Apple juice clarification by immobilized pectolytic enzymes in packed or fluidized bed reactors

The catalytic behavior of a mixture of pectic enzymes, covalently immobilized on different supports (glass microspheres, nylon 6/6 pellets, and PAN beads), was analyzed with a pectin aqueous solution that simulates apple juice. The following parameters were investigated: the rate constant at which pectin hydrolysis is conducted, the time (tau(50)) in which the reduction of 50% of the initial viscosity is reached, and the time (tau(comp,dep)) required to obtain complete depectinization. The best catalytic system was proven to be PAN beads, and their pH and temperature behavior were determined. The yields of two bed reactors, packed or fluidized, using the catalytic PAN beads, were compared to the circulation flow rate of real apple juice. The experimental conditions were as follows: pH 4.0, T = 50 degrees C, and beads volume = 20 cm(3). The initial pectin concentration was the one that was present in our apple juice sample. No differences were observed at low circulation rates, while at higher recirculation rates, the time required to obtain complete pectin hydrolysis into the fluidized reactor was found to be 0.25 times smaller than in the packed bed reactor: 131 min for the packed reactors and 41 min for the fluidized reactors.     

A Neolithic yew bow in the Polish Carpathians: Evidence of the impact of human activity on mountainous palaeoenvironment from the Kamiennik landslide peat bog

A unique artefact — the fragment of a bow made of yew wood, indicating the hunting activity of prehistoric man — was found within the sediments of the landslide peat bog (at the depth of 330 cm) formed on the Mt. Kamiennik (Polish Flysch Carpathians). The datings of this artefact using ¹⁴C method at ca 3790–3380 cal BC indicate its connection with the activity of the Neolithic man, related to the Funnel Beaker Culture. In the sequence of the peat bog deposits formed since the Atlantic Phase, the palaeoclimatic changes of the Middle- and Late Holocene, as well as traces of human impact in these and subsequent periods were recorded and analysed using palynological, macrofossils, lithological methods, and dated with radiocarbon. Palaeoenvironmental changes identified in the peat horizon bearing the artefact were marked by the delivery of minerogenic, “high energy” deposits (with charcoal) to the peat bog, as well as distinct changes of plant communities. These phenomena confirm a significant human impact caused by the Prehistoric man on landslide areas. On the other hand, the study confirms significant importance of climatic condition for human activity and proves that landslide peat bog deposits are very sensitive records of palaeoenvironmental changes. The archaeological artefact essentially supplements the results of our interdisciplinary palaeoenvironmental study. It is one of the rare Neolithic bows found in the area of Europe, till now.     

Isolation, identification, and quantification of roasted coffee antibacterial compounds

Coffee brew is a widely consumed beverage with multiple biological activities due both to naturally occurring components and to the hundreds of chemicals that are formed during the roasting process. Roasted coffee extract possesses antibacterial activity against a wide range of microorganisms, including Staphylococcus aureus and Streptococcus mutans, whereas green coffee extract exhibits no such activity. The naturally occurring coffee compounds, such as chlorogenic acids and caffeine, cannot therefore be responsible for the significant antibacterial activity exerted by coffee beverages against both bacteria. The very low minimum inhibitory concentration (MIC) found for standard glyoxal, methylglyoxal, and diacetyl compounds formed during the roasting process points to these alpha-dicarbonyl compounds as the main agents responsible for the antibacterial activity of brewed coffee against Sa. aureus and St. mutans. However, their low concentrations determined in the beverage account for only 50% of its antibacterial activity. The addition of caffeine, which has weak intrinsic antibacterial activity, to a mixture of alpha-dicarbonyl compounds at the concentrations found in coffee demonstrated that caffeine synergistically enhances the antibacterial activity of alpha-dicarbonyl compounds and that glyoxal, methylglyoxal, and diacetyl in the presence of caffeine account for the whole antibacterial activity of roasted coffee.     

Magnetic resonance imaging features of canine incomplete humeral condyle ossification

Incomplete ossification of the humeral condyle (IOHC) is characterized by an intracondylar fissure located where the intercondylar physis is present in growing dogs. Its radiologic and computed tomographic features have been described but the magnetic resonance (MR) features have not been characterized. Our purpose was to further describe the range of MRappearances of IOHC, to assess the diagnostic capability of MRrelative to radiology, and to determine whether MRis able to identify the disease before a fissure forms. Thirty-eight elbow MRscans and radiographs, when available, were reviewed and divided into three groups. In Group 1 (affected elbows, n = 22), there was an intracondylar defect on MRwith variable appearance; the defect was not visible radiographically in 32% of the elbows. The main difference between Group 2 (nonaffected elbows, n = 6) and Group 3 (contralaterals to IOHC or to condylar fracture, without fissure, n = 10) was the appearance of the humeral condyle in short tau inversion recovery (STIR) sequences: all elbows in Group 2 had a homogeneous humeral condyle, whereas all but one in Group 3 were heterogeneous. One dog in Group 3 developed a complete condylar fissure 7 months after the first examination, when no evidence of an intracondylar defect had been detected. The MRappearance of IOHC is variable and a heterogeneous humeral condyle in STIR images without a clear defect may warn of the possibility for the subsequent development of a condylar fissure.     

