Layer Of Cytoplasm Research Articles

7097 The role of mitochondrial fraction locating cytoplasmic and peridroplet area in white adipocyte

Abstract Disclosure: I. Mori: None. T. Ishizuka: None. H. Morita: None. K. Kajita: None. White adipocytes (WAs) are involved in the formation of obesity and insulin resistance, but little is known about the role of WAs mitochondria. A recent study revealed that the mitochondria of brown adipocytes (BAs) consist of cytoplasmic mitochondria CM), which is responsible for β-oxidation, and peridroplet mitochondria (PDM), which supplies triglyceride to lipid droplets. However, it is unclear whether such a functional division exist in WA mitochondria. In this study, we examined whether the pellets obtained from cytoplasmic and oil layers of WAs of C57/BL/J mice by centrifugation were mitochondria. Since mitochondrial DNA, mitochondrial protein, Mitotracker Red staining, and mitochondrial images in electron microscope were detected in both fractions, they were considered to be cytoplasmic mitochondria (CMw) and peridroplet mitochondria (PDMw) in WAs, respectively. The amount of CMw and PDMw were greater in epididymal fat than in inguinal fat. CMw had higher β-oxidation activity than PDMw. PDMw was associated with perilipin 1/3 and diacylglycerol acyltransferase 2. The amount of PDMw was elevated in epididymal fat of mice fed a high-fat diet (HFD) for 2 and 4 weeks compared to normal chew-fed mice. These results suggested that CMw was involved in β-oxidation, and PDMw in droplet expansion. Although numerous researches have been shown that amount of mitochondria is suppressed in obese WAs, our results revealed that PDMw might be involved in the early stage of obesity formation. Presentation: 6/1/2024

Pleomorphic Adenoma of the Upper Lip: A Case Report

Pleomorphic adenoma (PA) is a commonly occurring benign tumor originating in the salivary glands. The incidence is mostly in the parotid glands (85%) followed by minor salivary glands (10%). Pleomorphic adenoma from minor salivary glands of the lips is a rare neoplasm. Here, we present a case of a patient with PA in the upper lip, reporting a common neoplasm in a rare site. A 82-year-old male with a slowly progressing swelling that occurred 20years ago on the upper white lip. This tumor is not associated with pain and tenderness. The mass was about 1.5x1.5cm sized, circumscribed, sessile and firm on the external upper lip without oral side bulging (Fig. 1). The tumor was removed completely with a elliptical incision under local anesthesia. The encapsulated mass was measured 0.9x0.9x0.6cm. On histology, a well-circumscribed growth was seen comprising epithelial and stromal components. The epithelial component formed glandular structures lined by round-to oval cells having small nuclei, pink cytoplasm and myoepithelial basal cell layer while the stroma had a fibromyxoid appearance(Fig. 2). Postoperatively wound was well healed without recurrence after 6months. Pleomorphic adenoma of the upper lip is a rare neoplasm, especially not intraoral growth. For aesthetic good result, the mass was removed without hesitation. It is required a high index of suspicion and a long-term follow-up because it could recur and rarely be transformed into malignancy.

Comparative analysis of the cytoarchitecture of the excretory bladder of adult Digenea (Platyhelminthes) with consideration of the presence of mineralized excretory corpuscles in marine and freshwater adult worms

The ultrastructural differences are shown between the cytoarchitecture of the excretory bladder and excretory inclusions in four digenean species, two azygiids, the marine Otodistomum cestoides and the freshwater Azygia lucii, the marine derogenid Derogenes varicus and the freshwater allocreadiid Acrolichanus auriculatus. The unusual cytoarchitecture of the bladder epithelium of the azygiid digeneans, consisting along its entire length of two alternating, morphologically different zones, tegumental and cellular excretory epithelial zones, connected by septate junctions, has recorded for the first time for the Digenea and, in general, for the Neodermata. It, possible, suggests the participation of the tegumental distal cytoplasmic layer in the formation of their excretory bladder epithelium. Like most digeneans, the excretory bladder of A. auriculatus and D. varicus has a syncytial epithelial lining. Based on available literature and our own results, we can confirm the presence of the excretory corpuscles in adult marine digeneans and their absence from freshwater species, regardless of the digenean localization in their host. The present study shown that in marine digeneans, the excretory corpuscles are associated with specialized excretory cells or excretory syncytial epithelium. Ultrastructural data were obtained on the possible growth of the excretory bladder epithelium due to the migration of undifferentiated cells into the epithelial lining in studied marine species. We may assume that the bladder epithelium of marine adult digeneans specializes, in addition to the excretory function, in osmoregulatory function.

