Protective efficacy of an IL-12-expressing baculoviral malaria vaccine.

Interleukin-12 (IL-12) plays an important role in antigen-specific adaptive immunity against Plasmodium sporozoites, and this requirement allows for a new approach to developing an effective malaria vaccine. In this study, we examined whether IL-12 could enhance protective efficacy of a baculovirus-based malaria vaccine. For this aim, a baculoviral vector expressing murine IL-12 (mIL-12) under the control of CMV promoter (BES-mIL-12-Spider) and a baculoviral vector expressing Plasmodium falciparum circumsporozoite protein (PfCSP) with post-transcriptional regulatory element of woodchuck hepatitis virus (BDES-sPfCSP2-WPRE-Spider) were generated. BES-mIL-12-Spider produced bioactive IL-12 which activates splenocytes, resulting in induction of IFN-γ. When co-immunized with BES-mIL-12-Spider and BDES-sPfCSP2-WPRE-Spider, the mouse number for high IgG2a/IgG1 ratios and the geometric mean in this group were both increased as compared with those of the other groups, indicating a shift towards a Th1-type response following immunization with BES-mIL-12-Spider. Finally, immunization with BDES-sPfCSP2-WPRE-Spider plus BES-mIL-12-Spider had a higher protective efficacy (73%) than immunization with BDES-sPfCSP2-WPRE-Spider alone (30%) against challenge with transgenic Plasmodium berghei sporozoites expressing PfCSP. These results suggest that co-administration of IL-12 expressing baculoviral vector, instead of IL-12 cDNA, with viral-vectored vaccines provides a new feasible vaccine platform to enhance Th1-type cellular immune responses against Plasmodium parasites.

Prognostic nomogram for nonresectable pancreatic cancer treated with gemcitabine-based chemotherapy.

BACKGROUND: A nomogram is progressively being used as a useful predictive tool for cancer prognosis. A nomogram to predict survival in nonresectable pancreatic cancer treated with chemotherapy has not been reported. METHODS: Using prospectively collected data on patients with nonresectable pancreatic cancer receiving gemcitabine-based chemotherapy at five Japanese hospitals, we derived a predictive nomogram and internally validated it using a concordance index and calibration plots. RESULTS: In total, 531 patients were included between June 2001 and February 2013. The American Joint Committee on Cancer (AJCC) TNM stages were III and IV in 204 and 327 patients, respectively. The median survival time of the total cohort was 11.3 months. A nomogram was generated to predict survival probabilities at 6, 12, and 18 months and median survival time, based on the following six variables: age; sex; performance status; tumour size; regional lymph node metastasis; and distant metastasis. The concordance index of the present nomogram was higher than that of the AJCC TNM staging system at 12 months (0.686 vs 0.612). The calibration plots demonstrated good fitness of the nomogram for survival prediction. CONCLUSIONS: The present nomogram can provide valuable information for tailored decision-making early after the diagnosis of nonresectable pancreatic cancer.

Insulin stimulates the expression of the SHARP-1 gene via multiple signaling pathways.

The rat enhancer of split- and hairy-related protein-1 (SHARP-1) is a basic helix-loop-helix transcription factor. An issue of whether SHARP-1 is an insulin-inducible transcription factor was examined. Insulin rapidly increased the level of SHARP-1 mRNA both in vivo and in vitro. Then, signaling pathways involved with the increase of SHARP-1 mRNA by insulin were determined in H4IIE rat hepatoma cells. Pretreatments with LY294002, wortmannin, and staurosporine completely blocked the induction effect, suggesting the involvement of both phosphoinositide 3-kinase (PI 3-K) and protein kinase C (PKC) pathways. In fact, overexpression of a dominant negative form of atypical protein kinase C lambda (aPKCλ) significantly decreased the induction of the SHARP-1 mRNA. In addition, inhibitors for the small GTPase Rac or Jun N-terminal kinase (JNK) also blocked the induction of SHARP-1 mRNA by insulin. Overexpression of a dominant negative form of Rac1 prevented the activation by insulin. Furthermore, actinomycin D and cycloheximide completely blocked the induction of SHARP-1 mRNA by insulin. Finally, when a SHARP-1 expression plasmid was transiently transfected with various reporter plasmids into H4IIE cells, the promoter activity of PEPCK reporter plasmid was specifically decreased. Thus, we conclude that insulin induces the SHARP-1 gene expression at the transcription level via a both PI 3-K/aPKCλ/JNK- and a PI 3-K/Rac/JNK-signaling pathway; protein synthesis is required for this induction; and that SHARP-1 is a potential repressor of the PEPCK gene expression.

