PSXV-13 The activity of thiol cathepsins in seminal plasma of stallions with different percentages of viable sperm after freezing-thawing

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 357-358
Author(s):  
Mikhail Atroshchenko ◽  
Anna Shitikova ◽  
Mariya Engalycheva
Keyword(s):  
New York ◽  
Seminal Plasma ◽  
Phase Contrast ◽  
Russian Science ◽  
Cathepsin H ◽  
Contrast Microscope ◽  
Normal Percentage ◽  
Viable Sperm ◽  
Oxidatively Modified ◽  
Low Activity

Abstract The aim of this work was to study the activity of cathepsins B, L and H in the seminal plasma of stallions with normal (>25%, n = 11) and low (< 25%, n = 13) percentage of viable sperm after freezing-thawing. Sperm of 24 Arabian stallions aged from 5 to 20 years (12.1 ± 4.8 years on average) was collected during the breeding season (February-May). The activity of cathepsins in spermoplasm was studied by the spectrofluorometric method (System 3 Scanning Spectrofluorometry, Optical technology devices, inc. Elmstord, New York, 10523) by Barrett&Kirschke. The viability of the sperm was determined after its freezing-thawing. Sperm smears were eosin stained and live:dead-ratio was examined using an Olympus BX41 phase contrast microscope (Olympus Corporation, Japan). The significance of differences in the studied groups was determined using the Mann-Whitney U-test. There were no significant differences in the activity of catepsins B and L in the spermoplasm of stallions with normal and low percentages of viable spermatozoa after freezing-thawing. It was found that in the group of stallions with a normal percentage of viable spermatozoa after freezing-thawing, the activity of cathepsin H in the spermoplasm was significantly higher than in the group with a low percentage of viable spermatozoa (P = 0.0219). Free radicals formed during freezing-thawing of sperm can damage cell membranes, leading to loss of sperm viability. Thiol cathepsins are involved in the degradation of oxidatively modified proteins and, apparently, it is cathepsin H that is most actively involved in this process in the sperm of stallions. We assume that the low activity of cathepsin H in the seminal plasma of stallions with a low percentage of viable spermatozoa indicates a high involvement of this enzyme in the protection of sperm membranes from oxidative damage. The research was supported by the Russian Science Foundation grant No. 20-16-00101.

Histochemical and Morphological Studies of Lipids in the Oogenesis of Pheretima posthuma

1958 ◽  
Vol s3-99 (48) ◽  
pp. 475-484
Author(s):  
VISHWA NATH ◽  
BRIJ L. GUPTA ◽  
S. L. MANOCHA
Keyword(s):  
Phase Contrast ◽  
Chemical Change ◽  
Follicular Epithelium ◽  
Cholesteryl Esters ◽  
Lipid Bodies ◽  
Interference Microscope ◽  
Phase Contrast Microscope ◽  
Morphological Studies ◽  
Contrast Microscope ◽  
Second Category

A study of the oocytes of the earthworm, Pheretima posthuma, examined fresh under the phase-contrast and interference microscopes as well as by histochemical techniques, has revealed that there are two types of lipid bodies in the cytoplasm. The lipid bodies of the first type (L1) are smaller, appear as homogeneous, dark granules under the phase-contrast microscope, and have a protein-phospholipid core surrounded by a thick sheath of phospholipids only. The lipid bodies of the second category (L2), which arise as a result of growth and chemical change in L1 bodies, have a pure phospholipid core surrounded by a thick triglyceride sheath. They give a ringed appearance under the phase-contrast microscope. The study under the interference microscope shows that this ringed appearance is an optical artifact. The lipid spheres present in the follicular epithelium contain phospholipids only. The mitochondria are in the form of minute granules. They remain unchanged throughout oogenesis. Some vacuoles devoid of any lipids, proteins, or carbohydrates have been observed. They also remain unchanged. Pure triglyceride spheres, yolk globules, nucleolar extrusions, as well as cholesterols and cholesteryl esters are absent.

Automated Confluence Measurement Method for Mesenchymal Stem Cell from Brightfield Microscopic Images

2021 ◽  
pp. 1-9
Author(s):  
Zenan Wang ◽  
Rucai Zhan ◽  
Ying Hu
Keyword(s):  
Cell Culture ◽  
Phase Contrast ◽  
Low Cost ◽  
Cost Effective ◽  
Cell Phenotype ◽  
Cell Culture Process ◽  
Reconstruction Method ◽  
Image Reconstruction Method ◽  
Contrast Microscope ◽  
Culture Process

Cell confluence is an important metric in cell culture, as proper timing is essential to maintain cell phenotype and culture quality. To estimate cell confluence, transparent cells are observed under a phase-contrast or differential interference contrast microscope by a biologist, whose estimations are error-prone and subjective. To overcome the necessity of using the phase-contrast microscope and reducing intra- and inter-observer errors, we have proposed an algorithm that automatically measures cell confluence by using a commonly used brightfield microscope. The proposed method consists of a transport-of-intensity equation-based brightfield microscopic image processing, an image reconstruction method, and an adaptive image segmentation method based on edge detection, entropy filtering, and range filtering. Experimental results have shown that our method has outperformed several popular algorithms, with an F-score of 0.84 ± 0.07, in images with various cell confluence values. The proposed algorithm is robust and accurate enough to perform confluence measurement with various lighting conditions under a low-cost brightfield microscope, making it simple and cost-effective to use for a fully automated cell culture process.

