A tetravalent dengue virus-like particle vaccine induces high levels of neutralizing antibodies and reduces dengue replication in non-human primates.

Dengue virus (DENV) represents a significant global health burden, with 50% of the world's population at risk of infection, and there is an urgent need for next-generation vaccines. Virus-like particle (VLP)-based vaccines, which mimic the antigenic structure of the virus but lack the viral genome, are an attractive approach. Here, we describe a dengue VLP (DENVLP) vaccine which generates a neutralizing antibody response against all four DENV serotypes in 100% of immunized non-human primates for up to 1 year. Additionally, DENVLP vaccination produced no ADE response against any of four DENV serotypes in vitro. DENVLP vaccination reduces viral replication in a non-human primate challenge model. We also show that transfer of purified IgG from immunized monkeys into immunodeficient mice protects against subsequent lethal DENV challenge, indicating a humoral mechanism of protection. These results indicate that this DENVLP vaccine is immunogenic and can be considered for clinical evaluation. Immunization of non-human primates with a tetravalent DENVLP vaccine induces high levels of neutralizing antibodies and reduces the severity of infection for all four dengue serotypes.IMPORTANCEDengue is a viral disease that infects nearly 400 million people worldwide and causes dengue hemorrhagic fever, which is responsible for 10,000 deaths each year. Currently, there is no therapeutic drug licensed to treat dengue infection, which makes the development of an effective vaccine essential. Virus-like particles (VLPs) are a safe and highly immunogenic platform that can be used in young children, immunocompromised individuals, as well as healthy adults. In this study, we describe the development of a dengue VLP vaccine and demonstrate that it induces a robust immune response against the dengue virus for over 1 year in monkeys. The immunity induced by this vaccine reduced live dengue infection in both murine and non-human primate models. These results indicate that our dengue VLP vaccine is a promising vaccine candidate.

Relationship Between Submental Superficial Layer Hardness and Tongue Pressure Determined Using a Tissue Durometer.

Aging affects the tongue and suprahyoid muscles, causing dysphagia and undernutrition. We hypothesized that tongue function would affect submental superficial layer hardness. Tongue movements during water retention between the tongue and palate are the same as those required during bolus formation, involving internal and external tongue muscle movement. In 28 healthy adults (14 males, 14 females, average age 28.7 ± 2.9 years), we measured the submental superficial layer characteristics (frequency [tension], stiffness, decrement [inverse of resilience], relaxation, and creep [deformation over time]) using a simple tissue durometer (MyotonPRO®), and examined their relationship with maximum voluntary tongue pressure. The tissue durometer sensor was placed in the submental region, where there is no intervening bone. Measurements were performed at rest and while retaining 5 mL water. Tongue pressure was measured using a tongue pressure-measuring device. The submental superficial layer hardness differed significantly between rest and during water retention. During water retention, frequency and stiffness were high, while decrement, relaxation, and creep were low. When pressure is applied to the palate, such as during water retention the inner tongue muscle, which changes the tongue's shape, and the outer tongue muscle, which moves the tongue laterally, are active. However, the change in the hardness of the submental superficial layer during water retention may be related to the suprahyoid muscles that are present in this layer. The results of this study suggested that the hardness of the submental superficial layer changed during water retention, and that tongue movement could be measured from outside the mouth.

Direct enantioseparation of mandelic acid by high-performance liquid chromatography using a phenyl column precoated with a small amount of cyclodextrin additive in a mobile phase.

Direct enantioseparation of mandelic acid by high-performance liquid chromatography (HPLC) with a reversed phase column and a mobile phase containing a small amount of hydroxylpropyl-ß-cyclodextrin (HP-ß-CD) was studied as an efficient method for saving consumption of the CD additive. As a result, it was proposed that racemic mandelic acid can be analyzed with a phenyl column by using a mobile phase composed of 10 mM ammonium acetate buffer (pH 4.2) and 0.02% (w/v) HP-ß-CD at a flow rate of 1.0 mL/min at 40°C after the passage of 10 mM ammonium acetate buffer (pH 4.2) containing 0.1% (w/v) HP-ß-CD as a precoating mobile phase for 60 min. It is suggested that HP-ß-CD is bound with a phenyl group on the surface of the stationary phase to allow a phenyl column to act as a transient chiral column, and injected mandelic acid can form the ternary complex with the adsorbed HP-ß-CD. The longer retention time of D-mandelic acid than the L-isomer for HPLC can be explained from the higher stability of the HP-ß-CD complex with D-mandelic acid, which was confirmed by CE experiment with HP-ß-CD as a selector. The efficiency of a phenyl column compared with other stationary phases was also discussed.

