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Biomedical subjects

R Gosden

Publications and source records attributed to R Gosden.

At least 19 recordsLinked to original sources
PubMed2006

Cryopreservation of murine embryos, human spermatozoa and embryonic stem cells using a liquid nitrogen-free, controlled rate freezer.

A Stirling Cycle Cryocooler has been developed as an alternative to conventional liquid nitrogen controlled rate freezers. Unlike liquid nitrogen systems, the Stirling Cycle freezer does not pose a contamination risk, can be used in sterile conditions and has no need for a constant supply of cryogen. Three types of samples from two species (murine embryos, human spermatozoa and embryonic stem cells), each requiring different cooling protocols, were cryopreserved in the Stirling Cycle freezer. For comparison, cells were also frozen in a conventional liquid nitrogen controlled rate freezer. Upon thawing, the rates of survival of viable cells were generally greater than 50% for mouse embryos and human embryonic stem cells, based on morphology (mouse embryos) and staining and colony formation (human embryonic stem cells). Survival rates of human spermatozoa frozen in the Stirling Cycle freezer, based on motility and dead cell staining, were similar to those of samples frozen in a conventional controlled rate freezer using liquid nitrogen.

Animals↗
PubMed2004

Assessment of ovarian reserve. Ovarian biopsy is not a valid method for the prediction of ovarian reserve.

The evaluation of ovarian reserve, often critical for the elderly infertile woman, is notoriously difficult and inaccurate. The place of ovarian biopsy in this evaluation has been hotly disputed for three decades, but not resolved. To examine the feasibility of ovarian biopsy for this purpose, a project was designed to estimate the total number of oocytes in a human ovary and investigate whether any biopsy regimen is representative of the follicular reserve in an individual. Ovaries removed from patients of reproductive age during operations not involving ovarian pathology were utilized to count the number and type of follicles found in multiple biopsies of 2 and 5 mm and in the whole ovary. Representative results taking into account the total number of follicles found in the whole ovary showed that predicted values based on the biopsies were extremely varied. We concluded that due to the huge variation in the distribution of follicles across the surface of the ovary, there is no place for this procedure in clinical evaluation of reproductive ageing in the individual patient.

Adult↗
PubMed2004

Spindles, mitochondria and redox potential in ageing oocytes.

Studies of human oocytes obtained from women of advanced reproductive age revealed that spindles are frequently aberrant, with chromosomes sometimes failing to align properly at the equator during meiosis I and II. Chromosomal analyses of donated and spare human oocytes and cytogenetic and molecular studies on the origin of trisomies collectively suggest that errors in chromosome segregation during oogenesis increase with advancing maternal age and as the menopause approaches. Disturbances in the fidelity of chromosome segregation, especially at anaphase I, leading to aneuploidy are prime causes of reduced developmental competence of embryos in assisted reproduction, as well as being responsible for the genesis of genetic disease. This review provides an overview of spindle formation and chromosome behaviour in mammalian oocytes. Evidence of a link between abnormal mitochondrial function in oocytes and somatic follicular cells, and finally disturbances in chromosome cohesion and segregation, and cell cycle control in aged mammalian oocytes, are also discussed.

Aneuploidy↗
PubMed2002

Isolation, characterization and expression of the human Factor In the Germline alpha (FIGLA) gene in ovarian follicles and oocytes.

The Factor In the Germline alpha (FIGalpha) transcription factor regulates expression of the zona pellucida proteins ZP1, ZP2 and ZP3 and is essential for folliculogenesis in the mouse. Using the published mouse Figla sequence, BLAST searches identified a human chromosome 2 BAC clone with high sequence identity. Using PCR primers derived from this clone, amplicons derived from ovarian follicles and mature oocytes revealed 100% identity with the appropriate human BAC clone, the expected homology with the mouse Figla gene sequence, and homology on translation with the FIGalpha protein identified in the Japanese rice fish, medaka (Oryzias latipes). PCR expression profiling of this transcript revealed FIGLA mRNA expression in cDNA derived from ovarian follicles (5/5 samples from the primordial through to the secondary stage) mature oocytes (6/9 samples), and less frequently in preimplantation embryos (2/7 samples). Subsequent BLAST searches revealed the predicted full length coding sequence of the human FIGalpha protein which demonstrates 68 and 25% similarity overall to mouse and medaka proteins respectively, with 96 and 57% identity respectively within the basic helix-loop-helix region. This confirms our identification of the human homologue for this gene which maps to chromosome 2p12. Further work is required to understand its role in normal human oocyte development and the potential involvement in human infertility.

