Comparative antiplatelet and antithrombotic effects of red ginseng and fermented red ginseng extracts.

Background: Fermentation may alter the bioavailability of certain compounds, which may affect their efficacy and pharmacological responses. This study investigated the antiplatelet effects of red ginseng extract (RGE) and fermented red ginseng extract (FRG). Methods: A rodent model was used to evaluate the antiplatelet and antithrombotic effects of the extracts. Rats were orally fed with human equivalent doses of the extracts for 1 week and examined for various signaling pathways using standard in vivo and ex vivo techniques. Light transmission aggregometry was performed, and calcium mobilization, dense granule secretion, integrin αIIbß3-mediated signaling molecules, cyclic nucleotide signaling events, and various protein molecules were evaluated ex vivo in collagen-stimulated washed platelets. Furthermore, antithrombotic properties were evaluated using a standard acute pulmonary thromboembolism model, and the effects on hemostasis were investigated using rat and mice models. Results: Both RGE and FRG significantly inhibited platelet aggregation, calcium mobilization, and dense granule secretion along with integrin-mediated fibrinogen binding and fibrinogen adhesion. cAMP levels were found to be elevated in RGE-treated rat platelets. Ginseng extracts did not exert any effect on prothrombin time and activated partial thromboplastin time. RGE-treated mice showed significantly better survival under thrombosis than FRG-treated mice, with no effects on hemostasis, whereas FRG-treated mice exhibited a slight increment in bleeding time. Conclusion: Both extracts, especially RGE, are remarkable supplements to maintain cardiovascular health and are potential candidates for the treatment and prevention of platelet-related cardiovascular disorders.

Trifolium pratense ethanolic extract alters the gut microbiota composition and regulates serum lipid profile in the ovariectomized rats.

BACKGROUND: Trifolium pratense (red clover) ethanolic extract (TPEE) has been used as a popular over-the-counter remedy for the management of menopausal symptoms. Prolonged consumption of herbal extract has been shown to regulate the composition of gut microbiota. This study was designed to elucidate the influence of TPEE on the gut microbiota composition in the ovariectomized (OVX) rats. METHODS: OVX rats were treated with TPEE at 125, 250, 500 mg/kg/day, or controls (pomegranate extract, 500 mg/kg/day; estradiol, 25 µg/kg/day) for 12 weeks. Gut microbiota analysis was conducted by extracting the microbial DNA from fecal samples and microbiome taxonomic profiling was carried out by using next-generation sequencing. The levels of serum biomarkers were analyzed using enzyme-linked immunosorbent assay (ELISA) kit. The prediction of functional biomarker of microbiota was performed using PICRUSt to investigate the potential pathways associated with gut health and serum lipid profile regulation. To study the correlation between gut microbiota composition and serum lipid levels, Spearman's correlation coefficients were defined and analyzed. Additionally, gas chromatography-mass spectrometry analysis was conducted to uncover additional physiologically active ingredients. RESULTS: TPEE-treated OVX rats showed significant reduction in serum triglycerides (TG), total cholesterols (TCHOL), and LDL/VLDL levels but increase in HDL level. The alteration in the pathways involve in metabolism was the most common among the other KEGG categories. Particularly, TPEE also significantly reduced the relative abundance of sequences read associated with inflammatory bowel disease (IBD) and the peroxisome proliferator-activated receptor (PPAR) signalling pathway. TPEE intervention was seen to reduce the Firmicutes to Bacteroidetes (F/B) ratio in the OVX rats, denoting a reduction in microbial dysbiosis in the OVX rats. Correlation analysis at the phylum level revealed that Bacteriodetes and Proteobacteria were strongly correlated with serum TG, TCHOL and HDL levels. At the species level, Bifidobacterium pseudolongum group was seen to positively correlate with serum HDL level and negatively correlated with serum AST, ALT, LDL/VLDL, TCHOL, and TG levels. CONCLUSIONS: TPEE treatment showed therapeutic benefits by improving the intestinal microbiota composition which strongly correlated with the serum lipid and cholesterol levels in the OVX rats.

