Biomedical subjects

Rakesh K Sharma

Publications and source records attributed to Rakesh K Sharma.

58 records · Page 4Linked to original sources

PubMedDate not supplied

The use of novel semen quality scores to predict pregnancy in couples with male-factor infertility undergoing intrauterine insemination.

The purpose of this study was to determine whether 2 new semen quality (SQ) scores could predict pregnancy in patients undergoing intrauterine insemination (IUI) for male-factor infertility and whether an overall score could help in counseling these couples with assisted reproductive technique (ART) options. Ninety-three couples with male-factor infertility were examined for semen analysis. Samples were prepared by density gradient separation (47% and 90%), and IUI was performed. On the basis of the 2 semen scores (SQ and relative quality [RQ]), the IUI-semen pregnancy score (IUI-SPS) was calculated. Of the 192 IUI cycles, 14% (27 of 192) resulted in pregnancy. Both prewash SQ and RQ scores were significantly related to pregnancy (P =.02 and P <.001), as was the postwash RQ score (P <.001). Of the IUI cycles in which the postwash RQ score was greater than 125, 40% (13 of 32) resulted in pregnancy compared to 9% of cycles (14 of 160) in which the postwash SQ score was less than 125. The prewash IUI-SPS score was significantly related to IUI-induced pregnancy (P <.001). Both the pre- and postwash SQ and RQ scores can predict pregnancy in male infertility patients undergoing IUI. Patients with an IUI-SPS less than 150 may be advised to seek in vitro fertilization (IVF), whereas those with an IUI-SPS greater than 150 may be advised to seek IUI.

Female↗

PubMedDate not supplied

Enhanced chemiluminescence assay vs colorimetric assay for measurement of the total antioxidant capacity of human seminal plasma.

Although the enhanced chemiluminescence assay is commonly used to measure the nonenzymatic total antioxidant capacity (TAC) of the human seminal plasma, it is cumbersome, expensive, and time-consuming. We describe herein an alternate method to measure TAC that is based on the ability of antioxidants in seminal plasma to interfere with a reaction between 2,2'-azino-di-[3-ethylbenzthiazoline sulphonate] and metmyoglobin with H(2)O(2). This reaction produces a relatively stable blue-green color with absorbance maxima at 600 nm. We compared this colorimetric assay with our established chemiluminescence method and assessed quality control parameters (ie, intra-assay and interassay variabilities) in addition to intraobserver and interobserver differences. Our results show that the colorimetric assay was fairly predictive of antioxidant capacity similar to the chemiluminescence assay (P <.001). Furthermore, there was a high level of agreement between the duplicate measures by the same observer (intraobserver) and intra-assay variability, with a concordance correlation coefficient of 0.99. The interassay coefficient of variation was 4.7% (overall). The mean +/- SD of the difference between the 2 observers was 2.98% +/- 4.1%. In conclusion, we found that the colorimetric assay is a reliable and accurate method to evaluate seminal TAC, and it could be used as a simpler, rapid, and cheaper alternative to the chemiluminescence assay.

Antioxidants↗

PubMedDate not supplied

Utility of the nitroblue tetrazolium reduction test for assessment of reactive oxygen species production by seminal leukocytes and spermatozoa.

The purpose of this study was to evaluate the ability of spermatozoa and leukocytes in semen to produce reactive oxygen species (ROS) by using nitroblue tetrazolium (NBT) staining and to examine the association between NBT staining and levels of ROS as measured by chemiluminescence. Twenty-one infertility patients (leukocytospermia; n = 8; nonleukocytospermia, n = 13) and 9 healthy donors were included. Standard semen analysis and density gradient centrifugation were performed to test NBT staining, ROS, and total antioxidant capacity. A ROS-total antioxidant capacity (ROS-TAC) score was calculated by using principal component analysis. In the leukocytospermic group, after separation on a density gradient, the percentage of NBT-positive staining was significantly higher in sperm suspensions contaminated with leukocytes (median [25th, 75th percentiles]; 70% [61%, 79%]) compared to the nonleukocytospermic group (14.5% [9%, 25.5%]; P =.03) and donors (7% [3%, 11%]; P =.02), respectively. A strong positive correlation was seen between levels of ROS in whole ejaculates and NBT-positive staining in leukocytes (r = 0.59; P <.0006) and in leukocyte fractions (r = 0.72; P <.0001) after density gradient separation. Similarly, ROS was positively correlated with excessive cytoplasmic retention in spermatozoa from whole ejaculates and abnormal spermatozoa after separation on density gradients (r = 0.72; P <.0001). The ROS-TAC score was inversely correlated with NBT staining in leukocytes in whole ejaculates (r = -0.960, P <.0007) and in both leukocyte fractions (r = -0.39; P <.04) and spermatozoa with cytoplasmic retention (r = -0.38; P <.04). Our results indicate that the NBT reduction test can be used to assess the contribution of seminal leukocytes and defective spermatozoa towards ROS generation in semen. Levels of ROS assessed by chemiluminescence assay are strongly correlated with the results of NBT staining.

Antioxidants↗

PubMedDate not supplied

Effects of temperature on sperm motion characteristics and reactive oxygen species.

OBJECTIVE: To evaluate the effect of temperature on sperm motility, sperm motion characteristics, and production of reactive oxygen species (ROS) in semen. METHODS: We collected semen samples from 12 infertile patients and 12 healthy donors and divided each sample into four aliquots just after liquefaction. One aliquot was analyzed immediately after liquefaction and the remaining three were analyzed after one hour of incubation at 4 degrees C, 25 degrees C, and 37 degrees C. Sperm motility and motion characteristics were measured by computer-assisted sperm analysis (CASA). Levels of ROS were determined in washed sperm suspensions using a chemiluminescence assay and results were expressed as x10(6) counted photons per minute (cpm)/20 million sperm/mL. RESULTS: Motility, curvilinear velocity, straight-line velocity, average path velocity, and amplitude of lateral head displacement decreased significantly from baseline values in both the patients and donors after incubation at 4 degrees C. Percentage motility was highest and ROS levels were lowest in the samples that were incubated at 37 degrees C. CONCLUSION: Based on our results, we suggest that semen samples be stored at 37 degrees C after collection and during transportation and processing.

Adult↗