Mesenchymal progenitor cells in adult human dental pulp and their ability to form bone when transplanted into immunocompromised mice

The technique of tissue engineering is developing for the restoration of lost tissues. This new technique requires cells that fabricate tissue. Mesenchymal stem cells in bone marrow have been used as the cell source for this technique; however, dental pulp cells have recently been shown to possess stem-cell-like properties. We earlier demonstrated that dental pulp cells proliferate and produce an extracellular matrix that subsequently becomes mineralized in vitro. We now report that such dental pulp cells (first to eighth passage) produced bone instead of dentin when those cells were implanted into subcutaneous sites in immunocompromised mice with HA/TCP powder as their carrier. This evidence shows that dental pulp cells are the common progenitors of odontoblasts and osteoblasts, or dental pulp cells are mesenchymal stem cells themselves. It is expected that dental pulp cells can be a useful candidate cell source for tissue engineering, and contain the potential of new therapeutic approaches for the restoration of damaged or diseased tissue.     

Distribution pattern and competition of insects breeding on Camellia japonica flower

The distribution pattern and competition of insects exploiting Camellia japonica flowers were studied in Tokyo, central Japan, to understand how their distributions are determined. Dasiops sp. of Lonchaeidae (Diptera) exploited flower buds and showed random distribution, whereas Drosophila unipectinata, D. oshimai and D. lutescens of Drosophilidae (Diptera) and Epuraea commutata of Nitidulidae (Coleoptera) exploited fully opened, late and fallen flowers and showed aggregated distribution. From the distribution pattern, it is assumed that Dasiops sp. has clutches of single egg whereas drosophilid and nitidulid species have clutches of more than one egg. In resource supplementary experiments, body size of drosophilid flies increased if resources were supplemented, although their survival is assumed to be unaffected. However, their body size did not decrease with increase of larval density in resource patches. It is therefore unclear whether resource competition occurs among drosophilid flies in Camellia flowers in nature. From the present and previous studies, it is assumed that aggregation (or production of clutches of more than one egg) is related to the use of fermenting or decayed resources; aggregation might increase larval survival and/or performance under the presence of molds or microorganisms.     

NRLMFβ: Beta-distribution-rescored neighborhood regularized logistic matrix factorization for improving the performance of drug–target interaction prediction

Techniques for predicting interactions between a drug and a target (protein) are useful for strategic drug repositioning. Neighborhood regularized logistic matrix factorization (NRLMF) is one of the state-of-the-art drug–target interaction prediction methods; it is based on a statistical model using the Bernoulli distribution. However, the prediction is not accurate when drug–target interaction pairs have less interaction information (e.g., the sum of the number of ligands for a target and the number of target proteins for a drug). This study aimed to address this issue by proposing NRLMF with beta distribution rescoring (NRLMFβ), which is an algorithm to improve the score of NRLMF. The score of NRLMFβ is equivalent to the value of the original NRLMF score when the concentration of the beta distribution becomes infinity. The beta distribution is known as a conjugative prior distribution of the Bernoulli distribution and can reflect the amount of interaction information to its shape based on Bayesian inference. Therefore, in NRLMFβ, the beta distribution was used for rescoring the NRLMF score. In the evaluation experiment, we measured the average values of area under the receiver operating characteristics and area under precision versus recall and the 95% confidence intervals. The performance of NRLMFβ was found to be better than that of NRLMF in the four types of benchmark datasets. Thus, we concluded that NRLMFβ improved the prediction accuracy of NRLMF. The source code is available at https://github.com/akiyamalab/NRLMFb.     

Preliminary evaluation of greenhouses employing positive-pressure forced ventilation to prevent invasion by insect pests

We conducted a preliminary comparison of greenhouses using positive-pressure forced ventilation (PFV) systems and natural ventilation (NV) systems, and assessed the effectiveness of both systems for preventing the invasion of greenhouses used to cultivate tomatoes by insect pests. In Trial 1 (August–December 2006), greenhouses using a PFV system and an insect-proof screen (mesh size 1.0 mm) had fewer sweetpotato whiteflies and Bemisia tabaci (Gennadius), and more onion thrips, Thrips tabaci (Lindeman), than greenhouses that employed an NV system fitted with the same screen. Tomato leafminers, Liriomyza sativae Blanchard, were not observed in the greenhouse using the PFV system, but some were observed in the greenhouse using the NV system. In Trial 2 (August–December 2007), the greenhouse using the PFV system combined with an insect-proof screen (mesh size 0.4 mm) had higher whitefly densities after late October compared to the greenhouse using the NV system and the same screen. However, there were more thrips in the greenhouse using the PFV system compared to the greenhouse using the NV system. In both trials, Tomato yellow leaf curl virus, which is transmitted by B. tabaci, was recorded in the greenhouse using the NV system but not in the greenhouse using the PFV system. The results showed that the PFV system was effective for preventing invasion by leafminers and partially effective for preventing invasion by whiteflies, but not effective for preventing invasion by thrips.     