Modulation of quorum sensing-controlled virulence factors by Nymphaea tetragona (water lily) extract.

ETHNOPHARMACOLOGICAL RELEVANCE: Nymphaea tetragona is a widely distributed ornamental species with ethnomedicinal uses in the treatment of diarrhea, dysentery, eruptive fevers, and infections. The anti-infectious activities of this herb have already been assessed to clarify its traditional use as a medicine. AIM OF STUDY: In this study, we aimed to verify the inhibitory effects of N. tetragona 50% methanol extract (NTME) on quorum sensing (QS)-controlled virulence factors of bacteria since QS and its virulence factors are novel targets for antimicrobial therapy. MATERIALS AND METHODS: The antibacterial activity of this extract was evaluated against Chromobacterium violaceum and Pseudomonas aeruginosa. The inhibition of the violacein pigment of C. violaceum by NTME was determined qualitative and quantitative using standard methods. The effects of NTME on swarming motility, biofilm viability, pyocyanin production, and LasA protease activity were evaluated using P. aeruginosa. Finally, the in vitro and in vivo cytotoxicity of NTME were verified by MTT assay and oral administration to rats, respectively. RESULTS: The extract had concentration-dependent antibacterial activity against gram-negative bacteria. NTME at 1/2× minimum inhibitory concentration (MIC), 1× MIC and 2× MIC significantly lowered the levels of violacein of C. violaceum compared to that of the control. The swarming motility of P. aeruginosa was inhibited by ≥70% by treatment with 1/2× MIC of NTME. There were remarkable reductions in pyocyanin production and LasA protease activity in the overnight culture supernatant of P. aeruginosa supplemented with NTME when compared with that of the untreated control. The confocal micrographs of 24h biofilms of P. aeruginosa exposed to NTME exhibited a lower number of live cells than the control. No toxic effect was observed in in vitro and in vivo cytotoxicity assays of NTME. CONCLUSIONS: NTME was demonstrated to have significant concentration-dependent inhibitory effects on quorum sensing-mediated virulence factors of bacteria with non-toxic properties, and could thus be a prospective quorum sensing inhibitor.

Establishment of optimal scan delay for multi-phase computed tomography using bolus-tracking technique in canine pancreas.

To establish a protocol for a multi-phase computed tomography (CT) of the canine pancreas using the bolus-tracking technique, dynamic scan and multi-phase CT were performed in six normal beagle dogs. The dynamic scan was performed for 60 sec at 1-sec intervals after the injection (4 ml/sec) of a contrast medium, and intervals from aortic enhancement appearance to aortic, pancreatic parenchymal and portal vein peaks were measured. The multi-phase CT with 3 phases was performed three times using a bolus-tracking technique. Scan delays were 0, 15 and 30 in first multi-phase scan; 5, 20 and 35 in second multi-phase scan; and 10, 25 and 40 sec in third multi-phase scan, respectively. Attenuation values and contrast enhancement pattern were analyzed from the aorta, pancreas and portal vein. The intervals from aortic enhancement appearance to aortic, pancreatic parenchymal and portal vein peaks were 3.8 ± 0.7, 8.7 ± 0.9 and 13.3 ± 1.5 sec, respectively. The maximum attenuation values of the aorta, pancreatic parenchyma and portal vein were present at scan sections with no scan delay, a 5-sec delay and a 10-sec delay, respectively. When a multi-phase CT of the canine pancreas is triggered at aortic enhancement appearance using a bolus-tracking technique, the recommended optimal delay times of the arterial and pancreatic parenchymal phases are no scan delay and 5 sec, respectively.

Korean Red Ginseng saponin fraction modulates radiation effects on lipopolysaccharide-stimulated nitric oxide production in RAW264.7 macrophage cells.

