Antiviral activity of porcine interferon omega 7 against foot-and-mouth disease virus in vitro.

Foot-and-mouth disease (FMD) is a disease of worldwide economic importance, and vaccines play an important role in preventing FMDV outbreaks. However, new control strategies are still needed since FMDV outbreaks still occur in some disease-free countries. Currently, interferon (IFN)-based strategies have been demonstrated to be an efficient biotherapeutic option against FMDV; however, interferon omega (IFN-ω) has not yet been assessed in this capacity. Thus, this study evaluated the antiviral activity of porcine IFN omega 7 (PoIFN-ω7) against FMDV. After the PoIFN-ω7 was expressed and purified, cell proliferation assays and quantitative real-time reverse transciption-polymerase chain reaction were used to evaluate the effective anti-cytopathic concentration of PoIFN-ω7 and its effectiveness pre- and post-infection with FMDV in swine kidney cells (IBRS-2). Results showed the rHis-PoIFN-ω7 fusion protein was considerably expressed using Escherichia coli BL21 (DE3) strain, and the recombinant protein exhibited significant in vitro protection against FMDV, including two strains belonging to type O and A FMDV, respectively. In addition, PoIFN-ω7 upregulated the transcription of Mx1, ISG15, OAS1, and PKR genes. These findings indicated that IFN-ω has the potential for serving as a useful therapeutic agent to prevent FMDV or other viral outbreaks in pigs.

Inhibitory effects of homoharringtonine on foot and mouth disease virus in vitro.

Foot-and-mouth disease (FMD) is a highly contagious disease that affects cloven-hoof animals including cattle, swine, sheep, goats, and lots of wild species. Effectively control measures are urged needed. Here, we showed that homoharringtonine treatment exhibited a strong inhibitory effect against two different strains of FMDVs (O/MYA98/BY/2010 and A/GD/MM/2013) in swine kidney (IBRS-2) cells. Further experiments demonstrated that homoharringtonine did not affect virus attachment or entry. Using time-of-addition assays, we found that the antiviral activity of homoharringtonine occurred primarily during the early stage of infection. These results demonstrated that homoharringtonine might be an effective anti-FMDV drug. Further studies are required to explore the antiviral activity of homoharringtonine against FMDV replication in vivo.

Antiviral effect of amiloride on replication of foot and mouth disease virus in cell culture.

Recently, amiloride was shown to potently suppress Coxsackievirus B3 (CVB3) replication. In the current study, we investigated whether amiloride could also exhibit antiviral activity against foot-and-mouth disease virus (FMDV), which belongs to the same family (Picornaviridae) as CVB3. We found that amiloride exerted antiviral activity in a dose-dependent manner against two strains of FMDV in IBRS-2 cells, with slight cytotoxicity at 1000 µM. Besides, amiloride did not inhibit the attachment and entry of FMDV in IBRS-2 cells, but prevented early viral replication. These data implied that amiloride could be a promising candidate for further research as a potential antiviral drug against FMDV infection.

A novel type I interferon, interferon alphaomega, shows antiviral activity against foot-and-mouth disease virus in vitro.

Recently, a novel type I interferon alphaomega (IFN-αω), also known as IFN-µ, was identified. However, the biological activity of IFN-αω remain poorly understood. In this study, the porcine IFN-αω (PoIFN-αω) was expressed, purified, and its antiviral activities assessed by its ability to inhibit the cytopathic effect caused by FMDV on IBRS-2 cells. In addition, q-PCR was used to evaluate the expression of IFN-stimulated genes induced by PoIFN-αω. It was found that PoIFN-αω exerted effective antiviral activity against FMDV pre- and post-infection. Additionally, PoIFN-αω induced the transcription of IFN-stimulated genes, including Mx1, ISG15, OAS1, and PKR genes. Our study reported a new indication of PoIFN-αω as an effective anti-FMDV agent for the first time.

In Vitro and in Vivo Antiviral Activity of Mizoribine Against Foot-And-Mouth Disease Virus.