Radiocaesium concentrations in the muscle and eggs of salmonids from Lake Chuzenji, Japan, after the Fukushima fallout.

Approximately 18 months (September to December 2012) after the Fukushima Dai-ichi Nuclear Power Plant accident, elevated radiocaesium concentrations were measured in samples of muscle and eggs from masu salmon Oncorhynchus masou, kokanee Oncorhynchus nerka, brown trout Salmo trutta and lake trout Salvelinus namaycush from the Lake Chuzenji system, central Honshu Island, Japan (160 km from the station). Mean muscle concentrations were 142·9-249·2 Bq kg⁻¹ wet mass and mean concentrations in eggs were 38·7-79·0 Bq kg⁻¹ wet mass. There was no relationship between fork length and muscle radiocaesium concentration in any of the species, but there were significant relationships between individual muscle and egg radiocaesium concentrations from O. masou, S. trutta and S. namaycush.

TbCu7-type Sm-Fe(-N) powder synthesized by low-temperature reduction-diffusion process using the Li-Ca reductant.

It was predicted that TbCu7-type Sm-Fe powder prepared by the low-temperature reduction-diffusion (LTRD) process using a Li-Ca reductant would contain no residual ɑ-Fe because this reductant would not produce the absorbed water that hinders the reaction between Sm and Fe by forming oxychlorides when molten salt is used as the reductant. Contrary to this expectation, a detailed microstructure analysis revealed that a residual phase of unreacted ɑ-Fe existed in some TbCu7-type Sm-Fe particles rather than as separate Fe particles. This residual ɑ-Fe phase was not located in the center of the Sm-Fe particles and was not detected in some Sm-Fe particles, suggesting that the reason for the residual ɑ-Fe phase is inhomogeneous diffusion of Sm into the Fe due to slow diffusion at low temperatures. Although this TbCu7-type Sm-Fe powder contained a small amount of unreacted ɑ-Fe phase, the magnetic properties of the nitride TbCu7-type Sm-Fe were also estimated.

Combination of platelet count and neutrophil to lymphocyte ratio is a useful predictor of postoperative survival in patients with colorectal cancer.

BACKGROUND: This study investigated the usefulness of a novel inflammation-based prognostic system, named the COP-NLR (COmbination of Platelet count and Neutrophil to Lymphocyte Ratio), for predicting the postoperative survival of patients with colorectal cancer (CRC). METHODS: The COP-NLR was calculated on the basis of data obtained on the day of admission: patients with both an elevated platelet count (>30 × 10(4) mm(-3)) and an elevated NLR (>3) were allocated a score of 2, and patients showing one or neither were allocated a score of 1 or 0, respectively. RESULTS: Four-hundred and eighty patients were enrolled. Multivariate analysis of clinical characteristics selected by univariate analysis showed that the COP-NLR (1, 2/0) (odds ratio, 0.464; 95% confidence interval, 0.267-0.807; P=0.007) had an association with cancer-specific survival, along with pathology, lymph node metastasis, the serum levels of carcinoembryonic antigen, C-reactive protein and albumin, and the Glasgow Prognostic Score. Kaplan-Meier analysis and log-rank test revealed that the COP-NLR was able to divide such patients into three independent groups (P<0.001). CONCLUSION: The COP-NLR is considered to be a useful predictor of postoperative survival in patients with CRC.

Intratumoral concentration of estrogens and clinicopathological changes in ductal carcinoma in situ following aromatase inhibitor letrozole treatment.

