We investigated the effects of BTX-A on visceral afferent nerve transmission by measuring bladder tissue NGF levels in patients with neurogenic detrusor overactivity before and after intravesical treatment with BTX-A. We also compared the bladder tissue NGF content with clinical and urodynamic data. A total of 23 patients underwent clinical evaluation and urodynamics with detection of the UDC threshold, maximum pressure and maximum cystometric capacity before, and at the 1 and 3-month followups. Endoscopic bladder wall biopsies were also obtained at the same time points. NGF levels were measured in tissue homogenate by enzyme-linked immunosorbent assay (Promega, Madison, Wisconsin). At 1 and 3 months mean catheterization and incontinent episodes were significantly decreased (p <0.05 and <0.001, respectively). On urodynamics we detected a significant increase in the UDC threshold and maximum cystometric capacity, and a significant decrease in UDC maximum pressure at the 1 and 3-month follow-ups compared to baseline (each p <0.001). At the same time points we detected a significant decrease in NGF bladder tissue content (each p <0.02). BTX-A intravesical treatment induces a state of NGF deprivation in bladder tissue that persists at least up to 3 months. As caused by BTX-A, the decrease in acetylcholine release at the presynaptic level may induce a decrease in detrusor contractility and in NGF production by the detrusor muscle. Alternatively BTX-A can decrease the bladder level of neurotransmitters that normally modulate NGF production and release.

Using the rabbit as an animal model, this study evaluated the long-term effect of silver nanoparticles (NPs) administered intravenously (0.6 mg/kg bw) on reproductive activity and sperm quality. Semen analysis was performed by optical microscopy and sperm motility evaluation by computer assisted sperm analyzer (CASA). Mitochondria oxygen consumption, light and transmission electron microscopy of rabbit testis and ejaculated sperm were also carried out. Throughout the experiment NP-treated rabbits showed higher seminal reactive oxygen species (ROS), less motile sperm, and lower curvilinear velocity and oxygen consumption than control animals. In contrast, libido, serum testosterone, sperm concentration, and semen volume were hardly affected by NPs. Transmission electron microscopy analysis did not show any evident morphological damage in testes; however, Ag NPs are visible in spermatids and ejaculated sperm. These preliminary results show that Ag NPs can reach the testes, compromising sperm motility, sperm speed, and acrosome and mitochondria shape and function.

Activation of brain mitochondrial phospholipase(s) A₂ (PLA₂) might contribute to cell damage and be involved in neurodegeneration. Despite the potential importance of the phenomenon, the number, identities, and properties of these enzymes are still unknown. Here, we demonstrate that isolated mitochondria from rat brain cortex, incubated in the absence of respiratory substrates, release a Ca²⁺-dependent PLA₂ having biochemical properties characteristic to secreted PLA₂ (sPLA₂) and immunoreacting with the antibody raised against recombinant type IIA sPLA₂ (sPLA₂-IIA). Under identical conditions, no release of fumarase in the extramitochondrial medium was observed. The release of sPLA₂ from mitochondria decreases when mitochondria are incubated in the presence of respiratory substrates such as ADP, malate, and pyruvate, which causes an increase of transmembrane potential determined by cytofluorimetric analysis using DiOC₆(3) as a probe. The treatment of mitochondria with the uncoupler carbonyl cyanide 3-chlorophenylhydrazone slightly enhances sPLA₂ release. The increase of sPLA₂ specific activity after removal of mitochondrial outer membrane indicates that the enzyme is associated with mitoplasts. The mitochondrial localization of the enzyme has been confirmed by electron microscopy in U-251 astrocytoma cells and by confocal laser microscopy in the same cells and in PC-12 cells, where the structurally similar isoform type V-sPLA₂ has mainly nuclear localization. In addition to sPLA₂, mitochondria contain another phospholipase A₂ that is Ca²⁺-independent and sensitive to bromoenol lactone, associated with the outer mitochondrial membrane. We hypothesize that, under reduced respiratory rate, brain mitochondria release sPLA₂-IIA that might contribute to cell damage.

The energy metabolism of tumor cells relies on aerobic glycolysis rather than mitochondrial oxidation. This difference between normal and cancer cells provides a biochemical basis for new therapeutic strategies aimed to block the energy power plants of cells. The effects produced by the energy blockers bromopyruvate (3BP) and lonidamine (LND) and the underlying biochemical mechanisms were investigated in GL15 glioblastoma cells. 3BP exerts early effects compared to LND, even though both drugs lead cells to death but by different routes. A dramatic decrease of ATP levels occurred after 1 hour treatment with 3BP, followed by cytochrome c and hexokinase II degradation and by the decrease of both LC3I/LC3II ratio and p62, markers of an autophagic flux. In addition, Akt(Ser⁴⁷³) and p53(Ser¹⁵/Ser³¹⁵) dephosphorylation occurred. In LND treatment, sustained ATP cellular levels were maintained up to 40 hours. The autophagic response of cells was overcome by apoptosis that was preceded by phosphatidylinositol disappearance and pAkt decrease. This last event favored p53 translocation to mitochondria triggering a p53-dependent apoptotic route, as observed at 48 and 72 hours. Adversely, in 3BP treatment, phospho-p53 dephosphorylation targeted p53 to MDM2-dependent proteolysis, thus channeling cells to irreversible autophagy.