Nitric oxide impacts bovine sperm capacitation in a cGMP‐dependent and cGMP‐independent manner

We aimed to elucidate whether NO acts in in vitro sperm capacitation in bovine via cGMP/PKG1 pathway. For this, cryopreserved bovine sperm were capacitated in vitro with 20 µg/ml heparin (Control) plus treatments: 1 mM L‐arginine (L‐arg, NO precursor), 50 µM Rp‐8‐Bromo‐β‐phenyl‐1,N²‐ethenoguanosine‐3′,5′‐cyclic monophosphorothioate (Rp‐8‐Br‐cGMPS, selective inhibitor of the binding site for cGMP in PKG1), 1 mM 2‐Phenyl‐4,4,5,5‐tetramethylimidazoline‐1‐oxyl 3‐oxide (PTIO, NO scavenger), and the combinations of L‐arg + RP‐8‐Br‐cGMPS and L‐arg + PTIO. Sperm motility and vigour were determined by phase‐contrast microscopy, capacitation status by chlortetracycline staining, and the intracellular concentration of cGMP was measured by ELISA. Data were subjected to analysis of variance and means compared with SNK test at 5% probability. Motility and vigour were lower in sperm treated with PTIO when compared to Control and other treatments (p < .05). The L‐arg treatment showed the highest percentage of capacitated sperm when compared to the Control and other treatments (Rp‐8‐Br‐cGMPS, L‐arg + Rp‐8‐Br‐cGMPS and PTIO) (69.8 ± 3.4%, 51.2 ± 3.0, 51.1 ± 2.1, 51.2 ± 3.0 and 45.5 ± 2.7, respectively) (p < .05). The capacitation ratio (%) was lower in treatments with Rp‐8‐Br‐cGMPS, L‐arg + Rp‐8‐Br‐cGMPS and PTIO, respectively (p < .05). Lastly, cGMP concentration (pmol/ml) was lower in PTIO and L‐arg + PTIO (1.3 ± 0.3 and 1.6 ± 0.4) and was higher in Rp‐8‐Br‐cGMPS and L‐arg + Rp‐8‐Br‐cGMPS (3.7 ± 0.4 and 4.0 ± 0.5) treatments. We showed that during in vitro capacitation of cattle: (a) NO influences sperm motility and vigour; (b) NO is associated with cGMP synthesis through two independent pathways and (c) the cGMP/PKG1 pathway has a partial role in sperm capacitation and does not involve the L‐arg/NO.     

Evaluation of Particle Size Influence on Proximate Composition,Physicochemical, Techno-Functional and Physio-Functional Properties of Flours Obtained from Persimmon (<Emphasis Type="Italic">Diospyros kaki</Emphasis> Trumb.) Coproducts

The aim of the work was to study the influence of particle size in the composition, physicochemical, techno-functional and physio-functional properties of two flours obtained from persimmon (Diospyros kaki Trumb. cvs. ‘Rojo Brillante’ (RBF) and ‘Triump’ (THF) coproducts. The cultivar (RBF and THF) and particle size significantly affected all parameters under study, although depending on the evaluated property, only one of these effects predominated. Carbohydrates (38.07–46.98 g/100 g) and total dietary fiber (32.07–43.57 g/100 g) were the main components in both flours (RBF and THF). Furthermore, insoluble dietary fiber represented more than 68% of total dietary fiber content. All color properties studied were influenced by cultivar and particle size. For both cultivars, the lower particle size, the higher lightness and hue values. RBF flours showed high values for emulsifying activity (69.33–74.00 mL/mL), while THF presented high values for water holding capacity (WHC: 9.47–12.19 g water/g sample). The bile holding capacity (BHC) and fat/oil binding values were, in general, higher in RBF (19.61–12.19 g bile/g sample and 11.98–9.07, respectively) than THF (16.12–12.40 g bile/g sample and 9.78–7.96, respectively). The effect of particle size was really evident in both WHC and BHC. Due to their dietary fiber content, techno-functional and physio-functional properties, persimmon flours seem to have a good profile to be used as potential functional ingredient.     

