Guidelines for counselling in infertility: outline version.

The Guidelines for Counselling in Infertility describe the purpose, objectives, typical issues and communication skills involved in providing psychosocial care to individuals using fertility services. The Guidelines are presented in six sections. The first section describes how infertility consultations differ from other medical consultations in obstetrics and gynaecology, whereas the second section addresses fundamental issues in counselling, such as what is counselling in infertility, who should counsel and who is likely to need counselling. Section 3 focuses on how to integrate patient-centred care and counselling into routine medical treatment and section 4 highlights some of the special situations which can provoke the need for counselling (e.g. facing the end of treatment, sexual problems). Section 5 deals exclusively with third party reproduction and the psychosocial implications of gamete donation, surrogacy and adoption for heterosexual and gay couples and single women without partners. The final section of the Guidelines is concerned with psychosocial services that can be used to supplement counselling services in fertility clinics: written psychosocial information, telephone counselling, self-help groups and professionally facilitated group work. This paper summarizes the different sections of the Guidelines and describes how to obtain the complete text of the Guidelines for Counselling in Infertility.

Physical factors are involved in the destruction of embryos and oocytes during freezing and thawing procedures.

Observations made during the freezing and thawing of mouse and human oocytes and mouse embryos with the cryomicroscope suggest that physical factors as well as physicochemical factors may play a role in the development of lethal damage upon thawing. The point of contact with the approaching ice front may predispose that area to the appearance of future cytoplasmic blebbing. The ice front distorts the oocyte and this distortion remains during its subsequent thermal history and is unrelated to desiccation distortion. Ice initiates the formation of both intra- and extracellular gas bubbles which are apparent upon thawing; with the progression of the thawing process they can be seen to grow in volume. Growth of these bubbles can give rise to expanding vesicles which can totally destroy an embryo. The consequences of these physical factors for the successful cryopreservation of oocytes, embryos, tissues and organs are discussed.

Morphology and X-ray microprobe analysis of spermatozoa from fertile and infertile men in in vitro fertilization.

This paper reports differences observed in the elemental content of fertile and infertile human spermatozoa used in an in vitro fertilization (IVF) program. "Fertile" and "infertile" were designated by the successful penetration or failure to penetrate an oocyte in vitro. We report morphological and morphometric differences which, together with elemental changes, may be causes of infertility in apparently normal spermatozoa. There were significant differences (P less than 0.05) in sodium and chlorine concentrations between fertile and infertile samples and there was more chlorine than could be accounted for as sodium chloride. Many spermatozoa showed particles adhering to tails, with a higher incidence of "contamination" in the infertile spermatozoa. There were significant differences in both shapes of heads and lengths of tails between fertile and infertile spermatozoa.

X-ray microanalysis of proximal and distal tubule cells in the mouse kidney, and the influence of cadmium on the concentration of natural intracellular elements.

Quantitative X-ray microanalysis of frozen freeze-dried sections of mouse cortex have been used to determine the concentrations of Na, Mg, P, S, Cl, K, Ca and Cd in normal mice and those subjected t 0.7 mumol of cadmium chloride in two subcutaneous injections. These injections result in tissue levels of approximately 100 mg Cd/kg dry weight (less than 1 mM) in whole kidney when analysed by atomic absorption spectrometry. There were distinct and characteristic differences -- 'fingerprints' -- in the elemental composition of both cytoplasm and mitochondria in proximal and distal tubules of normal mice that were distributed by the cadmium treatment. The most significant effect of the cadmium injections was a highly significant increase in the sulphur content of the cytoplasm and mitochondria of distal tubules and a loss in concentration of Mg, P, Cl, K, and particularly Na, from the mitochondria. These results are discussed in the light of current concepts of metallothionein induction (metallothionein is a sulphur-rich protein that acts to bind, amongst other metals, cadmium) and the lack of damage observed in the distal tubules.

X-ray microanalysis of HeLa S3 cells. II. Analysis of elemental levels during the cell cycle.

HeLa S3 cells were synchronized using hydroxyurea. Cryoultramicrotomy and X-ray microanalysis were used to study changes occurring in concentrations of elements during the cell cycle of the synchronized cells. Three subcellular compartments were studied : cytoplasm, nucleus and nucleolus. Potassium concentrations showed little fluctuation in all of the cell compartments during the cell cycle. Sodium concentrations increased during S. and M phases, returning to lower levels in the G1 phase. Chlorine concentrations were highest during the S and G2 phases. At all stages of the cell cycle respective concentrations of potassium, sodium, sulphur and chlorine were similar in the cytoplasm and nucleus. Concentrations of phosphorus increased in the nucleus during S, G2 and M, and also showed fluctuations in the nucleolus during the cycle; these were not seen in the cytoplasm. In S, M and M/G1 sodium concentrations were highest in the nucleolus compared with the other compartments. In the cytoplasm these changes resulted in an increase in total monovalent cation concentration (i.e. sodium + potassium) during S, G2 and M, which returned to base levels after mitosis. This increase in monovalent cation concentration is due almost entirely to the increase in sodium, with little change occurring in the concentration of potassium.

X-ray microanalysis of HeLa S3 Cells. I. Instrumental calibration and analysis of randomly growing cultures.

Cryo-ultramicrotomy and X-ray microanalysis were used to study the elemental composition of HeLa S3 cells. Quantitation was achieved by reference to elemental standards of known concentration made up in 25% gelatin. Analysis of standards showed linear calibration for each of the elements studied: Na, P, S, Cl, K. Standardization was validated by comparing flame-photometric analysis of gelatin containing sodium potassium tartrate with that of X-ray microanalysis. Freeze-dried sections of cells showed good morphology and analysis of whole sections of the cells showed that K/Na varied in individual cells. Low K/Na could not be ascribed to cell damage or to the sequestering of Na in any particular subcompartment of the cells. Treatment with ouabain caused changes in levels of all the elements studied and resulted in a low K/Na ratio in all cells.

Ionic gradients within mantle-collar epithelial cells of the land snail Otala lactea.

Aestivating Otala lactea have been shown to reduce the rate of evaporative water loss from the cells of the mantle-collar epithelium to a level comparable to that of an insect. X-ray microanalyses of ultrathin frozen sections from aestivating and non-aestivating snails have shown gradients of chloride and potassium ions in the apical microvillus region of the regulating mantle collar epithelium. The greatest difference in osmotic concentration occurs in the apical 2 micron of the cell. There appears to be a barrier at that level that prevents water being mobilised from the underlying tissues. Methods of presenting data generated by X-ray microanalysis are also discussed.