BACKGROUND: In previous work, we reported that Korean Red Ginseng saponin fraction (RGSF) showed anti-inflammatory activities in vitro and in vivo. METHODS: The present study investigated the radioprotective properties of RGSF by examining its effects on ionizing radiation (IR)-enhanced and lipopolysaccharide (LPS)-mediated inflammatory responses in murine macrophage cells. RESULTS: RGSF induced strong downregulation of IR-enhanced and LPS-induced proinflammatory responses such as nitric oxide (NO) production (Inhibitory Concentration 50 (IC50) = 5.1 ± 0.8 µM) and interleukin-1ß levels. RGSF was found to exert its radioprotective effects by inhibition of a signaling cascade that activated checkpoint kinase 2-nuclear factor-κB. In addition, RGSF strongly inhibited IR-enhanced LPS-induced expression of hemoxyganase-1, implying that the latter may be a potential target of RGSF. CONCLUSION: Taken together, our data suggest that RGSF can be considered and developed for use as an effective radioprotective agent with minimal adverse effects.

Use of contrast-enhanced CT in the diagnosis of abscesses in cattle.

We report here the non-contrast and contrast-enhanced computed tomography performed in two calves with brain abscess and multiple pulmonary abscesses with pharyngeal abscess, respectively. Contrast-enhanced computed tomography was useful in the diagnosis of these diseases in both calves. The diseases were confirmed by histopathological examination.

Virtopsy in a red kangaroo with oral osteomyelitis.

This report describes the use of computed tomography (CT) in a nondomestic species. Postmortem CT was performed on a red kangaroo (Macropus rufus) and a diagnosis of oral osteomyelitis was made. CT examination revealed bony remodeling of the right mandible, an intraosseous lesion of the right temporal bone, muscle necrosis around the right mandible, and the absence of the right, first, upper molar tooth. Cardiac and intrahepatic gas and a distended intestine due to postmortem gas accumulation were also seen. All the lesions identified with CT were also identified by conventional necropsy, except the cardiac and intrahepatic gases. Virtopsy may be a useful procedure for the noninvasive identification of cause of death and as a guide for necropsy in animals.

Measurement of canine pancreatic perfusion using dynamic computed tomography: influence of input-output vessels on deconvolution and maximum slope methods.

OBJECTIVE: We investigated whether the prerequisite of the maximum slope and deconvolution methods are satisfied in pancreatic perfusion CT and whether the measured parameters between these algorithms are correlated. METHODS: We examined nine beagles injected with iohexol (200 mgI kg(-1)) at 5.0 ml s(-1). The abdominal aorta and splenic and celiac arteries were selected as the input arteries and the splenic vein, the output veins. For the maximum slope method, we determined the arterial contrast volume of each artery by measuring the area under the curve (AUC) and compared the peak enhancement time in the pancreas with the contrast appearance time in the splenic vein. For the deconvolution method, the artery-to-vein collection rate of contrast medium was calculated. We calculated the pancreatic tissue blood flow (TBF), tissue blood volume (TBV), and mean transit time (MTT) using both algorithms and investigated their correlation based on vessel selection. RESULTS: The artery AUC significantly decreased as it neared the pancreas (P<0.01). In all cases, the peak time of the pancreas (11.5±1.6) was shorter than the appearance time (14.1±1.6) in the splenic vein. The splenic artery-vein combination exhibited the highest collection rate (91.1%) and was the only combination that was significantly correlated between TBF, TBV, and MTT in both algorithms. CONCLUSION: Selection of a vessel nearest to the pancreas is considered as a more appropriate prerequisite. Therefore, vessel selection is important in comparison of the semi-quantitative parameters obtained by different algorithms.

The relationship between the size of caudolateral curvilinear osteophyte of the canine femoral neck and the radiographic view.

Caudolateral curvilinear osteophyte (CCO), an osteophyte at the site of joint capsule attachment on the caudal aspect of the femoral neck, has been advocated as a radiographic criterion for coxofemoral subluxation. The correlation between the presence of CCO on radiographs (radiographic-CCO), the size of the CCO (CCO index) on three-dimensional computed tomographic (CT) images, and hip evaluation using transverse CT images was assessed in 22 Border Collies. CCOs were detected on the radiographs and CT images of 32% and 100% femurs, respectively. The CCO index correlated significantly with radiographic-CCO, but a large CCO index did not necessarily imply that the CCO was visible on radiographs. Hence, radiographic-CCO findings should be used cautiously in hip evaluation of Border Collies.

