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Animal Reproduction 2021is endemic species in Indonesia. Study about fetal development of are very rare because of sample limitation. This study was carried out to describe the morphometrics...
is endemic species in Indonesia. Study about fetal development of are very rare because of sample limitation. This study was carried out to describe the morphometrics and x-ray analysis of three fetuses in different stage to give basic information about fetal development of . Three fetus samples fixed in Bouin's solution was used in this study. Observation was carried out to identify the characteristic of three fetus samples. This included the pattern of hair, body measurements, body volume, and body weight. X-ray analysis was carried out to know the ossification process in the fetal development. Statistical analysis was carried out using Microsoft 365® Excel program software. Three fetus samples had different specific hair pattern, that was hairless, smooth hairs, and smooth hairs with dense-non dense pattern. Body volume of 1, 2, and 3 fetus were 23mL, 90mL, and 170mL, respectively. Body weight of 1, 2, and 3 fetus were 19.5g, 79.22g, and 153.18g, respectively. Pearson's correlation analysis shown strong relationship between total body length, front body length, back body length, horizontal body diameter, vertical body diameter, head length, and head diameter against body volume and body weight of three fetuses. Significant positive correlation was shown between horizontal body diameter, vertical body diameter, and head diameter against body volume and body length with P value < 0.05. Faint radiopaque images showed in the 2 fetus sample and strong radiopaque images showed in the 3 fetus sample. Radiopaque images were identified in the teeth, cranium, vertebrae, and extremities bones. In this study we concluded that there was a specific hair pattern in different fetal stage. All body measurements have positive correlation with body volume and body weight and x-ray analysis shown that the ossification of the bone was started to happen while the smooth hair was growth.
PubMed: 34691263
DOI: 10.1590/1984-3143-AR2021-0005 -
Anais Da Academia Brasileira de Ciencias 2023The present study aims to structurally and histochemically characterize the Gymnotus carapo tegument. 30 specimens were captured and slaughtered by spinal section with...
The present study aims to structurally and histochemically characterize the Gymnotus carapo tegument. 30 specimens were captured and slaughtered by spinal section with anesthesia. The observation was carried out with a stereoscopic microscope and the body surface was photographed. Fragments of the dorsal, ventral and lateral region were fixed in Bouin's solution for 12 hours and subsequently preserved in 70% alcohol. They were subsequently observed in the scanning electron microscope (SEM). The preparation for SEM was performed following the standardized protocol. Histological preparations were made, and the cuts were colored with H-E, PAS and Coomassie Blue. The images were obtained in an Olympus BX41-ENUTV-4 microscope. From the observations in SEM a plain tegument with pores of different sizes could be evidenced. The scales of the different regions of the body have different ornaments. Microscopically it was composed of a stratified non-keratinized epithelium consisting of two types of morphologically distinct cells: epidermal cells and mucous cells (PAS-Commassie Blue positive). Under the epithelium there is a layer of dense irregular connective tissue with associated chromatophores and more deeply scales. These analyzes are the basis for future studies that will focus on elucidating the events related to integumentary healing in this species.
Topics: Animals; Gymnotiformes; Fishes; Skin; Anesthesia; Ethanol
PubMed: 37729295
DOI: 10.1590/0001-3765202320191259 -
PloS One 2021This study aimed to investigate the effectiveness of leaf ethanolic extract of Etlingera hemisphaerica (LE3H) in reducing defects in fetal anatomy and endochondral...
Leaf ethanolic extract of Etlingera hemesphaerica Blume mitigates defects in fetal anatomy and endochondral ossification due to mercuric chloride during the post-implantation period in Mus musculus.
This study aimed to investigate the effectiveness of leaf ethanolic extract of Etlingera hemisphaerica (LE3H) in reducing defects in fetal anatomy and endochondral ossification in mice induced by HgCl2 during the post-implantation period. Pregnant mice were divided into four groups, each consisting of 10 dams, and received drink and food ad libitum. The first group was administered LE3H (E1), the second one HgCl2 (E2), the third one HgCl2+LE3H (E3), and the fourth was control (E0), administered double-distilled water only. HgCl2 (5 mg/kg bw) was administrated by injection intraperitoneally on gestation day (GD)9 and LE3H (0.39 mg/g bw) was administered by gavage on GD10. The treated and control animals were killed by cervical dislocation on GD18, dissected, and the morphologically normal living fetuses (MNLF) were collected. The MNLF of E0, E1, E2, and E3 from 5 dams were fixed with Bouin solution, and observed using the free hand razor blade technique for soft tissue examination. The remaining MNLF were fixed with 96% ethanol, and then stained with Alizarin Red S and Alcian Blue for ossification examination. Index of length of ossified part (ILOP) of humerus, index of width of ossified part (IWOP) of humerus, ILOP of femur, and IWOP of femur were calculated. E2 had higher cases of anatomical defects (74,6%) than E3 (48.9%), E1 (15.0%), and E0 (0%). E2 had humerus IWOP of 0.82±0.03, which was significantly lower than that of E0 (0.89±0.04) and E1 (0.89±0.03), while that of E1 and E0 was not significantly different from each other. Meanwhile, IWOP in E3 (0.88±0.03) was significantly higher than that in E2, but not different from that in E1 and E0. Thus, LE3H mitigated defects in fetal anatomy and endochondral ossification induced by HgCl2 in mice.
