RESUMO
Legume-Rhizobium spp. symbiosis requires signaling between the symbiotic partners and differential expression of plant genes during nodule development. Previously, we cloned a gene encoding a putative ß-carotene hydroxylase (GmBCH1) from soybean (Glycine max) whose expression increased during nodulation with Bradyrhizobium japonicum. In this work, we extended our study to three GmBCHs to examine their possible role(s) in nodule development, as they were additionally identified as nodule specific, along with the completion of the soybean genome. In situ hybridization revealed the expression of three GmBCHs (GmBCH1, GmBCH2, and GmBCH3) in the infected cells of root nodules, and their enzymatic activities were confirmed by functional assays in Escherichia coli. Localization of GmBCHs by transfecting Arabidopsis (Arabidopsis thaliana) protoplasts with green fluorescent protein fusions and by electron microscopic immunogold detection in soybean nodules indicated that GmBCH2 and GmBCH3 were present in plastids, while GmBCH1 appeared to be cytosolic. RNA interference of the GmBCHs severely impaired nitrogen fixation as well as nodule development. Surprisingly, we failed to detect zeaxanthin, a product of GmBCH, or any other carotenoids in nodules. Therefore, we examined the possibility that most of the carotenoids in nodules are converted or cleaved to other compounds. We detected the expression of some carotenoid cleavage dioxygenases (GmCCDs) in wild-type nodules and also a reduced amount of zeaxanthin in GmCCD8-expressing E. coli, suggesting cleavage of the carotenoid. In view of these findings, we propose that carotenoids such as zeaxanthin synthesized in root nodules are cleaved by GmCCDs, and we discuss the possible roles of the carotenoid cleavage products in nodulation.
Assuntos
Glycine max/fisiologia , Oxigenases de Função Mista/genética , Oxigenases de Função Mista/metabolismo , Proteínas de Plantas/metabolismo , Nódulos Radiculares de Plantas/enzimologia , Arabidopsis/genética , Citosol/enzimologia , Dioxigenases/metabolismo , Escherichia coli/genética , Regulação da Expressão Gênica de Plantas , Fixação de Nitrogênio/genética , Oxirredutases/metabolismo , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Plastídeos/enzimologia , Protoplastos/metabolismo , Interferência de RNA , Nódulos Radiculares de Plantas/genética , Xantofilas/análise , ZeaxantinasRESUMO
Diabetes mellitus (DM) is a progressive, chronic metabolic disorder characterized by high oxidative stress, which can lead to cardiac damage. Methionine sulfoxylation (MetO) of proteins by excessive reactive oxygen species (ROS) can impair the basic functionality of essential cellular proteins, contributing to heart failure. Methionine sulfoxide reductase B2 (MsrB2) can reverse oxidation induced MetO in mitochondrial proteins, so we investigated its role in diabetic cardiomyopathy. We observed that DM-induced heart damage in diabetic mice model is characterized by increased ROS, increased protein MetO with mitochondria structural pathology, and cardiac fibrosis. In addition, MsrB2 was significantly increased in mouse DM cardiomyocytes, supporting the induction of a protective process. Further, MsrB2 directly induces Parkin and LC3 activation (mitophagy markers) in cardiomyocytes. In MsrB2, knockout mice displayed abnormal electrophysiological function, as determined by ECG analysis. Histological analysis confirmed increased cardiac fibrosis and disrupted cardiac tissue in MsrB2 knockout DM mice. We then corroborated our findings in human DM heart samples. Our study demonstrates that increased MsrB2 expression in the heart protects against diabetic cardiomyopathy.
RESUMO
OBJECTIVES: Monkeypox outbreaks in nonendemic countries have been reported since early May 2022. The first case of monkeypox in the Republic of Korea was confirmed in a patient who traveled to Europe in June 2022, and an attempt was made to isolate and identify the monkeypox virus (MPXV) from the patient's specimens. METHODS: Clinical specimens from the patient were inoculated in Vero E6 cells. The isolated virus was identified as MPXV by the observation of cytopathic effects on Vero E6 cells, transmission electron microscopy, conventional polymerase chain reaction (PCR), and sequencing of PCR products. RESULTS: Cytopathic effects were observed in Vero E6 cells that were inoculated with skin lesion swab eluates. After multiple passages from the primary culture, orthopoxvirus morphology was observed using transmission electron microscopy. In addition, both MPXV-specific (F3L and ATI) and orthopoxvirus-specific genes (A39R, B2R, and HA) were confirmed by conventional PCR and Sanger sequencing. CONCLUSION: These results indicate the successful isolation and identification of MPXV from the first patient in the Republic of Korea. The isolated virus was named MPXV-ROK-P1-2022.