Alternative Splicing↗
PubMed2002

Spermatogenesis and steroidogenesis in mouse, hamster and monkey testicular tissue after cryopreservation and heterotopic grafting to castrated hosts.

Retrieval, extracorporal storage and autotransplantation of testicular tissue could become an important strategy for preserving male gonadal function. The present study used syngeneic and immunodeficient nude mice as hosts, and immature and adult mice, neonatal and adult photoregressed Djungarian hamsters and neonatal marmosets to identify the potential of testicular tissue grafting to maintain the morphological and functional integrity of the testis. Testicular tissue was grafted s.c. either as fresh tissue or after cryopreservation into adult, orchidectomized hosts. The mice that received rodent testis tissue were autopsied 50 days later, and blood samples were collected. Sixty-five per cent of mouse isografts contained morphologically normal testicular tissue and seminiferous tubules with some degree of spermatogenic recovery. Mature spermatozoa were recovered after enzymatic disaggregation. Although the recovery of spermatogenesis was limited in adult mouse and hamster tissue, complete spermatogenesis was observed in grafts from immature rodents. Testicular tissue from neonatal marmosets developed up to the stage of spermatocytes at day 135 after xenografting. Androgen concentrations were comparable in intact control mice and in mice receiving fresh mouse and hamster grafts, slightly lower in mice receiving cryopreserved grafts and adult photoregressed hamster tissue, and low in castrated control mice and in mice receiving marmoset tissue. These results show that isografts and xenografts of immature and adult testicular tissue become functionally active as a s.c. graft in the mouse and that this approach might be useful in combination with cryopreservation as a tool for storage and activation of the male germ line and androgen replacement therapy in patients.

Animals↗
PubMed2001

Ovarian tissue harvested from lymphoma patients to preserve fertility may be safe for autotransplantation.

BACKGROUND: Ovarian failure is a common sequel to chemo/radiotherapy in patients successfully treated for cancer. Harvesting, cryopreserving and subsequently re-implanting ovarian cortical grafts can be used to re-establish reproductive potential in women with cancer. The safety issue, however, is of great concern because residual disease in autografted ovarian tissues might cause recrudescence of disease. METHODS: A total of 30 non-obese diabetic severe combined immunodeficient (NOD/LtSz-SCID) mice were individually xenografted s.c. with frozen-thawed ovarian tissue from 18 patients with lymphoma [13 Hodgkin's lymphoma (HL) and 5 non-Hodgkin's lymphoma (NHL)]. The animals were autopsied at 16 weeks, or earlier if cachectic. The xenograft, liver, spleen, sternum, para-aortic lymph nodes and thymus were prepared for histology, immunohistochemistry and human DNA microsatellite analysis. RESULTS: None of the animals grafted with ovarian tissue from lymphoma patients developed disease. However, all 3 animals grafted with lymph node tissue from an NHL patient developed B-cell lymphomas that were confirmed as human in origin by DNA microsatellite analysis. CONCLUSION: Ovarian tissue harvested before high-dose chemotherapy for HL or NHL may not carry a risk of disease transmission by autotransplantation, although the possibility is difficult to exclude completely.

Adult↗
PubMed1999

Early psychosis.

Explore the source record for details and available documents.

Humans↗
PubMed1999

Prepsychotic treatment for schizophrenia: preventive medicine, social control, or drug marketing strategy?

