Discovery of Anti-Coronavirus Cinnamoyl Triterpenoids Isolated from Hippophae rhamnoides during a Screening of Halophytes from the North Sea and Channel Coasts in Northern France.

The limited availability of antiviral therapy for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has spurred the search for novel antiviral drugs. Here, we investigated the potential antiviral properties of plants adapted to high-salt environments collected in the north of France. Twenty-five crude methanolic extracts obtained from twenty-two plant species were evaluated for their cytotoxicity and antiviral effectiveness against coronaviruses HCoV-229E and SARS-CoV-2. Then, a bioguided fractionation approach was employed. The most active crude methanolic extracts were partitioned into three different sub-extracts. Notably, the dichloromethane sub-extract of the whole plant Hippophae rhamnoides L. demonstrated the highest antiviral activity against both viruses. Its chemical composition was evaluated by ultra-high performance liquid chromatography (UHPLC) coupled with mass spectrometry (MS) and then it was fractionated by centrifugal partition chromatography (CPC). Six cinnamoyl triterpenoid compounds were isolated from the three most active fractions by preparative high-performance liquid chromatography (HPLC) and identified by high resolution MS (HR-MS) and mono- and bi-dimensional nuclear magnetic resonance (NMR). Specifically, these compounds were identified as 2-O-trans-p-coumaroyl-maslinic acid, 3ß-hydroxy-2α-trans-p-coumaryloxy-urs-12-en-28-oic acid, 3ß-hydroxy-2α-cis-p-coumaryloxy-urs-12-en-28-oic acid, 3-O-trans-caffeoyl oleanolic acid, a mixture of 3-O-trans-caffeoyl oleanolic acid/3-O-cis-caffeoyl oleanolic acid (70/30), and 3-O-trans-p-coumaroyl oleanolic acid. Infection tests demonstrated a dose-dependent inhibition of these triterpenes against HCoV-229E and SARS-CoV-2. Notably, cinnamoyl oleanolic acids displayed activity against both SARS-CoV-2 and HCoV-229E. Our findings suggest that Hippophae rhamnoides could represent a source of potential antiviral agents against coronaviruses.

Luteolin Isolated from Juncus acutus L., a Potential Remedy for Human Coronavirus 229E.

The COVID-19 pandemic, caused by SARS-CoV-2, addressed the lack of specific antiviral drugs against coronaviruses. In this study, bioguided fractionation performed on both ethyl acetate and aqueous sub-extracts of Juncus acutus stems led to identifying luteolin as a highly active antiviral molecule against human coronavirus HCoV-229E. The apolar sub-extract (CH2Cl2) containing phenanthrene derivatives did not show antiviral activity against this coronavirus. Infection tests on Huh-7 cells, expressing or not the cellular protease TMPRSS2, using luciferase reporter virus HCoV-229E-Luc showed that luteolin exhibited a dose-dependent inhibition of infection. Respective IC50 values of 1.77 µM and 1.95 µM were determined. Under its glycosylated form (luteolin-7-O-glucoside), luteolin was inactive against HCoV-229E. Time of addition assay showed that utmost anti-HCoV-229E activity of luteolin was achieved when added at the post-inoculation step, indicating that luteolin acts as an inhibitor of the replication step of HCoV-229E. Unfortunately, no obvious antiviral activity for luteolin was found against SARS-CoV-2 and MERS-CoV in this study. In conclusion, luteolin isolated from Juncus acutus is a new inhibitor of alphacoronavirus HCoV-229E.

Balloon catheter occlusion of the maxillary, internal, and external carotid arteries in standing horses.

