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1.
Chinese Journal of Biotechnology ; (12): 2334-2358, 2023.
Article in Chinese | WPRIM | ID: wpr-981205

ABSTRACT

As a generally-recognized-as-safe microorganism, Saccharomyces cerevisiae is a widely studied chassis cell for the production of high-value or bulk chemicals in the field of synthetic biology. In recent years, a large number of synthesis pathways of chemicals have been established and optimized in S. cerevisiae by various metabolic engineering strategies, and the production of some chemicals have shown the potential of commercialization. As a eukaryote, S. cerevisiae has a complete inner membrane system and complex organelle compartments, and these compartments generally have higher concentrations of the precursor substrates (such as acetyl-CoA in mitochondria), or have sufficient enzymes, cofactors and energy which are required for the synthesis of some chemicals. These features may provide a more suitable physical and chemical environment for the biosynthesis of the targeted chemicals. However, the structural features of different organelles hinder the synthesis of specific chemicals. In order to ameliorate the efficiency of product biosynthesis, researchers have carried out a number of targeted modifications to the organelles grounded on an in-depth analysis of the characteristics of different organelles and the suitability of the production of target chemicals biosynthesis pathway to the organelles. In this review, the reconstruction and optimization of the biosynthesis pathways for production of chemicals by organelle mitochondria, peroxisome, golgi apparatus, endoplasmic reticulum, lipid droplets and vacuole compartmentalization in S. cerevisiae are reviewed in-depth. Current difficulties, challenges and future perspectives are highlighted.


Subject(s)
Saccharomyces cerevisiae/metabolism , Saccharomyces cerevisiae Proteins/metabolism , Golgi Apparatus/metabolism , Metabolic Engineering , Vacuoles/metabolism
2.
J. venom. anim. toxins incl. trop. dis ; 27: e20200106, 2021. tab, graf, ilus
Article in English | LILACS, VETINDEX | ID: biblio-1154774

ABSTRACT

Avian pathogenic Escherichia coli (APEC) isolated from avian cellulitis lesions produces a toxin, named Escherichia coli vacuolating factor (ECVF), that causes cell vacuolization and induces inflammatory response in broiler chicken. Methods We investigated the intracellular activities of ECVF in avian fibroblasts using fluorescence staining, electron microscopy, MTT and LDH measurements. As ECVF act specifically in avian cells, we performed blotting assay followed by mass spectrometry to better understand its initial intracellular protein recognition. Results ECVF induced actin contraction, mitochondrial damage and membrane permeability alterations. Ultrastructural analysis showed intracellular alterations, as nuclear lobulation and the presence of degraded structures inside the vacuoles. Moreover, ECVF induced cell death in fibroblasts. ECVF-biotin associates to at least two proteins only in avian cell lysates: alpha-actinin 4 and vinculin, both involved in cytoskeleton structure. Conclusion These findings demonstrated that ECVF plays an important role in avian cellulitis, markedly in initial steps of infection. Taken together, the results place this toxin as a target for drug and/or vaccine development, instead of the use of large amounts antibiotics.(AU)


Subject(s)
Animals , Vacuoles , Actin Cytoskeleton , Chickens , Actins , Escherichia coli , Fibroblasts , Cellulitis
3.
Rev. med. vet. zoot ; 67(2): 136-148, May-Aug. 2020. tab, graf
Article in Spanish | LILACS, COLNAL | ID: biblio-1180950

ABSTRACT

RESUMEN Las enfermedades gastrointestinales equinas tienen una alta incidencia con un pronóstico variable en la práctica clínica. La mayoría de los estudios se limitan a describir lesiones ulcerativas y lesiones inflamatorias. El objetivo de este estudio fue evaluar el potencial diagnóstico complementario de la cromoendoscopia convencional en la mucosa gas-troesofágica y duodenal proximal del equino. El estudio incluyó 20 caballos criollos colombianos de ambos sexos (12 hembras y 8 machos), con edades entre 5 y 20 años, peso entre 250 y 350 kilogramos, condición corporal 4-5/9 y con historial de alteraciones digestivas en los últimos 3 meses; quienes previo a la evaluación por gastroscopia y cromoendoscopia se sometieron a ayuno (sólidos 12h y líquidos 4h) y sedación (xilacina 0,5 mg/kg/iv). Se utilizaron tinciones como rojo fenol, lugol, índigo carmín, azul de metileno y ácido acético y se tomaron biopsias de los segmentos que mostraron reacción. El azul de metileno reveló 52% de las lesiones, el lugol 19%; por su parte, el rojo fenol, el índigo carmín y el ácido acético revelaron el 9,5% restante. El epitelio escamoso fue el más afectado (66,6%), el glandular (19%), antro pilórico (9,5%) y duodeno proximal (4,7%). Los hallazgos histopatológicos fueron hiperplasia, hipertrofia, hiperqueratosis, congestión, degeneración vacuolar, infiltrados celulares, fibrosis, necrosis y atrofia en diferentes grados de severidad. La cromoendoscopia reveló lesiones prematuras, que pasaron desapercibidas con las técnicas convencionales de endoscopia del tracto digestivo. Este es el primer estudio que emplea la cromoendoscopia en equinos; a pesar de que la técnica mejoró la visualización y facilitó la ubicación y descripción de lesiones ulcerativas prematuras a través de la histopatología, se recomiendan mayores estudios controlados y con un número más amplio de muestras.