The definition of schizophrenia is currently being extended to include a "prepsychotic" phase. Prepsychosis detection and intervention programs have already been established in Australia. These are intended to identify people "at-risk" for schizophrenia and treat them to prevent their transition into psychosis. However, analysis of leading research in this field shows high levels of arbitrariness in the selection of diagnostic indicators and a lack of convincing evidence about the efficacy of treatments. The favored prophylactic treatment is atypical neuroleptic medication, and sponsorship of research is providing manufacturers of these drugs with a ubiquitous presence in the field. Many risks are associated with atypical neuroleptics and adverse reactions include psychosis. Taken together these factors suggest that prepsychotic intervention may be more concerned with expanding the market for atypical neuroleptics than with preventing schizophrenia.

Adolescent↗
PubMed1998

The molecular basis of oocyte growth and development.

The mammalian oocyte is formed early in foetal life and may remain suspended at diplotene of the first meiotic prophase until several decades later. When, following follicular recruitment, the oocyte enters a growth phase where it increases in volume, and undergoes replication and redistribution of the cytoplasmic organelles. These modifications, including secretion of the zona pellucida, reflect a period of intensive RNA synthesis as the oocyte grows and accumulates the molecular program for embryogenesis. Throughout oocyte development follicle cell support is fundamental to provide the germ line cell with nutrients and growth regulators to ensure progression through the protracted growth phase. Conversely, the oocyte actively promotes growth and differentiation of the follicular cells. Finally, re-initiation of meiosis in mature oocytes results in the production of haploid gametes which are capable of supporting early embryo development. Our present knowledge of the molecular biology of mammalian oogenesis is discussed in this review.

Animals↗
PubMed1998

Conservation of the follicular population in irradiated rats by the cryopreservation and orthotopic autografting of ovarian tissue.

AIM: To preserve the follicular population in rats prior to abdominal irradiation by the orthotopic autografting of ovarian cortical slices (frozen/thawed) in one of two cryoprotective agents (CPAs). The proportion of follicular survival will be quantified at autopsy and a comparison will be made of the two CPAs. METHOD: Thirty Wistar rats, aged 38-39 days, were unilaterally ovariectomised. The ovaries of twenty animals were slow-cooled in either 1.5 M dimethylsulphoxide (DMSO; group B) or 1.5 M ethylene glycol (EG; group C). The ovaries of the remaining ten animals (group A) were fixed and histologically examined to determine a fresh follicle count. The contralateral ovary was irradiated with a single dose of 2.5 Gy. Frozen tissue (from groups B and C) was thawed rapidly and autografted into the irradiated ovarian bursa. All animals were mated eighteen days after grafting and were sacrificed eighteen days later. At autopsy, the number of embryos was counted and the ovarian weights recorded, before the tissue was prepared histologically for quantification of follicular survival. RESULTS: There was no significant difference in the pregnancy rate in the three groups and, although there was a higher number of embryos in the irradiated control group A compared with groups B and C, the difference was not significant. The follicle count in groups B and C was significantly higher than in group A. In group A, the percentage of growing follicles was higher than in groups B and C and the fresh ovary. CONCLUSIONS: Autografting frozen/thawed ovarian tissue preserved a proportion of the follicular population in irradiated rats. There was no significant difference in the numbers of follicles surviving cryopreservation in the two CPAs. Since the irradiated rats were not totally sterilised, we cannot conclude that cryopreserved grafts can restore fertility.

Animals↗
PubMed1997

Growth and development of the mammalian oocyte.

The oocyte is not only the rarest and the largest cell in the body, but it also has one of the most remarkable life histories. Formed in the fetal ovary and suspended at diplotene of meiosis, it may wait for years before beginning to grow, and not until this process is complete can it resume meiosis and undergo fertilisation. Major changes in the number, morphology and distribution of cytoplasmic organelles occur during growth, and a molecular program for embryogenesis is formed. Specific yolk proteins are absent and much of the RNA and some of the protein are degraded by the cleavage stage. The zona pellucida has been intensively studied, but knowledge of oocyte-specific genes is otherwise surprisingly patchy given the significance of this cell type and the expansion of reproductive technology. Finally, it is now clear that oocytes are not mere passengers which depend on granulosa cells for nutrition and regulation but actively promote the growth and differentiation of their follicles.