Milk fat globule membrane: formation and transformation.

The milk fat globule membrane (MFGM) is formed by complex cell biological processes in the lactating mammary epithelial cell which result in the release of the milk fat globule (MFG) into the secretory alveolus. The MFG is bounded by a continuous unit membrane (UM), separated from the MFG lipid by a thin layer of cytoplasm. This unique apocrine secretion process has been shown in all of the mammary species so far investigated. Once the MFG is released into the alveolus there is a considerable transformation of the UM with its attached cytoplasm. This is the MFGM. The transformation is stable and expressed milk shows the same transformed MFGM structure. Again, this transformation of structure is common to all mammalian species so far investigated. However, the explanation of the transformation very much depends on the method of investigation. Transmission electron microscope (TEM) studies suggest a literal breakdown to a discontinuous UM plus cytoplasm in patches and strands, whereas more recent confocal laser scanning light microscopy (CLSM) studies indicate a separation, in a continuous UM, of two phases, one liquid ordered and the other liquid disordered. This review is designed to show that the TEM and CLSM results show different views of the same structures once certain deficiencies in techniques are factored in.

Migration and ultrastructure of the acanthocephalan Echinorhynchus gadi Zoega in Müller, 1776 in intermediate host under experimental conditions.

Amphipods Eogammarus tiuschovi were experimentally infected by the acanthocephalan Echinorhynchus gadi (Acanthocephala: Echinorhynchidae). Within the first four days post-infection, acanthors of the acanthocephalan caused the cellular response of the host, which ended with their complete encapsulation on day 4 post-infection. The acanthors obtained through the experiment were examined ultrastructurally. Two syncytia (frontal and epidermal) and a central nuclear mass are found in the acanthor's body. The frontal syncytium has 3-4 nuclei and contains secretory granules with homogeneous, electron-dense contents. Since the secretory granules occupy only the anterior one-third of this syncytium, it is suggested that the contents of these granules are involved in the acanthor's migration through the gut wall of the amphipod. The central nuclear mass consists of an aggregation of fibrillar bodies and a few electron-light nuclei distributed on the periphery. Some of these nuclei, located near the central nuclear mass, are assumed to be a source of the acanthocephalan's internal organs. The epidermal syncytium surrounds the frontal syncytium and the central nuclear mass. It is represented by a superficial cytoplasmic layer, but the bulk of the cytoplasm is concentrated in the posterior one-third of the acanthor's body. Syncytial nuclei are evenly distributed throughout the cytoplasm. The muscular system of the acanthors consists of 10 longitudinal muscle fibers located below the superficial cytoplasmic layer and two muscle retractors crossing the frontal syncytium.

Mammary lipid secretion: a reassessment.

Mammary lipid secretion is generally held to be unique and remarkably uniform between the many different orders of mammals. It produces a unit membrane-bounded milk fat globule (MFG). The unit membrane is separated from the lipoprotein boundary of what was the cytoplasmic lipid droplet (CLD) boundary by a uniform layer of cytoplasmic proteins. In 3-8% of the MFG in all species examined this cytoplasmic layer widens to include cytoplasmic organelles which are referred to as 'crescents'. This defines the MFG secretion as apocrine indicating a closely regulated process which minimises the loss of mammary epithelial cell (MEC) cytoplasm. The apocrine nature of the secretion might be expected since the evolution of the mammary gland is considered to be from an apocrine secreting skin gland. This short Research Reflection review is designed to investigate the exact cytoplasmic interactions which allow such efficient lipid secretion. There are two main scenarios: one which assumes that the observed close association between CLD and GV results in the CLD being released as a consequence of sequential exocytosis of the content of the associated GV. The second assumes that the CLD and the MEC apical plasmalemma interact in some way which causes the CLD to rise out of the cytoplasm enveloped in the plasmalemma. Here I present the evidence for the two possibilities. The first scenario is favoured, but the second cannot be ruled out.