ABSTRACT Equine gastrointestinal diseases have a high occurrence with a variable prognostic in clinic practice. Most of the studies limits to describe ulcerative and inflammatory lesions. The objective of this study was to evaluate the potential complementary diagnostic of conventional chromoendoscopy on the gastroesophageal and proximal duodenal mucosa of the equine. 20 Colombian creole horses, of both sexes (12 females and 8 males), with ages between 5 and 20 years old, weight between 250 and 350 kilograms, body condition 4-5/9, that had presented digestive alterations in the last 3 months, were subjected to fasting (solids 12h and liquids 4h) and sedated (xylazine 0,5 mg/kg/iv) to be evaluated by gastroscopy and chromoendoscopy, using for stains phenol red, lugol, indigo carmine, methylene blue and acetic acid, taking biopsy samples of the segments that showed reaction. The methylene blue revealed 52%, lugol 19%, and phenol red, indigo carmine and acetic acid revealed only 9,5% of the lesions, being the squamous epithelium the most affected (66,6%), glandular epithelium (19%), pyloric antrum (9,5%) and proximal duodenum (4,7%), where histopathological findings were hyperplasia, hypertrophy, hyperkeratosis, congestion, vacuolar degeneration, cellular infiltrates, fibrosis, necrosis and atrophy in different degrees of severity. Chromoendoscopy revealed lesions premature, which go unnoticed with conventional light endoscopy techniques. This is the first study using chromoscopy in horses to show that the reagents used allow a better visualization of injuries than the conventional technique, helping histopathological studies and molecular biology to understand ulcerative premature injuries and possible pathophysiological pathways. However, larger controlled studies and a larger number of samples are needed.


Subject(s)
Animals , Wounds and Injuries , Gastroscopy , Coloring Agents , Endoscopy , Horses , Atrophy , Vacuoles , Biopsy , Cells , Carmine , Fasting , Acetic Acid , Phenol , Duodenum , Epithelium , Age and Sex Distribution , Esophagogastric Junction , Hyperplasia , Hypertrophy , Methylene Blue , Mucous Membrane , Necrosis
4.
Arq. bras. oftalmol ; 83(2): 146-148, Mar.-Apr. 2020. graf
Article in English | LILACS | ID: biblio-1088972

ABSTRACT

ABSTRACT Lisch corneal dystrophy is a rare corneal disease characterized by the distinctive feature of highly vacuolated cells. Although this feature is important, the nature of these vacuoles within corneal cells remains unknown. Here, we sought to analyze corneal cells from a patient diagnosed with Lisch dystrophy to characterize the vacuoles within these cells. Analyses using histopathology examination, confocal microscopy, and transmission electron microscopy were all consistent with previous descriptions of Lisch cells. Importantly, the vacuoles within these cells appeared to be autophagosomes and autolysosomes, and could be stained with an anti-microtubule-associated protein 1A/1B-light chain 3 (LC3) antibody. Taken together, these findings indicate that the vacuoles we observed within superficial corneal cells of a patient with Lisch corneal dystrophy constituted autophagosomes and autolysosomes; this finding has not been previously reported and suggests a need for further analyses to define the role of autophagy in this ocular disease.


RESUMO A distrofia corneana de Lisch é uma doença rara, caracterizada principalmente pela presença de células altamente vacuoladas. Embora esta característica seja importante, a natureza desses vacúolos dentro das células da córnea permanece des conhecida. Aqui, procuramos analisar as células da córnea de um paciente diagnosticado com distrofia de Lisch para caracte rizar os vacúolos dentro dessas células. Análises utilizando exame histopatológico, microscopia confocal e microscopia eletrônica de transmissão foram todas consistentes com descrições previas de células de Lisch. Importante, os vacúolos dentro dessas células pareciam ser autofagossomos e autolisossomos, e po deriam ser corados com um anticorpo proteico 1A/1B-cadeia leve 3 (LC3) da proteína anti-microtúbulo associado a microtúbulos. Em conjunto, esses achados indicam que os vacúolos observados nas células superficiais da córnea de um paciente com distrofia corneana de Lisch constituíram autofagossomos e autolisossomos. Esse achado não foi relatado anteriormente e sugere a necessidade de mais análises para definir o papel da autofagia nessa doença ocular.


