Isolation and artificial production of atypical porcine pestivirus, using the swine-kidney-derived cell line SK-L.

Congenital tremor (CT) in piglets was first reported in 1922, and although the causative pathogen was unknown for many years, atypical porcine pestivirus (APPV) was recently shown to be the cause. APPV is difficult to isolate, and there have been few reports of APPV isolated from field materials. Here, we successfully isolated infectious particles from a tonsillar emulsion from a CT-affected piglet using the established swine-kidney-derived cell line SK-L. In addition, we produced APPV artificially using these cells. Thus, SK-L cells are useful for both isolation and artificial production of APPV.

Division of loading time in reloading the disused atrophic soleus muscle induces proximal muscle injury.

[Purpose] This study aimed to compare the effects of loading time division in reloading atrophied muscles in different muscle long-axis regions. [Materials and Methods] We divided 8-week-old male Wistar rats into control (CON), 14-day hindlimb suspension (HS), 7-day hindlimb suspension followed by 60-min reloading for 7 consecutive days (WO), and 7-day hindlimb suspension followed by 60-min reloading on two separate occasions for 7 days (WT) groups. After the experimental period, muscle fibre cross-sectional area and necrotic fibre/central nuclei fibre ratio were measured in the soleus muscle's proximal, middle, and distal regions. [Results] The necrotic fibre/central nuclei fibre ratio was higher in the WT group than in the other groups in the proximal region. Proximal muscle fibre cross-sectional area was higher in the CON group than in the other groups. In the middle region, only HS group had muscle fibre cross-sectional area lower than the CON group. Similarly, muscle fibre cross-sectional area of the HS group was lower than the CON and WT groups in the distal region. [Conclusion] When reloading atrophied muscles, dividing the loading time can inhibit atrophy in the distal region but induce muscle injury in the proximal region.

Molecular cloning and characterization of type III polyketide synthase from Plumbago zeylanica.

Two cDNAs encoding type Ш polyketide synthase (PKS1) and chalcone synthase (CHS, PKS2), were cloned from fresh leaves of Plumbago zeylanica L. (P. zeylanica). Their heterologous expression revealed that PKS1 catalyzed the formation of five α-pyrones from three to six acetate units by accepting acetyl-CoA and malonyl-CoA. In contrast, PKS2 catalyzed the formation of naringenin and bisnoryangonin by accepting p-coumaroyl-CoA and malonyl-CoA. Naringenin is thought to be involved in the biosynthesis of various bioactive flavonoids. PKS2 can be used to molecular breeding to enhance the production of these useful secondary metabolites via its overexpression.[Formula: see text].

YAP activation in Müller cells protects against NMDA-induced retinal ganglion cell injury by regulating Bcl-xL expression.

Retinal neurodegeneration, characterized by retinal ganglion cell (RGC) death, is a leading cause of vision impairment and loss in blind diseases, such as glaucoma. Müller cells play crucial roles in maintaining retinal homeostasis. Thus, dysfunction of Müller cells has been implicated as one of the causes of retinal diseases. Yes-associated protein 1 (YAP), a nuclear effector of the Hippo pathway, regulates mammalian cell survival. In this study, we investigated the role of YAP in Müller cells during N-methyl-D-aspartic acid (NMDA)-induced excitotoxic RGC injury in rats. We found that YAP expression increased and was activated in Müller cells after NMDA-induced RGC injury. This YAP response was partly due to an increase in Yap mRNA levels, although it may be independent of the Hippo pathway and ß-TrCP-mediated YAP degradation. Morphological analysis revealed that verteporfin, a selective YAP inhibitor, exacerbated NMDA-induced RGC degeneration, suggesting that YAP activation in Müller cells contributes to RGC survival in NMDA-treated retinas. Studies in the rat Müller cell line (rMC-1) demonstrated that overexpression of YAP increased the levels of Bcl-xL, while verteporfin decreased the levels of Bcl-xL and cell viability and increased the levels of cytochrome c released from mitochondria and cleaved caspase-3. Finally, we found that Bcl-xL expression increased slightly in NMDA-treated retinas, whereas intravitreal injection of verteporfin suppressed this increase. Our findings suggest that activated YAP in Müller cells protects against NMDA-induced RGC injury by upregulating Bcl-xL expression.

Anterior cruciate ligament reconstruction with ultrasound-guided femoral nerve block does not adversely affect knee extensor strength beyond that seen with intravenous patient-controlled analgesia at 3 and 6 months postoperatively.

BACKGROUND: This study aimed to investigate the effect of anterior cruciate ligament (ACL) reconstruction with an ultrasound-guided femoral nerve block (FNB) on knee extensor strength weakness 3 and 6 months, and graft rupture in the 1 year following ACL reconstruction. METHODS: One hundred and seven patients who underwent ACL reconstruction were included in this retrospective study. The patients were divided into two groups stratified by the method of postoperative pain management. The FNB group included 66 patients, and there were 41 patients in the intravenous patient-controlled analgesia (iv-PCA) group. The isokinetic peak torque of knee flexor and extensor was measured preoperative, 3 and 6 months after ACL reconstruction. Muscle strength measurements were performed using the BIODEX dynamometer at a velocity of 60°/s and 180°/s. Peak torque of knee extensor and flexor strength, estimated pre-injury capacity (EPIC), body weight ratio (BW), and graft rupture incidence were compared between the two groups. RESULTS: There were no statistically significant differences in the knee extensor and flexor strength for all items at 3 and 6 months after ACL reconstruction. There was also not a statistically significant difference in the graft rupture incidence between the two groups: FNB group was two patients, 3.0% vs. iv-PCA group was one patient, 2.4% (p = 0.86). CONCLUSION: ACL reconstruction with ultrasound-guided FNB does not affect knee extensor strength at 6 months, nor graft rupture at 1 year postoperatively.

Magnetic particles-based enzyme immunoassay for rapid determination of secoiridoid glycoside, amarogentin.

Amarogentin (AG) is one of the bitter secoiridoid glycosides, which exerts various pharmacological activities as a bitter stomachic. Recently, there is an increasing demand for AG-containing plants in Japan due to their use as folk medicines and food additives; hence, it is crucial to develop analytical techniques that are specific for AG. In this study, a new magnetic particles-based enzyme immunoassay (MPs-EIA) using a specific monoclonal antibody against AG (MAb 1E9) for the rapid determination of AG in plants of the family Gentianaceae was described. AG directly immobilized onto magnetic particles (MPs) was used as a competitor for free AG against MAb 1E9, thereby increasing the surface area of the solid phase and decreasing the immunoreaction time. In addition, the blocking step required in case of the conventional enzyme-linked immunosorbent assay could be avoided in the proposed MPs-EIA, which enables an even more rapid performance for the immunoassay. In the developed MPs-EIA, AG exhibited linearity in the range of 15.6-500 ng mL-1, with a limit of detection of 8.58 ng mL-1. Validation analysis revealed that MPs-EIA is a sufficiently sensitive and rapid for the quantitative analysis of AG in plant samples. To the best of our knowledge, this is the first MPs-EIA that has been applied to plant samples.