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Theriogenology Jul 2020The objective of this review is to provide new insights into the possible use of a proteomic method known as Intact Cell Matrix-Assisted Laser Desorption-ionization... (Review)
Review
The objective of this review is to provide new insights into the possible use of a proteomic method known as Intact Cell Matrix-Assisted Laser Desorption-ionization Time-Of-Flight Mass Spectrometry (ICM-MS) in animal clinical research. Here, we give an overview of the basics of this technique, its advantages and disadvantages compared with other proteomic approaches, past applications and future perspectives. A special emphasis on its implementation in animal reproduction science is given, including examples of the reliable use of ICM-MS on fertility screening. In mammals, the ICM-MS profiles from pig epididymal spermatozoa reflect the proteome changes that they undergo during epididymal maturation and could be associated with the acquisition of fertilizing ability. In chicken, using adequate pre-processing and bioinformatics analysis tools, sperm ICM-MS profiles showed characteristic spectral features that allowed their classification according to their actual fertilizing ability. The association of ICM-MS and Top-down proteomic strategies allowed the identification of chicken fertility biomarkers candidates such as protein vitelline membrane outer layer protein 1 (VMO-1) and avian beta-defensin 10 (AvBD10). In female reproduction, a similar approach on ovarian follicular cells allowed the identification of specific markers of oocyte maturation in the oocyte and surrounding cumulus cells. Altogether, these results indicate that ICM-MS profiling could be a suitable approach for molecular phenotyping of male and female gametes.
Topics: Animals; Gene Expression Regulation; Livestock; Proteomics; Reproduction; Single-Cell Analysis; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
PubMed: 32284210
DOI: 10.1016/j.theriogenology.2020.02.037 -
Animals : An Open Access Journal From... Feb 2023This study evaluates the effect of housing environment on the egg quality characteristics of brown egg layers as many different environments are currently used in the...
This study evaluates the effect of housing environment on the egg quality characteristics of brown egg layers as many different environments are currently used in the industry. Battery cages, barren colony cages, enriched colony cages, cage-free, and free-range environments were evaluated. Overall, all egg quality measurements were affected by housing environment ( < 0.01) except for vitelline membrane strength, elasticity, and egg solids. Eggshells and yolks were lightest in barren colony cages and darkest from free-range hens ( < 0.0001). Free-range eggs were heavier than eggs from all other environments ( < 0.0001). Cage-free eggs had lower albumen height and Haugh units than other environments ( < 0.0001). Lastly, cage-free and free-range eggs had stronger eggshells than the other environments ( < 0.0001), and free-range eggs had more elastic eggshells than eggs from conventional battery cages and barren colony cages ( < 0.01). Access to the range seemed to give free-range hens different nutritional advantages, which allowed for the darker yolks and shells. Furthermore, eggs from barren colony cages seemed to exhibit more negative characteristics. Simply adding enrichments to colony cages did not improve or detract from egg quality. From this research, it appears that, as the industry moves toward extensive environments, the egg quality of brown egg layers will improve.
PubMed: 36830504
DOI: 10.3390/ani13040716 -
Scientific Reports Jan 2019In the ascidian Ciona robusta (formerly C. intestinalis type A), the mechanism underlying sperm penetration through the egg investment remains unknown. We previously...
In the ascidian Ciona robusta (formerly C. intestinalis type A), the mechanism underlying sperm penetration through the egg investment remains unknown. We previously reported that proteins containing both an astacin metalloprotease domain and thrombospondin type 1 repeats are abundant in the sperm surface protein-enriched fraction of C. robusta. Here we investigated the involvement of those proteins in fertilisation. We refined the sequences of astacin metalloproteases, confirmed that five of them are present in the sperm, and labelled them as tunicate astacin and thrombospondin type 1 repeat-containing (Tast) proteins. Fertilisation of C. robusta eggs was potently inhibited by a metalloprotease inhibitor GM6001. The eggs cleaved normally when they were vitelline coat-free or the inhibitor was added after insemination. Furthermore, vitelline coat proteins were degraded after incubation with intact sperm. These results suggest that sperm metalloproteases are indispensable for fertilisation, probably owing to direct or indirect mediation of vitelline-coat digestion during sperm penetration. TALEN-mediated knockout of Tast genes and the presence of GM6001 impaired larval development at the metamorphic stage, suggesting that Tast gene products play a key role in late development.