Subject(s)
Humans , Female , Adult , Vacuoles/pathology , Corneal Dystrophies, Hereditary/pathology , Autophagosomes/pathology , Corneal Dystrophies, Hereditary/diagnostic imaging , Microscopy, Confocal/methods , Corneal Opacity/pathology , Corneal Opacity/diagnostic imaging , Tomography, Optical Coherence/methods , Microscopy, Electron, Transmission/methods , Microautophagy
5.
Yonsei Medical Journal ; : 262-266, 2020.
Article in English | WPRIM | ID: wpr-811468

ABSTRACT

The World Health Organization 2016 edition assigned anaplastic lymphoma kinase (ALK) rearrangement-associated renal cell carcinoma (ALK-RCC) as an emerging renal tumor entity. Identifying ALK-RCC is important because ALK inhibitors have been shown to be effective in treatment. Here, we report the case of a 14-year-old young man with ALK-RCC. Computed tomography revealed a well-demarcated 5.3-cm enhancing mass at the upper pole of the left kidney. There was no further history or symptoms of the sickle-cell trait. The patient underwent left radical nephrectomy. Pathologically, the mass was diagnosed as an unclassified RCC. Targeted next-generation sequencing identified a TPM3-ALK fusion gene. The present report and literature review demonstrate that TPM3-ALK RCC may be associated with distinct clinicopathological features. Microscopically, the tumors showed diffuse growth and tubulocystic changes with inflammatory cell infiltration. Tumor cells were dis-cohesive and epithelioid with abundant eosinophilic cytoplasm and cytoplasmic vacuoles. If morphological features and TFE3 expression are present in adolescent and young patients, molecular tests for ALK translocation should be performed. This awareness is critically important, because ALK rearrangement confers sensitivity to ALK inhibitors.


Subject(s)
Adolescent , Humans , Carcinoma, Renal Cell , Cytoplasm , Eosinophils , Gene Rearrangement , Kidney , Lymphoma , Nephrectomy , Phosphotransferases , Vacuoles , World Health Organization
6.
Biomédica (Bogotá) ; 39(supl.2): 26-31, ago. 2019. graf
Article in Spanish | LILACS | ID: biblio-1038825

ABSTRACT

Resumen Los pacientes con lepra lepromatosa que han recibido tratamiento durante años, usualmente requieren seguimiento con biopsias de piel para detectar lesiones persistentes o si la baciloscopia es positiva, incluso si los valores son menores que los iniciales. Se presenta el caso de una mujer de 48 años de edad con lepra lepromatosa de 15 años de evolución, índice bacilar de 4 en el extendido directo y en la biopsia, que recibió tratamiento con múltiples medicamentos durante 32 meses, aunque lo recomendado por la Organización Mundial de la Salud (OMS) es una duración de 12 meses. Se tomó una biopsia de piel para determinar si la enfermedad estaba activa. Se observó inflamación dérmica difusa con numerosas células gigantes de tipo cuerpo extraño y macrófagos vacuolados (células de Virchow). Estas células, CD68 positivas, contenían material granular ácido-alcohol resistente positivo con inmunohistoquímica para BCG. Se encontraron bacilos fragmentados y el índice bacilar fue de 2. Se interpretó como una forma residual de lepra lepromatosa y se concluyó que la paciente no requería prolongar el tratamiento con múltiples medicamentos. Este perfil histológico se ha observado en casos similares, pero sin datos clínicos estas biopsias representan un reto diagnóstico. La acumulación de lípidos en estas células gigantes se debe a la destrucción bacilar y a la fusión de macrófagos vacuolados. Se revisó el papel de los lípidos del bacilo y del huésped en la patogenia de la lepra lepromatosa. En estos casos, no es necesario extender los 12 meses de tratamiento con múltiples medicamentos recomendados por la OMS. En el seguimiento de los pacientes, se recomienda contar con los hallazgos clínicos, la baciloscopia, la biopsia anual de piel y los títulos IgM antiglucolípido fenólico.


Abstract Patients with lepromatous leprosy that have received treatment for many years usually get follow up biopsies for persistent skin lesions or positive bacilloscopy even if the values are lower than in the initial bacilloscopy. We report the case of a 48-year old woman with long-standing lepromatous leprosy of 15 years of evolution, with a bacterial index of 4 in the direct smear and the initial skin biopsy. The patient was treated with multidrug therapy for 32 months although the treatment recommended by the World Health Organization (WHO) is only for 12 months. A skin biopsy was taken to determine if there was an active disease. We observed a diffuse dermal inflammation with numerous foreign body giant cells and vacuolated macrophages (Virchow´s cells). These cells contained granular acid-fast material that was also positive with immunohistochemistry for BCG. There were fragmented bacilli and the BI was 2. These cells were also strongly positive for CD68. The biopsy was interpreted as a residual form of lepromatous leprosy that did not require further multidrug therapy. We have observed similar histological profiles in several cases. The lack of clinical data makes it a histological challenge. The accumulation of lipids in these giant cells is due to bacillary destruction and fusion of vacuolated macrophages. We discuss here the role of bacillary and host lipids in the pathogenesis of lepromatous leprosy. We concluded that there was no need to extend the 12-month multidrug therapy recommended by WHO. Clinical findings, bacilloscopy, annual skin biopsy, and anti-phenolic glycolipid-I IgM titers are recommended procedures for the follow-up of these patients.


