Calcium Chloride Treatment Enhances Antigen Production in Foot-and-Mouth Disease Vaccines for Serotypes SAT1 and SAT3.

Foot-and-mouth disease (FMD) is a highly contagious viral infection causing acute and severe vesicular lesions in cattle and pigs, which has prompted global vaccination policies. This study presents a technique for enhancing antigen yield in SAT1 BOT and SAT3 ZIM by treatment with calcium chloride (CaCl2). We tested changes in cell viability in BHK-21 suspension cells treated with varying concentrations of CaCl2. The optimal CaCl2 concentration was determined based on antigen yield. The timing of CaCl2 supplementation relative to FMD virus inoculation was tested. Finally, the optimal medium for antigen production was identified. We observed a concentration-dependent decrease in BHK-21 cell viability at >7.5 mM CaCl2. A CaCl2 concentration of 3 mM yielded the most antigens. CaCl2 supplementation relative to FMD virus infection was optimal 2 h before or with viral inoculation. CD-BHK 21 medium supplemented with CaCl2 was the most productive medium. Specifically, SAT1 BOT and SAT3 ZIM showed improved antigen production in CD-BHK 21 medium with 3 mM CaCl2, while Provero-1 and Cellvento BHK-200 media showed no significant enhancement. Overall, CaCl2 supplementation enhanced FMD antigen productivity. This study provides a useful framework for enhancing antigen production efficiently in the FMD vaccine industry.

Types of Polyunsaturated Fatty Acids in Epididymal Adipose Tissue Are Distinguishable Using Raman Spectroscopy and Multivariate Analysis.

Biological polyunsaturated fatty acids (PUFAs) are important precursors of secondary messengers that modulate inflammatory responses, cellular growth, and cholesterol metabolism. The optimal n-6/n-3 ratio is extremely important for maintaining normal homeostasis because n-3 and n-6 PUFAs are competitively metabolized. To date, a widely accepted analytical method to determine the biological n-6/n-3 ratio is gas chromatography-mass spectrometry (GC-MS) on dried whole blood samples. However, this technique has several drawbacks, including the intrusive nature of collecting blood samples, high expenses involved, and length of time required to use the GC/MS instrument. To overcome these limitations, we introduced Raman spectroscopy (RS) to distinguish PUFAs present in the epididymal adipose tissue (EAT) isolated from experimental rats that were fed three different high-fat diets (HFDs) with multivariate analysis, including principal component analysis (PCA) and linear discriminant analysis (LDA). The diets comprised HFD, HFD + perilla oil (HFD + PO [n-3 rich oil]), and HFD + corn oil (HFD + CO [n-6 rich oil]). This method allows for quantitative, label-free, noninvasive, and rapid monitoring of biochemical changes in the EAT with high sensitivity. In RS, the Raman bands of the EAT from three different diet groups (HFD, HFD + PO, and HFD + CO) detected and distinguished peaks at 1079 (C-C stretching vibration), 1300 (CH2 deformation), 1439 (CH2 deformation), 1654 (amide I), 1746 (C = O stretching vibration), and 2879 cm-1 (-C-H stretching vibration). The PCA-LDA analysis results showed that PUFAs in the EAT of animals receiving the three different dietary interventions can be determined according to the three groups (HFD, HFD + PO, and HFD + CO). In conclusion, we investigated the possibility of determining PUFA profiles in specimens using RS.

Evaluation of the color stability of 3D printed resin according to the oxygen inhibition effect and temperature difference in the post-polymerization process.

