Clinical comparison between conventional and microdissection testicular sperm extraction for non-obstructive azoospermia: Understanding which treatment works for which patient

Objectives: The superiority of microdissection testicular sperm extraction (mTESE) over conventional TESE (cTESE) for men with non-obstructive azoospermia (NOA) is debated. We aimed to compare the sperm retrieval rate (SRR) of mTESE to cTESE and to identify candidates who would most benefit from mTESE in a cohort of Caucasian-European men with primary couple’s infertility. Material and methods: Data from 49 mTESE and 96 cTESE patients were analysed. We collected demographic and clinical data, serum levels of LH, FSH and total testosterone. Patients with abnormal karyotyping were excluded from analysis. Age was categorized according to the median value of 35 years. FSH values were dichotomized according to multiples of the normal range (N) (N and 1.5 N: 1-18 mIU/mL, and > 18 mIU/mL). Testicular histology was recorded for each patient. Descriptive statistics and logistic regression analyses tested the impact of potential predictors on positive SRR in both groups. Results: No differences were found between groups in terms of clinical and hormonal parameters with the exception of FSH values that were higher in mTESE patients (p = 0.004). SRR were comparable between mTESE and cTESE (49.0% vs. 41.7%, p = 0.40). SRRs were significantly higher after mTESE in patients with Sertoli cell-only syndrome (SCOS) (p = 0.038), in those older than 35 years (p = 0.03) and with FSH >1.5N (p 1.5N (p = 0.018). Moreover, increased FSH levels (p = 0.03) and both SCOS (p = 0.01) and MA histology (p = 0.04) were independent predictors of SRR failure. Conclusions: Microdissection and cTESE showed comparable success rates in our cohort of patients with NOA. mTESE seems beneficial for patients older than 35 years, with high FSH values, or when SCOS can be predicted. Given the high costs associated with the mTESE approach, the identification of candidates most likely to benefit from this procedure is a major clinical need

Transgenic Fatal Familial Insomnia Mice Indicate Prion Infectivity-Independent Mechanisms of Pathogenesis and Phenotypic Expression of Disease

<div><p>Fatal familial insomnia (FFI) and a genetic form of Creutzfeldt-Jakob disease (CJD¹⁷⁸) are clinically different prion disorders linked to the D178N prion protein (PrP) mutation. The disease phenotype is determined by the 129 M/V polymorphism on the mutant allele, which is thought to influence D178N PrP misfolding, leading to the formation of distinctive prion strains with specific neurotoxic properties. However, the mechanism by which misfolded variants of mutant PrP cause different diseases is not known. We generated transgenic (Tg) mice expressing the mouse PrP homolog of the FFI mutation. These mice synthesize a misfolded form of mutant PrP in their brains and develop a neurological illness with severe sleep disruption, highly reminiscent of FFI and different from that of analogously generated Tg(CJD) mice modeling CJD¹⁷⁸. No prion infectivity was detectable in Tg(FFI) and Tg(CJD) brains by bioassay or protein misfolding cyclic amplification, indicating that mutant PrP has disease-encoding properties that do not depend on its ability to propagate its misfolded conformation. Tg(FFI) and Tg(CJD) neurons have different patterns of intracellular PrP accumulation associated with distinct morphological abnormalities of the endoplasmic reticulum and Golgi, suggesting that mutation-specific alterations of secretory transport may contribute to the disease phenotype.</p></div

Tg(FFI) mice show recognition and spatial working memory impairment.

