Methanoculleus, a methanogen commonly detected in marine and estuarine environments. The high percentage (35%) of unassigned fungal taxa suggested the presence of a high degree of undiscovered diversity within a structured fungal community (including both yeast and filamentous life forms) and reinforce the hypothesis of a high degree of biological uniqueness of the habitat under study.Here, we present the results related to a new unique terrestrial ecosystem found in an englacial hypersaline brine found in Northern Victoria Land (Antarctica). Both the geochemistry and microbial  diversity revealed an unicity with respect to all the other known Antarctic brines and suggested a probable ancient origin mainly due a progressive cryoconcentration of seawater. The prokaryotic community presented some peculiarities, such as the occurrence of sequences of Patescibacteria (which can thrive in nutrient-limited water environments) or few Spirochaeta, and the presence of archaeal sequences of Methanomicrobia closely related to  Brines were also discovered in the deep subsurface in Canada, Finland, Germany and Sweden. In Antarctica, hypersaline brines were found at McMurdo Sound, within the permafrost of the Taylor Valley (e.g.3), as well as below ice-sealed Antarctic lakes (e.g.4) or in the Antarctic subglacial lakes (i.e.5). Several studies modeled the occurrence and the effects of the subglacial brines or detected their occurrence through indirect methods (e.g.8). Despite these efforts the subglacial aquatic systems remain poorly understood mainly due to the lack of direct sampling. Englacial brines that flow within glaciers9, ice sheets or ice shelves10 are even less known. The connections between englacial and subglacial brines and their origins are still debated although at least in Taylor Valley several studies (i.e.3) provided strong inputs to their comprehension. Different mechanisms regarding the origin of the hypersaline brines were proposed (e.g.11). Subglacial and endoglacial brines can be extremely important in regard to glacier dynamics (e.g.13) and even more for ecological interest (e.g.14). Microbial communities sharing subglacial environment have been studied in recent decades, especially in order to understand possible effects on the weathering of the underlying rock (e.g.17). Probably, the most studied case is the outflow brine of Blood Falls in Taylor Valleys (e.g.20) that revealed the presence of a thriving community of chemosynthetic bacteria whose 74% of clones and isolates shared high 16S rRNA gene sequence homology with phylotypes from marine systems.Interest in brines within cryoenvironments has increased after they have been found on Mars (e.g.29); accordingly, they have been used as comparison in this paper. The phylogenetic groups found in the different frozen lakes were quite different each other and exhibited different dominant groups: e.g. Bacteroidetes and Actinobacteria in East Bonney Lake or Proteobacteria (and Cyanobacteria26) in Lake Vanda27.The microbial communities of hypersaline brines observed below ice-sealed Antarctic lakes were surely more investigated  at the date of sampling (26 November 2019) and remained liquid and flowing also at the last check (7 December 2019) when the level of brines raised upward 2.25 m. In situ the brine appeared quite rich in gas bubbles, with a temperature of − 17.4 °C, slightly basic (from 7.4 and 7.6), hypersaline (NaCl concentration is about 198 g L−1) and with a low percentage of DO (12.9%). The geochemical characteristics of the two samples of the same brine (BC-1 and BC-2) are reported in Table −1), sodium (2.7 eq L−1), magnesium (1.1 eq L−1) and potassium (110 meq L−1) with values that are close to only a few other Antarctic brines, such as the East Lake Bonney (ELB)31 and lake Vida brines (VD)21 although BC brines slightly exceed in sodium and potassium with respect to ELB and Lake Vanda (LV). Differently, sulfates (60 meq L−1) and calcium (160–170 meq L−1) showed values that are in line with the majority of the other Antarctic brines (e.g.31). The chemical peculiarity of BC brines with respect to the other Antarctic ones was preliminarily evaluated though Principal Component Analysis (PCA), including the six major ion concentrations as variables . PC1 significantly correlates with the ionic strength of the brines . Therefore, PC1 can be interpreted as indicative of the ionic activity of the brine solution. ELB, BC and, to a lesser extent, VD brines have the more negative PC1 values. The PC2, instead, indicates the main characteristics of the less saline LV and Tarn Flat (TF) brines, characterized by a more significant input of Ca2+ and sulfates, respectively. Figure 4 of several Antarctic brines, where it is possible to observe that Cl/SO4 of BC brines is approximately 10 times higher than seawater (SW), but similar to ELB brines. Conversely, Mg/K ratio in BC is almost the same as the seawater and it is sensibly lower than the Englacial and Blood Falls brines that were the most similar to BC for this parameter.The brines were found at 9.1 m of depth within a valley glacier characterized by a glacial unconformity and an erosional channel at 3 m of depth in which thin alluvial sediments (3 cm thick) were found. These peculiar structures are described in Forte et al.2 or NH4+, while the overall contribution of inorganic nitrogen was due to NO3. Similarly, to what was found by Lyons et al.9, we suppose that this value is an oxidation artifact due to the storage before the analysis. Moreover, the determination of the total N with the independent analysis carried out with Elemental Analyzer, provided values similar to NO3. Therefore, we prefer to report the total N concentration without any chemical speciation.Another peculiarity of BC brines  revealed a total number of 1042 ASVs, which were then resolved in 25 prokaryotic phyla, with a resolution of 162 genera. Unclassified sequences were on average 6.9 and 12.9% at phylum and genus levels, respectively.Methanoculleus (87%). Eight bacterial phyla occurred at a percentage greater than 0.1%, namely Actinobacteria, Bacteroidetes, Cyanobacteria, Firmicutes, Patescibacteria, Planctomycetes, Proteobacteria and Verrucomicrobia. Among them, Proteobacteria, Bacteroidetes, Verrucomicrobia, Patescibacteria, and Actinobacteria exceeded 1%. In particular, Proteobacteria (average percentage 48.6%) were particularly abundant and mainly represented by Gammaproteobacteria (family Marinobacteraceae) and, to a lesser extent, Alphaproteobacteria. The second most abundant group was that of Bacteroidetes , followed by Verrucomicrobia, Patescibacteria and Actinobacteria . Firmicutes, Planctomycetes and Cyanobacteria were in the range 0.3–0.7%.Archaea were present in a low percentage (0.5%) almost entirely affiliated to the genus Marinobacter (among Proteobacteria) was predominant (38.5%), followed at a lesser extent by Psychroflexus and Flavimarina , Psychrobacter , and Luteolibacter . Other genera occurring at a percentage between 1 and 5% were Gillisia, Salegentibacter, Aurantivirga, Mesonia and Algoriphagus, all among Bacteroidetes (63.8% of Bacteroidetes sequences were resolved at genus level). Ten genera were found among Actinobacteria, but none of them exceeded 1%. No genus was determined among Patescibacteria , Phoma (9%), Penicillium (8%), Phaeoisaria (4%), Aspergillus (3%), and to a lesser extent Cyphellophora (1.5%), Periconia (1.2%), Parengyodontium (1%), and Mucor (1%). Considering yeast life forms, Glaciozyma was the unique yeast genus exhibiting an abundance > 1%. In addition, several rare genera (< 1%) were grouped as others (14%). The analysis of the fungal growth morphology showed that the brine was characterized mainly by filamentous fungi life forms (55.56%), followed by yeasts (8.44%) and black yeast − like fungi (1.23%)  of ASVs was assigned to unclassified fungi. At phylum level, 56.71% of ASVs were represented by Ascomycota, followed by Basidiomycota with 8.2%, and Mucoromycota with 1%. At genus level, the most represented taxa were 25, while fungal diversity has not yet been explored extensively26. Regarding the endoglacial brines of Vida, Vanda and Bonney (west and east lobes) lakes, a number of papers have been published since the last 1990s on the biotic and abiotic characteristics e.g.29. Even in this case, however, the study of microbial communities was almost exclusively aimed at the analysis of the prokaryotic fraction.To date, researchers have mainly focused their attention on prokaryotic diversity of Antarctic brines32. However, phylogenetically the prokaryotic assemblage of BC brine showed both similarities and divergences with other Antarctic hypersaline systems. A number of phylogenetic groups (such as Cyanobacteria and Planctomycetes) that are widespread in freshwater and marine ecosystems were observed in BC brines, as well as in East Bonney and Vanda Lakes, but they were not detected in Lake Vida. Differently from East Bonney Lake, where Bacteroidetes and Actinobacteria were predominant26, Proteobacteria dominated within the bacterial communities of BC brines. The dominance of Proteobacteria (and Cyanobacteria) was also reported by Ramoneda et al.33 in Lake Vanda. However, this result referred to microbial mats, whereas water under the ice cover of the lake was characterized by a high relative abundance of Actinobacteria. In the East Bonnie Lake, Proteobacteria were the next most abundant phylum after Bacteroidetes and Actinobacteria, and mainly consisted of Betaproteobacteria, which were instead absent in BC brines.The occurrence of halophiles has been documented in worldwide hypersaline lakes: most of them belong to the phyla Proteobacteria, Cyanobacteria, Rhodothermaeota, Firmicutes, Actinobacteria, Bacteroidetes and Spirochaetes29, and in Lake Vanda34, thus highlighting their adaptation to high salinity levels. Consistently with Lake Vanda, the proteobacterial fraction of BC brines prokaryotic community included Alpha- and Gammaproteobacteria, whereas Deltaproteobacteria were not detected. The prokaryotic assemblage of BC brines also distinguished from those of Lake Vida and Blood Falls, as well as from East Bonney and Vanda Lakes, because it contains Patescibacteria as an exclusive phylotype. Patescibacteria (as well as Verrucomicrobia and Actinobacteria) are generally reported as bacterial inhabitants of permafrost (e.g.37). Interestingly, members of Patescibacteria can thrive in nutrient-limited water environments having simplified genomes which drive functions essential to growth and reproduction and retain stress response systems38. Further, the prokaryotic assemblage of BC brines (as observed for Lake Vida) harbored few Spirochaeta-related sequences unlike those of the Blood Falls brine and East Bonney Lake. These microorganisms and their metabolic features  may be useful to describe the functional ecology of briny systems.Gammaproteobacteria also occurred in the hypersaline East Bonney Lake, becoming dominant only in its deeper layers39, and Lake Vanda34. Differently from observations on Lake Vanda, sequences of Methanomicrobia detected in BC brines were most closely related to Methanoculleus (instead of the methylotrophic Methanomassiliicoccus34), a genus of methanogens commonly detected in marine and estuarine environments , but rarely reported for lacustrine habitats (e.g.41). These hydrogenotrophic methanogens well adapt to low H2 concentration, therefore possibly having an advantage over other methanogens