Absence of maternal methylation in biparental hydatidiform moles from women with NLRP7 maternal-effect mutations reveals widespread placenta-specific imprinting

Familial recurrent hydatidiform mole (RHM) is a maternal-effect autosomal recessive disorder usually associated with mutations of the NLRP7 gene. It is characterized by HM with excessive trophoblastic proliferation, which mimics the appearance of androgenetic molar conceptuses despite their diploid biparental constitution. It has been proposed that the phenotypes of both types of mole are associated with aberrant genomic imprinting. However no systematic analyses for imprinting defects have been reported. Here, we present the genome-wide methylation profiles of both spontaneous androgenetic and biparental NLRP7 defective molar tissues. We observe total paternalization of all ubiquitous and placenta-specific differentially methylated regions (DMRs) in four androgenetic moles; namely gain of methylation at paternally methylated loci and absence of methylation at maternally methylated regions. The methylation defects observed in five RHM biopsies from NLRP7 defective patients are restricted to lack-of-methylation at maternal DMRs. Surprisingly RHMs from two sisters with the same missense mutations, as well as consecutive RHMs from one affected female show subtle allelic methylation differences, suggesting inter-RHM variation. These epigenotypes are consistent with NLRP7 being a maternal-effect gene and involved in imprint acquisition in the oocyte. In addition, bioinformatic screening of the resulting methylation datasets identified over sixty loci with methylation profiles consistent with imprinting in the placenta, of which we confirm 22 as novel maternally methylated loci. These observations strongly suggest that the molar phenotypes are due to defective placenta-specific imprinting and over-expression of paternally expressed transcripts, highlighting that maternal-effect mutations of NLRP7 are associated with the most severe form of multi-locus imprinting defects in humans

Stimulatory Effects of Arsenic-Tolerant Soil Fungi on Plant Growth Promotion and Soil Properties

Fifteen fungi were obtained from arsenic-contaminated agricultural fields in West Bengal, India and examined for their arsenic tolerance and removal ability in our previous study. Of these, the four best arsenic-remediating isolates were tested for plant growth promotion effects on rice and pea in the present study. A greenhouse-based pot experiment was conducted using soil inocula of individual fungi. The results indicated a significant (P<0.05) increase in plant growth and improvement of soil properties in inoculated soils compared to the control. A significant increase in plant growth was recorded in treated soils and varied from 16–293%. Soil chemical and enzymatic properties varied from 20–222 % and 34–760%, respectively, in inoculated soil. Plants inoculated with inocula of Westerdykella and Trichoderma showed better stimulatory effects on plant growth and soil nutrient availability than Rhizopus and Lasiodiplodia. These fungi improved soil nutrient content and enhanced plant growth. These fungi may be used as bioinoculants for plant growth promotion and improved soil properties in arsenic-contaminated agricultural soils. Key words: pea, plant growth-promoting fungi, rice, soil enzymes, arsenic-contaminated soils Chemical fertilizers, agrochemicals and pesticides have been intensively applied to soils to increase crop production to meet food demands all over the world; however, these are disruptive to soil biological processes. Exploring nove

COVID-19 vaccine hesitancy:a midwifery survey into attitudes towards the COVID-19 vaccine

BACKGROUND: Ethnically minoritised people have been disproportionately affected by the COVID-19 pandemic. Emerging evidence suggests a lower uptake of the vaccine in ethnically minoritised people, particularly Black females of reproductive age. Unvaccinated pregnant women are high risk for morbidity and mortality from COVID-19. Midwives are the principal healthcare professionals responsible for counselling the pregnant population on decisions relating to vaccine uptake. The aim of this study was to explore midwifery uptake of and attitudes towards the COVID-19 vaccine in two ethnically diverse areas. METHODS: A 45-point questionnaire was circulated over a six-week period to midwives employed in two teaching hospitals in England; London (Barts Health NHS Trust) and Sussex (Brighton and Sussex University Hospitals NHS Trust (BSUH)). A total of 378 out of 868 midwives responded. Results were analysed to determine vaccine uptake as well as factors influencing vaccine hesitancy and decision-making between the two trusts and ethnic groups. Thematic analysis was also undertaken. RESULTS: Midwives of Black ethnicities were over 4-times less likely to have received a COVID-19 vaccine compared to midwives of White ethnicities (52% vs 85%, adjusted OR = 0.22, p = < 0.001). Overall, there were no significant differences between trusts in receipt of the COVID-19 vaccine (p = 0.13). Midwives at Barts Health were significantly more likely to have tested positive for COVID-19 compared to midwives at BSUH (adjusted OR = 2.55, p = 0.007). There was no statistical difference between ethnicities in testing positive for COVID-19 (p = 0.86). The most common concerns amongst all participants were regarding the long-term effect of the vaccine (35%), that it was developed too fast (24%), having an allergic reaction (22%) and concerns about fertility (15%). Amongst unvaccinated midwives, those of Black ethnicity had a higher occurrence of concern that the vaccine contained meat / porcine products (adjusted OR = 5.93, p = 0.04) and that the vaccine would have an adverse effect on ethnic minorities (adjusted OR = 4.42, p = 0.03). CONCLUSION: This study highlights the significantly higher level of vaccine hesitancy amongst Black midwives and offer insights into midwives’ concerns. This can facilitate future targeted public health interventions. It is essential that vaccine hesitancy amongst midwifery staff is addressed to improve vaccine uptake in the pregnant population