Topics: Animals; Embryo Implantation; Ethanol; Female; Fetal Diseases; Fetus; Injections, Intraperitoneal; Mercuric Chloride; Mice; Osteogenesis; Plant Extracts; Plant Leaves; Pregnancy; Zingiberaceae
PubMed: 33690696
DOI: 10.1371/journal.pone.0247467 -
Iranian Red Crescent Medical Journal Feb 2014Advancement in the treatment of various types of cancer has led to greater patient survival. These treatments essentially have toxic effects on different kinds of cells,...
BACKGROUND
Advancement in the treatment of various types of cancer has led to greater patient survival. These treatments essentially have toxic effects on different kinds of cells, such as germ cells. Infertility as one of the side effects of cancer treatment has changed the quality of life of young cancer survivors dramatically. Melatonin is an antioxidant with receptors in the reproductive systems.
OBJECTIVES
We supposed that melatonin, as an antioxidant, may protect testis against the toxic effects of the drugs.
MATERIALS AND METHODS
In this experimental study, three groups with seven mice each, were allocated. The control group received normal saline for two months, and the busulfan group received a single dose of 40 mg/kg busulfan intra-peritoneally, and the melatonin group received 20 mg/kg melatonin daily for two months, 45 days after a single dose of busulfan. Next, after decapitation and removal of the testis, tissues were fixed in Bouin's solution and stained by H&E and TUNEL. The sections were evaluated, assessing morphology and spermatogenesis.
RESULTS
In this research, a significant reduction in Johnson's criteria in the busulfan group (Mean rank = 15.50) was found versus the control group (Mean rank = 45.50), P < 0.001 and in the melatonin group (Mean rank = 45.50) compared to the busulfan group (Mean rank = 15.50), P < 0.001. There was a significant difference between the melatonin and control groups, P < 0.05. In addition, a significant decrease in seminiferous tubule diameter was observed in the busulfan group (763.2 ± 104.41) versus the control group (855.4 ± 52.35), P < 0.01 and melatonin group (834.2 ± 87.26), P < 0.05. Testicular epithelium height was significantly decreased in the busulfan group (Mean rank = 14.60) compared to the control group (Mean rank = 26.40), P < 0.01 and in the busulfan group (Mean rank = 14.95) in comparison with the melatonin group (Mean rank = 26.05), P < 0.01. Also melatonin group (Mean rank = 25.42) showed a significant reduction in epithelium height compared to the control group (Mean rank = 35.58), P < 0.05. Spermatogenesis was impaired in the busulfan group. Although melatonin reduced the rate of apoptosis in the busulfan group, yet it could not remove all apoptotic cells.
CONCLUSIONS
This study indicated that melatonin ameliorates the cytotoxic effects of busulfan on germ cells.
PubMed: 24719743
DOI: 10.5812/ircmj.14463 -
Heliyon Mar 2023The present study aimed to investigate the effect of co-exposure to heat and psychological stressors on testicular tissue as one of the major male reproductive organs in...
OBJECTIVES
The present study aimed to investigate the effect of co-exposure to heat and psychological stressors on testicular tissue as one of the major male reproductive organs in rats.
METHODS
Forty adult male Wistar rats were divided into four groups of 10 animals each. The first group was exposed to heat stress (Temperature Humidity Index: 57.75 ± 3), the second group was exposed to three psychological stressors including strobe light ultrasonic sound, and tilting cage, and the third group was co-exposed to both heat and psychological stress daily. The order of exposure to various psychological stressors was randomized. Following the last day of the 40 -day exposure, the rats were euthanized and their testicular tissues were fixed in Bouin's solution. Moreover, a tissue processor, a microtome as well as hematoxylin, and eosin staining were used for tissue preparation.
RESULTS
The co-exposure to psychological and heat stress can cause negative testis histopathological changes including spermatid retention, degeneration of round spermatids and spermatocytes, degeneration and depletion of elongated Spermatid, Sertoli cell status, tubular degeneration/atrophy, Leydig cell atrophy, tubular dilatation, re-tubular dilation, and sclerosis status in a number of rats. Moreover, the histopathological changes were higher in the group exposed to heat stress compared with those exposed to psychological stress.
