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Guido Macchiarelli

Publications and source records attributed to Guido Macchiarelli.

13 recordsLinked to original sources
PubMed2006

Mancozeb adversely affects meiotic spindle organization and fertilization in mouse oocytes.

In this study the effects of mancozeb, a widely used ethylenebisdithiocarbamate fungicide, on mouse oocyte meiotic maturation and fertilization were analyzed. Oocyte cumulus cell-complexes were matured in vitro with or without increasing concentrations of the fungicide (from 0.001 to 1 microg/ml) that, due to its different stability in organic solvents and in water, was resuspended either in dimethyl sulfoxide or in culture medium. Although, about 95% of oocytes reached the metaphase II stage; mancozeb-exposed oocytes showed a dose-dependent increase of alterations in spindle morphology, and this negative effect was more evident when the fungicide was resuspended in culture medium. Under the latter culture condition, oocytes matured in the presence of 0.1 and 1 microg/ml mancozeb showed a significant reduction also in the formation of male and female pronuclei. These results indicate that mancozeb can adversely affect mammalian reproductive performance, likely by perturbing microtubular organization during meiotic maturation.

Animals↗
PubMed2006

Morphological patterns of angiogenesis in ovarian follicle capillary networks. A scanning electron microscopy study of corrosion cast.

To describe the morphological characteristics of the ovarian follicle (F) capillary neoformation and regression, the angiogenic figures were studied by means of scanning electron microscopy of corrosion casts in developing and mature F of rabbit, pig, and cow. Developing F showed gradual neoformation of thecal capillaries characterized by budding and then sprouting, likely from preexisting interstitial vessels. Postcapillary venules frequently showed vasoconstriction rings (sphincters). Vasodilation followed capillary elongation. Mature F, in addition to vessel elongation and dilation, also presented infolding of dilated capillary walls, followed by capillary duplication and sinusoidalization. Periovulatory F mainly showed functional changes, such as capillary dilation, signs of iperpermeabilization, and ischemia, the latter being limited to the apical follicular area. Vessel regression was characterized by thinning of capillaries and presence of avascular areas within the atretic F wall at any stage. This study showed two main types of angiogenic patterns. (a) longitudinal elongation (in series, sprouting angiogenesis) characterizing the initial phase of F development and (b) parallel duplication (in parallel, infolding or intussusceptive angiogenesis), ending in capillary lateral replication or splitting, secondary to functional microvascular changes. Indirect evidence of the establishment of postcapillary resistances contributing to capillary remodeling, was also shown. It is concluded that the sequence of capillary neoformation in mammalian ovarian F occurs in six steps: (1) budding, (2) sprouting (and elongation), (3) dilation, (4) infolding (intussusception), (5) duplication (splitting and elongation), and (6) sinusoidalization. Capillary regression hits F at any stage and characterizes F atresia.

Animals↗
PubMed2006

Ultrastructural characteristics of human granulosa cells in a coculture system for in vitro fertilization.