in saline environments. Based on this result, Methanoculleus representatives are the most likely active methanogens in BC brines. Thus, differently from Lake Vanda brine, methanogenesis in BC brines occurs by the most widespread pathway, i.e. hydrogenotrophic methanogenesis, which has been suggested to be the ancestral form of methane production42.Another noteworthy feature of the BC brines concerned the occurrence, albeit scarce, of Archaea . Significant diverse archaeal populations were reported for cold briny habitats of marine origin, such as the Vestfold Hills lake system in Eastern AntarcticaMarinobacter and Psychrobacter lineages are cosmopolitan and ecologically relevant in icy brines (e.g.21). The predominance of sequences related to the Marinobacter genus makes the prokaryotic assemblage of BC brines highly similar to those reported for brines of the Lake Vida and Blood Falls21. Marinobacter members were also abundant at 30 m-depth of the hypersaline East Bonney Lake29. Marinobacter spp. are aerobic with a strictly respiratory type of metabolism. They can grow anaerobically by denitrification coupled to the oxidation of a suitable donor carbon substrate and Na+ is required for growth. Among Bacteroidetes, the genus Psychroflexus is found within moderately hypersaline ecosystems across the world, including sea-ice43, salt lake (e.g.45), salt pan46 and marine solar saltern47. Members of this genus are generally aerobic, slightly or moderately halophilic.At genus level, the gammaproteobacterial 32, the almost total absence of analyses aimed at studying the eukaryotic fraction makes difficult (or even impossible) any comparison with the other endoglacial brines found in the Vida, Vanda and East Bonney lakes. The sole comparison that can be made is that with the study of Murray et al.21, who reported that PCR surveys to detect both eukaryal and archaeal SSU rRNA genes from genomic DNA were negative, thus assuming the absence of eukaryotic microbial populations  in endoglacial brines of Vida lake. This result underlines the strong difference between them and the BC brines, where the presence of a structured fungal community was found.Considering fungal communities, despite the presence of halophilic fungi in hypersaline systems were documented22, and in another frozen lake of Boulder Clay, where the percentage of filamentous fungi and yeast form was similar26. Moreover, the high percentage of unassigned taxa (35%) found in the BC brines suggests the presence of high undiscovered fungal diversity, reinforcing the hypothesis of a high degree of biological uniqueness.In detail, the percentage of filamentous fungi found was higher (55.56%) than other growth morphology (i.e. yeasts and black yeast-like fungi), differently from the brine sampled in a frozen lake of Tarn Flat where yeasts dominated the fungal diversity49, the most abundant filamentous fungal genera colonizing the BC brines , as well as some minor genera , have been currently found in Antarctic seawaters and marine-associated habitats (e.g.51). Likewise, Glaciozyma, which is the unique yeast genus exhibiting an abundance ≥ 1%, has been widely recovered in seawaters and marine-associated habitats51. It was also found in Antarctic brines from Tarn Flat22. On the basis of these results, and in analogy to what is reported above for bacterial taxa, the hypothesis of possible marine origin of the BC brines cannot be excluded.Although some of them can be found in extreme habitats worldwide53). Their lower abundance in BC brines compared to filamentous fungi and yeasts is compatible with their lower growth rates, as reported by Canini et al.54. However, the presence of some melanized fungi  have been found in hypersaline environments worldwide32.Black yeast-like fungi exhibit morphological and physiological characteristics making them the organisms well adapted to the harsh Antarctic conditions. This microbial group is characterized by wide environmental plasticity and their ability to shift from one growth form to another, according to the physicochemical environmental conditions, may be regarded as an adaptive strategy to stressful conditions  have been pervasive on Mars, at least periodically, throughout the last 3.5 billion years, and may be present still todayConversely hypersaline waters are not so common on Earth. However, in Antarctica that is considered the best Mars analogue for many reasons, these hypersaline waters are likely more widespread, at least in the Dry Valleys.60. Therefore, their chemistry is only scarcely influenced by rock-weathering and/or biogeochemical cycles contributions61 and the possible input deriving from the dissolution of halites is poorly convincing in East Antarctica62. The cryoconcentration of seawater provides the preferential precipitation of the less soluble sodium sulfate decahydrates (mirabilite) at temperature lower than − 8.2 °C. This mechanism is considered responsible for the depletion of Na+ and SO42−. However, as observed by Cragin et al.59, the Na/SO4 ratio in brines is lower than in seawater, mainly due to the major relative changes in SO42− with respect to Na+, while Na2SO4 precipitates. In accordance with Cragin et al.59 observations, BC brines show a Na/SO4 ratio (meq/meq) of 0.022 that is about 5-times lower than seawater (i.e. 0.12). The effect of the cryoconcentration is also evident from the Cl/SO4 values reported in Fig. 4 found in BC and ELB are clearly indicative of an enhanced cryoconcentration process. In addition, Mg/K ratios, here proposed as indicators of input from non-marine sources, such as weathering, rock-water interactions as well as biogeochemical cycles, shows BC values very close to seawater composition, while ELB provides values significantly higher. Therefore, the hypersalinity of BC brines is mainly originated by seawater after an intensive cryoconcentration that likely began to occur already in remote epochs. The other peculiarity of the BC brines is the high N content that together with the total P, resulted significantly higher than other brines. This occurrence suggests a prolonged or more intense biological activity in BC brines with respect to the other ones considered in this study. Considering the advanced seawater cryoconcentration and the very scarce contribution of rock weathering in conjunction with the probable more prolonged and/or more intense biological activity, the question about where and when these brines were generated arises. At this stage, we cannot provide a comprehensive answer, but we can attempt to formulate the following innovative hypothesis. Indeed, Lyons et al.9 have already explained that the end-member brine found below the Taylor glacier , was derived from seawater and they speculated on the mechanism controlling cryoconcentration that led to the formation of the hypersaline water.The hypersalinity of Antarctic brines has been explained through different mechanisms, although there is consensus that coastal Antarctic brines mainly come from the cryoconcentration of seawater63, the glaciers of this sector of Victoria Land have preserved typical polar geomorphological features with negligible erosional power, during the last 8.2–7.5 millions of years. Apparently only the higher peaks of the Northern Foothills as the close Mt. Abbott  while, since that time, areas below 720 m asl have been repeatedly exposed and overridden by expanding ice bodies. On the other hand, if we consider Levy et al.65 at the end of the mid Pliocene, after the peak of the Pliocene warmth, when global average temperatures increased to 4 °C warmer than pre-industrial levels, a phase of tectonic uplift occurred. Consequently, the areas containing seawater and marine sediments (lagoon conditions?), where brines could have been formed, rose up. Although it is not possible to exclude that the origin of the brine can predate the Miocene, where, in some depressions, brines might had been preserved by the glacial erosion before the 8.2 Ma, even during the Early and Late Oligocene, when the Victoria Land coast was characterized by the presence of vegetation, such as Nothofagus spp., podocarps and bryophytes66.Differently, in our case the actual landscape does not show any evidence of ancient lakes or fjords or inland sea where seawater could be present in the remote past. In addition, we have to consider that according to Armienti and Baroniott Fig.  were degBoulder Clay Glacier is located BC, Fig.  just a f3−, SO42− and Cl−) and cations  were analyzed using ion chromatography (Metrohm 761 Compact IC Chromatography) equipped with a Cation 1–2  and a Metrosep Anion supp/4  column for cation and anion analysis, respectively. The brines were appropriately diluted in ultra-pure water to fit the calibration range and analyzed by ICP-MS using an iCAP RQ (Thermo Scientific) instrument equipped with an ASX-560 autosampler (Teledyne Cetac Technologies), PFA cyclonic spray chamber at 2.7 °C, sapphire injector, quartz torch, Ni cones and 1550 W of plasma radio frequency power. The ICP analysis was performed for Na, K, Ca, Mg and P. The Na, K, Mg and Ca values obtained with ICP-MS were not significantly different from those obtained with cationic IC and therefore we reported a mean value of the results obtained with these two techniques for these elements.The chemical analyses of BC brines were carried out in the laboratory of Venice. All liquid brine samples were filtered using a PTFE membrane (pore size 0.45 µm) before analyses. The anions (NO4 and Cl) were preliminarily scaled.Total carbon and nitrogen were determined with a Flash 2000 HT Elemental Analyzer (Thermo Scientific). The principal component analysis was carried out with R Studio, where the variables IlluAdp_16S_341f 5′-CCTACGGGNGGCWGCAG-3′ and IlluAdp_16S_805r 5′-GGACTACHVGGGTATCTAATCC-3′ for bacterial 16S rRNA gene region V3–V4.ii)67 for fungi.IlluAdp_ITS31_NeXTf 5′-CATCGATGAAGAACGCAG-3′ and IlluAdp_ITS4_NeXTr5′-TCCTCCGCTTATTGATATGC-3′For microbiological analyses, collected BC brines were pooled in a single sample and filtered (between 300 and 350 ml) in five replicates on polycarbonate filters . DNA was extracted from membranes using the Power Soil DNA extraction kit  according to the manufacturer’s instructions. DNA concentrations and purity were quantified by using a NanoDrop ND-1000 UV–Vis spectrophotometer . Bacterial 16S rDNA region (V3–V4) and fungal internal transcribed spacer region 2 (ITS2) were amplified using the following primers:Sequencing was performed using the Illumina MiSeq platforms, following the standard protocols of the company IGA Technology Services Srl .68. Sequences were pre-processed, quality filtered, trimmed, de-noised, merged, modeled, and analyzed via DADA2 within QIIME269 and chimeras were removed following the ‘consensus’ method reported by Callahan et al.70.FastQC was used to check the quality of raw sequences71. All sequences have been submitted to the National Center for Biotechnology Information (NCBI) under the BioProject PRJNA826749, with the biosample accession numbers SAMN27582456, SAMN27582457, SAMN27582458, SAMN27582459 and SAMN27582460 for bacteria, and SAMN27584063, SAMN27584064, SAMN27584065, SAMN27584066 and SAMN27584067 for fungi.Bacterial taxonomy annotation was performed using Silva 138 99% ASVs full-length sequences (silva-138-99-nb-classifier.qza). Fungal taxonomy annotation was performed using a Naïve–Bayes classifier trained on the UNITE + INSD database against the representative sequencesSupplementary Information.


