Traversing the ovine cervix - a challenge for cryopreserved semen and creative science

This review brings together research findings on cervical relaxation in the ewe and its pharmacological stimulation for enhancement of the penetration needed for transcervical insemination and embryo transfer. On the basis that the success of artificial insemination is the percentage of ewes lambing, a review is made of recent research aimed at understanding and minimising the sub-lethal effects of freezing and thawing on the viability of spermatozoa, their membrane integrity and their ability to migrate through cervical mucus, as these characteristics have a major influence on fertility, particularly when semen is deposited, artificially, in the os cervix. Milestones of achievement are given for transcervical intrauterine insemination, embryo recovery and transfer and the birth of lambs of pre-determined sex, firstly following intracytoplasmic sperm injection, then laparoscopic intrauterine insemination using highly diluted flow-cytometrically sorted fresh semen and subsequently by os cervix insemination using sexed semen that had been frozen and thawed. Diversity of research endeavour (applied, cellular, molecular), research discipline (anatomy, histology, immunology, endocrinology) and research focus (cell, tissue, organ, whole animal) is embraced within the review as each has significant contributions to make in advancing recent scientific findings from the laboratory into robust on-farm transcervical insemination and embryo transfer techniques.     

Natal movement in juvenile Atlantic salmon: a body size-dependent strategy?

If competitive ability depends on body size, then the optimal natal movement from areas of high local population density can also be predicted to be size-dependent. Specifically, small, competitively-inferior individuals would be expected to benefit most from moving to areas of lower local density. Here we evaluate whether individual variation in natal movement following emergence from nests is consistent with such a size-dependent strategy in Atlantic salmon, and whether such a strategy is evident across a range of environmental conditions (principally predator presence and conspecific density). In stream channel experiments, those juveniles that stayed close to nests were larger than those that emigrated. This result was not sensitive to predator presence or conspecific density. These observations were mirrored in natural streams in which salmon eggs were planted in nests and the resulting offspring were sampled at high spatial resolution. A negative relationship was found between juvenile body size and distance from nests early in development whereas in those streams sampled later in ontogeny, individuals that had moved furthest were largest. Thus, movement away from nests appeared to result in a reduced competitive intensity and increased growth rate. The fact that there is ultimately a growth advantage associated with moving suggests that there is also a cost that selects against movement by the larger individuals. Thus, natal movement in juvenile Atlantic salmon appears to represent a body size-dependent strategy.     

In vitro glucocorticoid sensitivity is associated with clinical glucocorticoid therapy outcome in rheumatoid arthritis

Introduction

Genetic and disease-related factors give rise to a wide spectrum of glucocorticoid (GC) sensitivity in rheumatoid arthritis (RA). In clinical practice, GC treatment is not adapted to these differences in GC sensitivity. In vitro assessment of GC sensitivity before the start of therapy could allow more individualized GC therapy. The aim of the study was to investigate the association between in vitro and in vivo GC sensitivity in RA.

Methods

Thirty-eight early and 37 established RA patients were prospectively studied. In vitro GC sensitivity was assessed with dexamethasone-induced effects on interleukin-2 (IL-2) and glucocorticoid-induced leucine zipper (GILZ) messenger RNA expression in peripheral blood mononuclear cells (PBMCs). A whole-cell dexamethasone-binding assay was used to measure number and affinity (1/K_D) of glucocorticoid receptors (GRs).In vivo GC sensitivity was determined by measuring the disease activity score (DAS) and health assessment questionnaire disability index (HAQ-DI) score before and after 2 weeks of standardized GC treatment.

Results

GR number was positively correlated with improvement in DAS. IL-2-EC₅₀ and GILZ-EC₅₀ values both had weak near-significant correlations with clinical improvement in DAS in intramuscularly treated patients only. HAQ responders had lower GILZ-EC₅₀ values and higher GR number and K_D.

Conclusions

Baseline cellular in vitro glucocorticoid sensitivity is modestly associated with in vivo improvement in DAS and HAQ-DI score after GC bridging therapy in RA. Further studies are needed to evaluate whether in vitro GC sensitivity may support the development of tailor-made GC therapy in RA.     

