Biology

Myotonic dystrophy type 1 (DM1) is a genetic disorder that causes muscle weakness and myotonia. In DM1 patients, cardiac electrical manifestations include conduction defects and atrial fibrillation. DM1 results in the expansion of a CTG transcribed into CUG-containing transcripts that accumulate in the nucleus as RNA foci and alter the activity of several splicing regulators. The underlying pathological mechanism involves two key RNA-binding proteins (MBNL and CELF) with expanded CUG repeats that sequester MBNL and alter the activity of CELF resulting in spliceopathy and abnormal electrical activity. In the present study, we identified two DM1 patients with heart conduction abnormalities and characterized their hiPSC lines. Two differentiation protocols were used to investigate both the ventricular and the atrial electrophysiological aspects of DM1 and unveil the impact of the mutation on voltage-gated ion channels, electrical activity, and calcium homeostasis in DM1 cardiomyocytes derived from hiPSCs. Our analysis revealed the presence of molecular hallmarks of DM1, including the accumulation of RNA foci and sequestration of MBNL1 in DM1 hiPSC-CMs. We also observed mis-splicing of SCN5A and haploinsufficiency of DMPK. Furthermore, we conducted separate characterizations of atrial and ventricular electrical activity, conduction properties, and calcium homeostasis. Both DM1 cell lines exhibited reduced density of sodium and calcium currents, prolonged action potential duration, slower conduction velocity, and impaired calcium transient propagation in both ventricular and atrial cardiomyocytes. Notably, arrhythmogenic events were recorded, including both ventricular and atrial arrhythmias were observed in the two DM1 cell lines. These findings enhance our comprehension of the molecular mechanisms underlying DM1 and provide valuable insights into the pathophysiology of ventricular and atrial involvement.

Generating atrial-like cardiomyocytes derived from human induced pluripotent stem cells (hiPSCs) is crucial for modeling and treating atrial-related diseases, such as atrial arrythmias including atrial fibrillations. However, it is essential to obtain a comprehensive understanding of the electrophysiological properties of these cells. The objective of the present study was to investigate the molecular, electrical, and biophysical properties of several ion channels, especially NaV1.5 channels, in atrial hiPSC cardiomyocytes. Atrial cardiomyocytes were obtained by the differentiation of hiPSCs treated with retinoic acid (RA). The quality of the atrial specification was assessed by qPCR, immunocytofluorescence, and western blotting. The electrophysiological properties of action potentials (APs), Ca2+ dynamics, K+ and Na+ currents were investigated using patch-clamp and optical mapping approaches. We evaluated mRNA transcript and protein expressions to show that atrial cardiomyocytes ex...

Background: The ability to differentiate patient-specific human induced pluripotent stem cells (hiPSC) into cardiac myocytes (hiPSC-CM) offers novel perspectives for cardiovascular research. A number of studies, which reported mainly on current-voltage curves used hiPSC-CM to model voltage-gated Na þ channel (Na v ) dysfunction. However, the expression patterns and precise biophysical and pharmacological properties of Na v channels from hiPSC-CM remain unknown. Our objective was to study the characteristics of Na v channels from hiPSC-CM and assess the appropriateness of this novel cell model. Methods: We generated hiPSC-CM using the recently described monolayer-based differentiation protocol. Results: hiPSC-CM expressed cardiac-specific markers, exhibited spontaneous electrical and contractile activities, and expressed distinct Na v channels subtypes. Electrophysiological, pharmacological, and molecular characterizations revealed that, in addition to the main Na v 1.5 channel, the neuronal TTX-sensitive Na v 1.7 channel was also significantly expressed in hiPSC-CM. Most of the Na þ currents were resistant to tetrodotoxin block. Therapeutic concentrations of lidocaine, a class I antiarrhythmic drug, also inhibited Na þ currents in a use-dependent manner. Na v 1.5 and Na v 1.7 expression and maturation patterns of hiPSC-CM and native Human cardiac tissues appeared to be similar. The four Na v b regulatory subunits were expressed in hiPSC-CM, with b3 being the preponderant subtype. Conclusions: The findings indicated that hiPSC-CM robustly express Na v 1.5 channels which exhibited molecular and pharmacological properties similar to those in native cardiac tissues. Interestingly, neuronal Na v 1.7 channels were also expressed in hiPSC-CM and are expected to be responsible for the TTX-sensitive Na v current.

Voltage gated sodium channels (Na v channels) play an important role in nociceptive transmission. They are intimately tied to the genesis and transmission of neuronal firing. Five different isoforms (Na v 1.3, Na v 1.6, Na v 1.7, Na v 1.8, and Na v 1.9) have been linked to nociceptive responses. A change in the biophysical properties of these channels or in their expression levels occurs in different pathological pain states. However, the precise involvement of the isoforms in the genesis and transmission of nociceptive responses is unknown. The aim of the present study was to investigate the synergy between the different populations of Na v channels that give individual neurons a unique electrophysical profile. We used the patch-clamp technique in the whole-cell configuration to record Na v currents and action potentials from acutely dissociated small diameter DRG neurons (<30 μm) from adult rats. We also performed single cell qPCR on the same neurons. Our results revealed that there is a strong correlation between Na v currents and mRNA transcripts in individual neurons. A cluster analysis showed that subgroups formed by Na v channel transcripts by mRNA quantification have different biophysical properties. In addition, the firing frequency of the neurons was not affected by the relative populations of Na v channel. The synergy between populations of Na v channel in individual small diameter DRG neurons gives each neuron a unique electrophysiological profile. The Na v channel remodeling that occurs in different pathological pain states may be responsible for the sensitization of the neurons.

Background and Aim-The action potential plateau of Purkinje fibers is particularly sensitive to tetrodotoxin (TTX) and this could be due to a TXX-sensitive Na + current. The expression of TTXsensitive neuronal Na V 1.1 and Na V 1.2 isoforms has been reported in canine Purkinje myocytes. Our aim was to investigate by means of biochemical and functional techniques whether the TTX-sensitive skeletal Na V 1.4 isoform is also expressed in canine cardiac Purkinje myocytes. Methods and Results-Using Na V 1.4 specific primers, a PCR product corresponding to Na V 1.4 was amplified from canine Purkinje fibers RNA and confirmed by sequencing and megablast of the gene bank. Confocal indirect immunostaining using anti-Na V 1.4 antibody demonstrates distinct sarcolemmal staining pattern compared to that of the cardiac isoform Na V 1.5. Expression of Na V 1.4 in tsA201 cells yielded a TTX-sensitive Na + current with an IC 50 of 10 nM. Conclusions-These results demonstrate the expression of the TTX-sensitive Na V 1.4 channel in canine cardiac Purkinje myocytes. This novel finding suggests a role of Na V 1.4 channel in Purkinje myocytes and thus has important clinical implications for the mechanisms and management of ventricular arrhythmias originating in the Purkinje network.