Redescription of Neelus fimbriatus Bretfeld & Trinklein, 2000 (Collembola: Neelidae) from Colombia

Zootaxa ◽  
2018 ◽  
Vol 4527 (3) ◽  
pp. 414
Author(s):  
ELIDA P. MARÍN ◽  
JOSÉ G. PALACIOS-VARGAS
Keyword(s):  
Phase Contrast ◽  
Phase Contrast Microscope ◽  
Contrast Microscope

Neelus fimbriatus is redescribed using specimens from Colombia. Drawings and phase contrast microscope photos of the species are used. New characters are used as tibiotarsal tuberculate setae and abdominal ventral acetabula. 

The Concentration of Salivary Steroid Hormones and the Prevalence of Gingivitis at Puberty

1988 ◽  
Vol 2 (2) ◽  
pp. 397-400 ◽  
Author(s):  
M. Morishita ◽  
H. Aoyama ◽  
K. Tokumoto ◽  
Y. Iwamoto
Keyword(s):  
Phase Contrast ◽  
Morphological Differentiation ◽  
Gingival Inflammation ◽  
Chi Square ◽  
Probing Depth ◽  
Whole Saliva ◽  
Contrast Microscope ◽  
Males And Females ◽  
Bacterial Plaque ◽  
Chi Square Analysis

Gingival conditions of 1323 junior high schoolchildren aged 12-15 were examined, and 132 children who had either healthy gingivae or severe gingivitis were called to the clinic. More precise examination of gingivitis was performed by assessment of Jackson's gingivitis index (G.I.), probing depth (P.D.), and bleeding on probing. Whole saliva was collected, and the salivary concentrations of estradiol, progesterone, and testosterone were determined by radioimmunoassay. Subgingival bacterial plaque was sampled from 36 children, and total bacterial counts and morphological differentiation were performed under a phase-contrast microscope. For statistical analysis, both males and females were divided into two groups according to the concentration of each sex hormone and subgrouped by the results of clinical examinations. Chi-square analysis using 2-by-2 tables was performed to determine the relation between salivary steroid hormone levels and gingival inflammation. The results suggest that unbalanced secretion of certain hormones might be one of the factors promoting gingivitis at puberty.

Methods of transplanting nuclei from single cultured cells to unfertilized frogs' eggs

Development ◽  
1970 ◽  
Vol 24 (2) ◽  
pp. 249-255
Author(s):  
J. B. Gurdon ◽  
R. A. Laskey
Keyword(s):  
Xenopus Laevis ◽  
Phase Contrast ◽  
Cultured Cells ◽  
The Other ◽  
Phase Contrast Microscope ◽  
Cell Method ◽  
Attached Cell ◽  
Contrast Microscope

Two methods of transplanting single nuclei from monolayers of cultured cells to unfertilized eggs of Xenopus laevis are described, illustrated, and tested. The detached-cell method is simpler and quicker to operate and is suitable for homogeneous populations of cells which are easily removed from the substrate on which they are growing. The other, attached-cell, method is technically more elaborate, but is applicable to cells whose properties can be individually determined under the phase-contrast microscope and to cells which are not readily dissociated from other cells or from their substrate.

scholarly journals Evaluation of Haematuria and Use of Phase Contrast Microscope: A Short Review

1970 ◽  
Vol 20 (1) ◽  
pp. 63-67 ◽  
Author(s):  
T Sultana ◽  
T Sultana ◽  
MQ Rahman ◽  
ANN Ahmed
Keyword(s):  
Phase Contrast ◽  
Diagnostic Marker ◽  
Short Review ◽  
Red Cells ◽  
Phase Contrast Microscopy ◽  
Red Cell ◽  
Phase Contrast Microscope ◽  
Non Invasive ◽  
Contrast Microscope ◽  
Contrast Microscopy

For centuries physicians have been using urine as one of the non-invasive means for assessing diseases. Haematuria is a frequently encountered abnormality in clinical practice. Haematuria may have either a glomerular or a non-glomerular origin. The morphological study of urinary red cells by Phase-Contrast Microscopy (PCM) is a useful diagnostic marker for glomerular bleeding, if correctly interpreted and used. Today, urinalysis and in particular identification of red cells morphology by Phase-Contrast Microscopy has been a widely accepted technique for determining the site of haematuria. A short review on haematuria and Phase-Contrast Microscopy are presented here for updating knowledge and academic interest. Key words: Phase-contrast microscope; Haematuria; Dysmorphic red cell. DOI: http://dx.doi.org/10.3329/jdmc.v20i1.8584 J Dhaka Med Coll. 2011; 20(1) :63-67

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