Sensitivity to cell killing and the induction of cytogenetic damage following gamma irradiation in wild-type and thymidine kinase-deficient Friend mouse erythroleukaemia cells

Wild-type Friend erythroleukaemia (clone 707) cells and 2 thymidine kinase-deficient subclones, 707BUE and 707BUF, having thymidine kinase activities of 1.4% and 0.7% that of clone 707, were compared for sensitivity to killing and the induction of cytogenetic damage following irradiation. Three doses of gamma irradiation were used (150, 300 and 450 cGy), and cells were harvested for metaphase spreads after 4, 8, 12, 15, 29 and 43 h. G₂ delay was evident at 4 h following gamma irradiation in the 3 cell clones examined, and recovery of mitosis was observed to be dose-dependent. G₂ delay was found to be most prolonged in subclone 707BUE and most prompt in clone 707. Increased sensitivity to the induction of cytogenetic aberrations at all three doses was apparent in the 2 thymidine kinase-deficient subclones (as compared to wild-type cells) at 15, 29 and 43 h. Th thymidine kinase-deficient subclones also showed increased sensitivity to gamma radiation-induced cell killing. Furthermore, subclone 707BUE consistently exhibited greater to gamma irradiation than did the subclone with lower thymidine kinase activity, 707BUF. The importance of thymidine kinase levels and extended G₂ delay for DNA repair processes is discussed.     

Relative shortening and functional tethering of spinal cord in adolescent scoliosis – Result of asynchronous neuro-osseous growth, summary of an electronic focus group debate of the IBSE

There is no generally accepted scientific theory for the causes of adolescent idiopathic scoliosis (AIS). As part of its mission to widen understanding of scoliosis etiology, the International Federated Body on Scoliosis Etiology (IBSE) introduced the electronic focus group (EFG) as a means of increasing debate on knowledge of important topics. This has been designated as an on-line Delphi discussion. The Statement for this debate was written by Dr WCW Chu and colleagues who examine the spinal cord to vertebral growth interaction during adolescence in scoliosis. Using the multi-planar reconstruction technique of magnetic resonance imaging they investigated the relative length of spinal cord to vertebral column including ratios in 28 girls with AIS (mainly thoracic or double major curves) and 14 age-matched normal girls. Also evaluated were cerebellar tonsillar position, somatosensory evoked potentials (SSEPs), and clinical neurological examination. In severe AIS compared with normal controls, the vertebral column is significantly longer without detectable spinal cord lengthening. They speculate that anterior spinal column overgrowth relative to a normal length spinal cord exerts a stretching tethering force between the two ends, cranially and caudally leading to the initiation and progression of thoracic AIS. They support and develop the Roth-Porter concept of uncoupled neuro-osseous growth in the pathogenesis of AIS which now they prefer to term ' asynchronous neuro-osseous growth'. Morphological evidence about the curve apex suggests that the spinal cord is also affected, and a 'double pathology' is suggested. AIS is viewed as a disorder with a wide spectrum and a common neuroanatomical abnormality namely, a spinal cord of normal length but short relative to an abnormally lengthened anterior vertebral column. Neuroanatomical changes and/or abnormal neural function may be expressed only in severe cases. This asynchronous neuro-osseous growth concept is regarded as one component of a larger concept. The other component relates to the brain and cranium of AIS subjects because abnormalities have been found in brain (infratentorial and supratentorial) and skull (vault and base). The possible relevance of systemic melatonin-signaling pathway dysfunction, platelet calmodulin levels and putative vertebral vascular biology to the asynchronous neuro-osseous growth concept is discussed. A biomechanical model to test the spinal component of the concept is in hand. There is no published research on the biomechanical properties of the spinal cord for scoliosis specimens. Such research on normal spinal cords includes movements (kinematics), stress-strain responses to uniaxial loading, and anterior forces created by the stretched cord in forward flexion that may alter sagittal spinal shape during adolescent growth. The asynchronous neuro-osseous growth concept for the spine evokes controversy. Dr Chu and colleagues respond to five other concepts of pathogenesis for AIS and suggest that relative anterior spinal overgrowth and biomechanical growth modulation may also contribute to AIS pathogenesis.     

