Identification of beer bitter acids regulating mechanisms of gastric acid secretion

Beer, one of the most consumed beverages worldwide, has been shown to stimulate gastric acid secretion. Although organic acids, formed by fermentation of glucose, are known to be stimulants of gastric acid secretion, very little is known about the effects of different types of beer or the active constituents thereof. In the present study, we compared the effects of different beers on mechanisms of gastric acid secretion. To investigate compound-specific effects on mechanisms of gastric acid secretion, organic acids and bitter compounds were quantified by HPLC-DAD and UPLC-MS/MS and tested in human gastric cancer cells (HGT-1) by means of a pH-sensitive fluorescent dye which determines the intracellular pH as an indicator of proton secretion. The expression of relevant genes, coding the H(+)/K(+)-ATPase, ATP4A, the histamine receptor, HRH2, the acetylcholine receptor, CHRM3, and the somatostatin receptor, SSTR2, was determined by qPCR. Ethanol and the organic acids succinic acid, malic acid, and citric acid were demonstrated to contribute to some extent to the effect of beer. The bitter acids comprising α-, β-, and iso-α-acids were identified as potential key components promoting gastric acid secretion and up-regulation of CHRM3 gene expression by a maximum factor of 2.01 compared to that of untreated control cells with a correlation to their respective bitterness.     

Dysphagia caused by focal guttural pouch mycosis: mononeuropathy of the pharyngeal ramus of the vagal nerve in a 20-year-old pony mare

A 20-year-old pony mare was presented to the equine hospital with a ten-day history of dysphagia, regurgitation and coughing. An obstruction of the oesophagus was excluded via endoscopy, but the proximal oesophagus appeared to be distended and circular contractions were missing. A guttural pouch endoscopy revealed a single, black-mottled plaque on the pharyngeal ramus of the vagus nerve in the left guttural pouch, causing a local swelling of this nerve. The pharyngeal ramus seemed to be atrophic distal to the lesion. A biopsy was taken from the lesion and histopathological findings proved the reasonable suspicion of a guttural pouch mycosis with a high degree of purulent-necrotic inflammation and invasion of fungal hyphae. There were no signs of neoplasia, such as melanoma. Daily guttural pouch irrigations with a clotrimazole emulsion (20 g Canesten^® Gyn⁴ solved in 500 ml water), led to a good recovery of the mucosa above the nerve. Periodic endoscopic examination of the left guttural pouch showed that local thickening and distal atrophy of this pharyngeal ramus did not improve, neither did the clinical symptoms. Due to progressive weight loss, acute respiratory distress and aspiration pneumonia, the 20-year-old pony mare unfortunately had to be euthanized three weeks after discharge. This case report emphasizes the enormous importance of a single nerve for the realization of the swallowing process. The one-sided loss of function of the pharyngeal branch of the vagal nerve cannot be compensated neither by the remaining ipsilateral nerves nor by the contralateral normal functioning glossopharyngeal and vagal nerves and thus inevitably leads to severe dysphagia.     

Incidence of Insect Bite Hypersensitivity in a Small Population of Warmblood Horse Breed in the Czech Republic

Insect bite hypersensitivity (IBH) is a serious equine disease with a complex etiology, affecting many breeds. The availability of a small closed population with a relatively high incidence of IBH (28%) offered the chance to contribute to a better understanding of the etiology of IBH. All 736 Warmblood horses, kept in lowland floodplain meadows in the Czech Republic, were analyzed in the period 1998-2010. The morbidity rate in the offsprings of particular stallions was significantly different; the lowest was 10% and the highest 75%. The assumption that the highest occurrence of healthy offspring would be in healthy parents and vice versa was confirmed. The sex of the offsprings was not significant. To decline the disorder prevalence in this population, stud stallions should be IBH negative; afflicted mares can be tolerated to a certain extent, but their male offspring must never be used as stud. Heredity control is necessary, and stallions with IBH+ offspring should be culled. There is to date no reliable treatment available, and therefore, the development of preventive measures is particularly important. The incidence of IBH could be lowered by prudent breeding, and thus immunotherapy and complex herd management would not be necessary. However, this would be a long-term and complex process.     

Epidural myelolipoma in a Husky-cross: a case report

Epidural spinal myelolipoma was diagnosed in an 11.5-year-old castrated male Husky-cross that was evaluated at the veterinary teaching hospital due to progressive thoracolumbar spinal hyperaesthesia and mild proprioceptive pelvic limb ataxia. A focal, ill-defined mildly inhomogenous extradural mass lesion was detected by MRI. The dog was euthanized. At necropsy an extradurally located reddish mass of about 2.5 cm in diameter was present in the vertebral canal. The mass was identified histopathologically as an epidural myelolipoma.     