CONCLUSIONS
Although exposure to either stressor alone can affect testicular tissue as part of the reproductive system, co-exposure to both stressors may result in an increased risk of adverse effects on testicular tissue.
PubMed: 36915529
DOI: 10.1016/j.heliyon.2023.e14146 -
International Journal of Fertility &... Oct 2011di (2-ethylhexyl) phthalate (DEHP) is widely used in the plastic industry and can induce reproductive toxicity. On the other hand, L-carnitine (LC) plays a crucial role...
BACKGROUND
di (2-ethylhexyl) phthalate (DEHP) is widely used in the plastic industry and can induce reproductive toxicity. On the other hand, L-carnitine (LC) plays a crucial role in sperm metabolism and maturation. This study evaluates the effect of LC on body and testis weight, testis tissue, count, motility, viability, morphology, and chromatin quality of epididymal sperm, testicular spermatid number (TSN) per gram testis and daily sperm production (DSP) in LC-treated mice.
MATERIALS AND METHODS
IN THIS EXPERIMENTAL STUDY, ADULT MALE NMRI MICE (MEAN AGE: 4 weeks) were given doses of DEHP and LC by gavaging for 2 weeks. All samples were assessed according to World Health Organization (WHO) criteria. Sperm morphology was assessed using Papanicolaou staining and sperm chromatin quality by aniline-blue staining. The left testes were fixed in Bouinś solution for histological examination and the end slices were stained with hematoxylin and eosin (H&E). The right testes were homogenized, and then TSN and DSP were calculated with an improved Neubauer haemocytometer and respective frames. Paired t-test, ANOVA, and Kruskal-Wallis tests were utilized for data analysis.
RESULTS
Co-administration of DEHP and LC not only prevented significant gains in testicular weight, but also maintained the sperm's normal morphology and chromatin quality (p<0.05). In addition, LC recovered histological changes, TSN, DSP, and sperm count.
CONCLUSION
These results demonstrated that oral administration of LC partially or generally protects spermatogenesis from DEHP-toxicity in mice.
PubMed: 25101163
DOI: No ID Found -
The Journal of Histochemistry and... Jan 2020Hyaluronan (HA) is a ubiquitous component of the extracellular matrix. The spatial-temporal localization of HA can be visualized in situ using biotinylated HA binding...
Hyaluronan (HA) is a ubiquitous component of the extracellular matrix. The spatial-temporal localization of HA can be visualized in situ using biotinylated HA binding proteins (HABPs). This assay is sensitive to fixation conditions, and there are currently no best practices for HA detection. Thus, the goal of this study was to optimize fixation conditions for visualizing HA in the ovary, kidney, and liver through analysis of six commonly used fixatives for HA detection: Bouin's Solution, Carnoy's Solution, Ethanol-Formalin-Glacial Acetic Acid (EFG), Histochoice, Modified Davidson's Solution, and 10% Neutral Buffered Formalin. Organs were harvested from CB6F1 mice and fixed with one of the identified fixatives. Fixed organs were sectioned, and the HABP assay was performed on sections in parallel. Hematoxylin and eosin staining was also performed to visualize tissue architecture. HABP signal localization and intensity varied between fixatives. EFG and Carnoy's Solution best preserved the HA signal intensity in the ovary and liver, showing HA localization in various sub-organ structures. In the kidney, only Modified Davidson's Solution was less than optimal. Our findings demonstrate that fixation can alter the ability to detect HA in tissue macro- and microstructures, as well as localization in a tissue-specific manner, in situ.
Topics: Animals; Female; Humans; Hyaluronic Acid; Kidney; Liver; Mice; Organ Specificity; Ovary; Rats; Tissue Embedding; Tissue Fixation
PubMed: 31714169
DOI: 10.1369/0022155419884879 -
Andrology Jan 2018Histone H3.5 (H3.5) is a newly identified histone variant highly expressed in the human testis. We have reported the crystal structure, instability of the H3.5...
