PSII-B-24 Use of the Elnady technique of tissue preservation to preserve specimens for use in undergraduate animal science courses

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 503-503
Author(s):  
Jay A Daniel ◽  
Isabella Kukor
Keyword(s):  
Reproductive Tract ◽  
Female Reproductive Tract ◽  
Tissue Preservation ◽  
Animal Science ◽  
Fresh Tissue ◽  
Tissue Samples ◽  
Science Courses ◽  
Undergraduate Instruction ◽  
Plastic Bag ◽  
Tissue Specimens

Abstract The use of tissue specimens for undergraduate instruction is a very valuable tool. However, fresh tissue specimens are not always available and many common preservation techniques can result in discoloration, offensive odors, and/or dangerous chemical residues. The Elnady Technique was developed as a means to produce tissue specimens that “are realistic, durable, have no offensive odor, and are dry, soft and flexible” (Elnady, F.A. 2016 The Elnady Technique: An innovative, new method for tissue preservation. Altex. 33:237–242. doi:10.14573/altex.1511091). Briefly for soft tissue, specimens were preserved by fixing in formalin for one week. The tissue specimen was then dehydrated with a series of acetone baths changed weekly. Once the tissue was fully dehydrated, the specimen was impregnated in glycerin by full immersion in a glycerin bath for one to two weeks. Excess glycerin was then removed by draining followed by immersion in cornstarch for one to two weeks. Cornstarch residue was removed with a soft brush, and the specimen was stored in a plastic bag. Multiple specimens (including female reproductive tract of the cat, goat, horse, and sow; digestive tract of barred owl, cat, chicken, and dog; one day old lamb stomach; goat rumen, reticulum, omasum, and abomasum; deer testicle; and sheep heart and kidney) have been preserved and used in various animal science course laboratories (75 laboratory sections and over 1000 students at Berry College). Some of the specimens have been in use for five years and are still in usable condition. The Elnady Technique has proven to be a useful means of preserving tissue samples used in undergraduate animal science courses.

Granulocyte - macrophage colony stimulating factor and interleukin-8 in the reproductive tract of ewes following oestrus and mating

2007 ◽  
Vol 19 (4) ◽  
pp. 585 ◽  
Author(s):  
Jennifer L. Scott ◽  
Natkunam Ketheesan ◽  
Phillip M. Summers
Keyword(s):  
Reproductive Tract ◽  
Staining Intensity ◽  
Female Reproductive Tract ◽  
Enzyme Linked Immunosorbent Assay ◽  
Colony Stimulating Factor ◽  
Tissue Samples ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

Cytokines produced in the female reproductive tract after mating may enhance reproductive success. The present study investigated the distribution of granulocyte–macrophage colony stimulating factor (GM-CSF) and interleukin (IL)-8 in tissues and luminal secretions from different sites in the reproductive tract of the ewe following oestrus and after natural mating. Fifteen ewes were mated with a ram for 1 h and their reproductive tracts collected 3, 6, 18, 24 or 48 h later. Another 15 ewes were used as oestrous controls. Luminal secretions and tissue samples were collected from seven sites in each reproductive tract. Secretions were analysed by enzyme-linked immunosorbent assay and tissues were stained immunohistochemically using anti-sheep GM-CSF and anti-sheep IL-8 antibodies. Both cytokines were found in luminal and glandular endometrial epithelium and, to a lesser extent, in cervical epithelium; neither was found in the vaginal epithelium. Twice as many (P < 0.05) luminal samples from mated ewes than non-mated ewes were positive for GM-CSF. The vaginal lumen contained significantly higher (P < 0.01) concentrations of IL-8 compared with other sites, irrespective of mating status. Significant differences (P < 0.05) were found in staining intensity of GM-CSF and IL-8 from different sites. Production of GM-CSF and IL-8 by reproductive tissues is likely to contribute to leucocyte infiltration into the ovine reproductive tract.

