A longitudinal study of the experiences and psychological well-being of Indian surrogates

Study question: What is the psychological well-being of Indian surrogates during and after the surrogacy pregnancy? Summary answer: Surrogates were similar to a matched group of expectant mothers on anxiety and stress. However, they scored higher on depression during and after pregnancy. What is known already: The recent ban on trans-national commercial surrogacy in India has led to urgent policy discussions regarding surrogacy. Whilst previous studies have reported the motivations and experiences of Indian surrogates no studies have systematically examined the psychological well-being of Indian surrogates, especially from a longitudinal perspective. Previous research has shown that Indian surrogates are motivated by financial payment and may face criticism from their family and community due to negative social stigma attached to surrogacy. Indian surrogates often recruited by agencies and mainly live together in a “surrogacy house.” Study design, size, duration: A longitudinal study was conducted comparing surrogates to a matched group of expectant mothers over two time points: (a) during pregnancy (Phase1: 50 surrogates, 70 expectant mothers) and (b) 4–6 months after delivery (Phase 2: 45 surrogates, 49 expectant mothers). The Surrogates were recruited from a fertility clinic in Mumbai and the matched comparison group was recruited from four public hospitals in Mumbai and Delhi. Data collection was completed over 2 years. Participants/materials, setting, methods: Surrogates and expectant mothers were aged between 23 and 36 years. All participants were from a low socio-economic background and had left school before 12–13 years of age. In-depth faceto-face semi-structured interviews and a psychological questionnaire assessing anxiety, stress and depression were administered in Hindi to both groups. Interviews took place in a private setting. Audio recordings of surrogate interviews were later translated and transcribed into English. Main results and the role of chance: Stress and anxiety levels did not significantly differ between the two groups for both phases of the study. For depression, surrogates were found to be significantly more depressed than expectant mothers at phase 1 (p = 0.012) and phase 2 (p = 0.017). Within the surrogacy group, stress and depression did not change during and after pregnancy. However, a non-significant trend was found showing that anxiety decreased after delivery (p = 0.086). No participants reported being coerced into surrogacy, however nearly all kept it a secret from their wider family and community and hence did not face criticism. Surrogates lived at the surrogate house for different durations. During pregnancy, 66% (N = 33/50) reported their experiences of the surrogate house as positive, 24% (N = 12/50) as negative and 10% (N = 5/50) as neutral. After delivery, most surrogates (66%, N = 30/45) reported their experiences of surrogacy to be positive, with the remainder viewing it as neutral (28%) or negative (4%). In addition, most (66%, N = 30/45) reported that they had felt “socially supported and loved” during the surrogacy arrangement by friends in the surrogate hostel, clinic staff or family. Most surrogates did not meet the intending parents (49%, N = 22/45) or the resultant child (75%, N = 34/45). Limitations, reasons for caution: Since the surrogates were recruited from only one clinic, the findings may not be representative of all Indian surrogates. Some were lost to follow-up which may have produced sampling bias. Wider implications of the findings: This is the first study to examine the psychological well-being of surrogates in India. This research is of relevance to current policy discussions in India regarding legislation on surrogacy. Moreover, the findings are of relevance to clinicians, counselors and other professionals involved in surrogacy. Trial registration number: N/A