Netrin-4 derived from murine vascular endothelial cells inhibits osteoclast differentiation in vitro and prevents bone loss in vivo

Bone is a highly vascularized organ, thus angiogenesis is a vital process during bone remodeling. However, the role of vascular systems in bone remodeling is not well recognized. Here we show that netrin-4 inhibits osteoclast differentiation in vitro and in vivo. Co-cultures of bone marrow macrophages with vascular endothelial cells markedly inhibited osteoclast differentiation. Adding a neutralizing antibody, or RNA interference against netrin-4, restored in vitro osteoclast differentiation. Administration of netrin-4 prevented bone loss in an osteoporosis mouse model by decreasing the osteoclast number. We propose that vascular endothelial cells interact with bone in suppressing bone through netrin-4.     

Congenital malformations in Japanese macaques (Macaca fuscata) at Takasakiyama

From the late 1960s to the early 1970s, many congenitally malformed infants were born into provisioned Japanese macaque troops. Although the exact cause of this problem was not determined, the occurrence of malformations decreased thereafter. We examined possible factors such as total population size, number of adult females, birth rate, and volume of provisioned food. Agrichemicals attached to provisioned food are suspected as the main cause, as other factors were found to have no influence. Many more malformations were seen in males compared with females, in feet compared with hands, and in the fourth compared with other digits. We confirmed that the frequency of congenital malformation was high during the 1960s through to the mid-1970s when increased levels of provisioned food were given and that the incidence of congenital malformations was also elevated among wild macaques during this time.     

Sperm guidance to the egg finds calcium at the helm

Sperm respond to multiple cues during guidance to the egg including chemical attractants, temperature, and fluid flow. Of these, sperm chemotaxis has been studied most extensively—over 100 years—but only recently has it started to be understood at the molecular level. The long gestation in this understanding has largely been due to technical limitations that include the detection of calcium signal dynamics in a relatively small structure—the flagellum, measurement of actual chemoattractant gradients, the fact that only subpopulations of sperm respond at any given time, and the diversity in swimming behaviors that sperm exhibit from different species. Today, measurements of flagellar calcium signals on a fast time scale, discovery of the ion channels and organelles that may regulate these signals, and better understanding and quantitation of sperm swimming behaviors involved have given more certainty to our understanding of sperm directional swimming and its control by characteristic, calcium-directed asymmetric flagellar bends. Future research will need to apply these technical advances to other forms of sperm guidance such as thermotaxis and rheotaxis as well as gaining an understanding of how the flagellar apparatus is controlled by calcium.     

Locating the uracil-5-yl radical formed upon photoirradiation of 5-bromouracil-substituted DNA

In a previous study, we found that 2-deoxyribonolactone is effectively generated in the specific 5-bromouracil (^BrU)-substituted sequence 5′-(G/C)[A]_{n = 1,2}^BrU^BrU-3′ and proposed that a formed uracil-5-yl radical mainly abstracts the C1′ hydrogen from the 5′-side of ^BrU^BrU under 302-nm irradiation condition. In the present work, we performed photoirradiation of ^BrU-substituted DNA in the presence of a hydrogen donor, tetrahydrofuran, to quench the uracil-5-yl radical to uracil and then subjected the sample to uracil DNA glycosylase digestion. Slab gel sequence analysis indicated that uracil residues were formed at the hot-spot sequence of 5′-(G/C)[A]_{n = 1,2}^BrU^BrU-3′ in 302-nm irradiation of ^BrU-substituted DNA. Furthermore, we found that the uracil residue was also formed at the reverse sequence 5′-^BrU^BrU[A]_{n = 1,2}(G/C)-3′, which suggests that both 5′-(G/C)[A]_{n = 1,2}^BrU^BrU-3′ and 5′-^BrU^BrU[A]_{n = 1,2}(G/C)-3′ are hot-spot sequences for the formation of the uracil-5-yl radical.     

Simple generation of albino C57BL/6J mice with G291T mutation in the tyrosinase gene by the CRISPR/Cas9 system

Single nucleotide mutations (SNMs) are associated with a variety of human diseases. The CRISPR/Cas9 genome-editing system is expected to be useful as a genetic modification method for production of SNM-induced mice. To investigate whether SNM-induced mice can be generated by zygote microinjection of CRISPR/Cas9 vector and single-stranded DNA (ssDNA) donor, we attempted to produce albino C57BL/6J mice carrying the Tyr gene SNM (G291T) from pigmented C57BL/6J zygotes. We first designed and constructed a CRISPR/Cas9 expression vector for the Tyr gene (px330-Tyr-M). DNA cleavage activity of px330-Tyr-M at the target site of the Tyr gene was confirmed by the EGxxFP system. We also designed an ssDNA donor for homology-directed repair (HDR)-mediated gene modification. The px330-Tyr-M vector and ssDNA donor were co-microinjected into the pronuclei of 224 one-cell-stage embryos derived from C57BL/6J mice. We obtained 60 neonates, 28 of which showed the ocular albinism and absence of coat pigmentation. Genomic sequencing analysis of the albino mice revealed that the target of SNM, G291T in the Tyr gene, occurred in 11 mice and one founder was homozygously mutated. The remaining albino founders without Tyr G291T mutation also possessed biallelic deletion and insertion mutants adjacent to the target site in the Tyr locus. Simple production of albino C57BL/6J mice was provided by C57BL/6J zygote microinjection with px330-Tyr-M DNA vector and mutant ssDNA (G291T in Tyr) donor. A combination of CRISPR/Cas9 vector and optional mutant ssDNA could be expected to efficiently produce novel SNM-induced mouse models for investigating human diseases.