Background: Anti-tumor immunotherapy with immune checkpoint inhibitors induces several immune-related adverse events. Camrelizumab-related isolate abducens neuritis is rare. Case presentation: We report on a 67-year-old man with esophageal cancer who presented with acute-onset isolated right abducens cranial nerve palsy after ten cycles of Camrelizumab treatment. Magnetic resonance imaging examination revealed thickening and post-contrast enhancement at the cisternal segment of the right abducens nerve. The diagnosis was immune-related abducens neuritis caused by Camrelizumab. We put him on oral taper corticoids (methylprednisone) for neuritis treatment without Camrelizumab suspension. One month after treatment, he recovered completely. At the last follow-up, one year after the onset of diplopia, the patient was in good condition without neurological symptom recurrence. Conclusion: Abducens neuritis is a rare immune-related adverse outcome of Camrelizumab. The present case proves the efficacy and safety of using corticoids in the treatment of abducens neuritis. Abducens nerve palsy, caused by various conditions, is a common neuro-ophthalmology outcome. Most cases are approximately 70 years old, and microvascular ischemia is the leading etiology . NeuritiImmune checkpoint inhibitors (ICIs) are novel immunologic agents that have revolutionized oncological practice . ICIs alIn this study, we describe a patient with esophageal cancer who manifested isolated abducens neuritis during anti-tumor immunotherapy. Magnetic resonance imaging (MRI) can localize and characterize the lesions and is an excellent modality diagnostic tool for the abducens nerve disorder. Abducens neuritis may be an irAE of Camrelizumab.A 67-year-old man with esophageal cancer presented with acute-onset horizontal diplopia for two days after ten cycles of Camrelizumab treatment. The patient had no complications of ocular pain or headaches. His ocular examination was normal. A neurological examination showed isolated right abducens cranial nerve palsy. The right eye could not abduct completely, which was confirmed by the Hess screen . An MRI The patient was diagnosed with middle-lower section esophageal cancer with lymph node metastasis one year ago  esophag.The patient had no history of hypertension, diabetes, heart disease, hyperlipidemia, cerebrovascular disease or autoimmune disease. He accepted radiotherapy and chemotherapy (Albumin-bound paclitaxel combined with carboplatin/Cisplatin/Nedaplatin). The chemotherapy drugs had to be changed frequently due to severe tertiary hemopenia, and the cancer was not well controlled until combined immunotherapy (Camrelizumab) was performed six months ago . After tWe put the patient on oral corticoids  for neuritis treatment after the diagnosis, and a subsequent taper was also administered. Prior to methylprednisone therapy, the patient was treated with enhanced microvascular perfusion drugs, but these failed to work. Camrelizumab was continued without interruption. After one week of treatment, the ability of the right eye to abduct partly recovered. One month later, the diplopia was completely resolved. . At the Importantly, potentially toxic drugs were adopted before ICIs in this case. However, paclitaxel combined with platinum was administered almost fourteen cycles before the onset of abducens neuritis, which appeared abruptly and was resolved with corticosteroids, suggesting that an inflammatory rather than toxic etiology complicated ICI treatment.Only a few case reports have reported abducens neuritis following PD-1 as possible complications , and theCranial nerve palsy caused by neuritis is characterized by thickening and post-contrast enhancement, shown under high-resolution MRI . The abdThe ICI-induced central and peripheral nerve system irAEs were found in approximately 1–3% of the patients . The posIt is challenging to balance antitumor-immunotherapy and irAE treatment. Fortunately, this abducens neuritis patient recovered quickly and showed a favorable cancer prognosis. The corticoid treatment lasted for a short duration and without antitumor immunotherapy suspension. Larger-scale and longer-term studies are needed to investigate and confirm the natural course of ICI-induced isolated abducens neuritis.Abducens neuritis is a rare irAE of Camrelizumab. The present case proves the efficacy and safety of using corticoids in the treatment of abducens neuritis.


Axons are considered to be particularly vulnerable components of the nervous system; impairments to a neuron’s axon leads to an effective silencing of a neuron’s ability to communicate with other cells. Nervous systems have therefore evolved plasticity mechanisms for adapting to axonal damage. These include acute mechanisms that promote the degeneration and clearance of damaged axons and, in some cases, the initiation of new axonal growth and synapse formation to rebuild lost connections. Here we review how these diverse processes are influenced by the therapeutically targetable enzyme SARM1. SARM1 catalyzes the breakdown of NAD+, which, when unmitigated, can lead to rundown of this essential metabolite and axonal degeneration. SARM1’s enzymatic activity also triggers the activation of downstream signaling pathways, which manifest numerous functions for SARM1 in development, innate immunity and responses to injury. Here we will consider the multiple intersections between SARM1 and the injury signaling pathways that coordinate cellular adaptations to nervous system damage. Axons enable neural communication over great distances in the nervous system. Tuned to the vulnerability of long axons, decades of studies  have showcased the plasticity of the nervous system through the varied responses made by individual cells adapting to nervous system damage. These responses include the ability to regrow damaged axons, to repair lost connections and/or sprout new axonal branches. Also critical for nervous system plasticity is the degeneration and clearance of the distal axon “stump” which has lost connectivity with the cell body.C. elegans  into Nicotinamide Mononucleotide (NAM) and cyclic (and non-cyclic) ADP-Ribose (cADPR)  . The runynthesis . SARM1 iynthesis , and alsynthesis . SARM1-mynthesis , and TLRynthesis , 2013. Iynthesis .+ loss, the products of SARM1’s enzymatic reaction may also carry out additional functions, either to facilitate degeneration or additional roles in cellular signaling. Cyclic ADPR, a robustly measurable product of SARM1 enzymatic activity, is a known regulator of the ER Ryanodine receptor and TRPM2 calcium channels  could be observed to coalesce into visible puncta during conditions of stress that activate its downstream signaling  was shown to be responsible for SARM1 activation, and subsequent SARM1-mediated axonal degeneration. This SARM1 activation can be rescued by inhibiting MAP Kinase/JNK signaling  are known to be activated from multiple aspects of axonal injury response  could inhibit degeneration caused by ectopic activation of SARM1’s TIR domains . The duaOne more set of twists in the intertwined relationship of JNK signaling with SARM1 come from observations in invertebrates that JNK signaling activation following axonal damage can have protective functions in neurons. This includes the inhibition of degeneration in both axons and dendrites , which cDrosophila  kinases. WNK kinases are broadly expressed, best known for roles in the regulation of ion transport and blood pressure regulation . Howeverutgrowth . KnockdoThe SARM1 NADase enzyme is an attractive druggable target for intervening neuropathies and a variety of neurodegenerative diseases. SARM1’s enzymatic activity drives the degeneration of axons and, in addition, regulates signaling pathways that influence development, inflammation, and multiple responses to axonal injury. Further consideration of SARM1 as an enzymatic target requires a deeper understanding of SARM1’s roles in development and the crosstalk of SARM1’s multiple functions in the context of nervous system injury.TW and CC worked in collaboration to carry out the conception and writing and revising of this review. Both authors contributed to the article and approved the submitted version.