RESUMO
OBJECTIVES: Following reports of patients with unexplained pneumonia at the end of December 2019 in Wuhan, China, the causative agent was identified as coronavirus (SARS-CoV-2), and the 2019 novel coronavirus disease was named COVID-19 by the World Health Organization. Putative patients with COVID-19 have been identified in South Korea, and attempts have been made to isolate the pathogen from these patients. METHODS: Upper and lower respiratory tract secretion samples from putative patients with COVID-19 were inoculated onto cells to isolate the virus. Full genome sequencing and electron microscopy were used to identify the virus. RESULTS: The virus replicated in Vero cells and cytopathic effects were observed. Full genome sequencing showed that the virus genome exhibited sequence homology of more than 99.9% with SARS-CoV-2 which was isolated from patients from other countries, for instance China. Sequence homology of SARS-CoV-2 with SARS-CoV, and MERS-CoV was 77.5% and 50%, respectively. Coronavirus-specific morphology was observed by electron microscopy in virus-infected Vero cells. CONCLUSION: SARS-CoV-2 was isolated from putative patients with unexplained pneumonia and intermittent coughing and fever. The isolated virus was named BetaCoV/Korea/KCDC03/2020.
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BACKGROUND: Studies on the results of arthroscopic repair of massive rotator cuff tears have reported widely varied prognoses. Among other factors, the sizable discrepancy can be attributable to the fact that the current definition of massive rotator cuff tears covers an extensive area of tendons. HYPOTHESIS: Functional and radiological results according to subgroups would show significant inter-subgroup differences preoperatively and postoperatively. STUDY DESIGN: Cohort study; Level of evidence, 2. METHODS: A total of 104 patients who required arthroscopic repair for massive rotator cuff tears were prospectively evaluated. The patients were allocated into 3 groups according to tendon involvement as diagnosed by preoperative magnetic resonance imaging: group 1 (anterosuperior type involving the subscapularis and supraspinatus), group 2 (posterosuperior type involving the infraspinatus and supraspinatus), and group 3 (anteroposterior type involving the subscapularis, supraspinatus, and infraspinatus). We compared functional results (at 2 years postoperatively) and radiological findings (at 1 year postoperatively) for each group. RESULTS: There were 34 patients in group 1, 54 in group 2, and 16 in group 3. In all 3 groups, functional results significantly improved after surgery. There were no statistically significant intergroup differences in functional results among the 3 groups. On the radiological evaluations, each group (groups 1, 2, and 3) showed a significantly different result in the preoperative acromiohumeral distance (AHD) (7.19, 5.44, and 5.22 mm, respectively), tear size (38.8, 39.3, and 46.4 mm, respectively), extent of retraction (33.9, 40.0, and 41.4 mm, respectively), postoperative AHD (8.92, 7.37, and 6.71 mm, respectively), and retear rate (23.5%, 51.9%, and 56.2%, respectively) ( P < .001 for all). CONCLUSION: Massive rotator cuff tears can be divided into 3 types: anterosuperior (group 1), posterosuperior (group 2), and anteroposterior (group 3). Each group has distinctive characteristics and shows different results in the preoperative AHD, tear size, extent of retraction, postoperative AHD, and retear rate, which provide a reasonable basis for categorization. So far, massive rotator cuff tears have only been broadly defined, consequently being understood as a single category by many. However, to clearly understand and evaluate this injury, we suggest identifying differences within the category through proper subclassification.
Assuntos
Lesões do Manguito Rotador/classificação , Lesões do Manguito Rotador/etiologia , Idoso , Estudos de Coortes , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Radiografia , Lesões do Manguito Rotador/diagnóstico por imagem , Lesões do Manguito Rotador/fisiopatologiaRESUMO
Eruptive pseudoangiomatosis (EPA) is a rare, self-limiting exanthem, which is known to occur primarily in children. It is characterized by an eruption of distinctive erythematous angioma-like papules often surrounded by a pale halo, with histological findings distinct from that of true angiomas. We describe three women with angioma-like papules. The biopsy specimens from them showed a unique histological appearance consisting of dilated dermal blood vessels with plump endothelial cell and perivascular lymphocytes. On the basis of the clinical and histological findings, we present the adult cases of eruptive pseudoangiomatosis and their etiology.
