Nucleus downscaling in mouse embryos is regulated by cooperative developmental and geometric programs

Maintaining appropriate nucleus size is important for cell health, but the mechanisms by which this is achieved are poorly understood. Controlling nucleus size is a particular challenge in early development, where the nucleus must downscale in size with progressive reductive cell divisions. Here we use live and fixed imaging, micromanipulation approaches, and small molecule analyses during preimplantation mouse development to probe the mechanisms by which nucleus size is determined. We find a close correlation between cell and nuclear size at any given developmental stage, and show that experimental cytoplasmic reduction can alter nuclear size, together indicating that cell size helps dictate nuclear proportions. Additionally, however, by creating embryos with over-sized blastomeres we present evidence of a developmental program that drives nuclear downscaling independently of cell size. We show that this developmental program does not correspond with nuclear import rates, but provide evidence that PKC activity may contribute to this mechanism. We propose a model in which nuclear size regulation during early development is a multi-mode process wherein nucleus size is set by cytoplasmic factors, and fine-tuned on a cell-by-cell basis according to cell size

Anaphase B precedes anaphase A in the mouse egg.

Segregation of chromosomes at the time of cell division is achieved by the microtubules and associated molecules of the spindle. Chromosomes attach to kinetochore microtubules (kMTs), which extend from the spindle pole region to kinetochores assembled upon centromeric DNA. In most animal cells studied, chromosome segregation occurs as a result of kMT shortening, which causes chromosomes to move toward the spindle poles (anaphase A). Anaphase A is typically followed by a spindle elongation that further separates the chromosomes (anaphase B). The experiments presented here provide the first detailed analysis of anaphase in a live vertebrate oocyte and show that chromosome segregation is initially driven by a significant spindle elongation (anaphase B), which is followed by a shortening of kMTs to fully segregate the chromosomes (anaphase A). Loss of tension across kMTs at anaphase onset produces a force imbalance, allowing the bipolar motor kinesin-5 to drive early anaphase B spindle elongation and chromosome segregation. Early anaphase B spindle elongation determines the extent of chromosome segregation and the size of the resulting cells. The vertebrate egg therefore employs a novel mode of anaphase wherein spindle elongation caused by loss of k-fiber tension is harnessed to kick-start chromosome segregation prior to anaphase A

Evaluation of alternative intersection treatments at rural crossroads using simulation software

Objective: Rural roads are characterised by hazardous roadsides and suboptimal geometry, yet allow for high travel speeds and unfavourable impact angles. In Victoria, 25% of persons seriously injured and 52% of fatalities occur on rural roads, with 30% occurring at intersections (Transport Accident Commision, 2016; Victorian Parliament Road Safety Committee 2002). In the USA, almost twice the number of traffic fatalities occur in rural areas than in urban areas, whilst accounting for less than half of all vehicle miles travelled and 21% of the population (NHTSA 2002). The choice of safety countermeasure is therefore paramount. Simulation software provides a cost-effective means of analysing alternative intersection treatments with a view to identifying their effectiveness in mitigating crashes. The aim of this research was to assess the safety benefits of four alternative intersection treatments using in-depth crash data with an advanced crash reconstruction process. Method: Using a single serious injury real-world crash from the MUARC Enhanced Crash Investigation Study (ECIS, Fitzharris et al., 2015) and crash reconstruction software, an exemplar rural crash was reconstructed and validated against real-world data. The crash involved a passenger vehicle (EuroNCAP 5*) approaching from a minor road and failing to yield at a ‘give-way’ sign; the posted speed limit was 80 km/h. The vehicle was struck on the right/driver side by a rigid truck (B-vehicle; 1990) travelling on the major approach (100 km/h). The driver of the case vehicle was seriously injured. Four alternative intersection treatments appropriate for the crash site were constructed in CAD software (Rhinoceros V5): roundabout; rumble strips; a reduced speed limit and the combination of lower speed limit and rumbles to determine the reduction in crash forces in the presence of the countermeasures. Results: The hypothetical scenarios demonstrate substantial reductions in impact force and different points of impact, resulting in a significantly lower injury severity for the struck driver. Speed limit reduction to 80 km/h on the main approach (from 100 km/h) in combination with rumble strips on both intersection approaches had the most favorable outcome with the crash avoided entirely, assuming speed compliance. Discussion: The findings have implications for understanding the role of speed in crashes, and hence the design of effective countermeasures. Simulation software, validated using real-world data, provides a cost effective means of evaluating alternative intersection treatments for rural intersections. Scaled up, implementing these treatments would have significant safety benefits and reduce the road trauma currently associated with rural roads

The MUARC-TAC enhanced crash investigation study: study update, analysis of crash types and contributing factors [Abstract]