Foot-and-mouth disease (FMD) is a highly contagious viral disease of cloven-hoofed animals, which has significant economic consequences in affected countries. As the currently available vaccines against FMD provide no protection until 4-7 days post-vaccination, the only alternative method to control the spread of FMD virus (FMDV) during outbreaks is the application of antiviral agents. Hence, it is important to identify effective antiviral agents against FMDV infection. In this study, we found that mizoribine has potent antiviral activity against FMDV replication in IBRS-2 cells. A time-of-drug-addition assay demonstrated that mizoribine functions at the early stage of replication. Moreover, mizoribine also showed antiviral effect on FMDV in vivo. In summary, these results revealed that mizoribine could be a potential antiviral drug against FMDV.

Genetic and environmental factors contribute to variation in cell wall composition in mature desi chickpea (Cicer arietinum L.) cotyledons.

Chickpea (Cicer arietinum L.) is an important nutritionally rich legume crop that is consumed worldwide. Prior to cooking, desi chickpea seeds are most often dehulled and cleaved to release the split cotyledons, referred to as dhal. Compositional variation between desi genotypes has a significant impact on nutritional quality and downstream processing, and this has been investigated mainly in terms of starch and protein content. Studies in pulses such as bean and lupin have also implicated cell wall polysaccharides in cooking time variation, but the underlying relationship between desi chickpea cotyledon composition and cooking performance remains unclear. Here, we utilized a variety of chemical and immunohistological assays to examine details of polysaccharide composition, structure, abundance, and location within the desi chickpea cotyledon. Pectic polysaccharides were the most abundant cell wall components, and differences in monosaccharide and glycosidic linkage content suggest both environmental and genetic factors contribute to cotyledon composition. Genotype-specific differences were identified in arabinan structure, pectin methylesterification, and calcium-mediated pectin dimerization. These differences were replicated in distinct field sites and suggest a potentially important role for cell wall polysaccharides and their underlying regulatory machinery in the control of cooking time in chickpea.

Lithium chloride inhibits early stages of foot-and-mouth disease virus (FMDV) replication in vitro.

Foot-and-mouth disease virus (FMDV) causes an economically important and highly contagious disease of cloven-hoofed animals such as cattle, swine, and sheep. FMD vaccine is the traditional way to protect against the disease, which can greatly reduce its occurrence. However, the use of FMD vaccines to protect early infection is limited. Therefore, the alternative strategy of applying antiviral agents is required to control the spread of FMDV in outbreak situations. As previously reported, LiCl has obviously inhibition effects on a variety of viruses such as transmissible gastroenteritis virus (TGEV), infectious bronchitis coronavirus (IBV), and pseudorabies herpesvirus and EV-A71 virus. In this study, our findings were the first to demonstrate that LiCl inhibition of the FMDV replication. In this study, BHK-21 cell was dose-dependent with LiCl at various stages of FMDV. Virus titration assay was calculated by the 50% tissue culture infected dose (TCID50 ) with the Reed and Muench method. The cytotoxicity assay of LiCl was performed by the CCK8 kit. The expression level of viral mRNA was measured by RT-qPCR. The results revealed LiCl can inhibit FMDV replication, but it cannot affect FMDV attachment stage and entry stage in the course of FMDV life cycle. Further studies confirmed that the LiCl affect the replication stage of FMDV, especially the early stages of FMDV replication. So LiCl has potential as an effective anti-FMDV drug. Therefore, LiCl may be an effective drug for the control of FMDV. Based on that, the mechanism of the antiviral effect of LiCl on FMDV infection is need to in-depth research in vivo.

Web thickness determines the therapeutic effect of endoscopic keel placement on anterior glottic web.

This work is a retrospective analysis to investigate the critical risk factor for the therapeutic effect of endoscopic keel placement on anterior glottic web. Altogether, 36 patients with anterior glottic web undergoing endoscopic lysis and silicone keel placement were enrolled. Their voice qualities were evaluated using the voice handicap index-10 (VHI-10) questionnaire, and improved significantly 3 months after surgery (21.53 ± 3.89 vs 9.81 ± 6.68, P < 0.0001). However, 10 (27.8%) cases had web recurrence during the at least 1-year follow-up. Therefore, patients were classified according to the Cohen classification or web thickness, and the recurrence rates were compared. The distribution of recurrence rates for Cohen type 1 ~ 4 were 28.6, 16.7, 33.3, and 40%, respectively. The difference was not statistically significant (P = 0.461). When classified by web thickness, only 2 of 27 (7.41%) thin type cases relapsed whereas 8 of 9 (88.9%) cases in the thick group reformed webs (P < 0.001). These results suggest that the therapeutic outcome of endoscopic keel placement mostly depends on the web thickness rather than the Cohen grades. Endoscopic lysis and keel placement is only effective for cases with thin glottic webs. Patients with thick webs should be treated by other means.

