Ectopic Expression of Xylella fastidiosa rpfF Conferring Production of Diffusible Signal Factor in Transgenic Tobacco and Citrus Alters Pathogen Behavior and Reduces Disease Severity.

The pathogenicity of Xylella fastidiosa is associated with its ability to colonize the xylem of host plants. Expression of genes contributing to xylem colonization are suppressed, while those necessary for insect vector acquisition are increased with increasing concentrations of diffusible signal factor (DSF), whose production is dependent on RpfF. We previously demonstrated that transgenic citrus plants ectopically expressing rpfF from a citrus strain of X. fastidiosa subsp. pauca exhibited less susceptibility to Xanthomonas citri subsp. citri, another pathogen whose virulence is modulated by DSF accumulation. Here, we demonstrate that ectopic expression of rpfF in both transgenic tobacco and sweet orange also confers a reduction in disease severity incited by X. fastidiosa and reduces its colonization of those plants. Decreased disease severity in the transgenic plants was generally associated with increased expression of genes conferring adhesiveness to the pathogen and decreased expression of genes necessary for active motility, accounting for the reduced population sizes achieved in the plants, apparently by limiting pathogen dispersal through the plant. Plant-derived DSF signal molecules in a host plant can, therefore, be exploited to interfere with more than one pathogen whose virulence is controlled by DSF signaling.

Expression of Xylella fastidiosa RpfF in citrus disrupts signaling in Xanthomonas citri subsp. citri and thereby its virulence.

Xylella fastidiosa and Xanthomonas citri subsp. citri, that cause citrus variegated chlorosis (CVC) and citrus canker diseases, respectively, utilize diffusible signal factor (DSF) for quorum sensing. DSF, produced by RpfF, are similar fatty acids in both organisms, although a different set of genes is regulated by DSF in each species. Because of this similarity, Xylella fastidiosa DSF might be recognized and affect the biology of Xanthomonas citri. Therefore, transgenic Citrus sinensis and Carrizo citrange plants overexpressing the Xylella fastidiosa rpfF were inoculated with Xanthomonas citri and changes in symptoms of citrus canker were observed. X. citri biofilms formed only at wound sites on transgenic leaves and were thicker; however, bacteria were unable to break through the tissue and form pustules elsewhere. Although abundant growth of X. citri occurred at wound sites on inoculated transgenic leaves, little growth was observed on unwounded tissue. Genes in the DFS-responsive core in X. citri were downregulated in bacteria isolated from transgenic leaves. DSF-dependent expression of engA was suppressed in cells exposed to xylem sap from transgenic plants. Thus, altered symptom development appears to be due to reduced expression of virulence genes because of the presence of antagonists of DSF signaling in X. citri in rpfF-expressing plants.

A comparative study of the prevalence of domestic violence in women requesting a termination of pregnancy and those attending the antenatal clinic.

OBJECTIVE: To compare the prevalence of domestic violence (DV) in women requesting antenatal care (ANC) and termination of pregnancy (TOP) in North East England. DESIGN: This was a cross-sectional comparative prevalence study using self-administered questionnaires, with women selected as opportunistic samples over a concurrent period. SETTING: The participants were screened anonymously and confidentially in the ANC and TOP clinics. SAMPLE: Pregnant women in the first trimester requesting a TOP or ANC. METHODS: The participants were screened for a history of DV using a modified version of the Abuse Assessment Screening tool. MAIN OUTCOME MEASURES: Prevalence of DV between ANC and TOP populations, and any differences in the characteristics of the women, such as age, level of education, or marital status. We aimed to determine the reasons for requesting a TOP. RESULTS: There were 507 respondents, with 233 attending ANC and 274 requesting a TOP. Of the ANC population, 219 completed the questionnaire. In the TOP population, all the questionnaires were fully or partially completed. Women requesting a TOP were six times as likely to suffer physical abuse in the current relationship (5.8 versus 0.9%; χ(2) = 10.2 (2); P < 0.05), and were five times as likely to suffer emotional abuse (9.9 versus 1.8%; χ(2) = 13.6 (2); P < 0.0001), than those attending ANC. Of the 274 women requesting a TOP, only ten mentioned DV as a contributing factor. CONCLUSIONS: There is a higher prevalence of DV in the TOP population than in the ANC population, but very few women stated that DV influenced their request for a TOP.

