A short report on current fertility preservation strategies for boys.

BACKGROUND: Advances in cancer treatment have led to improved long-term survival after childhood cancer, but often at a price of impaired future fertility. Fertility preservation (FP) in male children and early adolescents poses unique challenges as efficacy is unproven. OBJECTIVES: To describe characteristics of testicular tissue cryopreservation (TTCP) specimens taken from paediatric and adolescent patients, stratified by age, and prior chemotherapy, if any, and to demonstrate evidence for germ cells. MATERIALS AND METHODS: Retrospective review of gonadal biopsies and clinical records of patients consented into the Royal Children's Hospital FP programme between 1987 and 2015. Tissue was sliced into blocks, with one section sent for histopathology prior to cryopreservation. In boys ≥12 years where spermatogenesis could be expected, a portion of tissue was disaggregated completely to look for mature sperm and if found, additional tissue was dissected and the resulting suspension frozen. RESULTS: Testicular tissue cryopreservation specimens in 44 males (0.3-16.8 years) provided an average of 7.8 slices per patient. All the specimens were taken at the same time as another necessary surgical procedure, under one general anaesthesic. There was only one complication of scrotal wound dehiscence. Seven of the forty-four (15.9%) patients had chemotherapy prior to testicular biopsy, while the rest were chemotherapy naïve. Five of these were prepubertal, and two were pubertal patients. Eleven subjects had tissue dissected with mature sperm found in eight. Of these eight patients where sperm were found, all were pubertal with testicular size of more than 10 mL and showing histological evidence of spermatogenesis. No histologic specimen demonstrated any malignant cells. CONCLUSIONS: Testicular tissue cryopreservation can be performed in young patients without delay, preferably prior to cancer treatment. As testicular tissue contains germ cells from which haploid spermatozoa are ultimately derived, future technologies may allow their utilization for fertility in humans. This may be the only hope for biological offspring in some patients undergoing fertility compromising treatment. Retrieval of mature sperm from some pubertal patients, however, offers realistic hope to these patients of future fertility.

Outcomes from the first assisted reproduction program for HIV-serodiscordant couples in Australia.

OBJECTIVE: To describe the clinical outcomes for all HIV-serodiscordant couples attending an assisted reproduction program. DESIGN, SETTING AND PARTICIPANTS: Retrospective review of demographic, clinical and outcome data for all HIV-serodiscordant couples who attended an assisted reproduction program at a tertiary hospital in Melbourne, between its commencement in 2003 and June 2010. MAIN OUTCOME MEASURES: Pregnancies, miscarriages, births, HIV transmission to the HIV-negative partner, semen quality and detection of HIV (HIV RNA and HIV DNA) in semen. RESULTS: As of June 2010, 39 HIV-positive clients had proceeded to assisted reproduction after the initial consultation in the program. There were 162 completed cycles, with 26 pregnancies (clinical pregnancy rate per cycle, 16.2% for HIV-positive men with an HIV-negative partner, and 15.4% for HIV-positive women). Of all 222 tested semen samples, 18 (8%) had HIV RNA detected despite these men receiving antiretroviral therapy and having an undetectable HIV viral load in plasma. Sperm velocity was significantly lower in HIV-positive clients receiving combination antiretroviral therapy than in a control group of recipient-recruited sperm donors (P = 0.01); there were no other significant differences in sperm quality between the two groups. No HIV transmission to babies or HIV-negative partners occurred. CONCLUSION: Our findings show detectable HIV in 8% of semen samples from men with an undetectable HIV viral load in plasma, but confirm the safety of assisted reproduction for HIV-serodiscordant couples within a program with strict protocols for HIV treatment and testing of all semen before use.

Lung Cancer Screening Reveals a Nonspiculated Nodule: Histoplasmosis.

We report the case of a middle aged man, 43-pack-years active smoker, who upon radiographic screening for lung cancer found to have a non-spiculated lung nodule concerning for malignancy. Histologic evaluation of this lesion turned out to be a nodule due to Histoplasma capsulatum.

An Unusual MRI Appearance of Osseous Metastases.

Bone metastases can present in a wide variety of appearances across all imaging modalities. We present a unique appearance of a distal femoral metastasis in a patient who initially complained of knee pain. The radiographic and CT findings were initially suspicious for calcium pyrophosphate deposition (CPPD) arthropathy; however, an MRI demonstrated multiple lesions with a lamellated appearance confirmed on biopsy to be metastatic disease. This unusual lamellated appearance has not been previously described. We present this case to help distinguish this entity radiographically and better classify this finding as a manifestation of metastatic disease.

