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Naturwissenschaften ; 109(1): 12, 2022 Jan 07.
Article En | MEDLINE | ID: mdl-34994866

Cities are home to several species of pollinators that play an important role in the reproductive success of wild and cultivated plants that grow in these ecosystems and their surroundings. Pollution is a main driver of pollinator decline. Light and noise pollution are more intense in cities than in any other ecosystem. Although nocturnal pollinators are heavily exposed to these pollutants, their effect on bat pollination is still unknown. Our goal was to assess the effect of light and noise pollution on the main pollination components (pollinator visits, pollen transfer, pollen germination, fruit, and seed set) of the tropical tree, Ceiba pentandra, in a heavily urbanized ecosystem. We measured these components in sites with contrasting intensities of artificial light and anthropogenic noise and statistically assessed the direct and indirect effect of pollutants on pollination components using structural equation modeling. We found that noise and light pollution negatively affected the visits by the bats that pollinate C. pentandra. However, these negative effects did not affect posterior pollination components. In fact, the direct effect of light pollution on reproductive success was positive and greater than the indirect effects via pollinator visits. We suggest that illuminated trees may be able to sustain a large quantity of fruits and seeds because they produce more photosynthates due to greater light radiation and delayed leaf abscission. We conclude that, despite the negative effect of light and noise on pollinator visits, these pollutants did not significantly impact the reproductive success of C. pentandra.

Chiroptera , Trees , Animals , Ecosystem , Flowers , Pollination , Reproduction
BMC Genomics ; 23(1): 33, 2022 Jan 07.
Article En | MEDLINE | ID: mdl-34996349

BACKGROUND: Long non-coding RNAs (lncRNAs) play a critical role in the pathogenesis of hypoxic pulmonary hypertension (HPH). The role of N7-methylguanosine (m7G) modification in lncRNAs has received increased attentions in recent years. However, the m7G-methylation of lncRNA in HPH has yet to be determined. We have therefore performed a transcriptome-wide analysis of m7G lncRNAs in HPH. RESULTS: Differentially-expressed m7Gs were detected in HPH, and m7G lncRNAs were significantly upregulated compared with non-m7G lncRNAs in HPH. Importantly, this was the first time that the upregulated m7G lncXR_591973 and m7G lncXR_592398 were identified in HPH. CONCLUSION: This study provides the first m7G transcriptome-wide analysis of HPH. Importantly, two HPH-associated m7G lncRNAs were identified, although their clinical significance requires further validation.

Hypertension, Pulmonary , RNA, Long Noncoding , Animals , Guanosine/analogs & derivatives , Hypertension, Pulmonary/genetics , Hypoxia/genetics , RNA, Long Noncoding/genetics , Rats
BMC Genomics ; 23(1): 36, 2022 Jan 07.
Article En | MEDLINE | ID: mdl-34996352

BACKGROUND: Bioassessment and biomonitoring of meat products are aimed at identifying and quantifying adulterants and contaminants, such as meat from unexpected sources and microbes. Several methods for determining the biological composition of mixed samples have been used, including metabarcoding, metagenomics and mitochondrial metagenomics. In this study, we aimed to develop a method based on next-generation DNA sequencing to estimate samples that might contain meat from 15 mammalian and avian species that are commonly related to meat bioassessment and biomonitoring. RESULTS: In this project, we found the meat composition from 15 species could not be identified with the metabarcoding approach because of the lack of universal primers or insufficient discrimination power. Consequently, we developed and evaluated a meat mitochondrial metagenomics (3MG) method. The 3MG method has four steps: (1) extraction of sequencing reads from mitochondrial genomes (mitogenomes); (2) assembly of mitogenomes; (3) mapping of mitochondrial reads to the assembled mitogenomes; and (4) biomass estimation based on the number of uniquely mapped reads. The method was implemented in a python script called 3MG. The analysis of simulated datasets showed that the method can determine contaminant composition at a proportion of 2% and the relative error was < 5%. To evaluate the performance of 3MG, we constructed and analysed mixed samples derived from 15 animal species in equal mass. Then, we constructed and analysed mixed samples derived from two animal species (pork and chicken) in different ratios. DNAs were extracted and used in constructing 21 libraries for next-generation sequencing. The analysis of the 15 species mix with the method showed the successful identification of 12 of the 15 (80%) animal species tested. The analysis of the mixed samples of the two species revealed correlation coefficients of 0.98 for pork and 0.98 for chicken between the number of uniquely mapped reads and the mass proportion. CONCLUSION: To the best of our knowledge, this study is the first to demonstrate the potential of the non-targeted 3MG method as a tool for accurately estimating biomass in meat mix samples. The method has potential broad applications in meat product safety.