A comparison of maternal calcium and magnesium levels in pre-eclamptic and normotensive pregnancies: an observational case-control study.

OBJECTIVE: Supplementing pregnant women at high risk of developing pre-eclampsia with calcium may reduce the incidence of the disease. This study examines differences in serum and hair concentrations of calcium and magnesium between women with pre-eclamptic and normotensive pregnancies. DESIGN: Observational case-control study. SETTING: Two teaching hospitals in Cape Town, South Africa. POPULATION: Women with pre-eclamptic (N = 96) or normotensive (N = 96) pregnancies, who delivered a single, live infant. METHODS: Demographic and current pregnancy details were retrieved from clinical notes. Each participant completed a dietary questionnaire. Venous blood samples were taken from each participant to assess serum calcium and magnesium concentrations. Hair samples were obtained from all participants and calcium and magnesium levels were measured by inductively coupled plasma optical emission spectrometry (ICPOES). MAIN OUTCOME MEASURE: Hair and serum calcium and magnesium concentrations were compared between women with pre-eclamptic and normotensive pregnancies. RESULTS: Diet and socio-economic status in the two groups were similar. There was no significant difference in the hair calcium level between women with pre-eclamptic [1241 parts per million (ppm); range, 331-4654 ppm] and normotensive (1146 ppm; range, 480-4136 ppm) pregnancies (P = 0.5). Hair calcium levels in both groups were not affected by HIV infection. CONCLUSION: Woman with pre-eclampsia showed no difference in chronic calcium status relative to normotensive women. This finding does not support the current belief that the mechanism by which calcium supplementation reduces the risk of developing pre-eclampsia is by correcting a nutritional deficiency.

The use of fetal fibronectin in suspected pre-term labour.

Pre-term delivery is the leading cause of neonatal mortality and morbidity. The aim of this study is to determine the diagnostic accuracy of the fetal fibronectin (FF) test in predicting pre-term birth, the test interpretation and subsequent action taken in normal clinical practice in a busy tertiary centre setting. A total of 133 symptomatic women were included in the study and underwent the FF test. The use of tocolysis and corticosteroids were significantly greater in test-positive patients (p < 0.001). A negative test for detecting pre-term delivery within 10 days of the test was 100% sensitive with 100% negative predictive value (NPV). Our study has confirmed that a negative FF is an excellent short-term test to exclude pre-term delivery. The use of FF testing in routine clinical practice results in test characteristics similar to those found in research studies. This implies that the conclusions reached by researchers can be reliably translated into clinical practice.

Homelessness in pregnancy.

OBJECTIVES: To evaluate the association, if any, of homelessness or refuge accommodation on delivery and short term perinatal outcomes in an Irish tertiary maternity hospital. METHODS: A retrospective cohort study of 133 singleton pregnancies in women reporting to be homeless or living in refuge at their booking antenatal appointment between 2013 and 2022. Analysis compared sociodemographic characteristics and perinatal outcomes in this cohort to a reference population of 76,858 women with stable living arrangements. RESULTS: Women in the homeless/refuge population were statistically more likely to be single (75.2 % vs 39.5 %, p < 0.001), have an unplanned pregnancy (73.7 % vs 27.2 %, p < 0.001), report a history of psychiatric illness (42.9 % vs 22.4 %, p < 0.001), domestic violence (18.8 % vs 0.9 %, p < 0.001) alcohol consumption in pregnancy (3.0 % vs 0.8 %, p < 0.001) or smoking in pregnancy (41.3 % vs 9.7 %, p < 0.001). They were significantly more likely to have a preterm birth (adjusted OR 1.71 (1.01-2.87) p = 0.04). They also had a significantly lower median birth weight compared to the reference population (birthweight 3270 g vs 3420 g, p < 0.001). CONCLUSION: Women in the homeless and refuge population are more likely to experience poorer perinatal outcomes compared to women with stable living arrangements.