288. The post-insemination inflammatory response in the ewe

2005 ◽  
Vol 17 (9) ◽  
pp. 121 ◽  
Author(s):  
J. L. Scott ◽  
N. Ketheesan ◽  
P. M. Summers
Keyword(s):  
Inflammatory Response ◽  
Reproductive Tract ◽  
Female Reproductive Tract ◽  
Enzyme Linked Immunosorbent Assay ◽  
Tissue Samples ◽  
Gm Csf ◽  
Reproductive Tissues ◽  
Cell Counts ◽  
Macrophage Colony ◽  
Uterine Body

Insemination causes an inflammatory response in the female reproductive tract of many species. The cytokine/leukocyte network initiated during this reaction is believed to enhance reproductive success.1 This study investigated the post-insemination inflammatory response in the ewe. Fifteen nonparous ewes were mated with the same ram for 1 h and their reproductive tracts were collected 3, 6, 18, 24 or 48 h later. Another fifteen ewes were used as controls. Tissue samples and luminal mucus were collected from 10 sites in each reproductive tract and stained with haematoxylin and eosin, Diffquik and immunohistochemically using a monoclonal CD68 antibody to quantify neutrophils, eosinophils and macrophages. Presence of interleukin-8 (IL-8) and granulocyte-macrophage colony-stimulating factor (GM-CSF) was investigated using immunohistochemistry and enzyme-linked immunosorbent assay. Neutrophils and macrophages increased in reproductive tissues following insemination. Mean cell counts in 1.5-mm2 tissue of mated (M) and control (C) ewes demonstrated a peak in neutrophils at 6–18 h post-insemination with significant differences (P < 0.05) between mated and controls in the posterior cervix (M = 23.7; C = 4.1) and uterine body (M = 34.5; C = 11.5). Macrophages peaked at 18–24 h with significant differences (P < 0.05) between mated and controls in the vagina (M=13.4; C = 4.6), posterior cervix (M = 10.4; C = 2.7), mid-cervix (M = 8.5; C = 3.0) and ipsilateral mid-uterine horn (M = 14.2; C = 7.9). Neutrophils increased in the lumen of the cervix and uterine body following insemination but macrophage numbers did not change. Insemination did not affect eosinophils. IL-8 and GM-CSF were detected in endometrial epithelial cells in mated and non-mated ewes. Highest concentrations of IL-8 were found in vaginal mucus. Small quantities of GM-CSF were detected in occasional mucus samples. No difference between mated and non-mated ewes was demonstrated for either cytokine. In conclusion, the post-insemination inflammatory reaction in the ewe involves an increase in neutrophils and macrophages in reproductive tissues, with neutrophils crossing the epithelium into the lumen. There was no apparent increase in IL-8 or GM-CSF in response to insemination. (1)Robertson SA et al. (1997) American Journal of Reproductive Immunology 37, 438–442.

Evaluation of intestinal cancer by 1H HR-MAS NMR spectroscopy: A new diagnostic tool?

2021 ◽  
Vol 6 (3) ◽  
pp. 175-181
Author(s):  
Faraz Ahmad ◽  
Manish Agrawal ◽  
V K Singh ◽  
Mukesh Kumar ◽  
Surendra Kumar ◽  
...  
Keyword(s):  
Malignant Tumors ◽  
Ex Vivo ◽  
Mas Nmr ◽  
Fresh Tissue ◽  
Tissue Samples ◽  
Tumor Margin ◽  
Metabolic Characteristics ◽  
Significant Difference ◽  
Tissue Specimens ◽  
Sample Heterogeneity

 Previous studies have demonstrated that cancer cells harbor unique metabolic characteristics relative to healthy counterparts. The current study is a prospective ex-vivo HR-MAS NMR analysis of malignant colorectal cancer (CRC) tissue specimens and its corresponding benign tissues.To assess the HR-MAS Spectroscope qualitatively & to analyze significant difference between the normal, benign and malignant intestinal mucosa.Between November 2013and January 2016, 36 consecutive patients with confirmed CRC were recruited to a prospective observational study. Fresh tissue samples were obtained from center of tumor and 5 cm from tumor margin from surgical resection specimens. Samples were run in duplicate where tissue volume permitted to compensate for anticipated sample heterogeneity. Typically, the sample was packed into a 4 mm ZrO2 rotor of 50 μl capacity; a volume of 20μl of D2O having 0.03% TSP was used as a chemical shift reference. The sample-rotor-setup was then transferred into the HR-MAS NMR probe for analysis.A total of 36 spectra were acquired (center of tumor, n = 18; 5 cm from tumor margin, n = 18). The malignant clustering occurs due to increased Val (0.90ppm), Lac (1.34ppm), Ala(1.48ppm) levels of acetate (1.90ppm), glutamate (2.35ppm), taurine (3.23 ppm), choline containing compounds (3.20-3.22ppm), glycine (3.56ppm), lactate (4.12ppm) and α-H of Leu, Ileu, Val, Lys, Ala (3.76-3.79ppm . In addition unique metabolic profiles were observed for tumors of differing T-stage. The information gathered from clustering in PCA had highly suggested that malignancy induces metabolic perturbations at cellular levels.HR-MAS NMR profiling demonstrates cancer-specific metabolic signatures in CRC and reveals metabolic differences between benign and malignant tumors. In addition, this approach reveals that tumor metabolism undergoes modification during local tumor advancement, offering potential in future staging and therapeutic approaches.