OBJECTIVE: To assess the feasibility of balloon catheter occlusion of the internal carotid artery (ICA), external carotid artery (ECA), and maxillary artery (MA) in standing horses. STUDY DESIGN: Experimental and clinical cases series. ANIMALS: Eight healthy horses (phase 1) and 11 clinical cases (phase 2). METHODS: Occlusions were performed on standing horses under sedation and local anesthesia. In phase 1, four horses underwent bilateral ICA balloon catheter occlusion, and four horses underwent balloon catheter occlusion of the ECA and MA. In phase 2, horses were treated by occlusion of ICA (n = 7), ECA (n = 2), or ECA and ICA (n = 2). RESULTS: Internal carotid artery occlusion was successful in seven of eight and seven of nine arteries in phases 1 and 2, respectively. The procedures lasted 53 and 50 minutes, respectively, and catheters were inserted over 13 ± 0.7 cm (mean ± SD). External carotid artery occlusion was successful in seven of seven and four of four arteries in phases 1 and 2, respectively, with mean durations of 31 and 26 minutes, respectively, and a mean distance of catheter insertion of 11.9 cm. Maxillary artery occlusion was successful in five of seven arteries (phase 1), with a mean surgical duration of 47 minutes and a mean distance of catheter insertion of 42.8 cm. CONCLUSION: Balloon catheter occlusion of the ICA, ECA, and MA was achieved in most standing horses. CLINICAL SIGNIFICANCE: Balloon catheter occlusion in standing horses provides an alternative to prevent or treat hemorrhage related to guttural pouch mycosis, particularly in horses in which general anesthesia might pose a risk.

Dehydrojuncusol, a Natural Phenanthrene Compound Extracted from Juncus maritimus, Is a New Inhibitor of Hepatitis C Virus RNA Replication.

Recent emergence of direct-acting antivirals (DAAs) targeting hepatitis C virus (HCV) proteins has considerably enhanced the success of antiviral therapy. However, the appearance of DAA-resistant-associated variants is a cause of treatment failure, and the high cost of DAAs renders the therapy not accessible in countries with inadequate medical infrastructures. Therefore, the search for new inhibitors with a lower cost of production should be pursued. In this context, the crude extract of Juncus maritimus Lam. was shown to exhibit high antiviral activity against HCV in cell culture. Bio-guided fractionation allowed the isolation and identification of the active compound, dehydrojuncusol. A time-of-addition assay showed that dehydrojuncusol significantly inhibited HCV infection when added after virus inoculation of HCV genotype 2a (50% effective concentration [EC50] = 1.35 µM). This antiviral activity was confirmed with an HCV subgenomic replicon, and no effect on HCV pseudoparticle entry was observed. Antiviral activity of dehydrojuncusol was also demonstrated in primary human hepatocytes. No in vitro toxicity was observed at active concentrations. Dehydrojuncusol is also efficient on HCV genotype 3a and can be used in combination with sofosbuvir. Interestingly, dehydrojuncusol was able to inhibit RNA replication of two frequent daclatasvir-resistant mutants (L31M or Y93H in NS5A). Finally, mutants resistant to dehydrojuncusol were obtained and showed that the HCV NS5A protein is the target of the molecule. In conclusion, dehydrojuncusol, a natural compound extracted from J. maritimus, inhibits infection of different HCV genotypes by targeting the NS5A protein and is active against resistant HCV variants frequently found in patients with treatment failure.IMPORTANCE Tens of millions of people are infected with hepatitis C virus (HCV) worldwide. Recently marketed direct-acting antivirals (DAAs) targeting HCV proteins have enhanced the efficacy of treatment. However, due to its high cost, this new therapy is not accessible to the vast majority of infected patients. Furthermore, treatment failures have also been reported due to the appearance of viral resistance. Here, we report on the identification of a new HCV inhibitor, dehydrojuncusol, that targets HCV NS5A and is able to inhibit RNA replication of replicons harboring resistance mutations to anti-NS5A DAAs used in current therapy. Dehydrojuncusol is a natural compound isolated from Juncus maritimus, a halophilic plant species that is very common in coastlines worldwide. This molecule might serve as a lead for the development of a new therapy that is more accessible to hepatitis C patients in the future.

Antimicrobial and Light-Enhanced Antimicrobial Activities, Cytotoxicity and Chemical Variability of All Tunisian Eryngium Species.

