Simultaneous gene transfer of bone morphogenetic protein (BMP)-2 and BMP-7 by in vivo electroporation induces rapid bone formation and BMP-4 expression

Background: Transcutaneous in vivo electroporation is expected to be an effective gene-transfer method for promoting bone regeneration using the BMP-2 plasmid vector. To promote enhanced osteoinduction using this method, we simultaneously transferred cDNAs for BMP-2 and BMP-7, as inserts in the non-viral vector pCAGGS. Methods: First, an in vitro study was carried out to confirm the expression of BMP-2 and BMP-7 following the double-gene transfer. Next, the individual BMP-2 and BMP-7 plasmids or both together were injected into rat calf muscles, and transcutaneous electroporation was applied 8 times at 100 V, 50 msec. Results: In the culture system, the simultaneous transfer of the BMP-2 and BMP-7 genes led to a much higher ALP activity in C2C12 cells than did the transfer of either gene alone. In vivo, ten days after the treatment, soft X-ray analysis showed that muscles that received both pCAGGS-BMP-2 and pCAGGS-BMP-7 had better-defined opacities than those receiving a single gene. Histological examination showed advanced ossification in calf muscles that received the double-gene transfer. BMP-4 mRNA was also expressed, and RT-PCR showed that its level increased for 3 days in a timedependent manner in the double-gene transfer group. Immunohistochemistry confirmed that BMP- 4-expressing cells resided in the matrix between muscle fibers. Conclusion: The simultaneous transfer of BMP-2 and BMP-7 genes using in vivo electroporation induces more rapid bone formation than the transfer of either gene alone, and the increased expression of endogenous BMP-4 suggests that the rapid ossification is related to the induction of BMP-4.</p

Loss of symbiont infectivity following thermal stress can be a factor limiting recovery from bleaching in cnidarians

Increases in seawater temperature can cause coral bleaching through loss of symbiotic algae (dinoflagellates of the family Symbiodiniaceae). Corals can recover from bleaching by recruiting algae into host cells from the residual symbiont population or from the external environment. However, the high coral mortality that often follows mass-bleaching events suggests that recovery is often limited in the wild. Here, we examine the effect of pre-exposure to heat stress on the capacity of symbiotic algae to infect cnidarian hosts using the Aiptasia (sea-anemone)-Symbiodiniaceae model system. We found that the symbiont strainBreviolumsp. CS-164 (ITS2 type B1), both free-living and in symbiosis, loses the capacity to infect the host following exposure to heat stress. This loss of infectivity is reversible, however, a longer exposure to heat stress increases the time taken for reversal. Under the same experimental conditions, the loss of infectivity was not observed in another strainBreviolum psygmophilumCCMP2459 (ITS2 type B2). Our results suggest that recovery from bleaching can be limited by the loss of symbiont infectivity following exposure to heat stress

Association between masticatory ability and oral functions

Mastication is the process of breaking ingested food with the teeth and mixing it with saliva to form a mass that is easy to swallow. However, few studies have reported on oral functions, such as occlusal force, tongue pressure, and mastication. The purpose of this study was to evaluate the association between masticatory function and oral functions, such as occlusal force and tongue pressure. In this study, there were 113 patients (41 men and 72 women; mean age, 68.4 ± 11.3 years) who visited dentists at the Hiroshima University Hospital, Hiroshima, Japan between April 2015 and November 2018. Masticatory function of the patients was evaluated using a masticatory ability test system. In addition, occlusal force was measured using a pressure-sensitive film and the maximum tongue pressure was measured with a tongue pressure measuring device according to a conventional method. The relationship of masticatory ability with occlusal force and tongue pressure was examined using multivariate analysis while considering patients? age, gender, and the number of remaining teeth. Masticatory ability was significantly related to occlusal force, maximum tongue pressure, age, body mass index, the number of remaining teeth, and occlusal contact area (p< 0.05). Multiple regression analysis identified that masticatory ability was significantly associated (p< 0.05) with occlusal force and maximum tongue pressure. Masticatory ability was significantly associated with occlusal force and maximum tongue pressure, indicating that the large muscle mass in the oral cavity is indispensable for improving masticatory function

Periodontal Tissue as a Biomaterial for Hard-Tissue Regeneration following bmp-2 Gene Transfer

