The epistatic interaction between the dopamine D3 receptor and dysbindin-1 modulates higher-order cognitive functions in mice and humans.

The dopamine D2 and D3 receptors are implicated in schizophrenia and its pharmacological treatments. These receptors undergo intracellular trafficking processes that are modulated by dysbindin-1 (Dys). Indeed, Dys variants alter cognitive responses to antipsychotic drugs through D2-mediated mechanisms. However, the mechanism by which Dys might selectively interfere with the D3 receptor subtype is unknown. Here, we revealed an interaction between functional genetic variants altering Dys and D3. Specifically, both in patients with schizophrenia and in genetically modified mice, concomitant reduction in D3 and Dys functionality was associated with improved executive and working memory abilities. This D3/Dys interaction produced a D2/D3 imbalance favoring increased D2 signaling in the prefrontal cortex (PFC) but not in the striatum. No epistatic effects on the clinical positive and negative syndrome scale (PANSS) scores were evident, while only marginal effects on sensorimotor gating, locomotor functions, and social behavior were observed in mice. This genetic interaction between D3 and Dys suggests the D2/D3 imbalance in the PFC as a target for patient stratification and procognitive treatments in schizophrenia.

Porous poly (L-lactic acid) scaffolds are optimal substrates for internal colonization by A6 mesoangioblasts and immunocytochemical analyses.

In this study, mouse mesoangioblasts were seeded onto bidimensional matrices within three-dimensional porous scaffolds of poly (L-lactic acid) (PLLA), in the presence or absence of a type I collagen coating. The cells were observed under a scanning electron microscope and tested for their adhesion, survival and proliferation. Immunolocalization of heat shock protein (Hsp) 70, an abundant and ubiquitous intracellular protein in these cells, was also performed in sectioned cell-containing scaffolds under a confocal fluorescence microscope to determine if in situ analysis of intracellular constituents was feasible. The data show that PLLA films allow direct cell adhesion and represent an optimal support for cell growth, and that the internal surfaces of PLLA polymeric sponges can be colonized by mesoangioblasts, which can be submitted for in situ confocal microscopic analyses for possible monitoring of timedependent expression of differentiation markers.

The ecology of the parasite population in micro-mammals in the Italian peninsula and islands.

The importance of micro mammals from many points of view, mainly with an ecological approach was stressed. The study of the spatial-temporal distribution of parasites in their hosts may be carried out in several ways. Tests done in collaboration with the Parasitology Laboratory in the Faculty of Chemistry at the University of Barcelona, the NRCS and the Department of Ecological Studies in Cosenza, have contributed to an understanding of the Helminth communities as relating to several intrinsic variables of microteriofauna as well as extrinsic ones, particularly those concerning environment, climate and season. These comparisons were made using statistical means which compared the categorical and dichotomic variables which would highlight risk differences and their effects on the system. Quantitative dependent variables were also considered in relation to the aforementioned qualitative variables. One of the models studied is that of logistic regression, which estimates the function of regression, connecting the probability of the presence of Helminth as a dependent variable, with biological and ecological parameters (independent variables) such as: gender, age, season of capture, bioclimate, biotope and trapping section.

Stress response in mesoangioblast stem cells.

Stem cells are presumed to survive various stresses, since they are recruited to areas of tissue damage and regeneration, where inflammatory cytokines and cytotoxic cells may result in severe cell injury. We explored the ability of mesoangioblasts to respond to different cell stresses such as heat, heavy metals and osmotic stress, by analyzing heat shock protein (HSP)70 synthesis as a stress indicator. We found that the A6 mesoangioblast stem cells constitutively synthesize HSP70 in a heat shock transcription factor (HSF)-independent way. However, A6 respond to heat shock and cadmium treatment by synthesizing HSP70 over the constitutive expression and this synthesis is HSF1 dependent. The exposure of A6 to copper or to a hypertonic medium does neither induce HSP70 synthesis nor activation of HSF1, while a constitutive binding of constitutive heat shock element binding factor was found. Together, these data suggest that mesoangioblasts constitutively express HSP70 as an 'a priori' activation mechanism, while they maintain the ability to respond to stress stimuli.

