Sheng Jing Decoction Can Promote Spermatogenesis and Increase Sperm Motility of the Oligozoospermia Mouse Model

Sheng Jing Decoction (SJD), as a traditional Chinese medicine prescription, is mainly be used to treat male infertility. However, the pharmacological functions and molecular mechanisms of SJD are poorly understood. In this study, we investigated the functions of SJD on spermatogenesis and sperm motility and explored the potential mechanisms involved. Here, we demonstrated that high, medium, and low doses of SJD are effective in restoring the impairments of the whole body and testicular tissue by cyclophosphamide inducing and to rescue the damage of testicular tissue cells including Sertoli cells and germ cells. SJD can partly restore the decrease in sperm concentration, sperm vitality, sperm motility, and normal sperm morphology rate in oligozoospermic mouse models. Ki67 staining analyses confirm SJD can promote testicular tissue cell proliferation. Real-time RT-PCR analyses also reveal that SJD can upregulate the expression of proliferation-associated gene Lin28a and differentiation-associated genes Kit, Sohlh2, and Stra8. SJD can also reduce the impairment of mitochondrial membrane potential (MMP) and sperm plasma membrane integrity by cyclophosphamide inducing. Our results reveal that SJD is effective in improving both sperm quantity and quality by increasing the sperm concentration, sperm vitality, sperm motility, and normal sperm morphology rate. SJD can promote spermatogenesis by upregulating the expression of the proliferation-associated gene Lin28a and the differentiation-associated genes (Kit, Sohlh2, and Stra8). SJD can sustain MMP and sperm plasma membrane integrity to increase sperm motility.

Polystyrene nanoplastics and microplastics can act as Trojan horse carriers of benzo(a)pyrene to mussel hemocytes in vitro

In this work we studied the ability of polystyrene (PS) nanoplastics (NPs) and microplastics (MPs) to transfer benzo(a)pyrene (BaP) to mussel hemocytes and to produce toxic effects in vitro. For this, intracellular fate and toxicity of PS NPs (0.05 μm) and MPs (0.5 and 4.5 μm) alone or with BaP and of BaP alone were assessed. Particles of 0.05 and 0.5 µm largely aggregated in the exposure medium whereas presence of BaP reduced particle aggregation. Cells internalized PS NPs and MPs alone or with BaP and these were found inside and outside lysosomes, depending on their size. PS particles alone or with BaP were cytotoxic to hemocytes only at the highest concentrations tested. The same was true for most sublethal endpoints except for increased phagocytic activity provoked by NPs and 0.5 μm MPs at lower concentrations. Plastic particles appeared to be the main drivers for reduced plasma membrane integrity and increased phagocytic and lysosomal activities whereas BaP appeared to contribute more to reduced cell viability and phagocytosis and increased ROS production and genotoxicity. Overall, PS NPs and MPs can act as carriers of BaP to mussel hemocytes, rising concerns about risks plastics associated to pollutants may pose to aquatic organisms.

Lipidomic analysis of tissue culture cells, tissues, and purified organelles v1

Lipids are a class of molecules that have roles in energy storage, plasma membrane integrity, and signaling events. To gain more understanding of the functions and roles that lipids play in biology, researchers employ discovery analytical approaches, such as mass spectrometry (MS)-based lipidomics. The main objective of this protocol is to provide directive on how to extract lipids from plasma, cells, tissue, and purified organelles for analysis by liquid chromatography (LC)-MS. This analysis will typically yield quantitative data for more than 200 lipids, depending on the sample type analyzed, across a range of lipid classes: phospholipids, cardiolipins, sphingolipids, di- and triacylglyerols, and cholesteryl-esters.

In vitro effect of codeine on human sperm motility and DNA integrity

Purpose This study assessed the in vitro effect of codeine, a popular drug of abuse, on human spermatozoa motility, plasma membrane integrity, DNA integrity, and oxidative stress. Materials and Methods Semen samples were collected from fifteen healthy donors and conventional semen analysis was carried out per the guideline of the World Health Organization. Direct Swim-up technique was performed to obtain highly motile sperm. Samples were incubated at 34.5°C with different concentrations (0, 0.1, 1, 5 and 10 mM) of codeine. The non-exposed (0 mM) was used as the control group. Sperm motility and DNA integrity were assessed at 30, 60, and 90 minutes, while sperm membrane integrity and sperm 8-OHdG level were determined at 90 minutes. Results Codeine at any tested concentration significantly reduced sperm motility and plasma membrane integrity but increased sperm 8-OHdG level compared to the control in a time-dependent manner. Furthermore, codeine at 1, 5, and 10 mM markedly increased sperm DNA damage. In addition, correlation study showed that sperm 8OHdG level was negatively associated with sperm motility, plasma membrane integrity, and DNA integrity. Conclusions Codeine may impair human spermatozoa fertilization capacity by inducing sperm dysmotility and damage to the sperm plasma membrane and DNA through an oxidative stress-dependent mechanism.