The aim of this study was to determine the color stability of 3D printed resin according to the post-curing conditions (polymerization conditions and temperature). Specimens were post-polymerized under different conditions of oxygen inhibition, such as on glycerin immersion (GLY), medium-low vacuum environment (VA), and oxygen contact (CON, the control group), and temperature (35 °C, 60 °C, and 80 °C). The degree of conversion (DC), water sorption (Wsp) and solubility (Wsl), surface roughness (Ra) were measured. Additionally, surface free energy (SFE), pH values of colorants were measured. Grape juice (grape), coffee, and curry were used as the colorants, and distilled water (DW) was used as a control. And the color value was measured before and after immersion using a spectrophotometer. Then, Calculated the color change. For statistical methods, The Shapiro-Wilk test performed to check for normality revealed that the data presented a normal distribution (p>0.05). ΔE values were analyzed using three-way ANOVA. DC, Wsp, Wsl, SFE, and Ra were analyzed using two-way ANOVA. To confirm the linear correlation, Pearson's correlation coefficient was determined. The threshold for significance (p) was set at 0.05 (95% confidence interval) for all tests. DC was the highest at 80 °C in the GLY group (95.08 ± 4.88%). And Wsl decreased with increasing temperature, and was lowest at 80 °C in the GLY group (0.46 ± 0.30 um/mm3). After the colorants were immersed for 30 days, as the temperature increased, ΔE decreased in the GLY group but not in the VA and CON groups, and was the lowest at 80 °C in the GLY group: (DW, 0.95 ± 0.45 [mean± SD]; grape, 6.45± 0.69; coffee, 4.50± 0.56; curry, 9.37± 1.40). There was also a significant inverse relation between DC and ΔE. A significant inverse relation was found between Wsl and DC, and a significant positive correlation was found between Wsl and ΔE. Wsp, SFE, and Ra did not affect color stability. In the post-polymerization process, increasing the temperature and GLY were effective in reducing ΔE, which was lowest at 80 °C in the GLY group. It was also observed that a complex mechanism between the DC, Wsl of 3D printed resin affects ΔE of the resin.

Neuroprotective effect of Geijigadaehwang-tang against trimethyltin-induced hippocampal neurodegeneration: An in vitro and in vivo study.

ETHNOPHARMACOLOGICAL RELEVANCE: Patients with dementia are diagnosed with deficiency patterns and interior patterns in traditional Chinese medicine due to decreased physical strength, mental atrophy including cognitive function, and decreased motor function in the gastrointestinal tract. Since "greater yin symptom" in Shanghanlun has been interpreted as interior, deficiency, and cold pattern in traditional Chinese medicine, it is necessary to determine whether Geijigadaehwang-tang (GDT) has therapeutic effects on neurodegenerative diseases and the underlying mechanism if it has such effects. AIMS OF THE STUDY: Trimethyltin (TMT), a neurotoxic organotin compound, has been used to induce several neurodegenerative diseases, including epilepsy and Alzheimer's disease. This study aimed to evaluate the therapeutic efficacy of GDT for TMT-induced hippocampal neurodegeneration and seizures and to determine the mechanisms involved at the molecular level. MATERIALS AND METHODS: The main components of GDT were analyzed using ultra-performance liquid chromatography. TMT was used to induce neurotoxicity in microglial BV-2 cells and C57BL6 mice. GDT was administered at various doses to determine its neuroprotective and seizure inhibition effects. The inhibitory effects of GDT on TMT-induced apoptosis, inflammatory pathways, and oxidative stress pathways were determined in the mouse hippocampal tissues. RESULTS: GDT contained emodin, chrysophanol, albiflorin, paeoniflorin, 6-gingerol, and liquiritin apioside. In microglial BV-2 cells treated with TMT, GDT showed dose-dependent neuroprotective effects. Oral administration of GDT five times for 2.5 days before and after TMT injection inhibited seizures at doses of 180 and 540 mg/kg and inhibited neuronal death in the hippocampus. In hippocampal tissues extracted from mice, GDT inhibited the protein expression of ionized calcium binding adaptor molecule 1, glial fibrillary acidic protein, nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing protein 3, and phosphorylated nuclear factor (NF)-κB/total-NFκB ratio. Additionally, GDT inhibited the messenger RNA levels of tumor necrosis factor-α, inducible nitric oxide synthase, apoptosis-associated speck-like protein containing a caspase recruitment domain, caspase-1, interleukin-1ß, nuclear factor erythroid-2-related factor 2, and heme oxygenase-1. CONCLUSION: This study's results imply that GDT might have neuroprotective potential in neurodegenerative diseases through neuronal death inhibition and anti-inflammatory and antioxidant mechanisms.