The use of somatic cells for cocultures during in vitro fertilization (IVF) is currently finalized to obtain a higher number of healthy and viable embryos with a high potential of implantation. Among the different cell lines that can be used as feeder cells for cocultures, granulosa cells (GCs) are autologous cells, safe and easy to recover. The aim of the present study was to analyze the fine structure of human GCs used in a coculture system to evaluate, from a morphodynamic point of view, their role in supporting embryo development. GCs were collected during oocyte pick-up, 36 h after human chorionic gonadotropin administration, from patients undergoing IVF procedures, who had given their informed consent to be included in this protocol. After coculture, GCs were fixed and processed for light microscopy (LM) and transmission electron microscopy (TEM). By LM, GCs appeared as clusters of loosely packed cells, irregularly rounded or polyhedral in shape, varying in diameter from 18 to 25 microm. Mitotic cells, as well as regressing elements (with pyknotic nuclei or dense cytoplasm) and cell fragments were occasionally observed. By TEM, the plasma membrane was irregular due to the presence of cytoplasmic evaginations. Linear and annular gap junctions between neighboring GCs were found. GC nuclei, rounded and eccentrically located, contained finely dispersed chromatin, one (often two) prominent nucleoli and, infrequently, peripheral patches of heterochromatin. Numerous organelles populated the GC cytoplasm, among them, mitochondria were rod-shaped or elongated, usually provided with tubular-vesicular cristae but occasionally showing atypical, longitudinally oriented cristae. Membranes of smooth endoplasmic reticulum, Golgi stacks and vesicles, secretory-like granules, cisternae of rough endoplasmic reticulum (RER), free ribosomes and polysomes, lysosomal-like bodies, microfilaments, and lipid droplets were also seen in the GC cytoplasm. In most cells, RER was scarcely represented and numerous lipid droplets filled the perinuclear space. On the contrary, some GCs contained an abundant RER and rare lipid droplets scattered in the cytoplasm. In conclusion, our data demonstrated the presence, in a coculture system, of GCs provided with ultrastructural characteristics typical of healthy, metabolically active, mostly steroidogenic cells. Protein-synthetic cells have also been detected. These data, evaluated at the light of biochemical and clinical studies, sustain the capability of human GCs cocultures to positively affect early embryo development in vitro by the secretion of steroids and proteins, putative "embryotrophic" factors.

Cells, Cultured↗
PubMed2005

Mancozeb exposure in vivo impairs mouse oocyte fertilizability.

Mancozeb is known to alter reproductive performance in exposed animals, but its specific mechanism of action is still unclear. We investigated whether in female mice of the F1 generation, mancozeb could affect oocyte ability to undergo complete meiotic maturation and fertilization. Female mice were treated with 50 and 500 mg/kg of mancozeb (or vehicle in the controls) from gestational day 2 to postnatal day 20. Results demonstrated that only at the highest dose, mancozeb induced a significant decrease in the number of ovulated eggs. Moreover, at this dose mancozeb caused a significant decrease of fertilizability related to a reduction of the formation of male and female pronuclei.

Animals↗
PubMed2005

3-D ultrastructural distribution of collagen in human placental villi at term in relation to vascular tree.

In order to understand the 3-D distribution of collagen in relation to vascularization, chorionic villi of human placentae, belonging to normal pregnancies at term, were studied by scanning electron microscopy (SEM) after alkali maceration techniques, and by transmission electron microscopy (TEM). The villous tree appeared made of an uninterrupted structure of collagen fibres. The collagen fibres connected the chorionic villi axis with their basal plates and organised differently according to the various levels of villous branching. The collagen of stem villi showed copious fibres. The external fibres (facing the villous surface) were arranged mainly longitudinally. The central core of the villi (inner fibres) were arranged concentrically around the wall of the fetal vessels. Both external and internal fibres formed stratified lamellae or small parallel bundles. The inner core of stem villi showed small holes housing capillary spaces. Mature intermediate and terminal villi showed a scarce amount of collagen arranged in thin concentric layer within the villous core, surrounding numerous dilated capillary and sinusoid spaces.These observations demonstrated that the extracellular matrix of human chorionic villi is highly compartmentalised and shows a variable structural 3-D distribution depending on the branching level of the villous tree, such a distribution ensures the most favourable microenvironment for feto-maternal exchanges and it is likely able to provide a modulated support to the developing chorionic fetal vessels and trophoblastic layer as well.

Adult↗
PubMed2005

Surface morphology of the zona pellucida surrounding human blastocysts obtained after in vitro fertilization.

