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Functional Dissection of Glutamatergic and GABAergic Neurons in the Bed Nucleus of the Stria Terminalis

  • Kim, Seong-Rae;Kim, Sung-Yon
    • Molecules and Cells
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    • v.44 no.2
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    • pp.63-67
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    • 2021
  • The bed nucleus of the stria terminalis (BNST)-a key part of the extended amygdala-has been implicated in the regulation of diverse behavioral states, ranging from anxiety and reward processing to feeding behavior. Among the host of distinct types of neurons within the BNST, recent investigations employing cell type- and projection-specific circuit dissection techniques (such as optogenetics, chemogenetics, deep-brain calcium imaging, and the genetic and viral methods for targeting specific types of cells) have highlighted the key roles of glutamatergic and GABAergic neurons and their axonal projections. As anticipated from their primary roles in excitatory and inhibitory neurotransmission, these studies established that the glutamatergic and GABAergic subpopulations of the BNST oppositely regulate diverse behavioral states. At the same time, these studies have also revealed unexpected functional specificity and heterogeneity within each subpopulation. In this Minireview, we introduce the body of studies that investigated the function of glutamatergic and GABAergic BNST neurons and their circuits. We also discuss unresolved questions and future directions for a more complete understanding of the cellular diversity and functional heterogeneity within the BNST.

Erythrocyte Sedimentation Rate: Its Determinants and Relationship with Risk Factors Involved in Ischemic Stroke

  • Kaur, Kirandeep;Kaur, Amandeep;Kaur, Anupam
    • Korean Journal of Clinical Laboratory Science
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    • v.54 no.1
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    • pp.1-8
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    • 2022
  • Erythrocyte sedimentation rate (ESR) evaluation is a useful tool for monitoring disease activity in various inflammatory and non-inflammatory conditions. ESR is known to be influenced by a multitude of confounding factors. The present study aimed to assess the possible determinants of the ESR and its relationship with various risk factors involved in ischemic stroke. ESR and other hematological and biochemical parameters were investigated in 163 ischemic stroke patients (107 males and 56 females) selected based on imaging techniques including magnetic resonance imaging (MRI) or computed tomography (CT) scans. Statistical analysis was performed using the SPSS 16.0 software. Linear regression analysis showed a significant inverse relationship of hemoglobin (Hb) and hematocrit or packed cell volume (PCV) (P<0.001 for females; P<0.01 for males) with the ESR. It was observed that the red blood cell (RBC) count was not strongly correlated with the ESR (P<0.05 for both males and females). It was also observed that sex significantly affected the variables determining the ESR levels, whereas age had no effect. Gender differences were also observed with respect to Hb, RBC, PCV, mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and ESR. The possible determinants of higher ESR levels in ischemic stroke may be sex, Hb, hematocrit, and RBC count, but the role of other clinical and laboratory parameters cannot be underestimated.

Characterization and RT-PCR Detection of dsRNA Mycoviruses from the Oyster Mushroom, Pleurotus ostreatus

  • Seo, Jang-Kyun;Lim, Won-Seok;Jeong, Ji-Hye;Yoo, Young-Bok;Yie, Se-Won;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.20 no.3
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    • pp.200-205
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    • 2004
  • The partial nucleotide sequences of the genomic dsRNA mycovirus infecting Pleurotus ostreatus isolates ASI2223 and Suhan were determined and compared with those of mycoviruses belonging to partitiviruses and totiviruses. Partial nucleotide sequences of the purified dsRNA from ASI2223 and Suhan showed RNA-dependent RNA polymerase sequences that are closely related to those of partitiviruses, including Fusarium poae virus 1, Fusarium solani virus, Rhizoctoniasolani virus, Discula destructiva virus 2, and Oyster mushroom isometric virus 2. Specific primers were designed for RT-PCR detection of dsRNA viruses from the P. ostreatus isolate ASI2223 and Suhan. Two virus specific primer sets were found to specifically detect each virus among six sets of designed oligonucleotide primers. Collectively, these results suggest that dsRNA mycoviruses from P. ostreatus isolates ASI2223 and Suhan belong to the family Partitiviridae, although, they are not the same virus species. Our results also suggest that these virus-specific primer sets can be employed for the specific detection of each viral sequence in infected tissues.

