Apigenin inhibits platelet adhesion and thrombus formation and synergizes with aspirin in the suppression of the arachidonic acid pathway

Previous studies using washed platelets demonstrated that certain flavonoids inhibit platelet function through several mechanisms including blockade of TxA(2) receptors (TPs). We aimed to analyze the binding capacity of flavonoids to TPs in platelet rich plasma (PRP), investigated their effect in flowing blood, and evaluated the ability of apigenin to improve the efficacy of aspirin in the inhibition of platelet aggregation. The binding of flavonoids to TPs in PRP was explored using binding assays and the TP antagonist [ (3)H]SQ29548. Effects of flavonoids on platelet adhesion were assessed using arterial subendothelium with annular plate perfusion chambers, and global evaluation of apigenin on high-shear-dependent platelet function was determined by the PFA-100. To evaluate the ability of apigenin to potentiate the effect of aspirin, arachidonic acid-induced platelet aggregation was measured prior to and after consumption of subaggregatory doses of aspirin in the presence or absence of apigenin. Binding assays revealed that apigenin was an efficient competitor of [ (3)H]SQ29548 binding to PRP ( K i = 155.3 +/- 65.4 microM), and perfusion studies showed that apigenin, genistein, and catechin significantly diminished thrombus formation when compared to control (26.2 +/- 3.8, 33.1 +/- 5.2, and 26.2 +/- 5.2 vs 76.6 +/- 2.6%, respectively; p < 0.05). Apigenin, similarly to the TP antagonist SQ29548, significantly prolonged collagen epinephrine-induced PFA-100 closure time in comparison to the control and, when added to platelets that had been exposed in vivo to aspirin, potentiated its inhibitory effect on platelet aggregation. The inhibitory effect of some flavonoids in the presence of plasma, particularly apigenin, might in part rely on TxA(2) receptor antagonism. There is a clear increase in the ex vivo antiplatelet effect of aspirin in the presence of apigenin, which encourages the idea of the combined use of aspirin and certain flavonoids in patients in which aspirin fails to properly suppress the TxA(2) pathway.     

A Preliminary Study on the Effects of Dietary Energy and Melatonin on the Ex Vivo Production of Progesterone and Prostaglandin F2α by the Corpora Lutea and Endometrial Tissue of Ewes

Veterinary Research Communications -     

White spot syndrome virus (WSSV) infection and immunity responses in white shrimp (Litopenaeus vannamei) exposed to sublethal levels of metals

To determine if exposure to a sublethal mixture of metals (Cd, Cu, Fe, Mn, Pb and Zn) increases susceptibility to White spot syndrome virus (WSSV) infection, Litopenaeus vannamei juveniles were fed WSSV‐infected shrimp tissues after 21 days of exposure to the metal mixture (WS‐MM treatment). Other treatments consisted of shrimp not exposed to metals and fed infected tissues (WS), and shrimp fed healthy tissues and exposed (MM) or not exposed to metals (C). The presence of viral DNA and inclusion bodies was detected at 32 hr postinfection (hpi) in the stomach epithelium of shrimp from the WS treatment, and eight hours later in shrimp from the WS‐MM treatment, possibly because of an initial negative effect of metals in viral replication. At 40 hpi, the severity of infection represented by the histopathological index increased in both WS and WS‐MM treatments, and values were higher in WS‐MM than in WS shrimp at the end of the experiment. From 56 hpi to the end of experiment, total hemocyte counts were lower in both WS‐MM and WS treatments, and concentrations were particularly low in WS‐MM shrimp. Conversely, phenoloxidase activity was higher in the WS‐MM treatment from 32 to 56 hpi, suggesting a possible role of the prophenoloxidase (proPO) system in the antiviral defense against WSSV. The presence of heavy metals at sublethal concentrations may increase shrimp susceptibility to WSSV; this is supported by a decrease in circulating hemocytes, an increase in the humoral response, and the development of a higher number of WSSV inclusion bodies.     

Limited evidence for genetic variation for resistance to the white spot syndrome virus in Indian populations of Penaeus monodon

There has been a highly detrimental impact of the white spot syndrome virus (WSSV) on black tiger shrimp (Penaeus monodon) aquaculture in India. Currently, no cost‐effective measures are available for controlling the disease. One alternative is to improve WSSV resistance through a selective breeding programme for disease‐resistant shrimp, provided that genetic variation exists for this trait. The aim of this study was to evaluate the evidence for genetic variation in resistance to WSSV in P. monodon sourced from Indian populations. Post‐larval shrimp (n=1950) from 54 full‐sibling families were challenged with WSSV using WSSV‐infected mince meat. The heritability was estimated using four different statistical models fitted to the resulting time to death data, including two linear models and two Weibull proportional hazard frailty models. None of the estimated heritabilities were significantly different from zero. We suggest three possible explanations for these results: there actually is very little variation between P. monodon in WSSV resistance and all individuals are highly susceptible to the disease; there is genetic variation in resistance to WSSV in P. monodon but we did not find it in our experiment because the level of challenge in the experiment was too high to allow genetic differences to be expressed; the variation is due to mutations conferring resistance, which are at a low frequency in the population, and we did not sample a broad enough genetic base to capture these mutations.     