Differences in glycosyltransferase family 61 accompany variation in seed coat mucilage composition in Plantago spp.

Xylans are the most abundant non-cellulosic polysaccharide found in plant cell walls. A diverse range of xylan structures influence tissue function during growth and development. Despite the abundance of xylans in nature, details of the genes and biochemical pathways controlling their biosynthesis are lacking. In this study we have utilized natural variation within the Plantago genus to examine variation in heteroxylan composition and structure in seed coat mucilage. Compositional assays were combined with analysis of the glycosyltransferase family 61 (GT61) family during seed coat development, with the aim of identifying GT61 sequences participating in xylan backbone substitution. The results reveal natural variation in heteroxylan content and structure, particularly in P. ovata and P. cunninghamii, species which show a similar amount of heteroxylan but different backbone substitution profiles. Analysis of the GT61 family identified specific sequences co-expressed with IRREGULAR XYLEM 10 genes, which encode putative xylan synthases, revealing a close temporal association between xylan synthesis and substitution. Moreover, in P. ovata, several abundant GT61 sequences appear to lack orthologues in P. cunninghamii. Our results indicate that natural variation in Plantago species can be exploited to reveal novel details of seed coat development and polysaccharide biosynthetic pathways.

Short-Term Dynamic Observation of the Color Change and Enhancement Effect of Polyethylenimine-Entrapped Gold Nanoparticles Used for Indirect Lymphography.

AIMS: To dynamically observe the color change and enhancement effect of gadolinium-loaded polyethylenimine-entrapped gold nanoparticles (Gd-Au PENPs) as a dual-mode CT/MRI contrast agent for sentinel lymph node (SLN) identification. METHODS: In 6 rabbits, Gd-Au PENPs were injected into the right side tongue submucosa, after which the color change of cervical draining lymph nodes was observed. Then the draining lymph nodes were examined by CT/MRI scan. Another 6 rabbits were randomly divided into a CT lymphography (CT-LG) and a MRI lymphography (MRI-LG) group and examined by CT/MRI scan 1 and 30 min after injection. Then SLNs were identified under the guidance of CT/MRI-LG. RESULTS: The ipsilateral afferent lymphatic vessels, cervical draining lymph nodes and efferent lymphatic vessels were stained black orderly and constantly after Gd-Au PENPs injection. Thirty minutes after injection, all cervical lymph nodes were excised to be examined by CT/MRI scan, and only the black-stained draining lymph nodes were enhanced. The 6 rabbits with CT/MRI-LG showed clear enhancement of SLNs. With indirect lymphography, the black-stained SLNs were easily visualized at autopsy. In all rabbits, the identification rate of lingual SLNs was 100%. CONCLUSIONS: Rabbit lingual SLNs could be identified with indirect lymphography using Gd-Au PENPs as a dual-mode contrast agent.

Effects of magnesium-substituted nanohydroxyapatite coating on implant osseointegration.

OBJECTIVE: The objective of this study was to compare magnesium-substituted and pure hydroxyapatite coatings on the promotion of osteogenesis in vitro and on the osseointegration in vivo. METHODS: Electrochemically deposited pure hydroxyapatite (EDHA) or electrochemically deposited magnesium-substituted hydroxyapatite (EDMHA) coatings were formed on the surface of pure titanium disks or implants. MC3T3-E1 preosteoblasts were cultured in the EDHA and EDMHA coated disks, and cell growth, alkaline phosphatase (ALP) activity, and osteocalcin secretion were measured at various time points. For studies on osseointegration, 30 roughened implants coated either with EDHA or EDMHA (n = 15 for each coating) were implanted in the femurs of 15 NZW rabbits. After 2, 4, and 8 weeks, femurs were retrieved and prepared for histomorphometric evaluation (n = 5 for each coating at each time point). RESULTS: MC3T3-E1 cells cultured on EDMHA coated disks showed increased cell number, ALP, and osteocalcin secretion compared with the EDHA coated disks at all time points (P < 0.05 for all). Histologic observation of the coated implants showed woven bone in direct contact with both implant surfaces after 2 weeks and mature bone after 8 weeks. While there were no differences in the amount of bone between the threads at any time point, the percentage of implant in direct contact with bone (bone implant contact) was slightly higher along the EDMHA coated implants at 2 weeks (P = 0.086), although this difference was no longer seen at 4 and 8 weeks. CONCLUSION: Mg-substituted HA coated surfaces promote osteogenic differentiation of preosteoblasts in vitro and may improve implant osseointegration during the early stages of bone healing compared with pure EDHA coated surfaces.