Harnessing endogenous human ADARs for RNA repair

Site‐directed adenosine-to-inosine (A-to-I) RNA editing is a novel transcriptome engineering approach that allows to recode genetic information at RNA level by specific deamination of adenosines. Because the resulting inosine is read as guanosine by cellular machines and during translation, site-directed RNA editing (SDRE) does ultimately introduce A-to-G base substitutions into mRNAs. This opens up the opportunity to recode Start- and Stop-codons, splice signals, miRNA recognition sites, and 12 of the 20 canonical amino acids. Thus SDRE allows a wide range of interventions, including the manipulation of residues relevant for signaling or protein function, and the ability to correct G-to-A point mutations. Our R/G guideRNA SDRE system allows to steer wild-type human ADAR (adenosine deaminase acting on RNA) enzymes towards selected target adenosines within user-defined mRNAs. The genetically encodable R/G gRNA design, which was developed during this thesis based on successive rounds of rational design, allows for the first time to recruit endogenous human ADAR enzymes. In contrast to all other SDRE approaches, the R/G gRNA system does therefore not require the overexpression of an editase. Transcriptome engineering can now be achieved by the simple application of a plasmid or adenovirus encoded short guideRNA. During this thesis, several fundamental questions, which had to precede the further development of SDRE into a therapeutic application, could be solved. The R/G gRNA system became the first SDRE approach to prove editing of endogenous mRNAs. In addition, it was used to recode the recessive PINK1 W437X amber loss-of-function mutation in HeLa cells and could show that SDRE allows the functional rescue of PINK1-Parkin-mediated mitophagy, which is linked to the etiology of Parkinson's disease. Our R/G gRNAs have high potential to become a next generation drug for the precise correction of disease-causing point mutations and the tuneable manipulation of protein function. In doing so, they would complement existing genome and transcriptome manipulation strategies by enabling interventions out of reach for currently available molecular tools.Gerichtete A-nach-I RNA-Editierung ist ein neues Transkriptom-Manipulationsverfahren, das die Desaminierung ausgewählter Adenosine und damit deren Umwandlung zu Inosin ermöglicht. Da Inosin während der Transkription und Translation als Guanosin erkannt wird, können mit dieser Methode genetische Informationen auf der RNA-Ebene bearbeitet werden. Im Grunde fügt das Verfahren also ge-zielt A-nach-G Punktmutationen in mRNAs ein und erlaubt damit die Umcodierung von Start- und Stopp-Codons, Spleißstellen, miRNA-Bindestellen, sowie 12 der 20 kanonischen Aminosäuren. Hierdurch wird eine Vielzahl an Eingriffen möglich, wie die Korrektur von G-nach-A Punktmutationen, die Manipulation von Proteinfunktionen durch Änderung relevanter Aminosäuren, oder die Modulation zellulärer Signal-kaskaden. Das von uns entwickelte R/G guideRNA System dirigiert humanes Wildtyp-ADAR (adenosine deaminase acting on RNA) zu bestimmten Ziel-Adenosinen in ausgewählten mRNAs. Durch sukzessives rationales Design wurde im Zuge dieser Arbeit, ein genetisch codierbares R/G gRNA Design entwickelt, das es erstmals erlaubt endogenes humanes ADAR zu rekrutieren. Im Gegensatz zu allen anderen gegenwärtig verfügbaren gerichteten RNA-Editierungsverfahren, ist es daher nicht auf die Überexpression einer Editase angewiesen. Hierdurch kann nun genetische Information auf Ebene des Transkriptoms durch simples Verabreichen kurzer Plasmid- oder Adenovirus-codierter guideRNAs manipuliert werden. Zusätzlich konnten in dieser Arbeit grundlegende Fragen beantwortet werden, die der Weiterentwicklung von gerichteter RNA-Editierung zu einer therapeutischen Anwendung vorangehen mussten. Es konnte erstmals gezeigt werden, dass durch das R/G gRNA-System auch die Editierung endogener mRNAs möglich ist. Des Weiteren wurde die rezessive PINK1-W437X-Amber Funktionsverlustmutation in HeLa Zellen korrigiert. Das Verfahren erlaubte es hierbei, die Funktion des durch die genannte Mutation gestörten PINK1-Parkin-vermittelten Mitophagie-Signalwegs, welcher mit der Ätiologie der Parkinsonkrankheit verknüpft ist, wiederherzustellen. Unsere R/G gRNAs haben das Potential als völlig neue Generation von Medikamenten beispielsweise für die Behandlung von Punktmutationen oder therapeutische Regulierung von Proteinfunktionen eingesetzt zu werden. Hierbei würden bestehende Ansätze zur Genom- und Transkriptom-Manipulation durch bisher undurchführbare Eingriffe auf RNA-Ebene ergänzt

Smoking and Second Hand Smoking in Adolescents with Chronic Kidney Disease: A Report from the Chronic Kidney Disease in Children (CKiD) Cohort Study

The goal of this study was to determine the prevalence of smoking and second hand smoking [SHS] in adolescents with CKD and their relationship to baseline parameters at enrollment in the CKiD, observational cohort study of 600 children (aged 1-16 yrs) with Schwartz estimated GFR of 30-90 ml/min/1.73m2. 239 adolescents had self-report survey data on smoking and SHS exposure: 21 [9%] subjects had “ever” smoked a cigarette. Among them, 4 were current and 17 were former smokers. Hypertension was more prevalent in those that had “ever” smoked a cigarette (42%) compared to non-smokers (9%), p\u3c0.01. Among 218 non-smokers, 130 (59%) were male, 142 (65%) were Caucasian; 60 (28%) reported SHS exposure compared to 158 (72%) with no exposure. Non-smoker adolescents with SHS exposure were compared to those without SHS exposure. There was no racial, age, or gender differences between both groups. Baseline creatinine, diastolic hypertension, C reactive protein, lipid profile, GFR and hemoglobin were not statistically different. Significantly higher protein to creatinine ratio (0.90 vs. 0.53, p\u3c0.01) was observed in those exposed to SHS compared to those not exposed. Exposed adolescents were heavier than non-exposed adolescents (85th percentile vs. 55th percentile for BMI, p\u3c 0.01). Uncontrolled casual systolic hypertension was twice as prevalent among those exposed to SHS (16%) compared to those not exposed to SHS (7%), though the difference was not statistically significant (p= 0.07). Adjusted multivariate regression analysis [OR (95% CI)] showed that increased protein to creatinine ratio [1.34 (1.03, 1.75)] and higher BMI [1.14 (1.02, 1.29)] were independently associated with exposure to SHS among non-smoker adolescents. These results reveal that among adolescents with CKD, cigarette use is low and SHS is highly prevalent. The association of smoking with hypertension and SHS with increased proteinuria suggests a possible role of these factors in CKD progression and cardiovascular outcomes