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Apoptosis, a physiological process for the controlled deletion of cells, is critical for the regulation of cell numbers, the management of morphogenesis during embryonic development, and the orchestration of many cellular processes in the adult. Spermatogenesis, the production of functional spermatozoa from spermatogonial stem cells, is no exception. It appears that a functional apoptotic pathway is necessary for normal spermatogenesis to develop, and without it infertility ensues. Apoptosis also plays a crucial role in the maintenance of the testis and its response to external toxicants, as well as in the programmed senescence of terminally differentiated spermatozoa. This chapter focuses specifically on how apoptosis affects sperm quality and function, and the implications of this process for both embryonic development and the health and well-being of the offspring.
Our knowledge of the universe comes from recording the photon and particle fluxes incident on the Earth from space. We thus require sensitive measurement across the entire energy spectrum, using large telescopes with efficient instrumentation located on superb sites. Technological advances and engineering constraints are nearing the point where we are recording as many photons arriving at a site as is possible. Major advances in the future will come from improving the quality of the site. The ultimate site is, of course, beyond the Earth’s atmosphere, such as on the Moon, but economic limitations prevent our exploiting this avenue to the degree that the scientific community desires. Here we describe an alternative, which offers many of the advantages of space for a fraction of the cost: the Antarctic Plateau.
Intrauterine growth restriction (IUGR) and postnatal catch-up growth confer an increased risk of adult-onset disease. Overnourishment of adolescent ewes generates IUGR in ∼50% of lambs, which subsequently exhibit increased fractional growth rates. We investigated putative epigenetic changes underlying this early postnatal phenotype by quantifying gene-specific methylation at cytosine:guanine (CpG) dinucleotides. Hepatic DNA/RNA was extracted from IUGR [eight male (M)/nine female (F)] and normal birth weight (12 M/9 F) lambs. Polymerase chain reaction was performed using primers targeting CpG islands in 10 genes: insulin, growth hormone, insulin-like growth factor (IGF)1, IGF2, H19, insulin receptor, growth hormone receptor, IGF receptors 1 and 2, and the glucocorticoid receptor. Using pyrosequencing, methylation status was determined by quantifying cytosine:thymine ratios at 57 CpG sites. Messenger RNA (mRNA) expression of IGF system genes and plasma IGF1/insulin were determined. DNA methylation was independent of IUGR status but sexual dimorphism in IGF1 methylation was evident (M<F, P=0.008). IGF1 mRNA:18S and plasma IGF1 were M>F (both P<0.001). IGF1 mRNA expression correlated negatively with IGF1 methylation (r=−0.507, P=0.002) and positively with plasma IGF1 (r=0.884, P<0.001). Carcass and empty body weights were greater in males (P=0.002–0.014) and this gender difference in early body conformation was mirrored by sexual dimorphism in hepatic IGF1 DNA methylation, mRNA expression and plasma IGF1 concentrations.
The development of safe, effective, efficient methods for the isolation of human spermatozoa is a major feature of current research into the optimization of assisted conception therapy. DNA damage in spermatozoa has been linked with a wide variety of adverse clinical outcomes and a wide variety of defects in the offspring ranging from neurological conditions such as autism or spontaneous schizophrenia to childhood cancer. Although definitive evidence is lacking, it is possible that DNA damage in the male germ line makes a significant contribution to the elevated incidence of birth defects observed following assisted reproductive technology (ART), when compared with the naturally conceived population. The spermatozoa can be isolated from semen using discontinuous gradient centrifugation or a new electrophoretic approach. Preliminary results suggest that the latter represents an extremely rapid, effective means of isolating high quality cells for insemination. Additional clinical trials are now needed to substantiate this early promise.