Molecular Characterization of a dsRNA Mycovirus, Fusarium graminearum Virus-DK21, which Is Phylogenetically Related to Hypoviruses but Has a Genome Organization and Gene Expression Strategy Resembling Those of Plant Potex-like Viruses

  • Kwon, Sun-Jung;Lim, Won-Seok;Park, Sang-Ho;Park, Mi-Ri;Kim, Kook-Hyung
    • Molecules and Cells
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    • v.23 no.3
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    • pp.304-315
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    • 2007
  • Fusarium graminearum causes a serious scab disease of small grains in Korea. The nucleotide sequence of the genomic RNA of a double-stranded RNA (dsRNA) virus, Fusarium graminearum virus-DK21 (FgV-DK21), from F. graminearum strain DK21, which is associated with hypovirulence in F. graminearum, was determined and compared to the genome sequences of other mycoviruses, including Cryponectria hypoviruses. The FgV-DK21 dsRNA consists of 6,624 nucleotides, excluding the 3'-terminal poly(A) tail. The viral genome has 53- and 46-nucleotide 5' and 3' untranslated regions (UTRs), respectively, and five putative open reading frames. A phylogenetic analysis of the deduced amino acid sequence of ORF1, which encodes a putative RNA-dependent RNA polymerase, and those of other mycoviruses revealed that this organism forms a distinct virus clade with other hypoviruses, and is more distantly related to other mycoviruses (3.8 to 24.0% identity). However, pairwise sequence comparisons of the nucleotide and deduced amino acid sequences of ORFs 2 through 5 revealed no close relationships to other protein sequences currently available in GenBank. Analyses of RNA accumulation by Northern blot and primer extension indicated that these putative gene products are expressed from at least two different subgenomic RNAs (sgRNAs), in contrast to the cases in other hypoviruses. This study suggests the existence of a new, as yet unassigned, genus of mycoviruses that exhibits a potex-like genome organization and sgRNA accumulation.

Elimination of SPFMV from Virus-infected Sweet Potato Plants through Apical Meristem Culture

  • Kim, Young-Seon;Jeong, Jae-Hun;Park, Jong-Suk;Eun, Jong-Seon
    • Plant Resources
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    • v.7 no.3
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    • pp.200-205
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    • 2004
  • Sweet potato infected with a viral disease (SPFMV) showed irregular chlorotic patterns, so called feathering associated with faint or distinct ring spots that have purple-pigmented borders. SPFMV was eliminated from sweet potato plants using meristem tip culture. MS medium supplemented with BAP (2mg/L) and NAA (0.05 mg/L) was used for shoot proliferation and 1/2 MS medium for rooting of the plants. Highest percentage of regenerated plants (60%) was obtained from the optimum size (0.3-0.5mm) meristem tips. Of these, 60% plants were found negative for SPFMV by RT-PCR. Virus detection by RT-PCR was found to be a reliable method. Meristem-tip culture to produce SPFMV-free quality sweet potato and virus detection by RT-PCR is an efficient, time saving and reliable method for production of SPFMV-free tissue culture raised plants.

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Five Newly Collected Turnip Mosaic Virus (TuMV) Isolates from Jeju Island, Korea are Closely Related to Previously Reported Korean TuMV Isolates but Show Distinctive Symptom Development

