Dysregulation of the interleukin-17A pathway in endometrial tissue from women with unexplained infertility affects pregnancy outcome following assisted reproductive treatment.

STUDY QUESTION: Which transcriptomic alterations in mid-luteal endometrial scratch biopsies, taken prior to the assisted reproductive treatment (ART) treatment cycle are associated with unsuccessful pregnancy? SUMMARY ANSWER: Dysregulated interleukin-17 (IL-17) pathway components are demonstrated in women who fail to become pregnant after ART. WHAT IS KNOWN ALREADY: Implantation failure is now recognised as a critical factor in unexplained infertility and may be an important component of failed ART. STUDY DESIGN, SIZE, DURATION: Using a prospective longitudinal study design, 29 nulliparous women with unexplained infertility undergoing ART were recruited between October 2016 and February 2018. Mid-luteal stage endometrium and matched serum samples were collected, and patients underwent a single embryo transfer in the subsequent cycle. RNA-seq analysis of endometrial biopsies was performed on the discovery cohort (n = 20). PARTICIPANTS/MATERIALS, SETTING, METHODS: Gene set enrichment analysis of the differentially expressed genes (DEGs) was performed. Endometrium and serum were then prepared for IL-17A analysis by ELISA. MAIN RESULTS AND THE ROLE OF CHANCE: There were 204 differentially expressed protein-coding genes identified in tissue from women who became pregnant (n = 9) compared with tissue from women who failed to become pregnant (n = 11) (false discovery rate; P < 0.05). Of the 204 DEGs, 166 were decreased while 38 were increased in the pregnant compared to the non-pregnant groups. Gene set enrichment analysis of the DEGs identified an over-representation of IL-17 and Pl3K-Akt signalling pathways. All the DEGs within the IL-17 signalling pathway (MMP3, MMP1, IL1ß, LCN2, S100A9 and FOSL1) demonstrated decreased expression in the pregnant group. Serum IL-17 protein levels were increased in the non-pregnant discovery cohort (n = 11) and these findings were confirmed a validation cohort (n = 9). LIMITATIONS, REASONS FOR CAUTION: Limitations of our study include the cohort size and the lack of aneuploidy data for the embryos; however, all embryos transferred were single good or top-quality blastocysts. WIDER IMPLICATIONS OF THE FINDINGS: These findings demonstrate dysregulated IL-17 pathway components in women who fail to become pregnant after ART. Elevated serum levels of the pro-inflammatory cytokine IL-17 may predict failure of ART in women with unexplained infertility. Future trials of anti-IL-17 therapies in this cohort warrant further investigation. STUDY FUNDING/COMPETING INTEREST(S): Funding from the UCD Wellcome Institutional Strategic Support Fund, which was financed jointly by University College Dublin and the SFI-HRB-Wellcome Biomedical Research Partnership (ref 204844/Z/16/Z), is acknowledged. The authors have no competing interests. TRIAL REGISTRATION NUMBER: NA.

Risk factors for human-directed canine aggression in a referral level clinical population.

Risk factors for human-directed aggression were investigated using retrospective analysis of data from a referral-level clinical behaviour population in the UK. A sample of 200 cases involving human-directed canine aggression and 200 control cases involving no instance of human-directed aggression were selected at random from a population of 746 cases. The final model suggested that clinical cases with human-directed aggression were significantly younger than those presenting with other undesired behaviours (P=0.008) and that male dogs were 1.4 times more likely to be aggressive towards human beings than female dogs (P=0.019). Dogs were 1.7 times more likely to be aggressive towards people if they had attended more than five puppy classes than if they had never attended puppy class (P=0.015) and that dogs were 2.8 times more likely to be aggressive towards human beings if there was another dog between 0 months and 24 months of age in the home (P=0.004). These factors only account for 7 per cent to 10 per cent of the variance between the human-directed aggression population and the control population, but factors such as attendance at puppy classes and numbers of dogs in the household suggest the need for longitudinal studies to investigate temporal relationships.

Fertility and genomics: comparison of gene expression in contrasting reproductive tissues of female cattle.