Computed tomographic evaluation of cerebral ventricular size in clinically normal calves.

OBJECTIVE: To evaluate lateral ventricular size in clinically normal calves by use of computed tomography and to examine the relationships between ventricular height (Vh), ventricular area (VA), and ventricular volume (VV). ANIMALS: 14 Holstein calves. Procedures-14 calves underwent computed tomography of the head with transverse images acquired from the rostral aspect of the frontal lobe continuing caudally to the level of the foramen magnum. Hemispheric height, Vh, VA, and hemispheric area were measured on images obtained at the level of the interventricular foramen. Ventricular volume was calculated by multiplying the sum of VAs measured on each transverse image by the total slice thickness. The left Vh-to-right Vh ratio was calculated to determine the degree of ventricular asymmetry, which was categorized as normal (ie, symmetric) to minimally asymmetric, mildly asymmetric, or severely asymmetric. RESULTS: Mean +/- SD values for Vh and the Vh-to-hemispheric height ratio were 4.96 +/- 1.56 mm and 7.47%, respectively. The mean VA was 114.29 +/- 47.68 mm(2), and the mean VV was 2,443.50 +/- 1,351.50 mm3. Normal to minimally asymmetric ventricles were identified in 13 calves, and mildly asymmetric ventricles were identified in 1 calf. Significant correlations were found between Vh and VA and between Vh and VV. CONCLUSIONS AND CLINICAL RELEVANCE: These results establish reference values for ventricular size in clinically normal calves and suggest that Vh measurement may be a simple and useful technique for examining size of the cerebral ventricles in calves.

Influence of osmolarity of contrast medium and saline flush on computed tomography angiography: comparison of monomeric and dimeric iodinated contrast media with different iodine concentrations at an identical iodine delivery rate.

PURPOSE: To evaluate the influence of osmolarity of iodinated contrast media and saline flush on the contrast effect in thoracic computed tomography angiography (CTA) at an identical iodine delivery rate (IDR). MATERIALS AND METHODS: Seven beagles were used in a cross-over experiment. The contrast media used were iohexol 350 mgI/ml (IOH350; osmolarity 844 mmol/kg) and iodixanol 320 mgI/ml (IDX320; osmolarity 290 mmol/kg). Each contrast medium was administered to groups with and without saline flush at 40.0 mgI/kg/s for all experiments. Dynamic CT scanning was performed at the ninth thoracic vertebra level. The peak value, area under the curve (AUC), and time to peak (TTP) were calculated from the time attenuation curves of the pulmonary artery and aorta. RESULTS: There was no significant difference between IOH350 and IDX320 with or without saline flush in the peak values for the pulmonary artery and aorta. AUC was significantly higher in groups with saline flush for both contrast media and arteries (p<0.05) with no significant difference between contrast media. TTP was significantly longer in groups with saline flush than without saline flush for both contrast media and arteries (p<0.05), with no significant difference between contrast media. CONCLUSIONS: There were no significant differences in the contrast effects of monomeric IOH350 and dimeric IDX320 in thoracic CTA when used at an identical IDR. Moreover, saline flush prolonged the peak duration at 600 mgI/kg.

Inflammatory reaction in the herniated degenerative disc materials in miniature dachshunds.

Histological and immunohistochemical studies were conducted using herniated intervertebral disc materials obtained surgically from 39 miniature dachshunds. Infiltration of inflammatory cells, such as macrophages, neutrophils, T or B lymphocytes, and multinucleated giant cells, were identified in intervertebral disc materials in 23 cases. Furthermore, proliferations of connective tissue, including neovascularization, were also observed in 17 cases. These results suggest that spontaneous regression of herniated intervertebral disc material could occur in affected dogs.

Effect of an autogenous free fat graft on hemilaminectomy defects in dogs.