Subject(s)
Female , Humans , Middle Aged , Skin/pathology , Leprosy, Lepromatous/pathology , Giant Cells, Foreign-Body/pathology , Foam Cells/pathology , Skin/microbiology , Vacuoles , Biopsy , Antigens, Differentiation, Myelomonocytic/analysis , Leprosy, Lepromatous/drug therapy , Antigens, CD/analysis , Giant Cells, Foreign-Body/microbiology , Giant Cells, Foreign-Body/chemistry , Cell Wall/chemistry , Drug Therapy, Combination , Host-Pathogen Interactions , Foam Cells/microbiology , Foam Cells/chemistry , Leprostatic Agents/therapeutic use , Lipids/analysis , Mycobacterium leprae/isolation & purification , Mycobacterium leprae/chemistry
7.
Article in English | WPRIM | ID: wpr-741205

ABSTRACT

Secretory carcinoma of the salivary gland (SC) is a newly introduced rare salivary gland tumor that shares histological, immunohistochemical, and genetic characteristics with secretory carcinoma of the breast. Here, we report the cytologic features of two cases of SC confirmed by surgical resection. In these two cases, SC was incidentally detected in a 64-year-old female and a 56-yearold male. Fine needle aspiration cytology revealed nests of tumor cells with a papillary or glandular structure floating in mucinous secretions. The tumor cells demonstrated uniform, round, smooth nuclear contours and distinct nucleoli. Multiple characteristic cytoplasmic vacuoles were revealed. Singly scattered tumor cells frequently showed variable sized cytoplasmic vacuoles. The cytopathologic diagnosis of SC should be considered when characteristic cytological findings are revealed. Further immunohistochemistry and gene analyses are helpful to diagnose SC.


Subject(s)
Female , Humans , Male , Middle Aged , Biopsy, Fine-Needle , Breast , Clothing , Cytoplasm , Diagnosis , Immunohistochemistry , Mammary Analogue Secretory Carcinoma , Mucins , Salivary Glands , Vacuoles
8.
Article in English | WPRIM | ID: wpr-764348

ABSTRACT

BACKGROUND AND PURPOSE: GNE myopathy is a rare progressive myopathy caused by biallelic mutations in the GNE gene, and frequently accompanied by rimmed vacuoles in muscle pathology. The initial symptom of foot drop or hip-girdle weakness eventually spreads to all limbs over a period of decades. Recent advances in pathophysiologic research have facilitated therapeutic trials aimed at resolving the core biochemical defect. However, there remains unsettled heterogeneity in its natural course, which confounds the analysis of therapeutic outcomes. We performed the first large-scale study of Korean patients with GNE myopathy. METHODS: We gathered the genetic and clinical profiles of 44 Korean patients with genetically confirmed GNE myopathy. The clinical progression was estimated retrospectively based on a patient-reported questionnaire on the status of the functional joint sets and daily activities. RESULTS: The wrist and neck were the last joints to lose antigravity functionality irrespective of whether the weakness started from the ankle or hip. Two-thirds of the patients could walk either independently or with an aid. The order of losing daily activities could be sorted from standing to eating. Patients with limb-girdle phenotype showed an earlier age at onset than those with foot-drop onset. Patients with biallelic kinase domain mutations tended to progress more rapidly than those with epimerase and kinase domain mutations. CONCLUSIONS: The reported data can guide the clinical management of GNE myopathy, as well as provide perspective to help the development of clinical trials.


Subject(s)
Humans , Age of Onset , Ankle , Disease Progression , Eating , Extremities , Foot , Hip , Joints , Muscular Diseases , Muscular Dystrophies, Limb-Girdle , Neck , Pathology , Phenotype , Phosphotransferases , Population Characteristics , Retrospective Studies , Surveys and Questionnaires , Vacuoles , Wrist
9.
Chinese Journal of Biotechnology ; (12): 1424-1432, 2019.
Article in Chinese | WPRIM | ID: wpr-771787

