Occurrence of multiple sexual chromosomes (XX/XY1Y2 and Z1Z1Z2Z2/Z1Z2W1W2) in catfishes of the genus Ancistrus (Siluriformes: Loricariidae) from the Amazon basin

Loricariid catfishes show a predominance of homomorphism in sex chromosomes, but cases of simple and multiple systems were also found. Here we describe two cases of multiple sex chromosome systems in loricariids from Brazilian Amazonia. Males of Ancistrus sp.1 "Balbina" have a modal number of 2n = 39 chromosomes, fundamental number (FN) of 78, and karyotypic formula of 27 m + 10 sm + 2 st; females have 2n = 38 chromosomes, FN = 76, and 26 m + 10 sm + 2 st. Ancistrus sp.2 "Barcelos" has 2n = 52 chromosomes for both sexes, FN = 80 for males and FN = 79 for females. Karyotypic formula is 12 m + 12 sm + 4 st + 24a for males and 11 m + 12 sm + 4st + 25a for females. The two species show different arrangements of constitutive heterochromatin blocks, which are coincident with NORs and absent in sex chromosomes. We suggest a XX/XY(1)Y(2) mechanism for Ancistrus sp.1 "Balbina", and a Z(1)Z(1)Z(2)Z(2)/Z(1)Z(2)W(1)W(2) mechanism for Ancistrus sp.2 "Barcelos". The XX/XY(1)Y(2) mechanism here reported is the second known occurrence of this type of multiple sex chromosomes for Loricariidae and the third for Neotropical fishes; the mechanism Z(1)Z(1)Z(2)Z(2)/Z(1)Z(2)W(1)W(2) represents the first record among fishes. The presence of different sex chromosome systems in Ancistrus indicates a probable independent origin and suggests that the differentiation of sex chromosomes is evolutionarily recent among species in this genus.     

Suppression of neurodegeneration and increased neurotransmission caused by expanded full-length huntingtin accumulating in the cytoplasm

Huntington's disease (HD) is a dominantly inherited neurodegenerative disorder caused by expansion of a translated CAG repeat in the N terminus of the huntingtin (htt) protein. Here we describe the generation and characterization of a full-length HD Drosophila model to reveal a previously unknown disease mechanism that occurs early in the course of pathogenesis, before expanded htt is imported into the nucleus in detectable amounts. We find that expanded full-length htt (128Qhtt(FL)) leads to behavioral, neurodegenerative, and electrophysiological phenotypes. These phenotypes are caused by a Ca2+-dependent increase in neurotransmitter release efficiency in 128Qhtt(FL) animals. Partial loss of function in synaptic transmission (syntaxin, Snap, Rop) and voltage-gated Ca2+ channel genes suppresses both the electrophysiological and the neurodegenerative phenotypes. Thus, our data indicate that increased neurotransmission is at the root of neuronal degeneration caused by expanded full-length htt during early stages of pathogenesis.     

Active metabolites from Dunalia spinosa resinous exudates

Dunalia spinosa, a plant used in folk medicine for toothaches, breathing problems and cleansing wounds, was found active as antimicrobial and antioxidant. A new (E)-aurone rutinoside (dunaurone) has been isolated from the aerial parts of the plant, and its structure was determined by spectroscopic means. Lupeol, beta-sitosterol, scopoletin, quercetin and withaferin A were also found. All the extracts exhibited strong antimicrobial activity while dunaurone showed only weak antimicrobial inhibition against Klebsiella pneumoniae; in addition it presented a significant free radical scavenging activity.     

ARAP1 regulates EGF receptor trafficking and signalling

The activation state of the EGF receptor (EGF-R) is tightly controlled in the cell so as to prevent excessive signalling, with the dangerous consequences that this would have on cell growth and proliferation. This control occurs at different levels, with a key level being the trafficking and degradation of the EGF-R itself. Multiple guanosine triphosphatases belonging to the Arf, Rab and Rho families and their accessory proteins have key roles in these processes. In this study, we have identified ARAP1, a multidomain protein with both Arf GTPase-activating protein (GAP) and Rho GAP activities, as a novel component of the machinery that controls the trafficking and signalling of the EGF-R. We show that ARAP1 localizes to multiple cell compartments, including the Golgi complex, as previously reported, and endosomal compartments, where it is enriched in the internal membranes of multivesicular bodies. ARAP1 distribution is controlled by its phosphorylation and by its interactions with the 3- and 4-phosphorylated phosphoinositides through its five PH domains. We provide evidence that ARAP1 controls the late steps of the endocytic trafficking of the EGF-R, with ARAP1 knockdown leading to EGF-R accumulation in a sorting/late endosomal compartment and to the inhibition of EGF-R degradation that is accompanied by prolonged signalling.     