Genome-wide association studies (GWAS) have identified thousands of genetic variants linked to the risk of human disease. However, GWAS have so far remained largely underpowered in relation to identifying associations in the rare and low-frequency allelic spectrum and have lacked the resolution to trace causal mechanisms to underlying genes 1 . Here we combined whole-exome sequencing in 392,814 UK Biobank participants with imputed genotypes from 260,405 FinnGen participants (653,219 total individuals) to conduct association meta-analyses for 744 disease endpoints across the protein-coding allelic frequency spectrum, bridging the gap between common and rare variant studies. We identified 975 associations, with more than one-third being previously unreported. We demonstrate population-level relevance for mutations previously ascribed to causing single-gene disorders, map GWAS associations to likely causal genes, explain disease mechanisms, and systematically relate disease associations to levels of 117 biomarkers and clinical-stage drug targets. Combining sequencing and genotyping in two population biobanks enabled us to benefit from increased power to detect and explain disease associations, validate findings through replication and propose medical actionability for rare genetic variants. Our study provides a compendium of protein-coding variant associations for future insights into disease biology and drug discovery. Inherited variations in protein-coding and non-coding DNA have a role in the risk, onset and progression of human disease. Traditionally, geneticists have dichotomized diseases as either caused by coding mutations in single genes that tend to be rare, highly penetrant and frequently compromise survival and reproduction (often termed 'Mendelian' diseases), or as common diseases that show a complex pattern of inheritance influenced by the joint contributions of hundreds of low-impact, typically non-coding genetic variants (often termed 'complex' diseases). For both rare and common conditions, large human cohorts systematically characterized for a respective trait of interest have enabled the identification of thousands of disease-relevant variants through either sequencing-based approaches or GWAS. Nevertheless, the exact causal alleles and mechanisms that underlie associations of genetic variants to disease have so far remained largely elusive 1 . In recent years, population biobanks have been added to the toolkit for disease gene discovery. Biobanks provide the opportunity to simultaneously investigate multiple traits and diseases at once and uncover relationships between previously unconnected phenotypes. For instance, the UK Biobank (UKB) is a resource that captures detailed phenotype information matched to genetic data for more than 500,000 individuals and, since its inception, has facilitated biomedical discoveries at an unprecedented scale 2 . We and others have recently reported on the ongoing efforts to sequence the exomes of all UKB participants and link genetic findings to a broad range of phenotypes . We also established FinnGen (FG) (), an academic-industry collaboration to identify genotype-phenotype correlations in the Finnish founder population with the aim of better understanding how the genome affects health. Finland is a well-established genetic isolate and a unique gene pool distinguishes Finns from other Europeans 7 . The distinct Finnish haplotype structure is characterized by large blocks of co-inherited DNA in linkage disequilibrium and an enrichment for alleles that are rare in other populations, but can still be confidently imputed from genotyping data even in the rare and ultra-rare allele frequency spectrum . Through combining imputed genotypes with detailed phenotypes ascertained through national registries, FG holds the promise to provide particular insights into the so far little examined allele frequency spectrum between 0.1 and 2%, where both sequencing studies and GWAS have so far remained largely underpowered in relation to identifying associations with disease. This spectrum includes many coding variants with moderate to large effect sizes that can help

Synaptic and axonal glutamatergic signaling to NG2 glia in white matter is critical for the cells' differentiation and activity dependent myelination. However, in gray matter the impact of neuron-to-NG2 glia signaling is still elusive, because most of these cells keep their non-myelinating phenotype throughout live. Early in postnatal development, hippocampal NG2 glia express AMPA receptors with a significant Ca 2+ permeability allowing for plasticity of the neuron-glia synapses, but whether this property changes by adulthood is not known. Moreover, it is unclear whether NG2 glia express auxiliary transmembrane AMPA receptor related proteins (TARPs), which modify AMPA receptor properties, including their Ca 2+ permeability. Through combined molecular and functional analyses, here we show that hippocampal NG2 glia abundantly express TARPs γ4, γ7, and γ8 as well as cornichon (CNIH)-2. TARP γ8 undergoes profound downregulation during development. Receptors of adult NG2 glia showed an increased sensitivity to blockers of Ca 2+ permeable AMPA receptors, but this increase mainly concerned receptors located close to the soma. Evoked synaptic currents of NG2 glia were also sensitive to blockers of Ca 2+ permeable AMPA receptors. The presence of AMPA receptors with varying Ca 2+ permeability during postnatal maturation may be important for the cells' ability to sense and respond to local glutamatergic activity and for regulating process motility, differentiation, and proliferation.

P2X receptors are ligand-gated ion channels which are activated by excitatory neurotransmitter ATP. Despite considerable evidence of signaling by extracellular nucleotides in other sensory systems, P2X receptors in the visual system have only rarely been studied, and almost nothing is known about their functional significance in the retina. To determine whether ATP plays a role in the modulation of vertical retinal signal pathways, we examined the expression of P2X receptor mRNA in freshly isolated bipolar cells of the rat retina Ž . Brown Norway, P25 using the single-cell RT-PCR technique. Positive amplification signals were found in about 33% of the bipolar cells for P2X , P2X and P2X but not for P2X mRNA. We conclude that at least a subpopulation of bipolar cells in the rat retina 3 4 5 7 expresses ionotropic P2 receptors of the P2X type and that these possibly exert a neuromodulatory influence on information processing in the retina.

Since animal models are inevitable for medical research, information on species differences in glial cell properties is critical for successful translational research. Here, we review current knowledge about morphological and functional properties of human astrocytes and NG2 glial cells and compare these data with those obtained for the comparable cells in rodents. Morphological analyses of astrocytes in the neocortex of rodents versus humans have demonstrated clear differences. In contrast, the functional properties of astrocytes or NG2 glial cells in these species are surprisingly similar. However, these findings should be interpreted with caution, as so far functional analyses of human cells are only available from neocortex and hippocampus, and it is known from rodent studies that the properties of astrocytes in different brain regions may vary considerably. Moreover, technical challenges render astrocyte electrophysiological measurements in situ unreliable, and human cell properties may be affected by medications. Nevertheless, based on the limited data currently available, there is substantial similarity between human and rodent astrocytes with regard to those functional properties studied to date. The unique morphological characteristics of astrocytes in human neocortex call for further physiological analysis. The basic properties for NG2 glia are even less completely evaluated with regard to the question of species differences but no glaring differences have been reported so far. In conclusion, it remains justifiable to employ mouse or rat models to investigate the etiology of human CNS diseases that might involve astrocytes or NG2 glia.