ABSTRACT

Important progress has been made in the interpretation of subcellular location, ion transport characteristics and biological functions of endosomal Na⁺,K⁺/H⁺ antiporter in Arabidopsis thaliana. The endosomal Na⁺,K⁺/H⁺ antiporter contain two members, AtNHX5 and AtNHX6, whose amino acid sequence similarity is 78.7%. Studies have shown that AtNHX5 and AtNHX6 are functionally redundant, and they are all located in Golgi, trans-Golgi network (TGN), endoplasmic reticulum (ER) and prevacuolar compartment (PVC). AtNHX5 and AtNHX6 are critical for salt tolerance stress and the homeostasis of pH and K⁺. It has been reported that there are conservative acidic amino acid residues that can regulate their ion activity in the endosomal NHXs transmembrane domain, which plays a decisive role in their own functions. The results of the latest research indicate that endosomal NHXs affect vacuolar transport and protein storage, and participate in the growth of auxin-mediated development in A. thaliana. In this paper, the progress of subcellular localization, ion transport, function and application of endosomal NHXs in A. thaliana was summarized.


Subject(s)
Arabidopsis , Arabidopsis Proteins , Endosomes , Sodium-Hydrogen Exchangers , Vacuoles
10.
Biomédica (Bogotá) ; 38(supl.2): 135-143, ago. 2018. tab, graf
Article in English | LILACS | ID: biblio-1038798

ABSTRACT

ABSTRACT Introduction: Dengue virus replication has been considered mainly cytoplasmic, however, studies indicate that some flaviviruses may use the intranuclear pathway as part of the machinery that the virus uses to increase infection capacity in the host cell. This paper describes alterations at nuclear level in the cell infected with dengue, which are likely involved in the virus replication processes. Objective: This paper addresses the ultrastructural observations of C6/36 cells of the Aedes albopictus mosquito infected with dengue virus type 2. Materials and methods: C6/36 cells were infected in culture medium with the serum of a patient positively diagnosed for dengue 2. Subsequently, the cells were incubated for 10 days and the cytopathic effect was assessed. The cells were processed for immunofluorescence assays and transmission electron microscopy. Results: The immunofluorescence assays confirmed the presence of viral protein E associated with cellular syncytia in the culture. In the ultrastructural study, the infected cells showed vesicular-tubular structures and dilated cisterns of the endoplasmic reticulum at the cytoplasmic level. Viral particles were found exclusively in cytoplasm localized within the vacuoles. Nuclei of cellular syncytia showed membrane structures arranged in a circular shape and, in some cases, these syncytia displayed lysis; in no case viral particles were observed at the nuclear level. Conclusions: The ultrastructural alterations of nuclei in cells infected with the dengue virus using electron microscopy techniques had not been reported before, as far as we know. It is likely that such modifications are associated with replicative processes at an intranuclear level as an alternate replication mechanism.


RESUMEN Introducción. La replicación del virus del dengue se ha considerado principalmente citoplásmica; sin embargo, en diversos estudios se ha informado que algunos flavivirus pueden utilizar factores intranucleares como parte de la maquinaria que utilizan para aumentar la capacidad de infección en la célula huésped. En este trabajo se describen las alteraciones a nivel nuclear en células infectadas con dengue, probablemente involucradas en procesos de replicación viral. Objetivo. Presentar las observaciones ultraestructurales de células C6/36 de Aedes albopictus infectadas con el virus del dengue de tipo 2. Materiales y métodos. Se infectaron células C6/36 con suero de un paciente con diagnóstico de dengue 2; posteriormente, se mantuvieron en medio de cultivo durante 10 días y se evaluó el efecto citopático. Las células se procesaron para los ensayos de inmunofluorescencia y microscopía electrónica de transmisión, con el fin de hacer el estudio ultraestructural. Resultados. Los ensayos de inmunofluorescencia confirmaron la presencia de la proteína E viral asociada con sincitios celulares en el cultivo. En el estudio ultraestructural, las células infectadas tenían estructuras vesiculares y tubulares, y cisternas dilatadas del retículo endoplásmico en el citoplasma. Las partículas virales se encontraron exclusivamente en vacuolas localizadas en el citoplasma. Los núcleos de los sincitios celulares contenían estructuras de membrana dispuestas en forma circular y, en algunos casos, dichos sincitios presentaban lisis. En ningún caso se observaron partículas virales en el núcleo. Conclusiones. No se habían reportado alteraciones ultraestructurales en los núcleos de células infectadas con el virus del dengue detectadas mediante técnicas de microscopia electrónica. Es probable que tales modificaciones estén asociadas con procesos intranucleares de replicación como un mecanismo alternativo.