This study was performed on all Eryngium species growing in Tunisia in order to evaluate their intra and interspecies variabilities and to investigate their biological activities. These species are used in traditional medicine, and literature about the phytochemical investigations of most of them is scarce. Antimicrobial and light-enhanced activities were tested against multiresistant microorganisms and extended spectrum beta-lactamase producing bacteria (ESBL). All studied species showed antimicrobial effect with several MIC values lower than 70 µg/ml. Tested Eryngium species have proven to be a promising source of photoactive compounds, while light-enhanced activity offers an alternative for the inactivation of pathogenic microorganisms which is currently subjected to a great interest. This is the first report of this activity in genus Eryngium. A significant improvement of antimicrobial activity with UV irradiation was observed, mainly for E. dichotomum, E. ilicifolium and E. triquetrum. Cytotoxicity, studied for the first time for the most species, was evaluated against cancer (J774) and non-cancer (WI38) human cell lines. Chemical composition of volatile compounds presented in the most active crude extracts (petroleum ether extracts) of the aerial parts was investigated using GC/MS analysis and was submitted to statistical analyses. It revealed their high content of bioactive phytochemicals, particularly oxygenated sesquiterpenes like spathulenol, ledol and α-bisabolol but also hydrocarbon sesquiterpenes such as ß-bisabolene and copaene, as well as polyacetylene derivatives such as falcarinol. Statistical analyses permitted to evaluate the interrelations between all Tunisian Eryngium species.

Phenolic Compounds from Humulus lupulus as Natural Antimicrobial Products: New Weapons in the Fight against Methicillin Resistant Staphylococcus aureus, Leishmania mexicana and Trypanosoma brucei Strains.

New anti-infective agents are urgently needed to fight microbial resistance. Methicillin-resistant Staphylococcus aureus (MRSA) strains are particularly responsible for complicated pathologies that are difficult to treat due to their virulence and the formation of persistent biofilms forming a complex protecting shell. Parasitic infections caused by Trypanosoma brucei and Leishmania mexicana are also of global concern, because of the mortality due to the low number of safe and effective treatments. Female inflorescences of hop produce specialized metabolites known for their antimicrobial effects but underexploited to fight against drug-resistant microorganisms. In this study, we assessed the antimicrobial potential of phenolic compounds against MRSA clinical isolates, T. brucei and L. mexicana. By fractionation process, we purified the major prenylated chalcones and acylphloroglucinols, which were quantified by UHPLC-UV in different plant parts, showing their higher content in the active flowers extract. Their potent antibacterial action (MIC < 1 µg/mL for the most active compound) was demonstrated against MRSA strains, through kill curves, post-antibiotic effects, anti-biofilm assays and synergy studies with antibiotics. An antiparasitic activity was also shown for some purified compounds, particularly on T. brucei (IC50 < 1 to 11 µg/mL). Their cytotoxic activity was assessed both on cancer and non-cancer human cell lines.

Neoboutonia melleri var velutina Prain: in vitro and in vivo hepatoprotective effects of the aqueous stem bark extract on acute hepatitis models.

BACKGROUND: Hepatitis is a liver inflammation caused by different agents and remains a public health problem worldwide. Medicinal plants are an important source of new molecules being considered for treatment of this disease. Our work aims at evaluating the hepatoprotective properties of Neoboutonia velutina, a Cameroonian medicinal plant. METHODS: The aqueous extract has been prepared using phytochemical methods. HepG2 cells were used to assess anti-inflammatory properties of the extract at different concentrations. Acute hepatitis models (Carbon tetrachloride and Concanavalin A) were performed in mice receiving or not receiving, different extract doses by gavage. Liver injury was assessed using histology, transaminases and pro-inflammatory markers. Extract antioxidant and radical scavenging capacities were evaluated. RESULTS: The extract led to a significant decrease in pro-inflammatory cytokine expression in vitro and to a remarkable protection of mice from carbon tetrachloride-induced liver injury, as shown by a significant decrease in dose-dependent transaminases level. Upon extract treatment, inflammatory markers were significantly decreased and liver injuries were limited as well. In the Concanavalin A model, the extract displayed weak effects. CONCLUSIONS: Taking into account underlying mechanisms in both hepatitis models, we demonstrate the extract's radical scavenging capacity. Neoboutonia velutina displays a potent hepatoprotective effect mediated through radical scavenging properties.