Cloning of the putative tumour suppressor gene for hereditary multiple exostoses (EXT1).

Hereditary multiple exostoses is an autosomal dominant disorder that is characterized by short stature and multiple, benign bone tumours. In a majority of families, the genetic defect (EXT1) is linked to the Langer-Giedion syndrome chromosomal region in 8q24.1. From this region we have cloned and characterized a cDNA which spans chromosomal breakpoints previously identified in two multiple exostoses patients. Furthermore, the gene harbours frameshift mutations in affected members of two EXT1 families. The cDNA has a coding region of 2,238 bp with no apparent homology to other known gene sequences and thus its function remains elusive. However, recent studies in sporadic and exostosis-derived chondrosarcomas suggest that the 8q24.1-encoded EXT1 gene may have tumour suppressor function.

Trends in obstetric research.

Never before have opportunities for research in reproductive sciences been so great and never before have relative resources been so limited. The challenge is to increase research studies as they form an integral part of good medicine. It has been suggested that the amount of obstetric medical research in the UK is declining, a major cause for concern, given the scale of morbidity caused by obstetrics disorders. The British Maternal and Fetal Medicine Society hold an annual congress with publication of all abstracts in the Journal of Obstetrics and Gynaecology. This study reviewed abstracts from 1997 to 2007 and categorised the research methodology, serving as a reasonable proxy for the volume of clinical investigation in UK obstetrics. Research studies requiring ethical approval dropped from 30% to 15% of the total in a decade. These figures prognosticate the future of research upon which many advances in medical care depend.

Obstetric consultant weekend on-call shift patterns have no effect on the management of spontaneous labour in a large maternity hospital.

Recent reports observe poorer healthcare outcomes during the weekend. Many attribute this weekend effect to a reduced consultant presence. This study evaluates differences in labour management on weekends vs weekdays. A total of 20,187 deliveries, all resulting from spontaneous labour, in the same large U.K. maternity hospital were examined. Labour management was analysed both for the department as a whole, and separately for each consultant, for differences in weekend and weekday practice. Results showed no statistically significant results of a difference in any of the measures analysed. On a weekend, deliveries were no more likely to be vaginal, p = 0.485, assisted, p = 0.771 or by caesarean section, p = 0.526. There was also no difference between individual consultants. It was concluded that for spontaneous labour, there is no difference in patient management on weekends vs weekdays, either in the department as a whole, or between individual consultants in our study.

Evaluation of a new Perinatal Mental Health Service in a University Maternity Hospital.

OBJECTIVES: Specialist Perinatal Mental Health Services (SPMHS) are a new development in Ireland. This service evaluation examined the impact of the introduction of a SPMHS multidisciplinary team (MDT) on prescribing practices and treatment pathways in an Irish maternity hospital. METHODS: Clinical charts were reviewed to collect data on all referrals, diagnoses, pharmacological and non-pharmacological interventions delivered in a SPMHS over a 3-week period in 2019. The findings were compared to the same 3-week period in 2020 following the expansion of the SPMHS MDT. RESULTS: In 2019 (n = 32) and 2020 (n = 47), most (75 and 79%, respectively) assessments were antenatal. The proportion of patients prescribed psychotropic medication within the SPMHS was not significantly different from 2019 (31%) to 2020 (23%), though more patients were already prescribed psychotropic medications at the time of referral (22% in 2019 v. 36% in 2020). There was an increase in MDT interventions in 2020 with more input from psychology, clinical nurse specialist (CNS), and social work intervention. Adherence to prescribing standards improved from 2019 to 2020. CONCLUSION: Prescribing patterns remained unchanged between 2019 and 2020. Improvement was observed in adherence to prescribing standards and there was increased provision of MDT interventions in 2020. Broader diagnostic categories were also used in 2020, possibly suggesting that the service is now providing more individualized care.

A cross-sectional survey of women's views and preferences for preterm birth screening and treatment.