BACKGROUND: Estrogens have important roles in ductal carcinoma in situ (DCIS) of the breast. However, the significance of presurgical aromatase inhibitor treatment remains unclear. Therefore, we examined intratumoral concentration of estrogens and changes of clinicopathological factors in DCIS after letrozole treatment. METHODS: Ten cases of postmenopausal oestrogen receptor (ER)-positive DCIS were examined. They received oral letrozole before the surgery, and the tumour size was evaluated by ultrasonography. Surgical specimens and corresponding biopsy samples were used for immunohistochemistry. Snap-frozen specimens were also available in a subset of cases, and used for hormone assays and microarray analysis. RESULTS: Intratumoral oestrogen levels were significantly lower in DCIS treated with letrozole compared with that in those without the therapy. A great majority of oestrogen-induced genes showed low expression levels in DCIS treated with letrozole by microarray analysis. Moreover, letrozole treatment reduced the greatest dimension of DCIS, and significantly decreased Ki-67 and progesterone receptor immunoreactivity in DCIS tissues. CONCLUSION: These results suggest that estrogens are mainly produced by aromatase in DCIS tissues, and aromatase inhibitors potently inhibit oestrogen actions in postmenopausal ER-positive DCIS through rapid deprivation of intratumoral estrogens.

Pathophysiological analysis of uninephrectomized db/db mice as a model of severe diabetic kidney disease.

Diabetic nephropathy, included in diabetic kidney disease (DKD), is the primary disease leading to end-stage renal disease (ESRD) or dialysis treatment, accounting for more than 40% of all patients with ESRD or receiving dialysis. Developing new therapeutics to prevent the transition to ESRD or dialysis treatment requires an understanding of the pathophysiology of DKD and an appropriate animal model for drug efficacy studies. In this study, we investigated the pathophysiology of diabetic kidney disease with type 2 diabetes in uninephrectomized db/db mice. In addition, the nephrectomized db /db mice from 10 weeks to 42 weeks were used to assess the efficacy of long-term administration of the angiotensin-II-receptor antagonist losartan. The blood and urinary biochemical parameters, main pharmacological endpoint of the losartan therapy, were periodically measured. And at the end, histopathological analysis was performed. Uninephrectomized db/db mice clearly developed obesity and hyperglycemia from young age. Furthermore, they showed renal pathophysiological changes, such as increased urinary albumin-creatinine ratio (UACR) (the peak value 3104 ± 986 in 40-week-old mice), glomerular hypertrophy and increased fibrotic areas in the tubulointerstitial tubules. The blood pressure in the losartan group was significantly low compared to the normotensive Vehicle group. However, as expected, Losartan suppressed the increase in UACR (829±500) indicating the medication was sufficient, but the histopathological abnormalities including tubular interstitial fibrosis did not improve. These results suggest that the uninephrectomized db/db mice are useful as an animal model of the severe DKD indicated by the comparison of the efficacy of losartan in this model with the efficacy of losartan in clinical practice.

Cartilaginous repair of full-thickness articular cartilage defects is induced by the intermittent activation of PTH/PTHrP signaling.

OBJECTIVE: We studied the effects of the transient activation of parathyroid hormone (PTH)/PTH-related peptide (PTHrP) signaling during the repair of 5-mm-diameter full-thickness defects of articular cartilage in the rabbit. MATERIALS AND METHODS: Cylindrical full-thickness articular cartilage defects of 5mm in diameter were artificially created in the femoral trochlea of male adolescent Japanese white rabbits using a hand-drill. Recombinant human PTH(1-84) was then administered into the joint cavity continuously or intermittently for 2 weeks post-injury. The reparative tissues were histologically examined at 2, 4, and 8 weeks, and were also immunohistochemically examined for type II collagen. Double immunostaining analysis was also performed for the PTH/PTHrP receptor and proliferating cell nuclear antigen (PCNA) in the regenerating tissues. RESULTS: No evidence of cartilage formation was evident throughout the period of the experiments in injured animals administered saline alone. In contrast, cartilage formation occurred at 4 weeks in both the continuous and intermittent PTH-treated defects. At 8 weeks post-injury, for the intermittently treated defects, the regenerated cartilage successfully resurfaced the defects and the original bone-articular cartilage junction was recovered. In contrast, the defects were covered with fibrous or fibrocartilaginous tissues in the continuously administered group. PCNA and PTH/PTHrP receptor-double positive mesenchymal cells were significantly increased in both the continuous and intermittent PTH-treated defects at 2 weeks post-injury. CONCLUSIONS: The present results suggest that the transient activation and release from PTH/PTHrP signaling during the early stages of the cartilage repair process facilitates the induction of regenerative chondrogenesis in full-thickness articular cartilage defects.

