Development changes in calla lily plants due to salt stress

Soil salinity is mainly caused by excessive use of fertilizers and the use of poor quality water, and adversely affected crop growth especially when grown in protected environments. Soil salinity causes salt stress in plants, which inhibits plant growth, leading to morphological, metabolic and physiological changes. Though it is a major problem occurs more frequently, there is less information on the behavior of calla lily (Zantedeschia aethiopica) under these conditions, and most studies are conducted with other species of the genus Zantesdeschia. Therefore, this study aimed to evaluate ecophysiological, biochemical and anatomical growth responses of calla lily plants to salt stress. Rhizomes were grown in trays containing coconut fiber as a substrate and treated with 0, 25, 50, 75 and 100 mM NaCl to induce stress. A decrease in plant height was observed, as well as in the number of tillers and leaves, main root length, fresh and dry matter of the shoot and root system. A reduction in photosynthetic rate, stomatal conductance and transpiration rate was observed at 60 days. However, after 90 days, the photosynthetic rate was unchanged, with increased stomatal conductance and transpiration rate for plants exposed to 75 mM NaCl. Salt stress caused a higher accumulation of carbohydrates in shoots and roots. Thus, high concentrations of NaCl affect the development of calla lily, indicating that this species is susceptible to salt stress.     

Locomotor Activity Rhythm in Free-ranging Common Marmosets (Callithrix jacchus)

Locomotor activity rhythm in the common marmoset has been well described and characterized in captivity, but there is no information about rhythmic aspects of this behavior in free-ranging conditions. To investigate if the locomotor activity rhythm in free-ranging common marmosets is similar to that found in captive common marmosets, we observed reproductive animals from two groups that inhabited at the Experimental Forestry Station, in northeastern Brazil (6°5'S, 35°12'W). From December 1992 to October 1993 we recorded the frequency of locomotor activity by focal animal sampling. The records of locomotor activity were submitted to spectral analysis and to the Cosinor test. Common marmosets showed an average duration of the active phase of 11:37 h ± 13.8 min. Similar to recorded in captive common marmosets, circadian rhythmicity was found with peaks in the morning, and in the afternoon, characterizing a bimodal pattern with peaks separated by 8 hours. The presence of a rhythmic pattern in the locomotor activity in free-ranging and captive common marmosets suggests that this pattern is not determined by the environmental conditions in captivity, but it has a endogenous component that play an important role for adaptation of this species to different conditions.     

Isolation of polymorphic microsatellite markers for the demersal fish Helicolenus dactylopterus (Dela Roche 1809)

Six microsatellite loci were identified for the demersal deep‐sea fish Helicolenus dactylopterus. All loci were highly polymorphic (5–21 alleles per locus). Observed heterozygosities were from 0.378 to 0.868, while the expected heterozygosities ranged from 0.529 to 0.925. Multiplex PCR reactions were optimized. Microsatellite markers were developed for analysis of genetic structure including identification of stocks and migration patterns. The resulting data will be used to help in the establishment of scientifically based fisheries management for this species. Departures from the expected Hardy–Weinberg equilibrium were observed in three loci, and are likely to be a consequence of population structuring across the Azorean islands.     

The Vesicular Acetylcholine Transporter Is Required for Neuromuscular Development and Function