Natural Product Ginsenoside 20(S)-25-Methoxyl-Dammarane-3ß, 12ß, 20-Triol in Cancer Treatment: A Review of the Pharmacological Mechanisms and Pharmacokinetics.

Panax ginseng has been used as an herbal medicine for thousands of years. Most of its pharmacological effects are attributed to its constituent ginsenosides, including 20(S)-25-methoxyl-dammarane-3ß, 12ß, 20-triol (20(S)-25-OCH3-PPD), which is one of the protopanaxadiol type ginsenosides. It has been found to exhibit anticancer effects by interacting with multiple pharmacological pathways, such as the Wnt/ß-catenin, MDM2, ERK/MAPK, and STAT3 signaling pathways. However, its therapeutic potential could be limited by its low bioavailability mainly due to its low aqueous solubility. Thus, several studies have been conducted on its pharmacokinetics and its delivery systems, so as to increase its oral bioavailability. In this review, comprehensive information on its varying pharmacological pathways in cancer, as well as its pharmacokinetic behavior and pharmaceutical strategies, is provided. This information would be useful in the understanding of its diverse mechanisms and pharmacokinetics as an anticancer drug, leading to the design of superior 20(S)-25-OCH3-PPD-containing formulations that maximize its therapeutic potential.

LRR domain of NLRX1 protein delivery by dNP2 inhibits T cell functions and alleviates autoimmune encephalomyelitis.

Multiple sclerosis (MS) is a demyelinating inflammatory disease of the central nervous system (CNS), which is a chronic progressive disease and is caused by uncontrolled activation of myelin antigen specific T cells. It has high unmet medical needs due to the difficulty of efficient drug delivery into the CNS to control tissue inflammation. In this study, we demonstrate that a fusion protein of NOD-like receptor family member X1 (NLRX1) and blood brain barrier (BBB)-permeable peptide, dNP2 ameliorates experimental autoimmune encephalomyelitis (EAE). Methods: We purified recombinant LRR or NBD regions of NLRX1 protein conjugated with dNP2. To examine intracellular delivery efficiency of the recombinant protein, we incubated the proteins with Jurkat T cells or murine splenic T cells and their delivery efficiency was analyzed by flow cytometry. To investigate the therapeutic efficacy in an EAE model, we injected the recombinant protein into mice with 3 different treatment schemes e.g., prevention, semi-therapeutic, and therapeutic. To analyze their functional roles in T cells, we treated MACS-sorted naïve CD4 T cells with the proteins during their activation and differentiation into Th1, Th17, and Treg cells. Results: dNP2-LRR protein treatment showed significantly higher delivery efficiency than TAT-LRR or LRR alone in Jurkat T cells and mouse splenic T cells. In all three treatment schemes of EAE experiments, dNP2-LRR administration showed ameliorated tissue inflammation and disease severity with reduced number of infiltrating T cells producing inflammatory cytokines such as IFNγ. In addition, dNP2-LRR inhibited T cell activation, cytokine production, and Th1 differentiation. Conclusion: These results suggest that dNP2-LRR is a novel agent, which regulates effector T cell functions and could be a promising molecule for the treatment of CNS autoimmune diseases such as multiple sclerosis.

Therapeutic Effects of Licorice and Dried Ginger Decoction on Activity-Based Anorexia in BALB/c AnNCrl Mice.