Topics: Animals; Ciona intestinalis; Egg Proteins; Female; Male; Metalloproteases; Sperm-Ovum Interactions; Spermatozoa; Vitelline Membrane
PubMed: 30700775
DOI: 10.1038/s41598-018-37721-1 -
Open Biology Sep 2022During early avian development, only a narrow band of cells (the edge cells, also called 'margin of overgrowth') at the rim of the embryo is responsible for blastoderm...
During early avian development, only a narrow band of cells (the edge cells, also called 'margin of overgrowth') at the rim of the embryo is responsible for blastoderm expansion by crawling over the vitelline membrane (VM) to cover the whole egg yolk in just 4 days (a process called epiboly). Surprisingly, this has not yet been studied in detail. Here we explore the edge cells of the chick embryo using hybridization, immunohistochemistry and live imaging. Morphological and molecular properties reveal that the edge has a distinctive structure, being subdivided into sub-regions, including at least four distinct zones (which we name as leading, trailing, deep and stalk zones). This allows us to study reorganization of the edge region that accompanies reattachment of an explanted blastoderm to the VM. Immunohistochemistry uncovers distinct polarized cellular features resembling the process of collective cell migration described in other systems. Live imaging reveals dynamic lamellipodial and filopodial activity at the leading edge of the outermost cells. Our data provide evidence that edge cells are a distinct tissue. We propose that edge cells may be a useful model system for the study of wound healing and other closure events in epithelial cell sheets.
Topics: Animals; Blastoderm; Cell Movement; Chick Embryo; Epithelial Cells; Vitelline Membrane; Wound Healing
PubMed: 36128719
DOI: 10.1098/rsob.220147 -
Applied Microscopy Jul 2022We examined the morphology of the fertilized egg and the fine structure of fertilized egg envelopes of Poropanchax normani belonging to the family Poeciliidae, also...
We examined the morphology of the fertilized egg and the fine structure of fertilized egg envelopes of Poropanchax normani belonging to the family Poeciliidae, also known as Norman's lampeye using light and electron microscopes. The fertilized eggs with narrow perivitelline space were found to be spherical and demersal, additionally containing small oil droplets in the vitelline membrane. Further, a bundle of adhesive filaments was observed to be present on one side of the fertilized egg. These filaments possessed remarkably high elasticity and were approximately 1-3 mm in length. The size of the fertilized egg was determined to be about 1.49 ± 0.07 mm (n = 30). The outer surface appeared smooth, and adhesive filaments originating at different location of the surface of the envelope were found to be distributed around the egg envelope and were joined together to form a single long bundle in scanning electron microscopic observation. A peak-like structure formed of several straight wrinkles was observed around the micropyle. However, the complete structure of the micropyle could not be studied due to the depth at which it was located. Additionally, the total thickness of the egg envelope was ascertained to be approximately12.5-14.5 μm. The egg envelope consisted of two distinct layers, an outer electron dense layer and an inner lamellar layer, further consisting of 10 sublayers of varying thicknesses. Collectively, it was observed that the morphological characteristics of the fertilized egg, fine structures surrounding the micropyle, outer surface, adhesive structure consisting adhesive filaments, and sections of fertilized egg envelope displayed species specificity.
PubMed: 35831688
DOI: 10.1186/s42649-022-00075-0 -
Journal of Food Science May 2022Herein, the water and lipid migration of salted duck eggs during storage were systematically explored in three different packaging conditions of long-term salting, no...
