Diversification in gravel beaches: A radiation of interstitial clingfish (Gouania, Gobiesocidae) in the Mediterranean Sea.

We report a hitherto unknown radiation within the clingfishes (Gobiesocidae), discovered in one of the best-studied marine biomes, the Mediterranean Sea. The monotypic genus Gouania is a Mediterranean endemic inhabiting the interstices of gravel beaches. Using geometric morphometric analyses, we identified two distinct morphotypes (characterized by a slender and a stout body shape, respectively) among Gouania willdenowi sampled from the three major Mediterranean basins (Eastern, Western and Adriatic). Slender and stout G. willdenowi occurred sympatrically in the Adriatic and the Eastern Mediterranean basins. Only the stout morphotype was found in the Western Mediterranean. Morphotypes were further distinguished by relative eye size and number of vertebrae. Based on mitochondrial and nuclear gene sequences, five highly divergent clades (COI-based K2P distances of 8 to 15%) were identified. The clades represented, respectively, stout Adriatic, slender Adriatic, stout Eastern, slender Eastern and stout Western Mediterranean Gouania. This suggests that the genus Gouania comprises at least five different species. Phylogenetic relationships among clades furthermore imply repeated evolution of convergent morphotypes. The onset of the Gouania radiation was dated to 3.23 (95% HPD 2.08-5.90) mya, and may therefore have overlapped with or followed the Messinian salinity crisis.

Medullary thyroid carcinoma: autologous tumor cell lines for dendritic cell vaccination.

BACKGROUND: Medullary thyroid carcinoma (MTC) is a calcitonin-producing tumor of the parafollicular C-cells, accounting for 5-10% of all thyroid tumors. To date, the only effective treatment is the early and total surgical removal of all neoplastic tissue. As the prognosis of patients with advanced MTC, unresectable or distant metastases is poor, and chemotherapy or irradiation is of no significant value, alternative strategies have been sought. MATERIALS AND METHODS: A promising treatment approach for human MTC, that has already been introduced at our facility, is based on vaccination with autologous dendritic cells (DCs). Strong evidence that vaccination with autologous tumor lysate-pulsed DCs induces a specific immune response in vivo has been provided. However, the therapeutic success of this approach is sometimes critically limited by the small amount of tumor material available, especially from patients operated at an early tumor stage. Thus, it would be to the best advantage to have sufficient amounts of autologous tumor cells available for DC pulsing. RESULTS: A method to generate viable autologous tumor cell cultures from a variety of MTC tissue samples, even when the sample size is small, has been successfully established. These cell lines maintain their neuroendocrine phenotype. In addition, it can be shown that these cells also display the biological features of neuroendocrine tumor cells at the molecular level. CONCLUSION: The unlimited availability of these MTC cell lines makes it possible to specify cancerogenesis of MTC. In addition, the availability of sufficient amounts of tumor lysate from these cell lines offers the advantage of prolonged immunotherapy. Finally, these cell lines could be elegantly used as read-out system to monitor the in vivo immune response during immunotherapy with DC cell-based vaccination in patients suffering from MTC.

Galanin in the brain: chemoarchitectonics and brain cartography--a historical review.

We present a review of galanin in the brain from a historical perspective of the development of "chemoarchitectonics" and "brain cartography" accomplished in the Histopharmacology Section at the National Institutes of Health. It was the mapping of potential brain neuroregulators that served as a springboard of ideas from which behavioral studies emanate. The integration of the known localization of neurotransmitter/neuromodulatory nerves ("chemoarchitectonic maps") and receptor binding sites with biochemical data derived from brain micropunches coupled with behavioral analysis at the level of discrete brain allows one to define the anatomical circuits which support behavioral changes and which ultimately will improve our understanding of mental disorders.

Choline acetyltransferase, glutamic acid decarboxylase and somatostatin in the kainic acid model for chronic temporal lobe epilepsy.