OBJECTIVES: Primary outcomes were to determine; 1) the desire for more patient information from healthcare professionals on preterm birth (PTB) prevention 2) the desire for PTB screening surveillance or participation in research or 3) the acceptability of transvaginal ultrasound (TVUS) or vaginal examinations to predict spontaneous PTB. METHODS: A 19-question, piloted, self-administered survey was completed by unselected pregnant women in a tertiary maternity hospital in Dublin, Ireland. Data was collected to include maternal socio-demographics, past obstetric history, and current pregnancy details, in addition to views and preferences on PTB screening and preventative treatments. Statistical analysis to include binary and multinomial regression was performed by IBM SPSS Statistics for Windows (Version 29.0). RESULTS: 277 women completed the study survey. 9.4% of women had attended the preterm birth surveillance clinic (PSC). 75.1% of respondents indicated a preference for more information from healthcare professionals about PTB. 65% reported that TVUS and vaginal examinations were acceptable in pregnancy. The acceptability of antenatal examinations was significantly influenced by ethnicity; white European (OR 2.58, CI 1.12-5.95, p = 0.003) and Asian (OR 3.39, CI 1.18-9.67, p = 0.02). Discomfort (25.3%) and vaginal bleeding (11.9%) were the most frequently reported concerns about TVUS. 95.7% of unselected women indicated that they would accept treatment to prevent PTB. Vaginal progesterone (53.8%) was preferred treatment compared to cervical cerclage (15.9%) or cervical pessary (16.6%). 55.6% of respondents stated they attend or wish to attend for additional appointments or research opportunities for PTB screening. Women with a previous PTB or second trimester miscarriage were more likely to attend or wish to attend for PTB screening (OR 3.23, CI 1.34-7.79, p = 0.009). CONCLUSION: PTB is an important healthcare priority for pregnant women in Ireland. However, women require more information, counselling and reassurance about the utility and safety of TVUS in PSCs.

The duration of labor in spontaneous preterm deliveries: A retrospective observational study in a tertiary Irish maternity hospital.

BACKGROUND: Preterm deliveries account for 10% of all births, and are the most important cause of neonatal mortality globally. Despite their frequency, there is a paucity of information known about usual patterns of preterm labor, as previous studies which critically defined the normal progression of labor excluded preterm gestations. OBJECTIVE: To compare the durations of the first, second and third stages of spontaneous preterm labor in nulliparous and multiparous women at varying preterm gestations. METHODS: A retrospective observational study was undertaken of women admitted in spontaneous preterm labor from January 2017 to December 2020 with viable singleton gestations between 24 and 36 + 6 weeks' gestation who then proceeded to have a vaginal delivery. There were 512 cases following exclusion of preterm inductions of labor, instrumental vaginal deliveries, provider-initiated pre-labor Caesarean sections and emergency intrapartum Caesarean sections. The data was then examined to determine our outcomes of interest including the durations of the first, second and third stages of preterm labor, analyzing results by parity and gestation. For comparison, we reviewed data of term spontaneous labors and spontaneous vaginal deliveries during the same study period, identifying 8339 cases. FINDINGS: 97.6% of participants achieved a spontaneous cephalic vaginal delivery with the remainder undergoing an assisted breech birth. 5.7% of gestations delivered spontaneously between 24 + 0 and 27 + 6 weeks, with most births at gestations greater than 34 weeks (74%). The second stage duration (mean 15 vs 32 vs 32 mins respectively) was significantly different across the three gestation periods (p < 0.05), but was notably much quicker in extremely preterm labors. The first and third stage durations were similar between all gestational age groups with no statistically significant differences in results. There was a significant influence of parity on the first and second stages of labor, with multiparous women progressing more quickly than nulliparae (p < 0.001). CONCLUSION: The duration of spontaneous preterm labor is described. Multiparous women progress more quickly in the first and second stages of preterm labor than nulliparous women.

A retrospective observational study of labour ward work Intensity: The challenge of maternity staffing.