Genome, Mitochondrial , Metagenomics , Animals , Mammals , Meat , Sequence Analysis, DNA
BMC Surg ; 22(1): 6, 2022 Jan 08.
Article En | MEDLINE | ID: mdl-34996410

BACKGROUND: Ingestion of fish bones leading to gastric perforation and inducing abscess formation in the caudate lobe of the liver is very rare. CASE PRESENTATION: A 67-year-old man presented to our hospital with a 2-day history of subxiphoid pain. There were no specific symptoms other than pain. Laboratory tests showed only an increase in the number and percentage of neutrophils. Contrast-enhanced Computerized tomography (CT) of the abdomen showed two linear dense opacities in the gastric cardia, one of which penetrated the stomach and was adjacent to the caudate lobe of the liver, with inflammatory changes in the caudate lobe. We finally diagnosed his condition as a caudate lobe abscess secondary to intestinal perforation caused by a fishbone based on the history and imaging findings. The patient underwent 3D laparoscopic partial caudate lobectomy, incision and drainage of the liver abscess, and fishbone removal. The procedure was successful and we removed the fishbone from the liver. The patient was discharged on the 9th postoperative day without other complications. CONCLUSIONS: Liver abscess caused by foreign bodies requires multidisciplinary treatment. Especially when located in the caudate lobe, we must detect and remove the cause of the abscess as early as possible. Foreign bodies that perforate the gastrointestinal tract can penetrate to the liver and cause abscess formation, as in this case. When exploring the etiology of liver abscesses, we should investigate the general condition, including the whole gastrointestinal tract.

Foreign Bodies , Foreign-Body Migration , Laparoscopy , Liver Abscess , Aged , Animals , Foreign Bodies/complications , Foreign Bodies/diagnostic imaging , Foreign Bodies/surgery , Foreign-Body Migration/complications , Foreign-Body Migration/diagnostic imaging , Foreign-Body Migration/surgery , Humans , Liver Abscess/diagnostic imaging , Liver Abscess/etiology , Liver Abscess/surgery , Male
BMJ Case Rep ; 15(1)2022 Jan 07.
Article En | MEDLINE | ID: mdl-34996770

Postoperative fevers are common in hospitalised patients and warrant workup beyond the early post-op period. A 50-year-old man was admitted after sustaining a tibial plateau fracture. Fevers began 3 days after external fixation and persisted through a second surgery despite initial negative workup. Careful review of medications revealed enoxaparin as the instigating agent of a febrile drug reaction, and the fevers resolved after discontinuing the drug. On further questioning, it was discovered the patient had an allergy to pork, from which the main components of enoxaparin are typically derived. To our knowledge, this is the first reported enoxaparin-induced fever in the setting of a pork allergy. Enoxaparin-induced fevers should be considered in patients with unexplained post-op fever. Our case demonstrates the importance of analysing newly administered medications. Simple detailed history may significantly reduce patient morbidity and help to broaden differentials during investigation.

Fever of Unknown Origin , Hypersensitivity , Pork Meat , Red Meat , Animals , Anticoagulants/adverse effects , Enoxaparin/adverse effects , Humans , Male , Middle Aged , Postoperative Complications/chemically induced , Swine
Int J Nanomedicine ; 17: 91-104, 2022.
Article En | MEDLINE | ID: mdl-35027828

Purpose: Traumatic spinal cord injury (TSCI) induces a powerful inflammatory response that can significantly exacerbate the extent and severity of neural damage (termed as "secondary injury"). Thus, the suppression of inflammation is crucial for reducing neurological dysfunction following TSCI. However, the conventional anti-inflammatory drugs show limited efficacy because of poor penetration and release kinetics at the injury site. This study describes the design, synthesis, release kinetics, biosafety, and preclinical efficacy of minocycline (MC)-loaded poly(α-lipoic acid)-methylprednisolone (PαLA-MP) prodrug nanoparticles (NPs) for the combined anti-inflammatory treatment of TSCI. Methods: NPs were produced by conjugating MP to PαLA and then loading MC. The NP structure was confirmed through 1H nuclear magnetic resonance (1H NMR), Fourier transform infrared spectroscopy, ultraviolet-visible absorption spectroscopy, gel permeation chromatography, dynamic light scattering, and transmission electron microscopy. Drug-loading content and efficacy were measured using high-performance liquid chromatography (HPLC) or 1H NMR and release kinetics through HPLC. Biosafety was examined using the MTT assay, cell penetration efficiency using confocal microscopy, and flow cytometry using Cyanine5 (Cy5)-labeled MC-PαLA-MP NPs, effects on injury-induced pro-inflammatory cytokine release using enzyme-linked immunosorbent assays and immunofluorescence, and treatment efficacy by measuring motor recovery in a rat model of TSCI. Results: The MC-PαLA-MP NPs exhibited high biocompatibility and released 81% MC and 54% MP within 24 h under TSCI-like conditions, effectively reducing 40% of pro-inflammatory cytokine release both in cultures and injured rat spinal cord tissues. Systemic injection increased the Basso, Beattie, Bresnahan score of TSCI rats from 2.33 ± 0.52 to 8.83 ± 1.83 in 8 weeks, providing effective neuroprotection and enhanced exercise recovery in the TSCI rats. Conclusion: The MC-PαLA-MP NPs can mitigate secondary inflammation and preserve motor function following experimental TSCI, which suggests their potential for clinical application.

