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

Assessment of DNA structure and integrity in the human spermatozoon

Male fertility is routinely assessed by basic semen analysis, but this is poorly predictive of fertility outcome. One promising advanced diagnostic is assessing sperm DNA damage, but there is a lack of method standardisation, clinical thresholds and comparison data for different assays. The objective of this study was to validate standardised methods for the assessment of sperm DNA quality using TUNEL, Acridine orange (AO) and Chromomycin A3 (CMA3) in slide-based assays; and assess prognostic value for fertility and miscarriage. The data obtained reveals that using differing optimal mounting solution for different assays is key to reliable results. The use of DNA fragmentation inductors such as DNase and hydrogen peroxide in donor samples confirmed and validated the use of AO and TUNEL for detecting DNA damaged cells. A novel semi-automated computer-based scoring system for fluorescence microscopy images was devised and compared with visual operator results for intra-assay variability of AO and TUNEL assays. This system allowed objective and consistent results free of operator subjectivity. The assessment of TUNEL, AO and CMA3 values in a subset of patients from the HABSelect trial showed no correlation between the assays corroborating that different assays measure different aspects of DNA quality. The number of patient samples assessed were insufficiently powered to draw firm conclusions related to clinical outcome, but we believe they are useful in making a case for further investigations in the field

Data mining applied to the Varicocele condition

O sistema de saúde guarda cada vez mais informação dos seus utentes o que dificulta ou até impossibilita a descoberta de novos conhecimentos só com as técnicas usualmente utilizadas, i.e., as tradicionais técnicas estatísticas. De facto, os investigadores clínicos têm sentido uma crescente necessidade em extrair novos conhecimentos para continuadamente contribuir para o melhoramento dos serviços de saúde prestados. Essa necessidade tem vindo a ser colmatada com a aplicação de um processo, chamado “data mining”, que auxilia, através da aplicação de diversas técnicas (i.e., classificação, clustering, associação, etc.), a descoberta de padrões de dados vistos como interessantes, mas ocultados com as tradicionais técnicas estatísticas. A área da infertilidade masculina já começou a aplicar o data mining, por exemplo, através da aplicação da técnica de classificação para prever o sucesso de uma técnica de Procriação Medicamente Assistida. Contudo, o varicocelo - um síndrome anatómico de varizes escrotais caracterizado pela dilatação das veias que drenam o sangue da região dos testículos que em certos casos dá origem à infertilidade - não foi ainda explorado com uma técnica de data mining. A sua prevalência atinge 40% dos homens tratados por infertilidade, sendo que a infertilidade masculina abrange 50% das causas da infertilidade de um casal. A correção do varicocelo pode ser alcançada com um tratamento radiológico chamado embolização, que tem por objetivo desvitalizar as veias dilatadas através da introdução de substâncias terapêuticas na circulação sanguínea. Neste contexto, este trabalho teve os seguintes principais objetivos: i) averiguar o sucesso da correção do varicocelo com a técnica da embolização através da identificação de algum melhoramento na média dos valores dos parâmetros seminais ou das categorias seminais com recurso a técnicas estatísticas inferenciais (i.e. ANOVA e Chi-quadrado); ii) predizer o sucesso da embolização com técnicas de classificação através da aplicação do decision tree do RapidMiner e do algoritmo W-J48; iii) identificar padrões que caracterizam os pacientes embolizados com a técnica de clustering através do algoritmo K-Means e eleger as relações de atributos que ocorrem mais frequentemente através da técnica de associação com o algoritmo FP-Growth. Este processo de análise de dados seguiu a metodologia Cross-Industry Standard Process for Data mining (CRISP-DM) aplicando-a à análise de uma amostra de 293 homens inférteis descritos com 64 atributos que foram submetidos à embolização no Centro Hospitalar e Universitário de Coimbra (CHUC) entre Janeiro de 2007 e Abril de 2016. Os resultados obtidos indicam que a embolização melhora significativamente a média das concentrações de espermatozoides até 12 meses e de suas morfologias até 6 meses depois da embolização (ANOVA p<0.05) o que permite fundamentar o interesse em prever o sucesso desta técnica terapêutica. Sua previsão computarizada com a árvore de decisão do RapidMiner permitiu prever com uma Accuracy e F-measure de 70.59% e uma AUC de 0.750 que a probabilidade condicional de engravidar tendo um homem com uma severidade baixa ou média do varicocelo e uma parceira entre os 24 e 33 anos inclusive é de 70.83%. Também se viu que a frequência relativa, de pacientes com uma concentração de espermatozoides normal 3 meses depois da embolização e uma motilidade progressiva normal destes antes do tratamento, é mais alta em grupos de pacientes que raramente trabalham em ambientes tóxicos. Estes resultados permitem contribuir para as investigações em curso no domínio da infertilidade, assim como nidentificação de medidas que permitem um maior auxílio na descoberta do conhecimento. Nomeadamente, vimos que a aplicação conjunta dos algoritmos de data mining com as técnicas estatísticas inferenciais, assim como a aplicação de diversas técnicas de data mining (i.e., classificação, clustering e associação), potencia a descoberta do conhecimento em dados clínicos