Immunohistochemical differentiation of reactive from malignant mesothelium as a diagnostic aid in canine pericardial disease

Objectives

To develop a provisional immunohistochemistry panel for distinguishing reactive pericardium, atypical mesothelial proliferation and mesothelioma in dogs.

Materials and Methods

Archived pericardial biopsies were subject to haematoxylin and eosin staining, immunohistochemistry for cytokeratin, vimentin, insulin‐like growth factor II mRNA‐binding protein 3, glucose transporter 1 and desmin. Samples were scored for intensity and number of cells stained.

Results

Ten biopsies of reactive mesothelium, 17 of atypical mesothelial proliferation, 26 of mesothelioma and five of normal pericardium were identified on the basis of haematoxylin and eosin staining. Cytokeratin and vimentin were expressed in all biopsies, confirming mesothelial origin. Normal pericardial samples had the lowest scores for insulin‐like growth factor II mRNA‐binding protein 3, glucose transporter 1 and desmin. Mesothelioma and atypical proliferative samples were similar to each other, with higher scores for insulin‐like growth factor II mRNA‐binding protein 3 and glucose transporter 1 than the reactive samples. Desmin staining was variable. Insulin‐like growth factor II mRNA‐binding protein 3 was the best to distinguish between disease groups.

Clinical Significance

An immunohistochemistry panel of cytokeratin, vimentin, insulin‐like growth factor II mRNA‐binding protein 3 and glucose transporter 1 could provide superior information compared with haematoxylin and eosin staining alone in the diagnosis of cases of mesothelial proliferation in canine pericardium, but further validation is warranted.     

Driving forces of soil organic matter change in Jiangsu Province of China

Soil organic matter (SOM) is a key property determining soil functions and a major form of carbon stored in soil. Understanding the spatial and temporal variability of SOM and the driving forces responsible for spatial and temporal changes is important to assess regional soil quality and carbon sequestration potential and, particularly, to establish better practices for land use and management. We evaluated the spatio‐temporal change in SOM content from 1979–1982 to 2006 and its driving forces in Jiangsu Province, East China, using geostatistics. The results showed that mean SOM content increased from 16.60 ± 8.50 to 18.31 ± 8.32 g/kg over a 26‐yr period. The maps of SOM generated by ordinary kriging represented the increasing trend from north to south across the province in the two periods. The level of SOM in 1979–1982 affected the pattern of change: the SOM increasing in areas initially with a small content while decreasing in areas having a large content. The map of SOM change showed that the rate of increase decreased from north to south within the province. Increased fertilizer application promoted crop production with more residual biomass being retained in the soil, which resulted in increased SOM content. Land use changes to paddy, upland or forest improved SOM content, whereas abandoning land reduced SOM content.     

Adherence of Actinobacillus pleuropneumoniae to primary cultures of porcine lung epithelial cells

To study adherence of Actinobacillus pleuropneumoniae to porcine lower respiratory epithelium, a cell culture model was developed using primary cultures of porcine lung epithelial cells (LEC). Adherence assays were performed and results were compared with data obtained with swine kidney cells (SK6). A. pleuropneumoniae efficiently adhered to LEC with up to 62 bacteria per cell after 2h of incubation. Reference strain of serotype 3 (R3) adhered better to LEC than reference strains of serotypes 1 (R1), 7 (R7) and 8 (R8). Overall the adherence to LEC was more rapid and up to 30-fold more efficient than adherence to SK6 cells. In search for the mechanism involved in the adherence event, we tested the effect of LPS which has previously been demonstrated to cause adherence of the pathogen to upper respiratory epithelium. Adherence assays with LPS transposon mutants demonstrated unaltered (mutant with modification in core/lipid A moiety) or even three-fold more adherence (mutants lacking O antigen) compared to the parent micro-organisms. Purified LPS of strains R1, R3, R7 and R8 did not inhibit adherence of R8 to LEC either, suggesting that LPS and particularly the O-antigen are not essential for adherence of A. pleuropneumoniae to LEC. The efficient, LPS-independent adherence of A. pleuropneumoniae to LEC cells indicates that A. pleuropneumoniae may carry different, cell type-specific adhesins and that primary cultures of lower respiratory epithelium are valuable infection models in studying A. pleuropneumoniae pathogenesis.     

