Resveratrol oligomers isolated from Carex species inhibit growth of human colon tumorigenic cells mediated by cell cycle arrest

Research has shown that members of the Carex genus produce biologically active stilbenoids including resveratrol oligomers. This is of great interest to the nutraceutical industry given that resveratrol, a constituent of grape and red wine, has attracted immense research attention due to its potential human health benefits. In the current study, five resveratrol oligomers (isolated from Carex folliculata and Carex gynandra ), along with resveratrol, were evaluated for antiproliferative effects against human colon cancer (HCT-116, HT-29, Caco-2) and normal human colon (CCD-18Co) cells. The resveratrol oligomers included one dimer, two trimers, and two tetramers: pallidol (1); α-viniferin (2) and trans-miyabenol C (3); and kobophenols A (4) and B (5), respectively. Although not cytotoxic, the resveratrol oligomers (1-5), as well as resveratrol, inhibited growth of the human colon cancer cells. Among the six stilbenoids, α-viniferin (2) was most active against the colon cancer cells with IC(50) values of 6-32 μM (>2-fold compared to normal colon cells). Moreover, α-viniferin (at 20 μM) did not induce apoptosis but arrested cell cycle (in the S-phase) for the colon cancer but not the normal colon cells. This study adds to the growing body of knowledge supporting the anticancer effects of resveratrol and its oligomers. Furthermore, Carex species should be investigated for their nutraceutical potential given that they produce biologically active stilbenoids such as α-viniferin.     

Radical scavenging capacity and antimutagenic properties of purified proteins from Solanum betaceum fruits and Solanum tuberosum tubers

In this study, antioxidant activities in free-radical-mediated oxidative systems and the genotoxic/antigenotoxic effects of two proteins with molecular mass around 17 kDa, purified from Solanum betaceum fruits (cyphomine) and Solanum tuberosum tubers (solamarine), were investigated. Both proteins inhibited uric acid formation with IC(50) values between 55 and 60 μg/mL, and both proteins were able to reduce oxidative damage by scavenging hydroxyl radicals and superoxide anion in a dose-dependent manner. Furthermore, the DPPH? reduction assay showed SC(50) values of 55-73 μg/mL. Cyphomine and solamarine were able to retain their antioxidant activity after heat treatment at 80 °C for 15 min. Allium cepa and Salmonella /microsome assays showed no genotoxic and mutagenic effects. Solamarine showed an antimutagenic effect against a direct mutagen (4-nitro-o-phenylenediamine). Consequently, the present study showed that the investigated proteins are promising ingredients for the development of functional foods with a beneficial impact on human health and an important source for the production of bioactive peptides.     

Biochemical properties of bioplastics made from wheat gliadins cross-linked with cinnamaldehyde

The aim of this work has been to study the modification of gliadin films with cinnamaldehyde as a potential cross-linker agent. The molecular weight profile and cross-linking density showed that cinnamaldehyde increased reticulation in the resulting films. The participation of free amino groups of the protein in the newly created entanglements could be a possible mechanism of connection between the polypeptidic chains. The combination of a Schiff base and a Michael addition is a feasible approach to understanding this mechanism. The protein solubility in different media pointed to lower participation by both noncovalent and disulfide bonds in stabilizing the structure of the cross-linked films. The new covalent bonds formed by the cinnamaldehyde treatment hampered water absorption and weight loss, leading to more water-resistant matrices which had not disintegrated after 5 months. The properties of this novel bioplastic could be modified to suit the intended application by using cinnamaldehyde, a naturally occurring compound.     