Nanoparticles , Prodrugs , Spinal Cord Injuries , Thioctic Acid , Animals , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/therapeutic use , Methylprednisolone , Minocycline , Prodrugs/therapeutic use , Rats , Recovery of Function , Spinal Cord , Spinal Cord Injuries/drug therapy
Trop Anim Health Prod ; 54(1): 54, 2022 Jan 14.
Article En | MEDLINE | ID: mdl-35028725

The aim of this study was to evaluate the effects of dietary vitamin C supplementation on blood parameters of pre-parturient (PP) dairy cows and growth performance and immune system of their newborn calves. Forty PP cows (at approximately 21 days before calving and an average weight 791 ± 50 kg) were allocated into two experimental treatments: (1) basal diet without vitamin C supplementation (CO) and (2) basal diet with 20 g of vitamin C supplementation from 21 days before calving to parturition (VC). After parturition, the experiment continued by grouping the calves into four dietary treatments with 8 calves in each treatment. The experimental treatments were (1) control calves with no vitamin C supplementation and from cows that received no vitamin C supplement (CON), (2) calves supplemented with 600 mg of vitamin C per day and from cows that received no vitamin C supplement (CVC), (3) calves supplemented with no vitamin C and from cows that received 20 g of vitamin C per day (MVC), and (4) calves supplemented with 600 mg of vitamin C per day and from cows that received 20 g of vitamin C per day (CMVC). Serum concentrations of glucose, HDL and LDL, cholesterol, triglycerides, total protein, and albumin of cows were not affected by vitamin C supplementation during pre-parturient period. However, cows that received VC diet had lower (P < 0.05) malondialdehyde (MDA) and aspartate aminotransferase (AST) concentrations, higher total antioxidant capacity (TAC), and vitamin C concentration in their blood compared to CO cows. Vitamin C supplementation had no effect on plasma iron, copper, and zinc concentrations of PP cows. Similarly, vitamin C supplementation had no effect on total feed intake and feed conversion ratio (FCR) of suckling calves. However, calves in the CMVC group had higher (P < 0.05) overall daily weight gain compared to the other groups. Calves in the CVC and CMVC groups had lower (P < 0.05) blood MDA concentration on days 7 and 21. The highest (P < 0.05) blood TAC level was recorded in CMVC calves. Control group calves had lower (P < 0.05) blood superoxide dismutase activity compared to the other calves. Blood levels of alanine aminotransferase on days 7 and 21 and aspartate aminotransferase on day 7 were higher (P < 0.05) for calves in the CON and MVC groups. Based on the results, vitamin C supplementation had positive health effects on the oxidative parameters of PP dairy cows and also improved the performance and health status of the calves.

Copper , Zinc , Animal Feed/analysis , Animals , Ascorbic Acid , Cattle , Diet/veterinary , Dietary Supplements , Female , Iron
AAPS PharmSciTech ; 23(1): 54, 2022 Jan 13.
Article En | MEDLINE | ID: mdl-35028813

The purpose of our study was using a computational simulation to develop a long-acting patch of rivastigmine (RVS). A range of patch formulations were screened including pressure sensitive adhesive (PSA), pharmaceutical excipients, and controlled release membranes using transfer simulation based on a mathematical model. Diffusion dynamics parameters for simulated operations were acquired through in vitro release tests (IVRT) and in vitro skin permeation tests (IVPT). The mechanism of controlled release was studied by FTIR (Fourier transform infrared), DSC (differential scanning calorimeter) and molecular docking. Results of a rat in vitro permeation profile showed excellent correlation with the in vivo deconvolution profile (R2=0.998). Experiments testified to transfer of RVS at a relatively uniform speed with high skin permeation (2531.2±142.46 µg/cm2) in 72 h. Pharmacokinetic data obtained in vivo also confirmed stable plasma concentrations over 72 h for the optimized patch, and significant prolongation of both Tmax (11.20±1.79 h) and MRT0-t (33.91±5.33 h). Cmax was controlled with AUC0-t (267.34±24.46 h ng/ml), which was closely comparable to parameters of a commercial Exelon® Patch. The successful development of a long-acting patch of RVS thus underscores the potential of computer aided design in a context of promnesic transdermal delivery. Graphical abstract.