Impact of Water Deficit Condition on Osmoregulation of the Brassica Species

Oilseed rape and mustard are mostly grown on conserved soil water in the Indian sub-continent. These crops generally suffer from water stress at reproductive period of development. Thus, to obtain higher stable yields of Brassica species under routine stress conditions, it is essential to identify and understand the interactions of different morpho-physiological traits responsible for drought resistance. To explicate interaction of traits related to biochemical, physio-morphological factors for sustaining drought resistance in Brassica species. Dry mass production and partition in a plant is important when increased yield are sought. A osmometer like plant cell which allows only selective solutes to pass through elastic membrane, cell wall and the thin layer of cytoplasm and a vacuole containing an aqueous solution. To express drought tolerance turgidity of cells maintained by osmotic adjustments.

Vampyrophrya pelagica (Chatton and Lwoff 1930), an apostome ciliate infectious on calanoid copepod from the Northeastern Arabian Sea

BackgroundZooplankton provides a dynamic habitat to various kinds of epibionts in the marine ecosystem and also forms a source of food and nutrients to various parasites and parasitoids. The study was conducted to identify the association of ciliates with calanoid copepod of Northeastern Arabian Sea. Zooplankton samples collected were analysed under stereomicroscope for any kind of ciliate attachments. The infective stage and major characteristics of the ciliates were discussed. The present observation is the first report of apostome ciliate Vampyrophrya pelagica on calanoid copepod Bestiolina similis from the open ocean waters off Tarapur, Northeastern Arabian Sea, during the early spring intermonsoon season and also a new host record for the ciliate.ResultsAnalysis of ~ 2530 copepod revealed the presence of 12 copepod specimens infested with apostome ciliate Vampyrophrya pelagica. All the host copepod, identified as Bestiolina similis, were females. Approximately 50–60 ciliates occupied the host endocuticular region. V. pelagica was represented by fully grown trophont stage characterized by the presence of a large food vacuole surrounded by a thin layer of cytoplasm with numerous lipid bodies.ConclusionThe study establishes a new host record, calanoid copepod B. similis for V. pelagica. The report is the first of its kind from the Indian waters and extends the biogeographic boundary of the apostome ciliate.

P-199 Effect of the area of oocyte perivitelline space on the fertilization and embryo development following intracytoplasmic sperm injection

