Dietary olive oil intake induces female-specific hepatic lipid accumulation without metabolic impairment in mice.

Olive oil is one of the most widely researched Mediterranean diet components in both experimental models and clinical studies. However, the relationship between dietary olive oil intake and liver function in a healthy state of the body remains unclear. Because men are at a greater risk of developing hepatic diseases than women, and because hepatic metabolism is regulated by sex hormones, we hypothesized that olive oil-induced changes in hepatic metabolism would differ by sex. To test our hypothesis, 12-week-old C57BL/6JJcl male and female mice were fed an olive oil diet for 4 weeks. Blood was collected and serum biochemical components were analyzed. Hepatic lipid accumulation was determined via histological analysis using Sudan III staining. Finally, transcript expression levels of hepatic metabolism-related genes were analyzed using quantitative polymerase chain reaction. We observed significant increased hepatic lipid droplet accumulation in olive oil-fed female mice. Serum biochemical and liver messenger RNA expression analyses revealed that the hepatic lipid accumulation was nonpathological and did not involve inflammation. Moreover, the expression of genes related to triacylglycerol and fatty acid synthesis (Dgat1, Dgat2, Agpat3, and Fasn) was significantly upregulated in the liver of olive oil-fed female mice compared with control female mice. Our study demonstrates female-specific hepatic lipid accumulation without liver impairment in a dietary olive oil-fed mouse model. These findings provide a deeper mechanistic understanding of sex-dependent hepatic lipid metabolism of dietary oils.

Streamlined human antibody generation and optimization by exploiting designed immunoglobulin loci in a B cell line.

Monoclonal antibodies (mAbs) are widely utilized as therapeutic drugs for various diseases, such as cancer, autoimmune diseases, and infectious diseases. Using the avian-derived B cell line DT40, we previously developed an antibody display technology, namely, the ADLib system, which rapidly generates antigen-specific mAbs. Here, we report the development of a human version of the ADLib system and showcase the streamlined generation and optimization of functional human mAbs. Tailored libraries were first constructed by replacing endogenous immunoglobulin genes with designed human counterparts. From these libraries, clones producing full-length human IgGs against distinct antigens can be isolated, as exemplified by the selection of antagonistic mAbs. Taking advantage of avian biology, effective affinity maturation was achieved in a straightforward manner by seamless diversification of the parental clones into secondary libraries followed by single-cell sorting, quickly affording mAbs with improved affinities and functionalities. Collectively, we demonstrate that the human ADLib system could serve as an integrative platform with unique diversity for rapid de novo generation and optimization of therapeutic or diagnostic antibody leads. Furthermore, our results suggest that libraries can be constructed by introducing exogenous genes into DT40 cells, indicating that the ADLib system has the potential to be applied for the rapid and effective directed evolution and optimization of proteins in various fields beyond biomedicine.

A nitrogen-containing bisphosphonate inhibits osteoblast attachment and impairs bone healing in bone-compatible scaffold.

Compromised osteoblast attachment on hydroxyapatite could be involved in the development of bone healing failure. We developed a bone-compatible scaffold that mimics bone structure with sub-micron hydroxyapatite (HA) surfaces, so that we could evaluate the effects of nitrogen-containing bisphosphonate (N-BP) on osteoblast behavior and bone healing. Human osteoblasts were seeded onto the bone-compatible scaffold with or without N-BP, and cell attachment and spreading behavior were evaluated 4 and 24 h after seeding. Then, mineralization was evaluated at 7 and 14 days. The osteoconductive activity of the scaffold was evaluated by implantation for 3 and 6 weeks into a rat cranial bone defect. The numbers of osteoblasts and their diameters were significantly less in N-BP-binding scaffolds than in untreated scaffolds at 4 and 24 h. Mineralization were also significantly less in the N-BP-binding scaffolds than in controls at 7 and 14 days. In vivo study revealed bone formation in N-BP-binding scaffolds was significantly less than in untreated scaffolds at 3 and 6 weeks. These results suggest that N-BP-binding to HA inhibited osteoblast attachment and spreading, thereby compromising bone healing process in the injured bone defect site.

New criteria of burst suppression on electroencephalogram in dogs anesthetized with sevoflurane.