Skin Absorption , Transdermal Patch , Administration, Cutaneous , Animals , Molecular Docking Simulation , Rats , Rats, Wistar , Rivastigmine , Skin/metabolism
Chin J Dent Res ; 24(4): 251-256, 2022 Jan 14.
Article En | MEDLINE | ID: mdl-35029096

OBJECTIVE: To assess bone volume stability and identify critical bone graft thickness for guided bone regeneration (GBR) simultaneous to implant placement in the maxillary anterior region. METHODS: Eighteen patients were included in the study and received placement of one maxillary anterior implant combined with GBR using deproteinised bovine bone mineral (DBBM) and resorbable collagen membrane. The multilevel buccal bone thickness (BT) was measured by CBCT 1 to 2 weeks and 5 to 8 months post-implantation (at implant uncovering surgery). RESULTS: Significant buccal alveolar crest collapse (ACCb 1.20 to 1.70 mm) occurred during the mean healing period of 5.3 months (P = 0.000). A greater percentage of ACCb occurred at the coronal aspect of the implant, with 59.24% ± 19.22% at the implant platform and 34.10% ± 30.50% 6.0 mm below the implant platform, respectively. Linear regression analysis demonstrated that if BT was 1.8 to 2.0 mm at the implant platform at uncovering surgery, then it was estimated to have been 4.1 to 4.5 mm immediately post-implantation. CONCLUSION: ACCb after maxillary anterior implant placement with simultaneous GBR occurred more coronally than apically. Excessive alveolar ridge augmentation, up to 4.0 mm of hard tissue buccal to the implant platform, should be achieved at the time of surgery to compensate for this potential resorption.

Alveolar Ridge Augmentation , Dental Implants , Animals , Bone Regeneration , Cattle , Dental Implantation, Endosseous , Humans , Maxilla/diagnostic imaging , Maxilla/surgery
In Vivo ; 36(1): 86-93, 2022.
Article En | MEDLINE | ID: mdl-34972703

BACKGROUND/AIM: Brown recluse spider bite releases hemolytic and cytotoxic phospholipase D to the wound that may cause necrosis or even death. We examined diethyl azelate (DEA), a plasma membrane fluidizer with a broad range of immunomodulatory activities, as a potential treatment for the brown recluse spider bite. MATERIALS AND METHODS: Topical DEA was used in emergency to treat brown recluse spider bites in a human subject. We subsequently evaluated the effects of DEA on hemolysis induced by the brown recluse spider venom, recluse recombinant phospholipase D (rPLD), and venoms from honey bee and moccasin snake, and on phospholipase A2 activity in the bee and snake venoms and in human urine. RESULTS: Topical DEA resolved the consequences of human brown recluse spider envenomation in two weeks. In vitro, DEA inhibited hemolysis caused by the brown recluse spider venom and rPLD and suppressed phospholipase A2 activity in a dose-dependent manner. CONCLUSION: DEA is a promising novel therapy for the brown recluse spider bite and perhaps even unrelated envenomations involving PLDs.

Brown Recluse Spider , Spider Bites , Animals , Dicarboxylic Acids , Hemolysis , Necrosis , Spider Bites/drug therapy
In Vivo ; 36(1): 170-179, 2022.
Article En | MEDLINE | ID: mdl-34972712

BACKGROUND/AIM: Cancer cell inoculation is routinely used to evaluate novel therapeutic approaches in vivo. However, without reporter genes enabling deep tissue imaging, study of early tumor progression and therapeutic responses is often limited. We describe the establishment and characterization of two canine cancer cell lines stably expressing red fluorescence proteins as tools for later in vivo imaging. MATERIALS AND METHODS: Two red fluorescence cell lines were generated by plasmid transfection. Fluorescence protein expression was confirmed by flow cytometry and microscopy. Deep tissue imaging was demonstrated in mice using a NightOWL LB 983. Gene expression changes after transfection were analyzed by RNAseq. RESULTS: Both cell lines were detectable in vivo by subcutaneous injection of 1×106 cells. RNAseq revealed up to 2005 transfection-induced differentially expressed genes but no significant changes in cellular key pathways. CONCLUSION: The fluorescent cell lines provide a solid basis for future in vivo studies on canine cancer.