To compare gene expression among bovine tissues, large bovine RNA-seq datasets were used, comprising 280 samples from 10 different bovine tissues (uterine endometrium, granulosa cells, theca cells, cervix, embryos, leucocytes, liver, hypothalamus, pituitary, muscle) and generating 260 Gbases of data. Twin approaches were used: an information-theoretic analysis of the existing annotated transcriptome to identify the most tissue-specific genes and a de-novo transcriptome annotation to evaluate general features of the transcription landscape. Expression was detected for 97% of the Ensembl transcriptome with at least one read in one sample and between 28% and 66% at a level of 10 tags per million (TPM) or greater in individual tissues. Over 95% of genes exhibited some level of tissue-specific gene expression. This was mostly due to different levels of expression in different tissues rather than exclusive expression in a single tissue. Less than 1% of annotated genes exhibited a highly restricted tissue-specific expression profile and approximately 2% exhibited classic housekeeping profiles. In conclusion, it is the combined effects of the variable expression of large numbers of genes (73%-93% of the genome) and the specific expression of a small number of genes (<1% of the transcriptome) that contribute to determining the outcome of the function of individual tissues.

Acute childhood asthma in Galway city from 1985-2005: relationship to air pollution and climate.

We examine the relationship of air pollution and climatic variables to asthma admission rates of children in Galway city over a 21 year period. Paediatric asthma admissions were recorded from 1985-2005, and admission rates per thousand calculated for pre-school (1-4 years), school aged (5-14 years) and all children (1-14 years) on a monthly and annual basis. These data were compared to average monthly and annual climatic variables (rainfall, humidity, sunshine, wind speed and temperature) and black smoke levels for the city. Simple correlation and Poisson Generalized Additive Models (GAM) were used. Admission rates each month are significantly correlated with smoke levels (p = 0.007). Poisson GAM also shows a relationship between admissions and pollution (p = 0.07). Annual smoke levels impact more on admission rates of preschoolers (p = 0.04) than school age children (p = 0.10). These data suggest that air pollution is an important factor in the epidemiology of acute childhood asthma.

Inter-dog aggression in a UK owner survey: prevalence, co-occurrence in different contexts and risk factors.

Aggression between dogs is common and can result in injury. The aims of this study were to estimate prevalence, evaluate co-occurrence with human-directed aggression, and investigate potential risk factors, using a cross-sectional convenience sample of dog owners. Aggression (barking, lunging, growling or biting) towards unfamiliar dogs was reported to currently occur, by 22 per cent of owners, and towards other dogs in the household, by 8 per cent. A low level of concordance between dog and human-directed aggression suggested most dogs were not showing aggression in multiple contexts. Aggression towards other dogs in the household was associated with increasing dog age, use of positive punishment/negative reinforcement training techniques, and attending ring-craft classes. Aggression towards other dogs on walks was associated with location of questionnaire distribution, owner age, age of dog, origin of dog, dog breed type, use of positive punishment/negative reinforcement training techniques and attending obedience classes for more than four weeks. In both, the amount of variance explained by models was low (<15 per cent), suggesting that unmeasured factors mostly accounted for differences between groups. These results suggest general characteristics of dogs and owners which contribute to intraspecific aggression, but also highlight that these are relatively minor predictors.

The incidence and relevance of prostate cancer in radical cystoprostatectomy specimens.

PURPOSE: To review the incidence, histopathological features and clinical outcomes of patients with incidental prostate cancer (CaP) found in cystoprostatectomy specimens (CP) excised for bladder cancer and to determine whether these prostate cancers could affect the follow-up strategy. PATIENTS AND METHODS: We retrospectively reviewed the records of 110 patients who underwent CP for bladder cancer (1998-2011) at our institution. CaP grade, stage, volume and surgical margin status were recorded. Prostatic involvement by bladder tumour or carcinoma in situ (CIS) was studied. Pre-operative prostate assessment and follow-up in those diagnosed with incidental CaP were analysed. RESULTS: Incidental CaP was identified in 35 patients (32.5%), 4 with prostatic PIN alone and 2 patients with diagnoses of CaP prior to cystectomy were excluded from study. Of the CaP cases, 28.5% had clinically significant disease: 5 with Gleason score 7, 2 with Gleason score 9, who also had extracapsular invasion of tumour, and three with positive surgical margins. All patients were pN0 for CaP. Of the 108 patients, 16.5% had prostatic urethral involvement with CIS or TCC. In the subgroup of patients with the incidentally diagnosed CaP who developed local recurrence of bladder tumour and/or metastatic disease, none originated from their CaP. CONCLUSION: The majority of incidental CaP in CP specimens are organ confined and do not influence oncological outcome. The prognosis of such patients is primarily determined by bladder cancer. Our findings support previous reports and autopsy studies elsewhere.

