Leucokinin and Associated Neuropeptides Regulate Multiple Aspects of Physiology and Behavior in Drosophila

Leucokinins (LKs) constitute a family of neuropeptides identified in numerous insects and many other invertebrates. LKs act on G-protein-coupled receptors that display only distant relations to other known receptors. In adult Drosophila, 26 neurons/neurosecretory cells of three main types express LK. The four brain interneurons are of two types, and these are implicated in several important functions in the fly’s behavior and physiology, including feeding, sleep–metabolism interactions, state-dependent memory formation, as well as modulation of gustatory sensitivity and nociception. The 22 neurosecretory cells (abdominal LK neurons, ABLKs) of the abdominal neuromeres co-express LK and a diuretic hormone (DH44), and together, these regulate water and ion homeostasis and associated stress as well as food intake. In Drosophila larvae, LK neurons modulate locomotion, escape responses and aspects of ecdysis behavior. A set of lateral neurosecretory cells, ALKs (anterior LK neurons), in the brain express LK in larvae, but inconsistently so in adults. These ALKs co-express three other neuropeptides and regulate water and ion homeostasis, feeding, and drinking, but the specific role of LK is not yet known. This review summarizes Drosophila data on embryonic lineages of LK neurons, functional roles of individual LK neuron types, interactions with other peptidergic systems, and orchestrating functions of LK.     

Structural Aspects and Prediction of Calmodulin-Binding Proteins

Calmodulin (CaM) is an important intracellular protein that binds Ca²⁺ and functions as a critical second messenger involved in numerous biological activities through extensive interactions with proteins and peptides. CaM’s ability to adapt to binding targets with different structures is related to the flexible central helix separating the N- and C-terminal lobes, which allows for conformational changes between extended and collapsed forms of the protein. CaM-binding targets are most often identified using prediction algorithms that utilize sequence and structural data to predict regions of peptides and proteins that can interact with CaM. In this review, we provide an overview of different CaM-binding proteins, the motifs through which they interact with CaM, and shared properties that make them good binding partners for CaM. Additionally, we discuss the historical and current methods for predicting CaM binding, and the similarities and differences between these methods and their relative success at prediction. As new CaM-binding proteins are identified and classified, we will gain a broader understanding of the biological processes regulated through changes in Ca²⁺ concentration through interactions with CaM.     

Contribution of Invariant Natural Killer T Cells to the Clearance of Pseudomonas aeruginosa from Skin Wounds

Chronic infections are considered one of the most severe problems in skin wounds, and bacteria are present in over 90% of chronic wounds. Pseudomonas aeruginosa is frequently isolated from chronic wounds and is thought to be a cause of delayed wound healing. Invariant natural killer T (iNKT) cells, unique lymphocytes with a potent regulatory ability in various inflammatory responses, accelerate the wound healing process. In the present study, we investigated the contribution of iNKT cells in the host defense against P. aeruginosa inoculation at the wound sites. We analyzed the re-epithelialization, bacterial load, accumulation of leukocytes, and production of cytokines and antimicrobial peptides. In iNKT cell–deficient (Jα18KO) mice, re-epithelialization was significantly decreased, and the number of live colonies was significantly increased, when compared with those in wild-type (WT) mice on day 7. IL-17A, and IL-22 production was significantly lower in Jα18KO mice than in WT mice on day 5. Furthermore, the administration of α-galactosylceramide (α-GalCer), a specific activator of iNKT cells, led to enhanced host protection, as shown by reduced bacterial load, and to increased production of IL-22, IL-23, and S100A9 compared that of with WT mice. These results suggest that iNKT cells promote P. aeruginosa clearance during skin wound healing.     

In Vivo Electrophysiology of Peptidergic Neurons in Deep Layers of the Lumbar Spinal Cord after Optogenetic Stimulation of Hypothalamic Paraventricular Oxytocin Neurons in Rats

The spinal ejaculation generator (SEG) is located in the central gray (lamina X) of the rat lumbar spinal cord and plays a pivotal role in the ejaculatory reflex. We recently reported that SEG neurons express the oxytocin receptor and are activated by oxytocin projections from the paraventricular nucleus of hypothalamus (PVH). However, it is unknown whether the SEG responds to oxytocin in vivo. In this study, we analyzed the characteristics of the brain–spinal cord neural circuit that controls male sexual function using a newly developed in vivo electrophysiological technique. Optogenetic stimulation of the PVH of rats expressing channel rhodopsin under the oxytocin receptor promoter increased the spontaneous firing of most lamina X SEG neurons. This is the first demonstration of the in vivo electrical response from the deeper (lamina X) neurons in the spinal cord. Furthermore, we succeeded in the in vivo whole-cell recordings of lamina X neurons. In vivo whole-cell recordings may reveal the features of lamina X SEG neurons, including differences in neurotransmitters and response to stimulation. Taken together, these results suggest that in vivo electrophysiological stimulation can elucidate the neurophysiological response of a variety of spinal neurons during male sexual behavior.     

