First report of Caenorhabditis brenneri (Nematoda: Rhabditida) isolated from the cadaver of Philippinella moellendorffi (Stylommatophora: Ariophantidae), a terrestrial slug in the Philippines.

Gastropod-associated nematodes have been previously studied and documented worldwide, with some species forming host-specific association as obligate parasites of molluscs while others form intermediate and temporary association. Philippinella moellendorffi from Imelda, Zamboanga Sibugay, Philippines, were collected, washed and maintained in the laboratory until death. Cadavers were placed on nutrient agar to allow nematode proliferation. Nematode pure culture was obtained using one gravid female for propagation. Morphology and molecular analyses (18S ribosomal DNA (rDNA) and D2-D3 expansion segments of 28S rDNA) were employed as diagnostic tools in identifying the nematode species isolated from P. moellendorffi. The newly isolated nematode was identified as Caenorhabditis brenneri, thus designated as C. brenneri strain IZSP from the Philippines. This is the first record of C. brenneri isolated from the terrestrial slug P. moellendorffi.

Cerebellar Clustering and Functional Connectivity During Pain Processing.

The cerebellum has been traditionally considered a sensory-motor structure, but more recently has been related to other cognitive and affective functions. Previous research and meta-analytic studies suggested that it could be involved in pain processing. Our aim was to distinguish the functional networks subserved by the cerebellum during pain processing. We used functional magnetic resonance imaging (fMRI) on 12 subjects undergoing mechanical pain stimulation and resting state acquisition. For the analysis of data, we used fuzzy c-mean to cluster cerebellar activity of each participant during nociception. The mean time courses of the clusters were used as regressors in a general linear model (GLM) analysis to explore brain functional connectivity (FC) of the cerebellar clusters. We compared our results with the resting state FC of the same cluster and explored with meta-analysis the behavior profile of the FC networks. We identified three significant clusters: cluster V, involving the culmen and quadrangular lobules (vermis IV-V, hemispheres IV-V-VI); cluster VI, involving the posterior quadrangular lobule and superior semilunar lobule (hemisphere VI, crus 1, crus 2), and cluster VII, involving the inferior semilunar lobule (VIIb, crus1, crus 2). Cluster V was more connected during pain with sensory-motor areas, cluster VI with cognitive areas, and cluster VII with emotional areas. Our results indicate that during the application of mechanical punctate stimuli, the cerebellum is not only involved in sensory functions but also with areas typically associated with cognitive and affective functions. Cerebellum seems to be involved in various aspects of nociception, reflecting the multidimensionality of pain perception.

[The d.p.r. 37/97 and the thermal comfort of the surgeons according to the new version of UNI EN ISO 7730]. / Il d.p.r. 37/97 e il benessere termico dello staff operatorio alla luce della nuova versione dello standard UNI-EN-ISO 7730.

The Italian law, through a specific government decree (d.p.r. 37/97), recommends the minimal technological, structural and organising requirements that surgery rooms of public hospitals must assure. Nevertheless, thermal comfort is usually evaluated by the indices PMV and PPD (Fanger's indices). Microclimatic data, acquired inside 80 surgery rooms over 20 public hospitals located in South of Italy, have been reviewed taking into account the new version of the international standard UNI EN ISO 7730. This version enables to classify the workers in three categories, A, B and C each one related to a different value of the acceptable number of dissatisfied people PPD. Considering the particular task which surgeons and nurses are involved in, a percentage of dissatisfied < 6% appears more realistic and conservative respect to the 10% prescribed in the old version of the standard. So the calculations show that, in this case, less than 17% of the staff would considered the thermal conditions as comfortable. Furthermore, the range of the microclimatic parameters, as established by the low, is not sufficient to assure thermal comfort using the PMV or PPD.

Amalgam is a ligand for the transmembrane receptor neurotactin and is required for neurotactin-mediated cell adhesion and axon fasciculation in Drosophila.

Neurotactin (NRT), a member of the cholinesterase-homologous protein family, is a heterophilic cell adhesion molecule that is required for proper axon guidance during Drosophila development. In this study, we identify amalgam (AMA), a member of the immunoglobulin superfamily, as a ligand for the NRT receptor. Using transfected Schneider 2 cells and embryonic primary cultures, we demonstrate that AMA is a secreted protein. Furthermore, AMA is necessary for NRT-expressing cells both to aggregate with themselves and to associate with embryonic primary culture cells. Aggregation assays performed with truncated NRT molecules reveal that the integrity of the cholinesterase-like extracellular domain was not required either for AMA binding or for adhesion, with only amino acids 347-482 of the extracellular domain being necessary for both activities. Moreover, the NRT cytoplasmic domain is required for NRT-mediated adhesion, although not for AMA binding. Using an ama-deficient stock, we find that ama function is not essential for viability. Pupae deficient for ama do exhibit defasciculation defects of the ocellar nerves similar to those found in nrt mutants.