Subject(s)
Animals , Humans , Cell Nucleus/ultrastructure , Cytopathogenic Effect, Viral , Dengue Virus/physiology , Vacuoles/virology , Viremia/virology , Virus Replication , Microscopy, Electron , Giant Cells/virology , Cell Line , Viral Envelope Proteins/analysis , Aedes/cytology , Cytoplasm/virology , Dengue/virology , Dengue Virus/isolation & purification , Microscopy, Fluorescence
11.
Article in English | WPRIM | ID: wpr-727861

ABSTRACT

Extra-hypothalamic growth hormone-releasing hormone (GHRH) plays an important role in reproduction. To study the treatment effect of Grin (a novel hGHRH homodimer), the infertility models of 85 male Chinese hamsters were established by intraperitoneally injecting 20 mg/kg of cyclophosphamide once in a week for 5 weeks and the treatment with Grin or human menopausal gonadotropin (hMG) as positive control was evaluated by performing a 3-week mating experiment. 2–8 mg/kg of Grin and 200 U/kg of hMG showed similar effect and different pathological characteristics. Compared to the single cyclophosphamide group (0%), the pregnancy rates (H-, M-, L-Grin 26.7, 30.8, 31.3%, and hMG 31.3%) showed significant difference, but there was no difference between the hMG and Grin groups. The single cyclophosphamide group presented loose tubules with pathologic vacuoles and significant TUNEL positive cells. Grin induced less weight of body or testis, compactly aligned tubules with little intra-lumens, whereas hMG caused more weight of body or testis, enlarging tubules with annular clearance. Grin presented a dose-dependent manner or cell differentiation-dependentincrease in testicular GHRH receptor, and did not impact the levels of blood and testicular GH, testosterone. Grin promotes fertility by proliferating and differentiating primitive cells through up-regulating testicular GHRH receptor without triggering GH secretion, which might solve the etiology of oligoasthenozoospermia.


Subject(s)
Animals , Cricetinae , Humans , Male , Male , Cricetulus , Cyclophosphamide , Fertility , Gonadotropins , Growth Hormone-Releasing Hormone , In Situ Nick-End Labeling , Infertility , Infertility, Male , Pregnancy Rate , Reproduction , Testis , Testosterone , Vacuoles
12.
Yonsei Medical Journal ; : 698-701, 2018.
Article in English | WPRIM | ID: wpr-715891

ABSTRACT

Limb-girdle muscular dystrophies (LGMD) are heterogeneous disorders with autosomal inheritance. Autosomal dominant LGMD mapped to 7q36.3 has been classified as LGMD type 1D (LGMD1D) in the Human Gene Nomenclature Committee Database. LGMD1D is characterized predominantly by limb-girdle weakness and may also show a bulbar symptom in some cases. In the past, the frequency of this disease was uncommon, and this disorder was mainly found in Europe and the United States. However, recently, this disorder has been reported in Asia, including Japan, Korea, and Taiwan. Here, we report on three LGMD1D patients, including one with a novel mutation in DNAJB6, c.298T>A. While two patients complained of limb-girdle weakness, as would be expected, one patient had distal weakness. They had various serum creatine kinase levels. Radiologic findings in one patient showed fatty degeneration and atrophy in the posterior part of distal muscles. Pathologic findings in one of the patients showed rimmed vacuoles. Although LGMD1D is still uncommon in Korea, we discovered three Korean patients with LGMD1D, including one novel mutation in DNAJB6, p.Phe100Ile (c.298T>A).


Subject(s)
Humans , Asia , Atrophy , Creatine Kinase , Europe , Japan , Korea , Muscles , Muscular Diseases , Muscular Dystrophies, Limb-Girdle , Taiwan , United States , Vacuoles , Wills
13.
Journal of Rhinology ; : 118-122, 2018.
Article in Korean | WPRIM | ID: wpr-718262

ABSTRACT

Epithelioid hemangioendothelioma is a rare vascular tumor with intermediate malignity and metastasis risk. It presents epithelioid cells with intracytoplasmic vacuoles and low mitotic activity. Its vascular nature can be confirmed by immunohistochemical studies (vimentin, CD31, CD34, and factor VIII). It is extremely rare in the nasal cavity, with only one case reported on the middle turbinate in Korea. The authors present a case of epithelioid hemangioendothelioma on the choana with a size of 2mm, which easily coult have been misdiagnosed as a blood clot.


Subject(s)
Epistaxis , Epithelioid Cells , Hemangioendothelioma, Epithelioid , Korea , Nasal Cavity , Neoplasm Metastasis , Turbinates , Vacuoles
14.
Mycobiology ; : 114-121, 2018.
Article in English | WPRIM | ID: wpr-729789

ABSTRACT

Mon1 is a guanine nucleotide exchange factor subunit that activates the Ypt7 Rab GTPase and is essential for vacuole trafficking and autophagy in eukaryotic organisms. Here, we identified and characterized the function of Mon1, an ortholog of Saccharomyces cerevisiae Mon1, in a human fungal pathogen, Cryptococcus neoformans. Mutation in mon1 resulted in hypersensitivity to thermal stress. The mon1 deletion mutant exhibited increased sensitivity to cell wall and endoplasmic reticulum stress. However, the mon1 deletion mutant showed more resistance to the antifungal agent fluconazole. In vivo studies demonstrated that compared to the wild-type strain, the mon1 deletion mutant attenuated virulence in the Galleria mellonella insect model. Moreover, the mon1 deletion mutant was avirulent in the murine inhalation model. These results demonstrate that Mon1 plays a crucial role in stress survival and pathogenicity in C. neoformans.