Assuntos
Angiomatose/diagnóstico , Dermatopatias Vasculares/diagnóstico , Idoso , Angiomatose/patologia , Biópsia , Capilares/patologia , Diagnóstico Diferencial , Células Endoteliais/patologia , Endotélio Vascular/patologia , Exantema/diagnóstico , Feminino , Hemangioma/diagnóstico , Humanos , Linfócitos/patologia , Pessoa de Meia-Idade , Dermatopatias Vasculares/patologia , Neoplasias Cutâneas/diagnósticoRESUMO
Cutaneous calcification may be divided into four major categories: (i) dystrophic; (ii) metastatic; (iii) idiopathic; and (iv) iatrogenic. Dystrophic calcification is the most common type of calcinosis cutis and is associated with a variety of diseases. It most notably occurs in connective tissue diseases. Diffuse and limited cutaneous systemic sclerosis is an example of connective tissue diseases that frequently show calcinosis. We experienced a case of fingertip calcinosis cutis associated with Raynaud's phenomenon. The patient had no previous trauma, skin lesion or systemic connective tissue disease. We propose that calcinosis cutis of the fingertip may result from chronic ischemic injury caused by Raynaud's phenomenon.
Assuntos
Calcinose/complicações , Dedos/patologia , Dermatoses da Mão/complicações , Doença de Raynaud/complicações , Adulto , Calcinose/patologia , Cálcio/análise , Feminino , Seguimentos , Dermatoses da Mão/patologia , Humanos , Ceratose/patologiaRESUMO
Acervuli are calcified concretions in the pineal gland (PG). Particularly interesting are their incidence and size, which are believed to affect neurological disorders and many physiological functions of PG such as regulating circadian rhythm. Despite long investigations for a century, detailed growth mechanism of acervuli has yet to be studied. Here we study the growth morphology of acervuli in human PGs by a direct visualization in 3-dimension (3-D) using a synchrotron X-ray imaging method. For an entire PG, non-aggregated acervuli show Gaussian distribution in size with 47±28 µm. The 3-D volume rendered images of acervuli reveal that the bumpy surfaces developed by lamination result in the mulberry-like structure. In addition, coalescence of multiple acervuli leads to large-scale lamination on the whole aggregate. We suggest a novel hypothesis on the growth patterns of acervuli by their nucleation density (N(d)): i) mulberry-like structure at low N(d), and ii) large-scale lamination on an aggregate at high N(d).
Assuntos
Extensões da Superfície Celular/ultraestrutura , Glândula Pineal/crescimento & desenvolvimento , Glândula Pineal/ultraestrutura , Humanos , Modelos AnatômicosRESUMO
Disrupted cortical cytoarchitecture in cerebellum is a typical pathology in reeler. Particularly interesting are structural problems at the cellular level: dendritic morphology has important functional implication in signal processing. Here we describe a combinatorial imaging method of synchrotron X-ray microtomography with Golgi staining, which can deliver 3-dimensional(3-D) micro-architectures of Purkinje cell(PC) dendrites, and give access to quantitative information in 3-D geometry. In reeler, we visualized in 3-D geometry the shape alterations of planar PC dendrites (i.e., abnormal 3-D arborization). Despite these alterations, the 3-D quantitative analysis of the branching patterns showed no significant changes of the 77 ± 8° branch angle, whereas the branch segment length strongly increased with large fluctuations, comparing to control. The 3-D fractal dimension of the PCs decreased from 1.723 to 1.254, indicating a significant reduction of dendritic complexity. This study provides insights into etiologies and further potential treatment options for lissencephaly and various neurodevelopmental disorders.
Assuntos
Córtex Cerebral/patologia , Células de Purkinje/patologia , Animais , Córtex Cerebral/diagnóstico por imagem , Córtex Cerebral/crescimento & desenvolvimento , Dendritos/diagnóstico por imagem , Dendritos/patologia , Modelos Animais de Doenças , Fractais , Imageamento Tridimensional , Lisencefalia/patologia , Malformações do Desenvolvimento Cortical/patologia , Camundongos , Camundongos Mutantes Neurológicos , Células de Purkinje/diagnóstico por imagem , Microtomografia por Raio-XRESUMO
Ultrastructural deformities were monitored after cryopreservation of F1 (B6CBAF1) mouse ovaries by either slow freezing or vitrification. Vacuole formation in the ooplasm, zona pellucida, and the cytoplasm of follicular cells, and mitochondrial deformity were detected. These types of cryodamage demonstrated protocol specificity.
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Criopreservação/métodos , Congelamento/efeitos adversos , Folículo Ovariano/ultraestrutura , Ovário , Animais , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , Folículo Ovariano/patologia , Zona Pelúcida/patologia , Zona Pelúcida/ultraestruturaRESUMO
Viral wart infections constitute one of the most common pediatric skin diseases, and various modalities have been used to manage them. Although pulsed dye laser therapy is known to be a safe and efficacious modality, the reported cure rates for this method have varied, and no studies have reported treatment of pediatric patients alone. This prospective, nonblinded, nonrandomized study was performed to evaluate the efficacy and safety of pulsed dye laser therapy for pediatric warts. We found that this method is safe, relatively effective, and worth considering as an additional therapeutic option for viral warts in children, although not as a first-line therapy.