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Summary Objective We aimed to investigate mechano-sensitivity at the afferent nerve fibers projecting to degenerated intervertebral disc (IVD) and nociceptive behaviour in a rat model of low back pain (LBP). Design Animal model with LBP was established by lumbar 4/5 IVD puncture and nucleus pulposus aspiration. In vivo single nerve recordings (n = 121) were introduced to measure discharge frequency at the afferent nerve fiber innervating the IVD during mechanical stimulations (von Frey filament or intradiscal pressure). Nerve growth factor (NGF) expression levels in the IVD (n = 20) were assessed by Western blot. LBP-related behaviour (n = 22) was assessed by measuring changes in rearing, mechanical paw-withdrawal threshold, and dynamic weight bearing in a freely walking rat. Inhibitory effect of morphine on the neuronal excitability (n = 19) and painful behaviour (n = 28) was also assessed. Results Compared to those with sham or naive IVD, animal group with degenerated IVD displayed the sensitized neuronal responses and painful behaviour, with hyperexcitability of the afferent nerve fibers in any range of mechanical stimulations (von Frey filament stimulation; 1, 2, and 26 g; intradiscal pressure, 1,500–3,000 mm Hg), strong upregulation of NGF (200–250 % increase), and LBP-like behaviour such as failure of rearing, front limbs-dependent walking pattern, and hypersensitivity in hind-paws. However, the neuronal hyperexcitability and pain behaviour were attenuated after local (30 μM) or systemic (3 mg kg−1) morphine administration. Conclusions Our study suggests that enhanced mechano-sensitivity at the afferent nerve fiber innervating degenerated IVD is deeply correlated with LBP development, which supports the hypothesis that hyperexcited responses at the nerve fibers represent a decisive source of LBP.

While initial encoding of contextual memories involves the strengthening of hippocampal circuits, these memories progressively mature to stabilized forms in neocortex and become less hippocampus dependent. Although it has been proposed that long-term storage of contextual memories may involve enduring synaptic changes in neocortical circuits, synaptic substrates of remote contextual memories have been elusive. Here we demonstrate that the consolidation of remote contextual fear memories in mice correlated with progressive strengthening of excitatory connections between prefrontal cortical (PFC) engram neurons active during learning and reactivated during remote memory recall, whereas the extinction of remote memories weakened those synapses. This synapse-specific plasticity was CREB-dependent and required sustained hippocampal signals, which the retrosplenial cortex could convey to PFC. Moreover, PFC engram neurons were strongly connected to other PFC neurons recruited during remote memory recall. Our study suggests that progressive and synapse-specific strengthening of PFC circuits can contribute to long-term storage of contextual memories.

ObjectiveDrug injections and surgery are popular treatments for knee joint osteoarthritis. However, these treatments are invasive, and new noninvasive treatments with similar or better efficacy are needed. Here, we evaluated the application of 4.4 MHz of pulsed radiofrequency (PRF) as a new treatment.MethodsAcute arthritis was induced by injection of carrageenan into the intra-articular space of the knee in male rats. At 4.5 hours after arthritis induction, PRF with the treatment protocol of three seconds on and off was applied to the affected knee joint for 20 minutes. The changes in pain behavior were evaluated by comparing the peak weight load values of both hind paws at pretreatment and four, six, seven, eight, and 24 hours after treatment. And we also used Western blotting and immunohistochemistry to measure the inflammatory changes in the synovial membrane of the inflamed knee.ResultsWe found that the 20-minute application of PRF with the treatment protocol significantly recovered the weight load reduction at six-, seven-, and eight-hour time points after carrageenan injection. COX-2 and IL-1β levels were significantly reduced in the inflamed rats after PRF application at six and eight hours post–carrageenan injection. Immunohistochemistry showed that PRF significantly reduced inflammatory cell infiltration at six hours post–carrageenan injection.Conclusions. Our results indicate that noninvasive PRF application inhibited pain-related behavior and decreased inflammatory cytokine expression in the inflamed knee joints of rats. Accordingly, PRF application can serve as a potential therapeutic treatment to relieve pain associated with peripheral joint/tissue damage or inflammation.

Introduction Lengthened immobilization may prevent muscle shortening, and help maintain normal muscle length. However, its apoptotic effects remain unclear. We evaluated the effects of long-term immobilization on apoptotic proteins. Methods Rat soleus muscles were immobilized by casting in a neutral (NEUT) or lengthened (LENG) position for 21 days. We evaluated dynamic weight load and muscle atrophy following the 21-day period using hematoxylin and eosin staining. We measured Bax (pro-apoptotic Bcl-2 family member), MyoD (myogenic differentiation factor D), MYH (myosin heavy chain), and cleaved poly(ADP-ribose)polymerase levels and examined apoptotic nucleus expression. Results Decreased dynamic weight load and muscle atrophy changes were observed in LENG. Both NEUT and LENG showed significantly reduced levels of MYH. LENG showed a significant increase in Bax and MyoD expression as well as in the number of apoptotic nuclei. Conclusions Long-term lengthened immobilization may increase apoptotic changes and decrease muscle formation proteins in muscle. Muscle Nerve 59:263-269, 2019.