Studies on the modification of ram spermatozoa by ejaculation and cryopreservation and the effects of Binder of Sperm Proteins

Frozen thawed ram spermatozoa must be deposited directly into the uterus via laparoscopy as they are unable to traverse the cervix in sufficient numbers to achieve acceptable fertility. Seminal plasma has been heralded as a possible solution to this problem, as it appears to enhance sperm cervical transit. However, the mechanism by which seminal plasma, or indeed cryopreservation, alters spermatozoa to subsequently influence cervical transit is unknown. This thesis explores how seminal plasma and cryopreservation affect the structure and function of ram spermatozoa, with the ultimate aim of improving the in vivo performance of cryopreserved ram semen. The findings of this thesis demonstrate that the physiological process of ejaculation and the artificial practice of cryopreservation significantly alter the molecular profile and function of spermatozoa. Seminal plasma exposure confers additional proteins, alters the sperm glycocalyx, protects spermatozoa from immune cells and regulates the timing of capacitation. Cryopreservation also alters the sperm proteome and glycocalyx, possibly increasing susceptibility of this sperm type to dynamic environmental pressures within the female reproductive tract. Further investigation of the degree to which BSPs benefit ram spermatozoa in the female tract is warranted given their significant effects on sperm cryosurvival and physiology demonstrated herein. The importance of this protein family in ram sperm physiology offers a potential avenue for future improvements in the fertility of frozen thawed ram spermatozoa following cervical insemination

Molecular and physical interactions of human sperm with female tract secretions

To achieve fertilisation, human sperm have to navigate and interact with the female reproductive tract (FRT) on molecular and mechanical levels. The current knowledge of some aspects of both types of interactions are limited and they were examined in this research. Proteomic analysis of crude and depleted human follicular fluid (hFF) by three proteomic approaches identified 479 hFF-proteins of which 22% were novel. A table of hFF-proteins, compiled from twenty-four hFF proteomic studies, resulted in 1586 hFF proteins; a resource for folliculogenesis and discovery of hFF biomarkers. A comparative proteomic study of media-capacitated human sperm versus capacitated sperm in the presence of hFF revealed certain hFF proteins were acquired by sperm during capacitation. Comparative metabolomics revealed some elevated metabolites in the sperm capacitation-media following 6-hour incubation compared to 1-hour swim-up, which may have relevance to sperm energy metabolism and potentially to sperm signalling mechanisms in the FRT causing remodelling for fertilisation and preparation for implantation. Sperm micro-particle image-velocimetry revealed an average fluid velocity around the motile sperm of ≤ 25- 45 μm/s and ≤ 20-35 μm/s in low and high viscosity media respectively. The averaged fluid vorticity manifested a trail of spatially confined mixing of the fluid surroundings motile sperm. To the best of our knowledge, this work is the first to conduct a comparative proteomic analysis of human sperm versus sperm-hFF interaction and to perform metabolomics analysis of human sperm capacitation media at two time points. The novel perspective of mechanical aspects of sperm motility by studying fluid velocity and vorticity around motile sperm adds a new approach to the study of sperm motility and chemotaxis. In conclusion, sperm-FRT interactions involve complex molecular and physical interactions and regulatory events. Further research of these interactions may enhance our understanding of potential applications for improved assisted reproductive techniques’ outcome and possible diagnostic approaches of infertility

