Most dynorphin neurons in the zona incerta-perifornical area are active in waking relative to non-rapid-eye movement and rapid-eye movement sleep.

Dynorphin is an endogenous opiate localized in many brain regions and spinal cord, but the activity of dynorphin neurons during sleep is unknown. Dynorphin is an inhibitory neuropeptide that is coreleased with orexin, an excitatory neuropeptide. We used microendoscopy to test the hypothesis that, like orexin, the dynorphin neurons are wake-active. Dynorphin-cre mice (n = 3) were administered rAAV8-Ef1a-Con/Foff 2.0-GCaMP6M into the zona incerta-perifornical area, implanted with a GRIN lens (gradient reflective index), and electrodes to the skull that recorded sleep. One month later, a miniscope imaged calcium fluorescence in dynorphin neurons during multiple bouts of wake, non-rapid-eye movement (NREM), and rapid-eye movement (REM) sleep. Unbiased data analysis identified changes in calcium fluorescence in 64 dynorphin neurons. Most of the dynorphin neurons (72%) had the highest fluorescence during bouts of active and quiet waking compared to NREM or REM sleep; a subset (20%) were REM-max. Our results are consistent with the emerging evidence that the activity of orexin neurons can be classified as wake-max or REM-max. Since the two neuropeptides are coexpressed and coreleased, we suggest that dynorphin-cre-driven calcium sensors could increase understanding of the role of this endogenous opiate in pain and sleep.

Unilateral optogenetic stimulation of Lhx6 neurons in the zona incerta increases REM sleep.

To determine how a waking brain falls asleep researchers have monitored and manipulated activity of neurons and glia in various brain regions. While imaging Gamma-Aminobutyric Acid (GABA) neurons in the zona incerta (ZI) we found a subgroup that anticipates onset of NREM sleep (Blanco-Centurion C, Luo S, Vidal-Ortiz A, Swank C, Shiromani PJ. Activity of a subset of vesicular GABA-transporter neurons in the ventral ZI anticipates sleep onset. Sleep. 2021;44(6):zsaa268. doi:10.1093/sleep/zsaa268.). To differentiate the GABA subtype we now image and optogenetically manipulate the ZI neurons containing the transcription factor, Lhx6. In the first study, Lhx6-cre mice (n = 5; female = 4) were given rAAV-DJ-EF1a-DIO-GCaMP6M into the ZI (isofluorane anesthesia), a GRIN lens implanted, and 21days later sleep and fluorescence in individual Lhx6 neurons were recorded for 4 hours. Calcium fluorescence was detected in 132 neurons. 45.5% of the Lhx6 neurons were REM-max; 30.3% were wake-max; 11.4% were wake + REM max; 9% were NREM-max; and 3.8% had no change. The NREM-max group of neurons fluoresced 30 seconds ahead of sleep onset. The second study tested the effects of unilateral optogenetic stimulation of the ZI Lhx6 neurons (n = 14 mice) (AAV5-Syn-FLEX-rc[ChrimsonR-tdTomato]. Stimulation at 1 and 5 Hz (1 minute on- 4 minutes off) significantly increased percent REM sleep during the 4 hours stimulation period (last half of day cycle). The typical experimental approach is to stimulate neurons in both hemispheres, but here we found that low-frequency stimulation of ZI Lhx6 neurons in one hemisphere is sufficient to shift states of consciousness. Detailed mapping combined with mechanistic testing is necessary to identify local nodes that can shift the brain between wake-sleep states.

Phenylpropanoid Derivatives and Their Role in Plants' Health and as antimicrobials.

Phenylpropanoids belong to a wide group of compounds commonly secreted by plants and involved in different roles related with plant growth and development and the defense against plant pathogens. Some key intermediates from shikimate pathway are used to synthesize these compounds. In this way, by the phenylpropanoid pathway several building blocks are achieved to obtain flavonoids, isoflavonoids, coumarins, monolignols, phenylpropenes, phenolic acids, stilbenes and stilbenoids, and lignin, suberin and sporopollenin for plant-microbe interactions, structural support and mechanical strength, organ pigmentation, UV protection and acting against pathogens. Some reviews have revised phenylpropanoid biosynthesis and regulation of the biosynthetic pathways. In this review, the most important chemical structures about phenylpropanoid derivatives are summarized grouping them in different sections according to their structure. We have put special attention on their different roles in plants especially in plant health, growth and development and plant-environment interactions. Their interaction with microorganisms is discussed including their role as antimicrobials. We summarize all new findings about new developed structures and their involvement in plants health.