Analysis of HIV-1 viral load in seminal plasma samples.

BACKGROUND: With HIV-1-infected individuals now facing the prospect of relatively long and healthy lives, many discordant couples (where the male is HIV-1 seropositive) are seeking to have children. To assist reducing the risks of heterosexual and subsequent vertical transmission in this situation, quantification of HIV-1 viral load in seminal plasma may be effective as one of several measures to reduce the risk of infecting the mother during insemination, potentially providing a better indication of infectivity than blood plasma analysis. OBJECTIVE(S): To modify existing molecular methods for the purpose of analysing HIV-1 viral load in seminal plasma. METHODS: Two commercial assays for HIV-1 RNA quantification were used to assess their sensitivity, specificity and precision for quantification of seminal plasma samples. Seminal plasma samples were prepared with an additional centrifugation step to aid removal of inhibitors to molecular assays. RESULTS: Seminal plasma samples exhibited specificity of >95%, equivalent to that reported by the manufacturers of the commercial assays. With additional centrifugation, complete inhibition of 2/19 (10%) seminal plasma samples was observed using the RT-PCR assay, and inhibition was not apparent in the bDNA assay. Quantification of HIV-1 RNA in seminal plasma samples in both assays was equivalent to that observed in plasma samples and did not appear to be affected by the additional centrifugation step. CONCLUSION: Minor modification of the RT-PCR assay procedure by additional centrifugation of seminal plasma improved the sensitivity of the assay. Inhibition was not apparent with the bDNA assay.

Semen analysis: its place in modern reproductive medical practice.

Semen analysis is the most important laboratory investigation for men when assessing the infertile couple. Advances in in vitro fertilisation (IVF) techniques, particularly intracytoplasmic sperm injection (ICSI) involving the direct injection of a single spermatozoon into an egg, have not diminished the role of semen analysis in modern reproductive practice. Semen analysis is the most basic laboratory investigation undertaken and is descriptive in terms of semen volume, appearance, viscosity, sperm concentration, sperm motility and morphology. Since the results are used by clinicians to choose appropriate treatment options, a reliable service is imperative. It is crucial that the laboratory is experienced in the performance of semen analyses to ensure an accurate result. To ensure a quality semen analysis service, laboratories must participate in internal and external quality assurance activities, incorporate rigorous training protocols for technical staff and use reliable procedures. The World Health Organization laboratory manual for the examination of human semen and sperm cervical mucous interaction, clearly describes the variables that need to be assessed and the methods of analysis and quality assurance to be used.

Comparison of sperm counts in two groups of men presenting for infertility investigations 20 years apart.

Aims: To compare sperm counts for two groups of men who had presented for infertility investigations approximately 20 years apart. Methods: The study compared results for 309 men tested between 1977 and 1981 with those of 559 men tested between 1997 and 1998 using identical methodology. In order to approximate the normal population, only those men with counts above 5 million/mL were included in the final analysis. Bias, due to repeated testing after an initial abnormal result, was minimized by including only the patient's first test results. In addition, to allow for time-dependent changes in the requirements for semen samples, results were included only if a complete sample was produced by masturbation after 3-5 days abstinence. Results: There was a small, but statistically significant drop in ejaculate volume (3.9-3.6 mL, P = 0.015) and a significant increase in the patient's mean age (32.18 vs 35.08, P < 0.001). Both groups had median abstinence of 3 days and no difference in sperm counts with a mean (median) count for the early group of 87.9 (75) versus 92.0 (76) for the recent group (P > 0.80). The significant drop in ejaculate volume was not reflected in a difference (P = 0.45) in total sperm numbers in the ejaculate with 320.7 (255) versus 313.1 (234). Conclusion: This study found no evidence of a decrease in sperm counts or total sperm output in men (excluding those with severe oligospermia) presenting for infertility investigations in Melbourne, Australia, over the last two decades of the twentieth century. (Reprod Med Biol 2004; 3: 211-216).

Etiology of sperm immunity in women.