Meiosis

Meiosis, the process of forming gametes in preparation for sexual reproduction, has long been a focus of intense study. Meiosis has been studied at the cytological, genetic, molecular and cellular levels. Studies in model systems have revealed common underlying mechanisms while in parallel, studies in diverse organisms have revealed the incredible variation in meiotic mechanisms. This book brings together many of the diverse strands of investigation into this fascinating and challenging field of biology

Psr1p interacts with SUN/sad1p and EB1/mal3p to establish the bipolar spindle

Regular Abstracts - Sunday Poster Presentations: no. 382During mitosis, interpolar microtubules from two spindle pole bodies (SPBs) interdigitate to create an antiparallel microtubule array for accommodating numerous regulatory proteins. Among these proteins, the kinesin-5 cut7p/Eg5 is the key player responsible for sliding apart antiparallel microtubules and thus helps in establishing the bipolar spindle. At the onset of mitosis, two SPBs are adjacent to one another with most microtubules running nearly parallel toward the nuclear envelope, creating an unfavorable microtubule configuration for the kinesin-5 kinesins. Therefore, how the cell organizes the antiparallel microtubule array in the first place at mitotic onset remains enigmatic. Here, we show that a novel protein psrp1p localizes to the SPB and plays a key role in organizing the antiparallel microtubule array. The absence of psr1+ leads to a transient monopolar spindle and massive chromosome loss. Further functional characterization demonstrates that psr1p is recruited to the SPB through interaction with the conserved SUN protein sad1p and that psr1p physically interacts with the conserved microtubule plus tip protein mal3p/EB1. These results suggest a model that psr1p serves as a linking protein between sad1p/SUN and mal3p/EB1 to allow microtubule plus ends to be coupled to the SPBs for organization of an antiparallel microtubule array. Thus, we conclude that psr1p is involved in organizing the antiparallel microtubule array in the first place at mitosis onset by interaction with SUN/sad1p and EB1/mal3p, thereby establishing the bipolar spindle.postprin

Removal of antagonistic spindle forces can rescue metaphase spindle length and reduce chromosome segregation defects

Regular Abstracts - Tuesday Poster Presentations: no. 1925Metaphase describes a phase of mitosis where chromosomes are attached and oriented on the bipolar spindle for subsequent segregation at anaphase. In diverse cell types, the metaphase spindle is maintained at a relatively constant length. Metaphase spindle length is proposed to be regulated by a balance of pushing and pulling forces generated by distinct sets of spindle microtubules and their interactions with motors and microtubule-associated proteins (MAPs). Spindle length appears important for chromosome segregation fidelity, as cells with shorter or longer than normal metaphase spindles, generated through deletion or inhibition of individual mitotic motors or MAPs, showed chromosome segregation defects. To test the force balance model of spindle length control and its effect on chromosome segregation, we applied fast microfluidic temperature-control with live-cell imaging to monitor the effect of switching off different combinations of antagonistic forces in the fission yeast metaphase spindle. We show that spindle midzone proteins kinesin-5 cut7p and microtubule bundler ase1p contribute to outward pushing forces, and spindle kinetochore proteins kinesin-8 klp5/6p and dam1p contribute to inward pulling forces. Removing these proteins individually led to aberrant metaphase spindle length and chromosome segregation defects. Removing these proteins in antagonistic combination rescued the defective spindle length and, in some combinations, also partially rescued chromosome segregation defects. Our results stress the importance of proper chromosome-to-microtubule attachment over spindle length regulation for proper chromosome segregation.postprin