The possibility of using Serratia isolates for the production of biopreparations in the protection of plants against diseases and pests.

The agriculture is extremely important for the life of human beings. Therefore, it is needed to control the enemies that destroy vast areas of crops causing great economic losses. Chemical pesticides were the option for many decades, but the damage that they cause to environment and human health led to the idea of changing the use of these for more sustainable options such as biopesticides as a biological control. Among microbial pesticides, Serratia species have been found as suitable options to apply against several pests or pathogens. Serratia species produce a wide range of secondary metabolites with several biological activities such as antifungal, antibacterial, and pesticides which can be used in sustainable agriculture. It has been reported that several Serratia species are able to suppress some crop diseases caused by Fusarium oxysporum, Rhizoctonia solani, Phytophthora parasitica, Sclerotinia sclerotiorum, Verticillium dahlia, and Phytophthora capsici among others. Therefore, they have been used as biocontrol agents in agriculture. In this review, we summarized the genus Serratia describing its history and development and the metabolites it secretes, which are responsible for their antibacterial and antifungal activity. We have analyzed the insecticide capacity of several Serratia species as well antifungal properties of Serratia species against most important crops' pathogens. In conclusion, the use of Serratia as a biological control agent against plant pathogens can be a good option for a sustainable agriculture. More work is needed to assess the safety of the isolated new strains and their effectiveness against pathogens in in vivo conditions.

Bacillus spp. as Bio-factories for Antifungal Secondary Metabolites: Innovation Beyond Whole Organism Formulations.

Several fungi act as parasites for crops causing huge annual crop losses at both pre- and post-harvest stages. For years, chemical fungicides were the solution; however, their wide use has caused environmental contamination and human health problems. For this reason, the use of biofungicides has been in practice as a green solution against fungal phytopathogens. In the context of a more sustainable agriculture, microbial biofungicides have the largest share among the commercial biocontrol products that are available in the market. Precisely, the genus Bacillus has been largely studied for the management of plant pathogenic fungi because they offer a chemically diverse arsenal of antifungal secondary metabolites, which have spawned a heightened industrial engrossment of it as a biopesticide. In this sense, it is indispensable to know the wide arsenal that Bacillus genus has to apply these products for sustainable agriculture. Having this idea in our minds, in this review, secondary metabolites from Bacillus having antifungal activity are chemically and structurally described giving details of their action against several phytopathogens. Knowing the current status of Bacillus secreted antifungals is the base for the goal to apply these in agriculture and it is addressed in depth in the second part of this review.

Activity of GABA neurons in the zona incerta and ventral lateral periaqueductal grey is biased towards sleep.

STUDY OBJECTIVES: As in various brain regions the activity of gamma-aminobutyric acid (GABA) neurons is largely unknown, we measured in vivo changes in calcium fluorescence in GABA neurons in the zona incerta (ZI) and the ventral lateral periaqueductal grey (vlPAG), two areas that have been implicated in regulating sleep. METHODS: vGAT-Cre mice were implanted with sleep electrodes, microinjected with rAAV-DIO-GCaMP6 into the ZI (n = 6) or vlPAG (n = 5) (isoflurane anesthesia) and a GRIN (Gradient-Index) lens inserted atop the injection site. Twenty-one days later, fluorescence in individual vGAT neurons was recorded over multiple REM cycles. Regions of interest corresponding to individual vGAT somata were automatically extracted with PCA-ICA analysis. RESULTS: In the ZI, 372 neurons were identified. Previously, we had recorded the activity of 310 vGAT neurons in the ZI and we combined the published dataset with the new dataset to create a comprehensive dataset of ZI vGAT neurons (total neurons = 682; mice = 11). In the vlPAG, 169 neurons (mice = 5) were identified. In both regions, most neurons were maximally active in REM sleep (R-Max; ZI = 51.0%, vlPAG = 60.9%). The second most abundant group was W-Max (ZI = 23.9%, vlPAG = 25.4%). In the ZI, but not in vlPAG, there were neurons that were NREMS-Max (11.7%). vlPAG had REMS-Off neurons (8.3%). In both areas, there were two minor classes: wake/REMS-Max and state indifferent. In the ZI, the NREMS-Max neurons fluoresced 30 s ahead of sleep onset. CONCLUSIONS: These descriptive data show that the activity of GABA neurons is biased in favor of sleep in two brain regions implicated in sleep.