Licorice and dried ginger decoction (Gancao-ganjiang-tang, LGD) is used for nausea and anorexia, accompanied by excessive sweating in Traditional Chinese Medicine. Herein, we investigated the therapeutic effects of LGD using the activity-based anorexia (ABA) in a mouse model. Six-week-old female BALB/c AnNCrl mice were orally administered LGD, water, licorice decoction, dried ginger decoction, or chronic olanzapine, and their survival, body weight, food intake, and wheel activity were compared in ABA. Additionally, dopamine concentration in brain tissues was evaluated. LGD significantly reduced the number of ABA mice reaching the drop-out criterion of fatal body weight loss. However, LGD showed no significant effects on food intake and wheel activity. We found that in the LGD group the rise of the light phase activity rate inhibited body weight loss. Licorice or dried ginger alone did not improve survival rates, they only showed longer survival periods than chronic olanzapine when combined. In addition, LGD increased the dopamine concentration in the brain. The results from the present study showed that LGD improves the survival of ABA mice and its mechanism of action might be related to the alteration of dopamine concentration in the brain.

Novel anti-biofouling bioactive calcium silicate-based cement containing 2-methacryloyloxyethyl phosphorylcholine.

Calcium silicate-based cements (CSCs) are commonly used for endodontic procedures; however, their antibacterial effects are limited. The objective of this study was to develop a 2-methacryloyloxyethyl phosphorylcholine (MPC)-incorporated CSC with improved antibacterial properties, while maintaining the original advantageous features of CSC. MPC was incorporated into a commercial CSC (Endocem MTA) at 0 wt% (control), 1.5%, 3.0 wt%, 5.0 wt%, 7.5 wt%, and 10 wt%. The setting time, compressive strength, water sorption, and glycerol contact angle were measured. Protein absorption was measured and bacterial adhesion on the surface was evaluated using Enterococcus faecalis. The bactericidal effect was examined by the disc diffusion test. Mineralization ability was assessed based on calcium ion deposition, as assessed by alizarin red staining, after immersion into Hank's balanced salt solution for 7 days. High concentrations of MPC in CSC (7.5 wt% and 10 wt%) increased the setting time, reduced compressive strength, and reduced wettability. MPC (3 wt%) had greater protein repellent and anti-biofouling effects than those of control and test materials (P < 0.001). However, no bactericidal effect was observed for any control or test materials. There was greater calcium ion deposition on the surface of MPC-supplemented CSC than on the control (P < 0.001). The addition of 3 wt% MPC polymer to CSC confers protein-repellent properties and reduced bacterial attachment, with the potential for improved mineralization.

Zinc Promotes Osteoblast Differentiation in Human Mesenchymal Stem Cells Via Activation of the cAMP-PKA-CREB Signaling Pathway.

The crucial trace element zinc stimulates osteogenesis in vitro and in vivo. However, the pathways mediating these effects remain poorly understood. This study aimed to investigate the effects of zinc on osteoblast differentiation in human bone marrow-derived mesenchymal stem cells (hBMSCs) and to identify the molecular mechanisms of these effects. In hBMSCs, zinc exposure resulted in a dose-dependent increase in osteogenesis and increased mRNA and protein levels of the master transcriptional factor RUNX2. Analyzing the upstream signaling pathways of RUNX2, we found that protein kinase A (PKA) signaling inhibition blocked zinc-induced osteogenic effects. Zinc exposure increased transcriptional activity and protein levels of phospho-CREB and enhanced translocation of phospho-CREB into the nucleus. These effects were reversed by H-89, a potent inhibitor of PKA. Moreover, zinc exposure led to dose-dependent increases in levels of intracellular cyclic adenosine monophosphate (cAMP). These findings indicate that zinc activates the PKA signaling pathway by triggering an increase in intracellular cAMP, leading to enhanced osteogenic differentiation in hBMSCs. Our results suggest that zinc exerts osteogenic effects in hBMSCs by activation of RUNX2 via the cAMP-PKA-CREB signaling pathway. Zinc supplementation may offer a promise as a potential pharmaceutical therapy for osteoporosis and other bone loss conditions.

Nano-electromechanical Switch Based on a Physical Unclonable Function for Highly Robust and Stable Performance in Harsh Environments.

