AsKC11, a Kunitz Peptide from Anemonia sulcata,Is a Novel Activator of G Protein-Coupled Inward-Rectifier Potassium Channels

(1) Background: G protein-coupled inward-rectifier potassium (GIRK) channels, especially neuronal GIRK1/2 channels, have been the focus of intense research interest for developing drugs against brain diseases. In this context, venom peptides that selectively activate GIRK channels can be seen as a new source for drug development. Here, we report on the identification and electrophysiological characterization of a novel activator of GIRK1/2 channels, AsKC11, found in the venom of the sea anemone Anemonia sulcata. (2) Methods: AsKC11 was purified from the sea anemone venom by reverse-phase chromatography and the sequence was identified by mass spectrometry. Using the two-electrode voltage-clamp technique, the activity of AsKC11 on GIRK1/2 channels was studied and its selectivity for other potassium channels was investigated. (3) Results: AsKC11, a Kunitz peptide found in the venom of A. sulcata, is the first peptide shown to directly activate neuronal GIRK1/2 channels independent from Gi/o protein activity, without affecting the inward-rectifier potassium channel (IRK1) and with only a minor effect on K_V1.6 channels. Thus, AsKC11 is a novel activator of GIRK channels resulting in larger K⁺ currents because of an increased chord conductance. (4) Conclusions: These discoveries provide new insights into a novel class of GIRK activators.     

A Tale of Toxin Promiscuity: The Versatile Pharmacological Effects of Hcr 1b-2 Sea Anemone Peptide on Voltage-Gated Ion Channels

Sea anemones are a rich source of biologically active compounds. Among approximately 1100 species described so far, Heteractis crispa species, also known as sebae anemone, is native to the Indo-Pacific area. As part of its venom components, the Hcr 1b-2 peptide was first described as an ASIC1a and ASIC3 inhibitor. Using Xenopus laevis oocytes and the two-electrode voltage-clamp technique, in the present work we describe the remarkable lack of selectivity of this toxin. Besides the acid-sensing ion channels previously described, we identified 26 new targets of this peptide, comprising 14 voltage-gated potassium channels, 9 voltage-gated sodium channels, and 3 voltage-gated calcium channels. Among them, Hcr 1b-2 is the first sea anemone peptide described to interact with isoforms from the Kv7 family and T-type Cav channels. Taken together, the diversity of Hcr 1b-2 targets turns this toxin into an interesting tool to study different types of ion channels, as well as a prototype to develop new and more specific ion channel ligands.     

Pronuclear Embryo Yield in Canindé and Saanen Goats for DNA Microinjection

The objective of this study was to examine the effect of donor breed on pronuclear‐stage embryo yield to be used for DNA microinjection in a transgenesis goat program. Twelve Canindé and twelve Saanen goats were heat synchronized using a progestagen‐cloprostenol treatment. Forty‐eight hours before the sponge removal, superovulation was induced with a total administration of 4.4 mg/kg bodyweight NIH‐FSH‐P1, given twice daily in decreasing doses over 3 days. In addition, goats received 100 μg of GnRH and they were hand‐mated at 36 and 48 h after progestagen removal. Embryo recovery was performed by oviduct flushing at 72 h after sponge removal. Embryos were microinjected with a DNA construct and noticeable swelling of the nuclei was the criterion for successful microinjection. The total diameter, cytoplasm diameter, zona pellucida thickness and pronuclei diameter were measured for each microinjected embryo. A higher (p < 0.05) percentage of fertilized ova was observed in Canindé (89.9%) than Saanen (36.2%) goats. In addition, Canindé donors produced a higher percentage of pronuclear embryos when compared with Saanen: 72.5% vs 20.6% (p < 0.05), respectively. Successful microinjection was verified in 96.7% and 73.3% of times in Canindé and Saanen embryos, respectively (p < 0.05). Significant differences were observed for all morphometric parameters except for cytoplasm diameter. In conclusion, under our study experimental conditions, Canindé were more efficient than Saanen goats concerning the pronuclear embryo yield and manipulation. The use of Canindé goats in transgenesis programs could be increase the interest in their breeding and could be contribute to saving them from extinction.     