Evidence for an early endometrial response to pregnancy in cattle: both dependent upon and independent of interferon tau.

The aims of this study were to 1) identify the earliest transcriptional response of the bovine endometrium to the presence of the conceptus (using RNAseq), 2) investigate if these genes are regulated by interferon tau (IFNT) in vivo, and 3) determine if they are predictive of the pregnancy status of postpartum dairy cows. RNAseq identified 459 differentially expressed genes (DEGs) between pregnant and cyclic endometria on day 16. Quantitative real-time PCR analysis of selected genes revealed PARP12, ZNFX1, HERC6, IFI16, RNF213, and DDX58 expression increased in pregnant compared with cyclic endometria on day 16 and were directly upregulated by intrauterine infusion of IFNT in vivo for 2 h (P < 0.05). On day 13 following estrous endometrial expression of nine genes increased [ARHGAP1, MGC127874, LIMS2, TBC1D1, FBXL7, C25H16orf71, LOC507810, ZSWIM4, and one novel gene (ENSBTAT00000050193)] and seven genes decreased (SERBP1, SRGAP2, AL7A1, TBK1, F2RL2, MGC128929, and WBSCR17; P < 0.05) in pregnant compared with cyclic heifers. Of these DEGs, significant differences in expression between pregnant and cyclic endometria were maintained on day 16 for F2RL2, LIMS2, LOC507810, MGC127874, TBC1D1, WBSCR17, and ZSWIM4 (P < 0.05) both their expression was not directly regulated by IFNT in vivo. Analysis of the expression of selected interferon-stimulated genes in blood samples from postpartum dairy cows revealed a significant increase (P < 0.05) in expression of ZXFX1, PARP12, SAMD9, and HERC6 on day 18 following artificial insemination in cows subsequently confirmed pregnant compared with cyclic controls. In conclusion, RNAseq identified a number of novel pregnancy-associated genes in the endometrium of cattle during early pregnancy that are not regulated by IFNT in vivo. In addition, a number of genes that are directly regulated by short term exposure to IFNT in vivo are differentially expressed on day 18 following estrus detection in the blood of postpartum dairy cows depending on their pregnancy status.

Effect of the metabolic environment at key stages of follicle development in cattle: focus on steroid biosynthesis.

Cellular mechanisms that contribute to low estradiol concentrations produced by the preovulatory ovarian follicle in cattle with a compromised metabolic status are largely unknown. To gain insight into the main metabolic mechanisms affecting preovulatory follicle function, two different animal models were used. Experiment 1 compared Holstein-Friesian nonlactating heifers (n = 17) and lactating cows (n = 16) at three stages of preovulatory follicle development: 1) newly selected dominant follicle in the luteal phase (Selection), 2) follicular phase before the LH surge (Differentiation), and 3) preovulatory phase after the LH surge (Luteinization). Experiment 2 compared newly selected dominant follicles in the luteal phase in beef heifers fed a diet of 1.2 times maintenance (M, n = 8) or 0.4 M (n = 11). Lactating cows and 0.4 M beef heifers had higher concentrations of ß-hydroxybutyrate, and lower concentrations of glucose, insulin, and IGF-I compared with dairy heifers and 1.2 M beef heifers, respectively. In lactating cows this altered metabolic environment was associated with reduced dominant follicle estradiol and progesterone synthesis during Differentiation and Luteinization, respectively, and in 0.4 M beef heifers with reduced dominant follicle estradiol synthesis. Using a combination of RNA sequencing, Ingenuity Pathway Analysis, and qRT-PCR validation, we identified several important molecular markers involved in steroid biosynthesis, such as the expression of steroidogenic acute regulatory protein (STAR) within developing dominant follicles, to be downregulated by the catabolic state. Based on this, we propose that the adverse metabolic environment caused by lactation or nutritional restriction decreases preovulatory follicle function mainly by affecting cholesterol transport into the mitochondria to initiate steroidogenesis.