Subject(s)
Humans , Autophagy , Cell Wall , Cryptococcus neoformans , Cryptococcus , Endoplasmic Reticulum Stress , Fluconazole , GTP Phosphohydrolases , Guanine Nucleotide Exchange Factors , Hypersensitivity , Inhalation , Insecta , Saccharomyces cerevisiae , Vacuoles , Virulence
15.
Article in English | WPRIM | ID: wpr-742290

ABSTRACT

This study was carried out to determine the pathogen-causing diarrhoea in sheep Ovis aries in the Qinghai Tibetan Plateau Area, China. A trophozoite was identified as species of ciliate alveolates infecting the sheep based on morphological characteristics examined by microscope. It was mostly spherical, colourless and transparent, with many vesicles. Macronucleus and contractile vacuoles could not be distinguished. Size of the trophozoite was 80–180×70–150 μm and its surface was covered with cilia. Molecular analysis based on sequences of 18S rRNA and ITS genes confirmed the ciliate species as Balantidium coli. According to the literature, there have been many epidemiological investigations of B. coli infection in pigs, monkeys and humans. To our knowledge, this was the first report of B. coli infections in sheep in the Qinghai Tibetan Plateau Area of China, or eleswhere around the world. Importantly, the sheep case was rare but raised our concern that B. coli may spread across species and expand its host range.


Subject(s)
Humans , Balantidium , China , Cilia , Haplorhini , Host Specificity , Macronucleus , Sheep , Sheep, Domestic , Swine , Trophozoites , Vacuoles
16.
Article in English | WPRIM | ID: wpr-728580

ABSTRACT

This study aimed to compare the cellular toxicities of three clinically used dry eye treatments; 3% diquafosol tetrasodium and hyaluronic acid at 0.3 and 0.18%. A methyl thiazolyltetrazoiun (MTT)-based calorimetric assay was used to assess cellular proliferation and a lactate dehydrogenase (LDH) leakage assay to assess cytotoxicity, using Human corneal epithelial cells (HCECs) exposed to 3% diquafosol tetrasodium, 0.3% hyaluronic acid (HA), or 0.18% HA or 1, 6 or 24 h. Cellular morphology was evaluated by inverted phase-contrast light microscopy and electron microscopy, and wound widths were measured 24 h after confluent HCECs were scratched. Diquafosol had a significant, time-dependent, inhibitory effect on HCEC proliferation and cytotoxicity. HCECs treated with diquafosol detached more from the bottoms of dishes and damaged cells showed degenerative changes, such as, reduced numbers of microvilli, vacuole formation, and chromatin of the nuclear remnant condensed along the nuclear periphery. All significantly stimulated reepithelialization of HCECs scratched, which were less observed in diquafosol. Therefore, epithelial toxicity should be considered after long-term usage of diquafosol and in overdose cases, especially in dry eye patients with pre-existing punctated epithelial erosion.


Subject(s)
Humans , Cell Proliferation , Chromatin , Dry Eye Syndromes , Epithelial Cells , Hyaluronic Acid , L-Lactate Dehydrogenase , Microscopy , Microscopy, Electron , Microvilli , Vacuoles , Wound Healing , Wounds and Injuries
17.
Article in English | WPRIM | ID: wpr-649872

ABSTRACT

Human dermal fibroblast is essential in wound healing of the skin through the synthesis of extracellular matrix proteins. With respect to oxidative stress, the effects of remifentanil on human dermal fibroblast have received little attention. Therefore, we investigated the effects of remifentanil on the apoptosis and autophagic reaction of human dermal fibroblasts under oxidative stress. The subjects were divided into the following groups: Control group: cells were incubated at 37℃ in a humidified atmosphere with 5% CO₂. Hydrogen peroxide (H₂O₂) group: cells were exposed to H₂O₂ for 2 h. RPC/H₂O₂ group: cells were pretreated with remifentanil for 2 h and exposed H₂O₂ for 2 h. 3-MA/RPC/H₂O₂ group: cells were pretreated with 3-methyladenine (3-MA) and remifentanil for 1 h and 2 h, respectively. We measured cell viability using MTT assay. Western blot analysis was used to determine the expression levels of proteins associated with apoptosis and autophagy. Quantification of apoptotic cells was performed using flow cytometer analysis, and autophagic vacuoles were observed under a fluorescence microscope. Remifentanil treatment increased the proliferation of human dermal fibroblast and decreased apoptotic cell death, enhancing autophagic activity under oxidative stress. However, 3-MA, the autophagy pathway inhibitor, inhibited the protective effect of remifentanil in oxidative stress. This study demonstrates that remifentanil activated autophagy and decreased apoptotic death of human dermal fibroblasts under oxidative stress. Our results suggest that remifentanil may help in the treatment of oxidative stress.