Abstract Study question Does the area of oocyte perivitelline space have an effect on fertilization and embryo development following intracytoplasmic sperm injection? Summary answer The area of oocyte perivitelline space has not an effect on the fertilization but the embryo development following intracytoplasmic sperm injection. What is known already Oocyte perivitelline space has a lot of variation at intracytoplasmic sperm injection (ICSI). Some researchers reported that the characteristics of perivitelline space (large or small) affect embryo development, pregnancy, and implantation. However, these studies did not accurately calculate the area of perivitelline space. Therefore, little information is available on the effect of the area of oocyte perivitelline space on fertilization and embryo development following ICSI. The purpose of this study was to calculate and classify the area of oocyte perivitelline space and investigate the effect of the area of perivitelline space on fertilization and embryo development following ICSI. Study design, size, duration 1. We retrospectively investigated 634 mature oocytes that were conducted ICSI between January 2021 and December 2021. The area of each oocyte perivitelline space was defined from between the area of circle calculated from the inner layer of zona pellucida and cytoplasm and divided into 3 groups (-9%, 10-19%, 20%-). 2. We retrospectively calculated the diameter of an inner layer of zona pellucida and cytoplasm and compared it with the 3 groups (-9%, 10-19%, 20%-). Participants/materials, setting, methods 1. The fertilization, survival, good quality day-3 embryo, blastocyst, good quality blastocyst rates following ICSI were compared with the 3 groups (-9%, 10-19%, 20%-). 2. The average diameter of an inner layer of zona pellucida and cytoplasm of each oocyte for the 3 groups (-9%, 10-19%, 20%-) were compared. The data were analyzed by Fisher’s exact test, residual analysis, one-way ANOVA test, with Bonferroni correction as appropriate to determine the statistical differences among groups. Main results and the role of chance 1. The survival rates of perivitelline space -9%, 10-19%, 20%- groups were 100% (109/109), 96% (363/378), 94% (138/147), the fertilization rates were 89% (97/109), 88% (331/378), 86% (127/147), the good quality day-3 embryo rates were 56% (54/97), 70% (232/331), 70% (89/127) respectively. No significant difference was observed between these comparison items. The blastocyst rates of perivitelline space -9%, 10-19%, 20%- groups were 51% (47/92), 69% (222/321), 82% (93/114), the good quality blastocyst rates were 22% (20/92), 40% (129/321), 52% (59/114) respectively. The blastocyst and good quality blastocyst rates of perivitelline space -9% group showed significantly lower results. On the other hand, the blastocyst and good quality blastocyst rates of perivitelline space 20%- group showed significantly higher results. 2. The average diameter of an inner layer of zona pellucida of perivitelline space -9%, 10-19%, 20%- groups were 125 ± 4 µm, 129 ± 5 µm, 136 ± 6 µm, the average diameter of the cytoplasm of perivitelline space were 121 ± 4 µm, 119 ± 4 µm, 118 ± 4 µm respectively. Significant differences were observed in all pairs of groups of the average diameter of an inner layer of zona pellucida and cytoplasm. Limitations, reasons for caution The area of oocyte perivitelline space was calculated at only one plane. Wider implications of the findings Oocytes with narrow perivitelline space might have a wide region of adhesive between the cytoplasm surface and an inner layer of the zona pellucida which resulted in a smaller diameter of the zona pellucida and lower blastocyst rate by forming cytoplasmic fragments (Yumoto K et al. JARG. 2020 ;37(6):1349-1354.). Trial registration number Not Applicable

Gastrodermis ultrastructure of the different life stages of the polyopisthocotylean monogenean gill parasite Discocotyle sagittata

The polyopisthocotylean Discocotyle sagittata is a blood-feeding monogenean that infects the gill lamellae of rainbow trout, Oncorhynchus mykiss, and brown trout, Salmo trutta. The ultrastructure of their alimentary tract, at different stages of the life cycle, was previously unknown. Here, we show that the gastrodermis of the oncomiracidium, subadult, and adult D. sagittata follows the same structural organization as that of other blood-feeding polyopisthocotyleans, being composed of digestive cells alternating with a connecting syncytium. Digestive cells of the oncomiracidium are found in three developmental forms: undifferentiated, developing differentiated, and differentiated (presumably functioning) cells whereas those of adult and subadult are present in a single functioning state with variable size and content. The apical cytoplasm of adult digestive cells forms conical outgrowths, a feature which is absent in the oncomiracidium. The connecting syncytium of the oncomiracidium has no evidence of metabolic activity, while that of adult and subadult is metabolically active. The lamellae of the connecting syncytium of adults and subadults are more numerous and larger, and their terminal portions are expanded, compared with those of the oncomiracidium. Parallel, tubular, membranous structures are characteristic of the apical cytoplasm of the connecting syncytium of the oncomiracidium. Luminal lamella in the oncomiracidium, subadult, and adult form balloon-like structures enclosing some luminal contents, but those of the oncomiracidium are larger, bounded by nucleated cytoplasmic layer, and enclose more luminal contents. The possible functions of these structures and mechanism of digestion in both oncomiracidium and adult are discussed.