Human zona pellucida (ZP) is maintained up to the blastocyst stage prior to hatching. In in vitro fertilized (IVF) embryos, it eventually acts as a morphodynamic interface between the cultured embryo and its microenvironment. Ultrastructural data on the ZP of IVF blastocysts are scarce in humans. We employed correlated phase contrast microscopy (PCM) and scanning electron microscopy (SEM) to study retrospectively the ultrastructural morphology of the ZP outer surface of 20 IVF human blastocysts from 16 Japanese patients (28-44 years of age, average 36.7+/-4.2) with a history of infertility. Blastocysts were derived from conventional in vitro fertilization (cIVF) (n = 10) and from intracytoplasmic sperm injection (ICSI) (n = 10). Both cIVF and ICSI groups included "clear blastocysts" (n = 5) and "dark blastocysts" (n = 5). By PCM, the clear blastocysts exhibited a regular, round-shaped contour and consisted of clear and voluminous cells. By SEM, they displayed a spongy ZP with numerous fenestrations formed by networked filaments. By PCM, dark blastocysts appeared irregularly shaped and often collapsed, and comprised dark cells and debris. By SEM, their ZP were smooth with remnants of compact fenestrations. In conclusion, viable blastocysts presented a normal ZP outer surface ultrastructure, whereas unhealthy blastocysts showed an altered ZP outer surface, comparable to that of immature/atretic oocytes. Such alterations could reflect sub-optimal culture conditions and/or could be related to blastocyst degenerative processes. The blastocyst ZP surface ultrastructure was unaffected by the fertilization technique (cIVF or ICSI). These data suggest that blastocyst survival in vitro is related to ZP ultrastructure maintenance.

Adult↗
PubMed2004

Granulosa cell-oocyte interactions.

Throughout oogenesis the oocyte and follicle cells establish an intricate system of mutual interactions that ultimately lead to the acquisition of their respective competences. Paracrine factors released by both cell types are believed to stimulate formation of the primordial follicle and support the initial phases of follicle growth. At the same time, these processes are also dependent on gap junction communication between the germinal and somatic compartment. At later stages of follicle development, activities released by the oocyte induce the adjacent granulosa cells to express a specialized phenotype. In their turn, these cells crucially regulate the ability of the oocyte to progress through the meiotic process and acquire full developmental potential.

Cell Communication↗
PubMed2004

Influence of thyroid hormone on mouse preantral follicle development in vitro.

OBJECTIVE: To evaluate the role of 3,3',5-triiodothyronine (T(3)) on ovarian follicle development in vitro. DESIGN: Experimental study. SETTING: University reproductive biology unit. ANIMAL(S): CD1 mice (female). INTERVENTION(S): In vitro culture. MAIN OUTCOME MEASURE(S): Assessment of follicle and oocyte development following a 6-day culture with FSH (100 mU/mL), (Bu)(2)cAMP (0.5 mM), and T(3) (1-100 nM), alone or in combination. RESULT(S): A high percentage of in vitro grown (IVG) FSH-stimulated follicles formed an antral cavity (AC), while the addition of T(3) (100 nM) significantly reduced this response in a statistically significant manner. A statistically significant recovery of AC formation was obtained by limiting exposure to 4 days of culture. Formation of AC was induced by (Bu)(2)cAMP, which prevented T(3)-mediated suppression of AC formation. Under different conditions, high proportions of germinal vesicle-arrested IVG oocytes displayed condensed chromatin configuration. The capacity to undergo germinal vesicle breakdown (GVBD) was similar in all groups, and a statistically significant reduction in the percentage of oocytes with PB1 was recorded when T(3) was added to FSH or (Bu)(2)cAMP. This ability was partially recovered by removing T(3) on day 4 of culture. CONCLUSION(S): Exposure to high T(3) concentrations can impair preantral follicle development, but this effect can be partly reverted by restoring a physiological hormonal milieu before follicle commitment to antral development.

Animals↗
PubMed2003

Morphodynamics of the follicular-luteal complex during early ovarian development and reproductive life.

