Testicular sperm retrieval: What should we expect from the fresh and subsequent cryopreserved sperm injection?

We sought to compare ICSI outcomes of cycle using fresh versus thawed TESE spermatozoa obtained during the previous fresh TESE. All consecutive couples undergoing ICSI cycles using fresh TESE spermatozoa, followed by ICSI cycle using cryopreserved sperm remaining from the previous fresh TESE procedure were included. Ovarian stimulation (OS)/laboratory variables and cycle outcome were assessed and compared between those utilising fresh versus thawed TESE spermatozoa. Seventy-five couples were evaluated, with no in-between groups differences in OS nor embryological variables. While implantation and LBR per embryo transfer were nonsignificantly higher in the frozen as compared to the fresh TESE, there was a trend towards higher LBRs per patient in the frozen TESE group. The cumulative miscarriage rate (4% versus 14.7%, p < .022 respectively) was significantly lower and the cumulative LBR (34.7% versus 16%, p < .007 respectively) was significantly higher using frozen TESE spermatozoa. Moreover, significantly higher proportion of frozen TESE sperm samples used pentoxifylline to enhance sperm motility. In conclusion, the results of ICSI cycles using frozen TESE spermatozoa are as good, or even better than using fresh TESE spermatozoa. Further studies are required to explore the factors responsible for the improved ICSI outcome, while using frozen versus fresh TESE sperm samples.     

A spectral analysis of the high frequency QRS potentials observed during acute myocardial ischemia in dogs

Electrocardiographic manifestations of transient myocardial ischemia were studied in 10 dogs undergoing occlusion of the left anterior descending coronary artery, using frequency domain analysis of the high frequency QRS potentials. Ischemia was induced via a balloon occluding cuff placed around the coronary artery. Electrocardiographic waveforms recorded before and during the ischemic state were averaged together and then filtered between 150 and 250 Hz to obtain the high frequency QRS complex. The spectral curve in the power spectrum of the high frequency QRS components was calculated using an algorithm for fast Fourier transformation. Balloon inflation was accompanied by morphological changes in the spectral curve in 7 of 10 experiments. Morphological changes, seen as zones of reduced amplitude in the time domain, appear as two separate peaks in the frequency domain. These changes were seen early in ischemia and were not accompanied by significant changes in the QRS complex or ST segment of the conventional surface electrocardiogram. The power spectrum of the high-frequency QRS potentials appears to detect evidence of transient ischemia and may provide useful information regarding the presence of myocardial ischemia.     

Prognostic role of decreased E-cadherin expression in patients with upper tract urothelial carcinoma: a multi-institutional study

Purpose

To assess the role of E-cadherin as prognostic biomarker in upper tract urothelial carcinoma (UTUC) in a large multi-institutional cohort of patients.

Methods

Immunohistochemistry technique was used to evaluate E-cadherin expression in 678 patients with unilateral, sporadic UTUC treated with RNU. E-cadherin expression was considered decreased if 10 % or more cells had decreased expression (<90 %).

Results

Decreased E-cadherin expression was observed in 353 patients (52.1 %) and was associated with advanced pathological stage (P < 0.001), higher grade (P < 0.001), lymph node metastasis (P = 0.006), lymphovascular invasion (P < 0.001), concomitant carcinoma in situ (P < 0.001), multifocality (P = 0.004), tumor necrosis (P = 0.020) and sessile architecture (P < 0.001). Within a median follow-up of 30 months (interquartile range 15–57), 171 patients (25.4 %) experienced disease recurrence and 150 (21.9 %) died from UTUC. In univariable analyses, decreased E-cadherin expression was significantly associated with worse recurrence-free survival (P < 0.001) and cancer-specific survival CSS (P = 0.006); however, in multivariable analyses, it was not (P = 0.74 and 0.84, respectively). The lack of independent prognostic value of E-cadherin remained true in all subgroup analyses.

Conclusion

In UTUC patients treated with RNU, decreased E-cadherin expression is associated with features of biologically and clinically aggressive disease and worse outcome in univariable, but not multivariable, analyses. If E-cadherin’s association with factors of advanced disease is confirmed on UTUC biopsy specimens, it could be used to help in the clinical decision-making regarding kidney-sparing approaches and/or neo-adjuvant chemotherapy.