A study of bacteriocin like substances comparison produced by different species of Bacillus related to B. cereus group with specific antibacterial activity against foodborne pathogens.

The antibiotic-resistant bacteria are emerging as a great threat worldwide. For this reason it is important to develop new antibiotic substances. Bacillus is considered as a factory of a wide range of chemical compounds with a variety of activities. Among these substances are bacteriocins which are small peptides showing stability in a wide range of pH and temperatures and having a potent antibacterial activity. Bacillus species can be grouped into families such as B. cereus group based on their genetic similarity. It can be helpful to study the bacteriocins presented in these related species identifying the differences and similarities between them to relate the presence of a given bacteriocin with the producer specie. The aim of this study was to isolate the bacteriocins from three related species of B. cereus group such as B. mycoides, B. weihenstephanensis and B. toyonensis and compare among them and with the bacteriocins isolated from B. velezensis. Besides it was analyzed the bactericidal activity of each isolated bacteriocin. Five different bacteriocins of similar molecular mass and specific against foodborne pathogens were isolated from three Bacillus species related to B. cereus group, that were quite different both in molecular mass and bactericidal activity from that was isolated from B. velezensis. The results indicated that bacteriocins can be distinguished according to Bacillus specie from it has been isolated.

Asymmetric Organocatalytic Syntheses of Bioactive Compounds.

BACKGROUND: The total syntheses of complex natural products have evolved to include new methodologies to save time, simplifying the form to achieve these natural compounds. OBJECTIVES: In this review, we have described the asymmetric synthesis of different natural products and biologically active compounds of the last ten years until the current day. RESULTS: An asymmetric organocatalytic reaction is a key to generate stereoselectively the main structure with the required stereochemistry. CONCLUSION: Even more remarkable, the organocatalytic cascade reactions, which are carried out with high stereoselectivity, as well as a possible approximation of the organocatalysts activation with substrates are also described.

Biosynthesis and beneficial effects of microbial gibberellins on crops for sustainable agriculture.

Soil microbes promote plant growth through several mechanisms such as secretion of chemical compounds including plant growth hormones. Among the phytohormones, auxins, ethylene, cytokinins, abscisic acid and gibberellins are the best understood compounds. Gibberellins were first isolated in 1935 from the fungus Gibberella fujikuroi and are synthesized by several soil microbes. The effect of gibberellins on plant growth and development has been studied, as has the biosynthesis pathways, enzymes, genes and their regulation. This review revisits the history of gibberellin research highlighting microbial gibberellins and their effects on plant health with an emphasis on the early discoveries and current advances that can find vital applications in agricultural practices.

Bacillus sp. Bacteriocins: Natural Weapons against Bacterial Enemies.

BACKGROUND: Currently antibiotic resistant pathogenic bacteria are emerging as an important health problem worldwide. The search for new compounds with antibiotic characteristics is the most promising alternative. Bacteriocins are natural compounds that are inhibitory against pathogens, and Bacillus species are the major producers of these compounds, showing antimicrobial activity against clinically important bacteria. These peptides not only have potential in the pharmaceutical industry but also in food and agricultural sectors. OBJECTIVE: We provide an overview of the recent bacteriocins isolated from different species of Bacillus including their applications and older bacteriocins. RESULTS: In this review, we have revised some works about the improvements carried out in the production of bacteriocins. CONCLUSION: These applications make bacteriocins very promising compounds that need to study for industrial production.

Mapping Network Activity in Sleep.

It was in the influenza pandemic of 1918 that von Economo identified specific brain regions regulating sleep and wake. Since then researchers have used a variety of tools to determine how the brain shifts between states of consciousness. In every enterprise new tools have validated existing data, corrected errors and made new discoveries to advance science. The brain is a challenge but new tools can disentangle the brain network. We summarize the newest tool, a miniature microscope, that provides unprecedented view of activity of glia and neurons in freely behaving mice. With this tool we have observed that the activity of a majority of GABA and MCH neurons in the lateral hypothalamus is heavily biased toward sleep. We suggest that miniscope data identifies activity at the cellular level in normal versus diseased brains, and also in response to specific hypnotics. Shifts in activity in small networks across the brain will help identify point of criticality that switches the brain from wake to sleep.