The aim of the study was to investigate neurochemical changes in a kainic acid (KA; 10 mg/kg, s.c.)-induced spontaneous recurrent seizure model of epilepsy, 6 months after the initial KA-induced seizures. The neuronal markers of cholinergic and gamma-aminobutyric acid (GABA)ergic systems, i.e. choline acetyltransferase (ChAT) and glutamic acid decarboxylase (GAD) activities, and a marker for neuropeptide, i.e. level of somatostatin, have been investigated. The brain regions investigated were the hippocampus, amygdala/piriform cortex, caudate nucleus, substantia nigra and the frontal, parietal, temporal and occipital cortices. Six months after KA injection, reduced ChAT activity was observed in the amygdala/piriform cortex (47% of control; p<0.001), increased ChAT activity in the hippocampus (119% of control; p<0.01) and normal ChAT activity in the other brain regions. The activity of GAD was significantly increased in all analysed cortical regions (between 146 and 171% of control), in the caudate nucleus (144% of control; p<0.01) and in the substantia nigra (126% of control; p<0.01), whereas in the amygdala/piriform cortex, the GAD activity was moderately lowered. The somatostatin level was significantly increased in all cortical regions (between 162 and 221% of control) as well as in the hippocampus (119% of control), but reduced in the amygdala/piriform cortex (45% of control; p<0.01). Six months after KA injection, the somatostatin:GAD ratio was lowered in the amygdala/piriform cortex (49% of control) and in the caudate nucleus (41% of control), whereas it was normal in the hippocampus and moderately increased in the cortical brain regions. A positive correlation was found between seizure severity and the reduction of both ChAT activities and somatostatin levels in the amygdala/piriform cortex. The results show a specific pattern of changes for cholinergic, GABAergic and somatostatinergic activities in the chronic KA model for epilepsy. The revealed data suggest a functional role for them in the new network that follows spontaneous repetitive seizures.

Physiological leukocytosis during pregnancy is associated with changes in glucose transporter expression of maternal peripheral blood granulocytes and monocytes.

PROBLEM: The scarce data on glucose transporter expression of leukocytes are contradictory and nothing is known about changes accompanying physiological leukocytosis during pregnancy, which imposes acute metabolic demands on the cells. METHOD OF STUDY: Cytospin preparations of intravascular leukocytes were searched immunocytochemically for the high affinity glucose transporters GLUT1, 3 and 4. Pregnancy-associated quantitative changes in transporter expression were assessed by flow cytometry. RESULTS: Granulocytes and monocytes stained for GLUT1, 3 and 4. Major changes in cell surface transporter expression during pregnancy were a 36% (P < 0.05) down-regulation of granulocyte GLUT1 at term, and an increase in monocyte GLUT3 levels to 137% (P < 0.05), paralleled by a 24% (P < 0.05) decrease in GLUT4 content in second trimester. Apart from a minor subpopulation, lymphocytes were negative for these carriers. CONCLUSION: GLUT1, 3 and 4 are abundantly expressed in granulocytes and monocytes. The particular isoforms are differentially regulated during pregnancy, suggesting an individual functional significance.

In vitro models of brain ischemia: the peptidergic drug cerebrolysin protects cultured chick cortical neurons from cell death.

Glutamate (1 mM), iodoacetate (0.01 mM) and ionomycin (0.25 micro M) are reported to induce several characteristics of ischemia and neuronal degeneration in vitro, e.g. glutamate and ionomycin lesion result in a disturbance of Ca(2+) homeostasis, iodoacetate impairment leads to an inhibition of energy metabolism, suppression of protein synthesis and generation of oxygen free radicals. In this study these three lesion models were used to investigate the effects of the nootropic drug Cerebrolysin (Cere) on the survival of cortical neurons in culture and on the occurrence of apoptosis. The viability of the cells was evaluated with the colorimetric MTT-reduction assay. Apoptosis was detected with Bisbenzimide (Hoechst:33258), a fluorescent DNA stain. Administration of Cere resulted in dose dependent neuroprotection independent from the kind of lesion. In the glutamate model the drug almost doubled neuronal viability compared to lesioned controls. After acute glutamate exposure Cere reduced the number of apoptotic cells significantly. In spite of the protective efficacy after cytotoxic hypoxia induced by iodoacetate, the drug significantly increased the number of apoptotic neurons, indicating a shift from necrosis to apoptosis. In contrast to previous studies investigating acute ionomycin lesions, the chronic Ca(2+)-overload used here did not increase the abundance of apoptosis compared to the unlesioned control. Summarizing the findings it can be suggested that Cere is able to stabilize Ca(2+) homeostasis, to protect protein synthesis and to counteract neuronal death in different in vitro medels of ischemia.