Ex vivo biomechanical stability of 5 cricoid-suture constructs for equine laryngoplasty.

OBJECTIVE: To determine the biomechanical properties of 5 suture constructs in the equine cricoid under cyclic loading and load to failure testing. STUDY DESIGN: Ex vivo study. SAMPLES: Seventy-five equine cadaver larynges. METHODS: Each larynx was implanted with 1 of 5 cricoid-suture constructs. The standard laryngoplasty, where a suture is passed once through the cricoid, including its caudal edge, was used in 2 constructs: 1 with 5 USP Ethibond (ES) and 1 with 2 mm Fibertape (FS). In the third construct, the 2 mm Fibertape was passed twice through the cricoid including its caudal edge (Double Loop-DL). Constructs 4 and 5 used 2 mm Fibertape in a U-shaped loop passed through the cricoid but excluding its caudal edge. One construct was supported with a metallic button (MB) on the caudo-ventral aspect of the cricoid while the other included only the U-shaped loop (U). Constructs were subjected to cyclic loading and to single cycle to failure. Reduction of the left-to-right arytenoid angle quotient (LRQ), suture migration, and load at failure were compared. RESULTS: LRQ reduction after cyclic loading was lower in MB and U than ES constructs. During cyclic loading, suture migration was reduced in MB, U, and DL compared to ES constructs. Mean load at failure was lower in FS and U than in ES constructs. CONCLUSION: Loss of abduction after equine laryngoplasty may be reduced and pullout forces increased by applying a MB construct in the cricoid cartilage. In vivo testing is required to verify these results.

Chemical Composition and Antimicrobial Activity of the Essential Oil from Aerial Parts and Roots of Eryngium barrelieri Boiss. and Eryngium glomeratum Lam. from Tunisia.

The study of chemical composition and biological activity of unexplored essential oils may open new perspectives on their potential use in facing major health concerns such as drug-resistant infections. The present study investigates the chemical composition and antimicrobial effects of previously unstudied essential oils obtained from genus Eryngium: Eryngium glomeratum Lam. and Eryngium barrelieri Boiss. The chemical compositions of the essential oils from aerial parts and roots of both species were studied using GC and GC/MS analytical technics. The analysis led to the identification of 102 compounds totalizing 85 - 94% of all detected compounds. Essential oils were characterized by the predominance of oxygenated sesquiterpenes. The oils obtained from aerial parts were tested against 36 microbial strains by agar dilution method and showed minimum inhibitory concentrations (MIC) in the range of 2 - 625 µg/ml. A strong antibacterial activity against multiresistant Pseudomonas aeruginosa was observed especially from E. glomeratum essential oil with MIC value up to 2 µg/ml. These findings give significant information about the pharmacological activity of these essential oils, which suggest their potential use to develop new remedies, or as sources of active compounds.

Internal Fixation of Cervical Fractures in Three Horses.

OBJECTIVE: To describe the surgical treatment outcome of cervical fractures in 3 horses. STUDY DESIGN: Case report. ANIMALS: Three client-owned horses with cervical vertebral fractures. METHODS: Three horses were refered for neck stiffness, pain, and ataxia after a cervical trauma because of a fall. Radiographic examination showed an oblique displaced fracture of the caudal aspect of the body of the second cervical vertebra (C2) in horse 1, an oblique displaced fracture of the caudal aspect of C4 involving the disc between C4 and C5 in horse 2, and a displaced transverse fracture of the body of the axis (C2) extending to the lateral arches and involving the vertebral canal in horse 3. In horse 1, the fracture was reduced and stabilized using a 14-hole narrow DCP plate, applied ventrally, and fixed with cancellous screws. A cervical fusion was performed. In horses 2 and 3, fracture fixation was performed using a 5-hole narrow LCP and 5 mm locking screws. RESULTS: All horses showed improvement and returned to full activity. The fracture healed in all horses. CONCLUSION: Internal fixation of cervical fracture in these horses was associated with minimal complications, and was associated with healing and a highly functional outcome in all horses. The LCP was preferred and would be recommended for ventral stabilization of selected cases of vertebral fractures.

