Tubulin posttranslational modifications in in vitro matured prepubertal and adult ovine oocytes.

Microtubules (MTs), polymers of alpha/beta-tubulin heterodimers, are involved in crucial functions in eukaryotic cells. MTs physiology can be influenced by a variety of post-translational modifications (PTMs), including tyrosination, detyrosination, delta 2 modification, acetylation, polyglutamylation, polyglycylation. In mammalian oocytes, MTs are essential for meiosis, regulating the formation of meiotic spindle and chromosomes movements. Considering that the patterns of tubulin PTMs (tyrosination, detyrosination, acetylation, polyglutamylation and delta 2 modification) have not been investigated in ovine oocytes, this study has been designed to investigate their presence and quantification in in vitro matured (IVM) adult and prepubertal ovine oocytes. Oocytes from adult and lamb Sarda ewes, regularly slaughtered at the local abattoir, were in vitro matured, fixed, and processed by indirect immunofluorescence and confocal microscopy analyses at metaphase II stage. Our results revealed a well detectable signal for total, tyrosinated and acetylated α-tubulin in meiotic spindle of both sheep and lamb oocytes. On the other hand, no immunopositivity were appreciable for detyrosinated, polyglutamylated, and delta 2 tubulin in meiotic spindle of both sheep and lamb oocytes. As regard the tyrosinated and the acetylated α-tubulin PTMs, through the quantification of the fluorescence intensity, we did not find significant differences in their expression in meiotic spindle of sheep, while in lamb the acetylated tubulin levels were predominant in comparison with tyrosinated. Our results in addition to investigating for the first time the different tubulin PTMs in the spindle organization of ovine oocytes, showed a different microtubule pattern between adult and prepubertal oocytes. The microtubule cytoskeleton survey may thus suggest further cues to better understand skill-related problems in in the acquisition of oocyte competence.

Changes in renal hemodynamics of undernourished fetuses appear earlier than IUGR evidences.

The present study used a sheep model of intrauterine growth restriction, combining maternal undernutrition and twinning, to determine possible markers of early damage to the fetal kidney. The occurrence of early deviations in fetal hemodynamics which may be indicative of changes in blood perfusion was assessed by Doppler ultrasonography. A total of 24 sheep divided in two groups were fed with the same standard grain-based diet but fulfilling either their daily maintenance requirements for pregnancy (control group; n=12, six singleton and six twin pregnancies) or only the 50% of such quantity (food-restricted group; n=12; four singleton and eight twin pregnancies). All the fetuses were assessed by both B-mode and Doppler ultrasonography at Day 115 of pregnancy. Fetal blood supply was affected by maternal undernutrition, although there were still no evidences of brain-sparing excepting in fetuses at greatest challenge (twins in underfed pregnancies). However, there were early changes in the blood supply to the kidneys of underfed fetuses and underfed twins evidenced decreases in kidney size.

Glucogenic treatment creates an optimal metabolic milieu for the conception period in ewes.

This study determined the influence of a short-term glucogenic nutritional treatment on circulating concentrations of glucose, insulin, insulin-like growth factor 1 (IGF-1), nonesterified fatty acids (NEFA), and urea, and on their correspondent levels in follicular fluid (FF) collected 12 h after the end of the treatment. After estrous synchronization with intravaginal progestagen-impregnated sponges, 20 Sarda ewes were randomly allocated into two experimental groups (GLU and WAT) and, from day 7 to day 10 (day 0 = day of sponge removal), the GLU group was gavaged with a glycogenic mixture, whereas the WAT group was gavaged with water (control group). Follicular development was stimulated by FSH administration from day 8 to 10. At day 11, ovaries were collected and follicular fluid processed. Plasma changes were assessed from day 6 to 11. In GLU group, circulating concentration of glucose (P < 0.0001), insulin (P < 0.0001), and IGF-1 (P < 0.01) rose significantly, whereas NEFA and urea concentrations decreased (P < 0.0001), as compared with controls. In particular, in FF the higher glucose concentrations found in GLU ewes compared with controls (P < 0.0001) were not accompanied by any increase in insulin and IGF-1 concentrations. NEFA (P < 0.0001) and urea (P < 0.0001) were lower in FF of GLU than WAT group, although NEFA clearance in the ovary proved to be less efficient than at the systemic level. No significant difference between groups was found in FF concentrations of pregnancy-associated plasma protein A (a protease regulating the levels of free IGF-1 in follicles), glutathione, and in its total antioxidant capacity. These results suggest that glycogenic mixture administration creates a suitable follicular microenvironment for the conception period in dairy ewes.