Adenocarcinoma , Carcinoma, Transitional Cell , Prostatic Neoplasms , Adenocarcinoma/genetics , Animals , Cell Line, Tumor , Dogs , Humans , Male , Mice , Prostate , Prostatic Neoplasms/genetics , Transfection
Trop Anim Health Prod ; 54(1): 34, 2022 Jan 03.
Article En | MEDLINE | ID: mdl-34978613

The objective of this study was to contribute to genetic improvement for carcass quality in Nellore cattle by estimating genetic parameters for carcass characteristics measured by traditional and novel ultrasound adjusted for body weight. Records of 19,910 Nellore breed animals born between 2001 and 2015 as part of a genetic improvement program were evaluated for this purpose. The (co)variance components and genetic parameters for yearling weight (YW), slaughter conformation at yearling age (SCY), ribeye area (REA), REA adjusted for 100 kg of body weight (REA100), REA adjusted for 450 kg of body weight (REA450), subcutaneous fat thickness over the loin (SFTL) and the rump (SFTR), intramuscular fat (IF), and height × width ratio of the sirloin steak (RATIO) were estimated using the restricted maximum likelihood (REML) method and the three-trait animal model. The estimated heritability values were 0.39 (YW), 0.33 (SCY), 0.31 (REA), 0.42 (REA100), 0.38 (REA450), 0.31 (RATIO), 0.17 (SFTL), 0.27 (SFTR), and 0.41 (MAR). The genetic correlations between REA and YW (0.43) and SCY and YW (0.78), respectively, suggested that selecting for REA or SCY could result in an increase in body weight. Because of its moderate and negative correlation with YW, REA100 was ineffective as a muscling indicator (- 0.38). The indicators REA450 and RATIO had a - 0.12 and 0.11 low correlation with YW, respectively. Thus, it is concluded that REA450 and RATIO were the best indicators of muscling, with heritability of medium magnitude, making the selection viable, as well as having a low correlation with body weight.

Models, Genetic , Muscles , Animals , Body Weight , Cattle/genetics , Phenotype , Ultrasonography/veterinary
Arch Microbiol ; 204(1): 108, 2022 Jan 03.
Article En | MEDLINE | ID: mdl-34978619

Fall armyworm (FAW), Spodoptera frugiperda (Smith, 1797), is a polyphagous, voracious, and economically important agricultural pest. Biological control of FAW is a strategy that must be further explored. This study evaluated six baculovirus strains isolated from infected FAW larvae from Mexico, Argentina, Honduras, and the United States. Five alphabaculoviruses (SfNPV-An2, SfNPV-Arg, SfNPV-Fx, SfNPV-Ho, and SfNPV-Sin) and one betabaculovirus (SfGV-RV) were tested against FAW larvae, showing a wide diversity of virulence levels among strains when their estimated LC50s were compared, being SfNPV-Arg, SfNPV-Ho and SfNPV-Fx more virulent than SfNPV-An2, SfNPV-Sin, and SfGV-RV. To determine any virulence difference in vitro studies of these isolates, Sf9 cell cultures were used. Interestingly, only ODVs from four of the test SfNPV strains showed infectivity on Sf9 cell cultures, and some differences in virulence were observed. Genomic restriction analyses and partial sequences of lef-8, lef-9, and polh/granulin genes showed little variability among alphabaculoviruses, both, among them and with previously reported sequences. However, sequences from SfGV-RV were closer to previously reported sequences from the SfGV-VG008 strain than the SfGV-Arg and SfGV-VG014 strains. The great difference in the in vivo virulence was not correlated with great similarity among the isolates. The characterization of these six baculovirus isolates offers the basis for exploring their potential as biological control agents against S. frugiperda, as well the initial studies on their specific infection mechanisms, evolution, and ecology.