Insufficient lymph node dissection is an independent risk factor for postoperative cancer death in patients undergoing surgery for stage II colorectal cancer.

BACKGROUND: Surgical resection is the most effective treatment for patients with stage II colorectal cancer (CRC). However, a few patients with early-phase CRC suffer postoperative cancer death. AIMS: To investigate the risk factors for postoperative cancer death in patients who undergo surgery for stage II CRC. METHODS: Prognostic significance was analyzed by χ(2) test, univariate analyses, Kaplan-Meier analysis and log-rank test using clinicopathological factors from the database. RESULTS: A total of 205 patients with stage II CRC were evaluated. Age (≤ 65/>65 years), venous invasion (negative/positive), number of dissected lymph nodes (≤ 9/≥ 10), grade of lymph node dissection (sufficient/insufficient) and albumin level (< 3.5/≥ 3.5 g/dl) were associated with shorter overall survival. Univariate analysis of the clinicopathological factors revealed that only the grade of lymph node dissection (sufficient/insufficient) was associated with postoperative cancer death (odds ratio 2.993, 95% confidence interval 1.216-7.368, p = 0.017).Kaplan-Meier analysis and log-rank test revealed that the group with insufficient lymph node dissection had a higher incidence of postoperative cancer death than the group with sufficient dissection (p < 0.001). CONCLUSIONS: Insufficient lymph node dissection is an independent risk factor for postoperative cancer death in patients who undergo surgery for stage II CRC.

Metal ion and vitamin adsorption profiles of phosphate binder ion-exchange resins.

AIMS: To determine the metal ion and vitamin in vitro adsorption profile of sevelamer hydrochloride (sevelamer-HCl) and colestilan(INN)/colestimide(JAN), a novel ion-exchange resin being developed as a phosphate binder for end-stage renal disease (ESRD) patients undergoing hemodialysis, adsorption of metal ions (iron, cobalt, copper and zinc) and vitamins (B6, B12, C, K and folic acid) essential for hematopoiesis/blood coagulation was assessed. METHODS: Mixtures of each resin (colestilan or sevelamer-HCl, 4 mg/ml) and metal ions (Fe(III), Fe(II), Co(II), Cu(II), and Zn(II), 1 microg/ml) were adjusted to pH 1.2 or 6.8 and incubated at 37 degrees C for 1 hour. Metal ions in the recovered filtrate were detected by inductively coupled plasma optical emission spectrometry. In addition, the mixtures of each resin (4 mg/ml) and vitamins (B6, B12, C, K and folic acid, 0.5 - 250 microg/ml) were adjusted to pH 6.8 and incubated at 37 degrees C for 0.5 hour. The vitamin concentrations in the recovered filtrate were quantified by HPLC. RESULTS: Colestilan did not adsorb any metals tested at either pH level, whereas sevelamer-HCl adsorbed copper(II) and zinc(II) ion at pH 6.8 with adsorption ratios of 99% and 38%, respectively. Both resins showed almost complete adsorption of vitamin C, vitamin K, and folic acid, but weak adsorption of vitamin B6, and no adsorption of vitamin B12. CONCLUSIONS: The differing adsorption profiles for metal ions and vitamins between sevelamer-HCl and colestilan may be of importance for the individualized management of anemia and malnutrition in chronic hemodialysis patients receiving phosphate binding ion-exchange resins for the control of hyperphosphatemia.

Human lung microvascular endothelial cells as potential alternatives to human umbilical vein endothelial cells in bio-3D-printed trachea-like structures.

BACKGROUND: We have been trying to produce scaffold-free structures for airway regeneration using a bio-3D-printer with spheroids, to avoid scaffold-associated risks such as infection. Previous studies have shown that human umbilical vein endothelial cells (HUVECs) play an important role in such structures, but HUVECs cannot be isolated from adult humans. The aim of this study was to identify alternatives to HUVECs for use in scaffold-free structures. METHODS: Three types of structure were compared, made of chondrocytes and mesenchymal stem cells with HUVECs, human lung microvascular endothelial cells (HMVEC-Ls), and induced pluripotent stem cell (iPSC)-derived endothelial cells. RESULTS: No significant difference in tensile strength was observed between the three groups. Histologically, some small capillary-like tube formations comprising CD31-positive cells were observed in all groups. The number and diameters of such formations were significantly lower in the iPSC-derived endothelial cell group than in other groups. Glycosaminoglycan content was significantly lower in the iPSC-derived endothelial cell group than in the HUVEC group, while no significant difference was observed between the HUVEC and HMVEC-L groups. CONCLUSIONS: HMVEC-Ls can replace HUVECs as a cell source for scaffold-free trachea-like structures. However, some limitations were associated with iPSC-derived endothelial cells.