Herein, the water and lipid migration of salted duck eggs during storage were systematically explored in three different packaging conditions of long-term salting, no packaging, and vacuum packaging. Bound water, multilayer bound water, lipid, and bulk water were observed in the whole duck egg by low-field nuclear magnetic resonance (LF-NMR) relaxation. Five weeks of salting process led to the redistribution of water and lipid due to the watery state of egg white and the gelation of egg yolk due to the permeation of salt, and boiling mainly caused an obvious decrease in the mobility of bulk water due to the gelation of egg white. Among these three conditions, long-term salting with 6 months storage caused the most serious redistribution of water and lipid as well as the rupture of the vitelline membrane, but could prevent the oxidation of egg yolk. Vacuum packaging had the least influence on the water and lipid distribution, mass change, and water content but led to lipid oxidation with high degree in egg yolk. However, the most obvious mass loss was observed in the salted duck eggs during the storage without packaging. In addition, principal component analysis of Carr-Purcell-Meiboom-Gill data suggested that LF-NMR could distinguish the salted duck eggs with different storage times during the early stage of the storage. Practical Application Water and lipid migration of salted duck eggs during storage with three packaging conditions were explored by using low-field nuclear magnetic resonance and magnetic resonance imaging. Understanding the impacts of packaging conditions on water and lipid migration of salted duck eggs during storage could provide a new method for the quality identification.
Topics: Animals; Ducks; Egg Yolk; Eggs; Lipids; Magnetic Resonance Imaging; Magnetic Resonance Spectroscopy; Sodium Chloride; Water
PubMed: 35411557
DOI: 10.1111/1750-3841.16139 -
Recent Patents on Anti-infective Drug... 2017Triamcinolone is a long acting corticosteroid used in the treatment of arthritis, eczema, psoriasis and similar conditions which cause inflammation. Triamcinolone has...
BACKGROUND
Triamcinolone is a long acting corticosteroid used in the treatment of arthritis, eczema, psoriasis and similar conditions which cause inflammation. Triamcinolone has half-life of 88min. Prolonged oral use is associated with gastrointestinal adverse effects as peptic ulcer, abdominal distention and ulcerative esophagitis as described in various patents. Microemulgel offers advantage of better stability, better loading capacity and controlled release especially for drug with short half life.
OBJECTIVE
Objective of the present study was to optimize microemulgel based transdermal delivery of triamcinolone.
METHOD
Saturated solubility of triamcinolone in various oils, surfactants and co-surfactants is estimated. Pseudo-ternary phase diagrams were constructed to determine the region of transparent microemulsion. Microemulsion was evaluated for globule size (FE-SEM, zetasizer), % transmittance, pH, viscosity, conductivity etc. Design of experiment was used to optimize microemulsion based gel. Carbopol 971P and HPMC K100M were used as independent variables. Microemulsion based gel was evaluated for in-vitro as well as ex-vivo parameters.
RESULTS
Microemulsion was formulated with oleic acid, lauroglycol FCC and propylene glycol. PDI 0.197 indicated microemulsion is mono-disperse. 32 factorial design gave batch F8 as optimized. Design expert suggested drug release; gel viscosity and bio-adhesive strength were three significant dependant factors affecting the transdermal delivery. F8 showed drug release 92.62.16±1.22% through egg membrane, 95.23±1.44% through goat skin after 8hr and Korsmeyer-Peppas release model was followed.
CONCLUSION
It can be concluded that a stable, effective controlled release transdermal microemulgel was optimised for triamcinolone. This would be a promising tool to deliver triamcinolone with enhanced bioavailability and reduced dosing frequency.
Topics: Administration, Cutaneous; Animals; Arthritis; Delayed-Action Preparations; Drug Liberation; Eczema; Emulsions; Gels; Glucocorticoids; Goats; Half-Life; Models, Chemical; Oils; Particle Size; Patents as Topic; Permeability; Psoriasis; Skin; Solubility; Surface-Active Agents; Triamcinolone; Vitelline Membrane
PubMed: 28506203
DOI: 10.2174/1574891X12666170426092911 -
Veterinary Microbiology May 2020In 2019, a novel goose astrovirus (GoAstV) epidemiological investigation on geese was conducted in Shandong province, China. During the investigation, a high prevalence...