Characteristic tryptic peptides and gelling properties of porcine skin gelatin affected by thermal action

Thermal action in extraction process had effects on characteristic tryptic peptides identification and gelling properties of porcine gelatin. SDS-PAGE, HPLC-LTQ/Orbitrap high-resolution mass spectrometry, texture analyser and rheometer were used to evaluate collagen depolymerisation degree, characteristic tryptic peptides and gelling properties of gelatins prepared in various thermal actions. Results showed that with increasing temperature and time, depolymerisation degree enlarged, while gel strength, gelling and melting temperature decreased. Mass spectra showed that 47 and 49 common characteristic tryptic peptides were identified in gelatins extracted at 50 °C and 100 °C with various times, respectively. Moreover, 34 common characteristic tryptic peptides were identified in all gelatin samples. Further comparison between this work and our previous investigations yielded 20 common characteristic tryptic peptides, which stably exist in various thermal actions. These common characteristic tryptic peptides may be very helpful for the accurate authentication of porcine gelatin.     

微波辅助孵育法富集麦胚多肽的工艺优化

目的 麦胚孵育过程中蛋白酶将蛋白质水解成肽和氨基酸，多肽具有明确的生理活性和营养调节作用。本研究以麦胚为试验原料，采用微波辅助预处理的方法，研究孵育的温度、时间、pH及料液比4种因素对孵育后的蛋白酶活力及多肽含量的影响。 方法 通过单因素和响应曲面试验进行工艺条件优化。 结果 与未进行微波辅助预处理的样品相比，微波辅助处理（600 W, 10 s）能显著提高孵育后样品中的蛋白酶酶活力（约9.4倍）和肽含量（约3.1倍）。经过微波辅助处理后，孵育温度为51.5℃、pH为4.0、时间为6.33 h、料液比为1:7时，蛋白酶活力达最高为3826.24 U/g；孵育温度为45.0 ℃、pH为4.8、时间为8 h、料液比为1:7时，肽含量达最高为262.63 mg/g。 结论 微波辅助处理能有效的激活麦胚孵育液中的内源性蛋白酶活性，促进蛋白水解反应，显著提高孵育后的肽含量。该研究结果为麦胚多肽的制备新工艺开发提供了研究基础。     

Cheese ripening in nonconventional conditions: A multiparameter study applied to Protected Geographical Indication Canestrato di Moliterno cheese

A multiparameter study was performed to evaluate the effect of fondaco, a traditional ripening cellar without any artificial temperature and relative humidity control, on the chemical, microbiological, and sensory characteristics of Protected Geographical Indication Canestrato di Moliterno cheese. Ripening in such a nonconventional environment was associated with lower counts of lactococci, lactobacilli, and total viable bacteria, and higher presence of enterococci, in comparison with ripening in a controlled maturation room. Moreover, fondaco cheese underwent accelerated maturation, as demonstrated by faster casein degradation, greater accumulation of free AA, and higher formation of volatile organic compounds. Secondary proteolysis, as assessed by liquid chromatography-mass spectrometry of free AA and low molecular weight peptides, did not show any qualitative difference among cheeses, but fondaco samples evidenced an advanced level of peptidolysis. On the other hand, significant qualitative differences were observed in the free fatty acid profiles and in the sensory characteristics. Principal component analysis showed a clear separation of the fondaco and control cheeses, indicating that ripening in the natural room conferred unique sensory features to the product.     

石油降解菌ZS1对典型抗生素毒害作用的应激响应机制

为了解抗生素对石油降解菌的生态效应及微生物对典型抗生素毒害作用的应激响应机制，通过开展不同浓度典型抗生素对高效石油降解菌铜绿假单胞菌（ZS1）的毒性试验，研究了菌体细胞在3种不同种类抗生素（土霉素、红霉素、磺胺嘧啶）作用下的生长及毒理指标。结果表明，3种抗生素对ZS1有不同的抑菌效果，其中土霉素对ZS1活性的抑制程度最强，抑制率随着土霉素浓度的增大而明显上升，有显著的浓度-效应关系。研究了ZS1抗氧化酶（过氧化氢酶、过氧化物酶、超氧化物歧化酶）对抗生素的响应机制，结果表明ZS1受到污染时会通过产生抗氧化酶抵抗外界污染并维持细胞功能。     

Germination improves the functional properties of soybean and enhances soymilk quality

The effects of soybean variety and germination time on structural changes, antinutritional factor content, antioxidant activity of germinated soybean, and on the functional properties of soymilk were comprehensively investigated. The results showed that the antioxidant activity increased with increasing germination time. The content of antinutritional factors decreased with increasing germination time. Soybean varieties with the lowest tannin and trypsin inhibitor content were DN690 and HJ1. Lipoxygenase activity and phytic acid content showed no significant differences among soybean varieties. The content of α-helices and β-turns in soybean protein decreased with increasing germination time, while the content of β-sheets was increased. Soybean protein was progressively broken down into smaller molecular peptides during the germination process. The digestion and content of soluble protein in soymilk increased with germination. In summary, we show that germination is an effective, cheap, and green method that improves the functional properties of soybean and soymilk.