Anatomical and radiographic study of the scapula in juveniles and adults of Tamandua mexicana (Xenarthra: Myrmecophagidae)

Tamandua mexicana is a mammal of the superorder Xenarthra geographically located between Mexico and Peru that remains insufficiently studied. These species are characterized by their movement, resting, and eating of termites and ants in arboreal and terrestrial environments, and therefore, their thoracic limbs have been adapted morphologically to move in these environments. Tamandua mexicana is a species constantly threatened by different causes, and their thoracic limbs can be affected in the scapular and shoulder regions as a result; therefore, specific anatomical knowledge of the scapula allows for better clinical procedures, surgical approaches, radiological diagnosis and muscular reconstructions in extinct species of anteaters. The main objective of this study was to report the anatomical and radiographic features of the scapula in juvenile and adult specimens of T. mexicana. Gross dissections and radiographic study were performed in six cadavers. The scapula had large differences compared with other mammals, such as two spines, a foramen for the suprascapular nerve, a long acromion, a cranial transverse scapular ligament and caudolateral fossa for the origin of the muscles teres major, tensor fasciae antebrachii and subscapular. In juvenile specimens, a scapular cartilage, an acromial cartilage, craniomedial and caudolateral coracoscapular ligaments forming the foramen for the suprascapular nerve, and progressive ossification of those ligaments and the cranial transverse scapular ligament were observed. These findings were corroborated by radiography; therefore, these characteristics change with age. The bony reliefs of the scapula of T. mexicana corroborate the specialization of the shoulder in this species.     

Effect of caspase inhibitor Z-VAD-FMK on bovine sperm cryotolerance

The aim of this study was to evaluate the treatment of bovine semen with the pan-caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (Z-VAD-FMK), before or after freezing on semen quality. After the initial assessment, sperm from 4 bulls were pooled (Experiment 1) and cryopreserved in BioXcell containing 0, 20 and 100 μM Z-VAD-FMK. After thawing semen viability, motility, membrane integrity, as well as DNA fragmentation and ΔΨm were evaluated. In Experiment 2, bovine frozen/thawed sperm were incubated for 1 hr with 0, 20 and 100 µM Z-VAD-FMK before assessing the semen quality. The treatment with Z -VAD-FMK before cryopreservation improved post-thawing sperm motility compared to the control group (p < .05), while no differences were recorded in sperm viability and membrane integrity among groups (on average 86.8 ± 1.5 and 69.1 ± 1.4, respectively). Interestingly, at the highest concentration, DNA fragmentation decreased (p < .05), while the percentage of spermatozoa with high ΔΨm increased (p < .05). The results of Experiment 2 showed that 1-hr treatment with Z-VAD-FMK did not affect sperm motility and viability (on average 63.4 ± 5.8 and 83.7.1 ± 1.2, respectively). However, Z-VAD-FMK improved sperm membrane integrity (p < .05) and at the highest concentration tested decreased the proportion of sperm showing DNA fragmentation (p < .05). No differences were recorded in the percentage of spermatozoa with high ΔΨm (on average 57.0 ± 11.4). In conclusion, the treatment with 100 µM of the caspase inhibitor Z-VAD-FMK before freezing increased bovine sperm mass motility and ΔΨm, while decreasing sperm DNA fragmentation. Treatment of semen after thawing with 100 µM Z-VAD-FMK improved sperm membrane integrity and reduced DNA fragmentation.     

Virgin olive oil normalizes the altered triacylglycerol molecular species composition of adipose tissue in spontaneously hypertensive rats

The present study was conducted in order to evaluate the influence of hypertension on the triacylglycerol (TG) molecular species composition and other lipid classes of rat adipose tissue. In addition, the effect of two dietary oils, with a similar content in oleic acid but different TG moieties, was studied. Virgin olive oil (VOO) or high-oleic sunflower oil (HOSO) was added to a baseline diet (BD) and administrated to Wistar-Kyoto and spontaneously hypertensive rats (SHR) for 12 weeks. Both VOO and HOSO normalized the altered composition of TG molecular species and phospholipid (PL) fatty acids in SHR compared to animals fed BD, although the effect exhibited by VOO was greater. Rats fed HOSO showed a greater palmitic (p < 0.05) and lower linoleic acid (p < 0.05) incorporation into PL but a greater accumulation of linoleic acid-containing TG species, particularly dioleoyl-linoleoyl-glycerol, with a concomitant displacement of trilinolein. Both oils were capable of increasing the lipoprotein lipase (LPL) activity in normotensive rats, but only VOO did so in the SHR. Therefore, it was concluded that although oleic acid-rich diets improve some of the altered parameters of SHR adipose tissue, VOO is more effective than HOSO in this regard.     

Separation of flavanone-7-O-glycoside diastereomers and analysis in citrus juices by multidimensional liquid chromatography coupled with mass spectrometry

The major flavanone-7-O-glycoside constituents in citrus fruit juices (naringin, hesperidin, neohesperidin, narirutin, and eriocitrin) were separated as diastereomers by multidimensional liquid chromatography. The method consisted of coupling two HPLC columns: a reversed-phase (RP(18)) column was used for the separation of flavanone glycosides, which were, then, individually switched into a carboxymethylated beta-cyclodextrin (beta-CD)-based column and resolved as the corresponding stereoisomers. The method was used for the full analysis of flavanone glycosides in fresh hand-squeezed and commercial fruit juices by combining the quantitative estimation with the diastereomeric analysis. Quantitative data were in general consistent with previously reported data in this field. CC-LC isomer analysis was carried out by coupling the beta-CD column with a mass spectrometer operated with negative ion electrospray ionization (ESI-MS). The results showed that hesperidin was present in orange juices almost exclusively as the 2S isomer, whereas narirutin had mainly the 2R configuration. In grapefruit juices (2S)-naringin prevailed with the respect to the 2R isomer, whereas the opposite was true for narirutin. Lemon juices contained eriocitrin stereoisomers in equal amount (50% each), but hesperidin was almost exclusively found as the 2S isomer. Significant differences of the diastereomeric ratios were observed between freshly squeezed juices and juices from commercial sources.