Glycopattern analysis and structure of the egg extra-cellular matrix in the Apennine yellow-bellied toad, Bombina pachypus (Anura: Bombinatoridae)

We studied the glycopatterns and ultrastructure of the extra-cellular matrix (ECM) of the egg of theApennine yellow-bellied toad Bombina pachypus, by light and electron microscopy in order to determine structure,chemical composition and function. Histochemical techniques in light microscopy included PAS and AlcianBlue pH 2.5 and 1.0, performed also after b-elimination. Lectin-binding was tested with nine lectins (AAA,ConA, DBA, HPA, LTA, PNA, SBA, UEA-I, WGA). An inner fertilization envelope (FE) and five jelly layers(J1–J5) were observed, differing in histochemical staining, lectin binding and ultrastructure. Most glycans wereO-linked, with many glucosamylated and fucosylated residues. The fertilization envelope presented a perivitellinespace and a fertilization layer, with mostly neutral glycans. The jelly layers consisted of fibers and granules,whose number and orientation differed between layers. Fibers were densely packed in J1 and J4 layers,whereas a looser arrangement was observed in the other layers. Jelly-layer glycans were mostly acidic and particularlyabundant in the J1 and J4 layers. In the J1, J2 and J5 layers, neutral, N-linked glycans were also observed.Mannosylated and/or glucosylated as well as galactosyl/galactosaminylated residues were more abundant in theouter layers. Many microorganisms were observed in the J5 layer. We believe that, apart from their functions inthe fertilization process, acidic and fucosylated glycans could act as a barrier against pathogen penetration

Glycopattern analysis and structure of the egg extra-cellular matrix in the Apennine yellow-bellied toad, <i>Bombina pachypus (Anura: Bombinatoridae)</i>

We studied the glycopatterns and ultrastructure of the extra-cellular matrix (ECM) of the egg of the Apennine yellow-bellied toad <i>Bombina pachypus</i>, by light and electron microscopy in order to determine structure, chemical composition and function. Histochemical techniques in light microscopy included PAS and Alcian Blue pH 2.5 and 1.0, performed also after <i>b</i>-elimination. Lectin-binding was tested with nine lectins (AAA, ConA, DBA, HPA, LTA, PNA, SBA, UEA-I, WGA). An inner fertilization envelope (FE) and five jelly layers (J1–J5) were observed, differing in histochemical staining, lectin binding and ultrastructure. Most glycans were O-linked, with many glucosamylated and fucosylated residues. The fertilization envelope presented a perivitelline space and a fertilization layer, with mostly neutral glycans. The jelly layers consisted of fibers and granules, whose number and orientation differed between layers. Fibers were densely packed in J<sub>1</sub> and J<sub>4</sub> layers, whereas a looser arrangement was observed in the other layers. Jelly-layer glycans were mostly acidic and particularly abundant in the J<sub>1</sub> and J<sub>4</sub> layers. In the J<sub>1</sub>, J<sub>2</sub> and J<sub>5</sub> layers, neutral, N-linked glycans were also observed. Mannosylated and/or glucosylated as well as galactosyl/galactosaminylated residues were more abundant in the outer layers. Many microorganisms were observed in the J<sub>5</sub> layer. We believe that, apart from their functions in the fertilization process, acidic and fucosylated glycans could act as a barrier against pathogen penetration. (Folia Histochemica et Cytobiologica 2011; Vol. 49, No. 2, pp. 306–316

How temperature affects equine semen: refrigeration versus cryopreservation. A simple method to select high quality spermatozoa

Cooled and frozen equine semen shows a reduction in fertility, compared to fresh one. In this study, cooled and frozen-thawed equine spermatozoa were compared and analyzed for plasma and acrosomal membrane integrity and mitochondrial membrane potential, combining three fluorescent probes: H258, CTC, JC-1 with a micro-spectrofluorimetric analysis (Quanticell equipped with a digital system for color images acquisition). Total and progressive motility, average path velocity (VAP), straight-line velocity (VSL), curvilinear velocity (VCL) and amplitude of lateral head displacement (ALH) were measured by CASA system. We employed an innovative approach to study the reproductive potential of the male gamete subjected to cooling protocols for semen storage. In fact, we evaluated the modifications of equine sperm physiology induced by temperature during cooling and freezing treatments looking at the modifications of different functional sperm characteristics by a simultaneous analysis of different sperm markers with the aim of selecting those that are the most efficient signs for sperm fertility. We identified the mitochondrial membrane potential because it provides useful information on equine sperm quality strictly correlated with fertility. We consider it a useful marker for sperm fertility to be used as a guide to select high-quality semen to be employed in equine breeding farmers

Sars-CoV-2 isolation from a 10-day-old newborn in Italy: A case report

This report describes the evolution of COVID-19 in a 10 day-old-baby. The mother developed the disease immediately after childbirth and therefore a vertical transmission can be excluded. The isolation of the virus in cell culture with a cytopathic effect already visible after 48 h, indicates that the viral load of the newborn was quite high, but not serious course of the disease was observed. This paper wants to highlight the possible role of newborns and children in the spread of the disease