Langerhans cell histiocytosis reveals a new IL-17A-dependent pathway of dendritic cell fusion

IL-17A is a T cell-specific cytokine that is involved in chronic inflammations, such as Mycobacterium infection, Crohn's disease, rheumatoid arthritis and multiple sclerosis. Mouse models have explained the molecular basis of IL-17A production and have shown that IL-17A has a positive effect not only on granuloma formation and neurodegeneration through unknown mechanisms, but also on bone resorption through Receptor activator of NF-kappaB ligand (RANKL) induction in osteoblasts. Langerhans cell histiocytosis (LCH) is a rare disease of unknown etiology, lacking an animal model, that cumulates symptoms that are found separately in various IL-17A-related diseases, such as aggressive chronic granuloma formation, bone resorption and soft tissue lesions with occasional neurodegeneration. We examined IL-17A in the context of LCH and found that there were high serum levels of IL-17A during active LCH and unexpected IL-17A synthesis by dendritic cells (DCs), the major cell type in LCH lesions. We also found an IL-17A-dependent pathway for DC fusion, which was highly potentiated by IFN-gamma and led to giant cells expressing three major tissue-destructive enzymes: tartrate resistant acidic phosphatase and matrix metalloproteinases 9 and 12. IFN-gamma expression has been previously documented in LCH and observed in IL-17A-related diseases. Notably, serum IL-17A-dependent fusion activity correlates with LCH activity. Thus, IL-17A and IL-17A-stimulated DCs represent targets that may have clinical value in the treatment of LCH and other IL-17A-related inflammatory disorders.     

Thyroid hormone effect in human hepatocytes

We have already demonstrated that a combined treatment of methimazole and an antioxidant mixture improved the condition of hyperthyroid patients both biochemically and clinically. Elevated thyroid hormone levels might trigger signs and symptoms of hyperthyroidism through the increase of free radicals. To study the direct effect of thyroid hormone on cellular markers of oxidative stress, we carried out in vitro assays in which 0.1-20.0 nM T3 (6.5-1300.0 ng/dl) doses were added to culture media of the human hepatocyte cell line Hep G2 for 1-24 h. T3 increased malondialdehyde (MDA) and intracellular oxidized glutathione (GSSG) levels; SOD activity was also higher with hormone treatment, whereas catalase and glutathione peroxidase activities showed no variation at different T3 doses and during all experimental times. When ascorbic acid was added to the culture, the MDA level decreased and SOD activity was increased. With higher doses of T3 (e.g. 200 nM), cell death occurred (69% of apoptotic cells). The increase in SOD activity was not enough to overcome the effect of T3 since MDA and GSSG remained high during a 24-h experiment. We showed a beneficial effect of ascorbic acid when cells were exposed to a T3 dose of 20 nM, a higher level of hormone than that achieved in hyperthyroidism.     

Sewage bacteriophage photoinactivation by cationic porphyrins: a study of charge effect.

Photodynamic therapy has been used to inactivate microorganisms through the use of targeted photosensitizers. Recently the inactivation of bacteria in residual waters has been reported, but nothing is known about photoinactivation of environmental bacteriophages, which are often used as indicators of human enteric viruses. In this study we tested the effect of six cationic porphyrin derivatives with two to four charges on the photoinactivation of a sewage bacteriophage. A phage suspension of 5 x 10(7) PFU mL(-1) was exposed to white light (40 W m(-2)), during 270 min, at three photosensitizer concentrations (0.5, 1.0 and 5.0 microM). Tetra- and tricationic porphyrins inactivated the T4-like sewage phage to the limits of detection, but dicationic porphyrins did not lead to a significant decrease in phage viability. At the highest photosensitizer concentration (5.0 microM), the phage was completely inactivated (>99.9999% of inactivation, reduction of 7.2 log) after 270 min by the tetracationic porphyrin. Two of the tricationic derivatives also led to phage inactivation to the limit of detection. The rate of bacteriophage photoinactivation and the efficiency of the photosensitizer appeared to vary with the charge and with the substituents in the meso-positions of the porphyrin macrocycle. Tetra- and tricationic porphyrins can, therefore, be used as a new method for inactivating sewage bacteriophages that are frequently used as human enteric virus indicators. The complete inactivation of viruses with low light intensity means that this methodology can be used even on cloudy days and during winter, opening the possibility to develop new technologies for wastewater treatment.     

Lack of "hemichannel" activity in insulin-producing cells

Connexins and pannexins have been implicated in the formation of "hemichannels," which may account for the uptake and release of membrane-impermeant molecules in single cells. The in vivo existence of "hemichannels" and their protein composition is still debated. Investigations on these matters are complicated by the lack of adequate negative controls. In search for such essential controls, the authors have investigated transformed (MIN6 line) and primary insulin-producing cells. Here, the authors report that these cells, which express Cx36 and pannexin 1, cannot be shown to display functional "hemichannels," as evaluated by (1) uptake of the membrane-impermeant tracer ethidium bromide, whether in the presence or absence of extracellular Ca(2+), following stimulation of P2X(7) receptors, and after exposure to hypotonic medium; and (2) lack of exocytosis-independent release of endogenous ATP. Moreover, electrophysiological recordings indicated the absence of carbenoxolone-sensitive pannexin 1 currents evoked by membrane potentials above +30 mV. Thus, insulin-producing cells are expected to provide a useful tool in the further characterization of hemichannel composition, properties, and physiological relevance.