Bone response to the multilayer BMP-2 gene coated porous titanium implant surface.

OBJECTIVES: Evaluate hBMP-2 expression following gene delivery from plasmid multilayers formed on sandblasted titanium in vitro and bone formation around similarly prepared implant surfaces in vivo. MATERIALS AND METHODS: Multilayers of cationic lipid/rhBMP-2 plasmid DNA complex (LDc) and anionic hyaluronic acid (HA) was assembled on sandblasted-dual acid etched pure titanium disks or implant surfaces using layer-by-layer (LBL) assembly. Gene delivery and hBMP-2 expression in cells exposed to the LDc multilayers was measured in vitro. To determine the effect of BMP delivery from such multilyaers in vivo, roughened implants coated with BMP-2 LDc multilayers or uncoated control implants (n = 15 for both) were implanted in the femurs of NZW rabbits. After 2, 4, 8 weeks, femurs were retrieved and prepared for histomorphometric evaluation (n = 5 rabbits per time point). RESULTS: MC3T3-E1 cells cultured directly on the BMP-2 LDc coated titanium disks showed EGFP and hBMP-2 expression after 48 h in culture. Increased gene delivery occurred by increasing the number of assembly layers when cells were cultured for 48 h. Cells cultured on LDc coated surfaces had significantly higher cell viability than control cells cultured on uncoated porous titanium surfaces. Histologic observation of the implants showed that after 4 weeks healing, the bone to implant contact (BIC) on the LDc coated surface was much lower than that on the control surface, but didn't reach significant. In contrast, the percentage of bone within the implant's threads was significantly higher than the control group (P = 0.047). CONCLUSION: The BMP-2 gene coated sandblasted dual acid etched titanium implants slightly accelerated early bone formation around implants.

[Application of CAD/CAM technique in three-dimensional reconstruction of zygomatic complex defect].

OBJECTIVE: To evaluate the efficacy of computer-aided design and computer-aided manufacturing (CAD/CAM) technique in reconstruction of zygomatic complex defect. METHODS: Three-dimensional computed tomography (3D-CT) and rapid prototyping (RP) techniques were applied to produce facsimile models in 13 patients with zygomatic complex defects resulted from trauma or tumor ablation. The zygomatic complex defect was reconstructed using a mirroring technique by CAD/CAM. Titanium mesh was prefabricated on individual facsimile models and combined with autologous bone graft and pedicled buccal fat pad flap for reconstruction. RESULTS: The measuring data of zygomatic complex defect between facsimile model and intraoperative findings was extremely consistent. The prefabrication of reconstructive titanium mesh was matched with zygomatic complex defect in the surgery. The postoperative 3D-CT image demonstrated the symmetry of reconstructed zygomatic complex. The functional recovery and facial contours were satisfactory in all cases. CONCLUSION: The application of CAD/CAM technique can simulate the surgery procedures accurately, which contributes to shorten the actual operative time. RP techniques can reconstruct the facial contours symmetry and recover the function of zygomatic complex defect.

[Construction of a multiple-scale implant surface with super-hydrophilicity].

OBJECTIVE: To construct a multiple-scale organized implant surface with super-hydrophilicity. METHODS: The SiC paper polished titanium disc was sandblasted and treated with HF/HNO3 and HCl/H2SO4, then acid-etched with H2SO4/H2O2. The physicochemical properties of the surfaces were characterized by scanning electron microscope, static state contact angle and X-ray diffraction. MC3T3-E1 cells were used to evaluate the effects of the surface on the cell adhesion, proliferation and differentiation. RESULTS: The acid-etching process with a mixture of H2SO4/H2O2 superimposed the nano-scale structure on the micro-scale texture. The multiple-scale implant surface promoted its hydrophilicity and was more favorable to the responses of osteoprogenitor cells, characterized by increased DNA content, enhanced ALP activity and promoted OC production. CONCLUSION: A multiple-scale implant surface with super-hydrophilicity has been constructed in this study, which facilitates cell proliferation and adhesion.