BACKGROUND: The UK and Ireland are facing significant challenges in the recruitment and retention of midwifery staff. Deficiencies in staffing, training and leadership have been cited as contributory factors to substandard care in both regional and global independent maternity safety reports. Locally, workforce planning is critical to maintaining 'one to one' care for all women in labor and to meet the peaks of daily birthing suite activity. OBJECTIVES: Analyze the variation in work intensity, defined by the mean number and range of births per midwifery working hours. METHODS: Retrospective observational study of birthing suite activity between 2017 and 2020. 30,550 singleton births were reported during the study period; however, 6529 elective Cesarean sections were excluded as these were performed during normal working hours by a separate operating theatre team. The times of 24,021 singleton births were organized into five proposed midwifery working rosters lasting eight or 12 h; A (00.00-07.59), B (08.00-15.59), C (16.00-23.59), D (20.00-0.759) and E (0.800-19.59). RESULTS: The number of births was comparable between the eight-hour and 12-hour work periods with a mean of five to six babies born per roster (range zero to 15). Work periods D and E lasting 12-hours both recorded a mean of eight births (range zero to 18). Hourly births ranged from a minimum of zero to a maximum of five births per hour (greater than seven times the mean), a number that was achieved 14 times during the study period. CONCLUSIONS: The mean number of births is consistent between normal working hours and unsociable 'on-call' periods, however there is an extreme range of activity within each midwifery roster. Prompt escalation plans remain essential for maternity services to manage unexpected increases in demand and complexity. WHAT IS ALREADY KNOWN ON THIS TOPIC: Shortfalls in staffing and inadequate workforce planning have been frequently cited in recent maternity safety reports as barriers to sustainable and safe maternity care. WHAT THIS STUDY ADDS: Our study shows that the mean number of births in a large tertiary center are consistent across day and night rosters. However, there are large fluctuations in activity during which births can exceed the number of available midwives. HOW THIS STUDY MIGHT AFFECT RESEARCH, PRACTICE OR POLICY: Our study reflects the sentiments of the Ockenden review and APPG report on safe maternity staffing. Investment in services and the workforce to aid recruitment and reduce attrition is essential to establish robust escalation plans, including the deployment of additional staff in the event of extreme service pressures.

Cell free DNA screening for fetal aneuploidy in Ireland: An observational study of outcomes following insufficient fetal fraction.

OBJECTIVES: To determine maternal factors associated with low fetal fraction (FF). To determine the proportion of women who receive a result from repeat non-invasive prenatal testing (NIPT) testing. To identify any significant associations between pregnancy interventions or outcomes and low FF. STUDY DESIGN: Retrospective observational study of 4465 women undergoing antenatal screening by targeted cell free DNA (cfDNA) testing at an Irish tertiary maternity hospital between January 2017 and December 2022. Patients who failed to obtain a result after the first NIPT were analyzed in two cohorts; those who received a result on a repeat sample and those who failed to ever achieve a result despite a second, third or fourth cfDNA test. RESULTS: Risk of insufficient FF significantly increased with elevated maternal BMI (OR 1.07; 95% CI 1.01-1.13, p = 0.03) and in-vitro fertilization (IVF) (OR 3.4; 95% CI 1.19-9.4, p = 0.02). Women with no result were more likely to have diagnostic invasive testing (p < 0.01), but had no increased risk of aneuploidy. Repeated failed NIPT attempts due to low FF were significantly associated with the subsequent development of hypertensive diseases of pregnancy (p = 0.03). Greater than 70% of patients who were unsuccessful in a first or second attempt at NIPT due to low FF yielded a result following a second or third sample. CONCLUSIONS: High BMI and IVF conceptions are greater contributors to low FF than fetal aneuploidy. Repeating NIPT yields a result in greater than 70% of cases. WHAT'S ALREADY KNOWN ABOUT THIS TOPIC?: Fetal fraction (FF) in prenatal cfDNA testing is influenced by maternal and pregnancy factors including body mass index (BMI) and IVF. Low FF has been associated with adverse pregnancy outcomes including fetal aneuploidy and hypertensive diseases of pregnancy. WHAT DOES THIS STUDY ADD?: In a large Irish population, increasing maternal BMI and in-vitro fertilization are the most significant contributors to repeated test failures due to low FF. Greater than 70% of patients with test failure due to low FF will receive a result on 2nd and 3rd NIPT attempts. Patients with no result from NIPT were more likely to undergo diagnostic invasive testing but the risk of aneuploidy was not significantly increased.