<p>(A) Performance in the novel object recognition task was expressed as a discrimination index (see Experimental Procedures). Histograms indicate the mean ± SEM of 10 non-Tg/<i>Prnp</i>^+/+, 10 non-Tg/<i>Prnp</i>^0/0, and 8 Tg(FFI-26^+/-)/<i>Prnp</i>^0/0 aged 70 days; F_2,25 = 8.3 p = 0.017 by one-way ANOVA; *p < 0.05, **p < 0.01, Tukey’s post hoc test. (B) Histograms represent the mean ± SEM of total errors in the eight-arm radial maze in the first eight trials during 16 days of training, by the same non-Tg/<i>Prnp</i>^0/0 and Tg(FFI-26^+/-)/<i>Prnp</i>^0/0 mice used in A. t₁₆ = 3.0; p = 0.009; **p < 0.01 by Student’s t test. (C) Values are the mean latency (± SEM) to complete the radial maze. F_15,240 = 19; p = 0.03 by one-way ANOVA for repeated measures. *p < 0.05 by Student’s t test.</p

A tetracationic porphyrin with dual anti-prion activity

Prions are deadly infectious agents made of PrPSc, a misfolded variant of the cellular prion protein (PrPC) which self-propagates by inducing misfolding of native PrPC. PrPSc can adopt different pathogenic conformations (prion strains), which can be resistant to potential drugs, or acquire drug resistance, hampering the development of effective therapies. We identified Zn(II)-BnPyP, a tetracationic porphyrin that binds to distinct domains of native PrPC, eliciting a dual anti-prion effect. Zn(II)-BnPyP binding to a C-terminal pocket destabilizes the native PrPC fold, hindering conversion to PrPSc; Zn(II)-BnPyP binding to the flexible N-terminal tail disrupts N-to C-terminal interactions, triggering PrPC endocytosis and lysosomal degradation, thus reducing the substrate for PrPSc generation. Zn(II)-BnPyP inhibits propagation of different prion strains in vitro, in neuronal cells and organotypic brain cultures. These results identify a PrPC-targeting compound with an unprecedented dual mechanism of action which might be exploited to achieve anti-prion effects without engendering drug resistance

Tg(FFI) mice show an altered response to sleep deprivation.

<p>Time course of the loss and recovery of time spent in rapid eye movement (REM) (A) and non-rapid eye movement (NREM) (B) sleep, during and after sleep deprivation. Values were from 8 non-Tg/<i>Prnp</i>^+/+, 10 non-Tg/<i>Prnp</i>^0/0, 9 Tg(FFI-26)/<i>Prnp</i>^0/0 and 8 Tg(FFI-26)/<i>Prnp</i>^+/0. Mice were kept awake during the first 6 h of the light phase (crosshatched bar) by gentle handling, and allowed to sleep freely in the next 18 h. The black bar indicates the dark portion of the light-dark cycle. REM and NREM sleep were calculated hourly for each animal as the difference between the amount of time spent in a given state (REM or NREM sleep) during and after sleep deprivation, and the amount spent in the corresponding hour during baseline conditions (undisturbed). The hour-by-hour differences were then summed to obtain a cumulative curve. Data (means ± SEM) are presented in 2-h intervals. Single symbols: p < 0.05; double symbols: p < 0.01. *, Tg(FFI-26)/<i>Prnp</i>^0/0 vs non-Tg/<i>Prnp</i>^0/0; °, Tg(FFI-26)/<i>Prnp</i>^0/0 vs. non-Tg/<i>Prnp</i>^+/+; §, Tg(FFI-26)/<i>Prnp</i>^0/0 vs. Tg(FFI-26)/<i>Prnp</i>^+/0; #, Tg(FFI-26)/<i>Prnp</i>^+/0 vs. non-Tg/<i>Prnp</i>^+/+. A mixed model analysis of variance for repeated measures was done on 6 h blocks. Between-strains post-hoc comparisons by one-way ANOVA with Bonferroni correction: (panel A) 0–6 h: F_3,101 = 4.98, p = 0.003; 7–12 h: F_3,101 = 5.25, p = 0.002; 13–18 h: F_3,101 = 2.88, p = 0.05; 19–24 h: F_3,101 = 3.30, p = 0.023. (panel B) 0–6 h: F_3,101 = 1.01, p = 0.391; 7–12 h: F_3,101 = 1.78, p = 0.156; 13–18 h: F_3,101 = 3.76, p = 0.013; 19–24 h: F_3,101 = 3.97, p = 0.010.</p