Development of plant tissues is dependent on numerous factors, including hormone activity, signaling, cell division, and elongation. In plants, Defective Entry into Mitosis 1 (Dim1) homologs are recognized as pivotal in leaf senescence and progress of normal growth, but their role in rice has not been functionally characterized. The findings presented in this paper suggest that OsDim1 is important in early seedling development, pollen tube elongation, and impacts rice yield components. The gene is expressed in the scutellum, endosperm, embryonic root, shoot, pollen grains and tubes, as well as in several organs of the rice flower. According to the present study findings, RNAi mediated knockdown of OsDim1 resulted in phytohormonal imbalance, reduced amylase activity, affected differentiation of embryonic root elongation zone tissues, suppressed embryonic root and shoot growth, and impaired pollen tube elongation. In contrast, overexpression of OsDim1 showed significant growth in embryonic roots and shoots, while it increased culm length, total number of tillers per plant, seed setting rate, and total number of grains per panicle compared to its wild type line. In summary, we propose OsDim1 plays an important role in seedling growth and pollen tube elongation, and has pleiotropic effects on reproductive tissues.

The dim1+ gene family is essential for G2/M transition during mitosis and encodes a small nuclear ribonucleoprotein that functions in the mRNA splicing machinery of eukaryotes. However, the plant homolog of DIM1 gene has not been defined yet. Here, we identified a gene named GmDim1 positioned on chromosome 9 of soybean (Glycine max (L.) Merr.) with 80% homology to other eukaryotic dim1+ family genes. A domain of soybean DIM1 protein was primarily conserved with U5 snRNP protein family and secondarily aligned with mitotic DIM1 protein family. The GmDim1 gene was expressed constitutively in all soybean organs. The transgenic Arabidopsis thaliana (L.) plants overexpressing GmDim1 showed early flowering and stem elongation, produced multiple shoots and continued flowering after the post-flowering stage. DIM1 proteins transiently expressed in onion cells were localized in the nucleus with dense deposition in the nucleolus. Therefore, we propose that the soybean GmDim1 gene is a component of plant U5 snRNP involved in mRNA splicing and normal progress of plant growth.

The systemic administration of opioids can be used for their strong analgesic effect. However, extensive activation of opioid receptors (ORs) beyond the targeted tissue can cause dysphoria, pruritus, and constipation. Therefore, selective activation of peripheral ORs present in the afferent fibers of the targeted tissue can be considered a superior strategy in opioid analgesia to avoid potential adverse effects. The purpose of this study was to clarify the role of peripheral kappa opioid receptors (kORs) in arthritic pain for the possible use of peripheral ORs as a target in anti-nociceptive therapy. We administered U50488 or nor-BNI/DIPPA, a selective agonist or antagonist of kOR, respectively into arthritic rat knee joints induced using 1% carrageenan. After the injection of U50488 or U50488 with nor-BNI or DIPPA into the inflamed knee joint, we evaluated nociceptive behavior as indicated by reduced weight-bearing on the ipsilateral limbs of the rat and recorded the activity of mechanosensitive afferents (MSA). In the inflamed knee joint, the intra-articular application of 1μM, 10nM, or 0.1nM U50488 resulted in a significant reduction in nociceptive behavior. In addition, 1μM and 10nM U50488 decreased MSA activity. However, in a non-inflamed knee joint, 1μM U50488 had no effect on MSA activity. Additionally, intra-articular pretreatment with 20μM nor-BNI or 10μM DIPPA significantly blocked the inhibitory effects of 1μM U50488 on nociceptive behavior and MSA activity in the inflamed knee joint. These results implicate that peripheral kORs can contribute to anti-nociceptive processing in an inflamed knee joint.

Chamaecyparis obtusa essential oil (COE) has been widely used to treat allergic diseases and was suggested to exert anti-inflammatory, antioxidant, and antimicrobial effects. This study evaluated the effects of COE on pain-related behavior and pro-inflammatory cytokines in rats with carrageenan (CGN)-induced arthritis. Reduced dynamic weight load on inflamed joint in voluntarily walking rats was used as the behavior test for arthritic pain; 10% COE-treated group was significantly attenuated pain (6–8 h post-CGN injection) compared to VEH (mineral oil)-treated group. In addition, the protein levels of interleukin (IL)-1β, tumor necrosis factor-α, IL-6 (6–8 h), and cyclooxygenase (COX)-2 (8 h) within the synovial membrane, as well as IL-1β, COX-2 (6–8 h), and IL-6 (5–7 h) within the meniscus, of 10% COE-treated group were significantly reduced. The current results implicate that COE has anti-inflammatory and anti-nociceptive effects on arthritis in rats.