This paper presents an update of the Monash University Accident Research Centre (MUARC) – Transport Accident Commission (TAC) Enhanced Crash Investigation Study (ECIS) as well as an exploration of the characteristics of injured drivers, crash types and factors implicated in crash occurrence. Three configurations are of particularly high frequency and severity, whilst crashes involving young and older drivers are different in nature and have different contributing factors. Fatigue, driver error, and pre-crash driver blackouts due to medical conditions were prominent contributing factors. Injury severity would be significantly lower in 32% of cases if either front or side airbags were fitted. The findings point to key risk factors that can be addressed in road safety strategies

What drivers do while speeding: examining the associations between speeding and driver distraction through the Enhanced Crash Investigation Study protocol [Abstract]

This paper represents an exploratory analysis to assess the feasibility of assessing the relationship between driver speed and engagement in potentially distracting behaviours. Control data from the ECIS project are examined. These data include both objective speed measurements recorded via laser camera positioned at ECIS case-vehicle crash locations as well as retrospective self-reported driving behaviours from drivers recorded at these sites. Exploratory analysis suggests that the activities reported by drivers with recorded speeds above the limit may differ from the activities reported by those recorded on or below the limit

The MUARC-TAC enhanced crash investigation study: a platform to understand the causes and consequences of serious injury crashes.

Background: In recognising the consequences of serious injury crashes, the Transport Accident Commission (TAC) commissioned Monash University Accident Research Centre (MUARC) to undertake the Enhanced Crash Investigation Study (ECIS). This paper describes the program components, seven key research questions and technical innovations used in the study. We describe the information collected and outline a ‘Safe Systems Failure Analysis’ used for each case. Project Method: Participants in ECIS include drivers aged 18 years and older seriously injured in crashes on public Victorian roads. Drivers are recruited whilst inpatients at a major trauma hospital and where possible interviews conducted. The ECIS team inspects their crashed vehicle and critically analyses the crash environment. Event Data Recorder (EDR, black-box) data is acquired from vehicles where possible and crash reconstructions are undertaken. Each case is submitted to an internal panel review with a sub-sample of cases presented to external panels throughout Victoria. This process leads to each case being submitted to a Safe Systems Failure Analysis where contributing factors and countermeasures are identified by a broad group of stakeholders. The ECIS control arm permits examination of the relationship between certain factors, such as speed and crash occurrence. Results and Discussion: In addition to describing the study, we provide an example of how the identification of crash factors, using a Safe Systems paradigm based on real-world serious injury crashes, can lead to the identification of targeted countermeasures, each with an identified policy action. Implications: This paper will demonstrate a method for creating a robust evidence base upon which government road safety policy can be built. By scaling up individual crash findings to the broader crash population, countermeasures and associated policy actions can be appropriately prioritised

An assessment of the Jenkinson and Collison synoptic classification to a continental mid-latitude location

A weather-type catalogue based on the Jenkinson and Collison method was developed for an area in south-west Russia for the period 1961--2010. Gridded sea level pressure data was obtained from the National Centers for Environmental Prediction/National Center for Atmospheric Research (NCEP/NCAR) reanalysis. The resulting catalogue was analysed for frequency of individual types and groups of weather types to characterise long-term atmospheric circulation in this region. Overall, the most frequent type is anticyclonic (A) (23.3 {%}) followed by cyclonic (C) (11.9 {%}); however, there are some key seasonal patterns with westerly circulation being significantly more common in winter than summer. The utility of this synoptic classification is evaluated by modelling daily rainfall amounts. A low level of error is found using a simple model based on the prevailing weather type. Finally, characteristics of the circulation classification are compared to those for the original JC British Isles catalogue and a much more equal distribution of flow types is seen in the former classification

Crash characteristics and patterns of injury among hospitalized motorised two-wheeled vehicle users in urban India

<p>Abstract</p> <p>Background</p> <p>Traffic crashes and consequent injuries represent a growing public health concern in India, particularly in light of increasing motorization. Motorised two-wheeled vehicles (MTV) constitute a large portion of the vehicle fleet in India. We report the crash characteristics and injury patterns among a cohort of MTV riders and pillions presenting to hospital post-crash.</p> <p>Methods</p> <p>Consecutive MTV riders and pillions, whether alive or dead, injured in a road traffic crash presenting to the emergency departments of two government hospitals and three branches of a private hospital in urban Hyderabad, India, were recruited to this study.</p> <p>Results</p> <p>378 MTV users were enrolled to the study of whom 333 (88.1%) were male, 252 (66.7%) were riders and median age was 31.3 years. A total of 223 (59%) MTV users were injured in multi-vehicle crashes while one-third had a frontal impact. The majority (77%) were assessed as having a Glasgow coma score (GCS) of 13–15, 12% a GCS of 9–12 and 11% a GCS of 3–8. No difference was seen in the severity distribution of injuries based on GCS among riders and pillions. Open wounds and superficial injuries to the head (69.3%) and upper extremity (27%) and lower extremity (24%) were the most common injuries. 43 (11%) sustained an intracranial injury, including 12 (28%) with associated fracture of the bones of the head. There were few differences in types of injuries sustained by riders and pillions though riders had a significantly lower risk of crush injuries of the lower extremity than pillions (relative risk, RR 0.25, 95% CI 0.08–0.81) and female pillions were at a significantly lower risk of sustaining fractures of the lower extremity than male pillions (RR 0.30, 95% CI 0.09 – 0.94). Overall, 42 (11%) MTV users died, of which 42.8% died before reaching the hospital. Only 74 (19.6%) MTV users had worn a helmet correctly and failure to wear a helmet was associated with a five times greater risk of intracranial injury (RR 4.99, 95% CI 1.23–20.1). Of the 19 pre-hospital deaths, 16 (84%) had not worn a helmet.</p> <p>Conclusion</p> <p>Head injuries accounted for the major proportion of injuries sustained in MTV users. Non-helmet use was associated with increased risk of serious head injuries. The data presented on the nature and severity of injuries sustained by MTV users can assist with planning to deal with these consequences as well as prevention of these injuries given the high use of MTV in India.</p

