Functional and Behavioral Trade-Offs between Terrestrial and Aquatic Locomotion in the Amphibious Fish Kryptolebias marmoratus.

The mangrove rivulus (Kryptolebias marmoratus) is a phenotypically plastic teleost fish that can spend considerable time on land and traverse the terrestrial realm through a behavior termed the tail-flip jump. The tail-flip jump is a transitional stage between fully aquatic and terrestrial lifestyles. Therefore, understanding this behavior can provide insight into how organisms adapt to new environments over evolutionary time. Studies of K. marmoratus show that terrestrial acclimation and exercise improve tail-flip jumping performance due to muscle remodeling, but the implications of these muscular changes on aquatic locomotion are unknown. In the present study, we hypothesized that (1) terrestrial acclimation and exercise lead to physiological changes, such as changes to muscle fiber type, muscle mass distribution, or body shape, that optimize tail-flip jump distance and endurance while negatively impacting swimming performance in K. marmoratus, and (2) plasticity of the brain (which has been demonstrated in response to a variety of stimuli in K. marmoratus) allows terrestrial emersion and exercise to cause behavioral changes that promote survival and long-term reproductive success. To test these hypotheses, we measured the critical swimming speed (Ucrit), tail-flip jump distance, terrestrial endurance, and undisturbed aquatic behavior of age- and size-matched K. marmoratus before and after a terrestrial exercise period. This period consisted of six 3-min exercise sessions spread over 12 days, during which the fish were prompted to jump continuously. To isolate the effects of air exposure, a separate group was exposed to air for an equivalent period but not allowed to jump. Air exposure improved maximum jump distance but negatively affected swimming performance (Ucrit). Terrestrial endurance (number of jumps) improved in the exercised group, but Ucrit showed no significant change. Contrary to our first hypothesis, a trade-off exists between jump distance and Ucrit but not between jump endurance and Ucrit. Exercised individuals were more active following exercise, resulting either from the onset of dispersion behavior or a heightened stress response.

Histological Features and Tissue Microarray Taxonomy of Nigerian Breast Cancer Reveal Predominance of the High-Grade Triple-Negative Phenotype.

INTRODUCTION: Little is known about the biology, molecular profile and hence optimal treatment of African Nigerian breast cancer. The aim of this work, therefore, was to characterize the histology and molecular profile of Nigerian breast cancer. METHODS: Breast carcinomas from women at 6 centres of similar tribal origin in Nigeria were reviewed and assembled into tissue microarrays (TMAs), and sections were stained for hormone receptors, i.e. estrogen receptor (ER)α, ERß1, ERß progesterone receptor (PR) and androgen receptor, cyclin D, HER2, Ki67 and cytokeratins (CKs), i.e. CK5/6 and CK14 (basal) and CK18 and 19 (luminal). RESULTS: A total of 835 tumours were analysed. The mean age at diagnosis was 48.62 ± 12.41 years. The most common histological subtype was ductal NST (no-special-type) carcinoma (87.3%). Over 90% of the tumours were grade 2 or 3. The predominant molecular phenotype was the non-basal, triple-negative type (47.65%) followed by the HER2-positive group (19.6%). The percentage of ER-, PR- and HER2-positive tumours was 22.4, 18.9 and 18.8%, respectively. CONCLUSION: Nigerian breast cancer predominantly has a high-grade, triple-negative profile. It occurs at a younger age and bears similarities at the molecular level to pre-menopausal breast cancer in white women, with remarkably lower levels of ERß expression. The early presentation and histological and molecular phenotype may explain the poor prognosis, and tailoring treatment strategies to target this unique profile are required.

Detection of Lewis antigen structural change by FTIR spectroscopy.

Mucins are a family of extensively glycosylated, high molecular weight glycoproteins. Secretion of mucins with altered terminal carbohydrate moieties alters the rheological and viscoelastic properties of mucus and observed glycosylation changes in respiratory diseases may vary with disease status. Structural modifications to the Lewis x antigen with sialic acid (sialyl-Lewis x) and sulphate (sulfo-Lewis x) in particular are associated with respiratory diseases and deemed potential biomarkers for disease diagnosis, severity and progression. The major aim of this study was to evaluate the ability of Fourier transform infrared spectroscopy (FTIR) to detect, via infrared (IR) spectra, the structural changes between the Lewis x antigen and sialylated and sulphated derivatives. Although FTIR only provides information on vibrations of chemical groups, we show that by comparing mono- and oligosaccharide specific IR spectra it is possible to determine the contribution of key sugar moieties to the altered Lewis x spectral pattern.