Giardia duodenalis is an important zoonotic protozoon, which can infect a variety of animals, causing diarrhea and even death of animals or humans. Dairy cattle have been implicated as important sources of human G. duodenalis. However, the information about the prevalence and genetic diversity of G. duodenalis in dairy cattle in China's Yunnan Province remains limited. This study investigated the occurrence and multilocus genotyping of G. duodenalis of Holstein cattle in Yunnan Province, China. A total of 524 fresh fecal samples of Holstein cattle were randomly collected from 8 farms in Yunnan. In this study, 27.5% (144/524) of tested samples were positive for G. duodenalis infection. The highest infection ratio was found in preweaned calves (33.7%), and the infection rates of postweaned calves, growing cattle, and adult cattle were 24.5%, 23.0%, and 17.3%, respectively. The sequence analysis of SSU rRNA gene showed that the predominant assemblage of G. duodenalis in this study was assemblage E , whereas assemblage A was identified only in three samples . All G. duodenalis-positive samples were further assayed with nested polymerase chain reaction (PCR) targeting β-giardin (bg), triosephosphate isomerase (tpi), and glutamate dehydrogenase (gdh) genes, and 87, 41, and 81 sequences were obtained, respectively. Mixed infection of assemblages A and E of G. duodenalis was detected in three samples. Multilocus genotyping yielded 23 multilocus genotypes (MLGs). This is the first study that reveals the prevalence data of G. duodenalis in Holstein cattle in Yunnan Province, and the results of this study provided baseline data for the prevention and control of G. duodenalis infection in Holstein cattle in Yunnan Province, China. Giardia duodenalis is one of the most common parasitic protists that can infect humans, livestock, companionn animals, and wildlife   –4. Trophnd cyst) , 6, and 25 cysts . G. duoded hosts , and theG. duodenalis consists of eight assemblages (A–H), and some of those aggregates display host specificity   , assemblbovines) . Recentlbovines) –16. DairB, and E . Calves B, and E . G. duoddiarrhea . The inf2 to 89% –27. Therin China –36. Humaia cysts . It is cd cattle , 39.G. duodenalis. Mixed infection of G. duodenalis will result in inconsistent genotyping results of different loci for that bg, tpi, and gdh genes show high genetic polymorphism  targeting the small subunit ribosomal RNA (SSU rRNA) gene of G. duodenalis. All positive samples were further subjected to the gene analysis of bg, tpi, and gdh genes for the genotyping of G. duodenalis.Up to now, the infection data of This study was approved by the Life Science Ethics Committee of Yunnan Agricultural University with the ethical code 202109003. Fecal samples were collected from the Holstein cattle with the permission of the farm owners or managers.From July to November 2021, a total of 524 fecal samples of Holstein cattle were randomly collected from 8 farms, including one free-ranging farm and seven intensive feeding farms. The age of Holstein cattle ranged from newborn to 2 years old. The collection sites included Dali, Kunming, Qujing, and Chuxiong. Feces were collected from 422 females and 102 males, and only 18 of them had obvious clinical symptoms of diarrhea. Fresh fecal samples (10–20 g per cattle) were collected directly from the rectum using disposable gloves and then transferred separately into disposable plastic bags, marked with the date, age, and geographical information. Fecal samples were stored at 4°C until used for DNA extraction.The age of Holstein cattle was classified according to the Technical Specification for Standardized Scale Breeding and Production of Dairy Cows  issued by the Ministry of Agriculture of the People's Republic of China. The cattle ≤ 60 days old are preweaned calves, 61–180 days old are postweaned calves, 181–450 days old are growing cattle, and ≥ 450 days old are adult cows.g for 3 min; 250 mg of each washed sample was used for DNA extraction individually. The DNA of the collected sample was extracted by using E.Z.N.A.R® Stool DNA Kit  according to the manufacturer's instructions. The extracted DNA was stored at −20°C until use.Before DNA extraction, stored feces were washed with distilled water and centrifuged at 3,000 × G. duodenalis to determine the infection of G. duodenalis , and 2 μl of DNA sample. Dimethyl sulfoxide (DMSO) was added to enhance the amplification efficiency of nested PCR. The products of the second nested PCR were subjected to electrophoresis on 1% agarose gel and photographed by using a gel imaging system.All DNA samples were tested with nested PCR that targets the SSU rRNA of odenalis . The bg odenalis , tpi  Co., Ltd. for bidirectional sequencing. All sequences obtained in this study were searched against GenBank by using BLAST, and the Clustal X software was used for sequence alignment analysis. All representative nucleotide sequences generated at bg, tpi, and gdh loci in this study were deposited in the GenBank database under accession numbers ON773555–ON773581.2) test was used to analyze the differences in G. duodenalis infection in Holstein cattle among different regions, age, sex, and farming methods. The confidence interval was set as 95%, and P < 0.05 was considered statistically significant. All statistical analyses were performed by using the SPSS20.0 statistical software.The chi-square  . Infection of G. duodenalis was found in Holstein cattle in all age groups, among which the highest infection rate (33.7%) was found in preweaned calves. Adult cattle showed the lowest infection rate (17.3%). The χ2 test showed that the prevalence of G. duodenalis among the four age groups was significantly different      . G. duodto 43.7% . The hig < 0.01) . No sign > 0.05) . By comp < 0.05) . In addi > 0.05) .G. duodenalis assemblage A  and the rest of them were a member of assemblage E . Of the 144 positive fecal samples, 60.4% (n = 87) were bg positive, 28.5% (n = 41) were tpi positive, and 56.3% (n = 81) were gdh positive. Notably, 20.1% (n = 29) of samples were positive for all four genes in this study.Sequence analyses of the amplified 144 SSU rRNA genes showed that three of them were classified into n = 8), E3 , E5 , E1 , and E2 ; the remaining 4 subtypes represent novel subtypes (n = 9), E17 , E2 , and A1 . The remaining 13 subtypes of assemblage E and 1 subtype of assemblage A represented novel sequences. At the gdh locus, 13 subtypes of assemblage E and 1 subtype of assemblage A were identified, including 5 known subtypes of assemblage E with sequences identical to those in GenBank, namely, E3 , E1 , E14 , E15 , and E . The remaining 8 subtypes of assemblage E and 1 subtype of assemblage A represented novel subtypes in this study , E34 , and E3  were the predominant subtypes.The bg subtype analysis showed that 9 subtypes of assemblage E and 1 subtype of assemblage A were identified, among which 5 subtypes of assemblage E had previously been identified; for that, there was a 100% similarity to the sequences available in GenBank with accession numbers of E9 . Three samples showed mixed infection with assemblages E and A   observed in this study was higher than that of most provinces in China, such as Jiangsu    , Henan  , Xinjian1/1,366) . It was 1/1,366) , Tibetan1/1,366) , and Yun1/1,366)  but lowe1/1,366) , Guangdo1/1,366) , and Sha1/1,366) . Compare 45/900) , South K 45/900) , 53, Ira 45/900) , and Egy 45/900) . InfectiG. duodenalis is associated with animal age , which is consistent with the findings of a previous study in Sichuan  and contrary to the prevalence data of G. duodenalis in Hubei  were detected in dairy cows, and assemblage E was the dominant assemblage in this study. In other studies, assemblage E was also the predominant genotype in dairy cows  have been found in iry cows , 35, 36.iry cows , Brazil iry cows , and Egyiry cows , 65 has G. duodenalis. A total of 9 assemblage E subtypes and 1 assemblage A subtype were identified by using bg locus, 16 assemblage E subtypes and 2 assemblage A subtypes were identified by using tpi locus, and 13 assemblage E subtypes and 1 assemblage A subtype were identified by using gdh locus. The combination of sequence polymorphisms at these three loci led to the identification of 23 E MLGs, and three samples had different assemblages at three loci  of G. duodenalis were found in Holstein cattle, and assemblage E was identified as the dominating genotype. The presence of zoonotic assemblage A in Heqing and Shilin cattle suggests their zoonotic potential. Multilocus genotyping at bg, tpi, and gdh loci revealed 23 novel assemblage E MLGs and 3 E+A mixed infection in Holstein cattle. These findings indicate that G. duodenalis of Holstein cattle in Yunnan is rich in genetic diversity, and the sequence of each gene locus is quite different. For the limited sensitivity of PCR, intensive study is required to reveal the molecular characteristics of G. duodenalis in Holstein cattle in Yunnan, and it is important to strengthen the surveillance of this parasitic disease to ensure the health of livestock and human beings.This is the first MLG characterization study of The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found in the article/supplementary materials.The animal study was reviewed and approved by the Life Science Ethics Committee of Yunnan Agricultural University.H-MM, J-FY, and J-JH designed the study. J-FY performed fecal sample collection. Z-JH, X-YX, C-RX, and J-RC performed the molecular genetic studies. JM analyzed sequences. J-JH, J-FY, and H-MM provided laboratory supplies and revised the manuscript. Z-JH interpreted the results and wrote the manuscript. All authors have read and approved the final manuscript.This study was funded by the Scientific and Technological Mission of Dairy Cow Industry in Heqing County, Yunnan Province (Grant No. 202204BI090005), the Veterinary Public Health Innovation Team of Yunnan Province (Grant No. 202105AE160014), and the Yunnan Expert Workstation (Grant No. 202005AF150041).