Baculoviridae , Moths , Animals , Baculoviridae/genetics , Larva , Spodoptera , Virulence , Zea mays
Gen Dent ; 70(1): 30-33, 2022.
Article En | MEDLINE | ID: mdl-34978987

The purpose of this in vitro study was to compare the 3- and 90-day bond strengths of 3 cements used for luting metal-ceramic crowns. Zinc phosphate cement (ZPC; SS White), resin-modified glass ionomer cement (RMGIC; Fuji Plus), and self-adhesive resin cement (SARC; RelyX U200) were assessed in 2 different treatment conditions (with and without microsandblasting of the alloy) and at 2 experimental times (3 days [E1] and 90 days [E2] after cementation). The buccal surfaces of 84 bovine teeth were ground until the dentin was exposed, and 84 nickel-chromium alloy plates cast from a resin model were cemented to the dentin surfaces with 1 of the 3 cements (n = 28). In half of the specimens of each group (n = 14), the bonding surfaces of the nickel-chromium plates received 6 seconds of microsandblasting with 45-µm aluminum oxide particles prior to cementation. The compressive shear bond strengths of the specimens were evaluated in a universal testing machine at E1 and E2 (n = 7). The SARC group showed the greatest bond strength, followed by the RMGIC group, while the bond strength of the ZPC group was significantly lower (P < 0.01). For the RMGIC specimens subjected to microsandblasting, there was a statistically significant difference between the mean bond strengths at E1 and E2 (P = 0.040). All of the other cement and treatment groups showed statistically similar adhesion results at E1 and E2 (P > 0.05). The complementary test by Sidák revealed that the cements Fuji Plus and RelyX U200 showed higher values at E2 and were statistically similar to each other (P > 0.05). Although RMGIC specimens showed a lower initial bond strength than SARC specimens, the fact that the microsandblasted RMGIC subgroup was the only one that demonstrated a significant increase in bond strength with aging suggests that RMGIC can be a material of first choice because it also costs less than SARC.

Dental Bonding , Animals , Cattle , Chromium Alloys , Dental Cements/therapeutic use , Dental Stress Analysis , Glass Ionomer Cements , Humans , Materials Testing , Resin Cements , Surface Properties
Gen Dent ; 70(1): 46-50, 2022.
Article En | MEDLINE | ID: mdl-34978990

This study evaluated the influence of different combinations of 3 final endodontic irrigation techniques (positive-pressure irrigation [PPI], passive ultrasonic irrigation [PUI], and continuous ultrasonic irrigation [CUI]) and 2 irrigants (5.25% sodium hypochlorite [NaOCl] and 2% chlorhexidine [CHX]) on the push-out resistance of glass fiber posts. Ninety bovine roots were prepared and divided into 6 groups (n = 15): PPIN, PPI with 5.25% NaOCl; PUIN, PUI with 5.25% NaOCl; CUIN, CUI with 5.25% NaOCl; PPIC, PPI with 2% CHX; PUIC, PUI with 2% CHX; and CUIC, CUI with 2% CHX. In the PPI groups, a 30-gauge needle was used to deliver the irrigating solution. In the PUI groups, a 30-gauge needle was used to deliver the irrigating solution, which was activated for 10 seconds by means of a stainless steel file mounted on an ultrasonic unit. In the CUI groups, delivery and continuous agitation of the irrigating solution for 10 seconds were performed using a stainless steel file mounted on the ultrasonic unit. After canal preparation was completed, glass fiber posts were cemented with a dual-cure resin cement. The roots were sectioned into 1-mm-thick slices (coronal, middle, and apical thirds). After 7 days' storage in distilled water, the slices were submitted to a push-out test. The mean bond strength values were compared using 2-way analysis of variance and Tukey post hoc test (P < 0.05). The mean (SD) overall bond strength values of the specimens were as follows: PPIN, 4.46 (1.43) MPa; PUIN, 6.13 (2.35) MPa; CUIN, 7.41 (1.83) MPa; PPIC, 3.59 (1.70) MPa; PUIC, 5.49 (1.41) MPa; and CUIC, 6.0 (1.63) MPa. Irrespective of the irrigating solution, continuous ultrasonic irrigation achieved significantly greater push-out bond strengths for glass fiber posts, mainly at the apical third of the root specimens (P < 0.05).

Dental Bonding , Post and Core Technique , Animals , Cattle , Dental Pulp Cavity , Dentin , Glass , Humans , Materials Testing , Resin Cements , Root Canal Irrigants , Root Canal Preparation
J Headache Pain ; 23(1): 1, 2022 Jan 03.
Article En | MEDLINE | ID: mdl-34979902