Functional ultrastructure and cytochemistry of vitellogenesis stages of Rohdella amazonica (Aspidogastrea, Aspidogastridae, Rohdellinae), a parasite of the Amazoninan banded puffer fish Colomesus psittacus

Vitellogenesis of Rohdella amazonica, parasites of the intestinal tract of a naturally infected banded puffer fish Colomesus psittacus, were studied by transmission electron microscopy (TEM) and TEM cytochemistry. Four vitellogenic stages were distinguished (I) stem cell stage of the gonial type (II) early differentiation stage, characterized by initiation of protein synthetic activity (III) an advanced differentiation and vitellocyte maturation stage, involving rapid accumulation of β-glycogen, large lipid droplets and smaller shell globules that fused progressively into large electron-dense shell globule clusters (IV) mature vitellocyte stage. The vitellogenesis process involves substantial increase in cell volume; extensive development of parallel cisternae of granular endoplasmic reticulum (GER) that become concentrically arranged in later stages and secrete shell globules; development of Golgi complexes engaged in the packaging of shell material; progressive accumulation, fusion and enlargement of unsaturated and saturated lipid droplets; formation of β-glycogen and α-glycogen and their accumulation into islands scattered among lipid droplets in maturing and mature vitellocytes; and finally during the advanced stage of maturation, rapid accumulation of large saturated lipid droplets α- and β-glycogen, and proteinaceous shell-globules and clusters in the peripheral cytoplasmic layer. Vitellogenesis in R. amazonica generally resembles that previously described for bothriocephalidean cestodes, but differs from that of other cestode orders. Positive cytochemical staining with periodic acid-thiocarbohydrazide-silver proteinate confirms β-glycogen and α-glycogen, around saturated lipid droplets of maturing and mature vitellocytes. These results from a basal trematode lead to a better understanding of the phylogenetic, functional, and developmental relationships among vitellogenesis patterns as they relate to egg formation, embryonic development, and life cycles among trematodes and cestodes.

Control of Listeria monocytogenes and Escherichia coli O157:H7 in enoki mushrooms (Flammulina velutipes) by combined treatments with organic acids, nisin, and ultrasound

This study was undertaken to evaluate the combined effect of organic acids, nisin, and ultrasound (US) on the reduction of Listeria monocytogenes and Escherichia coli O157:H7 in enoki mushrooms stored at 5 °C. Each bacterium was treated with 1–3% organic acids, 0.1% nisin, and 40 kHz US in enoki mushrooms individually or in combination at 25 °C for ≤30 min. Lactic acid (LA) was superior to malic acid, acetic acid, and nisin, decreasing L. monocytogenes and E. coli O157:H7 in enoki mushrooms by more than 3.00 log CFU/g at 25 °C for 30 min. US alone failed to inactivate L. monocytogenes and E. coli O157:H7 in enoki mushrooms. Particularly, L. monocytogenes and E. coli O157:H7 diminished to below the detection limits (<1.55 log CFU/g) in enoki mushrooms after dipping in 3% LA and 0.1% nisin incorporated with 40 kHz US at 25 °C for 30 min and remained constant incapable of growing in samples stored at 5 °C for 120 h. A transmission electron microscopy analysis presented collapse of peripheral cytoplasm layers with formation of local pores and breakage of cell wall structures in L. monocytogenes exposed to combined treatments with 3% LA, 0.1% nisin, and 40 kHz US. A combination of LA, nisin, and US exerted strong synergistic effects for controlling the mushroom carriage of L. monocytogenes and E. coli O157:H7.

Common differentially expressed genes and pathways correlating both coronary artery disease and atrial fibrillation.

