Quantification of Unencapsulated Drug in Target Tissues Demonstrates Pharmacological Properties and Therapeutic Effects of Liposomal Topotecan (FF-10850).

PURPOSE: Quantifying unencapsulated drug concentrations in tissues is crucial for understanding the mechanisms underlying the efficacy and safety of liposomal drugs; however, the methodology for this has not been fully established. Herein, we aimed to investigate the enhanced therapeutic potential of a pegylated liposomal formulation of topotecan (FF-10850) by analyzing the concentrations of the unencapsulated drug in target tissues, to guide the improvement of its dosing regimen. METHODS: We developed a method for measuring unencapsulated topotecan concentrations in tumor and bone marrow interstitial fluid (BM-ISF) and applied this method to pharmacokinetic assessments. The ratios of the area under the concentration-time curves (AUCs) between tumor and BM-ISF were calculated for total and unencapsulated topotecan. DNA damage and antitumor effects of FF-10850 or non-liposomal topotecan (TPT) were evaluated in an ES-2 mice xenograft model. RESULTS: FF-10850 exhibited a much larger AUC ratio between tumor and BM-ISF for unencapsulated topotecan (2.96), but not for total topotecan (0.752), than TPT (0.833). FF-10850 promoted milder DNA damage in the bone marrow than TPT; however, FF-10850 and TPT elicited comparable DNA damage in the tumor. These findings highlight the greater tumor exposure to unencapsulated topotecan and lower bone marrow exposure to FF-10850 than TPT. The dosing regimen was successfully improved based on the kinetics of unencapsulated topotecan and DNA damage. CONCLUSIONS: Tissue pharmacokinetics of unencapsulated topotecan elucidated the favorable pharmacological properties of FF-10850. Evaluation of tissue exposure to an unencapsulated drug with appropriate pharmacodynamic markers can be valuable in optimizing liposomal drugs and dosing regimens.

Porphyromonas gingivalis lipopolysaccharide induces interleukin-6 and c-c motif chemokine ligand 2 expression in cultured hCMEC/D3 human brain microvascular endothelial cells.

OBJECTIVE: This paper describes the effect of Porphyromonas gingivalis (P gingivalis) lipopolysaccharide (LPS) on the expression of interleukin-6 (IL-6) and C-C motif chemokine ligand 2 (CCL2) in cultured hCMEC/D3 human brain microvascular endothelial cells. BACKGROUND: P gingivalis is one of the important pathogens in periodontitis, and periodontitis is a risk factor for brain disorders including cerebrovascular diseases and Alzheimer's disease. However, the mechanisms underlying the pathogenesis of P gingivalis-mediated brain diseases are incompletely understood. Effects of P gingivalis LPS on brain endothelial cells are not known well. METHODS: The hCMEC/D3 human brain microvascular endothelial cells were cultured and treated with P gingivalis LPS. The expression of IL-6 and CCL2 mRNA and protein was examined using quantitative reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. Effect of inhibitors of Toll-like receptor (TLR) 2, TLR4, nuclear factor-κB (NF-κB), p38 mitogen-activated protein kinase (MAPK) and c-Jun N-terminal kinase (JNK) was also investigated. Phosphorylation of NF-κB p65, p38 MAPK and JNK was examined using Western blotting. RESULTS: P gingivalis LPS-induced mRNA and protein expression of IL-6 and CCL2 in hCMEC/D3 cells in a concentration-dependent manner at the concentration of 0.5-50 µg/mL. Maximal mRNA expression of IL-6 and CCL2 was found 2 and 4 hours after stimulation, respectively. Induction of IL-6 and CCL2 by P gingivalis LPS was almost completely inhibited by pretreatment of cells with TLR4 inhibitor but not by TLR2 inhibitor. Treatment of cells with P gingivalis LPS for up to 2 hours induced phosphorylation of NF-κB p65, p38 MAPK and JNK. IL-6 induction was decreased by pretreatment of cells with NF-κB inhibitor SN50 or p38 MAPK inhibitor SB203580, while CCL2 induction was reduced by SN50 or JNK inhibitor SP600125. CONCLUSIONS: IL-6 and CCL2 produced upon P gingivalis LPS stimulation may contribute to the inflammatory reactions in brain endothelial cells and subsequent neurological disorders such as cerebrovascular and Alzheimer's diseases.

A Liposomal Gemcitabine, FF-10832, Improves Plasma Stability, Tumor Targeting, and Antitumor Efficacy of Gemcitabine in Pancreatic Cancer Xenograft Models.