Mechanical and histomorphometric evaluations of rough titanium implants treated with hydrofluoric acid/nitric acid solution in rabbit tibia.

PURPOSE: The aim of this study was to investigate the bone response to rough titanium implants treated with hydrofluoric acid/nitric acid (HF/HNO3) solution. MATERIALS AND METHODS: Implants were treated with HF/HNO3 solution (test implants) or without HF/HNO3 solution (control implants). Forty-five test and 45 control implants were inserted into both tibias of 15 rabbits. After 2, 4, and 8 weeks in situ, tibias were retrieved and prepared for removal torque testing and histomorphometric evaluation. The removed implants were prepared and observed with an electron microscope. RESULTS: Mechanical tests showed that mean removal torque values for the test implants were higher than those of the control implants after 8 weeks (33.1 Ncm versus 25.7 Ncm, P = .012). Histomorphometric analysis showed that the bone area in the threads of the cortical bone region was significantly higher for test implants (81.99% and 86.38%) than for control implants (75.33% and 81.62%) after 4 and 8 weeks of healing, respectively. The implant-bone contact rate in the cortical region was higher for test implants than for control implants after 8 weeks in situ (79.56% versus 68.45%, P = .003). CONCLUSIONS: The treatment with HF/HNO3 solution promotes bone formation and osseointegration after 4 and 8 weeks of bone healing in the rabbit tibia model.

Hyperbranched anion exchangers prepared from polyethylene polyamine modified polymeric substrates for ion chromatography.

High hydrophilic anion stationary phases play a crucial role in the separation behavior of ion chromatography. Herein, we report novel polymeric anion exchangers grafted with polyethylene polyamines, including ethylenediamine, diethylenetriamine, triethylenetetramine and tetraethylene pentaamine, via a facile epoxy-amine polymerization method. The anion exchangers were characterized by scanning electron microscopy, thermogravimetry, Fourier transform infrared spectrometry and elemental analysis. The chromatographic performance of the stationary phases was evaluated with the separation of common inorganic anions, organic weak acids and highly polarizable anions. Seven common anions (F-, Cl-, NO2-, Br-, NO3-, SO42- and HPO42-) can be separated within 18 min by using hydroxide eluent in isocratic mode. By adopting different polyethylene polyamines as hyperbranched units, the four types of new stationary phases displayed high efficiencies and good reproducibility. The columns exhibit large exchange capacities at 76.5-184.8 µmol•column-1 (4.6 × 150 mm, i.d.) with efficiency up to 20293 plate m-1 (Cl-). The RSDs of the retention time were less than 0.27% and the RSDs of the efficiency were less than 1.95% by consecutive injections after working for two months. The self-fabricated column was successfully applied to determine the chloride content in exhaled breath condensate.

In vivo comparison of bone formation on titanium implant surfaces coated with biomimetically deposited calcium phosphate or electrochemically deposited hydroxyapatite.

PURPOSE: The purpose of this study was to investigate and compare bone formation on titanium implant surfaces coated with biomimetically deposited calcium phosphate (BDCaP) or electrochemically deposited hydroxyapatite (EDHA). MATERIALS AND METHODS: The implants were separated into three groups: a control group, a BDCaP group, and an EDHA group. Surface analysis was performed by field-emission scanning electron microscopy, x-ray diffractometry, and Fourier transform infrared spectroscopy. Implants were inserted in a randomized arrangement into rabbit tibiae. After 2, 4, and 8 weeks, the tibiae were retrieved and prepared for histomorphometric evaluation. RESULTS: Field-emission scanning electron microscopy showed that the BDCaP crystals were flakelike and the EDHA crystals were rodlike with a hexagonal cross section. X-ray diffractometric patterns and Fourier transform infrared spectroscopy spectra showed that the BDCaP coating consisted of HA and octacalcium phosphate, whereas the EDHA coating consisted of HA. Histologic observation showed that new bone on the EDHA-coated implant became mature after 4 weeks, while new bone on the control and BDCaP-coated implants was mature after 8 weeks. The EDHA implant showed significantly greater BIC and bone area compared to the control and BDCaP implants during 4 to 8 weeks. The BDCaP coating failed to show increased bone formation during the test period. CONCLUSION: The present EDHA coating has good bone formation properties, while the BDCaP coating has weaker bone formation properties.

