specificity of both preparations was 100%. CONCLUSIONS: The CellprepPlus® showed an improved quality of slides and provided better diagnostic accuracy, thus CellprepPlus® could be a first-line screening tool in urinary tract cytology.

A Prospective Population-based Study of Total Nasal Resistance in Korean Subjects.

OBJECTIVES: Rhinomanometry is a widely accepted method for objective assessment of nasal patency. However, few studies have reported the values of otherwise healthy population for nasal resistance in East Asians. The purpose of this study was to measure normal total nasal resistance (TNR) values in a large sample of Korean adults and to reveal parameters contributing to TNR values. METHODS: Subjects were enrolled from a cohort of the Korean Genome and Epidemiology Study. They were evaluated by anthropometry, questionnaire, and active anterior rhinomanometry at transnasal pressures of 100 and 150 Pascal (Pa). RESULTS: The study sample consisted of 2,538 healthy subjects (1,298 women and 1,240 men) aged 20 to 80 years. Normal reference TNR values were 0.19±0.08 Pa/cm(3)/second at 100 Pa and 0.22±0.09 Pa/cm(3)/second at 150 Pa. The TNR of women was significantly higher than that of men (P<0.0001). TNR decreased with increasing age in both genders (P<0.05). In women, lower body weight was related to increasing TNR. In men, current smokers had higher TNR than ex-smokers and never smokers. CONCLUSION: The results of the present study provide information regarding the values of otherwise healthy population of TNR and parameters associated with TNR in Korean adults.

Magnetic resonance imaging of superparamagnetic iron oxide-labeled macrophage infiltrates in acute-phase renal ischemia-reperfusion mouse model.

Macrophages play a key role in the initial pathogenesis of kidney ischemia-reperfusion (I-R) injury, but the mechanism of their spatial and temporal recruitment from circulation remains uncertain. This study aimed to evaluate the feasibility of magnetic resonance imaging (MRI) for detecting intravenously administered superparamagnetic iron oxide (SPIO)-labeled macrophages in an experimental renal I-R mouse model. Unilateral kidney I-R mice were imaged with a 4.7-T MRI scanner before and after administration of SPIO-labeled macrophages (RAW 264.7). On MR images, adoptive transfer of SPIO-labeled macrophages in the acute phase (1-2 days after I-R) caused a band-shaped signal-loss zone resulting from macrophage infiltrations, in the outer medullary region of injured kidneys. MRI detection of macrophages homing to an injured kidney may facilitate early detection and investigation of the pathogenesis of acute kidney injury and be a strategy for determining the treatment of acute renal failure. From the Clinical Editor: This study evaluated the feasibility of magnetic resonance imaging for detecting superparamagnetic iron oxide (SPIO)-labeled macrophages in a renal ischemia-reperfusion mouse model. Similar strategies in humans may facilitate early detection and stratification of acute kidney injury.

Effectiveness of nasal surgery alone on sleep quality, architecture, position, and sleep-disordered breathing in obstructive sleep apnea syndrome with nasal obstruction.

BACKGROUND: The goal of this study was to evaluate the effect of nasal surgery alone on sleep quality, architecture, position, and sleep-disordered breathing (SDB) (including obstructive apnea and snoring) in adult patients with obstructive sleep apnea syndrome (OSAS) and nasal obstruction. METHODS: A total of 22 consecutive male patients (mean age, 41.3 ± 10.9 years) with OSAS and nasal obstruction, who underwent nasal surgery alone, were enrolled in the study. We compared polysomnographic data related with sleep quality (sleep efficiency [SE] and arousal index [ArI]), sleep architecture (stages N [nonrapid eye movement], 1, 2, and 3, and R [rapid eye movement]), the distribution of sleep positions, and SDB (apnea-hypopnea index [AHI], apnea index [AI], minimum arterial oxygen saturation [SaO(2)], and snoring) before and after nasal surgery. RESULTS: SE (from 86.6 ± 6.3% to 89.7 ± 7.1%; p = 0.039), stage R (from 15.3 ± 4.8% to 18.6 ± 5.4%; p = 0.016) and snoring (from 32.2 ± 16.4% to 25.8 ± 18.6%; p = 0.045) significantly changed after nasal surgery. However, there were no significant changes in ArI, other sleep stages, the proportion of sleep time spent in the supine position, AHI, AI, and minimum SaO(2) after nasal surgery. CONCLUSION: Nasal surgery alone was partially effective in improving sleep quality, architecture, and snoring, but it had no effect on the change of the distribution of sleep positions and obstructive apnea in patients with OSAS and nasal obstruction.

Efficacy of single-staged modified uvulopalatopharyngoplasty with nasal surgery in adults with obstructive sleep apnea syndrome.

OBJECTIVE: The aim of this study was to investigate the clinical efficacy of single-staged modified uvulopalatopharyngoplasty (UPPP) with nasal surgery and the relationship between its surgical outcomes and an anatomy-based staging system in patients with obstructive sleep apnea syndrome (OSAS) with nasal obstruction. STUDY DESIGN AND SETTING: Before-after analysis at a university hospital. SUBJECTS AND METHODS: A total of 41 consecutive OSAS patients (mean age 40.1 ± 7.3 years) who underwent single-staged modified (uvula-preserving) UPPP with nasal surgery were included. The investigators compared subjective symptoms and polysomnographic data before and after surgery and investigated objective surgical outcomes according to the anatomy-based (Friedman) staging system and postoperative complications. Surgical success was defined as a reduction of at least 50% in preoperative apnea-hypopnea index (AHI) and a postoperative AHI of less than 20 per hour. RESULTS: After simultaneous nasal-oropharyngeal surgery, the AHI significantly decreased (from 45.9 ± 23.4 to 20.9 ± 22.1 events per hour; P < .001) and the overall success rate was 56.1% (23/41). Surgical success rates in stages I, II, and III were 70.6% (12/17), 60.0% (9/15), and 22.2% (2/9), respectively. There were no major complications during or after surgery, and most minor complications were transient and resolved without morbidity. CONCLUSIONS: Single-staged modified UPPP with nasal surgery is an available and relatively safe surgical approach in OSAS patients with nasal obstruction. To achieve the best possible surgical outcomes, it is important to select appropriate patients using the anatomy-based staging system.

Effect of successful surgical treatment on changes of position during sleep in adults with obstructive sleep apnea syndrome.

OBJECTIVES: The purpose of this study was to evaluate the changes of position during sleep as determined by polysomnography before and after upper airway surgery for obstructive sleep apnea syndrome in patients with no response to surgery ("nonresponse group") and in those who did have a response to surgery ("response group"). METHODS: We analyzed a total of 106 polysomnograms from 53 subjects and compared the preoperative-postoperative differences in the frequency of positional changes during sleep and the distribution of sleep positions between the nonresponse group (n = 25) and the response group (n = 28). Surgical response was defined as a greater than 50% decrease in the postoperative apnea-hypopnea index. RESULTS: The positional change index in the response group was significantly reduced (from 4.2 +/- 3.8 to 2.6 +/- 1.6; p = 0.038), whereas the positional change index in the nonresponse group did not significantly change (from 3.4 +/- 2.0 to 3.4 +/- 2.1; p = 0.861). The proportion of sleep time spent in the supine position did not significantly change in the nonresponse group (from 62.4% +/- 18.1% to 60.5% +/- 21.3%; p = 0.904) or the response group (from 55.5% +/- 23.9% to 60.1% +/- 23.1%; p = 0.412). CONCLUSIONS: The frequency of positional changes during sleep was significantly decreased with the improvement of respiratory disturbances and arousals in the response group after upper airway surgery.