Evaluation of the Newly Developed Adenosine Diphosphate-Induced Platelet Aggregation Level System in Aggregometer on Automated Coagulation Analyzer.

BACKGROUND: Light transmission aggregometry (LTA) is the gold standard for platelet function assessment. The automated coagulation analyzer from Sysmex that performs LTA offers the advantage of being a walk-away technology. Recently, a new parameter "ADP-induced platelet aggregation level (APAL)" was developed to support the interpretation of results. APAL is calculated as a score from 0.0 to 10.0 based on platelet aggregation patterns with 1 and 10 µM adenosine diphosphate (ADP). Here, the basic performance of the newly developed APAL system and comparison with the maximum aggregation rate of ADP (ADP-MA) was evaluated. METHODS: The within-run precision was calculated by conducting five replicate analyses of the platelet-rich plasma (PRP) from healthy volunteers and 0.05 µM of cangrelor-spiked PRP. Cangrelor is a P2Y12 inhibitor that does not require liver CYP activation. The reference interval was calculated from the results of 67 healthy volunteers. The effect of the antiplatelet P2Y12 agent was evaluated using several concentrations of cangrelor. A comparative study was performed using 103 PRP samples with different levels of aggregation. Each test was analyzed with both APAL and ADP-MA. RESULTS: The percentage coefficient of variation in within-run precision was within 7% for APAL and 10 µM ADP-MA. Reference interval of APAL and 10 µM ADP-MA was 7.1 - 10.0 and 80.0 - 99.2%, respectively. APAL signifi-cantly decreased with the addition of 0.02 µM cangrelor, while 10 µM ADP-MA was barely affected. A significant correlation was observed between APAL and 10 µM ADP-MA (r = 0.94; p < 0.0001). CONCLUSIONS: The newly developed APAL system exhibited an acceptable performance. APAL score showed a good correlation with ADP-MA and was adequate to detect the weak effect of P2Y12 inhibitors. APAL is a new platelet aggregation scoring system with the potential to monitor the effects of P2Y12 inhibitor over a wide range.

Membrane transport mechanisms of choline in human intestinal epithelial LS180 cells.

The aim of the present study was to investigate the membrane transport mechanisms of choline using human intestinal epithelial LS180 cells. The mRNA of choline transporter-like proteins (CTLs) was expressed significantly in LS180 cells, and the rank order was CTL1 > CTL4 > CTL3 > CTL2 > CTL5. In contrast, the mRNA expression of other choline transporters, organic cation transporter (OCT) 1, OCT2 and high-affinity choline transporter 1 (CHT1), was considerably lower in LS180 cells. Five mm unlabelled choline, hemicolinium-3 and guanidine, but not tetraethylammonium, inhibited the cellular uptake of 100 µm choline in LS180 cells. The uptake of choline into LS180 cells was virtually Na(+)-independent. The uptake of choline was significantly decreased by acidification of the extracellular pH; however, it was not increased by alkalization of the extracellular pH. In addition, both acidification and alkalization of intracellular pH decreased the uptake of choline, indicating that the choline uptake in LS180 cells is not stimulated by the outward H(+) gradient. On the other hand, the uptake of choline was decreased by membrane depolarization along with increasing extracellular K(+) concentration. In addition, the Na(+)-independent uptake of choline was saturable, and the Km value was estimated to be 108 µm. These findings suggest that the uptake of choline into LS180 cells is membrane potential-dependent, but not outward H(+) gradient-dependent.

Relationship between Knowledge and Attitude toward Oral Function in Middle-Aged and Older Adults.