Female reproductive activity depends upon cyclic morphofunctional changes of the ovarian tissue during the female's fertile period, but the primum movens of an active gonadal rearrangement can be found from early phases of embryo development. To offer a basic account of the main steps of ovarian dynamics, we review the morphofunctional behavior of the follicular-luteal complex in an integrated study using light microscopy and transmission and scanning electron microscopy as well as through the use of numerous drawings. Particular emphasis is given to some reproductive aspects including (1) germ-somatic cell relationships and onset of folliculogenesis during early gonadal development; (2) follicular development and oocyte-follicle cell associations through adult folliculogenesis, finally leading to ovulation; (3) morphodynamics of corpus luteum formation, development, and regression, and (4) degenerative processes involving germ and somatic cells. The results reported, many of which originated in our laboratory, arise from some experiments on laboratory mammals but mostly from a large selection of human specimens. The data obtained are integrated and correlated with classic reports as well as with current views. Crucial biochemical, histophysiological, and clinical aspects are also emphasized.

Animals↗
PubMed2003

Distribution of collagen fibers in the aggregated lymphoid follicles of swine terminal ileum.

The arrangement of the collagen bundles was studied in the Peyer's patches of swine terminal ileum, by means of light microscopy (using silver-impregnation technique and picrosirius F3BA staining) and scanning electron microscopy (after NaOH-maceration). The lymphoid tissue forms a large and continuous patch along the antimesenteric border. The follicles are disposed mainly in the tela submucosa and sometimes they reached in the tunica mucosa surface (follicle/dome structures). Some follicles are located in the lamina propria of the tunica mucosa. Light microscopy showed black and brown-stained fibers, and yellow and red, and green-stained fibers, respectively by silver impregnation technique and picrosirius red staining, in the tela submucosa. In this tela, by scanning electron microscopy, the collagen fibers appeared as thick bundles forming a network of parallel layers. This network was denser in the interfollicular than in the follicular area, and formed a capsule surrounding the lymphoid follicles. Our results pointed out that a clear correspondence exists between the findings of currently used light microscopy techniques and the scanning electron microscopy after alkali-water maceration method. The arrangement of the collagen fibers in the antimesenteric border of the tela submucosa suggested a functional compartmentalization within the aggregated lymphoid follicles. This could facilitate the antigen-to-cell and cell-to-cell interaction during the immune response and thus create a suitable microenvironment for an active cell metabolism. The tunica mucosa showed a porous structure and its frequent gaps were likely the sites through which lymphocytes and other cells could freely migrate thus participating in the immunological activities of these structures.

Animals↗
PubMedDate not supplied

Hepatocyte ultrastructural aspects after preoperative biliary drainage in pancreatic cancer patients with cholestatic jaundice.

BACKGROUND: The usefulness of preoperative biliary drainage in long-standing obstructive jaundice remains controversial. This study was designed to assess microscopic and ultrastructural changes in the morphology of hepatocytes from patients with obstructive jaundice treated with biliary drainage for 14 days. PATIENTS AND METHODS: In 8 patients with jaundice due to pancreatic neoplasms we obtained two fine-needle liver biopsies: the first during a transhepatic cholangiographic examination before placing preoperative drainage and the second at surgery, on day 14 of drainage. Biopsy samples were examined by light and electron microscopy and correlated to serum liver functional tests recorded before and after biliary drainage. RESULTS: Pre-drainage biopsy presented diffuse morphological signs of congestion and cholestasis, lipid and bilirubin cytoplasm inclusions, loss of bile canaliculi microvilli and diffuse bile canaliculi dilatation. After-drainage biopsies presented reduction of bile canaliculi dilatation, partial restoring of bile canaliculi microvilli and persistence of diffuse hepatocyte structural and ultrastructural changes. Patients' laboratory values (bilirubin, alkaline phosphatase, SGGT, SGOT and SGPT), that were significantly impaired before drainage, returned almost to normal within two weeks after drainage. CONCLUSION: Preoperative biliary drainage in patients with long-standing obstructive jaundice has received wide yet controversial support due to a well established pathophysiological background. The present findings of scarce recovery of hepatocyte changes after 14 days' drainage seemingly question its appropriateness.

Aged↗