BACKGROUND: Chronic migraine places a disabling burden on patients, which is extensively modeled by the nitroglycerin (NTG)-treated animal model. Although the NF-κB pathway is involved in an increase in CGRP levels and activation of the trigeminal system in the NTG model, the relationship between NTG and neuroinflammation remains unclear. This study aimed to optimize a chronic NTG rat model with hyperalgesia and the ethological capacity for estimating migraine therapies and to further explore the underlying mechanism of NTG-induced migraine. METHODS: Rats were administered different doses of NTG s.c. daily or every 2 d; 30 min and 2 h later, the mechanical threshold was tested. After 9 d, the rats were injected with EB or Cy5.5 for the permeability assay. The other animals were sacrificed, and then, brainstem and caudal trigeminal ganglion were removed to test CGRP, c-Fos and NOS activity; Cytokines levels in the tissue and serum were measured by ELISA; and NF-κB pathway and blood-brain barrier (BBB)-related indicators were analyzed using western blotting. Immunohistochemistry was performed to observe microglial polarization and IL-17A+ T cell migration in the medulla oblongata. RESULTS: NTG (10 mg/kg, s.c., every 2 d for a total of 5 injections) was the optimal condition, resulting in progressive hyperalgesia and migraine behavior. TNC neuroinflammation with increases in cytokines, CGRP and c-Fos and activation of the NF-κB pathway was observed, and these changes were alleviated by ibuprofen. Furthermore, NTG administration increased BBB permeability by altering the levels functional proteins (RAGE, LRP1, AQP4 and MFSD2A) and structural proteins (ZO-1, Occludin and VE-cadherin-2) to increase peripheral IL-17A permeation into the medulla oblongata, activating microglia and neuroinflammation, and eventually causing hyperalgesia and migraine attack. CONCLUSIONS: This study confirmed that NTG (10 mg/kg, s.c., every 2 d for a total of 5 injections) was the optimal condition to provoke migraine, resulting in mechanical hyperalgesia and observable migraine-like behavior. Furthermore, IL-17A crossed the blood-brain barrier into the medulla oblongata, triggering TNC activation through microglia-mediated neuroinflammation. This process was a novel mechanism in NTG-induced chronic migraine, suggesting that IL-17A might be a novel target in the treatment of migraine.

Migraine Disorders , Nitroglycerin , Animals , Blood-Brain Barrier , Disease Models, Animal , Humans , Interleukin-17 , Migraine Disorders/chemically induced , Nitroglycerin/toxicity , Rats
J Extracell Vesicles ; 11(1): e12179, 2022 01.
Article En | MEDLINE | ID: mdl-34982509

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) entry is mediated by the interaction of the viral spike (S) protein with angiotensin-converting enzyme 2 (ACE2) on the host cell surface. Although a clinical trial testing soluble ACE2 (sACE2) for COVID-19 is currently ongoing, our understanding of the delivery of sACE2 via small extracellular vesicles (sEVs) is still rudimentary. With excellent biocompatibility allowing for the effective delivery of molecular cargos, sEVs are broadly studied as nanoscale protein carriers. In order to exploit the potential of sEVs, we design truncated CD9 scaffolds to display sACE2 on the sEV surface as a decoy receptor for the S protein of SARS-CoV-2. Moreover, to enhance the sACE2-S binding interaction, we employ sACE2 variants. sACE2-loaded sEVs exhibit typical sEVs characteristics and bind to the S protein. Furthermore, engineered sEVs inhibit the entry of wild-type (WT), the globally dominant D614G variant, Beta (K417N-E484K-N501Y) variant, and Delta (L452R-T478K-D614G) variant SARS-CoV-2 pseudovirus, and protect against authentic SARS-CoV-2 and Delta variant infection. Of note, sACE2 variants harbouring sEVs show superior antiviral efficacy than WT sACE2 loaded sEVs. Therapeutic efficacy of the engineered sEVs against SARS-CoV-2 challenge was confirmed using K18-hACE2 mice. The current findings provide opportunities for the development of new sEVs-based antiviral therapeutics.

Angiotensin-Converting Enzyme 2/immunology , COVID-19/immunology , Extracellular Vesicles/immunology , Spike Glycoprotein, Coronavirus/immunology , Animals , Female , HEK293 Cells , Humans , Mice , Protein Binding , Protein Interaction Domains and Motifs
BMC Complement Med Ther ; 22(1): 7, 2022 Jan 04.
Article En | MEDLINE | ID: mdl-34983472

