Studies of assisted reproduction in the spotted grass frog Limnodynastes tasmaniensis: ovulation, early development and microinjection (ICSI)

Assisted Reproductive Technologies (ART) offer a wide range of techniques that have the potential to augment efforts to conserve and manage endangered amphibians and improve wild and captive population numbers. Gametes and tissues of species nearing endangered or extinct status can be cryopreserved and stored in gene banks, to provide material that can be utilised in the future as ART methods are refined. The Spotted Grass Frog, Limnodynastes tasmaniensis, is an abundant amphibian species in South-Eastern Australia of the family Myobatrachidae, that is suitable for the development of ART systems that can be applied to the threatened and endangered myobatrachid and other amphibian species native to Australia. The aim of this study was to advance the understanding of ovulation, fertilisation and embryo nic development of Lim. tasmaniensis and in vitro manipulations of reproduction and development for use in the development of advanced ART procedures such as intracytoplasmic spermatozoon injection (ICSI), androgenesis and nuclear transfer. Ovulation in amphibians can be induced by protocols utilising natural or synthetic hormones. All protocols tested on Lim. tasmaniensis in this study required two injections and the most effective protocols continued to require a first injection of pituitary extracts to induce ovulation. The second injection was, however, successfully replaced by synthetic chorionic gonadotrophin at a threshold dosage of 100 iu and halved the number of cane toads required to source the pituitaries. A combination of LHRH and Pimozide offered a less effective protocol, that did not require the use of pituitary extracts, and avoided the risk of pathogen transfer associated with unsterilised pituitary extracts. Unfertilised eggs of Lim. tasmaniensis were exposed to media of various osmolalities to determine media effects on eggs and their surrounding jelly layers that might impact on egg viability and fertilisability. Osmolality had no effect upon the egg diameter, however, rapid swelling of the jelly layers occurred within 15 minutes of exposure to various media treatments and plateaued from 30-90 minutes without further expansion. Swelling of the jelly layers was increased in hypotonic media (2.5% SAR, H2O) and minimised in the isotonic media (100% SAR). The optimal conditions for the culture of Lim. tasmaniensis eggs were identified as a holding media of 100% SAR, followed by a medium change to 2.5% SAR at insemination. This sequence of media minimised the rate of swelling of the jelly layers prior to contact with the spermatozoa, and maximised the activation of spermatozoa and eggs throughout fertilisation and embryonic development. Embryos of Lim. tasmaniensis were cultured at four temperatures (13 C, 17 C, 23 C and 29 C), to determine the effect of temperature on cleavage and embryonic development rates. Embryonic development progressed through a sequence of stages that were not altered by changes in temperature. However cleavage rates were affected by changes in temperature as compared with normal embryonic growth at 23 C. Embryonic development was suspended at the lowest temperature (13 C) while embryonic viability was maintained. A moderate decrease in temperature (17 C) slowed cleavage, while the highest temperature (29 C) increased the cleavage rate, but decreased the embryo survival. Rates of embryonic development can be manipulated by changes in temperature and this method can be used to source blastomeres of a specific size/stage at a predetermined age or halt cleavage at specific stages for embryos or embryo derived cells to be included in ART procedures. This study produced the first report of the application of Intracytoplasmic Spermatozoon Injection (ICSI) in an Australian amphibian. Eggs that were activated by microinjection with a single spermatozoon (n=50) formed more deep, but abnormal, cleavage furrows post-injection (18/50, 36%), than surface changes (12/50, 24%). This result is in contrast to eggs injected without a spermatozoon (n=42), where the majority of eggs displayed limited surface changes (36/42, 86%), and few deep, abnormal furrows (3/42, 7%). Three advanced embryos (3/50, 6%) were produced by ICSI that developed to various stages within the culture system. Technical difficulties were encountered that prevented the generation of any metamorphs from ICSI tadpoles. Nevertheless, when these blocks to ICSI are overcome, the ICSI procedure will be both directly useful as an ART procedure in its own right, and the associated refinement of micromanipulation procedures will assist in the development of other ART procedures in Lim. tasmaniensis. A greater understanding of basic reproductive and developmental biology in Lim. tasmaniensis would greatly facilitate refinement of fertilisation by ICSI. Assisted Reproductive Technologies, in conjunction with gene banks may in the future regenerate extinct amphibian species, and assist in the recovery of declining amphibian populations nationally and worldwide.