Mechanical comparison of 3 anchoring techniques in the muscular process for laryngoplasty in the equine larynx.

OBJECTIVE: To compare mechanical properties of 2 techniques with a conventional technique for anchoring the muscular process in a laryngoplasty procedure. STUDY DESIGN: Experimental ex vivo study. SAMPLE POPULATION: Equine larynges (n = 60). METHODS: A single loop (SL), a screw (SC), and a double loop technique (DL) were compared. Constructs were subjected to cyclic loading, oscillating from 5 to 50 N for 3000 cycles, followed by a single cycle to failure test. Mean distraction, load at failure, stiffness, and failure mode were compared between groups. RESULTS: Mean ± SD distraction in cyclic loading was greater for DL (2.1 ± 0.7 mm) than for SL (1.9 ± 1.3 mm) and SC (1.539 ± 0.9 mm); however, there was no significant difference between SL and SC or between SL and DL. Mean ultimate failure load was greater for DL (240 ± 44.56 N) than for SC (189.59 ± 46.16 N) and SL (150.93 ± 44.43 N) and greater for SC compared with SL. Failure occurred by cartilage tearing for DL and SL, and by screw pull out (n = 13) or knot slippage (4) for SC. CONCLUSION: In cyclic loading, SC is more stable than DL and at least as stable as SL. In single cycle to failure, DL is the strongest construct and SC is stronger than SL.

Standing laparoscopic inguinal hernioplasty using cyanoacrylate for preventing recurrence of acquired strangulated inguinal herniation in 10 stallions.

OBJECTIVE: To describe a technique for standing inguinal hernioplasty in horses using cyanoacrylate glue, and to evaluate its effect on prevention of recurrent inguinal herniation in stallions that had previous acquired strangulated inguinal hernia (SIH). STUDY DESIGN: Case series. ANIMALS: Stallions (n = 10) with a history of SIH. METHODS: Hernioplasty was performed in standing horses using 4 laparoscopic portals. The mesorchium was retracted caudomedially using Babcock forceps. A flexible polyethylene extension tube was introduced through the sheath of a laparoscopic needle and n-butyl-2-cyanoacrylate (2 mL) was injected into the inguinal canal including its margins while a 2nd Babcock forceps prevented deep ventral diffusion of the cyanoacrylate. The craniolateral parts of the vaginal ring were compressed until full adhesion between the visceral and parietal walls was achieved. In 2 horses, the lateral part of the vaginal ring was sutured before gluing. A contralateral approach was used to check the caudomedial part of the vaginal ring. RESULTS: No recurrence (1-4 years) of inguinal hernia was reported. No major complications occurred and cosmetic outcome was excellent. All horses were used for their intended purpose and 7 horses being used as breeding stallions remained fertile. CONCLUSION: Standing inguinal hernioplasty using cyanoacrylate seems to provide efficient and secure closure of the vaginal ring in stallions.

Two-step surgery combining standing laparoscopy with recumbent ventral median celiotomy for removal of enlarged pathologic ovaries in 20 mares.

OBJECTIVES: To report a combination of standing laparoscopic technique for intra-abdominal dissection of the mesovarial pedicle followed by a limited ventral median celiotomy under general anesthesia for removal of enlarged ovaries in mares. STUDY DESIGN: Case series. ANIMALS: Mares (n = 20) aged 3-22 years with unilateral enlarged ovaries. METHODS: Enlarged ovaries were confirmed by transrectal palpation and ultrasonography. After sedation, 3 laparoscopic portals were made in the paralumbar fossa. The mesovarium was desensitized and dissected using a vessel sealing device, and the ovary was left free in the abdomen. Then under general anesthesia, the mare was positioned in dorsal recumbency and an 8 cm ventral median celiotomy made for ovary retrieval in a specimen bag. RESULTS: This 2-step procedure was successfully used for removal enlarged ovaries (12-50 cm) in 17 mares and for management of behavioral problems in 3 mares. No operative or postoperative complications occurred. Owner satisfaction and cosmesis were considered excellent. CONCLUSIONS: Standing laparoscopy combined with a limited median celiotomy is a safe technique for ovariectomy in mares. This technique mitigates most of the disadvantages of standing flank ovariectomy and a conventional open ventral median approach.