Coronary artery disease (CAD) and atrial fibrillation (AF) share common risk factors, such as hypertension and diabetes. The patients with CAD often suffer concomitantly AF, but how two diseases interact with each other at cellular and molecular levels remain largely unknown. The present study aims to dissect the common differentially expressed genes (DEGs) that are concurrently associated with CAD and AF. Two datasets [GSE71226 for CAD) and GSE31821 for AF] were analyzed with GEO2R and Venn Diagram to identify the DEGs. Signaling pathways, gene enrichments, and protein-protein interactions (PPI) of the identified common DEGs were further analyzed with Kyoto Encyclopedia of Gene and Genome (KEGG), Database for Annotation, Visualization and Integrated Discovery (DAVID), and Search Toll for the Retrieval of Interacting Genes (STRING). 565 up- and 1367 down-regulated genes in GSE71226 and 293 up- and 68 down-regulated genes in GSE31821 were identified. Among those, 21 common DEGs were discovered from both datasets, which lead to the findings of 4 CAD and 21 AF pathways, 3 significant gene enrichments (intracellular cytoplasm, protein binding, and vascular labyrinthine layer), and 3 key proteins (membrane metallo-endopeptidase (MME), transferrin receptor 1 (TfR1), and Lysosome-associated membrane glycoprotein 1 (LAMP1)). Together, these data implied that these three proteins may play a central role in development of both CAD and AF.

Standing surface acoustic waves, and the mechanics of acoustic tweezer manipulation of eukaryotic cells

Manipulation by focused ultrasound is an emerging technology with much promise for non-contact handling of microscale objects. A particularly promising approach for achieving this with living cells involves incorporating standing surface acoustic waves (SSAWs) into a microfluidic device. SSAWs must be tuned to provide the necessary range of acoustic radiation force (ARF), but models enabling this tuning have neglected the mechanics of the cells themselves, treating cells as rigid or homogenous spheres, and have also neglected energy transfer from the substrate to the fluid at the Rayleigh angle. We therefore applied Mie scattering theory to develop a model of the ARF arising from a SSAW impacting an idealized eukaryotic cell in an inviscid fluid. The cell was treated as a three-layered body with a nucleus, cytoplasm, and cortical layer. Results showed strong dependence on cell structures and the Rayleigh angle that can be harnessed to develop novel applications for cell manipulation and sorting. ARF can be tuned using the new model to both push away and pull back a cell towards the sound source. The proposed analytical model provides a foundation for design of microfluidic systems that manipulate and sort cells based upon their mechanical properties.

Autophagy-mediated compartmental cytoplasmic deletion is essential for tobacco pollen germination and male fertility

ABSTRACT In plants, macroautophagy/autophagy has mainly been associated with stress-related processes but how it impacts normal physiological and developmental processes remains largely unexplored. Pollen germination is the critical first step toward fertilization in flowering plants. It is metabolically demanding and relies on high levels of cytoplasmic reorganization activities to support a dramatic morphological transformation that underlies the development of a pollen tube as the conduit to deliver sperm for fertilization. The role of autophagy in this process remains unclear. Here we provide evidence that pollen germination is accompanied by elevated autophagic activity and successful pollen tube emergence depends on autophagy-mediated cytoplasmic deletion. Genetic and cytological experiments demonstrate that inhibition of autophagy prevents pollen germination while induces the persistence of a layer of undegraded cytoplasm at the germination aperture. Together, these results unveil a novel compartmentalized autophagy. Furthermore, high-throughput comparative lipidomic analyses show that suppressed autophagy-induced inhibition of pollen germination is accompanied by altered profiles of stored and signaling lipids. Proteomic analyses reveal that autophagy likely exert its role in pollen germination via downstream mitochondria-related pathways. These findings reveal a critical role for autophagy in initiating pollen germination and provide evidences for compartmental cytoplasmic deletion being crucial for male fertility. Abbreviations: 3-MA: 3-methyladenine; ATG: autophagy-related gene; Cer: ceramide; CL: cardiolipin; Con A: concanamycin A; DAG: diradylglycerol; GO: gene ontology; HAG: hour after germination; LC-MS: liquid chromatography-mass spectrometry; MAG: min after germination; MDC: monodansylcadaverine; PE: phosphatidylethanolamine; PI: phosphatidylinositol; PLD: phospholipase D; PtdIns3K: phosphatidylinositol 3-kinase; RT-qPCR: quantitative real-time reverse transcription PCR; TAG: triradylglycerol; TEM: transmission electron microscopy; TMT: tandem mass tagging.

Ultrastructure of male Centrorhynchus globocaudatus (Acanthocephala) cement apparatus and function of cement gland secretion.