In 2019, a novel goose astrovirus (GoAstV) epidemiological investigation on geese was conducted in Shandong province, China. During the investigation, a high prevalence of novel GoAstV was observed in symptom-free breeding geese flocks. Moreover, the novel GoAstV-specific RNA was detected in either breeder birds or their progenies. To verify the hypothesis that the novel GoAstV could be transmitted vertically, a total of 42 WuLong breeder geese, aged 335 days, were equally divided into three groups for experimental infection. The SDPY isolate of novel GoAstV (A/goose/Shandong/SDPY/2018, SDPY), preserved in our laboratory, was injected intramuscularly to subjects of group A while orally inoculated to those of group B. After the inoculation, novel GoAstV RNA was detected in vitelline membrane, embryos, and allantoic fluid of goose embryos in novel GoAstV infected groups. Moreover, the ORF2 gene of novel GoAstV from vitelline membrane, embryo, allantoic fluid as well as gosling shared almost 100 % nucleotide homology to a novel GoAstV virus isolated from the goose ovary which produced the egg, suggesting that the novel GoAstV can be vertically transmitted in the goose. Taken together, the findings provide evidence of possible vertical transmission of novel GoAstV from breeding goose to goslings.
Topics: Animals; Astroviridae; Astroviridae Infections; China; Female; Geese; Infectious Disease Transmission, Vertical; Male; Phylogeny; Poultry Diseases; RNA, Viral
PubMed: 32402337
DOI: 10.1016/j.vetmic.2020.108657 -
Biology Open Sep 2019Axis specification is a fundamental developmental process. Despite this, the mechanisms by which it is controlled across insect taxa are strikingly different. An...
Axis specification is a fundamental developmental process. Despite this, the mechanisms by which it is controlled across insect taxa are strikingly different. An excellent example of this is terminal patterning, which in Diptera such as occurs via the localized activation of the receptor tyrosine kinase Torso. In Hymenoptera, however, the same process appears to be achieved via localized mRNA How these mechanisms evolved and what they evolved from remains largely unexplored. Here, we show that , known for its role in terminal patterning, is instead required for the integrity of the vitelline membrane in the hymenopteran wasp We find that other genes known to be involved in terminal patterning, such as and , also do not function in embryonic development. These findings extended to orthologues of vitelline membrane proteins known to play a role in localizing Torso-like in ; in these are instead required for dorso-ventral patterning, gastrulation and potentially terminal patterning. Our data underscore the importance of the vitelline membrane in insect development, and implies phenotypes caused by knockdown of must be interpreted in light of its function in the vitelline membrane. In addition, our data imply that the signalling components of the terminal patterning systems were co-opted from roles in regulating moulting, and co-option into terminal patterning involved the evolution of a novel interaction with the vitelline membrane protein Torso-like.This article has an associated First Person interview with the first author of the paper.
PubMed: 31488408
DOI: 10.1242/bio.046284 -
Biomedical Reports Mar 2019Several potential urinary biomarkers exhibiting an association with upper gastrointestinal tumour growth have been previously identified, of which S100A6, S100A9,...
Several potential urinary biomarkers exhibiting an association with upper gastrointestinal tumour growth have been previously identified, of which S100A6, S100A9, rabenosyn-5 and programmed cell death 6-interacting protein (PDCD6IP) were further validated and found to be upregulated in malignant tumours. The cancer cohort from our previous study was subclassified to assess whether distinct molecular markers can be identified for each individual cancer type using a similar approach. Urine samples from patients with cancers of the stomach, oesophagus, oesophagogastric junction or pancreas were analysed by surface-enhanced laser desorption/ionization-time-of-flight mass spectrometry using both CM10 and IMAC30 (Cu-complexed) chip types and LC-MS/MS-based mass spectrometry after chromatographic enrichment. This was followed by protein identification, pattern matching and validation by western blotting. We found 8 m/z peaks with statistical significance for the four cancer types investigated, of which m/z 2447 and 2577 were identified by pattern matching as fragments of cathepsin-B (CTSB) and cystatin-B (CSTB); both molecules are indicative of pancreatic cancer. Additionally, we observed a potential association of upregulated α-1-antichymotrypsin with pancreatic and gastric cancers, of PDCD6IP, vitelline membrane outer layer protein 1 homolog (VMO1) and triosephosphate isomerase (TPI1) with oesophagogastric junctional cancers, and of complement C4-A, prostatic acid phosphatase, azurocidin and histone-H1 with oesophageal cancer. Furthermore, the potential pancreatic cancer biomarkers CSTB and CTSB were validated independently by western blotting. Therefore, the present study identified two new potential urinary biomarkers that appear to be associated with pancreatic cancer. This may provide a simple, non-invasive screening test for use in the clinical setting.
PubMed: 30906545
DOI: 10.3892/br.2019.1190