Secreted phosphatase activities in trypanosomatid parasites of plants modulated by platelet-activating factor

ABSTRACT The secreted phosphatase activities of two trypanosomatid parasites were characterized and compared with supernatants of living cells. The plant parasite Phytomonas fran?ai and the phytophagous hemipteran parasite Herpetomonas sp. hydrolyzed p-nitrophenylphosphate at a rate of 15.54 and 6.51 nmol Pi/mg of protein per min, respectively. Sodium orthovanadate (N(a)VO(3)) and sodium fluoride (NaF) decreased the phosphatase activities. The phosphatase activity of P. fran?ai was drastically diminished (73% inhibition) in the presence of sodium tartrate, whereas the phosphatase activity of Herpetomonas sp. was inhibited by 23%. Cytochemical analysis showed the localization of these enzymes on the external surface and in the flagellar pocket of the two trypanosomatids. Sodium tartrate inhibited this reaction, confirming the biochemical data. Platelet-activating factor modulated the phosphatase activities, inhibiting P. fran?ai activity and stimulating Herpetomonas sp. phosphatase activity.     

Survival of Ornithobacterium rhinotracheale in sterilized poultry litter

Ornithobacterium rhinotracheale (ORT) has been associated with respiratory disease, increased mortality, retarded growth, and decreased egg production in chickens and turkeys. Surveillance of exposure to ORT infection in the field has shown that prevalence of the infection is higher during winter months. The ability of ORT to remain viable in the poultry litter was studied at different temperatures over time. Presterilized poultry litter was inoculated with 10(11) colony-forming units of ORT and kept at -12 C, 4 C, 22 C, 37 C, and 42 C. Reisolation and titration of ORT from litter was attempted at intervals. Results indicate that ORT survived for 1 day at 37 C, 6 days at 22 C, 40 days at 4 C, and at least 150 days at -12 C. ORT did not survive 24 hr at 42 C. The survival of ORT at lower temperatures may be associated with the higher incidence of ORT infection in poultry during winter months.     

Outer membrane proteins for serologic detection of Ornithobacterium rhinotracheale infection in turkeys

Ornithobacterium rhinotracheale (ORT) is a bacterium responsible for a respiratory disease in turkeys and chickens and has been identified as one of the emerging respiratory bacterial pathogens. The clinical signs and lesions caused by ORT are very similar to those caused by other respiratory infectious agents; therefore, an accurate diagnostic test is necessary to identify the infection. In this study, we have investigated the use of outer membrane proteins of ORT in an indirect enzyme-linked immunosorbent assay (ELISA) to detect the exposure to ORT infection. Outer membrane proteins of ORT were extracted and used as an antigen in ELISA to detect infection in turkeys exposed to different serotypes of ORT. The ELISA results were compared with the conventional serum plate agglutination test. The agglutination test detected specific antibodies for ORT in 65% of experimentally infected turkeys during the first 2 wk of infection. The ELISA detected up to 100% of infected birds for 8 wk postinfection. The results suggest that ELISA is able to detect the exposure to ORT in later stages of the infection and this assay can be used in serologic surveillance of ORT infection for poultry in the field.     

Effects of deficit irrigation strategies on cluster microclimate for improving fruit composition of Moscatel field-grown grapevines