Life-history traits of the long-nosed skate Dipturus oxyrinchus.

This work investigates life-history traits of the long-nosed skate Dipturus oxyrinchus, which is a common by-catch in Sardinian waters. The reproductive variables were analysed from 979 specimens sampled during scientific and commercial hauls. Females (10·4-117·5 cm total length, LT ) attained larger sizes than males (14·5-99·5 cm LT ). To evaluate age and growth, a sub-sample of 130 individuals (76 females and 54 males) were used. The age was estimated by annuli counts of sectioned vertebral centra. Four models were used for the length-at-age data: the von Bertalanffy, the exponential, the Gompertz and the logistic functions. According to the Akaike's information criterion, the Gompertz model seemed to provide the best fitting curve (L∞ mean ± s.e.: 127·55 ± 4·90 cm, k: 0·14 ± 0·09, IP: 3·97 ± 0·90 years). The oldest female and male were aged 17 (115·5 cm LT ) and 15 years (96·0 cm LT ), respectively. Lengths at maturity were 103·5 cm for females and 91·0 cm for males, corresponding to 90% of the maximum observed length in both sexes. The monthly distribution of maturity stages highlighted an extended reproductive cycle, with spawning females and active males being present almost throughout the year, as confirmed by the gonado-somatic index. Ovarian fecundity reached a maximum of 26 yolked follicles with a mean ± s.e. size of 19·7 ± 6·5 mm.

Diet and feeding behaviour of longnosed skate Dipturus oxyrinchus.

A total of 255 longnosed skate Dipturus oxyrinchus caught in Sardinian waters (central-western Mediterranean Sea), was analysed with respect to fish total length (LT ), season and depth, in order to provide information on diet and feeding behaviour. Specimens ranging from 93 to 1153 mm LT , were collected at depths between 121 and 671 m, during experimental trawl surveys carried out from 2005 to 2010. The diet comprised crustaceans [prey specific index of relative importance (%IPSRI ) = 72·69], teleosts (%IPSRI = 10·28) and molluscs (%IPSRI = 10·94). Levins' index (Bi ) showed a narrow niche breadth (Bi = 0·35). The mean ± s.e. trophic level (TL ) was 3·63 ± 0·50. The analysis showed major ontogenetic changes in the feeding behaviour. Early life stages were characterized by a benthic diet, which changed to benthopelagic during growth. Mysids, particularly Lophogaster typicus (%IPSRI = 34·51), were the main prey items of immature individuals, replaced by euphausiids, mainly Meganyctiphanes norvegica (%IPSRI = 13·19), in maturing fish. Crustaceans became less important in mature specimens, being replaced by molluscs (%IPSRI = 28·99) and teleosts (%IPSRI = 24·56). A concomitant increase of the TL was recorded (mean ± s.e. = 3·41 ± 0·44, 3·75 ± 0·54 and 4·28 ± 0·61 for immature, maturing and mature individuals). These feeding patterns ensured low levels of intraspecific competition. This study provides new information about the role that the D. oxyrinchus plays in the marine food chain and data now essential to formulate new and effective management plans for this species.

First record of Chilomycterus reticulatus (Osteichthyes: Diodontidae) in the Mediterranean Sea.

The spotfin burrfish Chilomycterus reticulatus (405 mm standard length) is recorded for the first time on the shoreline of the Mediterranean Sea. The adult specimen was found dried on a beach near the south Sardinian Sea (S. Antioco's Island). Photographs and morphometrics of the specimen are given.

Biosynthesis of the topaquinone cofactor in copper amine oxidases--evidence from model studies.