Burst suppression on electroencephalogram (EEG) is defined as suppression periods longer than 0.5 s during which the amplitude does not exceed 5 µV in human. The aims of this study were; 1) an attempt of creating new criteria of burst suppression in dogs; and 2) a survey on accuracy of sub-parameter of Bispectral index (BIS). Using a BIS monitor, suppression ratio (SRBIS) and raw-EEG data were recorded at 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, and 5.0% end-tidal sevoflurane concentration (ETSEV) in 6 beagle dogs. The minimum ETSEV at which burst suppression was visually confirmed (ETSEVBS) was determined. By applying various duration and voltage threshold to criteria, suppression ratio was calculated (SR). Using the minimum balanced error rate (BER), new criteria consisting of the minimum duration of 0.35 s and the maximum threshold of 2.25 µV that provided SR > 0 above ETSEVBS was screened. SR was set by these criteria (SRBER) and by manual inspection (SRTRUE). The median detection rate of SRBER/SRTRUE was a statistically significant increase (p < .01) compared to that of SRBIS/SRTRUE (77% and 17% at 3.5% ETSEV, 89% and 19% at 4.0% ETSEV, and 86% and 84% at 5.0% ETSEV, respectively). In addition, between SRBER and SRTRUE evaluated by regression and Bland-Altman analyses, there was a strong correlation (r = 0.967, p < .001) and a moderate agreement (Limits of agreement: -7.14 ±â€¯13.95). The method using BER may help to establish new criteria of burst suppression to grasp the excessive deep level of anesthesia.

Multidisciplinary approach for treatment of a dentigerous cyst - marsupialization, orthodontic treatment, and implant placement: a case report.

BACKGROUND: Dentigerous cysts are common odontogenic cysts associated with unerupted teeth. We describe a previously unreported case of a multidisciplinary approach using surgical, orthodontic, and implant treatment to establish the occlusion for a patient with a maxillary dentigerous cyst. CASE PRESENTATION: An 18-year-old Japanese woman visited our hospital with a chief complaint of gingival swelling in her anterior maxillary region, midline diastema, and tooth crowding. Her main symptom was this gingival swelling. A panoramic radiograph revealed a radiolucent area, 30 mm in diameter, round in shape, and with well-demarcated margins including the maxillary canine. Computed tomography revealed a cystic cavity filled with homogeneous fluid of the same density as water, and a distolingually inclined canine. Our clinical diagnosis was maxillary dentigerous cyst with an unerupted distolingually inclined canine. The selected treatment was marsupialization of the dentigerous cyst, followed by orthodontic traction of the unerupted canine, and simultaneous orthodontic treatment of the midline diastema and tooth crowding. The orthodontic traction failed because the canine did not erupt completely, and the canine was extracted. The treatment plan was then changed to implant treatment after the tooth crowding and midline diastema had been improved. Because the alveolar ridge width was inadequate, the implant was placed after a two-stage implant treatment; therefore, a satisfactory occlusion could be achieved. Our patient did not experience any complications, and the cyst has not recurred. A radiograph taken 7 years after marsupialization of the dentigerous cyst revealed that the cystic cavity had been replaced by new bone. CONCLUSIONS: In general, orthodontic traction of an unerupted tooth after marsupialization should be the best option. However, if orthodontic traction fails, a multidisciplinary approach involving implant treatment may be necessary. We describe a case in which a multidisciplinary approach involving surgical, orthodontic, and implant treatment was used to establish a satisfactory occlusion for a patient with a dentigerous cyst.

Anti-Semaphorin 3A neutralization monoclonal antibody prevents sepsis development in lipopolysaccharide-treated mice.

Semaphorin 3A (Sema3A), originally identified as a potent growth cone collapsing factor in developing sensory neurons, is now recognized as a key player in immune, cardiovascular, bone metabolism and neurological systems. Here we established an anti-Sema3A monoclonal antibody that neutralizes the effects of Sema3A both in vitro and in vivo. The anti-Sema3A neutralization chick IgM antibodies were screened by combining an autonomously diversifying library selection system and an in vitro growth cone collapse assay. We further developed function-blocking chick-mouse chimeric and humanized anti-Sema3A antibodies. We found that our anti-Sema3A antibodies were effective for improving the survival rate in lipopolysaccharide-induced sepsis in mice. Our antibody is a potential therapeutic agent that may prevent the onset of or alleviate symptoms of human diseases associated with Sema3A.

A cellular stress response (CSR) that interacts with NADPH-P450 reductase (NPR) is a new regulator of hypoxic response.