Sperm immunity in females can reduce the likelihood of natural conception, and sperm antibodies from female sera have been shown to inhibit IVF in humans and in several animal models. The etiology of sperm immunity in human females is unknown, but several possible mechanisms have been proposed, including cross-reactivity with microbial antigens and interferon gamma-mediated potentiation of the antisperm immune response in women whose male partners have sperm autoantibodies in their semen. This article reviews these ideas and postulates a novel hypothesis based on the potential for the generation of anti-idiotype antibodies in women whose partners have sperm antibodies in their semen.

A.R.T. and history, 1678-1978.

Louise Brown, the first baby conceived after IVF, was born on 25 July 1978 and turned 27 last year. From one perspective, her birth can be seen as the culmination of 300 years of medical and scientific investigation aimed at understanding the fascinating process of reproduction. This essay was written as a tribute to mark the unique contribution to assisted reproductive technology (ART) which resulted from the collaboration of a scientist, Bob Edwards, and a clinician, Patrick Steptoe, who pioneered the successful clinical use of IVF. This article was not intended to be a conventional history of science, but instead has primarily focused on those early discoveries which in the author's opinion were critical to our current understanding of mammalian reproduction. There are some digressions and many omissions necessitated by attempting to cover 300 years in a relatively short essay. In particular, there is no mention of endocrinology because this area has been covered in numerous reviews and books. The main sources of historical information for this article were the authoritative books of Professor Cole (1930), Dr Elizabeth Gasking (1967), Professor John Farley (1982), Dr Fielding H. Garrison (1929) and the Philosophical Transactions of The Royal Society or Letters collated from the latter.

Association between sperm autoantibodies and enhanced embryo implantation rates during in vitro fertilization.

Previous investigations have demonstrated the propensity of strong IgA-class sperm autoantibodies to impede fertilization. However, because there has not been a general consensus on this issue, the aim of this retrospective analysis was to focus on the effects of different levels of IgA-class antibodies on each stage of the IVF procedure. This study has confirmed that high level IgA class antibodies significantly reduce fertilization rates but, unexpectedly, also has shown a very significant improvement in embryo implantation rates in patients with weak to moderate antibody levels. Interlaboratory prospective collaborative studies are being planned to test this preliminary observation more stringently.

Sperm immobilizing properties of lemon juice.

The aim of this study was to evaluate the sperm-immobilizing properties of lemon juice to determine if they are consistent with its traditional contraceptive use. It was found that lemon juice supernatant (LJS) has high osmolality (550-60 mOsm) and low pH (2.2-2.6) and that addition of LJS to semen to give a final concentration of 20% v/v reduced the pH from around 8.4 to 4.1. This acidification was associated with irreversible cessation of all sperm movements within 1 minute. In conclusion, lemon juice should be further evaluated for acceptability, safety, and efficacy as a topical vaginal contraceptive agent.

Recovery of human sperm motility and ability to interact with the human zona pellucida after more than 28 years of storage in liquid nitrogen.

Six donor semen samples were evaluated after 28 years cryopreservation in liquid nitrogen. The results showed that the samples retained good postthaw motility recovery and normal levels of binding to the human zona pellucida and that four of the five samples tested also gave normal levels of zona-induced acrosome reaction. In conclusion, human sperm can survive very long-term storage, which is pertinent information for clinicians referring boys and young men for sperm banking before chemotherapy.

Flow cytometry and microscopic acridine orange test: relationship with standard semen analysis.

Improved prediction of male fertility requires advances in semen analysis. This study examined the reproducibility and independence of the flow cytometry acridine orange test (FCM-AOT) of sperm chromatin integrity as an assessment of semen quality. The study found that FCM-AOT results are not significantly affected by up to 6 h delay in semen preparation (n = 9) or contamination of semen with moderate concentrations of bacteria (<10(8)/ml E. coli or Staph. epidermidis, n = 14). The variation of replicate measurements within samples was low (%Abnormal alpha(t): SD = 1.4, 95%CI = 4.6, n = 25) and different samples from the same men were mostly within the range of measurement error (n = 35). FCM-AOT variables, in particular %Abnormal alpha(t), displayed significant correlations with motility (r = -0.557), vitality (r = -0.469) and morphology (r = -0.464, n = 201), which are similar in magnitude to those existing between the standard semen variables. Surprisingly, no correlation was found between %Abnormal alpha(t) and the microscopic acridine orange test (M-AOT) (n = 185), suggesting the FCM results are sensitive to a different aspect of sperm quality. In summary, this study confirms that although not totally independent of standard semen analysis or the M-AOT, it is found to be a robust, sensitive and reproducible measure of semen quality, representative of the individual.