BACKGROUND: Patrinia scabra Bunge is a well-known herbal medicine for its favorable treatment on inflammatory diseases owing to its effective ingredients, in which iridoid glycoside plays an extremely significant role. This article aimed to improve the content of total iridoid glycosides in crude extract through a series optimization of extraction procedure. Moreover, considering that both pain and inflammation are two correlated responses triggered in response to injury, irritants or pathogen, the article investigated the anti-inflammatory and analgesic activities of P. scabra to screen out the active fraction. METHOD: P. scabra was extracted by ultrasonic-microwave synergistic extraction (UMSE) to obtain total iridoid glycosides (PSI), during which a series of conditions were investigated based on single-factor experiments. The extraction process was further optimized by a reliable statistical method of response surface methodology (RSM). The elution fractions of P. scabra extract were prepared by macroporous resin column chromatography. Through the various animal experiment including acetic acid-induced writhing test, formalin induced licking and flinching, carrageenan-induced mice paw oedema test and xylene-induced ear edema in mice, the active fractions with favorable analgesic and anti-inflammatory effect were reasonably screen out. RESULTS: The content of PSI could reach up to 81.42 ± 0.31 mg/g under the optimum conditions as follows: ethanol concentration of 52%, material-to-liquid ratio of 1:18 g/mL, microwave power at 610 W and extraction time of 45 min. After gradient elution by the macroporous resin, the content of PSI increased significantly. Compared with other concentrations of elution liquid, the content of PSI in 30 and 50% ethanol eluate was increased to reach 497.65 and 506.90 mg/g, respectively. Owing to the pharmacology experiment, it was reasonably revealed that 30 and 50% ethanol elution fractions of P. scabra could relieve pain centrally and peripherally, exhibiting good analgesic and anti-inflammatory activities. CONCLUSION: Patrinia scabra possessed rich iridoids and exhibited significant analgesic and anti-inflammatory activities.

Anti-Inflammatory Agents/pharmacokinetics , Iridoid Glycosides/pharmacology , Iridoids/pharmacology , Microwaves , Patrinia/metabolism , Ultrasonics , Analgesics/pharmacology , Animals , Anti-Inflammatory Agents/pharmacology , Inflammation/drug therapy , Iridoids/therapeutic use , Mice , Pain/drug therapy , Phytotherapy , Plants, Medicinal/metabolism
BMC Complement Med Ther ; 22(1): 5, 2022 Jan 04.
Article En | MEDLINE | ID: mdl-34983484

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.

Gastrointestinal Microbiome/drug effects , Lipids/blood , Ovariectomy , Plant Extracts/metabolism , Trifolium/metabolism , Animals , Rats , Rats, Sprague-Dawley
BMC Complement Med Ther ; 22(1): 4, 2022 Jan 04.
Article En | MEDLINE | ID: mdl-34983490

BACKGROUND: Though Lippia javanica (Burm.f.) Spreng antioxidant activity has been demonstrated, its effect in protecting the brain from lead (Pb)-induced oxidative damage is unknown. This study investigated the effect of L. javanica against Pb-induced oxidative stress, inflammation, apoptosis and acetylcholinesterase activity in rat's brain. METHODS: L. javanica herbal tea infusion was prepared, its phytochemical constituent was revealed by liquid chromatography-Mass spectrometer (LC-MS) and was administered simultaneously with Pb. Four groups of male Wistar rats (n = 5/group) were used: control received distilled water; Pb-acetate group received 50 mg Pb/ Kg bodyweight (bw), treatment group received 50 mg Pb/ Kg Pb-acetate + 5 ml/kg bw L. javanica and L. javanica group received 5 ml/Kg bw of L. javanica tea infusion only. After 6 weeks of treatment, oxidative status, acetylcholinesterase activity, inflammation and apoptosis was assessed in brain tissue which was also histologically examined. RESULTS: Mean brain and heart weight was reduced (p < 0.05) while liver and spleen weights were increased (p < 0.05) in Pb exposed animals but were prevented by L. juvanica treatment. Treatment with L. javanica increased (p < 0.05) overall brain antioxidant status (glutathione and superoxide dismutase activities) and reduced lipid peroxidation (p < 0.05) compared to the Pb exposed animals. Pro-inflammatory cytokine tumour necrotic factor-alpha, pro-apoptosis Bax protein and anticholinesterase activity were reduced (p < 0.05) in Pb-L. javanica treated animals compared to the Pb exposed group. Histological examination confirmed neuroprotective effects of L. javanica as evidenced by reduced apoptosis/necrosis and inflammation-induced vacuolization and oedema in the hippocampus. The L. javanica treatment alone had no detrimental effects to the rats. LC-MS analysis revealed L. javanica to be rich in phenolics. CONCLUSIONS: This study demonstrated that L. javanica, rich in phenolics was effective in reducing Pb-induced brain oxidative stress, inflammation, apoptosis, acetylcholinesterase activity and neuronal damage.

Brain/metabolism , Lead/adverse effects , Lippia/metabolism , Oxidation-Reduction , Oxidative Stress/drug effects , Teas, Herbal , Animals , Plant Extracts/pharmacology , Rats , Rats, Wistar