STUDY QUESTION Is it feasible to disseminate testicular tissue cryopreservation with a standardized protocol through a coordinated network of centers and provide centralized processing/freezing for centers that do not have those capabilities? SUMMARY ANSWER Centralized processing and freezing of testicular tissue from multiple sites is feasible and accelerates recruitment, providing the statistical power to make inferences that may inform fertility preservation practice. WHAT IS KNOWN ALREADY Several centers in the USA and abroad are preserving testicular biopsies for patients who cannot preserve sperm in anticipation that cell- or tissue-based therapies can be used in the future to generate sperm and offspring. STUDY DESIGN, SIZE, DURATION Testicular tissue samples from 189 patients were cryopreserved between January 2011 and November 2018. Medical diagnosis, previous chemotherapy exposure, tissue weight, and presence of germ cells were recorded. PARTICIPANTS/MATERIALS, SETTING, METHODS Human testicular tissue samples were obtained from patients undergoing treatments likely to cause infertility. Twenty five percent of the patient’s tissue was donated to research and 75% was stored for patient’s future use. The tissue was weighed, and research tissue was fixed for histological analysis with Periodic acid-Schiff hematoxylin staining and/or immunofluorescence staining for DEAD-box helicase 4, and/or undifferentiated embryonic cell transcription factor 1. MAIN RESULTS AND THE ROLE OF CHANCE The average age of fertility preservation patients was 7.9 (SD = 5) years and ranged from 5 months to 34 years. The average amount of tissue collected was 411.3 (SD = 837.3) mg and ranged from 14.4 mg—6880.2 mg. Malignancies ( n = 118) were the most common indication for testicular tissue freezing, followed by blood disorders ( n = 45) and other conditions ( n = 26). Thirty nine percent ( n = 74) of patients had initiated their chemotherapy prior to undergoing testicular biopsy. Of the 189 patients recruited to date, 137 have been analyzed for the presence of germ cells and germ cells were confirmed in 132. LIMITATIONS, REASONS FOR CAUTION This is a descriptive study of testicular tissues obtained from patients who were at risk of infertility. The function of spermatogonia in those biopsies could not be tested by transplantation due limited sample size. WIDER IMPLICATIONS OF THE FINDINGS Patients and/or guardians are willing to pursue an experimental fertility preservation procedure when no alternatives are available. Our coordinated network of centers found that many patients request fertility preservation after initiating gonadotoxic therapies. This study demonstrates that undifferentiated...

The significance of the threshold amperage of peripheral nerve stimulation (PNS) for the efficacy and latency of sciatic block is shown in a controlled randomized study of stimulation amplitude. In all cases the block was complete within a short time when the threshold amperage was 0.3 mA or less. Incomplete motor and sensory blocks occurred with higher stimulation amplitudes of 0.5 and 1.0 mA. In view of these results a prospective study of the clinical efficacy of 852 combined sciatic/3-in-1 blocks using prilocaine, and performed by means of peripheral nerve stimulation was carried out. No CNS or cardiovascular complications, no problems resulting from methaemoglobinaemia and, above all, no nervous lesions were observed. The limiting factor for surgery of the lower limb with this method of anaesthesia is the tolerance of the femoral tourniquet which depends mainly on the efficacy of the 3-in-1 block. Ninety-one per cent of the combined blockades were primarily successful when there was no tourniquet at all, and 87% when the tourniquet was placed on the lower leg. In the course of surgery with a femoral pneumatic tourniquet, only 55% of the blocks did not require supplement when 20 ml of 1% prilocaine was used for the 3-in-1 block, while 72% and 74% were efficacious with 30 ml and 35 ml, respectively. The efficacy of the sciatic block proved to be extremely high (> 95%), its success depending on the dosage of the local anaesthetic and correct execution of the peripheral nerve stimulation.

In a pharmacokinetic study of combined sciatic/3-in-1 block for lower limb surgery, the two moderate-acting local anaesthetics prilocaine and mepivacaine were compared. The mean maximum venous plasma concentrations of mepivacaine were more than twice as high as when prilocaine was used as anaesthetic (5.1 micrograms/ml vs. 2.37 micrograms/ml). When used in combination with the former, ornipressin did not reduce plasma concentrations of mepivacaine to values which were below the threshold for toxic symptoms (5-6 mg/ml). The peak plasma concentrations exceeded the threshold of 5 micrograms/ml in four of the nine patients of the mepivacaine group (maximum value 7.21 mg/ml) and in two of the nine patients of the mepivacaine+ornipressin group (maximum value 8.61 micrograms/ml).

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