Autologous attenuated T-cell vaccine (Tovaxin) dose escalation in multiple sclerosis relapsing-remitting and secondary progressive patients nonresponsive to approved immunomodulatory therapies.

An open-label dose escalation study of T-cell vaccination in multiple sclerosis patients was conducted using attenuated myelin reactive T-cells (MRTC) selected with six myelin peptides, two each from MBP, PLP and MOG. The dose range of subcutaneous injections given at weeks 0, 4, 12 and 20 was 6-9E6, 30-45E6 and 60-90E6 irradiated MRTC. Assessments were over 52 weeks for MRTC levels, EDSS, MSIS-29, brain MRI and relapses. The 30-45E6 dose was the most effective with reductions in MRTC ranging from 92.4% at week 5 to 64.8% at week 52. The reduction in relapses compared to baseline for the M-ITT and evaluable per-protocol analyses were 63.5%, and 85.0% at week 52. The MRI lesions were stable while there was an improvement trend in the EDSS and MSIS-29 physical subscore following the second injection. Adverse events were mild to moderate in intensity with mild injection site reactions occurring with increasing dosage. The mid-dose was selected for further clinical development studies because of the rapid depletion of peripheral blood MRTC and a trend for improvements in clinical outcomes following immunization.

Subacute cutaneous lupus erythematosus presenting as poikiloderma.

Subacute cutaneous lupus erythematosus (SCLE) is a recognised variant of lupus erythematosus (LE), which accounts for 10-15% of all cases of cutaneous LE, occurring most commonly in young to middle-aged white women. Diagnosis is based on the detection of anti-Ro/SS-A antibodies in the skin and serum, characteristic clinical and histological cutaneous involvement, and relatively mild systemic involvement. Several unusual variants of SCLE have been reported including erythrodermic SCLE, SCLE with vitiligo-like lesions, acral SCLE and bullous SCLE. Poikoilodermatous SCLE is a recognised but rare variant of SCLE. There are currently only two case reports, comprising five individual cases, in the literature. We present a case of SCLE in which the main clinical findings were an extensive photodistributed poikilodermatous rash and alopecia.

Services for children with diabetes.

This is the first assessment of the services available and care received by children and adolescents with Type 1 Diabetes in the Republic of Ireland. Questionnaires were mailed to 19 centres in 2006 identified as providing care for children and adolescents with Diabetes. Replies were received from all centres. 29 consultants provide care for a reported 2040 patients in the 19 centres. 16 (50%) paediatricians surveyed expressed a special interest in Diabetes while 27 (94%) see children in a designated Diabetes clinic. The average caseload per whole-time-equivalent (WTE) Diabetes Nurse Specialists (DNS) is 162 patients and 416 patients per WTE Dietician. 13 of the 19 centres (70%) have no access to Psychology services. This review of children's diabetes services highlights the current deficiencies, particularly in relation to support staff, with respect to international recommendations. Improvements in resources to reduce long-term complications needs urgent addressing at both local and national levels.

Self-reported pre-pregnancy maternal body mass index and infant birth-weight.

This analysis examines the association between maternal characteristics, particularly body mass index (BMI) and infant birth weight in 1048 live infants. Mean reported pre pregnancy BMI of mothers was 23.74 kg/m2 (SD 4.21). The educational level of the mother's parents was independently associated with maternal BMI, those with higher educated parents having a lower reported BMI (F = 2.787, p = 0.029). Mean infant birth weight was 3493 g (SD 18.1) and there was a strong graduated relationship to estimated gestational age. In a sub-group of participating maternal grandmothers (n = 171), reported BMI was 26.7Kg/m2. The BMI of expectant mothers was significantly associated with their own mother's BMI. (r = 0.179, p = 0.005) in this sub-group. These preliminary findings, which will be investigated further with recorded height and weight information, suggest that familial factors are influential, perhaps through genetic predisposition or shared socio-cultural factors such as diet.

Mothers' dietary patterns during pregnancy and risk of asthma symptoms in children at 3 years.