Peripheral group III metabotropic glutamate receptors (mGluRs) function to modulate pain signaling in inflammatory states. Here, we established in vivo experimental settings, including dynamic weight bearing test and in vivo single nerve recording, to elucidate how the group III mGluRs contribute to inhibiting pain transmission at the peripheral sensory nerve terminal in inflammatory states (1 and 3 days) elicited by Complete Freund’s Adjuvant (CFA). As a result, CFA-induced nociceptive behaviors were significantly alleviated after administration of 100 and 200 μM L-AP4 (l-2-amino-4-phosphonobutylate; group III mGluR agonist). In addition, neuronal discharges evoked by 6- and 26-g von Frey filaments at the nerve significantly decreased after administration of 200 μM L-AP4. However, this event was not observed in non-inflammatory state. These results suggest that the group III mGluRs negatively regulate nociceptive behavior and pain transmission by lessening neuronal firing rates at the peripheral nerve in inflammation.

Isolation stress is a major risk factor for neuropsychiatric disorders such as depressive and anxiety disorders. However, the molecular mechanisms underlying isolation-induced neuropsychiatric disorders remain elusive. In the present study, we investigated the subcellular mechanisms by which long-term isolation elicits depression and anxiety-related behaviors in mice. First, we found that long-term isolation induced depression-related behaviors in the forced swimming test (FST) and the sucrose preference test, as well as anxiety-related behaviors in the elevated zero maze test (EZMT) and the open field test. Next, we showed that intracentral amygdala (CeA) injection of oxytocin (OXT), but not intracerebroventricular injection, attenuated isolation-induced depression and anxiety-related behaviors via oxytocin receptor (OXTR), not vasopressin-1a receptor (V1aR), in the FST and EZMT, respectively. Quantitative real-time polymerase chain reaction analysis revealed that after 5 weeks of isolation, mRNA transcription of OXTR in the CeA, but not that of V1aR, significantly decreased, whereas OXT and vasopressin mRNA transcription in the paraventricular nucleus of hypothalamus did not change significantly. Whole-cell patch clamping of acute brain slices demonstrated that the frequency of miniature inhibitory postsynaptic currents (mIPSCs) in CeA neurons, but not their amplitude, was lower in isolated mice than in group-housed mice. Notably, OXT treatment increased the mIPSC frequency in the CeA neurons, but to a lesser extent in the case of isolated mice than in that of group-housed mice via OXTR. Taken together, our findings suggest that long-term isolation down-regulates OXTR mRNA transcription and diminishes OXT-induced inhibitory synaptic transmission in the CeA and may contribute to the development of depression and anxiety-related behaviors in isolated mice through the enhancement of CeA activity.

Genetic variation in wild soybean (Glycine sojaSieb. and Zucc.) is a valuable resource for crop improvement efforts. Soybean is believed to have originated from China, Korea, and Japan, but little is known about the diversity or evolution of Korean wild soybean. Therefore, in this study, we evaluated the genetic diversity and population structure of 733G. sojaaccessions collected in Korea using 21 simple sequence repeat (SSR) markers. The SSR loci produced 539 alleles (25.7 per locus) with a mean genetic diversity of 0.882 in these accessions. Rare alleles, those with a frequency of less than 5%, represented 75% of the total number. This collection was divided into two populations based on the principal coordinate analysis. Accessions from population 1 were distributed throughout the country, whereas most of the accessions from population 2 were distributed on the western side of the Taebaek and Sobaek mountains. The KoreanG. sojacollection evaluated in this study should provide useful background information for allele mining approach and breeding programmes to introgress alleles into the cultivated soybean (G. max(L). Merr.) from wild soybean.

Mungbean sprout rot is one of the most serious problems of the commercial mungbean sprout industry. In this study, 70 strains of mungbean sprout rot pathogens were isolated from rotten sprouts at different time intervals. The pathogenicity of the isolated pathogens was tested. The highly pathogenic strain (YV-St-033) was identified as Pseudomonas sp. by 16S rRNA gene sequencing. In phylogenetic analysis, the YV-St-033 strain was grouped with P. mosselii, P. putita, P. fluorescens, P. entomophila, and P. lecoglossicida. The results of the 16S rRNA gene sequence analysis revealed that the YV-St-033 strain shared the highest sequence identity (more than 99%) with the P. mosselii R10 strain. The mungbean lines of Yeungnam University germplasm were screened against the YV-St-033 strain. Based on the growth rate of the sprouts after 3 days of inoculation with the pathogen, the YV148 line was highly resistant to the pathogen. The remaining lines were either partially or fully infected. The highly resistant line YV 148 is suitable for future breeding programs due to their thin sprouts and fast growing nature.