Histone H3.5 (H3.5) is a newly identified histone variant highly expressed in the human testis. We have reported the crystal structure, instability of the H3.5 nucleosome and accumulation around transcription start sites, mainly in primary spermatocytes, but its role in human spermatogenesis remains poorly understood. Testicular biopsy specimens from 30 men (mean age: 35 years) with non-obstructive azoospermia (NOA) who underwent microdissection testicular sperm extraction and 23 men with obstructive azoospermia (OA) were included. An H3.5-specific mouse monoclonal antibody recognizing an H3.5-specific synthetic peptide was generated, and immunohistological staining for H3.5 and proliferating cell nuclear antigen (PCNA) was performed on Bouin's solution-fixed sections. Expression and localization of H3.5 were compared with patient background, germinal stage, and PCNA expression. In testes of patients with normal spermatogenesis, differentially expressed H3.5 was specifically localized in either spermatogonia or preleptotene/leptotene-stage primary spermatocytes, especially during germinal stages VI-X. In NOA testes, mRNA expression of H3.5 (H3F3C) was significantly reduced compared with other H3 histone family members, and expression of H3.5 was significantly lower than that in OA. Additionally, the number of H3.5-positive germ cells was higher in hypospermatogenesis or late maturation arrest than in early maturation arrest in NOA testes (p < 0.01). A significant positive correlation was observed between H3.5 and PCNA expression (p < 0.05) but not TUNEL-positive cells, and expression of H3.5 was enhanced after hCG-based salvage hormonal therapy. Different from other testis-specific histones, which are often expressed during the histone-to-protamine transition during meiosis, H3.5 was expressed mainly in immature germ cells. H3.5 may play roles in DNA synthesis, but not apoptosis, and its expression is regulated by gonadotropins, indicating that such epigenetic regulations are important in normal spermatogenesis and spermatogenic disorders.
Topics: Adult; Azoospermia; Histones; Humans; Male; Middle Aged; Retrospective Studies; Spermatogenesis; Spermatozoa
PubMed: 29179259
DOI: 10.1111/andr.12438 -
Journal of Clinical and Diagnostic... Aug 2016Preparation of good tissue specimens for microscopy requires complete fixation. No ideal fixative has been found till date, with every fixative showing advantages and...
INTRODUCTION
Preparation of good tissue specimens for microscopy requires complete fixation. No ideal fixative has been found till date, with every fixative showing advantages and disadvantages. Appropriate fixation is required to maintain clear and consistent morphologic features for histologic examination. Pathologists mostly examine formalin fixed tissue sections and are less used to the morphologic changes induced by other fixatives. Underfixed and overfixed tissue in various fixatives can lead to tissue architectural changes which can affect its diagnostic value.
AIM
To assess sectioning ability, staining intensity and microscopic details of tissues kept in different fixatives at different time intervals.
MATERIALS AND METHODS
Fresh tissue specimen i.e., goat tongue was collected and its middle-third portion was used for the study purpose. The tissue was grossed into 10 equal pieces and kept in various fixatives (10% Buffered formalin, Carnoy's solution, Absolute ethyl alcohol, Bouin's fluid) for five different time intervals (6, 12, 18, 24 and 30 hours) and normal tissue processing steps were carried out followed by sectioning and staining. During sectioning, sectioning parameter was assessed. Following sectioning, sections were observed under light microscope and were histologically evaluated for staining and microscopic details. To calculate the sectioning parameter Fisher's exact test was used and to assess parameters for staining and microscopic details Mann-Whitney U test was used.
RESULTS
According to the study, 10% buffered formaldehyde is considered as a superior fixative under all parameters followed by Bouin's fluid, Carnoy's solution and Absolute alcohol.
CONCLUSION
In our study, it was concluded that 10% buffered formaldehyde should be continued as a routine fixative however, other fixatives can be used depending upon the non-availability of required fixative or in case of emergencies. Pathologist should be accustomed to histologic and morphologic changes of underfixed and overfixed tissue which can affect its diagnostic value.
PubMed: 27656558
DOI: 10.7860/JCDR/2016/19007.8236 -
Fish Physiology and Biochemistry Jun 2021Sharks are not only threatened, but also have a low fecundity as they are being overfished. The shark family, Sphyrnidae, consists of nine species of which three are...
Sharks are not only threatened, but also have a low fecundity as they are being overfished. The shark family, Sphyrnidae, consists of nine species of which three are found in South African oceans. One of the three Sphyrnidae species, the scalloped hammerhead (Sphyrna lewini) are the most common, but their biology and mode of reproduction are not extensively studied in terms of their reproductive biology. The aim of this study was to describe the germ cell development in the testes of sexually mature male scalloped hammerheads. Three individual male S. lewini were caught at Zinkwazi, KwaZulu-Natal, South Africa. The sharks and their reproductive organs were weighed and measured to collect the biometric data for the condition factor and the gonado-somatic index. Following standard necropsy, the testes were fixed in Bouin's solution and processed for histological assessment. The histological assessment revealed that the testes of S. lewini consist of seminiferous tubules which form part of a larger lobular structure with germ cells in different stages of development, from spermatogonia to mature spermatozoa. Seven stages of development were identified during the process of spermatogenesis, similar to what has been described for elasmobranchs. In conclusion, this study provides evidence that the testes of S. lewini are diametrical and polyspermatocystic and conforms to the testes structure of elasmobranch males.
Topics: Animals; Male; Sharks; South Africa; Spermatogenesis; Testis
PubMed: 32915423
DOI: 10.1007/s10695-020-00871-z