  • Hu, Wen-Xing;Kim, Byoung-Jo;Kwak, Younghwan;Seo, Eun-Young;Kim, Jung-Kyu;Han, Jae-Yeong;Kim, Ik-Hyun;Lim, Yong Pyo;Cho, In-Sook;Domier, Leslie L;Hammond, John;Lim, Hyoun-Sub
    • The Plant Pathology Journal
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    • v.35 no.4
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    • pp.381-388
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    • 2019
  • For several years, temperatures in the Korean peninsula have gradually increased due to climate change, resulting in a changing environment for growth of crops and vegetables. An associated consequence is that emerging species of insect vector have caused increased viral transmission. In Jeju Island, Korea, occurrences of viral disease have increased. Here, we report characterization of five newly collected turnip mosaic virus (TuMV) isolates named KBJ1, KBJ2, KBJ3, KBJ4 and KBJ5 from a survey on Jeju Island in 2017. Full-length cDNAs of each isolate were cloned into the pJY vector downstream of cauliflower mosaic virus 35S and bacteriophage T7 RNA polymerase promoters. Their fulllength sequences share 98.9-99.9% nucleotide sequence identity and were most closely related to previously reported Korean TuMV isolates. All isolates belonged to the BR group and infected both Chinese cabbage and radish. Four isolates induced very mild symptoms in Nicotiana benthamiana but KBJ5 induced a hypersensitive response. Symptom differences may result from three amino acid differences uniquely present in KBJ5; Gly(382)Asp, Ile(891)Val, and Lys(2522)Glu in P1, P3, and NIb, respectively.

Expression of Human Immunodeficiency Virus Type 1 Tat Proteins in Escherichia coli and Application to Study Tat Functions

  • Park, Jin-Seu;Lee, Han-Gyu;Lee, Yoon;Kang, Young-Hee;Rhim, Hyang-Shuk;Choi, Soo-Young
    • BMB Reports
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    • v.33 no.4
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    • pp.337-343
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    • 2000
  • The human immunodeficiency virus type 1 (HIV-1), transactivator of transcription (Tat), is one of the viral gene products that is essential for HIV-1 replication. The HIV-l Tat protein regulates transcription from an HIV-1 long terminal repeat (LTR) and affects the gene expression of cellular proteins during infection. In order to develop an expression system to overexpress and simply purify HIV-1 Tat proteins, the HIV-1 Tat coding sequences that contain one or two exons were amplified using PCR and cloned into a pET vector, which contains a consecutive stretch of six histidine residues at the amino-terminus. The reconstituted vectors were overexpressed in the E. coli strain and the soluble recombinant proteins were purified to be homogeneity in a single step by $Ni^{+2}-nitrilotriacetic$ acid Sepharose chromatography under nondenaturing conditions. Recombinant HIV-1 Tat proteins were shown to transactivate the HIV-1 LTR promoter in a dose-dependent manner when introduced into mammalian cells. In addition, treatment of human endothelial cells with purified Tat proteins resulted in a significant increase in the level of vascular cell adhesion molecule-1 (VCAM-1) expression. These results indicate that the recombinant HIV-1 Tat proteins are active in transactivating viral and cellular promoters. The expression and purification system described in this study will facilitate in characterizing the biological functions of the Tat proteins.

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Viral Effects of a dsRNA Mycovirus (PoV-ASI2792) on the Vegetative Growth of the Edible Mushroom Pleurotus ostreatus

  • Song, Ha-Yeon;Choi, Hyo-Jin;Jeong, Hansaem;Choi, Dahye;Kim, Dae-Hyuk;Kim, Jung-Mi
    • Mycobiology
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    • v.44 no.4
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    • pp.283-290
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    • 2016
  • A double-stranded RNA (dsRNA) mycovirus was detected in malformed fruiting bodies of Pleurotus ostreatus strain ASI2792, one of bottle cultivated commercial strains of the edible oyster mushroom. The partial RNA-dependent RNA polymerase (RdRp) gene of the P. ostreatus ASI2792 mycovirus (PoV-ASI2792) was cloned, and a cDNA sequences alignment revealed that the sequence was identical to the RdRp gene of a known PoSV found in the P. ostreatus strain. To investigate the symptoms of PoV-ASI2792 infection by comparing the isogenic virus-free P. ostreatus strains with a virus-infected strain, isogenic virus-cured P. ostreatus strains were obtained by the mycelial fragmentation method for virus curing. The absence of virus was verified with gel electrophoresis after dsRNA-specific virus purification and Northern blot analysis using a partial RdRp cDNA of PoV-ASI2792. The growth rate and mycelial dry weight of virus-infected P. ostreatus strain with PoV-ASI2792 mycovirus were compared to those of three virus-free isogenic strains on 10 different media. The virus-cured strains showed distinctly higher mycelial growth rates and dry weights on all kinds of experimental culture media, with at least a 2.2-fold higher mycelial growth rate on mushroom complete media (MCM) and Hamada media, and a 2.7-fold higher mycelial dry weight on MCM and yeast-malt-glucose agar media than those of the virus-infected strain. These results suggest that the infection of PoV mycovirus has a deleterious effect on the vegetative growth of P. ostreatus.