This chapter focuses specifically on how apoptosis affects sperm quality and function, and the implications of this process for both embryonic development and the health and well-being of the offspring. DNA damage in human spermatozoa has been correlated with poor fertilization and impaired embryonic development to the blastocyst stage as well as with the incidence of subsequent miscarriage. Human infertility is a complex multifactorial condition that is strongly impacted by genetic factors that assisted reproductive technology (ART) will ensure are passed onto the progeny. Spermiogenesis is a key event in the etiology of DNA damage in the male germ line. DNA damage in human spermatozoa appears to have its origins in the testes and is associated with oxidative stress. Spermatozoa possess several variants of the prolactin receptor and respond to the presence of this hormone with the stimulation of PI3 kinase/Akt phosphorylation and the prolongation of sperm survival.
Recent events have heightened awareness of disaster health issues and the need to prepare the health workforce to plan for and respond to major incidents. This has been reinforced at an international level by the World Association for Disaster and Emergency Medicine, which has proposed an international educational framework.
The aim of this paper is to outline the development of a national educational framework for disaster health in Australia.
The framework was developed on the basis of the literature and the previous experience of members of a National Collaborative for Disaster Health Education and Research. The Collaborative was brought together in a series of workshops and teleconferences, utilizing a modified Delphi technique to finalize the content at each level of the framework and to assign a value to the inclusion of that content at the various levels.
The framework identifies seven educational levels along with educational outcomes for each level. The framework also identifies the recommended contents at each level and assigns a rating of depth for each component. The framework is not intended as a detailed curriculum, but rather as a guide for educationalists to develop specific programs at each level.
This educational framework will provide an infrastructure around which future educational programs in Disaster Health in Australia may be designed and delivered. It will permit improved articulation for students between the various levels and greater consistency between programs so that operational responders may have a consistent language and operational approach to the management of major events.
In Expt 1, 34 individually-penned Finn Dorset ewes of mean live weight 68 kg were synchronized in oestrus and mated to Suffolk rams. From mating until day 28 of pregnancy each received daily 15 MJ of metabolizable energy (ME) and 225 g crude protein (CP). From day 28 to slaughter on days 34, 41, 48 or 55 half of the ewes continued on this feeding regime and half had their daily intake reduced abruptly to 7·5 MJ of ME and 112 g CP. The mean number of ovulations per ewe was 4·03 (range 2–8) and the mean number of viable foetuses at time of slaughter 3·35 (range 2–6). The combined loss of ova (fertilization failure and early embryonic death) was 14·6% and detectable foetal deaths 2·2%. Level of feeding had no significant effect on these measures or on foetal growth. Foetal growth from 34 to 55 days was described by the equation
In w = 0·962–18·613 e-0·0272t–0·00091t(f–3),
where w = foetal weight (kg), t = age (days) and f = litter size. Within-litter variability measured as the S.D. of In w (kg) was 0–081 for twins, 0·108 for triplets and 0·106 for quadruplets and higher multiples.
In a second experiment Suffolk × Finn Dorset embryos were transplanted at the rate of two per uterine horn into 15 recipient Finn Dorset ewes. Embryo survival was 72% and foetal weights at 60 days varied from 67 to 146% of the mean value of 66 g. Withinlitter variation in foetal size was only about 70% of that expected for foetuses developing from the variable distribution in their initial positioning that occurs naturally. The correlation between foetal weight and placental weight at day 60 was 0·72 (P < 0001) indicating that the association between foetal weight and placental weight in prolific ewes is not confined to late pregnancy.
The results of both experiments are consistent with the hypothesis that the greater within-litter variability in birth weight in large litters is controlled by events in early pregnancy.
Background. Cognitive performance was compared in the genetically and neurobiologically related disorders of Tourette's syndrome (TS) and obsessive–compulsive disorder (OCD), in three domains of executive function: planning, decision-making and inhibitory response control.
Method. Twenty TS patients, twenty OCD patients and a group of age- and IQ-matched normal controls completed psychometric and computerized cognitive tests and psychiatric rating scales. The cognitive tests were well-characterized in terms of their sensitivity to other fronto-striatal disorders, and included pattern and spatial recognition memory, attentional set-shifting, and a Go/No-go set-shifting task, planning, and decision-making.
