The role of a moisture-barrier latex in controlling retention, stability and release of D-limonene from complex coacervated matrix microparticles formed during spray drying.

Limonene from citrus peel oil is valued as fragrance and flavor additives in food and beverages; however, D-limonene is highly volatile and oxygen-sensitive, thus present storage and stability challenges in food products. A novel, industrially-scalable microencapsulation by in situ complex coacervation during spray drying process (CoCo process) was applied to encapsulate limonene in alginate-gelatin matrix microparticles. Specifically, we investigated the potential to improve upon prior work demonstrating volatile retention and enteric release of limonene from the complex coacervated (CoCo) microcapsules by incorporating ethylcellulose to improve moisture and oxygen barrier properties of the encapsulation matrix. We hypothesized that ethylcellulose, commonly used as a water-barrier coating with pharmaceuticals, would enhance the ability of CoCo microcapsules to retain and shelf-stabilize limonene. The CoCo process alone could achieve limonene retention of 77.7% ± 1.3% during spray drying, with only ∼10% limonene loss and low oxidation rate after 3-weeks of storage in ambient conditions. Contrary to expectations, incorporating ethylcellulose with the CoCo formulation increased volatile losses of limonene during spray drying and during prolonged storage. Moreover, CoCo powders with ethylcellulose accelerated limonene release in water and simulated gastric fluid, and decelerated release in simulated intestinal fluid-a result that was contrary to targeting enteric release. Instead of simply forming a protective water barrier film in the microparticles during spray drying as envisioned, ethylcellulose appeared to bring limonene to the particle surfaces, thereby enhancing volatile losses, facilitating oxidation and accelerating release in acidic aqueous media. Using ethylcellulose as a model, this study demonstrated the potential to formulate CoCo microparticles using latex excipients to control burst release of the payload followed by long-lasting sustained release in air and in aqueous environments.

A New TGF-ß1 Inhibitor, CTI-82, Antagonizes Epithelial-Mesenchymal Transition through Inhibition of Phospho-SMAD2/3 and Phospho-ERK.

Transforming growth factor-ß1 (TGF-ß1) is highly expressed in the tumor microenvironment and known to play a multifunctional role in cancer progression. In addition, TGF-ß1 promotes metastasis by inducing epithelial-mesenchymal transition (EMT) in a variety of tumors. Thus, inhibition of TGF-ß1 is considered an important strategy in the treatment of cancer. In most tumors, TGF-ß1 signal transduction exhibits modified or non-functional characteristics, and TGF-ß1 inhibitors have various inhibitory effects on cancer cells. Currently, many studies are being conducted to develop TGF-ß1 inhibitors from non-toxic natural compounds. We aimed to develop a new TGF-ß1 inhibitor to suppress EMT in cancer cells. As a result, improved chalcone-like chain CTI-82 was identified, and its effect was confirmed in vitro. We showed that CTI-82 blocked TGF-ß1-induced EMT by inhibiting the cell migration and metastasis of A549 lung cancer cells. In addition, CTI-82 reduced the TGF-ß1-induced phosphorylation of SMAD2/3 and inhibited the expression of various EMT markers. Our results suggest that CTI-82 inhibits tumor growth, migration, and metastasis.

Therapeutic Efficacy of ABN401, a Highly Potent and Selective MET Inhibitor, Based on Diagnostic Biomarker Test in MET-Addicted Cancer.

The receptor tyrosine kinase c-MET regulates processes essential for tissue remodeling and mammalian development. The dysregulation of c-MET signaling plays a role in tumorigenesis. The aberrant activation of c-MET, such as that caused by gene amplification or mutations, is associated with many cancers. c-MET is therefore an attractive therapeutic target, and inhibitors are being tested in clinical trials. However, inappropriate patient selection criteria, such as low amplification or expression level cut-off values, have led to the failure of clinical trials. To include patients who respond to MET inhibitors, the selection criteria must include MET oncogenic addiction. Here, the efficacy of ABN401, a MET inhibitor, was investigated using histopathologic and genetic analyses in MET-addicted cancer cell lines and xenograft models. ABN401 was highly selective for 571 kinases, and it inhibited c-MET activity and its downstream signaling pathway. We performed pharmacokinetic profiling of ABN401 and defined the dose and treatment duration of ABN401 required to inhibit c-MET phosphorylation in xenograft models. The results show that the efficacy of ABN401 is associated with MET status and they highlight the importance of determining the cut-off values. The results suggest that clinical trials need to establish the characteristics of each sample and their correlations with the efficacy of MET inhibitors.

Ensemble confidence intervals for binomial proportions.

We propose two measures of performance for a confidence interval for a binomial proportion p: the root mean squared error and the mean absolute deviation. We also devise a confidence interval for p based on the actual coverage function that combines several existing approximate confidence intervals. This "Ensemble" confidence interval has improved statistical properties over the constituent confidence intervals. Software in an R package, which can be used in devising and assessing these confidence intervals, is available on CRAN.