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Patch clamp technique was used for measuring transient receptor potential melastatin 2 (TRPM2) channel. Ligation of left anterior descending for 2 h was performed to induce MI animal model. The rats were treated by different concentration of uric acid. The artery tissues were stained by HE and collected for measurement of NLRP3 and inflammatory factors. Supplementation of UA significantly promoted apoptosis, and augmented the expression of intercellular adhesion molecule-1, chemoattractant protein-1, vascular cell adhesion molecule-1, and NLRP3 inflammasome. Knockdown of NLRP3 reversed the influence of UA on MI by decreasing collagen deposition, fibrotic area, apoptosis. The expression of NLRP3 inflammasome increased markedly after treatment of UA. UA activated ROS/TRPM2/Ca2+ pathway through targeting NLRP3. UA activated NLRP3 inflammasome and augments inflammatory factor production, which in turn exacerbates cardiomyocytes injury. Knockdown of NLRP3 reversed the influence of UA on apoptosis and cell cycle. UA may promote cardiomyocytes injury through activating NLRP3 inflammasome and ROS/TRPM2 channel/Ca2+ pathway.Cardiomyocytes injury has been considered as a key contributor for myocardial infarction (MI). Uric acid (UA) can induce cardiomyocytes injury, which is closely related to NLRP3 activation and inflammatory factor generation. However, the mechanism how UA modulates cardiomyocytes remains elusive. Western blotting and qRT-PCR were applied for measuring protein and mRNA expression, respectively. ROS production and CaThe online version contains supplementary material available at 10.1186/s12872-023-03040-1. Cardiovascular disease is a major public health problem affecting human health. Myocardial infarction (MI) is one of the main causes of sudden death. Although the method of rapid recovery of coronary blood flow is commonly used in clinic, which can restore the blood supply to the myocardium and improve the survival rate of patients with myocardial infarction. However, this method is not enough to prevent the further deterioration of cardiac function and might further cause serious problems such as ischemia–reperfusion injury. It has been reported that high uric acid (UA) is closely related with cardiovascular disease, and the severity of cardiac lesions is positively related to the level of serum UA , 2. MeanPyrin domain-containing 3 (NLRP3) inflammasome is a multiprotein complex comprising NLRP3 receptor protein, protease caspase-1 and apoptosisassociated specklike protein (ASC) , 8. NLRP2+ influx, and also known as a sensor for oxidative stress and ROS [2+ influx through TRPM2 channel has been considered as an important regulator for inflammatory factors [2+ remains unclear.Oxidative stress has been believed as a key contributor for the atherosclerosis, and the reactive oxygen species (ROS) induced by oxidized low-density lipoprotein could initiate endothelial dysfunction and further accelerate atherosclerosis . transie and ROS . ROS ind factors . However2+.This study may provide a new insight about the intervention and prevention of cardiomyocytes injury and MI.In the present study, we investigated the influence of UA on the cardiomyocytes apoptosis. The effects of UA on the expression of MCP-1, VCAM-1, ICAM-1, and NLRP3 inflammasome were measured. Additionally, we conducted animal experiments to observe the morphological changes of myocardial tissue after treatment with different concentrations of UA. Finally, we proved that UA promote cardiomyocytes injury via upregulating NLRP3 inflammasome and activating ROS/TRPM2/Ca2. After reaching 80% confluence, the cells were passaged with 0.3% trypsin–EDTA and seeded onto plats. After different treatment describe below, the cells were applied for different experiments. The cardiomyocytes used in this study were identified with cardiac markers (connexin 43 and SIRP-a), and presented in Additional file Human cardiomyocytes  and human aortic endothelial cells  were incubated with DMEM  containing 10% FBS , 90 g/mL heparin, and 1% penicillin/streptomycin at 37℃ and with CO6 cells were suspended in 200 µL binding buffer containing 5 µL propidium iodide and 10 µL Annexin V-FITC. Incubate cells in the dark for 30 min, and flow cytometry analysis  was conducted.These experiments were performed as described previously , 19. FirThe cell cycle was performed as described below. Cells were treated with UA or siRNA for 24 h. The cells were digested with trypsin (0.2%). After fixation with 70% cold ethanol, the cells were incubated with RNase A for 20 min. Then, the cells were incubated with propidium iodide for 30 min in the dark. Cell cycle was analyzed with flow cytometry . Flowjo™ software v11 was used for flow cytometry data analysis. The gating strategy was set as follows. The cells without staining by PI and Annexin V-FITC were used for flow cytometry analysis. The gate was set to cover negative cells in the lower left quadrant. Then, the set gate was used to analyze apoptotic cells and dead cells in the lower right quadrant and higher right quadrant, respectively. FSC-H versus FSC-A strategy was used to discriminate doublets. The experiments were repeated at least 3 independent times. 3 samples were set in each group.The cell proliferation was measured as described previously . The proThe western blotting was performed as described previously . The priThe experiment was conducted following the method of previous study . BrieflyThe supernatants were collected and centrifuged to remove cell debris. MCP-1 , VCAM-1 , and ICAM-1  in the supernatant were detected with relative ELISA kits. The experiments were repeated at least 3 independent times. 3 samples were set in each group.All the experimental protocols were approved by Independent Ethical Committee of Quanzhou First Hospital Affiliated to Fujian Medical University. Wistar rats  were anaesthetized by intraperitoneal injection with ketamine (120 mg/kg) as describe previously . The aveAnimals were euthanized at 4 weeks after surgery with prolonged exposure to isoflurane as described previously . Tissues2+ in cell was measured in the similar way as described above except that cells were labeled with 10 μM Fluo-3AM  for 1 h. After washing with buffer free of calcium ions, cells were measured by flow cytometer  at an excitation wavelength of 480 nm. The experiments were repeated at least 3 independent times. 3 samples were set in each group.Cells were labeled in the dark with 2 mM DCFHDA–AM  for 20 min firstly. After washing and re-suspending in PBS , the cells were analyzed by flow cytometer  at an excitation wavelength of 514 nm. The CaThe TRPM2 channel current measurement was performed as described previously . EPC-10 Transthoracic echocardiography examination was performed uing a Xario ultrasound device  with a 12 MHz cardiac probe after animal anesthesia. MP100-CE  was applied for recording. The experiments were repeated at least 3 independent times. The model establishment was confirmed by the occurrence of ST-segment elevation in electrocardiogram. The ST-segment elevation electrocardiogram was presented in the Additional file The sections were incubated with proteinase K , hydrogen peroxide , and Triton X-100 , and washed with PBS (5 min). 5% goat serum  was used for blocking, and BCIP/NBT liquid  was used for incubating Sects. (2 h). The sections were mounted and observed with a fluorescence microscope . At least 5 random images in each group were used for apoptosis analysis.P values < 0.05 was considered statistically significant.Assays were performed at least in three independent experiments with one representative picture presented. Data were presented as mean ± standard deviation (SD). Statistical analysis was performed using Graphpad Prism 4.0 . One-way analysis of variance (ANOVA) was applied for Statistical evaluation among the different groups. As shown in Fig. In view of the crucial role of NLRP3 inflammasome in modulation of inflammation, we assessed the status of NLRP3, Cacspase-1, and ASC followed treatment by different incubation time or different concentrations of UA Fig. . We founTo determine whether UA treatment enhanced pro-inflammatory cytokine production, we analyzed various cytokines secretion from cells treated by UA Fig. E, F for Knockdown of NLRP3 vector was constructed with siRNA-NLRP3 (si-NLRP3), and the knockdown of NLRP3 was validated in vivo and in vitro levels Fig. A, B. How2+ pathway through targeting NLRP3.It was reported that, high ROS production , TRPM2 cThe effects of si-NLRP3 on cell apoptosis and cell cycle in vitro were also investigated with both AC16 and HAEC cells. UA treatment significantly increased apoptosis rate and cell percentage in the S stage, but decreased cell percentage in the G2 stage compared to group control  and not in rat adult cardiomyocytes, which is a major limitation. In addition, the apoptosis and necrosis seen in MIRI might be caused by ischemic injury resulting from thrombosis of the atherosclerotic coronary vessel, which needs to be further studied.Previous study indicated that TRPM2 channel/Caammation . Meanwhidiseases , and thediseases . We firsChronic kidney disease is a known atherosclerotic risk factor, and also leads to hyperuricemia. It was reported that UA levels were significantly higher in patients with acute myocardial infarction, and hyperuricemia was closely associated with acute myocardial infarction . In addi2+ pathway. These findings provide specific evidences about how UA influences the cardiomyocytes injury. Therefore, the control of serum UA should be emphasized and intervened in patients with MI risk. In the primary care setting, patients with MI risk should be tested for hyperuricemia and treated to prevent worse outcomes. This research provides a novel thought for the therapeutic strategy of MI injury by regulating NLRP3/ROS/TRPM2/Ca2+.The findings of the present study show that UA treatment activates NLRP3 inflammasome, promotes cell death, and increases the production of inflammatory factors, thereby leading to MI. Additionally, UA may contribute to cardiomyocytes injury through activating NLRP3 inflammasome and ROS/TRPM2/CaAdditional file 1. Supplementary Figure 1. Identification of cardiomyocytes with connexin 43 and SIRP-α antibodies.Additional file 2. Supplementary Figure 2. Full-length blots/gels of Figure 2 A.Additional file 3. Supplementary Figure 3. Full-length blots/gels of Figure 2 C.Additional file 4. Supplementary Figure 4. Full-length blots/gels of Figure 3 A.Additional file 5. Supplementary Figure 5. ST elevation electrocardiogram data.