NADPH-P450 reductase (NPR) was previously found to contribute to the hypoxic response of cells, but the mechanism was not clarified. In this study, we identified a cellular stress response (CSR) as a new factor interacting with NPR by a yeast two-hybrid system. Overexpression of CSR enhanced the induction of erythropoietin and hypoxia response element (HRE) activity under hypoxia in human hepatocarcinoma cell lines (Hep3B), while knockdown of CSR suppressed them. This new finding regarding the interaction of NPR with CSR provides insight into the function of NPR in hypoxic response.

Hydroxyapatite coating for titanium fibre mesh scaffold enhances osteoblast activity and bone tissue formation.

This study investigated the bone regeneration properties of titanium fibre mesh as a tissue engineering material. A thin hydroxyapatite (HA) coating on the titanium fibre web was created using the developed molecular precursor method without losing the complex interior structure. HA-coated titanium fibre mesh showed apatite crystal formation in vitro in a human osteoblast culture. Titanium fibre mesh discs with or without a thin HA coating were implanted into rat cranial bone defects, and the animals were killed at 2 and 4 weeks. The in vivo experience revealed that the amount of newly formed bone was significantly higher in the HA-coated titanium fibre mesh than in the non-coated titanium fibre mesh 2 weeks after implantation. These results suggest that thin HA coating enhances osteoblast activity and bone regeneration in the titanium fibre mesh scaffold. Thin HA-coating improved the ability of titanium fibre mesh to act as a bone regeneration scaffold.

Time-dependence and visualization of TiO2 and Pt particle biodistribution in mice.

Micro- and nano-sized materials have received much attention regarding their biocompatibility and toxicity. To understand the influence of such materials on animals, it is very important to determine their internal distribution behavior. In this study, the biodistributions of Pt and TiO2 micro- and nano-sized particles in mice were estimated and visualized by X-ray scanning analytical microscopy and inductively coupled plasma-atomic emission spectroscopy. We also determined the effect of particle size, difference between metal and oxide, and time dependence for the distributions, because the biodistribution depends upon both the chemical character of materials and the size of particles. The results of the present study indicated that the difference in chemical character had a greater effect than did particle size. We predict that X-ray scanning analytical microscopy will be a useful method for studying biodistribution of micro- and nano-sized particles, because this method requires no labeling or treatment of the target particles.

Cinacalcet suppresses calcification of the aorta and heart in uremic rats.

High serum parathyroid hormone levels are associated with vascular calcification. Cinacalcet is a calcimimetic agent that inhibits parathyroid hormone secretion and is used to treat patients with secondary hyperparathyroidism. Here we measured the effects of oral cinacalcet on calcification of the aorta and heart in rats with a remnant kidney (5/6 nephrectomy) model of uremia that were fed a high-phosphate diet containing lactose to accelerate the process of aortic calcification. Alizarin red staining showed that the smooth muscle in the aortic arch of rats with a remnant kidney was calcified. The tissue levels of calcium and phosphorus in the aorta and hearts of these rats were significantly increased compared to sham-operated rats. Expression of the osteoblastic lineage genes osteocalcin, osteopontin and runt-related gene 2 were also increased in the aorta of these rats. Cinacalcet suppressed these calcification-related changes by reducing serum parathyroid hormone, calcium, phosphorus, and the calcium-phosphorus product. Parathyroidectomy also suppressed calcification in this model. We suggest that cinacalcet inhibits calcification of the aorta and heart in uremic patients with secondary hyperparathyroidism by decreasing serum parathyroid hormone levels.

Down-regulation of the internal ribosome entry site (IRES)-mediated translation of the hepatitis C virus: critical role of binding of the stem-loop IIId domain of IRES and the viral core protein.

In a previous study, we observed that hepatitis C virus (HCV) core protein specifically inhibits translation initiated by an HCV internal ribosome entry site (IRES). To investigate the mechanism by which down-regulation of HCV translation occurs, a series of mutations were introduced into the IRES element, as well as the core protein, and their effect on IRES activity examined in this study. We found that expression of the core protein inhibits HCV translation possibly by binding to a stem-loop IIId domain, particularly a GGG triplet within the hairpin loop structure of the domain, within the IRES. Basic-residue clusters located at the N-terminus of the core protein have an inhibitory effect on HCV translation, and at least one of three known clusters is required for inhibition. We propose a model in which competitive binding of the core protein for the IRES and 40S ribosomal subunit regulates HCV translation.