The vesicular acetylcholine (ACh) transporter (VAChT) mediates ACh storage by synaptic vesicles. However, the VAChT-independent release of ACh is believed to be important during development. Here we generated VAChT knockout mice and tested the physiological relevance of the VAChT-independent release of ACh. Homozygous VAChT knockout mice died shortly after birth, indicating that VAChT-mediated storage of ACh is essential for life. Indeed, synaptosomes obtained from brains of homozygous knockouts were incapable of releasing ACh in response to depolarization. Surprisingly, electrophysiological recordings at the skeletal-neuromuscular junction show that VAChT knockout mice present spontaneous miniature end-plate potentials with reduced amplitude and frequency, which are likely the result of a passive transport of ACh into synaptic vesicles. Interestingly, VAChT knockouts exhibit substantial increases in amounts of choline acetyltransferase, high-affinity choline transporter, and ACh. However, the development of the neuromuscular junction in these mice is severely affected. Mutant VAChT mice show increases in motoneuron and nerve terminal numbers. End plates are large, nerves exhibit abnormal sprouting, and muscle is necrotic. The abnormalities are similar to those of mice that cannot synthesize ACh due to a lack of choline acetyltransferase. Our results indicate that VAChT is essential to the normal development of motor neurons and the release of ACh.Cholinergic neurotransmission has key functions in life, as it regulates several central and peripheral nervous system outputs. Acetylcholine (ACh) is synthesized in the cytoplasm by the enzyme choline acetyltransferase (ChAT) (16). Choline supplied by the high-affinity choline transporter (CHT1) is required to maintain ACh synthesis (52). A lack of ChAT (4, 35) or the high-affinity choline transporter (21) in genetically modified mice is incompatible with life. ACh plays an important role in wiring the neuromuscular junction (NMJ) during development (38, 43). Embryonic synthesis of ACh is fundamental for the development of proper nerve-muscle patterning at the mammalian NMJ, as ChAT-null mice present aberrant nicotinic ACh receptor (nAChR) localization and increased motoneuron (MN) survival, axonal sprouting, and branching (4, 35).The vesicular ACh transporter (VAChT) exchanges cytoplasmic ACh for two vesicular protons (37, 41). Previously reported electrophysiological studies showed that quantal size is decreased by vesamicol, an inhibitor of VAChT, but only in nerve terminals that have been electrically stimulated (19, 59, 60, 63). VAChT overexpression in developing Xenopus MNs increases both the size and frequency of miniature-end-plate currents (54). In Caenorhabditis elegans, mutations in VAChT affect behavior (65). Moreover, a decrease in VAChT expression has functional consequences for mammals, as mutant mice with a 70% reduction in the expression levels of this transporter (VAChT knockdown [KD^HOM] mice) are myasthenic and have cognitive deficits (47). Hence, vesicular transport activity is rate limiting for neurotransmission “in vivo” (18, 47).Exocytosis of synaptic vesicle contents is the predominant mechanism for the regulated secretion of neurotransmitters (55). However, alternative mechanisms of secretion have been proposed (20, 56, 61). Quantal ACh release, comparable to that seen in developing nerve terminals, has been detected in myocytes and fibroblasts in culture, which presumably do not express VAChT (14, 24). More recently, it was found that the correct targeting of Drosophila photoreceptor axons is disrupted in flies with null mutations in ChAT (64). Remarkably, the inactivation of VAChT did not produce the same result (64). The result suggests that the release of ACh during development is not dependent on VAChT, perhaps because it is nonvesicular or because vesicular storage can occur without VAChT.To test if the VAChT-independent secretion of ACh has any physiological role in the mammalian nervous system, we generated a mouse line in which the VAChT gene is deleted. These mice lack the stimulated release of ACh from synaptosomes, die after birth, and show several alterations in neuromuscular wiring consistent with a severe decrease in the cholinergic input to muscles during development. These experiments indicate that VAChT has an important role in maintaining activity-dependent ACh release that supports life and the correct patterning of innervation at the NMJ.     

Oxidative Stress in Cerebrospinal Fluid of Patients with Aseptic and Bacterial Meningitis

This study aimed to determine whether patients with aseptic and bacterial meningitis presented alterations in oxidative stress parameters of cerebrospinal fluid (CSF). A total of 30 patients were used in the research. The CSF oxidative stress status has been evaluated through many parameters, such as lipid peroxidation through thiobarbituric acid reactive substances (TBARS) and antioxidant defense systems such as superoxide dismutase (SOD), glutathione S-transferase (GST), reduced glutathione (GSH) and ascorbic acid. TBARS levels, SOD and GST activity increase in aseptic meningitis and in bacterial meningitis. The ascorbic acid concentration increased significantly in patients with both meningitis types. The reduced glutathione levels were reduced in CSF of patients with aseptic and bacterial meningitis. In present study we may conclude that oxidative stress contributes at least in part to the severe neurological dysfunction found in meningitis.     

A VANADIUM BROMOPEROXIDASE CATALYZES THE FORMATION OF HIGH‐MOLECULAR‐WEIGHT COMPLEXES BETWEEN BROWN ALGAL PHENOLIC SUBSTANCES AND ALGINATES1

The interaction between phenolic substances (PS) and alginates (ALG) has been suggested to play a role in the structure of the cell walls of brown seaweeds. However, no clear evidence for this interaction was reported. Vanadium bromoperoxidase (VBPO) has been proposed as a possible catalyst for the binding of PS to ALG. In this work, we studied the interaction between PS and ALG from brown algae using size exclusion chromatography (SEC) and optical tweezers microscopy. The analysis by SEC revealed that ALG forms a high‐molecular‐weight complex with PS. To study the formation of this molecular complex, we investigated the in vitro interaction of purified ALG from Fucus vesiculosus L. with purified PS from Padina gymnospora (Kütz.) Sond., in the presence or absence of VBPO. The interaction between PS and ALG only occurred when VBPO was added, indicating that the enzyme is essential for the binding process. The interaction of these molecules led to a reduction in ALG viscosity. We propose that VBPO promotes the binding of PS molecules to the ALG uronic acids residues, and we also suggest that PS are components of the brown algal cell walls.     