NG2 cells are equipped with transmitter receptors and receive direct synaptic input from glutamatergic and GABAergic neurons. The functional impact of these neuron-glia synapses is still unclear. Here, we combined functional and molecular techniques to analyze properties of GABA A receptors in NG2 cells of the juvenile mouse hippocampus. GABA activated slowly desensitizing responses in NG2 cells, which were mimicked by muscimol and inhibited by bicuculline. To elucidate the subunit composition of the receptors we tested its pharmacological properties. Coapplication of pentobarbital, benzodiazepines, and zolpidem all significantly increased the GABA-evoked responses. The presence of small tonic currents indicated the presence of extrasynaptic GABA A receptors. To further analyze the subunit expression, single cell transcript analysis was performed subsequent to functional characterization of NG2 cells. The subunits ␣1, ␣2, ␤3, ␥1, and ␥2 were most abundantly expressed, matching properties resulting from pharmacological characterization. Importantly, lack of the ␥2-subunit conferred a high Zn 2ϩ sensitivity to the GABA A receptors of NG2 cells. Judging from the zolpidem sensitivity, postsynaptic GABA A receptors in NG2 cells contain the ␥2-subunit, in contrast to extrasynaptic receptors, which were not modulated by zolpidem. To determine the effect of GABA A receptor activation on membrane potential, perforated patch recordings were obtained from NG2 cells. In the current-clamp mode, GABA depolarized the cells to approximately Ϫ30 mV, indicating a higher intracellular Cl Ϫ concentration (ϳ50 mM) than previously reported. GABA-induced depolarization in NG2 cells might trigger Ca 2ϩ influx through voltage-activated Ca 2ϩ channels.

P2X receptors are ligand-gated ion channels activated by adenosine triphosphate and expressed in a broad variety of tissues. The present study demonstrates the expression of various types of purinergic P2X receptors in identified retinal ganglion cells (RGCs) of the adult rat retina. Single-cell reverse transcription polymerase chain reaction (SC-RT-PCR) resulted in a positive amplification signal for all P2X receptor subunit mRNAs examined (P2X , P2X ). Immunohistochemistry with P2X receptor subunit-specific antibodies showed 3 -5 7 3,4 a labelling of neurons in the ganglion cell layer and inner nuclear layer. Our data suggest that extracellular ATP acts directly on RGCs via several types of P2X receptors and may provide neuromodulatory influences on information processing in the retina.

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P2X receptors are ligand-gated ion channels activated by ATP. They are expressed in a broad variety of tissues. To date, eight P2X Ž . receptor subunits P2X -P2X , P2XM have been cloned. In spite of the considerable evidence of signaling by extracellular nucleotides 1 7 in other sensory systems, only few studies have been undertaken in the retina. In earlier studies, we have demonstrated that there is mRNA expression of the P2X and P2X subunits in the rat retina. In the present study, molecular biological methods were used to 2 -5 7 Ž . Ž . investigate expression of P2X receptor mRNA in freshly isolated Muller cells MCs of the adult rat retina Brown Norway . A total of 36 MCs was analyzed, employing the single-cell RT-PCR. A positive amplification signal of 11r14 for P2X -mRNA, 5r10 for 3 P2X -mRNA, 3r10 for P2X -mRNA and 0r8 for P2X -mRNA was revealed. Additionally, the astroglial identity of the cells under 4 5 7 Ž . studied was confirmed in 10 cases by simultaneous amplification of RT-PCR products of glutamine synthetase GS -and P2X-mRNA. We conclude that MCs of rat retina express ionotropic P2 receptors, which, in addition to other functions, may play a key role within the recently described long range calcium signaling and the fast direct glia-neuron interactions in the rat retina.

NG2 glia receive synaptic input from neurons, but the functional impact of this glial innervation is not well understood. In the developing cerebellum and somatosensory cortex the GABAergic input might regulate NG2 glia differentiation and myelination, and a switch from synaptic to extrasynaptic neuron–glia signaling was reported in the latter region. Myelination in the hippocampus is sparse, and most NG2 glia retain their phenotype throughout adulthood, raising the question of the properties and function of neuron-NG2 glia synapses in that brain region. Here, we compared spontaneous and evoked GABAA receptor-mediated currents of NG2 glia in juvenile and adult hippocampi of mice of either sex and assessed the mode of interneuron–glial signaling changes during development. With patch-clamp and pharmacological analyses, we found a decrease in innervation of hippocampal NG2 glia between postnatal days 10 and 60. At the adult stage, enhanced activation of extrasynaptic receptors occurre...

Extracellular purines are important signaling molecules involved in numerous physiological and pathological processes via the activation of P2 receptors. Information about the spatial and temporal P2 receptor (P2R) expression and its regulation remains crucial for the understanding of the role of P2Rs in health and disease. To identify cells carrying P2X2Rs in situ, we have generated BAC transgenic mice that express the P2X2R subunits as fluorescent fusion protein (P2X2-TagRFP). In addition, we generated a BAC P2Y1R TagRFP reporter mouse expressing a TagRFP reporter for the P2RY1 gene expression. We demonstrate expression of the P2X2R in a subset of DRG neurons, the brain stem, the hippocampus, as well as on Purkinje neurons of the cerebellum. However, the weak fluorescence intensity in our P2X2R-TagRFP mouse precluded tracking of living cells. Our P2Y1R reporter mice confirmed the widespread expression of the P2RY1 gene in the CNS and indicate for the first time P2RY1 gene expressi...

Current approaches to monitor and quantify cell division in live cells, and reliably distinguish between acytokinesis and endoreduplication, are limited and complicate determination of stem cell pool identities. Here we overcome these limitations by generating an in vivo reporter system using the scaffolding protein anillin fused to enhanced green fluorescent protein, to provide high spatiotemporal resolution of mitotic phase. This approach visualizes cytokinesis and midbody formation as hallmarks of expansion of stem and somatic cells, and enables distinction from cell cycle variations. High-resolution microscopy in embryonic heart and brain tissues of enhanced green fluorescent protein-anillin transgenic mice allows live monitoring of cell division and quantitation of cell cycle kinetics. Analysis of cell division in hearts post injury shows that border zone cardiomyocytes in the infarct respond with increasing ploidy, but not cell division. Thus, the enhanced green fluorescent protein-anillin system enables monitoring and measurement of cell division in vivo and markedly simplifies in vitro analysis in fixed cells.