PURPOSE: The clinical application of gemcitabine (GEM) is limited by its pharmacokinetic properties. The aim of this study was to characterize the stability in circulating plasma, tumor targeting, and payload release of liposome-encapsulated GEM, FF-10832. METHODS: Antitumor activity was assessed in xenograft mouse models of human pancreatic cancer. The pharmacokinetics of GEM and its active metabolite dFdCTP were also evaluated. RESULTS: In mice with Capan-1 tumors, the dose-normalized areas under the curve (AUCs) after FF-10832 administration in plasma and tumor were 672 and 1047 times higher, respectively, than after using unencapsulated GEM. The tumor-to-bone marrow AUC ratio of dFdCTP was approximately eight times higher after FF-10832 administration than after GEM administration. These results indicated that liposomal encapsulation produced long-term stability in circulating plasma and tumor-selective targeting of GEM. In mice with Capan-1, SUIT-2, and BxPC-3 tumors, FF-10832 had better antitumor activity and tolerability than GEM. Internalization of FF-10832 in tumor-associated macrophages (TAMs) was revealed by flow cytometry and confocal laser scanning microscopy, and GEM was efficiently released from isolated macrophages of mice treated with FF-10832. These results suggest that TAMs are one of the potential reservoirs of GEM in tumors. CONCLUSION: This study found that FF-10832 had favorable pharmacokinetic properties. The liposomal formulation was more effective and tolerable than unencapsulated GEM in mouse xenograft tumor models. Hence, FF-10832 is a promising candidate for the treatment of pancreatic cancer.

Accuracy of an automated method of 3D soft tissue landmark detection.

INTRODUCTION: Due to technological advances, the quantification of facial form can now be done via three-dimensional (3D) photographic systems such as stereophotogrammetry. To enable comparison with traditional cephalometry, soft-tissue anatomical landmark definitions have been modified to incorporate the third dimension. Annotating these landmarks manually, however, is still a time-consuming and arduous process. OBJECTIVE: To develop an automated algorithm to accurately identify anatomical landmarks on three-dimensional soft tissue images. METHODS: Thirty 3dMD images were selected from a private orthodontic practice consisting of 15 males and 15 females between 9 and 17 years of age. The soft-tissue 3D images were aligned along a reference plane to setup a Cartesian coordinate system. Screened by 2 observers, 21 landmarks were manually annotated and their coordinates defined. An automated landmark identification algorithm, based on their anatomical definitions, was developed to compare the landmark validity against the manually identified counterpart. RESULTS: Twenty-one landmarks were analysed in detail. Inter-observer and intra-observer reliability using ICC was >0.9. The average difference and standard deviation between manual and automated methods for all landmarks was 3.2 and 1.64 mm, respectively. Sixteen out of twenty-one landmarks had a mean difference less than 4 mm. The landmarks of greatest agreement (≤2 mm) were mainly in the midline: pronasale, subnasale, subspinale, labiale superius, stomion, with the exception of chelion right. Five linear facial measurements were found to have moderate to good agreement between the manual and automated identification methods. CONCLUSIONS: The developed algorithm was determined to be clinically relevant in the detection of midsagittal landmarks and associated measurements within the studied sample of adolescent Caucasian subjects.

FF-10832 enables long survival via effective gemcitabine accumulation in a lethal murine peritoneal dissemination model.

Chemotherapy has been the treatment of choice for unresectable peritoneal dissemination; however, it is difficult to eradicate such tumors because of poor drug delivery. To solve this issue, we developed FF-10832 as liposome-encapsulated gemcitabine to maintain a high concentration of gemcitabine in peritoneal tumors from the circulation and ascites. A syngeneic mouse model of peritoneal dissemination using murine Colon26 cell line was selected to compare the drug efficacy and pharmacokinetics of FF-10832 with those of gemcitabine. Despite the single intravenous administration, FF-10832 treatment enabled long-term survival of the lethal model mice as compared with those treated with gemcitabine. Pharmacokinetic analysis clarified that FF-10832 could achieve a more effective gemcitabine delivery to peritoneal tumors owing to better stability in the circulation and ascites. The novel liposome-encapsulated gemcitabine FF-10832 may be a curative therapeutic tool for cancer patients with unresectable peritoneal dissemination via the effective delivery of gemcitabine to target tumors.

Vapochromic luminescence and flexibility control of porous coordination polymers by substitution of luminescent multinuclear Cu(I) cluster nodes.