Relationship Among Phenolic Contents,Seed Predation,and Physical Seed Traits in <Emphasis Type="Italic">Mimosa bimucronata</Emphasis> Plants

Phenolic contents were compared between Mimosa bimucronata seeds from infested and non-infested fruits to assess induced defense response. By measuring leg length of the bruchid beetle Acanthoscelides schrankiae, we verified whether phenolic contents affected bruchid body size. In addition, the relationship between physical seed traits and phenolic contents was examined. Results showed that seeds from infested fruits had significantly greater phenolic contents than seeds from non-infested fruits, which suggested induced defense. Body size variation in A. schrankiae was marginally nonsignificant according to phenolic contents among plants (negative trend), indicating that phenols may interfere directly with bruchid performance. Seeds that were more irregularly shaped had significantly greater phenolic contents than those that were more uniform. Therefore, the most perfectly spherical seeds may be more vulnerable to seed predation, and our results suggest that the production of phenolic compounds was increased in infested fruits, which in turn may affect A. schrankiae development.     

Arrest of oogenesis in the bug Rhodnius prolixus challenged with the fungus Aspergillus niger is mediated by immune response-derived PGE2

In this work we characterized the immune response of the insect Rhodnius prolixus to a direct injection into the hemocoel of the non-entomopathogenic fungus Aspergillus niger, and evaluated its consequences on host oogenesis. These animals were able to respond by mounting effective cellular and humoral responses to this fungus; these responses were shown, however, to have reproductive fitness costs, as the number of eggs laid per female was significantly reduced. The disturbance of egg formation during infectious process correlated with an elevation in the titer of hemolymph prostaglandin E₂ 48 h post-challenge. Administration of Zymosan A as an immunogenic non-infectious challenge produced similar effects on phenoloxidase and prophenoloxidase activities, oocyte development and prostaglandin E₂ titer, precluding the hypothesis of an effect mediated by fungal metabolites in animals challenged with fungus. Ovaries at 48 h post-challenge showed absence of vitellogenic ovarian follicles, and the in vivo administration of prostaglandin E₂ or its receptor agonist misoprostol, partially reproduced this phenotype. Together these data led us to hypothesize that immune-derived prostaglandin E₂ raised from the insect response to the fungal challenge is involved in disturbing follicle development, contributing to a reduction in host reproductive output and acting as a host-derived adaptive effector to infection.     

Immunohistochemical detection of polyductin and co-localization with liver progenitor cell markers during normal and abnormal development of the intrahepatic biliary system and in adult hepatobiliary carcinomas

The longest open reading frame of PKHD1 (polycystic kidney and hepatic disease 1), the autosomal recessive polycystic kidney disease (ARPKD) gene, encodes a single-pass, integral membrane protein named polyductin or fibrocystin. A fusion protein comprising its intracellular C-terminus, FP2, was previously used to raise a polyclonal antiserum shown to detect polyductin in several human tissues, including liver. In the current study, we aimed to investigate by immunohistochemistry the detailed polyductin localization pattern in normal (ductal plate [DP], remodelling ductal plate [RDP], remodelled bile ducts) and abnormal development of the primitive intrahepatic biliary system, known as ductal plate malformation (DPM). This work also included the characterization of polyductin expression profile in various histological forms of neonatal and infantile cholestasis, and in cholangiocellular carcinoma (CCC) and hepatocellular carcinoma (HCC). We detected polyductin expression in the intrahepatic biliary system during the DP and the RDP stages as well as in DPM. No specific staining was found at the stage of remodelled bile ducts. Polyductin was also detected in liver biopsies with neonatal cholestasis, including mainly biliary atresia and neonatal hepatitis with ductular reaction as well as congenital hepatic fibrosis. In addition, polyductin was present in CCC, whereas it was absent in HCC. Polyductin was also co-localized in some DP cells together with oval stem cell markers. These results represent the first systematic study of polyductin expression in human pathologies associated with abnormal development of intrahepatic biliary tree, and support the following conclusions: (i) polyductin expression mirrors developmental properties of the primitive intrahepatic biliary system; (ii) polyductin is re-expressed in pathological conditions associated with DPM and (iii) polyductin might be a potential marker to distinguish CCC from HCC.