Factors which influence sperm ability to fertilize.

Different factors influence animal sperm ability to fertilize. Some of them are reviewed here, sperm motility, block to polyspermy, chemioattraction, sperm competition for fertilization. Old and new data are reported, as for example the new notions on sperm motility derived from site directed mutagenesis in rodents, the new notions on the odour receptors in mammalian sperm attraction and new notions on sperm competition, which is variable in different species.

Epidemiological surveillance of bluetongue in Sicily.

The authors describe the status of bluetongue (BT) since 13 October 2000, when the first outbreak was reported in Sicily. The results of the epidemiological surveillance programme, based on sentinel animals distributed over the entire region, are also given. In Sicily, the incidence of the disease is relatively low compared to some other areas in the Mediterranean Basin. Seventy-five outbreaks of the disease were recorded in the first three epidemics (October 2000 to May 2003). Overall morbidity was 13.25%, mortality 5.36% and the case fatality rate 41.49%. The Province of Catania seems to have been the worst affected; the incidence rate in August 2002 was 0.8%. The monthly incidence rate was calculated for sentinel animals of which the estimated total was 3 654, distributed in 63 areas. It is important to underline that in the period under consideration, a total of 2 382 animals was examined. During the surveillance period, which extended from September 2001 to May 2003, the incidence of BT peaked in September 2002, at 5.91% -/+ 0.979. The cumulative incidence rate from September 2001 to August 2002 and September 2002 to March 2003 was 4.53% -/+ 0.76 and 20.03% -/+ 1.85, respectively. The circulation of BT virus serotypes 2, 4, 9 and 16 is described, as revealed by seroconversion in sentinel animals.

A constitutive 70 kDa heat-shock protein is localized on the fibres of spindles and asters at metaphase in an ATP-dependent manner: a new chaperone role is proposed.

In the present study, double immunofluorescence and immunoblot analysis have been used to show that centrosomes, isolated from Paracentrotus lividus sea urchin embryos at the first mitotic metaphase, contain the constitutive chaperone, heat-shock protein (HSP) 70. More specifically, we demonstrate that centrosomes contain only the HSP70-d isoform, which is one of the four isoforms identified in P. lividus. We also provide evidence that p34(cell division control kinase-2) and t complex polypeptide-1 (TCP-1) alpha, a subunit of the TCP-1 complex, are localized on the centrosomes. Furthermore, inhibition of TCP-1 in vivo, via microinjecting an anti-(TCP-1 alpha) antibody into P. lividus eggs before fertilization, either impaired mitosis or induced severe malformations in more than 50% of embryos. In addition, we have isolated the whole mitotic apparatus and shown that HSP70 localizes on the fibres of spindles and asters, and binds them in an ATP-dependent manner. These observations suggest that HSP70 has a chaperone role in assisting the TCP-1 complex in tubulin folding, when localized on centrosomes, and during the assembling and disassembling of the mitotic apparatus, when localized on the fibres of spindles and asters.

[Video laparoscopy in abdominal emergencies]. / La videolaparoscopia nelle urgenze addominali.