A physical unclonable function (PUF) device using a nano-electromechanical (NEM) switch was demonstrated. The most important feature of the NEM-switch-based PUF is its use of stiction. Stiction is one of the chronic problems associated with micro- and nano-electromechanical system (MEMS/NEMS) devices; however, here, it was utilized to intentionally implement a PUF for hardware-based security. The stiction is caused by capillary and van der Waals forces, producing strong adhesion, which can be utilized to design a highly robust and stable PUF. The probability that stiction will occur on either of two gates in the NEM switch is the same, and consequently, the occurrence of the stiction is random and unique, which is critical to its PUF performance. This uniqueness was evaluated by measuring the interchip Hamming distance (interchip HD), which characterizes how different responses are made when the same challenge is applied. Uniformity was also evaluated by the proportion of "1" or "0" in the response bit-string. The reliability of the proposed PUF device was assessed by stress tests under harsh environments such as high temperature, high dose radiation, and microwaves.

Therapeutic Effect of Chung-Pae, an Experimental Herbal Formula, on Acute Lung Inflammation Is Associated with Suppression of NF- κ B and Activation of Nrf2.

Acute lung injury (ALI) is an inflammatory disease with high mortality, but therapeutics against it is unavailable. Recently, we elaborated a formula, named Chung-pae (CP), that comprises four ethnic herbs commonly prescribed against various respiratory diseases in Asian traditional medicine. CP is being administered in aerosol to relieve various respiratory symptoms of patients in our clinic. Here, we sought to examine whether CP has a therapeutic effect on ALI and to uncover the mechanism behind it. Reporter assays show that CP suppressed the transcriptional activity of proinflammatory NF- κ B and activated that of anti-inflammatory Nrf2. Similarly, CP suppressed the expression of NF- κ B dependent, proinflammatory cytokines and induced that of Nrf2 dependent genes in RAW 264.7. An aerosol intratracheal administration of CP effectively reduced neutrophilic infiltration and the expression of pro-inflammatory cytokines, hallmarks of ALI, in the lungs of mice that received a prior intraperitoneal injection of lipopolysaccharide. The intratracheal CP administration concomitantly enhanced the expression of Nrf2 dependent genes in the lung. Therefore, our results evidenced a therapeutic effect of CP on ALI, in which differential regulation of the two key inflammatory factors, NF- κ B and Nrf2, was involved. We propose that CP can be a new therapeutic formula against ALI.

Induction of early meconium evacuation promotes feeding tolerance in very low birth weight infants.

BACKGROUND: A delay in reaching full enteral feeding is linked to poorer outcome in preterm neonates. Meconium retention has been viewed as a cause of bowel dysfunction in very low birth weight infants (VLBWI). Thus, adequate evacuation of meconium could help to promote feeding tolerance. OBJECTIVES: Our goal was to determine the effect of the induction of early meconium evacuation on feeding tolerance in VLBWI. METHODS: An observational study involving two subsequent periods was performed in inborn infants with birth weights of <1,500 g, before (control) and after (study) the induction of early meconium evacuation by routine glycerin enema. The total duration of these periods was from January 2003 to December 2005. To evaluate feeding tolerance, we measured time to achieve full enteral feeding. Complications such as sepsis and necrotizing enterocolitis were compared. RESULTS: The study group achieved full enteral feeding significantly faster than the control group (hazard ratio (HR) = 2.9; 95% confidence interval (CI) = 1.8-4.8), and this effect was more definite in infants with a birth weight of <1,000 g (HR = 4.6; 95% CI = 1.9-11.1). The study group passed first meconium faster than the control group (median = 1.4 vs. 3.7 days; p < 0.001). Sepsis, especially as determined by positive culture in central venouscatheter, was significantly reduced in the study group (7.7 vs. 27.8%; p = 0.02). CONCLUSIONS: The induction of early meconium evacuation had a significantly positive effect on feeding tolerance and sepsis prevention in VLBWI.