Loss of WT1 expression in the endometrium of infertile PCOS patients: a hyperandrogenic effect?

CONTEXT: In fertile patients the endometrial Wilms tumor suppressor gene (WT1) is expressed during the window of implantation. Polycystic ovary syndrome (PCOS) patients suffer from hyperandrogenemia and infertility and have elevated endometrial androgen receptor (AR) expression. WT1 is known to be down-regulated by AR. Therefore, the expression of WT1 and its targets may be altered in PCOS endometrium. OBJECTIVE: The objective of the study was to assess the expression and regulation of WT1 and selected downstream targets in secretory endometrium from ovulatory PCOS (ovPCOS) and fertile women. DESIGN AND PATIENTS: Endometrial samples were obtained from 25 ovPCOS and 25 fertile patients. MAIN OUTCOME MEASURE: Endometrial expression of WT1 and selected downstream targets were assessed by immunohistochemistry and RT-PCR. The androgen effect on WT1 expression was determined in vitro by immunoblots and RT-PCR. The expression of WT1 and its targets was quantified in fertile and ovPCOS stromal cells in the presence of androgens by RT-PCR. Caspase-3/7 activity was measured to evaluate sensitivity to drug-induced apoptosis. RESULTS: WT1 expression was down-regulated in secretory-phase ovPCOS endometrium. Stromal expression of Bcl-2 and p27 was higher, and epidermal growth factor receptor was lower in ovPCOS than in fertile patients. Endometrial stromal expression of WT1, Bcl-2, Bcl-2-associated X protein, and ß-catenin was regulated by androgens. Apoptosis levels were reduced in ovPCOS samples and androgen-treated fertile samples. CONCLUSION: WT1 expression is down-regulated in ovPCOS endometrium during the window of implantation. Androgens regulate the expression of WT1 and its targets during endometrial decidualization. The altered balance between WT1 and AR in the endometrium of PCOS patients may jeopardize the success of decidualization and endometrial receptivity.

Reproductive status in broiler breeder males is minimally affected by a mid-cycle increase in photoperiod.

1. Broiler breeder males were reared on 8-h photoperiods, transferred to 12 h at 20 weeks, and either maintained on 12 h or further transferred to 14, 16, 18, 20 or 22 h at 40 weeks to determine whether reproductive status and the timing of the onset of adult photorefractoriness is influenced by photoperiod during the breeding cycle. 2. No photoperiodic effect was observed on sperm concentration although the combination of large variation within a treatment and the lack of a semen sample from some males at each collection may have masked small differences and indicates that perhaps fertility in individual males fluctuates naturally during the breeding season. 3. The rate of comb growth to, and testicular weights at, 60 weeks were not significantly different between treatments. 4. Plasma testosterone concentration at 60 weeks of age declined linearly with photoperiod between 12 and 22 h, suggesting that in the birds exposed to the more stimulatory photoperiods after 40 weeks, hypothalamic-pituitary-gonadal activity shut down earlier. 5. An increase in photoperiod at 40 weeks of age appears to advance the onset of photorefractoriness in male broiler breeders, evident from the decline in testosterone concentration with increasing photoperiod; it is however insufficient to affect sperm concentration or comb area.

Temporary exposure of prepubertal, somatically-mature broiler breeder females to stimulatory photoperiods.

1. Individually-caged Ross 308 broiler breeders were transferred temporarily at 147 d from 8 to 14-h photoperiods and back to 8 h at 5-d intervals from 5 to 25 d and at 32 d. Control birds were maintained on 8 h or transferred permanently to 14 h. 2. Five long days had minimal effect on age at first egg, between 10 and 25 long days progressively accelerated sexual development, and maximum advance was achieved by 32 long days. However, a model involving the ogive of a normal distribution with a mean of 14 long days and an SD of 5.3 indicated that 28 long days would be sufficient to maximise sexual development. 3. The similarity of this model for broiler breeders with one previously produced for egg-type pullets suggested a common physiological mechanism for photosexual responses to two opposing changes in photoperiod.

Somatic sex identity is cell autonomous in the chicken.