Comparison of 0.05% chlorhexidine and 10% povidone-iodine as cutaneous disinfectant for prevention of central venous catheter-related bloodstream infection: a comparative study.

BACKGROUND: The Centers for Disease Control and Prevention guideline recommended the use of 2% chlorhexidine as a percutaneous disinfectant for central venous catheter (CVC) insertion. However, in Japan, 0.05% chlorhexidine is commonly used as well as 10% povidone-iodine, instead of 2% chlorhexidine. PURPOSE: It was the aim of this study to examine whether the use of 0.05% chlorhexidine is inferior to conventional 10% povidone-iodine as a percutaneous disinfectant for preventing CVC-related bloodstream infection (CVC-RBSI). METHODS: Between September 2006 and July 2008, the time interval from insertion to development of CVC-RBSI was compared prospectively between patients prepared with 0.05% chlorhexidine (group 1, n = 286 CVCs) and those prepared with conventional 10% povidone-iodine (group 2, n = 298 CVCs). RESULTS: Two hundred and thirty-nine patients received 584 CVCs for a total of 6,205 catheter-days. CVC-RBSI (3.22 per 1,000 catheter-days) was diagnosed in 20 cases. There were no significant differences in patient background factors between group 1 and 2, except for blood culture positivity (p = 0.0450). However, Kaplan-Meier analysis and the log rank test revealed no significant difference between group 1 and 2 in the time interval from insertion until development of CVC-RBSI. CONCLUSIONS: Use of 0.05% chlorhexidine is not inferior to conventional 10% povidone-iodine as a cutaneous disinfectant for the prevention of CVC-RBSI.

Is long-term tocolysis effective for threatened premature labour?

There is some controversy regarding the optimum duration of tocolysis. This retrospective multicentre study was performed to evaluate whether long-term (> 2 days) tocolysis is effective in treating threatened premature labour. A total of 1147 eligible patients were grouped according to whether or not tocolytics were given, and according to route of administration and whether or not ritodrine (the standard tocolytic regimen in Japan) or other tocolytic was given. They were then further stratified into three subgroups, using the Baumgarten and Gruber tocolysis index (TI), to assess the efficacy of tocolysis treatment according to the risk of premature labour. Prolongation of gestation was significantly longer in patients treated with tocolytics for > 2 days compared with the unmedicated, bed-rest group. In women receiving tocolysis, the mean duration of prolonged gestation was 2.2 times longer in the intermediate-risk TI group and 3.1 times longer in the high-risk TI group compared with the unmedicated group. In the patients who received tocolysis, IV ritodrine was used in 86% of cases and was considered safe and effective for prolonging gestation in cases of threatened premature labour.

Effect of maternal stress on fetal heart rate assessed by vibroacoustic stimulation.

This study was designed to determine whether maternal stress levels, state and trait anxiety levels, and stress hormones affect fetal heart rate (FHR) patterns after vibroacoustic stimulation (VAS) at 30 weeks of gestation. A total of 24 healthy pregnant women with a single fetus pregnancy were enrolled. Corticotropin releasing hormone (CRH) and adrenocorticotropic hormone in maternal plasma and cortisol, and chromogranin A in saliva were measured. The FHR patterns after VAS were divided into three types: type I, a long period of acceleration or one acceleration lasting > 1 min or at least two accelerations lasting > 15 s; type II, a biphasic response with acceleration followed by deceleration; and type III, no response or prolonged deceleration. In the high trait anxiety group, CRH levels were significantly higher than in the low trait anxiety group, and FHR patterns after VAS showed mostly a type II response pattern. These findings suggest that stress in pregnant women with high trait anxiety may influence FHR patterns after VAS.