Oocyte–Targeted Deletion Reveals That Hsp90b1 Is Needed for the Completion of First Mitosis in Mouse Zygotes

Hsp90b1 is an endoplasmic reticulum (ER) chaperone (also named Grp94, ERp99, gp96,Targ2, Tra-1, Tra1, Hspc4) (MGI:98817) contributing with Hspa5 (also named Grp78, BIP) (MGI:95835) to protein folding in ER compartment. Besides its high protein expression in mouse oocytes, little is known about Hsp90b1 during the transition from oocyte-to-embryo. Because the constitutive knockout of Hsp90b1 is responsible for peri-implantation embryonic lethality, it was not yet known whether Hsp90b1 is a functionally important maternal factor.To circumvent embryonic lethality, we established an oocyte-specific conditional knockout line taking advantage of the more recently created floxed Hsp90b1 line (Hsp90b1(flox), MGI:3700023) in combination with the transgenic mouse line expressing the cre recombinase under the control of zona pellucida 3 (ZP3) promoter (Zp3-cre, MGI:2176187). Altered expression of Hsp90b1 in growing oocytes provoked a limited, albeit significant reduction of the zona pellucida thickness but no obvious anomalies in follicular growth, meiotic maturation or fertilization. Interestingly, mutant zygotes obtained from oocytes lacking Hsp90b1 were unable to reach the 2-cell stage. They exhibited either a G2/M block or, more frequently an abnormal mitotic spindle leading to developmental arrest. Despite the fact that Hspa5 displayed a similar profile of expression as Hsp90b1, we found that HSPA5 and HSP90B1 did not fully colocalize in zygotes suggesting distinct function for the two chaperones. Consequently, even if HSPA5 was overexpressed in Hsp90b1 mutant embryos, it did not compensate for HSP90B1 deficiency. Finally, further characterization of ER compartment and cytoskeleton revealed a defective organization of the cytoplasmic region surrounding the mutant zygotic spindle.Our findings demonstrate that the maternal contribution of Hsp90b1 is critical for the development of murine zygotes. All together our data indicate that Hsp90b1 is involved in unique and specific aspects of the first mitosis, which brings together the maternal and paternal genomes on a single spindle

Injury in China: a systematic review of injury surveillance studies conducted in Chinese hospital emergency departments

<p>Abstract</p> <p>Background</p> <p>Injuries represent a significant and growing public health concern in China. This <it>Review </it>was conducted to document the characteristics of injured patients presenting to the emergency department of Chinese hospitals and to assess of the nature of information collected and reported in published surveillance studies.</p> <p>Methods</p> <p>A systematic search of MEDLINE and China Academic Journals supplemented with a hand search of journals was performed. Studies published in the period 1997 to 2007 were included and research published in Chinese was the focus. Search terms included emergency, injury, medical care.</p> <p>Results</p> <p>Of the 268 studies identified, 13 were injury surveillance studies set in the emergency department. Nine were collaborative studies of which eight were prospective studies. Of the five single centre studies only one was of a prospective design. Transport, falls and industrial injuries were common mechanisms of injury. Study strengths were large patient sample sizes and for the collaborative studies a large number of participating hospitals. There was however limited use of internationally recognised injury classification and severity coding indices.</p> <p>Conclusion</p> <p>Despite the limited number of studies identified, the scope of each highlights the willingness and the capacity to conduct surveillance studies in the emergency department. This <it>Review </it>highlights the need for the adoption of standardized injury coding indices in the collection and reporting of patient health data. While high level injury surveillance systems focus on population-based priority setting, this <it>Review </it>demonstrates the need to establish an internationally comparable trauma registry that would permit monitoring of the trauma system and would by extension facilitate the optimal care of the injured patient through the development of informed quality assurance programs and the implementation of evidence-based health policy.</p