scholarly journals A Comparison of Six Monoclonal Antibodies for Detection of Cytokeratins in Normal and Neoplastic Canine Tissues

1989 ◽  
Vol 1 (4) ◽  
pp. 316-323 ◽  
Author(s):  
Aglaia Cardona ◽  
Bruce Raymond Madewell ◽  
Diane Kathy Naydan ◽  
Judi Kay Lund
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibodies ◽  
Immunoperoxidase Method ◽  
Human Tissues ◽  
Fresh Tissue ◽  
Tissue Samples ◽  
Intermediate Filament Proteins ◽  
Epithelial Tissues ◽  
Epithelial Neoplasms ◽  
Tissue Specimens

Twenty normal canine tissue specimens, both fetal and adult; 19 epithelial neoplasms; and 18 nonepithelial neoplasms were examined using 6 commercially available monoclonal antibodies differing in their recognition of various molecular weight cytokeratins in human tissues. Fresh tissue samples were fixed in 100% ethanol and paraffin embedded prior to sectioning. The intermediate filament proteins were identified by an avidin-biotin-immunoperoxidase method. Primary antisera used included AE1/AE3, CAM-5.2, 35BH11, 34BE12, PKK1, MAK-6 cytokeratins, and vimentin. Monoclonal antibodies detected cytokeratins in a wide variety of canine epithelial tissues and neoplasms. Normal mesenchymal tissues and neoplasms, and stromal elements of epithelial tissues, showed no reactivity with anti-cytokeratins, but reacted positively with vimentin. Although PKK1, CAM-5.2, and MAK-6 were the most consistently reactive anti-cytokeratins, the full panel of monoclonals was required to detect cytokeratins in all of the epithelia evaluated.

THE UPTAKE OF [6,7-3H]OESTRADIOL BY THE NORMAL HUMAN FEMALE REPRODUCTIVE TRACT

1967 ◽  
Vol 39 (4) ◽  
pp. 599-607 ◽  
Author(s):  
M. G. BRUSH ◽  
R. W. TAYLOR ◽  
R. J. B. KING
Keyword(s):  
Menstrual Cycle ◽  
Luteal Phase ◽  
Reproductive Tract ◽  
Specific Activity ◽  
Follicular Phase ◽  
Female Reproductive Tract ◽  
Cell Fractionation ◽  
Tissue Samples ◽  
Essentially Normal ◽  
Radioactivity Content

SUMMARY The uptake of [6,7-3H]oestradiol by the reproductive tract was studied in 18 non-pregnant women who were undergoing hysterectomy for conditions not associated with an abnormal endometrium. A single s.c. injection of 50 μc [6,7-3H]oestradiol (specific activity, 182 μc/μg.) was given 2 hr. before hysterectomy. Tissue samples were dissolved in 5 n-alkali and their radioactivity content determined by liquid scintillation counting. Total and free steroid (ether soluble) radioactivities in the plasma were also determined. The existence of an essentially normal menstrual cycle in these patients was confirmed by histological examination of tissue samples and, in some cases, by determinations of urinary pregnanediol. Urinary total radioactivity was within the normal range, except in one case, thus confirming normal oestrogen metabolism. The radioactivity content of the tissues of the reproductive tract was well above that attributable to their plasma content. In some cases the uptake by different regions of the endometrium varied considerably. In the follicular phase of the menstrual cycle uptake was higher than that in the luteal phase. Vaginal and cervical epithelium showed variable uptake but in general it was lower than that for endometrium. Cell fractionation studies showed that, in general, proportionally more of the endometrial radioactivity was associated with the nuclei in the follicular phase than in the luteal phase.