Oral hypofunction is the stage at which recovery can be expected with proper diagnosis, management, and motivation before oral dysfunction occurs. The knowledge and attitude toward oral function can influence the maintenance and improvement of oral function. However, whether middle-aged and older adults with declining oral function have knowledge of their oral function and how this knowledge and their attitude affect their oral function are unclear. Therefore, we aimed to examine (1) the relationship between knowledge and attitude toward oral function and hypofunction in individuals with suspected oral hypofunction and (2) changes in knowledge and attitude toward oral function through evaluation and education. Participants aged ≥50 years were enrolled during their first community dental clinic visit. A questionnaire assessment of knowledge and attitudes before and after oral function evaluation was performed. The oral function was initially assessed with seven criteria: oral hygiene; oral dryness; occlusal force; tongue pressure; tongue-lip motor, masticatory, and swallowing function. Associations between knowledge and attitudes and their changes were statistically analyzed. Fifty-nine participants (93.7%) were unaware of "oral hypofunction." Associations between knowledge and attitudes and their changes in the negative to positive response groups, from 86.4% and 61.0% to 6.8% and 25.4%, respectively, after oral function evaluation, indicated that participants understood their oral function and the need for training. Middle-aged and older individuals with poor knowledge and attitudes were more likely to have a worse oral function; however, their knowledge and attitudes toward oral function could be improved through oral function assessment and education.

Antigen-specific CD4+ effector T cells: analysis of factors regulating clonal expansion and cytokine production: clonal expansion and cytokine production by CD4+ effector T cells.

In order to fully understand T cell-mediated immunity, the mechanisms that regulate clonal expansion and cytokine production by CD4(+) antigen-specific effector T cells in response to a wide range of antigenic stimulation needs clarification. For this purpose, panels of antigen-specific CD4(+) T cell clones with different thresholds for antigen-induced proliferation were generated by repeated stimulation with high- or low-dose antigen. Differences in antigen sensitivities did not correlate with expression of TCR, CD4, adhesion or costimulatory molecules. There was no significant difference in antigen-dependent cytokine production by TG40 cells transfected with TCR obtained from either high- or low-dose-responding T cell clones, suggesting that the affinity of TCRs for their ligands is not primary determinant of T cell antigen reactivity. The proliferative responses of all T cell clones to both peptide stimulation and to TCRbeta crosslinking revealed parallel dose-response curves. These results suggest that the TCR signal strength of effector T cells and threshold of antigen reactivity is determined by an intrinsic property, such as the TCR signalosome and/or intracellular signaling machinery. Finally, the antigen responses of high- and low-peptide-responding T cell clones reveal that clonal expansion and cytokine production of effector T cells occur independently of antigen concentration. Based on these results, the mechanisms underlying selection of high "avidity" effector and memory T cells in response to pathogen are discussed.

[Report of a case effectively treated by appropriate therapy for severe anticipatory nausea and vomiting due to FOLFOX or FOLFIRI].

To date, the standard treatments for severe anticipatory nausea and vomiting is not well established. 5-HT3 antagonist is one of the effective drugs to reduce chemotherapy-induced nausea and vomiting, but had no effect on these symptoms for this patient. The patient could be successfully administered standard chemotherapy(FOLFOX or FOLFIRI, q2w)without adverse reactions by appropriate treatments in the form of increased doses of dexamethasone and normal dose administration of prochlorperazine. This report suggests a possibility that FOLFOX or FOLFIRI may be successfully treated by appropriate treatments for severe chemotherapy-induced vomiting colon cancer patients, and that this observation may lead to the improved prognosis of these patients.

[A case of left urothelial carcinoma with multiple bone metastasis, liver metastasis and right adrenal metastasis successfully treated with combination chemotherapy of gemcitabine and carboplatin].

The patient was a 53-year-old male. He had been admitted to another hospital with a complaint of left sciatica. He was referred to our hospital for further examination and therapy. He was diagnosed as left urothelial carcinoma with multiple bone metastasis, liver metastasis and right adrenal metastasis. He was treated with combination chemotherapy of gemcitabine and carboplatin (1,000 mg/m2 day 1 and AUC 2 day 1, respectively) biweekly. After the ninth course, a significant tumor reduction was obtained, and has been maintained. He has been treated on an outpatient basis because of no grade 3 or severer adverse reactions. We report an effective case of biweekly chemotherapy with gemcitabine and carboplatin in the treatment of advanced urothelial carcinoma.

Improvement of regional cerebral blood flow after treatment with branched-chain amino acid solutions in patients with cirrhosis.

