Zofnat Wiener-Megnazi, et al FERTILITY AND STERILITY 2010

OBJECTIVE:
To examine whether the oxidative status of an individual embryo before transfer may predict chances of implantation.

DESIGN:
A prospective laboratory study.

SETTING:
An IVF unit in a university-affiliated hospital.

PATIENT(S):
One hundred thirty-three women undergoing IVF-ET treatment cycles.

INTERVENTION(S):
Before ET, 10 μL of embryo culture medium was retrieved individually from each embryo and the oxidative status assessed by the thermochemiluminescence (TCL) analyzer.

MAIN OUTCOME MEASURE(S):
The occurrence of pregnancy. Two parameters were recorded: the TCL amplitude after 50 seconds (H1) and the TCL ratio. These were compared with demographic, clinical, and laboratory parameters and treatment outcome. All data underwent statistical analysis.

RESULT(S):
Altogether 284 embryos were transferred in 133 ET cycles. Forty-one pregnancies occurred (31%). For embryos transferred after 72 hours (77 transfers), the highest H1 levels in each group of transferred embryos correlated with the occurrence of pregnancy. The combination of maximal intracohort H1 level <210 counts per second with a TCL ratio of ≤ 80% had a positive predictive value of 70.6% for the occurrence of pregnancy.

CONCLUSION(S):
The oxidative status of the early embryo in IVF is associated with the chances of implantation. Assessment of the oxidative status of embryos in culture media before transfer may serve as an applicable tool for improving embryo selection in light of the legal limitations of the number of transferred embryos allowed.

Link to PubMed

Zofnat Wiener-Megnazi, et al. FERTILITY AND STERILITY_ 82, SUPPL. 3, OCTOBER 2004

Abstract
OBJECTIVE:
To evaluate oxidative stress indices in follicular fluid (FF) by a novel thermochemiluminescence (TCL) assay and investigate the correlation between TCL and i.v.f. cycle parameters.

DESIGN:
Prospective, cross-sectional study.

SETTING:
I.v.f. Unit of an Obstetrics and Gynecology Department in a university-affiliated hospital.

PATIENT(S):
One hundred eighty-nine women undergoing consecutive i.v.f. treatment cycles during 2001.

INTERVENTION(S):
After oocyte retrieval, pooled FF was centrifuged and the supernatant was tested in the TCL assay.

MAIN OUTCOME MEASURE(S):
Maximal serum E2 levels, number of gonadotropin ampoules, retrieved oocytes, mature oocytes, fertilization and cleavage rates, number of available embryos and cryopreserved embryos, and clinical pregnancy rates and correlation with TCL indices.

RESULT(S):
The TCL curve slope of FF positively correlated with maximal serum E2 levels, number of mature oocytes, and number of cleaved embryos and was inversely correlated with the women’s ages and the number of gonadotropin ampoules. Follicular fluid TCL amplitude at 50 seconds ranged from 294 to 711 cps, but all pregnancies (n = 50; 28.1%) occurred within the range of 347-569 cps. With 385 and 569 cps as cutoff levels for the occurrence of pregnancy, the negative predictive value beyond this range was 96% and the positive predictive value within this range was 32%.

CONCLUSION(S):
The TCL results may reflect the age-related increase in free radical activity and is associated with parameters of ovarian responsiveness and IVF outcome. A certain threshold of oxidative stress may be required for the occurrence of conception in i.v.f. TCL is a potential tool to evaluate, treat, and monitor antioxidant therapy in i.v.f. treatments.

Link to PubMed

Lissak A¹, Wiener-Megnazi Z, Reznick AZ, Shnizer S, Ishai D, Grach B, Lahav-Baratz S, Shiloh H, Koifman M, Dirnfeld M.: FERTILITY AND STERILITY. 2004 Mar; 81 Suppl 1:792-7.

Abstract

OBJECTIVE:
To examine the effect of oxidation of proteins and lipids, as measured by a novel thermochemiluminescence (TCL) analyzer, and to evaluate the correlation between TCL indices in seminal plasma and sperm parameters.

DESIGN:
Experimental and prospective clinical studies.

SETTING:
An infertility unit.

PATIENT(S):
One hundred forty-eight men undergoing semen analysis.

INTERVENTION(S):
Bovine serum albumin (BSA) and linolenic acid were oxidized and tested by TCL, protein carbonyls, and conjugated dienes assays. All participants underwent semen analysis. Seminal plasma was tested by TCL and conjugated dienes.

MAIN OUTCOME MEASURE(S):
Thermochemiluminescence indices before and after oxidation of BSA and linolenic acid, compared with protein carbonyl and conjugated dienes indices. Correlation between semen parameters and TCL and conjugated dienes indices in seminal plasma.

RESULT(S):
Oxidation of BSA and linolenic acid was marked by characteristic changes in their TCL curve pattern and an increase in the levels of protein carbonyls and conjugated dienes. Among 125 sperm-containing semen samples, the TCL curve exhibited two patterns: a positive relative ratio curve (group A, 87 patients) and a negative relative ratio curve (group B, 38 patients). Sperm concentration was lower and total motile sperm and rapid motile sperm were fewer in group B. A significant correlation was found between TCL indices, conjugated dienes, and sperm quality in group B.

CONCLUSION(S):
Oxidation affects TCL curve pattern of proteins and lipids in a characteristic manner. Thermochemiluminescence indices in seminal plasma closely correlate with sperm characteristics among patients with sperm disturbances, and it might serve as a tool in the evaluation, treatment, and monitoring of subfertile men.

Link to PubMed

Shnizer S, Kagan T, Lanir A, Maor I, Reznick AZ: Luminescence 2003; 18:90–6.

ABSTRACT: Detection of electronically excited species (EES) in body fluids may constitute an important diagnostic tool in various pathologies. Examples of such products are triplet excited carbonyls (TEC), which can be a source for photon emission in the 400– 550 nm range. The aim of the present study was to determine the actual contribution of lipid and protein components (protein carbonyls) to photon emission generated by thermochemiluminescence (TCL) during the heating of biological fluids. In this study, a new TCL Photometer device, designed by Lumitest Ltd, Israel, was used. Samples were heated to a constant temperature of 80 _ 0.5C for 280 s and photon emission was measured at several time points. In order to compare the results of TCL measurements to conventional methods of detecting lipid and protein oxidation, each examined sample was also heated in a waterbath at 80C for 10–280 s. Lipid and protein oxidation were subsequently measured using conventional methods. The TCL of four polyunsaturated
fatty acids (PUFA) with three to six double bonds was measured. The elevation of the PUFA TCL amplitude correlated with the increase in the number of double bonds of PUFA. A correlation between the increase in TCL intensity and protein carbonyl generation in bovine serum albumin (BSA) was also observed. In the venous blood serum, our study showed that an increase of TCL intensity during heating reflected the cleavage of TEC of lipid origin. Our study suggests that biological molecules such as proteins, lipids and other molecules, which may become unstable during heating, are capable of generating EES. We demonstrated that a TCL curve can be used as a kinetic model for measuring oxidative processes, which reflects modifications of different molecules involved in the oxidative stress phenomena. Copyright _ 2003 John Wiley & Sons, Ltd.

Link to Pubmed

Download poster from here

Download poster from here