Standing gustatory papillae biopsy procedure for antemortem diagnosis of equine grass sickness.

OBJECTIVE: Diagnosing equine grass sickness (EGS) requires histopathological evidence of chromatolysis and/or neuronal loss in peripheral autonomic ganglia. Previous investigators performed postmortem biopsies of gustatory papillae located on the tongue and found chromatolytic subgemmal neurons in all 13 EGS horses. The present study aimed to design a standardized lingual biopsy sampling method through a transbuccal approach in healthy standing horses and assess the quality of the obtained samples, to allow antemortem diagnosis of EGS in clinical cases. ANIMALS: 6 healthy horses. METHODS: A transbuccal approach was performed bilaterally in 6 healthy standing horses. After having reached a deep level of sedation, horses were placed in stocks and a Günther mouth gag was inserted. Local anesthesia followed by a vertical full thickness incision was performed on both cheeks. Foliate papillae biopsies were carried out using an arthroscopic rongeur inserted through each incision site under oral endoscopic control. Tongue movements were restricted with diazepam. Histological assessment of taste buds and subgemmal plexi neurons was performed using H&E-stained longitudinal sections. RESULTS: The procedure was well tolerated in all horses. Minor complications observed were a transient facial paralysis, some incisional fluid collection, and abscesses. Ten samples (10/12) were suitable for assessment of neuronal perikarya. CLINICAL RELEVANCE: This procedure was safe for subgemmal plexus biopsy in healthy standing horses. The obtained samples were adequate as long as they were neatly cut lengthwise for inclusion. The technique was also used for 2 clinical cases and revealed the complete absence of neuronal perikarya, confirming chronic EGS.

Antibacterial and anti-coronavirus investigation of selected Senegalese plant species according to an ethnobotanical survey.

ETHNOPHARMACOLOGICAL RELEVANCE: In Senegal, upper and lower respiratory tract infections constitute a real health problem. To manage these disorders, most people rely on the use of local medicinal plants. This is particularly the case for species belonging to the botanical families, Combretaceae, Fabaceae, Myrtaceae and Rubiaceae, which are widely used to treat various respiratory problems such as colds, flu, rhinitis, sinusitis, otitis, angina, bronchitis, bronchiolitis and also pneumonia. AIM OF THE STUDY: The aim of this study was to identify medicinal plants traditionally used for the management of infectious diseases, in particular those of the respiratory tract. On the basis of these ethnopharmacological uses, this study made it possible to highlight the antibacterial, antiviral and cytotoxic activities of selected plant species. MATERIALS AND METHODS: An ethnobotanical survey was conducted in Senegal among informants, including herbalists, traditional healers, and households, using medicinal plants in the management of infectious diseases, with a focus on respiratory tract infections. The most cited plant species were evaluated in vitro on a panel of 18 human pathogenic bacteria may be involved in respiratory infections and against the human coronavirus HCoV-229E in Huh-7 cells. The antiviral activity of the most active extracts against HCoV-229E was also evaluated on COVID-19 causing agent, SARS-CoV-2 in Vero-81 cells. In parallel, cytotoxic activities were evaluated on Huh-7 cells. RESULTS: A total of 127 informants, including 100 men (78.74%) and 27 women (21.26%) participated in this study. The ethnobotanical survey led to the inventory of 41 plant species belonging to 19 botanical families used by herbalists and/or traditional healers and some households to treat infectious diseases, with a specific focus on upper respiratory tract disorders. Among the 41 plant species, the most frequently mentioned in the survey were Guiera senegalensis J.F. Gmel. (95.2%), Combretum glutinosum Perr. Ex DC. (93.9%) and Eucalyptus spp. (82.8%). Combretaceae (30.2%) represented the most cited botanical family with six species, followed by Fabaceae (29.3%, 12 species). A total of 33 crude methanolic extracts of the 24 plant species selected for their number of citations were evaluated in vitro for their antimicrobial and cytotoxic activities. Guiera senegalensis, Combretum glutinosum, Vachellia nilotica subsp. tomentosa (Benth.) Kyal. & Boatwr, Eucalyptus camaldulensis Dehnh., and Terminalia avicennioides Guill. & Perr., showed antibacterial activities. The most active plants against HCoV-229E were: Ficus sycomorus L., Mitragyna inermis (Willd.) Kuntze, Pterocarpus erinaceus Poir., and Spermacoce verticillata L. One of these plants, Mitragyna inermis, was also active against SARS-CoV-2. CONCLUSION: This work confirmed the anti-infective properties of plant species traditionally used in Senegal. Overall, the most frequently cited plant species showed the best antibacterial activities. Moreover, some of the selected plant species could be considered as a potential source for the management of coronavirus infections. This new scientific data justified the use of these plants in the management of some infectious pathologies, especially those of the respiratory tract.