The grapevine plays a very important role in the economic, social and cultural sectors of many regions; however vineyards are often grown in regions under stressful conditions and thus they are vulnerable to climate change. The objective of this research was to investigate the effect of partial root-zone drying (PRD) irrigation on vine water relations, vegetative growth, plant microclimate, berry composition and yield components, compared to conventional deficit irrigation (DI, 50% ETc), full irrigation (FI, 100% of ETc) and non-irrigated vines (NI). The study was undertaken in mature ‘Moscatel’ grapevines (Vitis vinifera L.) grown in Pegões, South of Portugal. Compared to the other irrigated treatments, PRD vines showed a better microclimate at the cluster zone with higher incident photosynthetic photon flux density (PPFD). Within the more open canopies of NI and PRD treatments, berry temperatures were higher than those of denser ones (DI and FI). Compared to the conventional irrigation technique the better microclimate observed in PRD vines was a consequence of a reduction in vine growth, where lower values of leaf layer number, leaf area, canopy wideness, water shoots and shoot weight were observed. In PRD vines we observed a tendency to a development of a deeper root system, while DI and FI showed a more homogeneous root distribution throughout the different soil layers. PRD showed an improvement in berry quality with higher values of flavour precursors, and total phenols concentration without any significant yield reduction compared to DI and FI.     

Ineffectiveness of pruning to control citrus huanglongbing caused by <Emphasis Type="Italic">Candidatus</Emphasis> Liberibacter americanus

The huanglongbing (HLB) disease of citrus trees, caused by Candidatus Liberibacter asiaticus and Ca. Liberibacter americanus, was first reported in Brazil in March, 2004. The presence of the disease has caused serious concerns among growers. Pruning experiments were conducted to determine if removal of symptomatic branches or the entire canopy (decapitation) would eliminate infected tissues and save HLB-affected trees. Pruning was done in five blocks on a total of 592 3- to 16 year-old ‘Valência’, ‘Hamlin’ or ‘Pêra’ sweet orange trees showing no symptoms or with two levels of symptom severity. Ten decapitated trees per block were caged and all trees were treated with insecticides to control the psyllid vector, Diaphorina citri. Mottled leaves reappeared on most symptomatic (69.2%) as well on some asymptomatic (7.6%) pruned trees, regardless of age, variety, and pruning procedure. Presence of the pathogen (Ca. Liberibacter americanus) in all symptomatic trees was confirmed by PCR. In general, the greater the symptom severity before pruning the lower the percentage of trees that remained asymptomatic after pruning.     

温度对大叶桃花心木(Swietenia macrophylla King)幼苗叶片的光合影响

本文研究了大叶桃花心木（Swietenia macrophylla King）一年生幼苗在经过夜温处理后的光响应曲线和在饱和光强下的CO2反应曲线.结果表明:在大气CO2浓度下,叶片的最佳光合作用温度在25-31℃之间,而在饱和CO2浓度下为31-35℃.在25℃以下光合速率开始降低,主要是由于羧化效率的降低,而当温度超过31℃时,光合速率下降,是因为羧化效率的降低和呼吸速率的增加.CO2浓度对光合的促进作用在低温下受到抑制,这意味着未来在CO2浓度增高的情况下,高浓度的CO2对热带常绿植物光合的促进在冬天低温情况下表现不十分明显.图4参23.     

Milk production,intake, digestion,blood parameters,and ingestive behavior of cows supplemented with by-products from the biodiesel industry

This study aimed to evaluate the intake, digestion, blood parameters, and feeding behavior of crossbred dairy cows (Holstein?×?Gir) managed on Panicum maximum Jacq. cv. Tanzania-1 and provided supplementation with groundnut cake, sunflower cake, or palm kernel cake (to replace soybean meal). Sixteen cows were randomly assigned in a Latin square design with four treatments and four experimental periods. The consumption of nutrients from the pasture did not vary between experimental treatments. Cows receiving the palm kernel cake supplement had a reduced crude protein, non-fibrous carbohydrate, and total digestible nutrient intake and an increase in the average consumption of ether extract. There was also a reduction in the digestibility of dry matter. The inclusion of supplements in the diet did not influence the average time spent eating, ruminating, or resting. The mean values of respiratory and heart rates showed thermal comfort during the trial period. There was a reduction in blood urea nitrogen with palm kernel cake supplementation, and the values of total protein, albumin, and glucose were also significantly different with this supplement. It is recommended that cakes of groundnut cake and sunflower cake seed be used for a total replacement of soybean meal supplements for lactating cows, but the use of palm kernel cake is not recommended.