Formation of the peripheral nervous system during tail regeneration in urodele amphibians: ultrastructural and immunohistochemical studies of the origin of the cells.

In the regenerating newt tail, epimorphic regeneration--which recapitulates morphologically normal embryonic development--proceeds along a rostrocaudal differentiation gradient. Innervation of the new myomeres results from the spinal roots of segments rostral to the amputation plane and from ventral roots emerging from the lateroventral region of the regenerating spinal cord, in which motor neurons are differentiating. Electron microscopy and an indirect immunofluorescence study with anti-glial fibrillary acid protein (GFAP) confirm that the ventrolateral part of the regenerated ependymal tube gives rise to cells of the ventral root sheath and the spinal ganglia. Anti-GFAP and anti-neurofilament antibodies showed that ependymoglial cells and Schwann cells may play a role in neuronal pathfinding by helping guide and stabilize pioneering axons as they extend toward the myomeres. The carbohydrate epitope NC-1 is expressed in the spinal cord, in sheath cells of the spinal ganglia and in the non-myelin-forming Schwann cells of the peripheral nervous system. L1, a Ca++ independent neural cell adhesion molecule, was detected in the axonal compartments of the regenerating spinal cord, on immature and/or non-myelin-forming Schwann cells within the peripheral nervous system (PNS), and on nerve fibers within the regenerate. These immunohistochemical observations collectively support the hypothesis that Schwann cells already present in the blastema could be involved in organizing neural pathways.

Alterations in brain polyribosomal RNA translation and lymphocyte proliferation in prenatal ethanol-exposed rats.

The long-term effects of prenatal ethanol exposure on the properties of brain polysomes and the proliferative responses of lymphocytes to mitogenic stimulation in adult offspring were assessed. Female Sprague-Dawley rats either ingested the control or 6.6% ethanol-containing Lieber-DeCarli liquid diet during the 3rd trimester of pregnancy. Controls were age-matched and pair-fed. At 42 to 72 days of age, ethanol effects were evaluated on the (1) polysomal properties in the cerebral hemispheres, cerebellum, and hippocampal regions of the brain after translation in a messenger RNA (mRNA)-dependent rabbit reticulocyte lysate system and (2) immunologic functions of lymphocytes cultured from spleen cells by measuring their responses to mitogenic stimulation. Results showed long-term adverse effects of in utero ethanol exposure on the polysomal RNA translation in each of the three brain regions tested with free polysomal mRNAs affected more than the bound polysomal mRNAs. Of these, the hippocampal region appeared to sustain the most injurious effects. In addition, a suppression of the mitogen-induced lymphocyte proliferative responses were present under these conditions. The degree of suppression varied with the specific mitogen used. Data suggest that the ethanol effects on the CNS and lymphocyte proliferation are most possibly irreversible, and in the case of the CNS, a post-translational modification by ethanol is indicated. The reduced lymphocyte responses are suggestive of a possible interference by ethanol of the synthesis of interleukin-2 (IL-2) and/or a reduced binding of IL-2 with its receptor (IL-2 receptors).

Raising polyclonal antibodies using nitrocellulose-bound antigen.

Highly specific antibodies directed against minor proteins, present in small amounts in biological fluids, or against nonsoluble cytoplasmic or membraneous proteins, are often difficult to obtain. The main reasons for this are the small amounts of protein available after the various classical purification processes and the low purity of the proteins.

Raising polyclonal antibodies using nitrocellulose-bound antigen.

Highly specific antibodies directed against minor proteins, present in small amounts in biological fluids, or against nonsoluble cytoplasmic or membraneous proteins, are often difficult to obtain. The main reasons for this are the small amounts of protein available after the various classical purification processes and the low purity of the proteins.

A method for the production of highly specific polyclonal antibodies.

Two polyclonal antibodies directed against paramyosin and tropomyosin from Owenia fusiformis (a marine polychete annelid) were obtained using a new method of immunization. After purification by two-dimensional gel electrophoresis, proteins were transferred onto a nitrocellulose sheet using the Western blot technique. The proteins bound to their cellulose support were injected into rabbits without Freund's adjuvant and without solubilization of nitrocellulose with dimethyl sulfoxide. Highly specific polyclonal antibodies were generated.

Electrophoretic Comparison of Mitochondrial Polypeptides from Maize Lines Susceptible and Resistant to Helminthosporium maydis Race T.

The mitochondrial polypeptides from maize lines susceptible and resistant to Helminthosporium maydis were studied by single- and two-dimensional gel electrophoresis. Approximately 120 polypeptides were detected by two-dimensional gel electrophoresis. However, it was not possible to detect qualitative differences between the mitochondrial polypeptides of the two inbreds. These observations are discussed with reference to the putative mechanism of action of the pathotoxin.