In the mammalian model of sex determination, embryos are considered to be sexually indifferent until the transient action of a sex-determining gene initiates gonadal differentiation. Although this model is thought to apply to all vertebrates, this has yet to be established. Here we have examined three lateral gynandromorph chickens (a rare, naturally occurring phenomenon in which one side of the animal appears male and the other female) to investigate the sex-determining mechanism in birds. These studies demonstrated that gynandromorph birds are genuine male:female chimaeras, and indicated that male and female avian somatic cells may have an inherent sex identity. To test this hypothesis, we transplanted presumptive mesoderm between embryos of reciprocal sexes to generate embryos containing male:female chimaeric gonads. In contrast to the outcome for mammalian mixed-sex chimaeras, in chicken mixed-sex chimaeras the donor cells were excluded from the functional structures of the host gonad. In an example where female tissue was transplanted into a male host, donor cells contributing to the developing testis retained a female identity and expressed a marker of female function. Our study demonstrates that avian somatic cells possess an inherent sex identity and that, in birds, sexual differentiation is substantively cell autonomous.

Photoperiods for broiler breeder females during the laying period.

Ross 308 broiler breeder females, maintained in litter-floor pens, were reared from 2 d of age on 8-h photoperiods and transferred abruptly to an 11-, 12-, 13-, or 14-h photoperiod at 20 wk of age and at a mean BW of 2.09 kg. Four qualitative dietary treatments were superimposed upon each photoperiodic treatment, but, in the absence of any significant interactions of photoperiod with diet, only photoperiodic means are reported. Age at 50% egg production (ASM) was progressively advanced as photoperiod increased from 11 to 13 h but was similar for 13-h and 14-h birds. Egg production was significantly related to ASM, with peak egg numbers being stimulated by a transfer to 13-h photoperiods. Rate of lay after 48 wk of age declined more rapidly in the 14-h birds than in any of the other groups, suggestive of an earlier onset of adult photorefractoriness. More eggs were laid outside the nest box and more cracked and dirty eggs were produced by birds on 11- and 12-h photoperiods than on 13- or 14-h photoperiods. Mean egg weight was positively correlated with ASM but negatively related to photoperiod; the number of eggs thought to be double-yolked was not significantly affected by photoperiod. Total egg mass output was not significantly affected by photoperiod. Birds exposed to 11-h and 12-h photoperiods were heavier at 60 wk than birds transferred to 13- or 14-h photoperiods. These findings show that a transfer from 8 to 13 h at 20 wk optimizes egg-laying performance in broiler breeder females, especially when they are kept beyond 60 wk of age.

Photoperiodic responses of broilers. I. Growth, feeding behaviour, breast meat yield, and testicular growth.

1. A total of 7960 Cobb and Ross broiler males were reared in two trials to 35 d on various photoperiods between 2 and 21 h or under continuous illumination; a total of 444 birds were randomly selected at 35 d and retained for subsequent determination of breast meat yield and testicular weight at 40 or 54 d of age. 2. In both strains, feed intake and growth were positively correlated with photoperiod during the first 21 d, but afterwards feed intake was not significantly affected by photoperiods longer than 6 h and growth was negatively correlated with photoperiod beyond 12 h. Overall, to 35 d, there were no significant photoperiodic influences on either feed intake or growth for >or=6-h photoperiods, but significant depressions in feed intake and growth for photoperiods shorter than 6 h. Feed conversion efficiency was maximised by 12-h photoperiods, with decreases in efficiency above and below 12 h. Mortality was unaffected by photoperiod <12 h, but increased proportionately with photoperiod >12 h. The incidence of Sudden Death Syndrome (SDS) had an inverse relationship with photoperiod 10 h. The European Efficiency Factor was curvilinearly related to photoperiod, with the highest efficiency occurring at 12 h. Ross birds had significantly greater feed intakes but poorer feed conversion efficiencies than Cobb; differences in growth, overall mortality and the incidence of SDS between the strains were not significantly different. 3. By 5 d, birds given

Photoperiodic responses of broilers. II. Ocular development.

1. Poultry are naturally long-sighted when they hatch, becoming emmetropic by 6 weeks of age. However, Cobb and Ross broilers exposed to

Photoperiodic responses of broilers. III. Tibial breaking strength and ash content.

1. A total of 7960 Cobb and Ross broiler males were reared on various photoperiods or continuous illumination in two trials to 35 or 40 d. Tibial breaking strength was measured in both, and tibial ash content determined in the first of the two trials. 2. Tibial breaking strength was significantly affected by photoperiod, body weight, testicular weight, and genotype. Although peak bone strength occurred at about 7 h for Ross and at 12 h for Cobb, with reductions in strength for both shorter and longer photoperiods than these peaks, the removal of body weight effects showed that tibial breaking strength was negatively correlated with photoperiod per se. However, after removal of photoperiodic influences, breaking strength was positively correlated with both body weight and testicular weight. 3. Ross birds had greater tibial breaking strengths than Cobb, irrespective of whether the comparisons were made at the same photoperiod, the same body weight, or the same ash content. 4. Ash content increased according to the logarithm of photoperiod, but was unrelated to body weight. Tibial breaking strength increased with ash content. Ash contents were similar for Ross and Cobb birds. 5. Despite the significant differences in breaking strength and ash content, there were no broken bones or signs of leg abnormality in any of the 300 birds sampled. Tibial breaking strength data may therefore be of limited value in assessing the benefits of photoperiod to welfare in modern broilers.