Aging↗
PubMed1997

Programmed cell death in the reproductive system.

The reproductive system presents some of the best examples of programmed cell death, which is to be expected considering the dramatic cycles of tissue growth and regression in females. Hormones from the pituitary gland, gonads and uterus are responsible for coordinating cycles in which the preservation of cell survival and inhibition of apoptosis are important. In the ovary, atresia regulates the size of the follicle cohort for ovulation and is an archetype of apoptosis as induced by hormone withdrawal. The fate of an antral follicle--growth or atresia--is determined by circulating levels of gonadotrophins, and follicle-stimulating hormone (FSH) in particular. At the end of a menstrual cycle or pregnancy or lactation, hormone withdrawal triggers cell death and tissue remodelling and initiates a fresh cycle. In the endometrium, breast and prostate gland, steroid hormones are the principal survival factors and castration triggers regression of responsive tissues, which is sometimes decisive in the fight against disease. But while the primary trigger of cell death varies between tissues, underlying cellular mechanisms are more conservative and cell death/survival genes, such as bcl-2, bax and others that are expressed in other tissues, play important roles in the reproductive system too.

Apoptosis↗
PubMed1996

[Follicle freezing and autografting. A new method of medically assisted procreation?].

Although frozen human follicles were shown to be viable after thawing, more than forty years, the low yield of primordial follicles after cryopreservation has greatly limited research in this area. The aim of freezing follicles is to stock a patient's ovocytes before potentially destructive therapy for cancer then to reimplant the follicular tissue after successful treatment. The patient's ovulatory capacity would then theoretically be restored, allowing natural or possibly in vitro fertilization. As have other teams, we recently renewed work in cryopreservation of follicles in experimental models in search for a better cryopreservation agent and have had encouraging results suggesting that primordial human follicles or ovarian fragments can be grafted successfully after cryopreservation. Questions still under study include the optimal site for implantation, the survival time for grafts and the quantity of follicles needed to achieve pregnancy. Work on large animal models or xenografts of human tissues on immunotolerant animals may provide further insight. The possibilities of grafting primordial follicles, either alone or within ovarian tissue, may raise hope for many women, although new questions such as an age limit for reimplantation and use of allografts requiring preparation with major immunosuppressive therapy require further debate.

Animals↗
PubMed1996

The social construction of drug debates.

Drug debates normally proceed without scrutiny of why the particular issues being debated are considered the crucial issues. One plausible influence on the terms of debates is the interest groups involved. Four drug debates are addressed in this paper: neuroleptics, drugs in sport, analgesics and marijuana. The key interest groups and their likely influence on the terms of debates are examined. The implication of this analysis is that more attention should be directed to the terms of drug debates rather than just arguing within those terms.

Analgesics↗
PubMed1996

Low temperature storage and grafting of human ovarian tissue.

Ovarian tissue storage at low temperatures is a promising new method for protecting young cancer patients from the sterilizing effects of chemotherapy and/or radiotherapy. Tissue can be stored and returned to the body in due course as a thin cortical graft since the primordial follicles are distributed peripherally and are relatively resistant to ischaemia. Slices of tissue donated by healthy patients were trimmed to a uniform size and preserved by slow freezing to liquid nitrogen temperatures for up to 2 months in one of the following cryoprotectants: dimethylsulphoxide, ethylene glycol, glycerol, propylene glycol. Their viability was assessed by counting follicles in histological sections 18 days after grafting under the kidney capsules of severe combined immunodeficiency (SCID) mice, and the results were expressed as percentages of the numbers in comparable pieces of ungrafted tissue. While only 10% of the total number of follicles was found in the glycerol group compared to controls, significantly higher percentages (44-84%) survived cryopreservation in the other media. The tissues were sterile when frozen and thawed without a cryoprotectant. These results suggest that if comparable results could be achieved by autografting, a patient's fertility should be safeguarded from cytotoxic treatment.

Adolescent↗