Phenolic acid and flavonol water extracts of Delonix regia red flowers

Traditional healthy beverages in several African counties were home-made by water extraction of flowers of Delonix regia (Boj.) Raf. This tree belongs to the local biodiversity of plants used in the local medicine. It is also part of the traditional bioproducts marketed within African village communities. Scaling-up at pilot plant level, by mimicking this local recipe, lead to concentrated bioproducts containing various natural phenolic compounds, most of them have never been yet published. A three-steps process was used to prepare concentrated of water-extracted soluble phenolics. This process included: phenol extraction of dried plant by overnight diffusion in acidified water, clarification then concentration of the macerate, using membrane technology. Total phenol contents were determined as gallic acid equivalents using the Folin-Ciocalteu method. Total flavonoid contents, as delphinidin equivalents, were determined using the UV-vis colorimetric method. Comparison of concentration factors of both phenol families (phenolic acids and flavonols) contained in the 3 co-products obtained during the process were made between crude extracts, cross-flow microfiltration extracts and reverse osmosis concentrated extracts. Individual phenol compounds found in the products obtained were characterized and quantified by HPLC-DAD and by HPLC-MS². Among the 11 phenol compounds identified in the water-extracts, 3 have been already identified in the literature (2 phenolic acids and 1 flavonol), and 8 new compounds, in noticeable amounts, have never been cited. We have confirmed the presence of phenolic acids such as 3,4,5-trihydroxybenzoic (gallic acid), 3,4-dihydroxybenzoic (protocatechuic acid) and of quercetin. We have characterized for the first time the presence of 2-hydroxy 5-[(3,4,5 trihydroxyphenyl) carbonyl oxy] benzoic acid and of 7 other flavonols belonging to the quercetin, kaempferol and isorhamnetol compound families. Using both LCMS and NMR analyses, they have been identified and quantified as: rutin, quercetin 3-O-glucoside, quercetin 3-O-galactoside, quercetin trihexoside, quercetin 3-O-robinobioside, kaempferol rhamnosylhexoside and isorhamnetol rhamnosyl-hexoside.     

Instrumental and sensory characterization of heat-induced odorants in aseptically packaged soy milk

Predominant heat-induced odorants generated in soy milk by ultrahigh-temperature (UHT) processing were evaluated by sensory and instrumental techniques. Soy milks processed by UHT (143 degrees C/14 s, 143 degrees C/59 s, 154 degrees C/29 s) were compared to a control soy milk (90 degrees C/10 min) after 0, 1, and 7 days of storage (4.4 +/- 1 degrees C). Dynamic headspace dilution analysis (DHDA) and solvent-assisted flavor evaporation (SAFE) in conjunction with GC-olfactometry (GCO)/aroma extract dilution techniques and GC-MS were used to identify and quantify major aroma-active compounds. Sensory results revealed that intensities of overall aroma and sulfur and sweet aromatic flavors were affected by the processing conditions. Odorants mainly responsible for the changes in sulfur perception were methional, methanethiol, and dimethyl sulfide. Increases in 2-acetyl-1-pyrroline, 2-acetyl-thiazole, and 2-acetyl-2-thiazoline intensities were associated with roasted aromas. A marginal increase in intensity of sweet aromatic flavor could be explained by increases in 2,3-butanedione, 3-hydroxy-2-butanone, beta-damascenone, and 2- and 3-methylbutanal. Predominant lipid-derived odorants, including (E,E)-2,4-nonadienal, (E,E)-2,4-decadienal, (E,Z)-2,4-decadienal, (E)-2-nonenal, (E)-2-octenal, 1-octen-3-one, 1-octen-3-ol, and (E,Z)-2,6-nonadienal, were affected by processing conditions. Intensities of overall aroma and sulfur notes in soy milk decreased during storage, whereas other sensory attributes did not change. Color changes, evaluated by using a Chroma-meter, indicated all UHT heating conditions used in this study generated a more yellow and saturated color in soy milk in comparison to the control soy milk.     

Episodic recruitment of young Prochilodus lineatus (Valenciennes, 1836) (Characiformes: Prochilodontidae) during high discharge in a floodplain lake of the River Paraná, Argentina

Prochilodus lineatus (Val.) is one of the main target species of South American freshwater fisheries. The following null hypotheses regarding juvenile recruitment of P. lineatus were tested: (a) recruitment is not determined by variations in larval abundance and (b) recruitment is not determined by variations in discharge. For this purpose, variations in abundance of larvae in main channel drift and monthly captures of juveniles in a floodplain lake of the River Paraná were examined weekly between 2009 and 2016. Mean annual abundances of larval densities and CPUE of juveniles were correlated with a set of hydrometric variables. A positive correlation was found between the abundance of juveniles with high and persistent flood pulses. By contrast, larval abundance was not correlated with juvenile abundance. Pronounced contrast was found in the recruitment of P. lineatus between years of high and low discharge, which supports the hypothesis that floods are the main determining factor for recruitment of this species.     