A study of mitochondrial DNA D-loop mutations and p53 status in nonmelanoma skin cancer.

BACKGROUND: Mitochondrial DNA (mtDNA) displacement-loop (D-loop) mutations have previously demonstrated potential as smoking-induced biomarkers in oral squamous cell carcinoma (SCC). Additionally, they have been observed in SCC and basal cell carcinoma of nonmelanoma skin cancer (NMSC). However, they have not been examined in the SCC precursor lesions, Bowen disease or actinic keratosis. OBJECTIVES: Here, we present a novel study of mtDNA D-loop mutations in these two precursors, a rare keratoacanthoma and NMSC (all tumours not related to smoking). METHODS: We used a polymerase chain reaction and direct sequencing approach. Furthermore, as the tumour suppressor protein p53 has been reported as having a novel role in maintaining mitochondrial genetic stability, we assessed p53 status using immunohistochemistry, evaluating potential association with the presence of mtDNA mutations. RESULTS: Of 36 tumours, nine (25%) exhibited mutations in the D-loop. In total, 13 base substitutions were observed across all patients: seven (53.8%) were A : T to G : C; two (15.4%) were G : C to T : A; two (15.4%) were G : C to A : T and two (15.4%) were G : C to C : G. Four of the 13 (30.8%) base substitutions were observed at nucleotide 146. We observed abnormal p53 accumulation in over half of the samples analysed (55.5%), suggesting it to be a major part of the carcinogenic process of NMSC; however; there was no association between p53 positivity and the presence of mtDNA mutations (P = 0.47). CONCLUSIONS: It is unlikely that alteration in p53 status is a contributing factor to mtDNA mutagenesis.

L-selectin ligands in human endometrium: comparison of fertile and infertile subjects.

BACKGROUND: L-selectin ligands, localized to the luminal epithelium at the time of implantation, may support the early stages of blastocyst attachment. We have assessed the expression of two L-selectin ligands, defined by MECA-79 and HECA-452 monoclonal antibodies, and the sulfotransferase GlcNAc6ST-2, involved in generation of L-selectin ligand epitopes, in the secretory phase of the endometrium from fertile and infertile patients. METHODS: Endometrial samples were obtained from 33 fertile, 26 PCOS, 25 endometriosis and 33 patients diagnosed with unexplained infertility. L-selectin ligands and GlcNAc6ST-2 expression was assessed by immunohistochemistry and immunoblotting. RESULTS: Immunohistochemical staining of uterine epithelium, from fertile and infertile women, demonstrated differential expression of MECA-79 and HECA-452 epitopes. In fertile women in the secretory phase MECA-79 was more strongly expressed, particularly on the lumen, than in infertile women. HECA-452 staining was significantly stronger in the glands in PCOS and endometriosis patients than in fertile women. GlcNAc6ST-2 expression was reduced in infertile patients, correlating with MECA-79 expression. CONCLUSIONS: This study demonstrated significant differences in expression of L-selectin ligands between fertile and infertile women in natural cycles, and could contribute to patient assessment prior to initiating fertility treatment.

Illuminance and egg production in broiler breeders.

1. Ross broiler breeders were reared at a nominal illuminance of 15, 20 or 45 lux and transferred to a nominal illuminance of 25, 55 or 70 lux at 20 weeks. 2. There were no significant interactions between the response to illuminance during rearing and in lay. This means that it matters not whether illuminance is increased, decreased or held constant on transfer to the laying house, provided it equals or exceeds the biological optimum for satisfactory egg production. 3. Whilst there were no significant effects of illuminance in either the rearing period or laying periods on egg numbers, peak rate of lay, terminal rate of lay, egg mass output or liveability, meta-analyses of these and other data indicated biological optima of 15 lux during rearing and 7 lux in the laying period. Birds reared at 45 lux had a lower mean egg weight (and earlier sexual maturity) than birds reared at 15 lux, and hens illuminated at 25 lux in the laying period laid more eggs on the floor than at either 55 or 70 lux. 4. Typical primary breeder recommendations of 10-20 lux during rearing and 30-60 lux in lay are appropriate for floor-housed birds; however, an illuminance of 7 lux could be used for caged birds, subject to welfare-code compliance.