Two luminescent porous coordination polymers (PCPs), i.e., [Cu2(µ2-I)2ctpyz]n and [Cu4(µ3-I)4ctpyz]n (Cu2 and Cu4, respectively; ctpyz = cis-1,3,5-cyclohexanetriyl-2,2',2″-tripyrazine), were successfully synthesized and characterized by single-crystal X-ray diffraction and luminescence spectroscopic measurements. Cu2 consists of rhombus-type dinuclear {Cu2I2} cores bridged by ctpyz ligands, while Cu4 is constructed of cubane-type tetranuclear {Cu4I4} cores bridged by ctpyz ligands. The void fraction of Cu4 is estimated to be 48.0%, which is significantly larger than that of Cu2 (19.9%). Under UV irradiation, both PCPs exhibit red luminescence at room temperature in the solid state (λem values of 660 and 614 nm for Cu2 and Cu4, respectively). Although the phosphorescence of Cu2 does not change upon removal and/or adsorption of EtOH solvent molecules in the porous channels, the solid-state emission maximum of Cu4 red-shifts by 36 nm (λem = 650 nm) upon the removal of the adsorbed benzonitrile (PhCN) molecules from the porous channels (and vice versa). This large difference in the vapochromic behavior of Cu2 and Cu4 is closely related to the framework flexibility. The framework of Cu2 is sufficiently rigid to retain the porous structure without solvated EtOH molecules, whereas the porous structure of Cu4 collapses easily after removal of the adsorbed PhCN molecules to form a nonporous amorphous phase. The original vapor-adsorbed porous structure of Cu4 is regenerated by exposure of the amorphous solid to not only PhCN vapor but also tetrahydrofuran, acetone, ethyl acetate, and N,N-dimethylformamide vapors. The Cu4 structures with the various adsorbed solvents showed almost the same emission maxima as the original PhCN-adsorbed Cu4, except for DMF-adsorbed Cu4, which showed no luminescence probably because of weak coordination of the DMF vapor molecules to the Cu(I) centers of the tetranuclear {Cu4I4} core.

Poor oral health, that is, decreased frequency of tooth brushing, is associated with endothelial dysfunction.

BACKGROUND: Poor oral health is an independent predictor of cardiovascular outcome. Endothelial dysfunction is the initial step of atherosclerosis, resulting in cardiovascular outcomes; but there is no information on the association between oral health and endothelial function. The purpose of this study was to determine the relationships between oral health and endothelial function. METHODS AND RESULTS: A total of 190 subjects who underwent health examinations (mean age, 57±18 years), including patients with cardiovascular disease, completed a questionnaire on oral health and frequency of tooth brushing, and underwent measurement of vascular function, flow-mediated vasodilation (FMD) and nitroglycerine-induced vasodilation. The subjects were divided into 2 groups according to frequency of tooth brushing (≥twice/day and

Allergy to Omalizumab: Lessons from a Reaction to the Coronavirus 2019 Vaccine.

Omalizumab can cause hypersensitivity reactions. We herein report the first case of an 18-year-old woman with refractory cough-predominant asthma that correlated with allergic reactions caused by omalizumab and the coronavirus disease 2019 (COVID-19) vaccine. The patient developed angioedema after taking omalizumab. She had previously experienced intense coughing immediately after receiving a COVID-19 vaccine. A skin prick test was positive for polysorbate 20, which was probably the cause of the allergic reactions to omalizumab and the COVID-19 vaccine. Clinicians should check for an allergic reaction, irrespective of its intensity, triggered by polysorbate and be careful when prescribing biologics to patients in order to avoid allergic reactions.

Model analysis of local oxygen delivery with liposome-encapsulated hemoglobin.

Liposome-encapsulated hemoglobins (LHs) are comparable to red blood cells (RBCs) in terms of oxygen (O(2))-carrying capacity. The smaller particle size of LHs than of platelets allows their homogeneous dispersion in circulating plasma. In this study, we evaluated the effect of LH transfusion on arterial O(2) delivery through vascular trees by simulation. A mathematical model was established on the basis of the coronary arterial anatomy, the conservation of flow and RBC flux, and Poiseuille's law. The Fåhraeus-Lindqvist, Fåhraeus, and phase separation effects were considered in the model. By assuming steady perfusion, the arterial flow and O(2) delivery were calculated for five model trees undergoing the isovolumic replacement of RBCs (0.3 mg hemoglobin (Hb)/mL) with LHs (0.2 mg Hb/mL) or a plasma volume expander (PVE). The RBC-LH exchange increased both the total flow and the total O(2) flux but had almost no effect on the relative distribution of O(2) flux. In contrast, the RBC-PVE exchange decreased the total O(2) flux and increased the proportion of regions receiving a relatively low O(2) supply. Thus, LH transfusion may compensate for an enhanced bias in RBC-associated O(2) flux under hemodilution and is expected to be beneficial for both total and local O(2) delivery.

Drastically progressive lung cavity lesion caused by Actinomyces odontolyticus in a patient undergoing chemoradiotherapy: A case report and literature review.