Effect of Contraceptive Steroids on the Endometrium of Guinea Pig: SEM study

1977 ◽  
Vol 35 ◽  
pp. 668-669
Author(s):  
Mai M. Said ◽  
Ramesh K. Nayak ◽  
Randall E. McCoy
Keyword(s):  
Guinea Pig ◽  
Reproductive Tract ◽  
Three Dimensional ◽  
Female Reproductive Tract ◽  
Human Female ◽  
Surface Ultrastructure ◽  
Transmission Electron ◽  
Sem Study ◽  
Uterine Mucosa ◽  
Group 1

Burgos and Wislocki described changes in the mucosa of the guinea pig uterus, cervix and vagina during the estrous cycle investigated by transmission electron microscopy. More recently, Moghissi and Reame reported the effects of progestational agents on the human female reproductive tract. They found drooping and shortening of cilia in norgestrel and norethindrone- treated endometria. To the best of our knowledge, no studies concerning the effects of mestranol and norethindrone given concurrently on the three-dimensional surface features on the uterine mucosa of the guinea pig have been reported. The purpose of this study was to determine the effect of mestranol and norethindrone on surface ultrastructure of guinea pig uterus by SEM.Seventy eight animals were used in this study. They were allocated into two groups. Group 1 (20 animals) was injected intramuscularly 0.1 ml vegetable oil and served as controls.

Surface ultrastructure of danazol treated rat endometrium: A SEM study

1983 ◽  
Vol 41 ◽  
pp. 556-557
Author(s):  
R.P. Apkarian ◽  
J.S. Sanfilippo
Keyword(s):  
Cross Sections ◽  
Reproductive Tract ◽  
Preliminary Investigation ◽  
Breast Disease ◽  
Distal Segment ◽  
Uterine Horn ◽  
Female Reproductive Tract ◽  
Ultrastructural Changes ◽  
Cycle Phase ◽  
Sprague Dawley

The synthetic androgen danazol, is an isoxazol derivative of ethisterone. It is utilized in the treatment of endometriosis, fibrocystic breast disease, and has a potential use as a contraceptive. A study was designed to evaluate the ultrastructural changes associated with danazol therapy in a rat model. The preliminary investigation of the distal segment of the rat uterine horn was undertaken as part of a larger study intended to elucidate the effects of danazol on the female reproductive tract.Cross-sections (2-3 mm in length) of the distal segment of the uterine horn from sixteen Sprague-Dawley rats were prepared for SEM. Ten rats in estrus served as controls and six danazol treated rats were noted to have alterations of the estrus cycle i.e. a lag in cycle phase or noncycling patterns. Specimens were fixed in 3% glutaraldehyde in 0.05M phosphate buffer containing CaCl2 at pH 7.0-7.4 and chilled to 4°C. After a brief wash in distilled water, specimens were passed through a graded series of ethanol, critical point dryed in CO2 from absolute ethanol, and coated with 6nm Au. Observations were made with an IS1-40 SEM operated at 15kV.

The application of electron microscopy to diagnosis in gynecologic neoplasms and tumor-like conditions

1984 ◽  
Vol 42 ◽  
pp. 102-105
Author(s):  
Lawrence M. Roth
Keyword(s):  
Electron Microscopy ◽  
Reproductive Tract ◽  
Malignant Tumors ◽  
Frozen Section ◽  
Cost Effective ◽  
Female Reproductive Tract ◽  
Ultrastructural Examination ◽  
Specific Diagnosis ◽  
Potential Value ◽  
Gynecologic Neoplasms

The female reproductive tract may be the site of a wide variety of benign and malignant tumors, as well as non-neoplastic tumor-like conditions, most of which can be diagnosed by light microscopic examination including special stains and more recently immunoperoxidase techniques. Nevertheless there are situations where ultrastructural examination can contribute substantially to an accurate and specific diagnosis. It is my opinion that electron microscopy can be of greatest benefit and is most cost effective when applied in conjunction with other methodologies. Thus, I have developed an approach which has proved useful for me and may have benefit for others. In cases where it is deemed of potential value, glutaraldehyde-fixed material is obtained at the time of frozen section or otherwise at operation. Coordination with the gynecologic oncologist is required in the latter situation. This material is processed and blocked and is available if a future need arises.

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