BACKGROUND: Personal experience about the use of video laparoscopy (VL) in abdominal emergencies is reported. DESIGN: retrospective evaluation of patients observed in the last years. SETTING: General Surgery I. Policlinico, University of Palermo. SUBJECTS: 61 VL have been performed: 30 acute appendicitis, 21 acute cholecystitis, 4 perforated peptic ulcer, 1 haemoperitoneum by haemorrhaged luteal corpus, 2 pelvic inflammatory disease (PID), 1 terminal ileitis, 1 choledochal perforation after ERCP and 1 bleeding after CVL. INTERVENTIONS: the following interventions have been performed: 22 VL appendectomy, 8 VL-assisted appendectomy, 21 VL cholecystectomy, 1 VL duodenal suture, 3 minilaparotomic duodenal suture, 2 prophylactic VL-assisted appendectomy, in 1 patient with terminal ileitis and in 1 PID, 1 VL partial ovarian resection. In the case with choledochal perforation during ERCP a traditional cholecystectomy was performed with an outer biliary drainage. In the patient with bleeding after CVL the spontaneous haemostasis seen during VL was confirmed by laparotomy performed to exclude baro-haemostasis and to prevent from legal motivation. The procedure was only diagnostic in 1 patient with PID. MAIN OUTCOME MEASURES: the diagnostic and therapeutic value, versus traditional surgery have been valued. RESULTS: VL is useful both for a correct diagnosis and to lead a laparotomy if necessary, allowing an adequate peritoneal exploration and toilet without large incisions; the operation is therefore, in any case, less invasive. CONCLUSIONS: In our experience the usefulness of VL is clear in the suspect of acute appendicitis, acute cholecystitis, perforated peptic ulcer, haemoperitoneum and when diagnosis is not sure and in all other situations in which correct preoperative diagnosis is impossible. So this procedure is useful to make easy a correct diagnosis and to surgical treatment.

Constitutive hsp70 is essential to mitosis during early cleavage of Paracentrotus lividus embryos: the blockage of constitutive hsp70 impairs mitosis.

Localization of constitutive hsp70 in eggs and early embryos of sea urchin Paracentrotus lividus is shown by means of in situ immunostaining. An accumulation of this protein is shown in the mitotic structures (asters, spindles and centrosomes). Microinjection of anti-hsp70 antibodies into eggs causes impairment of formation of mitotic structures and of cell division. This impairment goes from a complete mitotic block, to irregular mitotic apparatus formation with irregular cleavage, depending upon the antibody concentration. The localization of hsp70 after antibody microinjection is also described. Blockage of mitotic apparatus formation by nocodazole also blocks the concentration of hsp70 molecules observed in nontreated eggs. That the constitutive hsp70 plays a role in sea urchin mitosis is indicated.

Effect of the IMPase inhibitor L690,330 on sea urchin development.

A variety of concentrations of the IMPase inhibitor L690,330 were added to sea urchin embryos. Immediate arrest of development was obtained for concentrations from 7.5 mm on. Concentrations lower than 3.5 mm permitted gastrulation but inhibited skeletogenesis and disturbed elongation along the animal-vegetal axis. The latter results are similar to those obtained by counteracting lithium effect with myoinositol, which are suggested to be due to partial relief of IMPase inhibition.

Sea urchin HSF activity in vitro and in transgenic embryos.

Evidence is provided for the presence at the physiological temperature of 20 degrees C of a heat shock transcriptor factor, HSF, in the nuclei of P.lividus embryos. This HSF is able to specifically bind in vitro the heat shock element, HSE, of the promoter of the hsp70 gene i.v., as suggested by DNA-protein binding reactions and DNAse I protection assays. Upon heat-shock, at the temperature of 31 degrees C, its ability to bind the HSE units becomes much higher. The HSF activated by heat-shock drives in vivo the transcription of the beta-galactosidase reporter gene in transgenic sea urchin gastrulae. An ATF-like transcription factor, widely described in other organisms but not at all in sea urchins, is also present in the nuclear extracts and is able to bind the consensus individuated in the hsp70 i.v. gene promoter.

Histochemical analysis of the haemopoietic tissue in Spirographis spallanzanii.

Physical and cytochemical techniques have demonstrated that in Spirographis spallanzanii there is a highly pigmented tissue surrounding the ventral and circular vessels. Melanin and carotenoids are absent in the clubshaped cells, but there is evidence for the presence of lipids, haemopigments and Fe+++, probably in connection with haemopoietic processes. Tests on the ala-dehydrase activity in this tissue confirm this hypothesis. From these results it can be concluded that in Spirographis spallanzanii this tissue is the site of synthesis of respiratory pigment.