Pulmonary actinomycosis reportedly forms 15% of all cases of actinomycosis, and pulmonary Actinomyces odontolyticus is particularly rare. A 60-year-old man with a hoarse voice was referred to our hospital. Lung squamous cell carcinoma was diagnosed at the clinical tumor-node-metastasis stage of cT2N2M0, and concurrent chemoradiotherapy was initiated. Further, a small cavity was also detected in the left upper lobe, but it was observed. During chemoradiotherapy, the small cavity lesion rapidly increased accompanying infiltration, and administration of short-term antibiotics did not improve the patient's condition. Bronchoscopy did not show any diagnostic results. Although a rapidly progressive malignant lesion could not be excluded and surgical management was considered, resection could not be performed because of the tight adhesion of the mass. Therefore, bronchoscopy was performed again, and the bronchial lavage culture showed a positive smear for the Actinomyces species. Further, using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), the bacteria was identified as A. odontolyticus. After long-term administration of amoxicillin, the lung cavity with infiltration gradually improved. To the best of our knowledge, there have been nine cases of pulmonary A. odontolyticus (excluding those with only empyema or pleural mass without lung lesions), which can occur in immunocompetent patients with persistent lung shadow. None of the cases showed drastic deterioration; therefore, the present case is the first to highlight that A. odontolyticus possibly produce drastically progressive lung cavity lesion. Further, repeated bronchoscopy and MALDI-TOF MS could help to diagnose pulmonary actinomycosis.

Numerical study on flows of red blood cells with liposome-encapsulated hemoglobin at microvascular bifurcation.

Flow analysis at microvascular bifurcation after partial replacement of red blood cell (RBC) with liposome-encapsulated hemoglobin (LEH) was performed using the lattice Boltzmann method. A two-dimensional symmetric Y bifurcation model with a parent vessel diameter of 20 mum and daughter branch diameters of 20 microm was considered, and the distributions of the RBC, LEH, and oxygen fluxes were calculated. When only RBCs flow into the daughter branches with unevenly distributed flows, plasma separation occurred and the RBC flow to the lower-flow branch was disproportionately decreased. On the other hand, when half of RBC are replaced by LEH, the biasing of RBC flow was enhanced whereas LEH flowed favorably into the lower-flow branch, because many LEH within the parent vessel are suspended in the plasma layer, where no RBCs exist. Consequently, the branched oxygen fluxes became nearly proportional to flows. These results indicate that LEH facilitates oxygen supply to branches that are inaccessible to RBCs.

Myocardial microvascular perfusion after transfusion of liposome-encapsulated hemoglobin evaluated in cross-circulated rat hearts using tracer digital radiography.

The effect of hemodilution with Neo Red Cell (NRC, liposome-encapsulated hemoglobin) on myocardial perfusion was evaluated in cross-circulated rat hearts under 300-bpm pacing and 100-mmHg perfusion pressure. In NRC-transfused hearts (n = 5), NRC volume fraction and hematocrit were 9% +/- 3% and 22% +/- 4%, respectively; the latter decreased from 43% +/- 3% before NRC transfusion. Coronary perfusion rate and left ventricular isovolemic developed pressure increased after NRC transfusion to 4.6 +/- 1.0 ml/min/g and 127 +/- 32 mmHg from basal values of 2.5 +/- 0.3 ml/min/g and 115 +/- 28 mmHg, respectively. In contrast, the flow increase during reperfusion following 30-s flow cessation decreased from 74% +/- 24% to 64% +/- 24%. The arteriovenous difference in O2 saturation was slightly higher after NRC transfusion. Within-layer regional flow distributions from subepicardium to subendocardium assessed by tracer digital radiography (100-microm resolution) showed that coefficients of variation of flows in 400 x 400-microm regions were 0.41 +/- 0.10 in NRC-transfused hearts and 0.54 +/- 0.11 in nontransfused hearts (n = 5); i.e., the myocardial flow distribution was more uniform in NRC-transfused hearts. These results suggest that NRC is superior to erythrocytes in terms of the homogenization of O2 delivery, indicating its potential therapeutic value in myocardial microcirculatory failure.

Reabilitação parcial sobreimplantes. A prótese fixa destacável / Partial rehabilitation on osseointegrated implants. The fixed-detachable prosthesis

O implante dentário já é uma realidade do dia-a-dia do consultório, ampliando as possibilidades de reabilitações mais eficientes. Entretanto, mesmo com os implantes, o dilema consiste em oferecer uma solução que atenda o paciente e ao mesmo tempo atenda aos requisitos fundamentais de higiene e manutenção sobre o qual toda viabilidade do implante está fundamentada. O presente relato sugere uma das alternativas para esta realização, mantendo uma proposta que não perde o foco estético, mas prioriza a manutenção simples, bem como o custo de produção razoável, tornando esta proposta socialmente abrangente.