Results. Compared to controls, OCD patients showed selective deficits in pattern recognition memory and slower responding in both pattern and spatial recognition, impaired extra-dimensional shifting on the set-shifting test and impaired reversal of response set on the Go/No-go test. In contrast, TS patients were impaired in spatial recognition memory, extra-dimensional set-shifting, and decision-making. Neither group was impaired in planning. Direct comparisons between the TS and OCD groups revealed significantly different greater deficits for recognition memory latency and Go/No-go reversal for the OCD group, and quality of decision-making for the TS group.
Conclusions. TS and OCD show both differences (recognition memory, decision-making) and similarities (set-shifting) in selective profiles of cognitive function. Specific set-shifting deficits in the OCD group contrasted with their intact performance on other tests of executive function, such as planning and decision-making, and suggested only limited involvement of frontal lobe dysfunction, possibly consistent with OCD symptomatology.
In the tammar wallaby (Macropus eugenii), post-testicular acrosomal shaping involves a complex infolding
and fusion of the anterior and lateral projections of the scoop-shaped acrosome into a compact button-like
structure occupying the depression on the anterior end of the sperm nucleus. The present study has
generated cytochemical and histological evidence to demonstrate that the occurrence of actin filaments
(F-actin, labelled by Phalloidin-FITC) in the acrosome of tammar wallaby spermatozoa is temporally and
spatially associated with the process of acrosomal shaping in the epididymis, through a pool of monomeric
actin (G-actin, labelled by Rh-DNase I) present in the acrosome throughout all stages of epididymal
maturation. F-actin was not detected in the acrosome of testicular spermatozoa, but was found in the
infolding and condensing acrosome of caput and corpus epididymal spermatozoa. When the spermatozoa
completed acrosome shaping in the cauda epididymidis, F-actin disappeared from the acrosomal area. The
strong correlation between the occurrence of F-actin and the events of acrosomal shaping suggested that the
post-testicular shaping of the acrosome might depend on a precise succession of assembly and disassembly
of F-actin within the acrosome as the spermatozoa transit the epididymis. Thus, actin filaments might play a
significant role in the acrosomal transformation, as they are commonly involved in morphological changes in
The objective was to determine the effect of previous lambing date and subsequent month of mating on reproductive performance in Mule (Bluefaced Leicester × Scottish Blackface) ewes. Sixty-four ewes which had previously lambed in January (13 January (s.e. = 1 day)) and 80 ewes which had previously lambed in May (15 May (s.e. = 1 day)) were allocated equally to four mating periods (30 August to 17 September, 1 November to 19 November, 3 January to 21 January and 14 February to 4 March) in a 2 × 4 factorial design. From 20 days before and during their designated mating period, January- and May-lambing ewes were separately housed in straw-bedded pens under natural photoperiod and were given 1 kg per head per day dried grass pellets. A vasectomized ram was continuously present with each group for 17 days and was replaced by a raddled, fertility tested entire Suffolk ram at the start of the mating period. Ewes were mated at a single natural oestrus and those marked by the ram were recorded daily. Ovulation rate was measured by laparoscopy on day 6 after mating. For ewes which had previously lambed in January (16 per group), numbers by month of mating that showed oestrous behaviour, ovulated and were pregnant, respectively, were: September, 16, 15 and 12; November, 16, 15 and 14; January, 15, 15 and 10 and February, 15, 16 and 7. Mean (s.e.) ovulation rates by month of mating were 2·1 (0.16), 2·5 (0.19), 2·1 (0.09) and 2·2 (0.19) corpora lutea per ewe ovulating, and lambing rates by month of mating were 1·3 (0.25), 1·9 (0.25), 1·2 (0.24) and 0·8 (0.23) lambs per ewe to the ram. For ewes which had previously lambed in May (20 per group), numbers by month of mating that showed oestrous behaviour, ovulated and were pregnant, respectively, were: September, 13, 20 and 12; November, 20, 20 and 19; January, 20, 20 and 17 and February, 20, 20 and 13. Mean (s.e.) ovulation rates by month of mating were 2·0 (0.13), 2·3 (0.11), 2·1 (0.05) and 2·2 (0.11) corpora lutea per ewe ovulating, and lambing rates by month of mating were 1·0 (0.21), 2·1 (0.15), 1·5 (0.17) and 1·2 (0.21) lambs per ewe to the ram. Ovulation, pregnancy and lambing rates were not influenced by previous lambing date, but lambing rates were significantly (P < 0·01) reduced for ewes mated in September and February compared with November. Results demonstrate that in Mule ewes acceptable ovulation rates can be achieved throughout the period September to February but lambing rates are reduced when ewes are mated at the extremes of their natural breeding season. The main factor contributing to the reduction in lambing rates was an increase in the number of ewes failing to establish pregnancy as a consequence of ovulation without oestrous behaviour, fertilization failure and (or) total embryo loss.