Cement glands are one of the most conspicuous and distinctive elements of taxonomic interest in male Acanthocephala. Cement glands vary in shape, number and arrangement in different classes of the taxon. The glands and their products have a fundamental role in the reproductive process. Light and electron microscopy were used to investigate the ultrastructure of the cement apparatus, which includes both cement glands and the cement reservoir, in mature males of Centrorhynchus globocaudatus (Zeder, 1800). Centrorhynchus globocaudatus is an enteric parasite of birds of prey, including Falco tinnunculus (Linnaeus, 1758) and Buteo buteo (Linnaeus, 1758) from the province of Ferrara (northern Italy). The four elongated cement glands of C. globocaudatus are situated posterior to the testes. Sections through the cement glands show each gland is surrounded by a fibrous envelope with an approximate thickness of 0.6 μm. Beneath this envelope is an outer cytoplasmic layer thickness ranging from 22 to 26 μm, which contains a number of nuclei with diameters variable from 20 to 22 μm. The cytoplasmic layer is filled with prominent free ribosomes and many mitochondria with lamellar cristae. Secretory granules, measuring from 1 to 1.3 μm in diameter, are formed within the cytoplasmic layer. The cytoplasmic layer surrounds the luminal area for storage of the cement material in each gland. Cement gland ducts arise from the gland and extend towards a common cement reservoir in close contact with the seminal vesicle and Saefftigen's pouch. Microtubules, large secretory granules and rest of undefined organelles were also observed within the cement reservoir.

Robin-type boundary conditions in transition from reaction-diffusion equations in 3D domains to equations in 2D domains

We consider a singular limit of diffusion equations in 3D domains of thickness converging to zero. In the 2D limit the resulting reaction-diffusion equation has a source term resulting from the Robin-type boundary conditions imposed on boundaries of the original 3D domain. The proposed approach can be applied to constructing approximate solutions of diffusion problems in thin planar, cylindrical, or spherical layers between two membranes. As an example we refer to the problem of activation of B lymphocytes, which typically have large nuclei and a thin cytoplasmic layer which can be considered as a spherical shell. For this example, assuming additionally axial symmetry we provide a rigorous convergence theorem in the language of semigroups of operators.

Seasonal expressions of luteinising hormone receptor, follicle-stimulating hormone receptor and prolactin receptor in the epididymis of the male wild ground squirrel (Spermophilus dauricus).

Luteinising hormone (LH), follicle-stimulating hormone (FSH) and prolactin (PRL) are pituitary-derived hormones and mediate their functions through LH receptor (LHR), FSH receptor (FSHR) and PRL receptor (PRLR) respectively. This study aimed to investigate the seasonal expression patterns of LHR, FSHR and PRLR in the epididymis of the male wild ground squirrel during the breeding and non-breeding seasons. Histologically, principal cells, basal cells, cilia and mature spermatozoa were found in the lumen of caput, corpus and cauda epididymidis in the breeding season, whereas in the non-breeding season, cilia and basal cells were rarely found and the epididymidal duct was devoid of spermatozoa. Immunohistochemical results showed that LHR, FSHR and PRLR were mainly present in the filamentous cytoplasm layer of epithelial cells of the caput, corpus and cauda epididymidis and FSHR and PRLR displayed stronger staining in the breeding season than in the non-breeding season. Furthermore, the mRNA and protein levels of FSHR and PRLR in all regions of epididymis as well as the levels of LHR in the caput and cauda epididymidis were higher during the breeding season. The protein levels of FSHR, LHR and PRLR were positively correlated with epididymal weight. Together, these results suggest that LHR, FSHR and PRLR may regulate epididymal functional changes in the male wild ground squirrel during its seasonal breeding cycle.