Assessment of tebuconazole exposure on bovine testicular cells and epididymal spermatozoa

This study is the first to investigate the effects of tebuconazole (TEB) on the physiological functions of bovine testicular cells and epididymal spermatozoa. Motility and plasma membrane integrity of spermatozoa exposed to TEB (0.001–100 µM) were evaluated at different incubation times (0–6 h), while TEB-induced spermiotoxicity was assessed after 24 h in cell cultures. Testicular cells, obtained from the parenchyma of bovine testes, were seeded at 1.0 × 104 and 1.5 × 106 cells/well in 96- and 12-well culture plates and incubated for 48 h in culture media containing TEB (0.001–100 µM) to evaluate cytotoxicity and hormone release, respectively. TEB did not affect the motility and plasma membrane integrity. However, significant spermiotoxicity occurred at higher TEB (1–100 µM) concentrations (P < 0.05) compared to control and lower doses. Although no dose caused cytotoxicity in testicular cells (P > 0.05), 1 and 100 µM TEB caused a significant increase in testosterone secretion (P < 0.05). As a result, high doses of TEB (1–100 µM) had slightly suppressive effects on spermatozoa; however, these doses had stimulatory effects on testosterone secretion by testicular cells. It appears that the disruption of hormonal homeostasis of testicular cells after TEB exposure may result in metabolic and especially reproductive adverse effects in bulls.

α-Fodrin in Cytoskeletal Organization and the Activity of Certain Key Microtubule Kinesins

Cortical cytoskeletal proteins are significant in controlling various cellular mechanisms such as migration, cell adhesion, intercellular attachment, cellular signaling, exo- and endocytosis and plasma membrane integrity, stability and flexibility. Our earlier studies involving in vitro and ex vivo approaches led us to identify certain undiscovered characteristics of α-fodrin, a prominent cortical protein. The conventional functions attributed to this protein mainly support the plasma membrane. In the present study, we utilized a global protein expression analysis approach to detect underexplored functions of this protein. We report that downregulation of α-fodrin in glioblastoma cells, U-251 MG, results in upregulation of genes affecting the regulation of the cytoskeleton, cell cycle and apoptosis. Interestingly, certain key microtubule kinesins such as KIF23, KIF2B and KIF3C are downregulated upon α-fodrin depletion, as validated by real-time PCR studies.

Response of Lawn Grasses to Salinity Stress and Protective Potassium Effect

The salinity effects on lawn grasses caused by mine salts (halite and carnallitite) due to road de-icing processes was the aim of this study. Biometric and physiological parameters were evaluated after salt dosage of 50 and 100 g m−2 applied to a lawn surface twice and four times, in weekly intervals. The alleviating effect to the salinity on the grasses from potassium enriched soil was also evaluated. Protective effect of potassium included mostly plasma membrane integrity and an increase in the level of photosynthetic pigments. This probably resulted in more efficient photosynthesis and thus increased lawn growth. Simultaneously, only a slight reduction in relative water content (RWC) was noted, so the recorded increase in proline level may indicate its participation in osmotic adjustment. Our results confirm the importance of proper, and even over-optimal, potassium fertilization of lawn grasses exposed to salinity. Moreover, it is advisable to use other fossil salts instead of halite for the de-icing of near-green areas. The mined salt carnallitite which, besides NaCl, contains about 30% of carnalite (KCl·MgCl2·6H2O) could be such a substance.

Plasma membrane integrity: implications for health and disease

Plasma membrane integrity is essential for cellular homeostasis. In vivo, cells experience plasma membrane damage from a multitude of stressors in the extra- and intra-cellular environment. To avoid lethal consequences, cells are equipped with repair pathways to restore membrane integrity. Here, we assess plasma membrane damage and repair from a whole-body perspective. We highlight the role of tissue-specific stressors in health and disease and examine membrane repair pathways across diverse cell types. Furthermore, we outline the impact of genetic and environmental factors on plasma membrane integrity and how these contribute to disease pathogenesis in different tissues.

Epididymal tail solid-surface vitrification as an effective method for domestic cat sperm cryobanking

Summary This study aimed to describe the viability of domestic feline spermatozoa after epididymal tail vitrification. For this, 10 pairs of testis–epididymis complexes were used. The epididymal tails were vitrified using the solid-surface vitrification (SSV) method, in which two vitrification media containing ethylene glycol (EG) 40% or glycerol (GLY) 40% were tested. Vitrification with the presence of EG resulted in better results for all sperm motility parameters (motility, vigour and CASA) compared with GLY (P < 0.05). There were no statistical differences for sperm viability and acrosome integrity, plasma membrane integrity, or overall health of morphologically normal sperm before or after vitrification among experimental groups. In conclusion, epididymal tail vitrification appears to be a suitable method for long-term storage of cat sperm, especially if the procedure is performed with EG as the cryoprotectant.