There is increasing evidence that dietary factors in early life play a role in the aetiology of childhood asthma. Our objective in this analysis was to assess whether maternal dietary patterns during pregnancy, as measured by a validated food frequency questionnaire, influenced general practitioner diagnosis of asthma by the age of 3 years in the Life-ways cohort of children. General Practice follow-up records were available for 631 of 1001 singleton children, twins having been excluded (63% follow-up rate). Overall 10.4% of children had diagnosed asthma, a prevalence rate comparable with other studies. In logistic regression models, based on quartiles of intake, which adjusted for maternal lifestyle and socio-economic circumstances, relatively higher maternal fruit and vegetable intake and oily fish consumption were associated with lower risk of children developing asthma, whilst those with relatively higher spreadable fat intake had a higher risk of asthma. These findings warrant further investigation as they imply an important role for maternal diet in childhood asthma, though the confounding effect of other social and lifestyle factors should be assessed as the children get older.

Prevalence of asthma in Galway school children 2004.

We aimed to ascertain the prevalence of asthma amongst Galway schoolchildren aged 9-10, and to compare the results to a similar study carried out in 1992. A questionnaire was distributed to parents of all children attending 4th class in Galway city schools. 652 of 750(87%) questionnaires were returned. Prevalence of "wheeze ever" was 27.6%, and "current wheeze" 16.3%. The prevalence of asthma was 18.5%. Of those with asthma there was a 2 to 1 male preponderance, 80% had mild asthma, 14% moderate, and 6% severe asthma. 80% were taking anti-asthma therapy, with 48% taking regular inhaled steroids. 84% had a diagnosis of asthma made by a doctor. Comparison with the study of 1992 shows little change in the prevalence of current wheeze, or asthma. There has however been a significant decline in the severity of asthma, and an increase in the use of prophylactic anti-asthma medication. Asthma prevalence appears to be stable in the age group studied. There is a much greater willingness to diagnose, and treat asthma in the community. The severity of asthma, as measured by attack frequency, has declined.

Patterns of asthma admissions in children.

Admissions for childhood asthma have declined over the past 10 years in the U.K. The reasons for this decline are not known. There are few published data on Irish children. The study was completed in a paediatric unit in a regional general hospital, with a stable urban/rural catchment population of approximately 45,000 children under 15 years. H.I.P.E. data on paediatric asthma admissions were analysed from 1990-2004. A prospective study of all acute asthma admissions of children aged 1-14 years over a 12 -month period was also undertaken, data were compared to a similar study in 1990, and a retrospective study in 1995. The number of admissions rose to a peak in 1995, and there was a large decline in the number of admissions thereafter. The fall was most marked in school age children. The prospective study showed that the treatment of asthma in the community, and in hospital was more aggressive. There were more children diagnosed, and on prophylactic therapy prior to admission. There was greater use of oxygen and nebulised ipratropium in hospital; milder cases were discharged sooner. Subjective assessment of severity of attacks on admission indicated increased severity, but objective measures showed less change. We have shown that the fall in asthma admissions was accompanied by more aggressive management in the community, with little changes in the attack severity in those admitted. The reduction in admissions was most evident in school age children. These data are consistent with the hypothesis that better management of asthma in the community is responsible for the reduction in admissions.

In silico mining identifies IGFBP3 as a novel target of methylation in prostate cancer.

Promoter hypermethylation is central in deregulating gene expression in cancer. Identification of novel methylation targets in specific cancers provides a basis for their use as biomarkers of disease occurrence and progression. We developed an in silico strategy to globally identify potential targets of promoter hypermethylation in prostate cancer by screening for 5' CpG islands in 631 genes that were reported as downregulated in prostate cancer. A virtual archive of 338 potential targets of methylation was produced. One candidate, IGFBP3, was selected for investigation, along with glutathione-S-transferase pi (GSTP1), a well-known methylation target in prostate cancer. Methylation of IGFBP3 was detected by quantitative methylation-specific PCR in 49/79 primary prostate adenocarcinoma and 7/14 adjacent preinvasive high-grade prostatic intraepithelial neoplasia, but in only 5/37 benign prostatic hyperplasia (P < 0.0001) and in 0/39 histologically normal adjacent prostate tissue, which implies that methylation of IGFBP3 may be involved in the early stages of prostate cancer development. Hypermethylation of IGFBP3 was only detected in samples that also demonstrated methylation of GSTP1 and was also correlated with Gleason score > or =7 (P=0.01), indicating that it has potential as a prognostic marker. In addition, pharmacological demethylation induced strong expression of IGFBP3 in LNCaP prostate cancer cells. Our concept of a methylation candidate gene bank was successful in identifying a novel target of frequent hypermethylation in early-stage prostate cancer. Evaluation of further relevant genes could contribute towards a methylation signature of this disease.