A finite element model of the bovine claw under static load for evaluation of different flooring conditions

AIM: To create a computer-based finite element (FE) model of the bovine claw and to use finite element analysis (FEA) to estimate stress and deformation of a physiologically-shaped model claw under static load, to visualise potential material weakness and to evaluate the effect of different flooring conditions.

METHODS: Model geometry was derived using digitalised images from a recently trimmed, sound, hind claw from a 4-year-old Austrian Fleckvieh cow. Material properties of bovine claw horn were defined from preliminary investigations and recently established material data, using a modulus of elasticity from 200 to 600 N/mm² . Meshing of the model was performed with 42,127 elements based on 116,141 nodes. Loading of the model was defined at 756 N per claw on a hard and soft surface.

RESULTS: The FE model of the bovine claw under a load of 756 N showed only minimal deformation, most of which took place at the axial wall. Highest stresses were evident in the proximal axial wall, the outer edge of the weight-bearing surface and under the heels. The claw-floor contact image showed a pressure distribution resembling the distal rim of the claw wall. On the hard surface, the maximum stresses were three times higher than those on the soft surface.

CONCLUSIONS AND CLINICAL RELEVANCE: FEA allowed visualisation of the effects that loading on different floor surfaces have on the biomechanics of the claw. Uneven preparation of the claw sole resulted in high stresses at and close to irregularities of the sole. Consequences were more severe on harder flooring. The model supports the hypothesis that mechanical factors play a substantial role in the pathogenesis of claw lesions.     

Concentration of extracellular vesicles isolated from blood relative to the clinical pathological status of dogs with mast cell tumours

Extracellular vesicles (EVs) are membrane‐enclosed fragments shed from all cell types, including tumour cells. EVs contain a wide range of proteins, biolipids and genetic material derived from mother cells and therefore may be potential biomarkers for tumour diagnosis, disease progression and treatment success. We studied the effect of canine mast cell tumours (MCTs) on EV concentrations in blood isolates in association with MCT's histological grade, Ki‐67 proliferative index, KIT‐staining pattern and number of PLT. The average EV concentration in blood isolates from nine dogs with MCTs was considerably higher than that in blood from eight healthy dogs. But there were no statistically significant differences in EVs concentration in the population of dogs with MCT according to a different histological grade of malignancy (Patnaik, Kiupel), KIT‐staining pattern and Ki‐67 proliferation index. The results show that these variables statistically do not significantly predicted EV concentrations in blood isolates (P > .05), except the KIT‐staining pattern I which added statistically significantly to the prediction (P < .05). The results confirmed the impact of neoplasms on the morphological changes to cell membranes, which result in greater vesiculability and higher EV concentrations.     

Modelling the effects of forest management intensification on base cation concentrations in soil water and on tree growth in spruce forests in Sweden

The study investigated the effects of forest residue extraction on tree growth and base cations concentrations in soil water under different climatic conditions in Sweden. For this purpose, the dynamic model ForSAFE was used to compare the effects of whole-tree harvesting and stem harvesting on tree biomass and the soil solution over time at 6 different forest sites. The study confirmed the results from experimental sites showing a temporary reduction of base cation concentration in the soil solution for a period of 20–30 years after whole-tree harvesting. The model showed that this was mainly caused by the reduced inputs of organic material after residue extraction and thereby reduced nutrient mineralisation in the soil. The model results also showed that whole-tree harvesting can affect tree growth at nitrogen-poor forest sites, such as the ones in northern Sweden, due to the decrease of nitrogen availability after residue removal. Possible ways of reducing this impact could be to compensate the losses with fertilisation or extract residue without foliage in areas of Sweden with low nitrogen deposition. The study highlighted the need to better understand the medium- and long-term effects of whole-tree harvesting on tree growth, since the results suggested that reduced tree growth after whole-tree harvesting could be only temporary. However, these results do not account for prolonged extraction of forest residues that could progressively deplete nutrient pools and lead to permanent effects on tree growth.

     

Biochar-application rate and method affect nutrient availability and retention in a coarse-textured,temperate agricultural Cambisol in a microcosm experiment

Background

Agricultural soils often require organic amendments, which improve crop yield and ecosystem services. Biochar has been proven to increase nutrient availability and retention in fine-textured, tropical soils.

Aims

Here we determine how coarse-textured, temperate soils react to different biochar-application rates in different tillage systems.

Methods

We conducted a 6-month laboratory incubation experiment in microcosms filled with a coarse-textured, temperate agricultural soil to determine the effects of biochar-application rate (none, low, or high, i.e., 0, 20, or 40 t dw ha⁻¹, respectively) and application method (mixed into the soil or applied to the soil surface) on microbial activity and biomass, and nutrient availability and leaching.

