Identification of Cytokeratins in Bovine Sperm Outer Dense Fibre Fractions

Outer dense fibres (ODF) are important substructures of mammalian sperm tails that are involved in the regulation of sperm motility. In this study, we investigated the identity of several sodium dodecyl sulphate (SDS)‐insoluble ODF proteins. Bovine ODF were purified by separating sperm heads and tails using ultrasound and Percoll^® density gradient centrifugation. Sperm flagella were treated with the detergent cetyltrimethylammonium bromide (CTAB). CTAB‐insoluble material, which reportedly represents the ODF fraction, was collected, and electron microscopy confirmed a highly purified ODF fraction. We found after solubilization of this fraction with SDS that high amounts of insoluble material were retained after centrifugation. SDS‐insoluble material was collected and quantitatively dissolved in 8 M urea. SDS‐gel electrophoresis in the presence of urea revealed polypeptides with apparent molecular masses of approximately 25, 43, and 50 kDa. Subsequent immunoblotting with anti‐cytokeratin antibodies detected two urea‐soluble, SDS‐insoluble proteins with apparent molecular masses of 45 and 66 kDa. The 45‐kDa protein was identified as cytokeratin 19. An antibody reacting with a palette of cytokeratins (CK 1–18 and CK 20), KL1, was the only antibody that reacted with the 66‐kDa polypeptide. We conclude that sperm ODF fractions contain at least one each of type I and type II intermediate filaments. As keratins and intermediate filaments are described as rope‐like structures, we suggest that these intermediate filaments play an important structural or tension‐bearing role in sperm flagella.     

Diversity in tropical rain forests and coral reefs

The commonly observed high diversity of trees in tropical rain forests and corals on tropical reefs is a nonequilibrium state which, if not disturbed further, will progress toward a low-diversity equilibrium community. This may not happen if gradual changes in climate favor different species. If equilibrium is reached, a lesser degree of diversity may be sustained by niche diversification or by a compensatory mortality that favors inferior competitors. However, tropical forests and reefs are subject to severe disturbances often enough that equilibrium may never be attained.     

Effects of Confluent, Roscovitine Treatment and Serum Starvation on the Cell-cycle Synchronization of Bovine Foetal Fibroblasts

The present study was designed to examine the effects of cell-cycle synchronization protocols, such as confluent, roscovitine treatment and serum starvation, in bovine foetal fibroblasts on synchronization accuracy at G0/G1, viability, apoptosis, necrosis and ploidy for use as a nuclei donor. The cells in 5-10 passages were randomly allocated into three treated groups. Cells were cultured either in Dulbecco's modified Eagle's medium (DMEM) + 10% foetal bovine serum (FBS) until 90% confluent (group 1, confluent), in DMEM + 10% FBS + 30 microM roscovitine for 12 h (group 2, roscovitine), or in DMEM + 0.5% FBS for 5 days (group 3, serum starvation). Most of the cells (>80%) in all groups were arrested at the G0/G1 stage. Although the rates did not differ, cells in group 1 showed an increased cell population arrested at the G0/G1 phase. Significantly (p < 0.05) higher rates of apoptosis occurred in group 3 than in group 1 and 2 (10% vs 6% and 6%, respectively). No differences in chromosomal abnormality were observed among groups. However, by increasing the number of cell culture passages up to 15, significantly (p < 0.05) higher chromosomal abnormality was observed than in 5 and 10 passages (39% vs 28% and 23%, respectively) in group 1. The results clearly indicated that bovine foetal fibroblasts could be effectively synchronized at G0/G1 stages by all the three different treatments, confluent, roscovitine and serum starvation. However, cells in confluent showed reduced apoptosis and necrosis when they underwent 5-10 passages, exhibiting increased percentage of cells with stable chromosome diversity. Hence, cells in confluent merit further studies before they could be used as nuclear donors.     

Successful small intestinal resection and anastomosis in a late term broodmare with colic via a standing left flank laparotomy

A late term broodmare presented with low‐grade intestinal colic. Clinical findings were consistent with a small intestinal lesion requiring surgical intervention. The risks of general anaesthesia to the fetus, combined with clinical findings in an otherwise quiet natured horse influenced the decision to choose an alternative standing left flank approach first. A small intestinal resection and anastomosis was performed successfully and the mare delivered a healthy foal without complications a month later. Standing colic surgery might be a preferred alternative to general anaesthesia in certain circumstances as discussed in this case report.     