Hepatic immune response in calves during acute subclinical infection with bovine viral diarrhoea virus type 1

Eight colostrum-deprived calves aged 8-12 weeks were inoculated intranasally with a non-cytopathic strain of bovine viral diarrhoea virus (BVDV) genotype-1 and the effects on the hepatic immune response were studied. Two calves were sacrificed at each of 3, 6, 9 and 14 days post-inoculation (dpi) and two uninoculated animals were used as negative controls. BVDV was detected in hepatic macrophages and monocytes from 3 to 14dpi and in Küpffer cells (KCs) from 6 to 14dpi. Increases in the numbers of MAC387(+) KCs and monocytes, but not interstitial macrophages, differentiated by morphological features, were evident in the liver following inoculation with BVDV. There was a substantial increase in the number of monocytes positive for tumour necrosis factor (TNF)-α, but only small increases in the numbers of TNF-α(+) KCs and interstitial macrophages and interleukin (IL)-6(+) monocytes, KCs and interstitial macrophages. There was an increase in the number of interstitial CD3(+) T lymphocytes in the liver, but no substantial changes in the numbers of circulating CD3(+) T lymphocytes, interstitial or circulating CD4(+) or CD8(+) T lymphocytes, or CD79αcy(+) B lymphocytes. Serum haptoglobin and serum amyloid A increased transiently at 12dpi. Upregulation of some pro-inflammatory cytokines by hepatic macrophages is evident in subclinical acute BVDV type 1 infection in calves.     

Using BCG,MPT-51 and Ag85 as antigens in an indirect ELISA for the diagnosis of bovine tuberculosis

This study evaluated the Mycobacterium tuberculosis protein antigen MPT-51, the trimeric antigen 85 (Ag85) complex, and Bacillus Calmette-Guérin (BCG) in an indirect ELISA to diagnose bovine tuberculosis (TB) from serum samples. Serum was collected from 208 intra-dermal tuberculin test (ITT)-positive and 54 ITT-negative animals from a region where bovine TB is endemic. Using the Ag85 and BCG antigens, the indirect ELISA was able to discriminate ITT-positive from ITT-negative animals. This level of discrimination was not achieved when using the MPT-51 antigen. The highest sensitivity (Se) and specificity (Sp) of the test was found when BCG was used (Se, 82%; Sp, 91%). Further work in different epidemiological settings and with larger numbers of animals will be required to validate these findings.     

Transmission electron microscopy for characterization of acrosomal damage after Percoll gradient centrifugation of cryopreserved bovine spermatozoa

The objective of this study was to characterize acrosomal ultrastructure following discontinuous Percoll gradient centrifugation of cryopreserved bovine sperm. Semen was collected from six bulls of different breeds and three ejaculates per bull were evaluated. Frozen semen samples were thawed and the acrosomal region of sperm cells was evaluated by transmission electron microscopy (TEM) before (n = 18) and after (n = 18) Percoll centrifugation. The evaluation of 20 sperm heads from each of the 36 samples analyzed ensured that a large number of cells were investigated. The data were subjected to analysis of variance at a level of significance of 5%. Percoll centrifugation reduced the percentage of sperm exhibiting normal acrosomes (from 61.77 to 30.24%), reduced the percentage of sperm presenting atypical acrosome reactions (from 28.38 to 4.84%) and increased the percentage of sperm exhibiting damage in the acrosome (from 6.14 to 64.26%). The percentage of sperm with typical acrosome reactions was not significantly different before (3.70%) and after (0.67%) centrifugation. TEM distinguished four different types of acrosomal status and enabled ultrastructural characterization of acrosomal injuries. The percentage of sperm exhibiting normal acrosomes decreased and damage in the acrosome was the most frequent acrosomal injury with the Percoll gradient centrifugation protocol utilized.     

Comparison of a rebound and an applanation tonometer for measuring intraocular pressure in normal rabbits

