Multiscale imaging of corneal endothelium damage and effects of Rho Kinase inhibitor application in mouse models of acute ocular hypertension.

We developed a multiscale optical imaging workflow, integrating and correlating visible-light optical coherence tomography, confocal laser scanning microscopy, and single-molecule localization microscopy to investigate the mouse cornea damages from the in-vivo tissue level to the nanoscopic single-molecule level. We used electron microscopy to validate the imaged nanoscopic structures. We imaged wild-type mice and mice with acute ocular hypertension and examined the effects of Rho Kinase inhibitor application. We defined four types of intercellular tight junction structures as healthy, compact, partially-distorted, and fully-distorted types by labeling the Zonula occludens-1 protein in the corneal endothelial cell layer. We correlated the statistics of the four types of tight junction structures with cornea thickness and intraocular pressure. We found that the population of fully-distorted tight junctions correlated well with the level of cornea edema, and applying Rho Kinase inhibitor reduced the population of fully-distorted tight junctions under acute ocular hypertension.

Ovarian function preservation in the cancer patient.

OBJECTIVE: To review the clinical modalities that are available to women receiving potentially sterilizing cancer therapy. DESIGN: The MEDLINE database was reviewed for all publications on medication, surgery, or assisted reproductive technology that could potentially preserve fertility in women who are receiving cancer therapy. CONCLUSION(S): There are many options available to a patient undergoing a treatment that will negatively impact her fertility. Many procedures and medical interventions have been proven successful both in terms of ovarian function and pregnancy rates. Other techniques have great potential but do not have long-term clinical data. It is important that the patient's primary care physician understand the methods available to preserve fertility in cancer patients and communicate this information to the patient.

The impact of peritoneal fluid from healthy women and from women with endometriosis on sperm DNA and its relationship to the sperm deformity index.

OBJECTIVE: To study the effect of peritoneal fluid (PF) from healthy women and from women with endometriosis on sperm DNA and its relationship to sperm morphology as assessed by the sperm deformity index (SDI). DESIGN: Experimental study. SETTING: Research laboratory at an academic hospital. PATIENT(S): Healthy women undergoing laparoscopic tubal ligation and women with endometriosis. INTERVENTION(S): Aliquots of prepared sperm from 10 healthy donors were incubated with PF from healthy women undergoing laparoscopic tubal ligation (treatment 1, n = 10), with PF from patients with endometriosis (treatment 2, n = 10), and with human tubal fluid media with 10% bovine serum albumin (control, n = 10). MAIN OUTCOME MEASURE(S): Sperm DNA fragmentation was assessed by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay and SDI. RESULT(S): There was a significant increase in sperm DNA damage at 24 hours compared with at 1.5 and 3 hours. The SDI scores significantly correlated with sperm DNA damage after 1.5 and 24 hours of incubation in all aliquots. Percentage normal morphology did not correlate with DNA damage. CONCLUSION(S): In vitro exposure of sperm to PF from patients with endometriosis is associated with significantly increased DNA damage. There is evidence of interdependence between the sperm morphology as assessed by SDI scores and DNA damage. The significant increase in sperm DNA damage observed after 24 hours of incubation may be clinically relevant.

Role of total antioxidant capacity in the differential growth of human embryos in vitro.

OBJECTIVE: The objective of this study was to examine the relationship of early human embryonic development parameters with day 1 culture media total antioxidant levels (day 1 TAC). DESIGN: Prospective study. SETTING: Patients undergoing assisted reproduction (ART) in a teaching hospital. PATIENT(S): Patients undergoing conventional IVF (n = 153; 167 cycles) and intracytoplasmic sperm injection (ICSI; n = 105; 116 cycles) were included. Both fertilization and early cultures were performed in human tubal fluid (HTF) with 5% serum substitute supplement. INTERVENTION(S): Levels of total antioxidants in the central well (sample) and the outer well (control) of each embryo culture dish were measured. MAIN OUTCOME MEASURE(S): Fertilization rate and embryo quality at days 3 and 5 were recorded for each cycle. Age, parity, and demographic features were also compared. RESULT(S): After controlling for all demographic and clinical variables, day 1 TAC was related to fertilization rates in both groups of patients. Day 1 TAC was significantly related to high (>7) day 3 cell number, low (<10%) fragmentation rate, and blastocyst development rate in both conventional IVF and ICSI cycles. Day 1 TAC was related to pregnancy in ICSI but not conventional IVF cycles. CONCLUSION(S): Day1 TAC appears to be an important biochemical marker for early embryonic growth. Decreased embryonic fragmentation, enhanced cleavage rate, and increased blastocyst development rate may be partially related to day 1 TAC in the culture media. Whether this relationship is a cause or effect needs further assessment.