Cell response of titanium implant with a roughened surface containing titanium hydride: an in vitro study.

PURPOSE: The purpose of this study was to investigate the effect of surface chemistry of a sandblasted and acid-etched implant (with and without titanium hydride [TiH(2)]) on cell attachment, proliferation, and differentiation of preosteoblasts (MC3T3-E1). MATERIALS AND METHODS: Sandblasted and dual acid-etched titanium discs comprised the test group, whereas sandblasted, acid-etched, and heat-treated discs comprised the control group. Both groups' discs were sent for surface characterization. MC3T3-E1 cells were cultured on these 2 groups' discs, and then cell attachment, cell proliferation, and cell differentiation were analyzed. RESULTS: Scanning electron microscope analysis showed that the titanium discs in the 2 groups shared the same surface topography; however, x-ray diffraction examination showed that the TiH(2) diffractions only appeared in the test group. Cell attachment and cell proliferation were much better in the test group than in the control group at all time points investigated (P < .05). The expressions of alkaline phosphatase and osteocalcin were significantly higher in the test group than in the control group for both protein and transcription level at every time point (P < .05 or P < .01). CONCLUSIONS: These results suggested that surface chemistry played a significant role in cell response to the sandblasted and acid-etched surface and the presence of TiH(2) might promote the attachment, proliferation, and differentiation of preosteoblasts.

Biomechanical comparison of biomimetically and electrochemically deposited hydroxyapatite-coated porous titanium implants.

PURPOSE: The purpose of this study was to investigate the effects of biomimetically and electrochemically deposited hydroxyapatite on the fixation of an implant with bone tissue. MATERIALS AND METHODS: Implants were separated into 3 groups: roughened group, biomimetically deposited calcium-phosphorus (BDCaP) group, and electrochemically deposited hydroxyapatite (EDHA) group. We randomly inserted 90 implants into the femurs of 45 rabbits. After 2, 4, and 8 weeks, the femurs were retrieved and prepared for removal torque tests (RTQs) and field-emission scanning electron microscopy observation. RESULTS: During the test period, the EDHA group showed significantly greater RTQ values than did the roughened group and BDCaP group. The BDCaP group failed to increase the RTQ values compared with the roughened group. Field-emission scanning electron microscopy observation showed that the amount of attached bone tissue on the EDHA-coated implant surface was more than that on the roughened and BDCaP-coated implant surfaces during the test period. CONCLUSION: The electrochemical hydroxyapatite coating contributes to the fixation between bone and implant compared with the roughened surface, whereas the biomimetic calcium-phosphorus coating has little effect on the fixation.

[Preparation of brain targeted immunoliposomes].

To prepare a kind of effective non-viral transduction vector, which can deliver exogenous gene into the brain, this vector can be injected through vein system and has the ability to penetrate blood brain barrier. Several groups of materials proportion, type of oil phase, water-oil ratio, phosphatides-cholesterol ratio, temperature of steaming, ultrasonic temperature and time were compared for optimization. Well-constructed immunoliposomes encapsuling LacZ gene were infused into rats through tail vein. 48 h after injection, expression product beta-galactosidase of LacZ gene was detected by histochemistry staining to convince the validity of immunoliposomes as non-viral vectors. The best proportion of synthesis immunoliposomes is as following: phosphatides-cholesterol ratio is 1:1, lipids/drug is 100:1, the type of oil phrase is dichloromethane, oil-water ratio is 4:1, temperature of steaming is 30 degrees C, ultrasonic temperature and time is 10 degrees C and 5 min. At last, 10% trehalose was added as a stabilizer. The entrapment rate is 87.24% and antibody coupling rate is 69%. When immunoliposomes were infused into rats, the expression of LacZ gene could be observed in the brain and periphery organs. Through the best proportion of materials, gene delivering immunoliposomes had been synthesized successfully. This non-viral vector can deliver exogenous gene penetrating blood brain barrier and express in the brain, and will be well-used in the field of gene therapy of cerebral diseases.