It has been a challenge to support the expansion of urban agriculture (UA) in cities due to its poor economic profitability. However, it is also hard to deny the increasing benefits of UA in improving the socio-environmental dimension of cities. Hence, in this review, different aspects of UA were examined to highlight its value beyond profitability such as social, health and well-being, disaster risk reduction, and environmental perspectives. A case study and relevant policies were analyzed to determine how policy makers can bridge the gap between current and future UA practices and sustainable development. Bridging these policy gaps can help the UA sector to sustainably grow and become successfully integrated in cities. Moreover, advancements in UA technologies and plant biotechnology were presented as potential solutions in increasing the future profitability of commercial UA. Consequently, as new UA-related technologies evolve, the multidisciplinary nature of UA and its changing identity from agriculture to digital technology, similarly require adaptive policies. These policies should maximize the potential of UA in contributing to resiliency and sustainability and incentivize the organic integration of UA in cities, while equally serving social justice. Urban agriculture; Vertical farming; Sustainable city; Policy; Genetically modified plants. It has the potential to end world hunger and boost the economies of developing nations. It is also an essential industry that will remain at the center of human activity for many centuries to come. However, the agricultural system practiced today can hardly be called sustainable due to the increasing strain it puts on our planet's scarce resources. Especially with the growing population projected to peak at nearly 11 billion by 2100, agriculture will struggle to meet the needs of the world population . To shouPreliminary to addressing the contemporary interests in UA, it should be noted that though not as readily exemplified in the modern, developed world, agriculture does in fact have a longstanding history in urban spaces . Take foWhen placed in modern discourses however, UA has evolved as an effective tool and commonly cited solution to many contemporary challenges. The Sustainable Development Goals (SDGs) set out by the United Nations for the year 2030 is a direct manifestation of the types of initiatives in which UA can be employed for developed and developing countries alike. The growing number of urban farm initiatives may be attributed largely to its importance in food security efforts. This has coincided with a simultaneous emergence of local food production movements in developed countries with populated metropolitans . That isAlongside food security, and as this review will also seek to explore, urban and community farming efforts encompass a wider range of beneficial services which require appropriate implementation. In many cities, community farms have offered alternative social benefits to the residents. For example, a study conducted on farms in New York found a wide range of shared goals exhibited by the local farmers. Significantly, it outlined the numerous ways in which practitioners contributed to social, political, economic, and environmental problems external to food production. Some such activities included educational programs and workshops on health and nutrition, environmental restoration, and political activism within the realm of UA .On the contrary, when coupled with the UA's recent resurgence, the ramification of such diverse approaches and experiences has been the UA's incompatibility with a rather narrowly defined legislative system. This has in turn slowed down or completely inhibited the incorporation of initiatives into cities . On the Even in instances where UA is not actively contributing to gentrification processes, farms have run into other problems. For instance, small communities or family farms often utilize labor-intensive methods because of a lack of access to necessary equipment or limited awareness of more efficient alternatives. Economic viability is thus compromised due to the low efficiency of material and labor inputs . ConversThis paper has conducted an integrative review of the literature to identify the multifarious aspects of UA and how these have directly or indirectly contributed to the viability of its application. It seeks to update and contribute to the UA topic by employing a multi-perspective approach and providing an integrated look into UA as a whole . To achiThe paper adopts a holistic approach by considering both theoretical and empirical research, with each perspective offering alternative insights into the potentials and perils of UA implementation. It therefore aims to provide an overview and analysis of relevant literature that is available to date. To this extent, the recommendations are based on and limited to the conclusions drawn by selected literature. Further empirical research would thus be required to substantiate these claims and better assess the practicality of implementation.2experimenting with radical alternatives to the capitalist neoliberal organization of urban life”  was measured in a bubble-insulated greenhouse, compared to typical conventional greenhouses which have a heat loss of about W/m2 K. This makes the energy demand for the small-scale greenhouse lower of only 10–20 % of the energy consumed  by conventional Nordic greenhouses. Also, the incorporation of anaerobic digestion is advantageous in upcycling the organic agricultural wastes such as the roots and stems of crops, which are regularly produced after each harvest in the farm. However, further studies are needed to successfully up-scale the system and optimize growing conditions of crops in terms of substrate microbiology.One remarkable application is the ‘Food to waste to food’ project which was claimed to be the first efficient method for the utilization of digestate as a growing medium and bio-fertilizer in greenhouses . This prunpublished). Upon commercialization, anaerobic digestion system can easily be integrated to UA, providing better efficiency to any types of farming system. The same compact system platform can also provide wastewater treatment function to remove excess fertilizer before a necessary water disposal.On the other hand, seamless and compact biogas digester design which can be operated in urban setting while not compromising energy production is already under development , genetic modification (GM) or genome editing (GE). In MAS, conventional breeding can be made faster by using DNA markers to select for hybrids instead of using phenotypic selection which usually requires longer periods of time. On the other hand, GM involves the use of recombinant DNA technology to change the genetic makeup of organisms. Recombination is the insertion of DNA molecules from different distinct species to produce an improved version of the organism. Finally, GE is the most recent technology that has shown immense potential for application in plant biotechnology. Genome editing is based on the precise identification of short DNA sequences and their deletion, then insertion of new DNA sequence to correct errors or to change the genetic information.CRISPR-Cas9, Using plant biotechnology tools, it is now possible to develop crops with desired traits such as resistance to pests and diseases, tolerance to drought, heat, cold or salinity, improved flavor, rapid cycling as well as other superior growth traits. For urban agriculture, the limited space for cultivating crops can be addressed by developing plants with compact architecture and rapid life cycle . Using tWhile crops that are developed by conventional breeding are more easily accepted by the public, those that involve GMs are still not accepted by the Japanese consumers even though the Japanese government has approved the commercial cultivation of GM crops. The same is true for genome edited agricultural products. However, it is interesting to note that Japan remains one of the top importers of food and feed products developed using genetic engineering from the US. According to USDA Foreign Agricultural Service report, Japan imports 100 % of its corn supply and 94 % of soybean supply, which are mostly GM . Genome 2.3“emphasise a balance among economic development, environmental protection, and equity in income, employment, shelter, basic services, social infrastructure and transportation” became prominent  household level, (b) city level, and (c) macro level .At the household level, economic benefits and costs involved in agricultural production such as self-employment, exchange of products, income from sales, savings on food and health expenditures are directly incurred by the urban households. A study in four West African capitals showed that rainfed crops such as maize and cassava are mainly produced for household consumption, while short-cycle and long-cycle crops such as lettuce, cabbage, carrots, and onions can generate monthly income from sales . FurtherAt the city level, there are: (a) direct benefits and costs which are not carried by the farmers, and (b) indirect benefits and costs which are in the form of positive and negative externalities. These externalities include the social, health, and environmental impacts of UA in the urban setting. However, comparing different city situations remains a challenge as these impacts depend on policies and legislation existing in the city. One common approach for economists to examine or quantify these impacts is by using cost-benefit framework  althoughAt the macro level, effects of UA are felt through its contribution to the national's gross domestic product (GDP) and to the efficiency of the national food system. Moreover, UA products can supplement rural agriculture's limited supply, substitute for food imports, and boost export production of agricultural commodities . In KenyThe term ‘economic viability of UA’ can also be ambiguous. Copious literature has discussed the economic viability of either micro-farms, rooftop gardens, greenhouses, or vertical farms to examine the cost and gains of these specific types of UA . These a3.2Regarding the implementation of UA systems within developed countries it is important to acknowledge that integration is taking place within pre-established socioeconomic structures, and not the other way around. In the Global North, the physical and cultural environments encountered by the UA narrative are often distinguished in part by deeply rooted societal structures and potential injustices requiring attention. To this end, systems of inequality can distort the “sustainable” and “social justice” front commonly adopted by UA initiatives, by engulfing operations within socially detrimental processes like eco gentrification . In othea corporate food system model of profit maximization and resource use efficiency, subscribing to capitalist logics rather than alternative, social-justice-oriented practices”  disregard for precautionary limitations, 2) misuse of authorized active substances, and c) use of unauthorized substances.” While these are preventable, the lack of supervision and regulation on backyard UA may suggest that the risks are still present albeit minimal. To avoid risks, it is highly recommended to regulate the type and size of farming in cities. For example, mandating the use of vertical farm technique when establishing a commercial-scale UA will prevent these PPP risks in urban communities while bringing commercial-scale source of food within cities.Environmental risks often emerge as agricultural practices shift into city centers. Such risks may pertain to the production of goods and services by farms, or they may appear as negative externalities in the surrounding community. For instance, increased levels of pollution in cities can diminish the quality of urban-grown products, generating health risks for consumers . MeanwhiThe relationship between the agricultural sector and the environment is defined in two senses by the latter. Namely, the environmental impact induced through alterations made by farming practices, and subsequently the kind of environment that is produced by incorporating food production in the given region. This is true of urban and rural systems alike, illuminating the push to reconcile modern agrarian methodologies with environmental conscious regulations . RegardiOne of the most prominent environmental risks faced by UA in contemporary societies has been the navigation and risk management associated with environmental contamination. Specifically, the anthropogenic pollution of soil and air as a result of industrial activities, transportation, mining, sewage, and fossil fuel combustion. The ultimate impact of environmental contamination of produce is dependent on several factors such as the quantity and type of pollutant present, how long the produce remains in the soil, and similarly the kind of crop being exposed. Vegetables like lettuce and cabbage risk greater exposure to atmospheric particles on account of the greater surface area of leaves, while root vegetables are more vulnerable to soil contaminants. Duration of growth will also increase or reduce the amount of exposure to any pollution present, and so herbs like thyme, which are grown year-round, become more susceptible to absorption .Regarding contamination, lead is a commonly cited concern for urban farmers utilizing soil-based methods of crop cultivation. Leaded-gasoline and paints were widespread several decades ago, despite the phase-out of such products many urban sites today continue to test positive for varying levels of contamination . HoweverStill other forms of urban air and soil pollution do exist that could impede more seriously upon the uptake of UA systems in certain cities. For instance, old industrial sites may be more prone to different forms of contamination depending on the type of activities once conducted on the land . While pHowever, these risks may be subverted depending on the type of UA that is being utilized. For instance, farming technologies associated with indoor farming, hydroponics, aquaponics, may help to minimize the impacts of soil and/or air pollutants generated by human activity. Though the use of alternative farming mechanisms can help mitigate risks posed by urban pollution, its employment is succeeded by other changes in production that can affect the overall economic viability and sustainability of UA. A simple example of this might be how the use of indoor farming shields crops from air and soil pollution in cities, but may simultaneously require greater energy consumption for climate control systems .From the opposite perspective, agricultural practices which are focused in producing high-quality products, especially those which are utilizing terroir approach, will be more inclined to improve the environment and local ecosystem condition where the UA are located. Using terroir concept, the interaction between local environment and ecosystem characteristics as well as the local agriculture knowledge and practices can directly influence the characteristics of agricultural products . In the 4To accommodate the multiple functions of UA, in addition to the sector's intrinsic diversity, respective urban policies require a degree of structural robustness in ensuring proper integration. By and large however, policies remain limited in scope, and incapable of sufficiently implementing systems within respective municipalities . The morwithout explicit valuation of food justice” policy mechanisms existing congruent to this common, well-researched stance will ultimately fall short of uplifting the disadvantaged communities they seek to target. Additionally, UA is only part of a food justice solution, and that “there is a distinction between alleviating symptoms of injustice . . . and disrupting social and political structures that underlie them”  the farm utilizes various advanced technologies to help ensure high output efficiency alongside year-round production. This is inclusive of computer systems that manage internal temperatures and irrigation. Moreover, the installation of solar photovoltaics, advanced ventilation systems, and high efficiency pumps and fans further seeks to optimize energy efficiency of the greenhouse . Since iConstruction of the flagship farm at Greenpoint was completed in 2011 following the introduction of new zoning regulations within the state of New York. Specifically, those that enabled Gotham Greens to secure zoning approval eliminated height and bulk restrictions that had previously affected rooftop farms and gardens in the city. Changes in said laws emerged in 2010 in response to increasing awareness for UA initiatives, and particularly sought to encourage and accommodate the development of CEA in urban areas .Policy development in favor of vertical farms is reflective of a trend in the recent decade to support farms associated with high-tech systems like that of Gotham Greens. Accordingly, this resulted in the emergence of other CEA farms around the same time in New York, including Brooklyn Grange, Eaglestreet Rooftop Farm, and Square Roots to name a few . The driIn terms of its economic viability, Gotham Greens has been recorded to “produce 7–8 times more food than traditional farming” on account of its technology-dependent efficiencies, and year-round production. Coupled with the fact that the organization was the only supplier of fresh food during Hurricane Sandy, these characteristics appear promising in the context of food security . HoweverRegarding employment, policies exist for the promotion of green spaces in the city for ecological-environmental, aesthetic-recreational, and social-educational purposes.” One study conducted in the United States found that between the years 2001 and 2007, a total of 204 bills related to park improvement and green space support were passed. The bills covered a wide range of dimensions including, funding, outreach, preservation, recreational activities, and safety. The diversity and quantity of bills passed were thus indicative of “a continued commitment to improvement and reinvention of existing policies” in the states represented by the study  which assists and supports the state's smaller scale urban food producers . AdditioThe achievement of high sustainability in urban farms is contingent upon the training and knowledge procured by producers. This contrasts the tendency of recreational farmers to make less sustainable choices, resulting in low efficiency of material and labor inputs . This maConversely, while local governments can bolster productivity and sustainability of UA, education becomes another benefit of integration as awareness is generated amongst residents concerning topics like nutrition and food production . Promoti5The economic profitability of UA is highly dependent on its size, type, price competitiveness, and consumers’ perceived value of produce beyond uses as food. Despite its highly relative profitability, UA has many different roles for communities in cities and urban areas, from subsistence-oriented motives to large scale commercial production facilities. Through UA, a household can reduce its expenses by producing its own food, thus leading to savings in their household budgets . FurtherOn the other hand, UA has a potential to be economically profitable as a commercial-scale food producer in a closed system and controlled environment such as vertical farms, plant factories, and greenhouses . The tecWhile the resurgence of UA among cities worldwide has been mainly driven by the public and private sectors, the role of policy makers is an integral part of UA revolution to successfully integrate UA practices in cities. Existing policies and regulations, land prices, availability of urban markets, as well as the prices for agriculture commodities strongly influenced UA activities . Its curAll authors listed have significantly contributed to the development and the writing of this article.This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.Data included in article/supp. material/referenced in article.The authors declare no conflict of interest.No additional information is available for this paper.