Subject(s)
Humans , Apoptosis , Atmosphere , Autophagy , Blotting, Western , Cell Death , Cell Survival , Extracellular Matrix Proteins , Fibroblasts , Fluorescence , Hydrogen Peroxide , Oxidative Stress , Skin , Vacuoles , Wound Healing
18.
Anatomy & Cell Biology ; : 187-199, 2017.
Article in English | WPRIM | ID: wpr-50233

ABSTRACT

Tobacco smoking has been identified as an important factor in premature skin aging to detect the histological changes occurred in adult male guinea pig thin skin under the influence of low and high doses of nicotine; which constitutes approximately 0.6%–3.0% of the dry weight of tobacco. Fifteen adult male pigmented guinea pigs were equally divided into three groups: group I, control; group IIA, low dose nicotine treated; 3 mg/kg subcutaneously for 4 weeks; and group IIB, high dose nicotine treated; 6 mg/kg subcutaneously for 4 weeks. Specimens from the back thin skin were processed for light and electron microscopy. Nicotine administration revealed flattened dermo-epidermal junction and reduced rete ridges formation. Collagen bundles were disorganized with increased spaces between them. A reduction in the amount of elastic fibers in the dermis were also observed compared to group I. Ultrastructurally, keratinocytes had hyperchromatic nuclei, intracytoplasmic vacuoles, disruption of desmosomal junctions, irregular tonofilaments distribution, and increased inter-cellular spaces. These changes were more pronounced with high dose nicotine administration. The epidermal thickness was reduced in low dose nicotine administration. But, high dose nicotine administration revealed increased epidermal thickness compared to the control group. Nicotine induced structural changes of adult male guinea pig thin skin. These changes were more pronounced with high dose nicotine administration.


Subject(s)
Adult , Animals , Humans , Male , Collagen , Dermis , Elastic Tissue , Guinea Pigs , Guinea , Intermediate Filaments , Keratinocytes , Microscopy, Electron , Nicotine , Skin Aging , Skin , Smoking , Nicotiana , Vacuoles
19.
Article in English | WPRIM | ID: wpr-110371

ABSTRACT

Thymic adenocarcinoma is extremely rare. Although its histologic features have been occasionally reported, a lack of description of the cytologic features has hampered the prompt and accurate diagnosis of this condition. Herein, we describe the cytologic findings and histology of four aspiration cytology specimens of thymic adenocarcinoma. The specimens were obtained from primary tumors, metastatic lymph nodes, and pericardial effusions. All four specimens showed three-dimensional glandular clusters with a loss of polarity and nuclear overlapping. One specimen had extensive extracellular mucinous material. Three specimens contained tumor cells with intracytoplasmic vacuoles. While the specimen with extracellular mucin showed relatively mild cytologic atypia, other specimens exhibited more atypical cytologic changes: irregular nuclear membranes, a coarse chromatin pattern, and prominent nucleoli. The cytologic features were correlated with the histologic features in each case of enteric type thymic adenocarcinoma. The differential diagnosis included other thymic carcinomas, yolk sac tumors, and metastatic adenocarcinoma from the lung or colorectum.


Subject(s)
Adenocarcinoma , Biopsy, Fine-Needle , Chromatin , Diagnosis , Diagnosis, Differential , Endodermal Sinus Tumor , Lung , Lymph Nodes , Mediastinum , Mucins , Nuclear Envelope , Pericardial Effusion , Thymoma , Thymus Gland , Vacuoles
20.
Article in Korean | WPRIM | ID: wpr-96159

ABSTRACT

Angiomyxolipoma is a recently described and rare variant of lipoma that is characterized by the proliferation of adipose tissue with a myxoid stroma and multiple thick and thin-walled blood vessels. A 51-year-old man presented with a tender subcutaneous nodule on his right arm. The tumor was completely removed by excisional biopsy, and the histopathologic features revealed a thin fibrous capsule with incomplete fibrous septa extending into the lesion, dividing it into lobules of different sizes. The fat cells were mature, with a single vacuole and an eccentric nucleus. The vascular component consisted of groups of capillaries and occasional vessels of larger caliber. Erythrocytes were present within the lumen, and myxoid stroma was found. He was diagnosed as having angiomyxolipoma. Herein, we report a rare case of subcutaneous angiomyxolipoma of the arm. We also review and discuss such cases in the literature.


Subject(s)
Humans , Middle Aged , Adipocytes , Adipose Tissue , Arm , Biopsy , Blood Vessels , Capillaries , Erythrocytes , Lipoma , Vacuoles
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