Illuminance, sexual maturation, and early egg production in female broiler breeders.

1. Broiler breeders were reared at an initial illuminance of 13, 21 or 44 lux and transferred at 20 weeks and a mean body weight of 2.06 kg to floor pens at 25, 55 or 71 lux, or to individual cages at various intensities between 3 and 138 lux. 2. Rate of sexual maturation was not significantly affected by the light intensity experienced during the rearing period, but was progressively accelerated by the provision of a brighter illuminance after 20 weeks, up to a ceiling of about 14 lux. There was no interaction between the pre- and post-20-week illuminance. 3. Cumulative egg numbers to 39 weeks were unaffected by illuminance in the rearing period, positively correlated with illuminance post 20 weeks up to an asymptote at about 8 lux, and highly correlated with age at sexual maturity. Peak rate of lay was both delayed and depressed at 3 lux. 4. It is concluded that the minimum light intensity required at bird-head height to achieve satisfactory photostimulation and an optimal peak rate of lay in broiler breeders is 10 lux, but that a higher illuminance would be prudent for broiler breeders kept in floor pen facilities to maximise nest-box usage.

Photoperiodic response curves for plasma LH concentrations and age at first egg in female broiler breeders.

The objective of this work was to define precisely the response curve for photoinduced luteinizing hormone (LH) release in feed-restricted meat-type (broiler) breeder females and to compare it with the photoperiodic response curve for advance in age at first egg (AFE). Birds with a mean body weight of 2.0kg at 20 weeks of age were transferred from an 8 to a 9, 9.5, 10, 10.5, 11, 11.5, 12, 12.5, 13, 14 or 18-h photoperiod; change in plasma LH was measured 4d after photostimulation and subsequent individual AFE recorded. The first significant increase in LH secretion was seen in birds transferred to an 11.5-h photoperiod, but no further significant increases in LH were observed in birds transferred to longer photoperiods. A photoperiodic response curve based on a meta-analysis of changes in photoinduced LH secretion observed in this study and data from an earlier experiment using dwarf broiler breeders indicated a critical daylength of about 9.5h and a saturation daylength of approximately 13h. Similarly, the first significant advance in AFE occurred in birds transferred to an 11-h photoperiod, but with no further significant increases seen in birds transferred to photoperiods >11h. A response curve for photoinduced advances in AFE was produced by meta-analysis using data from the present study and from an earlier investigation involving fewer, more widely spaced photoperiods. It is concluded, in female broiler breeders, that the photoperiodic response curves for photoinduced LH release and AFE are similar, with the point at which the responses begin to rise steeply (classical critical daylength) occurring at 9.5h and the asymptote (classical saturation daylength) at 13h. Functionally, however, the minimum photoperiod to achieve a significant change in either LH secretion or advance in AFE is between 11 and 11.5h.

Comparison of induced and cancer-associated mutational spectra using multivariate data analysis.

One of the most useful tools for investigating the aetiopathology of cancer is the mutation spectrum, which comprises the type and distribution of mutations within a gene sequence. Many studies have generated mutagen-induced spectra using in vitro or in vivo model systems in an attempt to find correlations with those observed in cancer-associated genes such as the TP53 tumour suppressor gene. Consequently, meaningful similarities in the types of mutation found in induced and human spectra have been demonstrated. However, it is more difficult to draw such conclusions about the distribution or sequence context of mutations when they arise in different target sequences. We have developed an analytical approach for base substitution spectra that capture information for both sequence context and mutation type simultaneously. The resulting mutation signature is a fixed set of data points that allows comparison of multiple mutation spectra regardless of sequence. We have applied this method to a mixed set of mutation spectra observed in exons 5, 7 and 8 of TP53 from cancers of brain, breast, skin, colon, oesophagus, liver, head and neck, stomach and lung (smokers and non-smokers) and spectra induced by benzo[a]pyrene diol epoxide, ultraviolet (UV) B, UVC, simulated sunlight and hydroxyl radicals in the cII, supF and yeast p53 model systems. We demonstrate that this approach allows human cancer and mutagen-induced signatures to be grouped together according to similarity. Specifically, the analysis reveals key differences between smoking- and non-smoking-related lung cancer for TP53 mutations and the mutability of CpG sites between exons in skin cancer.