Theileria orientalis: a review

Theileria orientalis (also known historically as T. sergenti and T. buffeli) is responsible for benign or non-transforming theileriosis, and exerts its major effect through erythrocyte destruction. The life cycle of T. orientalis is essentially similar to that of other Theileria species, except that the schizonts do not induce transformation and fatal lymphoproliferation. The pathogenesis of anaemia as a result of infection is not clearly established and may be multifaceted. Clinical signs of weakness, reluctance to walk and abortion are early but non-specific indications of disease, particularly if accompanied by a history of cattle being moved. Physical examination may reveal pallor (pale eyes, vaginal mucosa), pyrexia, and elevated heart and respiratory rates. T. orientalis is an economically important parasite of cattle in New Zealand, Australia and Japan, especially where naïve animals are introduced into an endemic area or in animals under stress. Increased awareness of the risks posed by the parasite is required to enable management practices to be implemented to minimise its impact.     

The potential of mini-ridging for controlling intrarow weeds: estimating minimum lethal burial depth

Weed management using synthetic herbicides is undergoing a global decline, necessitating a re-evaluation of existing control measures and the development of novel weed management tools. ‘Mini-ridging’ is a non-discriminatory, physical weeding method that functions by burying weeds in the intrarow with a laterally shifted ridge of soil. In glasshouse trials using potted plants, we found that plant recovery after soil application was influenced by plant size, which in turn was influenced by plant species, developmental stage and/or age. The likelihood of plant recovery after soil application was negatively related to the depth of soil applied: very few plants survived total coverage by soil but, conversely, survival could be substantial if some parts of the plants were not covered. The results suggest that burial under a depth of 6 cm of soil would eliminate most plants regardless of species or growth stage. Larger plants would require the application of a greater total depth of soil to achieve this 6 cm of soil cover, and weed management would, therefore, tend to be more successful and more practical if weeds were targeted when still small. This research demonstrates the potential of plant burial as a simple and reliable means of non-chemical weed management, and re-emphasises that, for weed control to be effective, the applied soil layer must cover the whole plant.     

The sounds of silence: regime shifts impoverish marine soundscapes

Context

Regime shifts are well known for driving penetrating ecological change, yet we do not recognise the consequences of these shifts much beyond species diversity and productivity. Sound represents a multidimensional space that carries decision-making information needed for some dispersing species to locate resources and evaluate their quantity and quality.

Objectives

Here we assessed the effect of regime shifts on marine soundscapes, which we propose has the potential function of strengthening the positive or negative feedbacks that mediate ecosystem shifts.

Methods

We tested whether biologically relevant cues are altered by regime shifts in kelp forests and seagrass systems and how specific such shifted soundscapes are to the type of driver; i.e. local pollution (eutrophication) vs. global change (ocean acidification).

Results

Here, we not only provide the first evidence for regime-shifted soundscapes, but also reveal that the modified cues of shifted ecosystems are similar regardless of spatial scale and type of environmental driver. Importantly, biological sounds can act as functional cues for orientation by dispersing larvae, and observed shifts in soundscape loudness may alter this function.

Conclusions

These results open the question as to whether shifted soundscapes provide a functional role in mediating the positive or negative feedbacks that govern the arrival of species associated with driving change or stasis in ecosystem state.

     

含莫纳可林K红曲降低鸡蛋胆固醇效果的研究

48只68周龄的海兰褐蛋鸡分成三组，按0.5％和10％比例分别添加含0．4％莫纳可林K红曲于各组日粮内，饲喂22d，观测其对鸡蛋胆固醇变化的影响。结果表明，投药后5—6d，10％添加组和5％添加组蛋黄胆固醇开始下降；15—22d的胆固醇含量与1～5d相比，10％添加组和5％添加组分别下降了9．02％和5．10％，而对照组升高了5．22％。     

The spore coat of the bean anthracnose fungus Colletotrichum lindemuthianum is required for adhesion, appressorium development and pathogenicity

The spores (conidia) of the bean anthracnose fungal pathogen, Colletotrichum lindemuthianum, adhere to the aerial parts of plants to initiate the infection process. In previous studies we have shown that the Colletotrichum spores are surrounded by a fibrillar spore coat, comprising several major glycoproteins. Previous evidence showed that a monoclonal antibody (UB20) that recognised these glycoproteins was able to inhibit adhesion of spores to a hydrophobic surface. In this paper we have further studied the role of the spore coat in adhesion, germination and fungal development by studying the effects of UB20 and protease treatment of spores. The latter treatment has previously been shown to remove the spore coat. Spores germinate on glass, polystyrene and water agar, however, appressoria only develop on glass or polystyrene, showing a requirement for a hard surface. Removal of the spore coat with protease inhibits adhesion at 30 min, before the secretion of ECM glycoproteins. Protease treatment also inhibits the development of appressoria and reduces pathogenicity on leaves. Incubation of spores with the MAb UB20 inhibits adhesion at 30 min, but does not affect appressorium formation or pathogenicity. The results suggest that an intact spore coat has two functions; it is required for adhesion to a hydrophobic surface and for the detection of a hard surface necessary for appressorium formation. We suggest that contact with a hard surface, rather than adhesion, is the key event leading to appressorium formation.