OBJECTIVE: The administration of solutions rich in branched-chain amino acids leads to mental recovery from acute hepatic encephalopathy in patients with liver cirrhosis. However, the mechanism of action of branched-chain amino acids remains unclear. The purpose of this study was to evaluate the effect of intravenous infusion of branched-chain amino acids on brain perfusion in patients with liver cirrhosis. METHODS: Single photon emission computed tomography scans were performed in 14 patients with liver cirrhosis before and after the administration of branched-chain amino acids in a single-day split-dose protocol. The per cent change in regional brain perfusion was calculated in high frontal, parietal, temporal, occipital lobes and cerebellum. Thereafter, statistical parametric mapping was performed to identify brain regions with abnormal cerebral perfusion. RESULTS: Intravenous infusion of solutions enriched with branched-chain amino acids induced a 13-20% increase in regional cerebral blood flow. Cirrhotic patients had regions of significant hypoperfusion, as determined by statistical parametric mapping, in the left superior parietal and posterior cingulate as compared to the control group. This hypoperfusion of parietal and cingulate regions was not detected after treatment with solutions of branched-chain amino acids. CONCLUSIONS: The results of the present study suggest that administration of solutions enriched with branched-chain amino acids improves cerebral perfusion in patients with cirrhosis.

FOLFIRI Is Tolerable after Subtotal Colectomy - A Patient with Familial Adenomatous Polyposis Who Developed Advanced Rectal Cancer.

A 40-year-old female with familial adenomatous polyposis (FAP) had a subtotal colectomy at 16 years of age. At 39 years, she had low anterior resection due to advanced rectal carcinoma. Thereafter, we administrated per os uracil and tegafur for 9 months. Metastatic rectal carcinoma was detected in the liver (S8) by computed tomography (CT). 2-[(18)F]-fluoro-2-deoxy-D-glucose (FDG) positron emission tomography (PET) data did not show any other metastasis. This report presents a first case of a patient undergoing subtotal colectomy administered FOLFIRI (CPT-11 180 mg/m(2) as a 90-minute infusion on day 1; leucovorin 400 mg/m(2) as a 2-hour infusion during CPT-11, immediately followed by 5-FU bolus 400 mg/m(2) and 46-hour continuous infusion of 2,400 mg/m(2) every 2 weeks). This regimen was administered without grade 3 or 4 of any adverse reaction for 6 months, although there was a possibility that this patient with subtotal colectomy may have the cause for severe diarrhea. Further investigations are needed to assess the safety in clinical trials of FOLFIRI regimen for patients with subtotal colectomy.

Anaplastic transformation from papillary thyroid carcinoma with increased serum CA19-9.

A 24-year-old woman presented with anaplastic transformation from papillary thyroid carcinoma with increased serum CA19-9. The patient had been diagnosed as having papillary thyroid carcinoma with lung metastasis at 11 years of age. She received a total thyroidectomy with cervical lymph node dissection followed by iodine-131 ((131)I) therapy over 12 years, but died due to sudden onset of rapid dissemination. Elevated serum CA19-9 was detected in the terminal stage, and anaplastic transformation was confirmed by post-mortem examination. Although there are few clinical reports suggesting a prognostic indicator for anaplastic thyroid carcinoma, CA19-9 may be a useful serum marker for this tumor.

Lymphatic mapping and sentinel node biopsy during laparoscopic gastrectomy for early cancer.

BACKGROUND/AIMS: Intraoperative lymphatic mapping and sentinel node (SN) biopsy can potentially be combined with minimally invasive surgery, but there are few reports of laparoscopic lymphatic mapping for gastrointestinal cancer. We examined the feasibility and accuracy of laparoscopic lymphatic mapping in predicting lymph node status in patients with gastric cancer. METHODS: Seventeen patients with gastric cancer invading the mucosal or submucosal layers (T1) underwent laparoscopic gastrectomy with lymphatic mapping between March 2001 and May 2002. The day before surgery, a technetium-99m-labelled tin colloid solution was injected endoscopically around the tumor. Immediately after the pneumoperitoneum, patent blue was injected. Gastrectomy was performed in all patients, and blue-stained or radioactive nodes were defined as SNs. Fresh SNs were immediately processed for frozen-section examination by hematoxylin-eosin (H and E) and immunohistochemical (IHC) staining. All non-SNs harvested from resected specimens were subjected to histological examination with H and E. RESULTS: SNs were detected in all patients by combination of the two kinds of tracers. Three patients had lymph node metastases in their final examination, and SNs in these 3 were operatively diagnosed as positive by H and E or IHC staining. Lymphatic mapping and SN biopsy under laparoscopic surgery were performed with 100% accuracy. CONCLUSION: Our preliminary study shows the feasibility of intraoperative lymphatic mapping in laparoscopic gastrectomy for T1 gastric cancer.