Absence of Maternal Methylation in Biparental Hydatidiform Moles from Women with NLRP7 Maternal-Effect Mutations Reveals Widespread Placenta-Specific Imprinting

Familial recurrent hydatidiform mole (RHM) is a maternal-effect autosomal recessive disorder usually associated with mutations of the NLRP7 gene. It is characterized by HM with excessive trophoblastic proliferation, which mimics the appearance of androgenetic molar conceptuses despite their diploid biparental constitution. It has been proposed that the phenotypes of both types of mole are associated with aberrant genomic imprinting. However no systematic analyses for imprinting defects have been reported. Here, we present the genome-wide methylation profiles of both spontaneous androgenetic and biparental NLRP7 defective molar tissues. We observe total paternalization of all ubiquitous and placentaspecific differentially methylated regions (DMRs) in four androgenetic moles; namely gain of methylation at paternally methylated loci and absence of methylation at maternally methylated regions. The methylation defects observed in five RHM biopsies from NLRP7 defective patients are restricted to lack-of-methylation at maternal DMRs. Surprisingly RHMs from two sisters with the same missense mutations, as well as consecutive RHMs from one affected female show subtle allelic methylation differences, suggesting inter-RHM variation. These epigenotypes are consistent with NLRP7 being a maternal-effect gene and involved in imprint acquisition in the oocyte. In addition, bioinformatic screening of the resulting methylation datasets identified over sixty loci with methylation profiles consistent with imprinting in the placenta, of which we confirm 22 as novel maternally methylated loci. These observations strongly suggest that the molar phenotypes are due to defective placenta-specific imprinting and over-expression of paternally expressed transcripts, highlighting that maternal-effect mutations of NLRP7 are associated with the most severe form of multi-locus imprinting defects in humans

Identification of additional placenta-specific imprinted DMRs in RHM samples.

<p>(A) A heatmap for the β_mean of the Infinium probes with a methylation difference (>20%, minimum 3 consecutive probes) in RHMs associated with maternal effect <i>NLRP7</i> mutations compared to control placental biopsies. (B) Schematic representation of the methylation-sensitive <i>Hpa</i>II genotyping assay. (C) Methylation profiles as determined by methylation-sensitive genotyping and (D) bisulfite PCR and subcloning on placenta and somatic tissue DNA samples at the <i>SCIN</i>, <i>ST8AIA1</i> and <i>CABIN1</i> promoters. Note that the samples used for methylation-sensitive genotyping and bisulphite PCR maybe different to highlight that methylation is not associated with genotype but parental origin.</p

Description of <i>NLRP7</i> mutations with methylation and expression profiling of imprinted loci.

<p>(A) Confirmation of recessive <i>NLRP7</i> mutations in female patients and heterozygous status in the RHM samples. The asterisk (*) on the electropherogram highlights the position of the mutation. For patient 3 the position of the deletion is shown. (B) Circular heat map of the 616 Infinium array probes mapping to 36 ubiquitously imprinted DMRs. The inner circle represents the methylation values of androgenetic HMs, the middle circles normal placental biopsies and the outer circle the RHMs associated with maternal-effect <i>NLRP7</i> mutations. (C) Confirmation of the methylation profile of the <i>NLRP7</i> mutated RHMs at the <i>NAP1L5</i>, <i>PEG10</i>, <i>RB1</i>, <i>L3MBTL1</i> and <i>H19</i> DMRs by bisulphite PCR and subcloning. Each circle represents a single CpG dinucleotide on a DNA strand, a methylated cytosine (●) or an unmethylated cytosine (○). For clarity, only the first 10 CpG dinucleotides from each amplicon are shown with the letters in the parentheses indicating SNP genotype. (D) Allelic expression analysis of imprinted genes <i>NAP1L5</i>, <i>HYMAI</i>, <i>PEG10</i> and <i>PEG3</i> in control placenta samples (PL) and <i>NLRP7</i>-mutated moles (RHM).</p