Recent advancements for microorganisms and their natural compounds useful in agriculture.

During the past years, microorganisms have been the cause of many problems for human's health. However, today with the development of many techniques of microbiology, the researchers have studied several roles of microorganisms which may help the society. Microbial-based products are expected to play important role in agriculture-enhancing plant production and therefore increasing crop's yieldeswani et al. . Microorganisms can act by several action mechanisms including antibiosis or mechanisms in plant-microbe interactions underlining the dual function of microbial strains toward plant nutrition and protection. The market has increased with the development of microbial-based products. Currently, it is normal to think that microorganisms help us in agriculture by applying them as biological control. In this mini review, we collect the last findings about this topic including very recent literature. KEY POINTS: • Microorganisms play a beneficial role in agriculture by different mechanisms. • One of these mechanisms is the secretion of chemical compounds with different activities.

Activity of a subset of vesicular GABA-transporter neurons in the ventral zona incerta anticipates sleep onset.

STUDY OBJECTIVES: Sleep and wake are opposing behavioral states controlled by the activity of specific neurons that need to be located and mapped. To better understand how a waking brain falls asleep it is necessary to identify activity of individual phenotype-specific neurons, especially neurons that anticipate sleep onset. In freely behaving mice, we used microendoscopy to monitor calcium (Ca2+) fluorescence in individual hypothalamic neurons expressing the vesicular GABA transporter (vGAT), a validated marker of GABA neurons. METHODS: vGAT-Cre mice (male = 3; female = 2) transfected with rAAV-FLEX-GCaMP6M in the lateral hypothalamus were imaged 30 days later during multiple episodes of waking (W), non-rapid-eye movement sleep (NREMS) or REMS (REMS). RESULTS: 372 vGAT neurons were recorded in the zona incerta. 23.9% of the vGAT neurons showed maximal fluorescence during wake (classified as wake-max), 4% were NREM-max, 56.2% REM-max, 5.9% wake/REM max, while 9.9% were state-indifferent. In the NREM-max group, Ca2+ fluorescence began to increase before onset of NREM sleep, remained high throughout NREM sleep, and declined in REM sleep. CONCLUSIONS: We found that 60.2% of the vGAT GABA neurons in the zona incerta had activity that was biased towards sleep (NREM and REMS). A subset of vGAT neurons (NREM-max) became active in advance of sleep onset and may induce sleep by inhibiting the activity of the arousal neurons. Abnormal activation of the NREM-max neurons may drive sleep attacks and hypersomnia.

A Strong Antifungal Activity of 7-O-Succinyl Macrolactin A vs Macrolactin A from Bacillus amyloliquefaciens ELI149.

Chemical analysis of the crude extract of bacterial strain Bacillus amyloliquefaciens ELI149, which had been previously isolated from soil, resulted in the isolation and characterization of two known macrolactin derivatives, macrolactin A (1) and 7-O-succinyl macrolactin A (2). The structures of two compounds were assigned by 1D/2D NMR techniques. The two compounds were demonstrated antifungal activity against some important phytopathogens. However, the presence of the succinyl moiety at C-7 gives to the molecule more activity being the second compound more active than the first, showing for the first time, a structure/activity relationship. The cellular damage was also studied in two important phytopathogen fungi.

Indole alkaloid derivatives as building blocks of natural products from Bacillus thuringiensis and Bacillus velezensis and their antibacterial and antifungal activity study.

Six known indole alkaloid derivatives have been isolated for the first time from Bacillus thuringiensis and Bacillus velezensis strains, all of them as building blocks for the synthesis of larger natural products. Their structure was elucidated by a complete spectroscopy. Their biological activities were tested against some Gram-positive and Gram-negative bacteria and three phytopathogenic fungi which cause diseases in important crops, such as Moniliophthora roreri, the causal agent of cacao disease. The results indicated that some compounds had modest antibacterial activity; however, some of them had strong antifungal activity against the probed fungi. This antifungal activity of these compounds has not been reported.

Macrolactin Antibiotics: Amazing Natural Products.