Ketoacidosis with euglycemia in a patient with type 2 diabetes mellitus taking dapagliflozin: A case report.

RATIONALE: Dapagliflozin (a sodium-glucose cotransporter-2 [SGLT2] inhibitor) represents the most recently approved class of oral medications for the treatment of type 2 diabetes. Dapagliflozin lowers plasma glucose concentration by inhibiting the renal reuptake of glucose in the proximal renal tubules. In 2015, the US Food and Drug Administration released a warning concerning a potential increased risk of ketoacidosis in patients taking this medication. PATIENT CONCERNS: We present the case of a 23-year-old woman with type 2 diabetes treated with dapagliflozin (10 mg, once a day) for 2 years who presented to the emergency department with abdominal pain. DIAGNOSES: We diagnosed her with severe ketoacidosis with a normal glucose level (177 mg/dL) due to dapagliflozin, accompanying acute pancreatitis due to hypertriglyceridemia. We concluded that the precipitating factor for euglycemic ketoacidosis was pseudomembranous colitis. INTERVENTIONS: She was treated with intravenous infusions of insulin, isotonic saline, and sodium bicarbonate as diabetic ketoacidosis treatment. OUTCOMES: She was in shock with severe metabolic acidosis. After continuous renal replacement therapy, the uncontrolled metabolic ketoacidosis was treated, and she is currently under follow-up while receiving metformin (500 mg, once a day) and short- and long-acting insulins (8 units 3 times and 20 units once a day). LESSONS: We report an unusual case of SGLT2 inhibitor-induced euglycemic ketoacidosis recovered by continuous renal replacement therapy in a patient with type 2 diabetes and recurrent acute pancreatitis due to hypertriglyceridemia. We diagnosed a rare complication of the SGLT2 inhibitor in a patient with type 2 diabetes in whom uncontrolled metabolic ketoacidosis could be effectively managed via continuous renal replacement therapy.

Development of Human Monoclonal Antibody for Claudin-3 Overexpressing Carcinoma Targeting.

Most malignant tumors originate from epithelial tissues in which tight junctions mediate cell-cell interactions. Tight junction proteins, especially claudin-3 (CLDN3), are overexpressed in various cancers. Claudin-3 is exposed externally during tumorigenesis making it a potential biomarker and therapeutic target. However, the development of antibodies against specific CLDN proteins is difficult, because CLDNs are four-transmembrane domain proteins with high homology among CLDN family members and species. Here, we developed a human IgG1 monoclonal antibody (h4G3) against CLDN3 through scFv phage display using CLDN3-overexpressing stable cells and CLDN3-embedded lipoparticles as antigens. The h4G3 recognized the native conformation of human and mouse CLDN3 without cross-reactivity to other CLDNs. The binding kinetics of h4G3 demonstrated a sub-nanomolar affinity for CLDN3 expressed on the cell surface. The h4G3 showed antibody-dependent cellular cytotoxicity (ADCC) according to CLDN3 expression levels in various cancer cells by the activation of FcγRIIIa (CD16a). The biodistribution of h4G3 was analyzed by intravenous injection of fluorescence-conjugated h4G3 which showed that it localized to the tumor site in xenograft mice bearing CLDN3-expressing tumors. These results indicate that h4G3 recognizes CLDN3 specifically, suggesting its value for cancer diagnosis, antibody-drug conjugates, and potentially as a chimeric antigen receptor (CAR) for CLDN3-expressing pan-carcinoma.

Accuracy of three-dimensional cephalograms generated using a biplanar imaging system.

OBJECTIVE: Biplanar imaging systems allow for simultaneous acquisition of lateral and frontal cephalograms. The purpose of this study was to compare measurements recorded on three-dimensional (3D) cephalograms constructed from two-dimensional conventional radiographs and biplanar radiographs generated using a new biplanar imaging system with those recorded on cone-beam computed tomography (CBCT)-generated cephalograms in order to evaluate the accuracy of the 3D cephalograms generated using the biplanar imaging system. METHODS: Three sets of lateral and frontal radiographs of 15 human dry skulls with prominent facial asymmetry were obtained using conventional radiography, the biplanar imaging system, and CBCT. To minimize errors in the construction of 3D cephalograms, fiducial markers were attached to anatomical landmarks prior to the acquisition of radiographs. Using the 3D Ceph™ program, 3D cephalograms were constructed from the images obtained using the biplanar imaging system (3D cephbiplanar), conventional radiography (3D cephconv), and CBCT (3D cephcbct). A total of 34 measurements were obtained compared among the three image sets using paired t-tests and Bland-Altman plotting. RESULTS: There were no statistically significant differences between the 3D cephbiplanar and 3D cephcbct measurements. In addition, with the exception of one measurement, there were no significant differences between the 3D cephcbct and 3D cephconv measurements. However, the values obtained from 3D cephconv showed larger deviations than those obtained from 3D cephbiplanar. CONCLUSIONS: The results of this study suggest that the new biplanar imaging system enables the construction of accurate 3D cephalograms and could be a useful alternative to conventional radiography.