Phytohormones are major signaling components that contribute to nearly all aspects of plant life. They constitute an interconnected communication network to fine-tune growth and development in response to the ever-changing environment. To this end, they have to coordinate with other signaling components, such as reactive oxygen species and calcium signals. On the one hand, the two endosymbiotic organelles, plastids and mitochondria, control various aspects of phytohormone signaling and harbor important steps of hormone precursor biosynthesis. On the other hand, phytohormones have feedback actions on organellar functions. In addition, organelles and phytohormones often act in parallel in a coordinated matter to regulate cellular functions. Therefore, linking organelle functions with increasing knowledge of phytohormone biosynthesis, perception, and signaling will reveal new aspects of plant stress tolerance. In this review, we highlight recent work on organelle–phytohormone interactions focusing on the major stress-related hormones abscisic acid, jasmonates, salicylic acid, and ethylene. A review of organelle–phytohormone interactions and retrograde signals, highlighting recent work and open questions and focusing on the stress-related hormones abscisic acid, jasmonates, salicylic acid, and ethylene. Organelles have essential functions in most cellular processes including growth and development of plants. As a consequence of their endosymbiotic origin, mitochondria and chloroplasts carry their own genomes. However, the majority of organellar proteins are now nucleus encoded . Work haIn addition to their role as environmental sensors, organelles  are the main hub for the metabolism of several phytohormone precursors, as illustrated in Classically, hormone signaling is discussed in a historical context where specific hormones are often viewed in a particular developmental or stress context. For this review, however, it seemed more reasonable for us to structure organelle–hormone signals to cover (i) direct signaling connections, (ii) indirect signals, and (iii) other, more complex signaling interactions . Direct In this review, we summarize recent work on organelle–phytohormone interactions to highlight and address open questions within these interwoven signaling cascades, concentrating on the stress-related hormones ABA, jasmonates, SA, and ethylene. However, other phytohormones also have tight relations to plant organelles, and the impact of auxin, cytokinins, gibberellins, and strigolactones on photosynthesis and photoprotection was covered in a recent review by ABA was identified in the 1960s as an endogenous signal (cis-epoxycarotenoid dioxygenase (NCED), which leads to xanthoxin. The oxidative cleavage via NCED is the first irreversible conversion step towards ABA and well accepted as a bottleneck in ABA biosynthesis -1,1-dimethylurea, paraquat, and ascorbate modifies the pace of nuclear-driven circadian oscillations , an important component of the molecular clock in plants , encoding a highly abundant reactive oxygen species (ROS) scavenger in chloroplasts, revealed that its expression is controlled by ABA together with the cellular redox state . The majority of photosynthesis-related nucleus-encoded genes are repressed by ABA treatments (gun1-1 and abi4 knockout plants and suppression of the gun1-1 phenotype in ABA INSENSITIVE 4 (ABI4) overexpression plants, it was proposed that ABA signaling via the transcription factor ABI4 is an important downstream component of GUN1-mediated chloroplast biogenesis  questions a role of ABI4 in GUN1-mediated biogenic control of photosynthesis-related, nucleus-encoded genes (ABI5 overexpression suppresses chloroplast development and expression of photosynthesis-related genes in potato (Solanum tuberosum). Overall, the exact way that ABA is integrated in the biogenic control of chloroplast morphogenesis is still an open question.Besides signals from mature chloroplasts in response to environmental constraints , retrograde signaling pathways also act during chloroplast development to control transcription of nucleus-encoded genes related to photosynthesis . These biogenic signals are mainly studied by inhibition of carotenoid biosynthesis with norflourazon and by inhibition of the translation machinery in plastids with lincomycin (ALTERNATIVE OXIDASE 1A (AOX1a) after perturbation of mitochondrial electron transport (REGULATORS OF AOX1A (RAOs) genes and thereby components of mitochondrial retrograde signaling. One of the first identified RAOs is the DREB transcription factor ABI4 , a mitochondria-localized protein involved in mitochondrial retrograde signaling  and dinor-12-oxo-10,15(Z)-phytodienoic acid (dnOPDA), are made in chloroplasts (cis-OPDA is α-linolenic acid (C18:3) released from the chloroplast galactolipids monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG). By contrast, dnOPDA is synthesized via a parallel hexadecanoid pathway from the less abundant 16:3 fatty acid roughanic acid  (Jasmonate is a collective term used for jasmonic acid (JA) and a diverse set of precursors and derivatives. They regulate a plethora of processes ranging from defense against stresses to the regulation of plant growth and development -hydroperoxylinenic acid (13-HPOT), also gives rise to various volatiles important in the odors of fruits and vegetables  was down-regulated by jasmonate in a COI1-dependent manner  lyase B. This leads to an increase in thiol metabolites and the cellular redox potential  acts as a ‘damage-associated molecular pattern’ signal and is associated with a number of secondary messengers. Wounding, which induces ATP release, also induces expression of jasmonate-dependent genes. 2O2 production upon herbivory  plays a central regulatory role throughout the whole plant life cycle from germination to integration of environmental changes to fruit ripening. Given its importance, the role of ethylene in plant growth, its biosynthesis pathways, and its downstream signaling pathways have been under constant investigation since its discovery as a plant growth factor by S-adenosyl-l-methionine (SAM) to 1-aminocyclopropane 1-carboxylate (ACC) and 5ʹ-methylthioadenosine (MTA) . Cystathionine β-lyase (CBL) then catalyses the conversion of cystathionine to homocysteine. Finally, methionine synthase (MS) methylates homocysteine, using 5-methyltetrahydrofolate as a methyl donor, into methionine  converts f oxygen . Howeverthionine , which ilutescent 2 loss of function mutant that displays slower chloroplast development and altered fruit ripening compared with wild type. Lutescent 2 was found to be the ortholog of the Arabidopsis EGY1 gene, which encodes a plastid thylakoid membrane-localized zinc-dependent M50 type metalloprotease  subunit NDH-M, revealed a delay in the onset of the fruit ripening phenotype as well as ~50% reduced ethylene levels emitted from fruits compared with the wild type  has an anterograde function in chloroplast biogenesis itself. mt) leads to a transient ROS burst within the first 60 min of triggered stress in Arabidopsis. That correlated with an increased activating phosphorylation of MITOGEN ACTIVATED PROTEIN KINASE 6 (MAPK6) in the same time frame. MAPK6 activity led to an increase in transcription of ACS6, thereby activating the ethylene response leading to a part of the transcriptional changes observed for the initially elicited UPRmt, ultimately restoring mitochondrial protein homeostasis  (ETHYLENE-DEPENDENT GRAVOTROPISM-DEFICIENT and YELLOW-GREEN1 (EGY1) gene encodes a thylakoid membrane-localized protease involved in chloroplast development in leaf mesophyll cells that was isolated as an ethylene-dependent gravitropism mutant  and phenylalanine ammonia-lyase (PAL) pathway to synthesize SA, both starting from chorismate precursor  Fig. 6)Fig. 6). ENHANCED DISEASE SUSCEPTIBILITY1 (EDS1) (ENHANCED DISEASE SUSCEPTIBILITY5 (EDS5)   with different structures, localizations, and functions have been identified (From the data published up to now, it can be concluded that phytohormone and organellar signaling works in concert at various levels. However, the studies discussed in this review also make clear that the connections between organellar and phytohormone signaling depend on complex factors such as tissue type, developmental status, and environmental stress conditions (


Korean government has established various policies to counter the low fertility rate since the mid-2000s, but it still has the lowest fertility rate among OECD member countries. This study investigated the relationship between changes in the Korean fertility rate and policies to encourage fertility.This study utilized data of the total fertility rate of 250 local governments between 2014 and 2018, and a casebook of local government birth promotion policies. The dependent variable was fertility rate, and the independent variable was fertility promotion policy. Using SPSS 25.0 and M-plus 8.0 programs, descriptive statistical analysis and latent growth modeling were conducted. An unconditional quadratic function change model was selected as a final model of this study.The average of the initial fertility rate and the linear rate of change in the Korean fertility rate, and the rate of change of the quadratic function were all statistically significant, meaning that the fertility rate of decline increases over time. Also, the linear rate of change and the quadratic function change rate were significant, showing significant differences in the initial level and rate of change of the fertility rate between local governments. Among fertility policies, only the in-kind policy had a significant effect on the initial value of the fertility rate, meaning that the higher the number of in-kind policies, the higher the fertility rate.This study is crucial as it examined the changes in the fertility rate of Korean local governments as well as the policy factors affecting the fertility rate at a quantitative level. There is a concern about low fertility in many countries. Since the 2000s, it has been predicted that the low fertility rate would become a global crisis and will reach the level of ultralow fertility within the next 20 years . Each coWhile the fertility rate started to decline before the 1990s in many OECD countries, this issue started in the mid2000s in South Korea, and the fertility rate has declined rapidly . It is dThe main factors leading to low fertility in South Korea and background causes are discussed as follows. The first factor is socioeconomic. It is discovered that the fertility rate decreases as women’s education rapidly increases , 8. As wLow fertility is a critical issue worldwide. Some scholars argue that the result of low fertility is due to liberalism and neoliberalism in common. In other words, the wave of freedom is affecting the quality of life of individuals, and a lot of effort is put into forming and maintaining a family in a capitalist society, making it difficult to form a family . In addiHowever, most previous studies have dealt with the problem of low fertility in a system, women’s labor force –21, and South Korea has introduced various policies not only at the central government but also at the local