Access of asylum seeker children to acute paediatric services.

To investigate the interface between primary care and paediatric services in the referral of asylum seekers. Over a 3 month period a questionnaire was administered, and clinical data gathered on every child attending the A&E department of UCHG whose parents were seeking asylum in this country. Control data was obtained for the next Irish child seen on-call. At the time of presentation to the paediatric service, an Irish child was 4 times more likely (32%) to have initially been seen and referred by a GP than an asylum seeker child (8%); 80% of asylum seeker families had registered with a GP, compared to 96% of controls. 24% of asylum seeker families had called and used an emergency response ambulance to get to hospital, compared to just 4% of Irish children. The rate of subsequent admission to the paediatric ward from A&E was nearly that in asylum seeker children (24%) compared to Irish controls (40%), get to hospital, compared to just 4% of Irish children. Asylum seeker children are less likely to have seen a GP prior to A&E presentation, more likely to go to hospital by ambulance and less likely to be subsequently admitted, suggesting an over-dependence on paediatric hospital services in this population.

Heterogeneous expression of alpha-methylacyl-CoA racemase in prostatic cancer correlates with Gleason score.

AIMS: Alpha-methylacyl-CoA racemase (AMACR) is a sensitive and specific immunohistochemical marker of prostatic malignancy, staining 80-100% of prostatic cancers with absent staining in benign glands. However, positive staining in benign conditions as well as low rates of AMACR reactivity in prostatic cancer variants have been described. Preliminary use of AMACR immunohistochemistry in our institution has suggested lower specificity and sensitivity for prostatic cancer than initially proposed. The aim of this study was to establish true rates of AMACR reactivity in prostatic cancer and benign prostatic hyperplasia (BPH). METHODS AND RESULTS: AMACR immunohistochemistry was performed on sections from 57 prostatic cancers and 44 BPH resections. Ninety-one percent of cancers were AMACR+, with diffuse (> 75%) tumour staining in 53% of cases. Thirty-eight percent of tumours showed heterogeneous expression (1-75% tumour staining). This was significantly correlated with increased Gleason score. High-grade prostatic intraepithelial neoplasia (PIN) was AMACR+ in 87% of cancers. Eleven percent of BPH showed moderate or strong staining in benign glands, focally mimicking the malignant staining pattern. CONCLUSIONS: This study confirms heterogeneous AMACR expression in prostatic cancer and shows a correlation with Gleason score. Positive staining in BPH is also documented, thus emphasizing the importance of interpreting AMACR immunohistochemistry in the context of other findings in a diagnostic setting.

Rapid assessment breast clinics--evolution through audit.

This observational, cohort study aimed to examine the potential utility of Rapid Assessment Breast Clinics (RABC) beyond cancer detection at presentation. One thousand four hundred and twenty nine women were studied over an 18 month period. 154 (10.7%) had breast cancer - 87.7% of whom were seen expediently with 92.9% being diagnosed at one attendance. One hundred and forty three (10%) of those with a benign diagnosis were found by routine questioning to have significant familial risk separate to their reason for referral. Despite careful triage, considerable contamination of appointment allotment occurred with many who were correctly triaged as non-urgent being seen 'urgently'. One hundred and seventy six attendees (12.3%) had neither the symptom that triggered referral, nor breast lump, nipple discharge nor family history of breast cancer, while 283 (19.8%) had no objective clinical or radiological abnormality. Although RABC reliably categorise malignant versus non-malignant diagnoses despite cluttering by low risk women, a significant proportion of non-cancer patients still require address of future risk rather than reassurance of their present status alone.