Larger or broader : performance implications of size and diversity of the knowledge worker's egocentric network. [更多或更广：知识员工个人中心网络大小和 多样性对绩效的影响]

Management scholars and practitioners emphasize the importance of the size and diversity of a knowledge worker's social network. Constraints on knowledge workers’ time and energy suggest that more is not always better. Further, why and how larger networks contribute to valuable outcomes deserves further understanding. In this study, we offer hypotheses to shed insight on the question of the diminishing returns of large networks and the specific form of network diversity that may contribute to innovative performance among knowledge workers. We tested our hypotheses using data collected from 93 R&D engineers in a Sino-German automobile electronics company located in China. Study findings identified an inflection point, confirming our hypothesis that the size of the knowledge worker's egocentric network has an inverted U-shaped effect on job performance. We further demonstrate that network dispersion richness (the number of cohorts that the focal employee has connections to) rather than network dispersion evenness (equal distribution of ties across the cohorts) has more influence on the knowledge worker's job performance. Additionally, we found that the curvilinear effect of network size is fully mediated by network dispersion richness. Implications for future research on social networks in China and Western contexts are discussed.

A case-control study of ambient heat exposure and urolithiasis among outdoor workers in a shipbuilding company, Guangzhou, China

Higher ambient temperatures will increase heat stress on workers, leading to impacts upon their individual health and productivity. In particular, research has indicated that higher ambient temperatures can increase the prevalence of urolithiasis. This thesis examines the relationship between ambient heat exposure and urolithiasis among outdoor workers in a shipbuilding company in Guangzhou, China, and makes recommendations for minimising the possible impacts of high ambient temperatures on urolithiasis. A retrospective 1:4 matched case-control study was performed to investigate the association between ambient heat exposure and urolithiasis. Ambient heat exposure was characterised by total exposure time, type of work, department and length of service. The data were obtained from the affiliated hospital of the shipbuilding company under study for the period 2003 to 2010. A conditional logistic regression model was used to estimate the association between heat exposure and urolithiasis. This study found that the odds ratio (OR) of urolithiasis for total exposure time was 1.5 (95% confidence interval (CI): 1.2–1.8). Eight types of work in the shipbuilding company were investigated, including welder, assembler, production security and quality inspector, planing machine operator, spray painter, gas-cutting worker and indoor employee. Five out of eight types of work had significantly higher risks for urolithiasis, and four of the five mainly consisted of outdoors work with ORs of 4.4 (95% CI: 1.7–11.4) for spray painter, 3.8 (95% CI: 1.9–7.2) for welder, 2.7 (95% CI: 1.4–5.0) for production security and quality inspector, and 2.2 (95% CI: 1.1–4.3) for assembler, compared to the reference group (indoor employee). Workers with abnormal blood pressure (hypertension) were more likely to have urolithiasis with an OR of 1.6 (95% CI: 1.0–2.5) compared to those without hypertension. This study contributes to the understanding of the association between ambient heat exposure and urolithiasis among outdoor workers in China. In the context of global climate change, this is particularly important because rising temperatures are expected to increase the prevalence of urolithiasis among outdoor workers, putting greater pressure on productivity, occupational health management and health care systems. The results of this study have clear implications for public health policy and planning, as they indicate that more attention is required to protect outdoor workers from heat-related urolithiasis.

An introduction to biologics

An introduction to biologics 23.1 Introduction: Principles of biologics and their use as medicines 23.2 Protein biologics used as drugs 23.2.1 Proteins that function through enzymatic or regulatory activity. 23.2.1.1 Biologics as replacement of a deficient or abnormal protein. 23.2.1.2 Proteins that augment an existing biological process. 23.2.1.3 Proteins that provide a novel function or activity. 23.2.2. Proteins that function through specific targeting activity. 23.2.2.1. Monoclonal antibody nomenclature. 23.2.2.2. Naked monoclonal antibodies. 23.2.2.3. Conjugated monoclonal antibodies. 23.2.3. Recombinant protein vaccines.