Objective To compare intraocular pressure (IOP) measurements made on healthy adult rabbits without the effect of tranquilizers using the new applanation tonometer, Tono‐Pen Avia^®, and the rebound tonometer Tonovet^®. Methods Intraocular pressure was measured throughout the day (6:00, 9:00, 12:00, 15:00, and 18:00 h) in 38 adult New Zealand White rabbits (76 eyes). The animals were 20 males and 18 females, with a mean weight of 3.5 kg and an average age of 6 months. A complete ocular exam (including Schirmer tear test, fluorescein staining, slit‐lamp biomicroscopy, and direct ophthalmoscopy) was performed on all animals at the beginning of the trial. Rebound tonometry was performed, and after 10 min, anesthetic drops were instilled and applanation tonometry was carried out. IOP values obtained using the two techniques were analyzed statistically. Results The mean IOP was 9.51 ± 2.62 mmHg with Tonovet^®, and 15.44 ± 2.16 mmHg with the Tono‐Pen Avia^®. Significant differences between measurements with the two tonometers were observed (P < 0.001). The linear regression equation describing the relationship between the two tonometers was y = 0.4923x + 10.754 (y = Tonovet^® and x = Tono‐Pen Avia^®). High IOPs were recorded in the early measurements (6:00), but the average IOPs from both devices were statistically similar throughout the day (P = 0.086). The correlation coefficient was r² = 0.357. No significant difference in IOP regarding gender was observed. Conclusion The Tono‐Pen Avia^® recorded higher levels of IOP compared with the Tonovet^®. Early in the day, the IOP of rabbits was higher than later in the day, regardless of the tonometer used.     

The effect of diets supplemented with fish broth and fish oil on the health of weaners

The aim of this study was to determine the effect of fish-based feed materials, as a source of readily available protein contained in fish broth and essential polyunsaturated fatty acids (PUFAs) found in fish oil, on the health of piglets and rearing results. The experiment was conducted on a commercial pig fattening farm. The study involved a total of 80 weaners with an approximate body weight of 15 kg. The experiment was carried out over a period of 40 days. Feed samples were subjected to laboratory analyses. Blood samples were collected from experimental group animals to determine serum biochemical and immunological parameters. The body weight gains of weaners, mortality rates and average feed intake per animal were calculated for the entire experimental period. The addition of fish broth and fish oil significantly improved the n3:n6 fatty acid ratio in diets. The presence of EPA and DHA in the experimental diet could have had a positive health effect on piglets, comparable with that exerted by therapeutic doses of zinc often administered to pigs of this age group. During the experiment, feed conversion ratio (FCR) gain was considerably reduced in the experimental group, with similar daily gains in the control and experimental group.     

Prevalence of Salmonella and Yersinia in free-living pied flycatchers (Ficedula hypoleuca) in central Spain

Salmonella and Yersinia are important enteropathogens in poultry and can affect birds of all ages, including embryos. These food-borne zoonotic enteropathogens are of great economic and medical concern worldwide and are intensely studied in poultry. Information regarding the prevalence of these bacteria in wild birds is scarce and biased toward avian species ecologically linked to humans, which have often been incriminated as both reservoirs and disseminators of these enteropathogens. The prevalence of Salmonella and Yersinia recovered from both the feces and eggs in a population of female pied flycatchers (Ficedula hypoleuca) breeding in nest-boxes in central Spain was evaluated. Salmonella arizonae was recovered from the feces of one female but was not recovered from eggs. Yersinia was not detected in either the feces or eggs. These results may suggest that Salmonella and Yersinia may be uncommon in this population studied and may indicate that these birds are unlikely reservoirs of Salmonella and Yersinia.     

Expression of 14-3-3 σ protein in normal and neoplastic canine mammary gland

14-3-3 σ protein is a negative cell cycle regulator, with both reduced and elevated levels associated with cancer in humans. This study assessed the expression of this protein in canine mammary tissues using immunohistochemistry and Western blotting. 14-3-3 σ was detected in 97% of the mammary tissue samples examined and was found in both myoepithelial (MECs) and epithelial (ECs) cells. Expression levels were elevated and reduced in neoplastic ECs and MECs, respectively (P < 0.001). Intense expression of 14-3-3 σ was detected in neoplastic ECs infiltrating blood vessels and lymph nodes and suggests a possible role for this protein in the malignant transformation of mammary neoplasms. Moreover, double immunostaining for 14-3-3 σ and the MEC – specific marker p63, confirmed that 14-3-3 σ is a highly sensitive marker of MECs since all p63 – positive cells were also positive for 14-3-3 σ. However, this protein is not exclusive to MECs as ECs also labelled positively.