Equisetum hyemale species is considered a medicinal plant used in the form of infusions to combat infectious or inflammation diseases and also diuretic effects, presenting several compounds related to these actions. In previous studies different species of Equisetum showed several phenolic compounds. The objective of this study was, for the first time, based on phytochemistry analysis to evaluate the antioxidant and antimicrobial activity. The 70% ethanolic and methanolic extracts of E. hyemale were characterized by spectrophotometric and high-performance liquid chromatography with pulsed amperometric detector analyses, as well as its antioxidant potential based on the scavenger activity of 2,2-diphenyl-1-picrylhydrazyl (DPPH). In addition was verified the antimicrobial activity by broth microdilution technique against bacteria and fungi. The extracts showed phytochemical similarity, which demonstrated the presence of phenolic compounds, the scavenging activity for free radicals was about 30% and was observed better antifungal activity against dermatophyte fungi, with minimum inhibitory concentration and minimum fungicidal concentration of 0.62 mg/mL to Trichophyton rubrum and Microsporum canis. The extracts exhibits great potential to therapeutic applications or product development, since both possess antifungal activity and antioxidant action associated with little difference in their phytochemical composition.

The mating behaviors of crickets, especially those related to agonistic encounters and oviposition, are poorly known. For example, only 10 of the 1005 valid species of Phalangopsidae have been studied to some extent. Here, we describe the reproductive behavior of Endecous (Endecous) chape, characterizing the actions involved in agonistic encounters, mating behaviors (female attraction, pair-formation, courtship, mating, and postcopulatory behavior), and oviposition. We recorded and timed agonistic, mating, and oviposition behaviors in staged trials. The male-male interactions of E. chape ranged in aggressiveness from low intensity (only antennal interaction) to high intensity (reciprocal fights). In the mating behavior, males courted females through antennation of the females' abdomen and cerci, followed by production of acoustic signals (with the exception of two males). Copulation occurred with females positioned above males (as is typical of Phalangopsidae), with an average elapsed time of 684.13 s, which is shorter than in other Phalangopsidae. We observed oviposition behavior only when three gravid females were placed together in an arena. Here, we contribute new knowledge of phalangopsid cricket behavior and provide useful information for understanding the evolution of reproductive behaviors. New characters described here can be used in phylogenetic analysis and for future studies about sexual selection and natural history.

Pair-formation, courtship, copulatory, and post-copulatory behaviors of Eidmanacris meridionalis Desutter-Grandcolas, 1995, were described under laboratory conditions and compared with congeneric species and other Phalangopsidae crickets whose mating behavior has been previously studied. Field observations for the species are also reported. Mating behaviors were observed in E. meridionalis couples: they first remained motionless, and then began walking through the arena screening the substrate. Mating behavior started with antennal contact, followed by males positioning themselves in front of females, that, when receptive, mounted on the males back and started feeding on metanotum secretions of the males. Copulation (female-above-male position) started with the engaging of copulatory structures and stopped with couple detachment. The end-to-end position was observed once, right after couple detachment. Males always remained with the spermatophore, which was eaten after removal or, alternatively, left in the substrate. The main differences in mating behavior of Eidmanacris species regard the mating position and the duration of the reproductive stages, suggesting that other reproductive repertoires can be observed in the genus since 27 species have not yet been studied in regards their reproductive behavior. We can also assume that the morphological, genetic, and chemical diversity found among the species of Phalangopsidae reflects in the behaviors of pair-formation, courtship, copulation, and post-copulation, leading to a diversity of copulation positions, duration of the stages, methods of attracting partners and parental investment. The description of Phalangopsidae mating patterns can provide important information for future evolutionary and phylogenetic studies, apart from useful for distinguishing cryptic species.

The mating behaviors of crickets, especially those related to agonistic encounters and oviposition, are poorly known. For example, only 10 of the 1005 valid species of Phalangopsidae have been studied to some extent. Here, we describe the reproductive behavior of Endecous (Endecous) chape, characterizing the actions involved in agonistic encounters, mating behaviors (female attraction, pair-formation, courtship, mating, and post-copulatory behavior), and oviposition. We recorded and timed agonistic, mating, and oviposition behaviors in staged trials. The male-male interactions ofE.chaperanged in aggressiveness from low intensity (only antennal interaction) to high intensity (reciprocal fights). In the mating behavior, males courted females through antennation of the females’ abdomen and cerci, followed by production of acoustic signals (with the exception of two males). Copulation occurred with females positioned above males (as is typical of Phalangopsidae), with an average elapsed...

O conhecimento do espalhamento e do tempo de evaporacao de gotas de pulverizacao e fundamental para entender a interacao entre a calda de fungicida e a superficie alvo para que aconteca adequada distribuicao do fungicida e protecao do alvo. O objetivo desta pesquisa foi determinar a area de molhamento e o tempo de evaporacao de gotas contendo o fungicida Cabrio® Top sem e com o adjuvante Nimbus® depositadas em superficies foliares das culturas de tomate, pimentao, pepino e superficie artificial (lâmina de vidro). Foi utilizado um sistema de analises por imagens composto de um gerador de gotas, um estereoscopio com câmera para captura de imagens e uma câmara climatica de controle de temperatura e umidade relativa do ar. Gotas de 600 µm de diâmetro contendo as caldas de pulverizacao foram depositadas nas superficies foliares e lâmina de vidro e as imagens sequenciais foram usadas para quantificar a area de molhamento e o tempo de evaporacao. A calda de pulverizacao e a superficie alvo...

Quantification of the effects of adjuvants on droplet behaviour on plant surfaces is needed to improve pesticide spray application efficiency for soybeans. Dispersion and evaporation of single 300-mm diameter droplets amended with each of four spray adjuvants at five concentrations were investigated for four soybean plant surfaces (abaxial and adaxial leaflet surfaces, petiole, basal stem). The four adjuvants were a crop oil concentrate (COC), a modified seed oil (MSO), a non-ionic surfactant (NIS) and an oil surfactant blend (OSB). A single-droplet generator was used to produce and deposit 300-mm diameter droplets on target surfaces under controlled environmental conditions. Adjuvants significantly increased the dispersion (or wetted area) of droplets on plant surfaces. Droplet-wetted areas increased with increased adjuvant concentrations but not in direct proportion. The average increases of wetted areas across the four soybean plant surfaces were 443, 462, 416, or 343% when the spray mixture was amended with COC, MSO, NIS or OSB at the manufacturer-recommended concentrations, respectively. Among the four surfaces, the largest wetted area was on the abaxial surface, followed by the adaxial surface, the petiole and then the basal stem. Droplet evaporation times were inversely proportional to the wetted areas. The evaporation time of 300-mm diameter droplets ranged from 36 to 142 s on the four surfaces when the spray mixture was amended with an adjuvant, whereas the water-only droplets ranged from 161 to 190 s. The results demonstrated that use of adjuvants offers great potential to improve the homogeneity of sprayed pesticides, to increase spray coverage and to reduce pesticide application rates on soybean plants. These effects could benefit farmers economically and reduce environmental contamination by pesticides.