Results

Microbial activity and biomass and contents of carbon, nitrogen, and phosphorus in leachates were higher in biochar-addition treatments (by 134%, 37%, 372%, 28%, and 801%, respectively) than in the no-addition treatment. The effect was stronger with the low than with the high biochar-application rate. Biochar applied by both methods acted as a slow-release fertilizer, but this effect was stronger when biochar was mixed into the soil. Although available nutrient contents in the soil remained high, nutrient leaching decreased with incubation time. This effect was especially evident when biochar was mixed into the soil.

Conclusions

Biochar is an effective organic amendment in coarse-textured soils providing available nutrients. On the other hand, nutrient-retention mechanisms develop slowly after biochar application and may be greater when biochar is mixed into the soil than applied on the soil surface.     

The inability of tapeworm Hymenolepis diminuta and fluke Dicrocoelium dendriticum to metabolize praziquantel

Biotransformation enzymes can, to a certain extent, protect parasitic worms against the toxic effects of anthelmintics and can contribute to drug-resistance development. The objective of our work was (1) to find and identify phase I and II metabolites of the anthelmintic praziquantel (PZQ) formed by the lancet fluke (Dicrocoelium dendriticum) and the rat tapeworm (Hymenolepis diminuta) and (2) to compare PZQ metabolites in helminths with PZQ biotransformation in rat as host species. Ultra high performance liquid chromatography/tandem mass spectrometry (UHPLC/MS/MS) was used for this purpose. During in vitro incubations, mitochondria-like and microsomes-like fractions (prepared from homogenates of adult worms or from rat liver homogenate) were incubated with 10 and 100 μM PZQ. Liquid/liquid extraction was used for samples during in vitro experiments. In the ex vivo study, living D. dendriticum and H. diminuta adults were incubated in RPMI-1640 medium in the presence of 50 nM or 100 nM PZQ for 24h. After incubation, the worms were removed from the medium and homogenized. Homogenates of worms, medium from the incubation of worms or rat hepatocytes and rat urine (collected during 24h after oral PZQ administration) were separately extracted using solid-phase extraction. The results showed that both D. dendriticum and H. diminuta enzymatic systems are not able to metabolize PZQ. On the other hand, thirty one different phase I and four phase II PZQ metabolites were detected in rat samples using UHPLC/MS/MS analyses. These results show that our experimental helminths, as the members of tapeworm and fluke groups of parasites, are not able to deactivate PZQ, and that the biotransformation enzymes of the studied helminths do not contribute to PZQ-resistance.     

Effect of sainfoin (Onobrychis viciifolia) silage and hay on established populations of Haemonchus contortus and Cooperia curticei in lambs

The objective of the study was to examine the effect of dried and ensiled sainfoin (Onobrychis viciifolia) on established populations of Haemonchus contortus (abomasum) and Cooperia curticei (small intestine) in lambs under controlled conditions. Twenty-four parasite na?ve lambs were inoculated with a single dose of infective larvae of these parasites 28 days prior to the start of the feeding experiment. Twenty-four days post-infection, 4 days prior to the start of the feeding experiment, animals were allocated to four groups according to egg excretion, live weight and sex. Groups A and B received sainfoin hay and control hay, respectively, for 16 days. Groups C and D were fed on sainfoin silage or control silage for the same period. Feeds were offered ad libitum and on the basis of daily refusals were supplemented with concentrate in order to make them isoproteic and isoenergetic. Individual faecal egg counts on a dry matter basis (FECDM) were performed every 3-4 days and faecal cultures and packed cell volume (PCV) measurements were done weekly. After 16 days of experimental feeding, all animals were slaughtered and adult worm populations were determined. The consumption of conserved sainfoin was associated with a reduction of adult H. contortus (47% in the case of hay, P<0.05; 49% in the case of silage, P=0.075) but had little effect on adult C. curticei. Compared to the controls, H. contortus specific FECDM was reduced by 58% (P<0.01) in the sainfoin hay group and by 48% (P=0.075) in the sainfoin silage group. For both sainfoin feeds FECDM specific to C. curticei were significantly decreased when compared to the control feeds (hay 81% and silage 74%, both tests P<0.001). Our data suggest that different mechanisms were responsible for the reduction in FECDM in response to feeding tanniferous fodder. For H. contortus, the decrease seemed to be due to a nematocidal effect towards adult H. contortus. In contrast for C. curticei, the reduction in FECDM appeared to be a result of a reduced per capita fecundity. For both, hay and silage, an antiparasitic effect could be shown, offering promising perspectives for the use of conserved tanniferous fodder as a complementary control approach against GIN.