Anthocyanin phytochemical profiles and anti-oxidant activities of Vitis candicans and Vitis doaniana.

INTRODUCTION: Grapes are one of the most important fruit crops in the world. The quality of red grape berries greatly depends on skin colour, mainly due to the anthocyanin profile. Today, the American Vitis species have the greatest potential for breeding work. They have multiple resistance properties in comparison with Vitis vinifera but little is known about their anthocyanin content. OBJECTIVE: To determine the anti-oxidant properties and anthocyanin profile of two American species, Vitis candicans and Vitis doaniana, by using LC-MS(n) and LC-NMR. METHODS: Grape extracts were prepared by extraction of berry skins with acidified methanol. The complete structure elucidation of the individual anthocyanins was performed with LC-MS(n) , LC-NMR and NMR experiments. Individual anthocyanins in the extracts were quantified by using malvidin glucoside as external standard. The anti-oxidant activities of grape skin extracts were evaluated by using 1,1-diphenyl-2-picrylhydrazyl (DPPH•) radical scavenging and oxygen radical absorbance capacity (ORAC) assays. RESULTS: By using LC-MS(n) and LC-NMR experiments, 30 anthocyanins were identified and quantified in the two Vitis species, including two new cis-p-coumaroyl derivatives. Vitis candicans and V. doaniana showed significant differences in their anthocyanin profile. These two Vitis species possess low-to-medium anti-oxidant activities in comparison with V. vinifera. CONCLUSION: The profiles of 30 anthocyanins were established unambiguously in two American Vitis species.

Antifungal Activity of Hop Leaf Extracts and Xanthohumol on Two Strains of Venturia inaequalis with Different Sensitivities to Triazoles.

Hop cones are well-known for their antimicrobial properties, attributed to their specialized metabolites. Thus, this study aimed to determine the in vitro antifungal activity of different hop parts, including by-products such as leaves and stems, and some metabolites against Venturia inaequalis, the causal agent of apple scab. For each plant part, two types of extracts, a crude hydro-ethanolic extract and a dichloromethane sub-extract, were tested on spore germination of two strains with different sensitivities to triazole fungicides. Both extracts of cones, leaves and stems were able to inhibit the two strains, whereas rhizomes did not show activity. The apolar sub-extract of leaves appeared as the most active modality tested with half maximal inhibitory concentrations (IC50) of 5 and 10.5 mg·L-1 on the sensitive strain and the strain with reduced sensitivity, respectively. Differences in activity level between strains were noticed for all active modalities tested. Sub-extracts of leaves were then separated into seven fractions by preparative HPLC and tested on V. inaequalis. One fraction, containing xanthohumol, was especially active on both strains. This prenylated chalcone was then purified by preparative HPLC and showed significant activity against both strains, with IC50 of 1.6 and 5.1 mg·L-1. Therefore, xanthohumol seems to be a promising compound to control V. inaequalis.

Laryngeal reinnervation using the spinal accessory nerve: Electromyographic study of the sternomandibularis muscle.