The effect of lambing date on the subsequent onset and duration of ovarian cyclicity in Mule (Bluefaced Leicester × Scottish Blackface) ewes was investigated. Nineteen ewes which had lambed in January (16 January 1993 (s.e. 3 days)) and been weaned in February-March and 22 comparable ewes which had lambed in May (14 May 1993 (s.e. 2 days)) and been weaned on 23 August were maintained at pasture as two isolated groups. A raddled vasectomized ram was continually present with each group from 14 July 1993 to 26 May 1994 and marked (oestrous) ewes were recorded twice weekly. Ovarian activity was assessed by measuring peripheral progesterone concentrations in blood samples collected twice weekly and by laparoscopic viewing of the ovaries of all ewes during October, January and March. The onset and duration of ovarian activity were significantly affected by the previous lambing date. For January and May lambing ewes, mean dates of onset were 5 September 1993 (s.e. 2 days) v. 25 September 1993 (s.e. 4 days) (P < 0·001) and of cessation were 5 April 1994 (s.e. 5 days) v. 10 April 1994 (s.e. 3 days). Mean durations of ovarian activity were 212 (s.e. 6) and 195 (s.e. 5) days (P < 0·05) during which 12·4 (s.e. 0·29) and 11·5 (s.e. 0·38) ovarian cycles respectively were recorded. Ovulation rate was not affected by previous lambing date but was significantly lower in March compared with October (January lambing ewes 1·7 (s.e. 0·1) v. 2·3 (s.e. 0·1) (P < 0·001); May lambing ewes 1·6 (s.e. 0·1) v. 2·1 (s.e. 0·1) (P < 0·01)). Results demonstrate that (i) Mule ewes have a potential breeding season of up to 8 months duration; (ii) the onset and duration of ovarian activity can be influenced by previous lambing date; and (Hi) a seasonal decline in ovulation rate may, in practical terms, result in a lower lambing percentage for animals bred towards the end of their natural breeding period.
Priming with a progestagen-impregnated vaginal pessary followed by an injection of 750 i.u. pregnant mare serum gonadotropin at pessary withdrawal was used to induce oestrus and ovulation during anoestrus (13 June, latitude 57°N) in 58 Border Leicester × Scottish Blackface ewes that had lambed previously in late March/early April. At 54 to 58 h after pessary withdrawal the uterus of each ewe was viewed by laparoscopy and 50 × 106 viable sperm deposited into each uterine horn. Mean ovulation rate was 2·9 (s.d. 0·12; range 1 to 5). Plasma progesterone concentrations remained >1·5 μg/l in 54 (93%) of the ewes until at least day 21 after insemination implying a high fertilization rate. After allowing for three ewes that were observed to abort between 39 and 55 days after insemination and another three, carrying nine normally developed lambs, that died between 102 and 136 days of gestation, 40 of the remaining 48 ewes (i.e. 69% of the original 58) produced 76 viable lambs in early November. These conception and lambing rates obtained following the intrauterine insemination of 100× 106 viable sperm per ewe are more than double those obtained following natural mating or conventional cervical insemination with four times the number of viable sperm and imply that intrauterine insemination may enhance lamb production from ewes induced to breed during seasonal anoestrus.