Bilateral epididymal sperm granulomas following urethrostomy in a German shepherd dog

Bilateral enlargement of both epididymes was observed in a 6-year-old German shepherd dog following a pre-scrotal urethrostomy. Testicular parenchyma showed regular structure, and the spermatogenesis and the steroidogenic functions were not modified. However, macroscopic examination of the tail and the body of both epididymes exhibited multiple white and well-delimited foci. Histopathological study of the epididymes confirmed the development of granulomas associated with extravasated spermatozoa. Urethrostomy caused a severe stenosis of the penile urethra, favouring the retention of urine at the urinary bladder. The retrograde pressure exerted by the distension of the urinary bladder could have allowed the urine to reach the prostatic urethra and the deferent ducts and, finally, the epididymes, causing irritation and rupture of the mucous layer of the epididymal duct, the consequent sperm extravasation and the development of sperm granulomas. We speculate that the inadequate surgical resolution of the urethral calculi caused the bladder distension, the subsequent retrograde flow of urine and the development of the lesions.     

Components of early competition between upland rice (Oryza sativa L.) and Brachiaria brizantha (Hochst. ex A.Rich) Stapf.

Abstract

In Latin America early weed competition is a constraint upon upland rice production, leading to intensive herbicide use. Breeding for rice competitiveness may help reduce the need for herbicides. Two consecutive screenhouse pot experiments were conducted with 13 upland cultivars grown alone or in mixtures with Brachana brizantha to identify plant characteristics for early rice competitiveness which could serve as selection criteria. Rice growth reduction from B. brizantha competition was evident in all cultivars by 30 d.a.e., and the traits most related to cultivar competitiveness were: total above‐ground plant biomass > leaf weight > stem weight > leaf area. Plant height and tillering were least affected by competition. Rice and B. brizantha partially avoided competition since the competing canopies did not fully intercept the incoming light. Traits measured in monoculture did not correlate well with the competition‐induced growth reductions. In breeding for competitiveness, selection according to the above traits should be conducted on rice growing in competition, rather than in monoculture. A measure of plant biomass or leaf area at 30 d.a.e. appears to provide the most convenient estimates of early interference.     

A comparison of the duration and quality of recovery from isoflurane, sevoflurane and desflurane anaesthesia in dogs undergoing magnetic resonance imaging

ObjectiveTo compare the recovery after anaesthesia with isoflurane, sevoflurane and desflurane in dogs undergoing magnetic resonance imaging (MRI) of the brain.Study designProspective, randomized clinical trial.AnimalsThirty‐eight dogs weighing 23.7 ± 12.6 kg.MethodsFollowing pre‐medication with meperidine, 3 mg kg^?1 administered intramuscularly, anaesthesia was induced intravenously with propofol (mean dose 4.26 ± 1.3 mg kg^?1), the trachea was intubated, and an inhalational anaesthetic agent was administered in oxygen. The dogs were randomly allocated to one of three groups: group I (n = 13) received isoflurane, group S (n = 12) received sevoflurane and group D (n = 13) received desflurane. Parameters recorded included cardiopulmonary data, body temperature, end‐tidal anaesthetic concentration, duration of anaesthesia, and recovery times and quality. Qualitative data were compared using chi‐squared and Fisher's exact tests and quantitative data with anova and Kruskal–Wallis test. Post‐hoc comparisons for quantitative data were undertaken with the Mann–Whitney U‐test.ResultsThe duration of anaesthesia [mean and standard deviation (SD)] in group I was: 105.3 (27.48) minutes, group S: 120.67 (19.4) minutes, and group D: 113.69 (26.68) minutes (p = 0.32). Times to extubation [group I: 8 minutes, (interquartile range 6–9.5), group S: 7 minutes (IQR 5–7), group D: 5 minutes (IQR 3.5–7), p = 0.017] and to sternal recumbency [group I: 11 minutes (IQR 9.5–13.5), group S: 9.5 minutes (IQR 7.25–11.75), group D: 7 minutes (range 3.5–11.5), p = 0.048] were significantly different, as were times to standing. One dog, following sevoflurane, had an unacceptable quality of recovery, but most other recoveries were calm, with no significant difference between groups.Conclusions and clinical relevanceAll three agents appeared suitable for use. Dogs’ tracheas were extubated and the dogs recovered to sternal recumbency most rapidly after desflurane. This may be advantageous for animals with some neurological diseases and for day case procedures.