On screening pathogen-resistant soybean, we identified a WRKY type transcription factor named a Glycine max WRKY1 (GmWRKY1). Expression of GmWRKY1 gene was induced in the soybean sprout by Pseudomonas infection. The GmWRKY1 was expressed in all of the tissues with high levels in stems, leaves and developing seeds. The protein Gm WRKY1 contains highly conserved two WRKY DNA-binding domains having two zinc-finger motif () in its N-terminal and C-terminal amino acid sequences. In electrophoresis mobility shift assay, the GmWRKY1 protein bound specifically to W-box elements in the promoters of defense related genes. These results demonstrated that GmWRKY1 is one of the soybean WRKY family genes and the plant-specific transcription factors for defense processes.

Control microbial contamination in pathogens to soy sprouts has always been highly concerned in soybean sprout industries because the soybean sprouts are consumed largely as a nutritious fresh vegetable around the world. However, pathogens in soy sprouts are little known. Here, we isolated a strain of Pseudomonas sp. SN239 that caused severer symptoms in sprouts of many soybean cultivars. In phylogenetic relationships using 16S ribosomal RNA sequences of the Pseudomonas species, the identified Pseudomonas sp. SN239 was grouped with P. putita, P. plecoglossicida, P. monteilii and P. mevalonii. Thus, the bacterial strain SN239 might be a newly identified Pseudomonas species which closely related to P. putida. Furthermore, we found that a Korean indigenous soybean (Glycine max) cultivar YNPCSS3-19 has strong resistance against the Pseudomonas sp. SN239.

In Korea, a local soybean (Glycine max) genotype 56l. was found to be strongly resistant to a virulent bacterial strain of a Pseudomonas sp. SN239. Specific genes involved in the resistance of the soybean genotype 561 were identified and the pattern of gene expression against the Pseudomonas infection was analyzed using differential-display reverse transcription PCR (DDRT-PCR). More than 126 cDNA fragments representing mRNAs were induced within 48 hours of bacteria inoculation. Among them, 28 cDNA fragments were cloned and sequenced. Twelve differentially displayed clones with open reading frames had unknown functions. Sixteen selected cDNA clones were homologous to known genes of other organisms. Some of the identified cDNAs were pathogenesis-related (PR) genes and PR-like genes. These cDNAs included a putative calmodulin-binding protein; an endo-l,3-1,4--D-glucanase; a -1,3-endoglucanase; a -1,3-exoglucanase; a phytochelatin synthetase-like gene; a thiol protease; a cycloartenol synthase; and a putative receptor-like serine/threonine protein kinase. Among them, four genes were found to be putative PR genes induced significantly by the Pseudomonas infection. These included a calmodulin-binding protein gene, a -1,3-endoglucanase gene, a receptor-like serine/threonine protein kinase gene, and pS321 (unknown function). These results suggest that the differentially expressed genes may mediate the strong resistance of soybean 561 to the strain SN239 of Pseudomonas sp.

This study sought to evaluate whether peripheral group II and III metabotropic glutamate receptors (mGluRs) in the knee joint have inhibitory effects on carrageenan-induced nociceptive behavior. To this end, changes in weight load and hind paw withdrawal threshold were measured in rats following the administration of specific peripheral group II and III mGluR agonists 30min before (induction phase) and 4h after (maintenance phase) the injection of carrageenan (1%, 50μl). During the induction phase of arthritic pain, a significant recovery of reduced weight load occurred after the administration of 500μM APDC ((2R, 4R)-4-aminopyrrolidine-2,4-dicarboxylate; group II agonist) and 100 and 500μM L-AP4 (l-2-amino-4-phosphonobutylate; group III agonist). Similarly, 100 and 500μM APDC and 500μM L-AP4 significantly reduced mechanical hyperalgesia during the induction phase. In the maintenance phase, APDC at all doses (10, 100 and 500μM) and 100 and 500μM L-AP4 significantly counteracted the reduction in weight load, and APDC and L-AP4 at all doses (10, 100 and 500μM) inhibited mechanical hyperalgesia. The current study suggests that peripheral group II and III mGluRs in the knee joint negatively modulates nociceptive behavior during both the induction and maintenance phases of carrageenan-induced arthritic pain.

ity, which led to very mild cartilage degeneration. The positive correlation of cartilage degeneration with the degree of instability supports the empirically-plausible concept that joints with higher degrees of joint instability are at a higher risk of degeneraation. High incidence of severe cartilage degeneration in the medial femoral surface in the total ACLT knees may be associated with accompanying medial meniscus tear. To introduce more reproducible, sub-critical level cartilage degeneration, the degree of instability may need to be higher, but should be within a range not to cause medial meniscus tear.