     

The utility of a single simple question in the evaluation of patients with myasthenia gravis

Introduction: Assessing myasthenia gravis (MG) can be challenging, and multiple scales are available to evaluate disease severity. We evaluated the utility of a single, simple question, as part of the MG evaluation: “What percentage of normal do you feel regarding your MG, 0%–100% normal?” Methods: A retrospective chart review of patients attending the neuromuscular clinic from January 2014 to December 2015 was performed. Responses were correlated with symptoms and signs, the Quantitative Myasthenia Gravis Score (QMGS), the Myasthenia Gravis Impairment Index (MGII), and the 15‐item Myasthenia Gravis Quality of Life scale (MG‐QOL15). Results: The total cohort included 169 patients. The percentage of normal correlated strongly with limb muscle weakness and MG scales, moderately with bulbar and respiratory symptoms, and weakly with ocular manifestations. Discussion: The question, “What percentage of normal do you feel regarding your MG?” is feasible and valid, and can be incorporated easily into routine clinical evaluation. Muscle Nerve 57 : 240–244, 2018     

Long‐lasting pro‐ictogenic effects induced in vivo by rat brain exposure to serum albumin in the absence of concomitant pathology

Purpose: Dysfunction of the blood–brain barrier (BBB) is a common finding during seizures or following epileptogenic brain injuries, and experimentally induced BBB opening promotes seizures both in naive and epileptic animals. Brain albumin extravasation was reported to promote hyperexcitability by inducing astrocytes dysfunction. To provide in vivo evidence for a direct role of extravasated serum albumin in seizures independently on the pathologic context, we did the following: (1) quantified the amount of serum albumin extravasated in the rat brain parenchyma during status epilepticus (SE); (2) reproduced a similar concentration in the hippocampus by intracerebroventricular (i.c.v.) albumin injection in naive rats; (3) measured electroencephalography (EEG) activity in these rats, their susceptibility to kainic acid (KA)–induced seizures, and their hippocampal afterdischarge threshold (ADT). Methods: Brain albumin concentration was measured in the rat hippocampus and other forebrain regions 2 and 24 h after SE by western blot analysis. Brain distribution of serum albumin or fluorescein isothiocyanate (FITC)‐albumin was studied by immunohistochemistry and immunofluorescence, respectively. Naive rats were injected with rat albumin or FITC‐albumin, i.c.v., to mimic the brain concentration attained after SE, or with dextran used as control. Inflammation was evaluated by immunohistochemistry by measuring glial induction of interleukin (IL)‐1β. Western blot analysis was used to measure inward rectifying potassium channel subunit Kir4.1 protein levels in the hippocampus. Seizures were induced in rats by intrahippocampal injection of 80 ng KA and quantified by EEG analysis, 2 or 24 h after rat albumin or dextran administration. ADT was measured by electrical stimulation of the hippocampus 3 months after albumin injection. In these rats, EEG was continuously monitored for 2 weeks to search for spontaneous seizures. Key Findings: The hippocampal serum albumin concentration 24 h post‐SE was 0.76 ± 0.21 μm . Similar concentrations were measured in other forebrain regions, whereas no changes were found in cerebellum. The hippocampal albumin concentration was similarly reproduced in naive rats by i.c.v. administration of 500 μg/4 μl rat albumin: albumin was predominantly detected extracellularly 2 h after injection, whereas at 24 h it was visible inside pyramidal neurons and in only a few scattered chondroitin sulphate proteoglycan (NG2)‐positive cells, but not in glial fibrillary acidic protein (GFAP)‐positive astrocytes or CR‐3 complement receptor (OX‐42)‐positive microglia. The presence of albumin in naive rat hippocampus was associated with induced IL‐1β in GFAP‐positive astrocytes and a concomitant tissue down‐regulation of Kir4.1. Spiking activity was evoked by albumin in the hippocampus lasting for 2 h. When KA was intrahippocampally applied either 2 or 24 h after albumin injection, the number of total interictal spikes in 3 h EEG recording was significantly increased by twofold on average. Three months after albumin injection, neither albumin nor inflammation was detected in brain tissue; at this time, the ADT was reduced by 50% but no spontaneous seizures were observed. Significance: Transient hippocampal exposure to albumin levels similar to those attained after prominent BBB breakdown resulted in increased seizure susceptibility and long‐term reduction in seizure threshold, but it did not evoke spontaneous seizures. These effects may be mediated by albumin‐induced astrocytes dysfunction and the associated induction of proinflammatory molecules.     