The resistance among various microbial species (infectious agents) to different antimicrobial drugs has emerged as a cause of serious public health problem all over the world. In this sense, natural products have been a rich source of compounds for drug discovery with antibiotic activity. Macrolactins are amazing structures which have antibiotic activity against some clinically relevant pathogens. In addition, they have anti-inflammatory, antifungal, antimicrobial, and antitumor activities. They are macrolides containing 24-membered lactone ring with some differences in their chemical structures. The synthesis of these compounds is a difficult task which has attracted attention of researchers; however few syntheses have been reported. In this review, the isolation of all reported macrolactins, their syntheses and biological activities are revisited.

The Chemistry of Drugs to Treat Candida albicans.

BACKGROUND: Candida species are in various parts of the human body as commensals. However, they can cause local mucosal infections and, sometimes, systemic infections in which Candida species can spread to all major organs and colonize them. OBJECTIVE: For the effective treatment of the mucosal infections and systemic life-threatening fungal diseases, a considerably large number of antifungal drugs have been developed and used for clinical purposes that comprise agents from four main drug classes: the polyenes, azoles, echinocandins, and antimetabolites. METHODS: The synthesis of some of these drugs is available, allowing synthetic modification of the molecules to improve the biological activity against Candida species. The synthetic methodology for each compound is reviewed. RESULTS: The use of these compounds has caused a high-level resistance against these drugs, and therefore, new antifungal substances have been described in the last years. The organic synthesis of the known and new compounds is reported. CONCLUSION: This article summarizes the chemistry of the existing agents, both the old drugs and new drugs, in the treatment of infections due to C. albicans, including the synthesis of the existing drugs.

Activity dynamics of amygdala GABAergic neurons during cataplexy of narcolepsy.

Recent studies showed activation of the GABAergic neurons in the central nucleus of the amygdala (CeA) triggered cataplexy of sleep disorder narcolepsy. However, there is still no direct evidence on CeA GABAergic neurons' real-time dynamic during cataplexy. We used a deep brain calcium imaging tool to image the intrinsic calcium transient as a marker of neuronal activity changes in the narcoleptic VGAT-Cre mice by expressing the calcium sensor GCaMP6 into genetically defined CeA GABAergic neurons. Two distinct GABAergic neuronal groups involved in cataplexy were identified: spontaneous cataplexy-ON and predator odor-induced cataplexy-ON neurons. Majority in the latter group were inactive during regular sleep/wake cycles but were specifically activated by predator odor and continued their intense activities into succeeding cataplexy bouts. Furthermore, we found that CeA GABAergic neurons became highly synchronized during predator odor-induced cataplexy. We suggest that the abnormal activation and synchronization of CeA GABAergic neurons may trigger emotion-induced cataplexy.

Dynamic Network Activation of Hypothalamic MCH Neurons in REM Sleep and Exploratory Behavior.

Most brain neurons are active in waking, but hypothalamic neurons that synthesize the neuropeptide melanin-concentrating hormone (MCH) are claimed to be active only during sleep, particularly rapid eye movement (REM) sleep. Here we use deep-brain imaging to identify changes in fluorescence of the genetically encoded calcium (Ca2+) indicator GCaMP6 in individual hypothalamic neurons that contain MCH. An in vitro electrophysiology study determined a strong relationship between depolarization and Ca2+ fluorescence in MCH neurons. In 10 freely behaving MCH-cre mice (male and female), the highest fluorescence occurred in all recorded neurons (n = 106) in REM sleep relative to quiet waking or non-REM sleep. Unexpectedly, 70% of the MCH neurons had strong fluorescence activity when the mice explored novel objects. Spatial and temporal mapping of the change in fluorescence between pairs of MCH neurons revealed dynamic activation of MCH neurons during REM sleep and activation of a subset of the same neurons during exploratory behavior. Functional network activity maps will facilitate comparisons of not only single-neuron activity, but also network responses in different conditions and disease.SIGNIFICANCE STATEMENT Functional activity maps identify brain circuits responding to specific behaviors, including rapid eye movement sleep (REM sleep), a sleep phase when the brain is as active as in waking. To provide the first activity map of individual neurons during REM sleep, we use deep-brain calcium imaging in unrestrained mice to map the activity of hypothalamic melanin-concentrating hormone (MCH) neurons. MCH neurons were found to be synchronously active during REM sleep, and also during the exploration of novel objects. Spatial mapping revealed dynamic network activation during REM sleep and activation of a subset of the neurons during exploratory behavior. Functional activity maps at the cellular level in specific behaviors, including sleep, are needed to establish a brain connectome.