Maintaining diversity in an artificially propagated population

Populations may suffer unexpected loss or distortion of biodiversity as a consequence of strategies employed in artificial propagation programs. The Trinity River Fish Hatchery may have inadvertently experienced this while attempting to preserve diversity in a return time within a Chinook salmon population. We develop a model for this system and prove that the long-term distribution of return types converges and that it is strongly tied to the management strategy. Given estimates of heritabilities for return type and differential survival rates, an estimate of this long-term distribution can be computed easily.     

Non-target effects of an introduced biological control agent on deer mouse ecology

Release of exotic insects as biological control agents is a common approach to controlling exotic plants. Though controversy has ensued regarding the deleterious direct effects of biological control agents to non-target species, few have examined the indirect effects of a ”well-behaved” biological control agent on native fauna. We studied a grassland in west-central Montana infested with spotted knapweed (Centaurea maculosa) to examine the effects of knapweed invasion and two gall flybiological control agents (Urophora affinis and U. quadrifasciata) on the native deer mouse (Peromyscus maniculatus). Stomach-content analysis revealed that Urophora were the primary food item in Peromyscus diets for most of the year and made up 84–86% of the winter diet. Stomach contents indicated that wild-caught mice consumed on average up to 247 Urophora larvae mouse^–1 day^–1, while feeding trials revealed that deer mice could depredate nearly 5 times as many larvae under laboratory conditions. In feeding trials, deer mice selected knapweed seedheads with greater numbers of galls while avoiding uninfested seedheads. When Urophora larvae were present in knapweed seedheads, deer mice selected microhabitats with moderately high (31–45% cover) and high knapweed infestation (≥46% cover). After Urophora emerged and larvae were unavailable to Peromyscus, mice reversed habitat selection to favor sites dominated by native-prairie with low knapweed infestation (0–15%). Establishment of the biological control agent, Urophora spp., has altered deer mouse diets and habitat selection by effecting changes in foraging strategies. Deer mice and other predators may reduce Urophora populations below a threshold necessary to effectively control spotted knapweed. Received: 04 May 1999 / Accepted: 14 August 1999     

Response of Dendroctonus brevicomis to different release rates of nonhost angiosperm volatiles and verbenone in trapping and tree protection studies

A blend of eight nonhost angiosperm volatiles (benzyl alcohol, benzaldehyde, guaiacol, nonanal, salicylaldehyde, (E)‐2‐hexenal, (E)‐2‐hexen‐1‐ol and (Z)‐2‐hexen‐1‐ol) without [NAV] and with [NAVV] (–)‐verbenone (4,6,6‐trimethylbicyclo[3.1.1]hept‐3‐en‐2‐one) were tested at low (L), medium (M) and high (H) release rates for their ability to reduce attraction of western pine beetle, Dendroctonus brevicomis LeConte, to attractant‐baited (exo‐brevicomin [racemic, 3 mg/d], frontalin [racemic, 3 mg/d] and myrcene [18 mg/d]) multiple funnel traps. NAV‐L (40 mg/d) had no significant effect. Verbenone alone (50 mg/d) and NAV‐M (240 mg/d) both significantly reduced attraction, but no significant difference was observed between the two treatment means. NAV‐H (430 mg/d) significantly reduced catches by ～60% and 78% compared to verbenone alone and the baited control, respectively. In a second experiment, combining (–)‐verbenone with NAV (NAVV) increased the effects observed in Experiment 1. NAVV‐M (240 mg/d) resulted in an ～69% and 83% reduction in trap catch compared to verbenone alone and the baited control, respectively. Significantly fewer D. brevicomis were captured in NAVV‐H (430 mg/d) than any other treatment resulting in an ～93% reduction in trap catch compared to the baited control. In a third experiment, NAVV was tested at three release rates for its ability to protect individual ponderosa pines, Pinus ponderosa Dougl. ex Laws., from attack by D. brevicomis. Cumulative release rates varied in direct proportion to tree diameter, but represented quarter, half and full NAVV rates. NAVV significantly reduced the density of D. brevicomis attacks, D. brevicomis successful attacks, and levels of tree mortality on attractant‐baited trees. Only three of 15 NAVV‐treated trees died from D. brevicomis attack while ～93% mortality (14/15) was observed in the untreated, baited control. Quarter and half rates were ineffective for reducing tree mortality.