BACKGROUND: Selective laryngeal reinnervation using the first and second cervical nerve (C1C2) is a treatment option for recurrent laryngeal neuropathy that aims to restore the function of the cricoarytenoideus dorsalis (CAD) muscle. Despite the technique's satisfying success rate, it has several limitations. These triggered the search for another potential donor nerve that could reduce CAD muscle fatigue and shorten rehabilitation. The ventral branch of the spinal accessory nerve, providing motor innervation to the sternomandibularis (SM) muscle, was identified as a potentially well-adapted nerve. OBJECTIVES: To gather normative data from SM muscle activity induced by physiological spinal accessory nerve stimulation at rest and during exercise, and to determine SM muscle activity and fibre type recruitment relative to posture, gait and respiratory cycle. STUDY DESIGN: Clinical observational study. METHODS: Surface electromyography (EMG) of the SM muscle was performed in nine horses: trained warmbloods, Thoroughbreds and Standardbreds. Signals were recorded in different feeding postures and at exercise, which included standardised treadmill exercise tests, lungeing and ridden work. RESULTS: Timing of the SM muscle contraction coincided with inspiration at gallop. Intra-individual mean SM muscle activity increased with exercising speed (four times higher in Thoroughbreds at gallop than walk and seven times higher in Standardbreds at high-speed trot than walk). Moreover, the SM muscle was strongly activated at rest when the horse was grazing (seven times more than when it was eating out of a hay net or stable feeder). Frequency domain analysis revealed a predominant type I muscle fibre recruitment during feeding and at exercise (type I muscle fibre activity was at least 2.5 times higher than type II muscle fibre activity). MAIN LIMITATIONS: Lack of reference data on equine EMG. CONCLUSIONS: This study confirmed the potentially advantageous properties of the spinal accessory nerve for laryngeal reinnervation.

Multivariate analysis of chemical and genetic diversity of wild Humulus lupulus L. (hop) collected in situ in northern France.

The hop plant (Humulus lupulus L.) has been exploited for a long time for both its brewing and medicinal uses, due in particular to its specific chemical composition. These last years, hop cultivation that was in decline has been experiencing a renewal for several reasons, such as a craze for strongly hopped aromatic beers. In this context, the present work aims at investigating the genetic and chemical diversity of fifty wild hops collected from different locations in Northern France. These wild hops were compared to ten commercial varieties and three heirloom varieties cultivated in the same sampled geographical area. Genetic analysis relying on genome fingerprinting using 11 microsatellite markers showed a high level of diversity. A total of 56 alleles were determined with an average of 10.9 alleles per locus and assessed a significant population structure (mean pairwise FST = 0.29). Phytochemical characterization of hops was based on volatile compound analysis by HS-SPME GC-MS, quantification of the main prenylated phenolic compounds by UHPLC-UV as well as untargeted metabolomics by UHPLC-HRMS and revealed a high level of chemical diversity among the assessed wild accessions. In particular, analysis of volatile compounds revealed the presence of some minor but original compounds, such as aromadendrene, allo-aromadendrene, isoledene, ß-guaiene, α-ylangene and ß-pinene in some wild accessions; while analysis of phenolic compounds showed high content of ß-acids in these wild accessions, up to 2.37% of colupulone. Genetic diversity of wild hops previously observed was hence supported by their chemical diversity. Sample soil analysis was also performed to get a pedological classification of these different collection sites. Results of the multivariate statistical analysis suggest that wild hops constitute a huge pool of chemical and genetic diversity of this species.

Natural stilbenoids: distribution in the plant kingdom and chemotaxonomic interest in Vitaceae.

Stilbenoids, a family of polyphenols known for the complexity of their structure and for their diverse biological activities, occur with a limited but heterogeneous distribution in the plant kingdom. The most prominent stilbene containing plant family, the Vitaceae, represented by the famous wine producing grape vines Vitis vinifera L., is one of the richest sources of novel stilbenes currently known, together with other families, such as Dipterocarpaceae, Gnetaceae and Fabaceae. This review focuses on the distribution of stilbenes and 2-arylbenzofuran derivatives in the plant kingdom, the chemical structure of stilbenes in the Vitaceae family and their taxonomic implication.