In this study, we considered progression of liver diseases. Particularly we considered Hepatocellular Carcinoma Cancer, HCC, whose patients have low survival rates. For this purpose, we researched molecular structures and protein interactions involved in the initiation and progression of HCC. We exploited microarray data samples and their gene expression profiles from literature. During analysis, we implemented statistical data analysis techniques and looked for Differentially Expressed Genes during the initiation and progression of HCC. As a result of this analysis we found 12 hub genes, where 3 of them (ANLN, TOP2A and ASPM) were upregulated and the others (CXCL14, LINC01093, OIT3, CLEC4G, THRSP, APOF, CLTRN and FCN3) were downregulated. By performing Gene Ontology Analysis, we classified genes with increased or decreased expressions in terms of cellular component, biological process, and molecular function. Subsequently, we executed protein-protein interaction network analysis an...

The aim of this experiment was to determine the effects of dietary sepiolite supplementation on performance, carcass characteristics, blood parameters and rumen fluid metabolites in Merino cross breed lambs. For this purpose, a total of 21 weaned lambs, which were 3-3.5 months of age were allocated to one control group and two treatment groups each containing 7 lambs. The experiment lasted for a total of 90 days, of which the adaptation and experimental periods were 22 and 68 days, respectively. Pelleted alfalfa and concentrates were given to lambs individually. Concentrate diets of first and the second treatment groups were supplemented with 1 and 2% sepiolite as top dressed, respectively. There were no significant differences observed in body weights, body weight gains, hot and cold carcass yields, meat pH and meat composition among the groups. Relative weights of internal organs and carcass parts were also not affected by sepiolite supplementation. The addition of 1 and 2% sepiolite to the diets of animals increased the consumption of concentrates and total dry matter intake (P < .05). There were no differences among the groups in terms of consumption of dry matter of alfalfa, concentrates and total feed required for one kg body weight gain in lambs. However, body weight gain was increased by 4.22% and total feed conversion ratio was improved by 4.56% with 2% dietary sepiolite supplementation. Haematological and biochemical blood parameters and fecal dry matter were also not affected by the usage of sepiolite. Blood serum IgG levels were increased with sepiolite supplementation (P < .05). While the percentage of acetic acid in the total volatile fatty acids (VFA) in rumen fluid was decreased, neither the proportion of other volatile fatty acids nor the quantity of total volatile fatty acids were affected by dietary addition of sepiolite. It was concluded that 2% sepiolite supplementation in diets of Merinos cross bred lambs as top dressed may have economic benefits based on animal's immunity enhancement and some improvement in body weight gain and total feed conversion ratio without any negative effects on performance, carcass characteristics, rumen metabolites and blood parameters.

The genus Charaea Baly is distributed in the eastern Palaearctic, Himalayas, China and adjacent countries of the Oriental Region. Currently, 59 species of the genus Charaea have been recorded. The species of Charaea is characterised with a robust tubular aedeagus that terminates with a more or less distinct apical process with the Charaea coomani group having an internal sac with long sharp lateral sclerites. Up to now, 13 species of this group have been described in the Oriental Region, four of which are found in Vietnam. Charaea dinhcuongisp. nov. is described as a new species, based on specimens collected from Phu Quoc Island in southern Vietnam. Colour photographs of habitus and body details and DNA barcode sequences are presented. An identification key is provided for all Vietnamese species from the Charaea coomani group.

Studies in model organisms have contributed to elucidate multiple levels at which regulation of eukaryotic DNA replication occurs. Cdc7, an evolutionarily conserved serine±threonine kinase, plays a pivotal role in linking cell cycle regulation to genome duplication, being essential for the ®ring of DNA replication origins. Binding of the cell cycle-regulated subunit Dbf4 to Cdc7 is necessary for in vitro kinase activity. This binding is also thought to be the key regulatory event that controls Cdc7 activity in cells. Here, we describe a novel human protein, Drf1, related to both human and yeast Dbf4. Drf1 is a nuclear cell cycleregulated protein, it binds to Cdc7 and activates the kinase. Therefore, human Cdc7, like cyclin-dependent kinases, can be activated by alternative regulatory subunits. Since the Drf1 gene is either absent or not yet identi®ed in the genome of model organisms such as yeast and Drosophila, these ®ndings introduce a new level of complexity in the regulation of DNA replication of the human genome.

Pangolins are the most trafficked mammal in the world, and all eight species are listed under CITES Appendix I. DNAbased wildlife forensic techniques are recognized as an important component of investigating a pangolin seizure. In particular, determining the species of pangolin in a seizure will 1) confirm the presence of pangolin to establish the legality of any trade, and 2) ensure appropriate laws are applied to their fullest extent in a prosecution. Furthermore, valuable intelligence data, such as determining the geographic provenance of samples, can be produced through analysis of pangolin seizures. Despite the immense scale of the pangolin trade, standardized wildlife forensic techniques for testing pangolin seizures are in their infancy. To address this, here, we present a standardized genetic marker suitable for species identification of all eight pangolin species, and outline practical strategies for sampling large-volume pangolin scale seizures. We assessed the repeatability, reproducibility, robustness, sensitivity and phylogenetic resolution of this species identification test. Critically, the assay was tested in four wildlife forensic laboratories involved in testing pangolins. Additionally, we demonstrated the test's utility to conduct geographic provenance analysis of Phataginus tricuspis samples. We analysed five large-volume pangolin scale seizures in Malaysia, which elucidated key target species, poaching hotspots, and trafficking routes. Phataginus tricuspis was the most commonly identified species (88.8%) from the seizure samples, and 84.3% of these P. tricuspis individuals were likely sourced from western central Africa. We expect the implementation of the techniques presented in this paper will improve enforcement of pangolin trafficking crimes.

Tylosin has been used as a livestock feed additive and antibiotic growth promoter for many years. However, the mode of action by which tylosin enhances animal growth is unclear. We used high throughput sequencing of 16S rRNA genes to investigate the effects of tylosin as a feed additive on swine gut microbiota. No significant difference in the rate of weight increase was observed between control and tylosin-treated pigs during a 10 week feeding trial. However, tylosin-treated pigs showed rapid increases in the relative abundance of the phylum Firmicutes. Increases in Firmicutes species are associated with (so-called) obese type gut microbiota. The abundance of species of four families of the phylum Firmicutes (Streptococcaceae, Peptococcaceae, Peptostreptococcaceae, and Clostridiaceae) correlated positively with host weight gain. The abundance of Streptococcaceae family bacteria was least affected by tylosin-treatment. Distribution analysis of operational taxonomic units (OTUs) show...