An analysis of the performance of UK medical graduates in the MRCOG Part 1 and Part 2 written examinations.

BACKGROUND: There is a lack of evidence on whether graduates from different medical schools perform differently in postgraduate examinations. OBJECTIVE: To evaluate the variations in performance of UK medical graduates in Member of the Royal College of Obstetricians and Gynaecologists (MRCOG) examination. METHODS: A retrospective analysis of performance of 1335 doctors graduating in UK medical schools who entered the Part 1 MRCOG and 822 doctors taking the Part 2 MRCOG written examination for the first time between 1998 and 2008. The main outcome measures were to evaluate medical school effects, gender effects and academic performance effect. RESULTS: Graduates of UK medical schools performed differently in the Part 1 and Part 2 written MRCOG examination. The graduates of Oxford (pass rate 82.6%), Cambridge (75%), Bristol (59.3%) and Edinburgh (57.5%) performed significantly better and the graduates of Liverpool (26.8%), Southampton (21.8%) and Wales (18.2%) performed significantly worse than the remaining cohort in the Part 1 examination. The candidates of Newcastle (88.9%), Oxford (82.4%), Cambridge (81%) and Edinburgh (78.2%) performed significantly better and the graduates of Glasgow (49.2%) and Leicester (36.2%) have significantly underperformed compared with the remaining cohort in Part 2 written examination. There was no difference in the success rates of male (47.5%) and female (42.0%) candidates in the Part 1; however, female candidates had a significantly better success rate in the Part 2 written examination than male candidates (65.6% vs 52.9%). CONCLUSION: These results show that there is variation in performance among the graduates from different medical schools in the Part 1 and Part 2 MRCOG written examination.

Time of birth and delivery outcomes: a retrospective cohort study.

Studies performed previously have attributed increased risk of intrapartum anoxia to delivery outside the normal working week. We sought to determine if the time of delivery effects outcome with respect to delivery characteristics and neonatal condition at birth. We identified a cohort of 14,426 deliveries over a 3-year period. These were analysed as three groups: night (00:00-07:59); day (08:00-15:59) and twilight (16:00-23:59). We found significantly more deliveries at night 36.3% (n = 5,240; p =0.01). At night, there were non-significantly more 'normal' deliveries compared with during the day and twilight hours (78.4%, 76.2%, 77.3%, respectively; p = 0.27). The caesarean section rate did not differ significantly between the groups (14.2%, 15.6%, 14.8%; p = 0.147). There were significantly fewer assisted deliveries in the night-time period (6.6%, 8.0%, 7.3%; p = 0.03). Women who delivered at night were more likely to have a normal delivery and less likely to have obstetric intervention in the form of assisted or operative delivery. There were no significant differences in the need for resuscitation to be performed at birth across the three groups (14.3%, 13.3%, 14.8%; p = 0.111). There was no significant difference in the numbers of babies directly admitted to the neonatal intensive care unit (NICU) (5.9%, 6.8%, 6.3%; p = 0.198) in the three respective time periods.

Methadone use in pregnancy: evidence of progression in the severity of addiction.

The number of opiate users is well documented; however, the severity of opiate use has received little attention. This retrospective study in a North of England hospital updates the progression in the severity of addiction in pregnancy. Patients treated were reviewed and the doses of prescribed methadone documented. Historical data were also used for comparison. The severity in addiction in pregnancy was assessed by the woman's drug usage expressed as the daily dose of prescribed methadone at the end of pregnancy. From 2001 to 2008 there was an increase in the mean dose of methadone prescribed at delivery from 28.2 ml/day in 2001 to 57.9 ml/day in 2008. Historically, the use was 27.3 ml/day in 1992-1996 and 32.4 ml/day in 1997-2003. No trend was noted in the number of pregnant users. In conclusion, we observed no recent increase in the number of methadone users presenting, but the severity of drug usage in pregnancy has increased.