107 Insights from roe deer oocyte transcriptome across embryonic diapause

Embryonic developmental arrest, known as diapause, has been reported in more than 130 species. However, its mechanisms are still not completely understood. In the roe deer, the only known ungulate that exhibits this phenomenon, diapause lasts for approximately 5 months, starting after the rut period in mid-July to early August and ending with embryo elongation and implantation in December/January. Little is known regarding oocyte characteristics during this period. Here, we analysed the roe deer oocyte transcriptome as a model to understand diapause effects on oocyte features during embryonic developmental arrest and reactivation. During regular hunting, immature oocytes were obtained by ovary slicing from diapause and nondiapause stages, and classified according to morphological characteristics. Only oocytes with &amp;gt;2 layers of compact cumulus cells and cytoplasm from 30 hunted females were used for analyses. Immature oocytes were denuded and snap frozen. Additional oocytes were cultured in maturation medium for 20-24h. Matured oocytes with a present polar body were snap frozen. Two pools of 10 immature and mature oocytes for both diapause and nondiapause stages were included (at least 4 donors/pool). Oocyte pools were processed using the Smart-seq 2 single-cell protocol (Illumina Inc., San Diego, CA, USA) for full-length cDNA and library preparation. We performed RNA-seq on an Illumina sequencer. The obtained Fastq files were clipped and analysed with a locally installed version of the Galaxy platform. Sequences were mapped against the roe deer transcriptome (unpublished data) and annotated against human and bovine transcripts. Differentially expressed genes (DEG; false discovery rate &amp;lt;1%) were identified using EdgeR (https://bioconductor.org/packages/release/bioc/html/edgeR.html) in immature (IMM) and mature (M) oocytes from diapause (D) and nondiapause (ND) stages. Additionally, to evaluate the effects of maturation on oocyte transcript abundancy, DEG between IMM and M oocytes for D and ND were identified. Multidimensional scaling resulted in clustering according to oocyte types. Gene ontology terms for biological processes were assigned using ToppCluster tools. A total of 23066, 23022, 22438, and 22532 transcripts were detected for IMM D, M D, IMM ND and M ND oocytes, respectively. Using false discovery rate filtering, we found 333 and 288 DEG in immature and mature oocytes, respectively. Furthermore, maturation changed the oocyte expression profiles during D (2233 DEG) and ND periods (2589 DEG). Gene ontology classification revealed that most of the DEG in immature oocytes were involved in macromolecule catabolic process and oxidation-reduction process. For mature oocytes, DEG were mostly related to regulation of chromosome condensation and ER and Golgi vesicle-mediated transport. Additionally, the top 20 DEG from IMM D v. M D oocytes were related to mRNA splicing, mRNA metabolic process, and mRNA processing. These processes were not identified in the top 20 DEG from IMM ND v. M ND oocytes. These preliminary results suggest that oocyte transcriptome analysis could disclose new pathways implicated in oocyte competence and embryonic developmental arrest and activation stages.

Effect of nagilactone E on cell morphology and glucan biosynthesis in budding yeast Saccharomyces cerevisiae

Nagilactones are norditerpene dilactones isolated from the root bark of Podocarpus nagi. Although nagilactone E has been reported to show antifungal activities, its activity is weaker than that of antifungals on the market. Nagilactone E enhances the antifungal activity of phenylpropanoids such as anethole and isosafrole against nonpathogenic Saccharomyces cerevisiae and pathogenic Candida albicans. However, the detailed mechanisms underlying the antifungal activity of nagilactone E itself have not yet been elucidated. Therefore, we investigated the antifungal mechanisms of nagilactone E using S. cerevisiae. Although nagilactone E induced lethality in vegetatively growing cells, it did not affect cell viability in non-growing cells. Nagilactone E-induced morphological changes in the cells, such as inhomogeneous thickness of the glucan layer and leakage of cytoplasm. Furthermore, a dose-dependent decrease in the amount of newly synthesized (1, 3)-β-glucan was detected in the membrane fractions of the yeast incubated with nagilactone E. These results suggest that nagilactone E exhibits an antifungal activity against S. cerevisiae by depending on cell wall fragility via the inhibition of (1, 3)-β-glucan biosynthesis. Additionally, we confirmed nagilactone E-induced morphological changes of a human pathogenic fungus Aspergillus fumigatus. Therefore, nagilactone E is a potential antifungal drug candidate with fewer adverse effects.