Brain-computer interfaces (BCIs) have shown promising results in restoring motor function to individuals with spinal cord injury. These systems have traditionally focused on the restoration of upper extremity function; however, the lower extremities have received relatively little attention. Early feasibility studies used noninvasive electroencephalogram (EEG)-based BCIs to restore walking function to people with paraplegia. However, the limited spatiotemporal resolution of EEG signals restricted the application of these BCIs to elementary gait tasks, such as the initiation and termination of walking. To restore more complex gait functions, BCIs must accurately decode additional degrees of freedom from brain signals. In this study, we used subdurally recorded electrocorticogram (ECoG) signals from able-bodied subjects to design a decoder capable of predicting the walking state and step rate information. We recorded ECoG signals from the motor cortices of two individuals as they walk...

Background: Clinical treatments with typical antipsychotic drugs (APDs) are accompanied by extrapyramidal motor sideeffects (EPS) such as hypokinesia and catalepsy. As little is known about electrophysiological substrates of such motor disturbances, we investigated the effects of a typical APD, a-flupentixol, on the motor behavior and the neuronal activity of the whole basal ganglia nuclei in the rat. The motor behavior was examined by the open field actimeter and the neuronal activity of basal ganglia nuclei was investigated using extracellular single unit recordings on urethane anesthetized rats. We show that aflupentixol induced EPS paralleled by a decrease in the firing rate and a disorganization of the firing pattern in both substantia nigra pars reticulata (SNr) and subthalamic nucleus (STN). Furthermore, a-flupentixol induced an increase in the firing rate of globus pallidus (GP) neurons. In the striatum, we recorded two populations of medium spiny neurons (MSNs) after their antidromic identification. At basal level, both striato-pallidal and striato-nigral MSNs were found to be unaffected by a-flupentixol. However, during electrical cortico-striatal activation only striato-pallidal, but not striato-nigral, MSNs were found to be inhibited by a-flupentixol. Together, our results suggest that the changes in STN and SNr neuronal activity are a consequence of increased neuronal activity of globus pallidus (GP). Indeed, after selective GP lesion, a-flupentixol failed to induce EPS and to alter STN neuronal activity. Our study reports strong evidence to show that hypokinesia and catalepsy induced by a-flupentixol are triggered by dramatic changes occurring in basal ganglia network. We provide new insight into the key role of GP in the pathophysiology of APD-induced EPS suggesting that the GP can be considered as a potential target for the treatment of EPS.

Introduction : Le Covid-19 est la première pandémie du 21ème siècle. Causée par un virus émergent, le SARS-CoV-2, son diagnostic de confirmation actuel est basé sur la RT-PCR. La présente revue systématique avait comme objectif de décrire la performance des différents tests antigéniques et sérologiques dans le diagnostic du COVID-19 par rapport au Gold standard. Méthodes : La recherche a été réalisée sur les bases de données PubMed et Scopus, jusqu'au 26/04/2020. Ont été inclues les études ayant comparé les performances des tests sérologiques et/ou antigéniques (Sensibilité (Se) et Spécificité (Sp), valeur prédictive positive (VPP) et valeur prédictive négative (VPN), efficacité diagnostique) avec le test de référence RT-PCR dans le diagnostic du Covid-19. Résultats : Treize études remplissaient les critères de recherche sur un total de 2247 participants. Sept études ont été menées en Asie et six en Europe. Ont été identifiés 25 tests diagnostiques dont 22 étaient sérologiques et 3 antigéniques. Leur Se variait entre 18,4 et 100% , tandis que leur Sp variait entre 14,2 et 100%. Leur efficacité diagnostique variait quant à elle entre 66,5 et 99%. Le test de Hoffman et al. basé sur la détection des anticorps anti-Sars-Cov2 de type IgG avait la meilleure sensibilité et spécificité avec des pourcentages respectifs de 93,1% et 99,2%. Conclusion : L'amélioration des performances de ces tests permettrait d'éclairer les stratégies de riposte futures en matière de diagnostic, de prise en charge et de surveillance face à cette urgence sanitaire Mots-clés Covid-19, performance diagnostique, tests sérologiques, tests antigéniques, tests rapides SUMMARY: Background: Covid-19 is the first pandemic of the 21st century. It caused by an emerging virus SARS-CoV-2. Diagnosis of suspected cases is confirmed by nucleic acid assays with real-time PCR (RT-PCR). The objective of this systematic review was to describe the performance of different antigenic and serological tests in the diagnosis of Covid-19 compared to the Gold standard. Methods: The search was performed on the PubMed and Scopus databases up to 26/04/2020. Only studies are comparing the diagnostic performance of serological and/or antigenic tests (Sensitivity (Se) and Specificity (Sp), Positive Predictive Value (PPV) and Negative Predictive Value (NPV), diagnostic efficiency) with the reference test RT-PCR in the diagnosis of Covid-19 were included. Results: Thirteen studies met the search criteria out of a total of 2247 participants. Seven studies were conducted in Asia and six in Europe. Twenty-five diagnostic tests were identified, 22 of which were serological and 3 antigenic. Their Se ranged from 18.4 to 100%, while their Sp ranged from 14.2 to 100%. Their diagnostic efficacy ranged from 66.5 to 99%. The test by Hoffman et al. based on the detection of anti-Sars-Cov2 antibodies of the IgG type had the best sensitivity and specificity with percentages of 93.1% and 99.2% respectively. Improving the performance of these tests would inform future response strategies for diagnosis, management and surveillance in the face of this health emergency.

Background Congenital digestive malformations are a leading cause of infant mortality and morbidity. There are currently no reliable estimates of the number of children born with these malformations and the diversity of the underlying causes requires a range of preventive approaches. Thus, to estimate the prevalence and mortality of these pathologies and to determine the interest of prenatal diagnosis in Casablanca, we initiated this study. Methods This was a cross-sectional study conducted at the Ibn Rochd University Hospital in Casablanca from January 1, 2016, to December 31, 2019, among newborns with congenital digestive malformations diagnosed by a specialist in visceral and digestive surgery. Data related to pregnancy, delivery, and time of diagnosis were collected through the medical record and complemented by clinical and surgical data from the hospitalization registry. In addition to the descriptive analysis of the results, the statistical analysis included Log Rank tests an...