Primary Immunodeficiencies in Children Initially Admitted with Gastrointestinal/Liver Manifestations

  • Murat Cakir ;Nalan Yakici ;Elif Sag ;Gulay Kaya ;Aysenur Bahadir;Alper Han Cebi ;Fazil Orhan
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.26 no.4
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    • pp.201-212
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    • 2023
  • Purpose: The gastrointestinal system is the most commonly affected organ, followed by the lungs, in patients with primary immunodeficiency disease (PID). Hence, it is common for children with PIDs to present with gastrointestinal symptoms. We aimed to analyze the clinical and histopathological findings of patients who were initially admitted to pediatric gastroenterology/hepatology clinics and subsequently diagnosed with PIDs to identify the clinical clues for PIDs. Methods: The demographic, laboratory, and histopathological findings, treatment modality, and outcomes of patients initially admitted to the pediatric gastroenterology/hepatology unit and subsequently diagnosed with PIDs were recorded. Results: The study included 24 patients (58.3% male; median age [range]: 29 [0.5-204] months). Common clinical presentations included chronic diarrhea (n=8), colitis (n=6), acute hepatitis (n=4), and acute liver failure (n=2). The association of autoimmune diseases, development of malignant diseases, and severe progression of viral diseases was observed in 20.8%, 8.3%, and 16.6% of the patients, respectively. Antibody deficiency was predominantly diagnosed in 29.2% of patients, combined immunodeficiency in 20.8%, immune dysregulation in 12.5%, defects in intrinsic and innate immunity in 4.2%, autoinflammatory disorders in 8.3%, and congenital defects of phagocytes in 4.2%. Five patients remained unclassified (20.8%). Conclusion: Patients with PIDs may initially experience gastrointestinal or liver problems. It is recommended that the association of autoimmune or malignant diseases or severe progression of viral diseases provide pediatric gastroenterologists some suspicion of PIDs. After screening using basic laboratory tests, genetic analysis is mandatory for a definitive diagnosis.

Human Immunodeficiency Virus-Infected T Cells Are Selectively Killed by Monoclonal Anti-gp120 Antibody Coupled to Pokeweed Antiviral Protein (섬자리공 유래 항바이러스 단백질과 항체 복합체를 이용한 HIV-1 감염세포의 선택적 제거)

  • Kang, Mi-Ran;Kim, Yoon-Kyu;Hong, Hyo-Jeong;Cho, Myung-Hwan;Shin, Hyung-Sik;Kim, Sun-Young
    • The Journal of Korean Society of Virology
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    • v.28 no.4
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    • pp.383-391
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    • 1998
  • A murine monoclonal antibody (mAb) specific for the envelope glycoprotein gp120 of human immunodeficiency virus type-I (HIV -1) was chemically coupled to pokeweed antiviral protein (PAP) from Phytolacca americana. The immunotoxin was purified by FPLC using S200 colum. The purified immunotoxin efficiently bound to HIV-infected T cells as evidenced by fluorescenceactivated cell sorter analysis. The immunotoxin selectively killed human T lymphoid lines infected with $HIV-1_{IIIB}$ at less than 250 pM of the immunotoxin cells, while PAP or mAb alone did not have any significant effect on infected cells. The uninfected control T cell lines were not affected. Human cells infected with HIV-2 or other HIV-1 strains were not killed, suggesting that the killing depends completely on the antibody used for coupling. These in vitro results suggest that the PAP-mAb conjugate may be used to selectively remove cells expressing viral antigens from individuals infected with HIV.

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