Midline thalamic inputs to the amygdala: Ultrastructure and synaptic targets

One of the main subcortical inputs to the basolateral nucleus of the amygdala (BL) originates from a group of dorsal thalamic nuclei located at or near the midline, mainly from the central medial (CMT), and paraventricular (PVT) nuclei. Although similarities among the responsiveness of BL, CMT, and PVT neurons to emotionally arousing stimuli suggest that these thalamic inputs exert a significant influence over BL activity, little is known about the synaptic relationships that mediate these effects. Thus, the present study used Phaseolus vulgaris-leucoagglutinin (PHAL) anterograde tracing and electron microscopy to shed light on the ultrastructural properties and synaptic targets of CMT and PVT axon terminals in the rat BL. Virtually all PHAL-positive CMT and PVT axon terminals formed asymmetric synapses. Although CMT and PVT axon terminals generally contacted dendritic spines, a substantial number ended on dendritic shafts. To determine whether these dendritic shafts belonged to principal or local-circuit cells, calcium/calmodulin-dependent protein kinase II (CAMKIIα) immunoreactivity was used as a selective marker of principal BL neurons. In most cases, dendritic shafts postsynaptic to PHAL-labeled CMT and PVT terminals were immunopositive for CaMKIIα. Overall, these results suggest that CMT and PVT inputs mostly target principal BL neurons such that when CMT or PVT neurons fire, little feed-forward inhibition counters their excitatory influence over principal cells. These results are consistent with the possibility that CMT and PVT inputs constitute major determinants of BL activity.     

Reference values for ultrasonograpy of peripheral nerves

Introduction: High‐resolution ultrasonography (HRU) is a novel method that provides morphological information about peripheral nerves. We aimed to determine reference values for nerve cross‐sectional area (CSA) on HRU. Methods: One hundred healthy volunteers had HRU of median, radial, ulnar, fibular, tibial, sural, and superficial fibular nerves at defined sites. The CSA was measured and the effects of age, gender, and body mass index (BMI) were evaluated. Results: CSA values in healthy subjects are described. CSA is larger in lower limb motor nerves than in sensory nerves at similar sites, and the CSA tends to be symmetrical. The strongest effect on CSA was for age, although gender and BMI had some effects. Conclusions: This study provides normative values for HRU, and it suggests that further research with age‐ and gender‐specific distributions must be a key priority in the development of HRU for use as a diagnostic test for peripheral nerve diseases. Muscle Nerve 53 : 538–544, 2016     

One-to-one neuron–electrode interfacing

The question of neuronal network development and organization is a principle one, which is closely related to aspects of neuronal and network form-function interactions. In-vitro two-dimensional neuronal cultures have proved to be an attractive and successful model for the study of these questions. Research is constraint however by the search for techniques aimed at culturing stable networks, whose electrical activity can be reliably and consistently monitored. A simple approach to form small interconnected neuronal circuits while achieving one-to-one neuron–electrode interfacing is presented. Locust neurons were cultured on a novel bio-chip consisting of carbon-nanotube multi-electrode-arrays. The cells self-organized to position themselves in close proximity to the bio-chip electrodes. The organization of the cells on the electrodes was analyzed using time lapse microscopy, fluorescence imaging and scanning electron microscopy. Electrical recordings from well identified cells is presented and discussed. The unique properties of the bio-chip and the specific neuron–nanotube interactions, together with the use of relatively large insect ganglion cells, allowed long-term stabilization (as long as 10 days) of predefined neural network topology as well as high fidelity electrical recording of individual neuron firing. This novel preparation opens ample opportunity for future investigation into key neurobiological questions and principles.