Background Improving the quality of life of patients is a key concern in public health. In the context of improving therapeutic compliance, quality of life of patients undergoing anti-tuberculosis treatment, and prevention of antibiotic resistance, we conducted this systematic review to support risk minimization actions. Methods A literature review of risk management strategies targeting adverse drug reactions to anti-tuberculosis drugs was conducted, including published observational studies. Two independent reviewers identified studies through Medline, Scopus, and Web of Science. The methodological quality of the included studies was assessed using criteria defined by the Newcastle Ottawa Scale. Results Seven studies were included in this review, which identified four risk management strategies (psychological intervention, reduction of drug dose with or without prescription of adjunctive medications, change of medication, permanent or temporary discontinuation of medication). The ...

Hotel industry, as a very dynamic activity within the tourism industry, applies innovations in business and develops voluntary eco-business standards for developing sustainable tourism. The paper aims to assess business excellence of hotels that are holders of the international eco-certificate Green Key in Serbia, a country in the Western Balkan region, by using the BEX model. The study reviews and presents the current situation when it comes to implementation and valorisation of eco-principles and standards in hotel business in Serbia, while examining their business excellence, as well as the opportunities for better positioning in the international tourism market. The research results show that the examined companies do not have poor ranks of business excellence. It is recommended for these hotels to continue with the current business while implementing innovations in sustainable business in order to improve business results. By monitoring the value of the BEX index, it is possible to avoid business risks, while expanding eco-awareness and implementing sustainable business policies, which would help hotel companies improve their business.

1 diabetes is still unknown. Here we show that the activating receptor NKp46 recognizes mouse and human ligands on pancreatic beta cells. NK cells appeared in the pancreas when insulitis progressed to type 1 diabetes, and NKp46 engagement by beta cells led to degranulation of NK cells. NKp46-deficient mice had less development of type 1 diabetes induced by injection of a low dose of streptozotocin. Injection of soluble NKp46 proteins into nonobese diabetic mice during the early phase of insulitis and the prediabetic stage prevented the development of type 1 diabetes. Our findings demonstrate that NKp46 is essential for the development of type 1 diabetes and highlight potential new therapeutic modalities for this disease.

Frenk inciri (Opuntia ficus-indica), güney bölgelerimizde doğal olarak yetişen, kaktüsgiller ailesinden bir meyvedir. Bu çalışmada, Adana ilinin Bucak ve Bozcalar köyleri ve Mersin ilinin Yanıkkışla, İncirgediği, Kırıt, Cırbıklar, Taşobası, Kumdere, Çokak ve Aladağ köylerinden oluşan bölgede doğal olarak yetişen frenk inciri popülasyonu incelenmiştir. Sörvey çalışması ile iri meyveli ve nispeten az dikenli olduğu değerlendirilen genotipler çalışmaya dahil edilmiştir. Seçilen genotiplerde bazı temel meyve özellikleri çalışılmıştır. Temel bileşen ve kümeleme analizi ile ümitvar genotipler belirlenmiştir. Genotiplerde meyve ağırlıklarının 67.40-138.00g, yenilebilir meyve ağırlıklarının 25.70-82.77g ve SÇKM değerlerinin %9.10-13.23 arasında olduğu tespit edilmiştir. Kümeleme analizi sonucunda beş genotip ümitvar olarak öne çıkmıştır. Çalışma sonuçları Adana-Mersin il sınırındaki köylerde yetişen frenk inciri genotiplerinin meyve boyutları bakımından düşük varyasyon gösterirken, özellikl...

Carotid body glomus cells mediate essential reflex responses to arterial blood hypoxia. They are dopaminergic and secrete growth factors that support dopaminergic neurons, making the carotid body a potential source of patient-specific cells for Parkinson&#39;s disease therapy. Like adrenal chromaffin cells, which are also hypoxia-sensitive, glomus cells are neural crest-derived and require the transcription factors Ascl1 and Phox2b; otherwise, their development is little understood at the molecular level. Here, analysis in chicken and mouse reveals further striking molecular parallels, though also some differences, between glomus and adrenal chromaffin cell development. Moreover, histology has long suggested that glomus cell precursors are &#39;émigrés&#39; from neighbouring ganglia/nerves, while multipotent nerve-associated glial cells are now known to make a significant contribution to the adrenal chromaffin cell population in the mouse. We present conditional genetic lineage-trac...

Objectives: To present a general overview of rice agronomic practices and transplanting operations by considering the interactions of soil, plant, and machine relationship in line with the System of Rice Intensification (SRI) cultivation practice. Methods: Some of the problems challenging Malaysian rice growers, as well as yield increase and total rice production in the last four decades, were first addressed and discussed. The trend in the world rice production between 1961 and 2014 was used to predict the production in 2020 and to show that Southeast Asian countries are expected to increase their production by 27.2%. Findings: A consistently increasing pattern from 3.1 tons/ha during 1981 to 4.1 tons/ha in 2014 was observed in the rice yield of Malaysia due to the advances in technology and improved farming operations coupled with integrated management and control of resources. Various literatures were reviewed and their findings of the best transplanting practices were summarized to discuss how SRI contributes to the production of higher rice yield with improved transplanting practices through a more effective root system. Our review shows that wider spacing, availability of solar radiation, medium temperature, soil aeration, and nutrient supply promote shorter Phyllochrons which increase the number of tillers in rice. In this regard, modification and development of a transplanter that complies with SRI specification require determination of optimum transplanting spacing, seed rate, and planting pattern to significantly improve yield. Improvement: It was concluded that for maximum yield, the SRI method in Malaysia should emphasize on planting of one seedling per hill with space of 0.25 m for optimum water consumption, nutrient and pest management.

Induction of cell apoptosis is one of the major host defense mechanisms through which macrophages control Mycobacterium tuberculosis (Mtb) infection. However, the mechanisms underlying macrophage apoptosis triggered by Mtb infection are still largely unknown. In this study, a microarray profiling survey revealed 14 miRNAs were down-regulated in CD14+ monocytes from active pulmonary tuberculosis patients, and only the reduction of miR-20a-5p could be reversed after successful anti-tuberculosis treatment. Validation of miR-20a-5p expression was confirmed using real time qPCR. Moreover, miR-20a-5p expression also decreased in differentiated THP-1 macrophages after mycobacterial infection in vitro. Functional assays through forced or inhibited expression of miR-20a-5p in THP-1 macrophages demonstrated that miR-20a-5p functioned as a negative regulator of mycobacterial-triggered apoptosis. Importantly, inhibition of miR-20a-5p expression resulted in more efficient mycobacterial clearance...