government level to respond to the low fertility problem. The central and local governments are pursuing the direction of the fertility promotion policy by focusing on changes in values and the environment to improve individuals’ quality of life. However, previous studies have not considered low fertility policies comprehensively. They have focused on some areas such as childbirth, childcare, and work-family balance, fragmentarily examining them in relation to low fertility .Various policies have emerged to overcome the low fertility rate and encourage childbirth in South Korea over the past decade. Though “The 4th Basic Plan for Low Fertility and Aging Society (2021–2025)” is implemented three times, the phenomenon of extremely low fertility continues to intensify, and the natural decline of the population and demographic onus (a phenomenon in which economic growth is delayed due to a decrease in the working-age population of ages 15 to 64) has become more severe. Accordingly, the necessity of a new vision arose.South Korea’s low fertility-related policies can be divided into cash policy, in-kind policy, voucher policy, service policy, and education and public relations policy. As a cash policy, there is a policy to support the cost of treatment for infertile couples and child allowance paid to all children under the age of 7. As an in-kind policy, various policies have been formulated. For example, diapers and formula are provided to low-income families, and iron and folic acid supplements are provided to pregnant women registered to public health centers. Voucher policies include a maternal/newborn health management support project and a coupon issuance policy to support the cost of ultrasound examinations for pregnant women. As for service policy, education and publicity, health-related medical support, childbirth preparation classes, pregnant women’s classes, and various campaigns are being carried out.However, there are insufficient previous studies examining these policies’ effectiveness in reducing the fertility rate. While some previous studies verified the effectiveness of major policies related to low fertility, most studies were carried out cross-sectionally , 31 and This study utilized the longitudinal data from 2014 to 2018 on the total fertility rate of 250 local governments  provided by Statistics Korea to estimate changes in the fertility rate. Statistics Korea is the central administrative agency of South Korea, and it is in charge of setting standards for statistics, census, and various statistics. The total fertility rate provided by Statistics Korea is a compilation of birth data reported to each local government in accordance with the Statistics Act and the Act on Family Relations Registration.The fertility rate used in this study is the total fertility rate (TFR). TFR is an index indicating the average number of births that one woman of childbearing age (ages 15–49) is expected to have during her lifetime. It is the sum of fertility rates by age and a representative indicator of the level of fertility.As for the fertility promotion policy, the number of each policy is used by dividing the fertility incentive policies implemented by each local government into cash, in-kind, vouchers, services, education and public relations through the Ministry of Health and Welfare’s 2014 Local Government Birth Encouragement Policy Casebook. In the case of cash, congratulatory cash for childbirth, child support expenses, childcare allowance, treatment expense for infertility couples, postpartum care expenses, and local currency were set, and in-kind items were set as birth celebration gifts, safety kits for infants, nutritional supplements, and diapers. Vouchers were set for the maternal and newborn babysitting business, ultrasound coupons, and fetal malformation test coupons, and services were set for rental of toys and books, use of facilities and programs, helper support, and free medical treatment. Education and publicity were set up as childbirth preparation classes, breastfeeding classes, childcare classes, pregnant women classes, infant health classes, events, festivals, and campaigns.The analysis method and procedure for solving the research problem are as follows. SPSS 25.0 and M-plus 8.0 programs were used for data handling and model analysis. First, descriptive statistical analysis was conducted to identify the characteristics of major variables. Second, latent growth modeling was conducted to estimate changes in the overall fertility rate and to verify the relationship between fertility policies. To determine the model fit, TLI (Tucker-Lewis Index), CFI (Comparative Fit Index), and RMSEA (Root Mean Square Error of Approximation) were used. The model is suitable if the CFI and TLI are 0.9 or more  and the The potential growth model conducted in this study is a model that verifies the longitudinal change flow by identifying the amount of change in the variable over time . The potDescriptive statistical analysis was carried out to confirm the overall characteristics of major variables Table . First, In this study, the model was analyzed in two stages. In the first stage, the initial value and change rate were estimated through unconditional model analysis, and in the second stage, the relationship between the change in the fertility rate and fertility incentive policies in South Korea was examined based on the initial value and change rate obtained in the first stage through conditional model analysis.p < .001), CFI = 0.949, TLI = 0.915, RMSEA = 0.080. The fit confirms that the quadratic function change model is more suitable for explaining the change in the fertility rate better than the no-change and linear change models. Thus, the quadratic function change model was adopted, and it was confirmed that the rate of change in the fertility rate increased or decreased differently as the quadratic curve over time.Before proceeding with the conditional model analysis, an unconditional model analysis was performed to understand the change in the fertility rate. In order to identify the optimal change pattern through the unconditional model, the no-change model, the linear change model, and the quadratic function change model were analyzed, respectively Table . The fitp < .001), and the linear rate of change in the Korean fertility rate was 0.016 (p. <0.05), and the rate of change of the quadratic function was − 0.020 (p < .001) (Table p < .001), the linear rate of change 0.004 (p < .05), and the quadratic function change rate of 0.003 (p < .05). This shows that there are significant differences in the initial level and rate of change of the fertility rate between local governments.According to the analysis result of the finally selected unconditional quadratic function change model, the average of the initial fertility rate (fertility rate in 2014) was 1.316 (p < .001), CFI = 0.952, TLI = 0.906, and RMSEA = 0.089.In the conditional model analysis, the effect of the fertility promotion policy on the initial value and change rate of the fertility rate was examined. As a result of conditional model fit analysis, it was found that there was no problem in model analysis with χ2 = 111.122 (p < .001)  Table . In otheDuring the ultra-low fertility crisis facing the world, South Korea is currently unable to escape from the lowest fertility rate. Since the mid-2000s, the Republic of South Korea has established comprehensive government-level countermeasures against low fertility and mobilized enormous financial resources to implement various policies to encourage childbirth. However, the fertility rate is currently the only OECD member country with a total fertility rate of zero rather than rebounding . Thus, tAs a result of the analysis, the change in the fertility rate in South Korea shows changes over time, and the shape of the trajectory is close to a curved trajectory in the form of a quadratic function. In other words, as time goes by, the rate of decline increases, indicating that the problem of declining fertility rates is very serious.In addition, it was found that the fertility promotion policies proposed by South Korea did not have a positive effect on the fertility rate and did not slow the rate of decline in the fertility rate. South Korea has been implementing related policies until now through the establishment of the Basic Plan for Low Fertility and Aging Society in 2006, and the number of related policies, which was 1,306 in 2014, at the time of this study, reached 1,747 in 2018 , 37. BasOn the other hand, it was found that the in-kind policy had a temporary effect on the fertility rate. In the case of in-kind policies, it is mainly implemented during pregnancy and childbirth. Typically, nutritional supplements  are provided during the pregnancy phase, and after childbirth, childbirth celebration supplies that can help with the provision of early childbirth supplies are in progress. Compared to other policies that provide different types of support according to the order of births, it can be inferred that the results are derived from the fact that there are many contents that correspond to the support criteria for pregnant and childbirth families. For the lasting effect of this spot policy, a customized approach seems to be necessary considering more diverse life cycles.However, it is unreasonable to conclude this study as a failure of South Korea’s fertility policy. It is because even if the effect of a policy is positive, it can be offset by other factors . In addiSeveral studies examining the effectiveness of South Korea’s fertility policies have determined that related policies have a positive effect on marital fertility but have a negative correlation with the marriage rates of single individuals. Pointing out that factors may differ, it is impossible to say that the policy to encourage childbirth is ineffective . In addiIn this regard, this study has obvious limitations. This study looked at quantitatively by measuring only numbers without considering the characteristics of each policy. A broader discussion will be possible if we consider the qualitative aspects of the policy, such as considering the various characteristics of the policy and examining the total fertility rate in detail. Nevertheless, this study is meaningful in that it explores the relationship by examining changes in the fertility rate and policy factors at a quantitative level.In this study, we examined the changes in ts'he Korean fertility rate and the relationship between changes in the fertility rate and policies to encourage fertility. Consequently, the change in the fertility rate in South Korea showed a rapid increase in the rate of decline over time, and it was found that the fertility incentive policy did not affect both the fertility rate and the rate at which the fertility rate decreased. It shows that the quantitative expansion of the fertility promotion policy, which has been overheated in local governments, does not have a positive effect on the fertility rate in the short and long term and does not reflect the region’s characteristics. For the future studies, an impactful policy with a more precise purpose and target is necessary rather than quantitative expansion.