The application of periodontal tissue in regenerative medicine has gained increasing interest since it has a high potential to induce hard-tissue regeneration, and is easy to handle and graft to other areas of the oral cavity or tissues. Additionally, bone morphogenetic protein-2 (BMP-2) has a high potential to induce the differentiation of mesenchymal stem cells into osteogenic cells. We previously developed a system for a gene transfer to the periodontal tissues in animal models. In this study, we aimed to reveal the potential and efficiency of periodontal tissue as a biomaterial for hard-tissue regeneration following a bmp-2 gene transfer. A non-viral expression vector carrying bmp-2 was injected into the palate of the periodontal tissues of Wistar rats, followed by electroporation. The periodontal tissues were analyzed through bone morphometric analyses, including mineral apposition rate (MAR) determination and collagen micro-arrangement, which is a bone quality parameter, before and after a gene transfer. The MAR was significantly higher 3–6 d after the gene transfer than that before the gene transfer. Collagen orientation was normally maintained even after the bmp-2 gene transfer, suggesting that the bmp-2 gene transfer has no adverse effects on bone quality. Our results suggest that periodontal tissue electroporated with bmp-2 could be a novel biomaterial candidate for hard-tissue regeneration therapy.Kawai M.Y., Ozasa R., Ishimoto T., et al. Periodontal Tissue as a Biomaterial for Hard-Tissue Regeneration following bmp-2 Gene Transfer. Materials, 15, 3, 993. https://doi.org/10.3390/ma15030993

Analysis of mineral apposition rates during alveolar bone regeneration over three weeks following transfer of BMP-2/7 gene via in vivo electroporation

Alveolar bone is not spontaneously regenerated following trauma or periodontitis. We previously proposed an animal model for new alveolar bone regeneration therapy based on the non-viral BMP-2/7 gene expression vector and in vivo electroporation, which induced the formation of new alveolar bone over the course of a week. Here, we analysed alveolar bone during a period of three weeks following gene transfer to periodontal tissue. Non-viral plasmid vector pCAGGS-BMP-2/7 or pCAGGS control was injected into palatal periodontal tissue of the first molar of the rat maxilla and immediately electroporated with 32 pulses of 50 V for 50 msec. Over the following three weeks, rats were double bone-stained by calcein and tetracycline every three days and mineral apposition rates (MAR) were measured. Double bone-staining revealed that MAR of alveolar bone was as similar level three days before BMP-2/7 gene transfer as three days after gene transfer. However, from 3 to 6 days, 6 to 9 days, 9 to 12 days, 12 to 15 days, 15 to 18 days, and 18 to 20 days after, MARs were significantly higher than prior to gene transfer. Our proposed gene therapy for alveolar bone regeneration combining non-viral BMP-2/7 gene expression vector and in vivo electroporation could increase alveolar bone regeneration potential in the targeted area for up to three weeks

IL-12 and IL-18 Induction and Subsequent NKT Activation Effects of the Japanese Botanical Medicine Juzentaihoto

In this study, we first measured some cytokine concentrations in the serum of patients treated with Juzentaihoto (JTT). Of the cytokines measured interleukin (IL) -18 was the most prominently up-regulated cytokine in the serum of patients under long term JTT administration. We next evaluated the effects of JTT in mice, focusing especially on natural killer T (NKT) cell induction. Mice fed JTT were compared to control group ones. After sacrifice, the liver was fixed, embedded and stained. Transmission electron microscope (TEM) observations were performed. Although the mice receiving the herbal medicine had same appearance, their livers were infiltrated with massive mononuclear cells, some of which were aggregated to form clusters. Immunohistochemical staining revealed that there was abundant cytokine expression of IL-12 and IL-18 in the liver of JTT treated mice. To clarify what the key molecules that induce immunological restoration with JTT might be, we next examined in vitro lymphocyte cultures. Mononuclear cells isolated and prepared from healthy volunteers were cultured with and without JTT. Within 24 hours, JTT induced the IL-12 and IL-18 production and later (72 hours) induction of interferon (IFN)-gamma. Oral administration of JTT may induce the expression of IL-12 in the early stage, and IL-18 in the chronic stage, followed by NKT induction. Their activation, following immunological restoration could contribute to anti-tumor effects