A rat brain cDNA encoding for a novel protein with agmatinase activity was cloned and functionally expressed. The protein was expressed as a histidine-tagged fusion product with a molecular weight of about 63 kDa. Agmatine hydrolysis was strictly dependent on Mn 2+ ; K m and k cat values were 2.5 § 0.2 mM and 0.8 § 0.2 s ¡1 , respectively. The product putrescine was a linear competitive inhibitor (K i D 5 § 0.5 mM). The substrate speciWcity, metal ion requirement and pH optimum (9.5) coincide with those reported for Escherichia coli agmatinase, the best characterized of the agmatinases. However, as indicated by the k cat /K m (320 M ¡1 s ¡1 ), the recombinant protein was about 290-fold less eYcient than the bacterial enzyme. The deduced amino sequence revealed great diVerences with all known agmatinases, thus excluding the protein from the arginase family. It was, however, highly identical (>85%) to the predicted sequences for fragments of hypothetical or unnamed LIM domain-containing proteins. As a suggestion, the agmatinase activity is adscribed to a protein with an active site that promiscuously catalyze a reaction other than the one it evolved to catalyze.

Sex determination is an important developmental event in the life cycle of all sexually reproducing plants. Recent studies of sex determination in many plant species, from ferns to maize, have been fruitful in identifying the diversity of genetic and epigenetic factors that are involved in determining the sex of the flower or individual. In those species amenable to genetic analysis, significant progress has been made toward identifying mutations that affect sex expression. By studying the interactions among these genes, pictures of how sex-determining signals are perceived to activate or repress male-or female-specific genes are emerging.

This study looked at the factors that facilitated the spread of Phytophthora ramorum and P. kernoviae at two locations in the west of Scotland. Spore traps, river baiting, bait plants, and soil sampling were used to both confirm the presence of, and measure the amount of, inoculum in the environment in order to quantify the relationship between inoculum levels and disease development. Phytophthora ramorum was detected in spore traps at high levels under a sporulating host, but also at sites where hosts were not present, leading to the conclusion that inocula in low-level spore traps were the result of soil splash. Rhododendron and Vaccinium bait plants were also infected with P. ramorum via soil splash at sites where there was no sporulating host present. Phytophthora kernoviae was only detected in spore traps where there was a sporulating host overhead. Water baiting confirmed the presence of P. ramorum in two streams in one of the gardens, but P. kernoviae was not detected using this method at the other garden despite the large-scale P. kernoviae infection there. Inoculum continued to be detected in soil in areas where infected hosts had been removed 2 years ago, confirming that both of these pathogens can survive in soil for a considerable period. Evidence of the movement of infested mulch during horticultural activity was found. These findings have clear implications for the control of disease spread within the garden setting.

Pancreatic ductal adenocarcinoma (PDAC) tumours carry multiple gene mutations and respond poorly to treatments. There is currently an unmet need for drug carriers that can deliver multiple gene cargoes to target high solid tumour burden like PDAC. Here, we report a dual targeted extracellular vesicle (dtEV) carrying high loads of therapeutic RNA that effectively suppresses large PDAC tumours in mice. The EV surface contains a CD64 protein that has a tissue targeting peptide and a humanized monoclonal antibody. Cells sequentially transfected with plasmid DNAs encoding for the RNA and protein of interest by Transwell®-based asymmetric cell electroporation release abundant targeted EVs with high RNA loading. Together with a low dose chemotherapy drug, Gemcitabine, dtEVs suppress large orthotopic PANC-1 and patient derived xenograft tumours and metastasis in mice and extended animal survival. Our work presents a clinically accessible and scalable way to produce abundant EVs for deliveri...

Background Cancer subtype switching, which involves unclear cancer cell origin, cell fate decision, and transdifferentiation of cells within a confined tumor microenvironment, remains a major problem in pancreatic cancer (PDA). Results By analyzing PDA subtypes in The Cancer Genome Atlas, we identified that epigenetic silencing of apoptosis-associated tyrosine kinase (AATK) inversely was correlated with mRNA expression and was enriched in the quasi-mesenchymal cancer subtype. By comparing early mouse pancreatic lesions, the non-invasive regions showed AATK co-expression in cells with acinar-to-ductal metaplasia, nuclear VAV1 localization, and cell cycle suppression; but the invasive lesions conversely revealed diminished AATK expression in those with poorly differentiated histology, cytosolic VAV1 localization, and co-expression of p63 and HNF1α. Transiently activated AATK initiates acinar differentiation into a ductal cell fate to establish apical-basal polarization in acinar-to-du...

Background: Chitinase 3-like-1 (CHI3L1) is a secretion glycoprotein associated with the immunosuppressive tumor microenvironment (TME). The secretory mode of CHI3L1 makes it a promising target for cancer treatment. We have previously reported that Rab37 small GTPase mediates secretion of IL-6 in macrophages to promote cancer progression, whereas the roles of Rab37 in the intracellular trafficking and exocytosis of CHI3L1 are unclear. Methods: We examined the concentration of CHI3L1 in the culture medium of splenocytes and bone marrow derived macrophages (BMDMs) from wild-type or Rab37 knockout mice, and macrophage or T cell lines expressing wild type, active GTP-bound or inactive GDP-bound Rab37. Vesicle isolation, total internal reflection fluorescence microscopy, and real-time confocal microscopy were conducted. We developed polyclonal neutralizing-CHI3L1 antibodies (nCHI3L1 Abs) to validate the therapeutic efficacy in orthotopic lung, pancreas and colon cancer allograft models. Multiplex fluorescence immunohistochemistry was performed to detect the protein level of Rab37 and CHI3L1, and localization of the tumor-infiltrating immune cells in allografts from mice or tumor specimens from cancer patients. Results: We demonstrate a novel secretion mode of CHI3L1 mediated by the small GTPase Rab37 in T cells and macrophages. Rab37 mediated CHI3L1 intracellular vesicle trafficking and exocytosis in a GTP-dependent manner, which is abolished in the splenocytes and BMDMs from Rab37 knockout mice and attenuated in macrophage or T cell lines expressing the inactive Rab37. The secreted CHI3L1 activated AKT, ß-catenin and NF-κB signal pathways in cancer cells and macrophages to foster a protumor TME characterized by activating M2 macrophages and increasing the population of regulatory T cells. Our developed nCHI3L1 Abs showed the dual properties of reducing tumor growth/metastases and eliciting an immunostimulatory TME in syngeneic orthotopic lung, pancreas and colon tumor models. Clinically, high plasma level or intratumoral expression of CHI3L1 correlated with poor survival in 161 lung cancer, 155 pancreatic cancer and 180 colon cancer patients. Conclusions: These results provide the first evidence that Rab37 mediates CHI3L1 secretion in immune cells and highlight nCHI3L1 Abs that can simultaneously target both cancer cells and tumor microenvironment.