Copper amine oxidases utilize 2,4,5-trihydroxyphenylalanine quinone (topaquinone) as a cofactor in enzymatic catalysis. This cofactor is formed from a tyrosine residue through a self-catalytic mechanism with the participation of the copper ion at the active site. Although pathways have been postulated for topaquinone biogenesis, portions of this scheme are still unclear. We utilized 4-tert-butyl-derived models for the putative intermediates of topaquinone generation and studied the effect of Cu(II) and Zn(II) ions on each autoxidative step from dopa- to topaquinone-like compounds at physiological pH (7.4). Several polyvinyl-alcohol-based soluble resins bearing mono- and di-hydroxyphenolic moieties were also prepared, and their tendency to give hydroxyquinonic structures when incubated at alkaline pH values was investigated. Our results confirm (although indirectly) the formation of dopa and dopaquinone during topaquinone biosynthesis. Moreover, we collected evidence that, following the formation of dopa, the role of the active-site copper ion in topaquinone biogenesis would be limited to the catalysis of the two subsequent quinonization steps (i.e. from dopa to dopaquinone and from topa to topaquinone), thus disfavoring the possibility of a direct intervention of the metal ion in the hydroxylation of dopaquinone. In particular, Cu(II) was shown to influence deeply the autoxidation of 1,2,5-trihydroxy-4-tert-butylbenzene, used as model of topa, both increasing the reaction rate and changing its mechanism. The mechanistic implications of these findings for the biogenesis of topaquinone and its analogs at the active site of various amine oxidases are discussed.

[Granular cell tumor of the male breast. A case report]. / Il tumore a cellule granulari della mammella maschile. Descrizione di un caso clinico.

Granular cell tumor of the male breast is an extremely rare neoplasm. The case of a 64-year-old man with a periareolar lump in the upper medial quadrant of his left breast of recent onset is reported. At clinical examination, it was a poorly circumscribed, irregular, hard, mobile 1 cm mass. No axillary lymph nodes were palpable. The patient was submitted to ultrasound scan which showed a 1 cm periareolar solid hypoechoic lesion with an acoustic shadowing and subsequently to fine needle biopsy that revealed the presence of also atypical ductal hyperplasia. For this reason he was submitted to excision biopsy. The definitive histology was of granular cell tumor. The immunohistochemical investigations showed an intense positiveness for S-100 protein; the staining with PAS showed a diffuse positiveness. At 18 months after treatment the patient had no recurrence of disease. Granular cell tumor of the male breast is a neoplasm whose differential diagnosis with carcinoma is often possible only with definitive histology. For this reason, also in front of a mammary lump strongly suspicious for carcinoma, it's always necessary to think it can be this neoplasm and therefore to wait for the necessary histopathologic confirmation.

Antley-Bixler syndrome: case report and review of the literature.

The purpose of this report is to describe a patient with a pattern of malformations very similar to those described by Antley and Bixler. They include craniosynostosis, midface hypoplasia with proptosis, ear anomalies, choanal stenosis, long tapered fingers with a bulbous tip, elbow joint contracture due to radio-ulnar synostosis and talipes equinovarus. Necropsy revealed cardiac and renal malformations. The infant died a few days after birth, of respiratory failure. Unlike previously described cases the elbow joint contracture was due to radio-ulnar synostosis rather than radio-humeral synostosis, she did not have long bone fractures and the femora were not markedly bowed. Since the first report in 1975, at least 23 cases have been reported.

Intramyelinic edema in chronic inflammatory demyelinating polyneuropathy.

We describe a patient affected by chronic inflammatory demyelinating polyneuropathy (CIDP) whose sural nerve biopsy revealed a marked edema of the myelin sheath. Since this change has been previously described in human peripheral nerves only in CIDP patients, we suggest that intramyelinic edema may have a role in the pathogenesis of demyelinating neuropathies.

Acute axonal idiopathic polyneuropathy: a Guillain-Barré syndrome variant?

We report 7 cases of acute polyneuropathy fitting the NINDS diagnostic criteria for GBS. Electrophysiological study and sural nerve biopsy revealed a picture of axonal polyneuropathy, without changes suggesting demyelination. We discuss whether the acute idiopathic axonal neuropathy belongs to the GBS spectrum or represents a separate clinico-pathological entity.