Infertility, the inability to conceive after one year of unprotected intercourse, affects about 15% of the global population. Nearly half of couples diagnosed with infertility have a contributing male or sperm factor (male-factor infertility). Preparation of purified sperm is an important component of fertility treatments. Most purification techniques rely on centrifugation, although the effects of centrifugation on sperm motility (movement) and DNA integrity are not well understood. Our objective is to manipulate centrifugation g-force and time settings following sperm preparation to investigate potential impacts of centrifugation on sperm quality for fertile and infertile men. Our hypothesis is that the highest centrifugation g-force and time will result in reduced motility and maximum DNA damage. Participants with normal semen parameters are recruited. A semen analysis is performed to determine volume, concentration, and baseline motility, which is measured using an automated visual sperm analyzer. Motile sperm are isolated via “direct swim out,” a standard purification technique. Nine different centrifugation settings are applied to the sample (time and g-force parameters studied: 7, 30, and 60 minutes, and 250xg, 600xg, and 800xg), and motility is evaluated. For each condition, DNA integrity is assessed with a COMET assay (uses single-cell gel electrophoresis to detect double-stranded DNA breaks). Preliminary results from 11 participants (ages 26-35) demonstrate substantial variability in motility for each participant. We observe that while the highest centrifugation g-force and time condition (60 minutes at 800xg) reduces motility for some participants, it has the opposite effect for others. Data collection for motility analysis and DNA integrity assays are in progress and will continue over the next three months. This project aims to provide high quality evidence to support current laboratory practices in sperm preparation, an essential part of fertility treatments.

Neural epidermal growth factor-like 1 (NELL1) is a protein that plays a significant role in bone and tissue development. Recent research has shown that NELL1 may also be involved in cancer progression, making it an important target for cancer research. In neuroendocrine prostate cancer (NEPC), the role of NELL1 remains largely unexplored, particularly in relation to the neuroendocrine features that make the cancer very aggressive. I investigated how NELL1 affects the transcription of principal genes associated with neuroendocrine differentiation, a process in which prostate cancer cells become similar to nerve and hormone-releasing cells, making the cancer more aggressive. Through quantitative Real-Time Polymerase Chain Reaction (qRT-PCR), I measured how the transcription of various genes changed when NELL1 levels were altered in prostate cancer cells. This technique quantifies mRNA levels, revealing genes' transcriptional activity. I found that changing NELL1 levels leads to changes in the transcription of certain genes that control neuroendocrine features of the cells. This suggests that NELL1 may crucially control prostate cancer aggressiveness. By understanding its relationship with key NEPC genes, this could lead to new treatment approaches and improved therapeutic drugs for patients.