Redox halopriming: A Promising Strategy for Inducing Salt Tolerance in Bread Wheat

New strategies to enhance growth and productivity of food crops in saline soils represent important research priorities. This study has investigated the role of certain priming techniques to induce salt tolerance of bread wheat. Wheat grains were soaked in 0.2 mm sodium nitroprusside as nitric oxide donor (redox priming), diluted sea water (halopriming) and the combination of both (redox halopriming). Grains were also soaked in distilled water (hydropriming); in addition, untreated grains were taken as control. Our results indicated that priming treatments significantly improved all growth traits and increased leaf pigments concentration as compared to the control. Priming treatments markedly enhanced membrane stability index, proline, total soluble sugars and K⁺ concentration with simultaneous decrease in the concentration of Na⁺ and malondialdehyde (MDA). Furthermore, yield and yield‐related traits such as plant height, spike length, total number of tillers, 1000‐grain weight, straw and grain yield considerably affected by priming treatments. Moreover, the grain yield of both genotypes was positively affected by redox halopriming treatment. However, the extent of enhancement was more prominent in Gemmiza‐9 (salt sensitive) than that in Sakha‐93 (salt‐tolerant). Overall, this study clearly indicated that redox halopriming treatment is a promising and handy technique to induce salinity tolerance of wheat genotypes.     

Unravelling the relationship between a seasonal environment and the dynamics of forage growth in grazed swards

It is well reported in the scientific literature that pastures can have similar net forage accumulation when managed with contrasting structures. However, we hypothesized that the dynamics of forage accumulation in grazed swards is linked to seasonal-environmental conditions. Marandu palisadegrass (Brachiaria brizantha [Hochst. ex A. Rich.] was used as the forage species model. The experimental treatments were four grazing heights (10, 20, 30 and 40 cm) allocated to experimental units according to a randomized complete block design with four replicates and evaluated throughout four contrasting environmental seasons (summer, autumn, winter–early spring and late spring). Under rainy and warm periods, greater net forage accumulation was observed in pastures maintained taller; on the contrary, during the mild and dry periods, net forage accumulation rate reduced as grazing height increased. Such patterns of responses were related to compensations between tiller population density and tissues flows during summer and late spring and the reduced capacity of taller canopies to compensate lower population with greater growth rates during autumn and winter–early spring. Grazed swards changed their patterns of forage growth as they transitioned from favourable to more abiotic stressful conditions, suggesting that seasonal adjustments in grazing intensities are necessary in order to maximize forage production.     

Decolonising Pacific research,building Pacific research communities and developing Pacific research tools: The case of the talanoa and the faafaletui in Samoa

In building Samoan academic researcher capacity in Samoa, we argue that there is a need to first establish the kind of researcher community advocated by Linda Tuhiwai Smith, and to do so through developing research tools, such as the talanoa and faafaletui, in partnership with researcher capacity‐building initiatives such as the applied postgraduate social and health research methods course (coded PUBX731‐HSA505) run by the Centre for International Health, University of Otago, in partnership with the National University of Samoa. This paper offers a commentary on the talanoa and faafaletui as Pacific research methodologies, and asks what its value might be for researchers in Samoa. It reflects on the learning experiences of staff and students of the applied social and health research methods course in relation to the talanoa and faafaletui as Pacific research methodologies or methods. It concludes that developing Pacific research and researcher capacity in Pacific Island countries, such as Samoa, must include opening up spaces within these communities to critically engage what is Pacific or Samoan or indigenous about these research tools, methods or methodologies, and how they might differ in form or substance from other methods or methodologies.