The purpose of this study was to evaluate the effect of an autogenous free fat graft (FFG) on hemilaminectomy defects. Hemilaminectomy was performed at three levels in three beagles, and each defect was designated as a control site or a site treated with one of two different sizes of FFG. Subsequently, longitudinal computed tomography scanning and histopathological examination were performed. As a result, no postoperative dural adhesion was recognized at the sites where FFGs were placed regardless of the size of the FFG. Moreover, there was no compression of the spinal cord by the FFG. Therefore, it appears that postoperative dural adhesion was controlled by the FFG and that the possibility of FFG migration into the spinal canal is low in hemilaminectomy. In conclusion, placement of an FFG over the defect was considered useful for preventing complications in hemilaminectomy.

Significance of imaging features of alveolar echinococcosis in studies on nonhuman primates.

In this study, we report the imaging findings in two Japanese monkeys (Macaca fuscata) diagnosed with alveolar echinococcosis. Both monkeys were treated with albendazole for 10 years, without surgery. Radiography, computed tomography, and contrast-enhanced ultrasonography were performed under general anesthesia. This is the first report on contrast-enhanced ultrasonographic imaging for alveolar echinococcosis wherein perflubutane was used as the contrast medium. The findings of the imaging analyses were similar to those reported for alveolar echinococcosis in humans, such as snowflake sign and worm-eaten sign. In addition, the serology correlated well with the imaging data in these two monkeys. Therefore, we propose that the imaging findings of alveolar echinococcosis in nonhuman primates may be used to accumulate data on this condition in human alveolar echinococcosis.

Providing an intelligible explanation to pet owners by using three-dimensional CT images: use of clinical imaging for better informed consent.

The purpose of the study reported here was to discuss the level of intelligibility of three-dimensional computed tomography (3D-CT) images of clinical patients for persons who are lacking veterinary knowledge. 143 undergraduate students were participated in this study. They were entirely unfamiliar with veterinary anatomy. A survey was conducted using the visual analog scale method with a focus on the level of intelligibility of the 3D-CT images compared to that of the two-dimensional CT images or radiographs. As a result, the mean value of the intelligibility of the 3D-CT image was 88.4 +/- 17.8 points out of 100 points. In clinical cases, it is very difficult for pet owners lacking veterinary knowledge to understand the clinical status of their pets even with explanations using illustrations, radiographs, and transverse CT images. It is considered that better informed consent and patient satisfaction will be obtained provided veterinarians make an effort towards intelligible patient education by using 3D-CT images.

Alveolar macrophages are indispensable for controlling influenza viruses in lungs of pigs.

Alveolar macrophages constitutively reside in the respiratory tracts of pigs and humans. An in vivo role of alveolar macrophages in defending against influenza viruses in mice infected with a reassorted influenza virus, 1918 HA/NA:Tx/91, was reported, but there has been no report on an in vivo role of alveolar macrophages in a natural host such as a pig using currently circulating human influenza virus. Here we show that in vivo depletion of alveolar macrophages in pigs by dichloromethylene diphosphonate (MDPCL2) treatment results in 40% mortality when pigs are infected with currently circulating human H1N1 influenza viruses, while none of the infected control pigs died. All infected pigs depleted of alveolar macrophages suffered from more severe respiratory signs than infected control pigs. Induction of tumor necrosis factor alpha in the infected pigs depleted of alveolar macrophages was significantly lower than that in the lungs of infected control pigs, and the induction of interleukin-10, an immunosuppressive cytokine, significantly increased in the lungs of infected pigs depleted of alveolar macrophages compared to infected control pigs. When we measured antibody titers and CD8(+) T lymphocytes expressing gamma interferon (IFN-gamma), lower antibody titers and a lower percentage of CD8(+) T lymphocytes expressing IFN-gamma were detectable in MDPCL2-treated infected pigs than in phosphate-buffered saline- and liposome-treated and infected pigs. Taken together, our findings suggest that alveolar macrophages are essential for controlling H1N1 influenza viruses in pigs.