Management of Natural History collections: criteria and parameters of evaluation

[ES] Las colecciones de historia natural son una herramienta básica para la investigación científica y el estudio de la distribución en el pasado de muchas especies, así como de la propia historia de la ciencia. Además del uso científico de estas colecciones, destacan otros como el histórico, el divulgativo-pedagógico y el estético. De ahí la importancia que tiene una gestión eficaz de las mismas, la cual implica diversos aspectos, que van desde la conservación y su mantenimiento, su inventario, ordenación y procesamiento informático hasta las múltiples tareas relacionadas con su uso en consultas, visitas, préstamos científicos y participación en actividades de carácter divulgativo. En este artículo se examinan los criterios para evaluar la gestión de las colecciones de historia natural y se definen una serie de parámetros, útiles para medir el estado de una colección y su evolución en el tiempo, tanto en su crecimiento como en su uso, principalmente. Se ejemplifica todo ello en el grupo de los poliquetos, del que el Museo Nacional de Ciencias Naturales de Madrid (MNCN) cuenta con una estimable colección, y se comparan los resultados, según varios parámetros seleccionados, con los de otras colecciones de poliquetos de diversas instituciones de todo el mundo.[EN] Natural History collections are a basic and essential tool for scientific research, the study of the distribution in the past of many species of animals and plants and the History of Science. As well as the scientific aspect of these collections, stand outs other uses as the historic one, the educational and the aesthetic. All these are reasons that show the importance of an effective management of the Natural History collections as well as the several tasks related to it, as consults, visits, scientific loans and educational activities. In this article various criteria and useful parameters are provided for evaluating the curatorial state of a Natural History collection and its evolution, both in growth and use mainly. Finally, an example based on the Polychaeta, an estimable group in the Invertebrates Collection of the Museo Nacional de Ciencias Naturales of Madrid (MNCN), is provided. The results of several selected parameters are compared with other Polychaeta collections from several institutions around the world.Peer reviewe

Characteristic of GAFCHROMIC XR TYPE T dosimetry film

医療の高度化に伴い,近年では長時間のⅩ線透視を行うInterventional Radiology(IVR)手技が頻繁に行われ,副作用としての難治性放射線皮膚障害例の報告が増加している｡確定的影響である放射線皮膚障害はしきい値を超えると発症し,線量に依存して障害の程度が重篤となるため,患者被曝線量の測定が重要である｡しかし,IVRでは照射部位が多彩で,焦点-皮膚間距離が不安定なため,その測定方法は確立されていない｡本研究では,近年IVR等低エネルギー線量測定用フイルムとして開発されたCAFCHROMIC XR TYPE T について性能評価を行い,患者皮膚入射面の被曝線量測定への応用の可能性について検討した｡その結果,線量特性,線質特性,ネット値の安定性に良好な特性を示し,臨床に使用可能であったので報告する｡In recent years, interventional radiology (IVR) using which uses prolomged fluoroscopy has been performed frequently in clinical radiology. Also, reports of radiation skin injuries whose symptoms occur after IVR has been also increasing. These symptoms will become worse if the radiation induced skin injuries are caused by doses which are above the designated threshold, and the grade of injuries are dependent on dose. Therefore, it is important that patient skin dose is measured correctly. But when perfoming IVR, irradiation is complex pricedure, and there is a measurement error with an unfixed source-to-skin distance. So. in this paper, characteristics of GAFCHROMIC XR TYPE T (which are film for low energy X-rays) are performed. Then, they are decided whether the application to the skin surface incidence dose measurement whoud be posible. From these results usefull data can be obtainned; for exanmpe film characteristics, energy factors and stability of sensivility

Low incidence of limb-girdle muscular dystrophy type 2C revealed by a mutation study in Japanese patients clinically diagnosed with DMD

<p>Abstract</p> <p>Background</p> <p>Limb-girdle muscular dystrophy type 2C (LGMD2C) is an autosomal recessive muscle dystrophy that resembles Duchenne muscular dystrophy (DMD). Although DMD is known to affect one in every 3500 males regardless of race, a widespread founder mutation causing LGMD2C has been described in North Africa. However, the incidence of LGMD2C in Japanese has been unknown because the genetic background remains uncharacterized in many patients clinically diagnosed with DMD.</p> <p>Methods</p> <p>We enrolled 324 patients referred to the Kobe University Hospital with suspected DMD. Mutations in the dystrophin or the SGCG genes were analyzed using not only genomic DNA but also cDNA.</p> <p>Results</p> <p>In 322 of the 324 patients, responsible mutations in the dystrophin were successfully revealed, confirming DMD diagnosis. The remaining two patients had normal dystrophin expression but absence of γ-sarcoglycan in skeletal muscle. Mutation analysis of the SGCG gene revealed homozygous deletion of exon 6 in one patient, while the other had a novel single nucleotide insertion in exon 7 in one allele and deletion of exon 6 in the other allele. These mutations created a stop codon that led to a γ-sarcoglycan deficiency, and we therefore diagnosed these two patients as having LGMD2C. Thus, the relative incidence of LGMD2C among Japanese DMD-like patients can be calculated as 1 in 161 patients suspected to have DMD (2 of 324 patients = 0.6%). Taking into consideration the DMD incidence for the overall population (1/3,500 males), the incidence of LGMD2C can be estimated as 1 per 560,000 or 1.8 per million.</p> <p>Conclusions</p> <p>To the best of our knowledge, this is the first study to demonstrate a low incidence of LGMD2C in the Japanese population.</p