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1.
Ann Hematol ; 102(12): 3311-3323, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-37656190

RESUMEN

Hemophagocytic lymphohistiocytosis (HLH), a hyperinflammatory syndrome, is caused by the incessant activation of lymphocytes and macrophages, resulting in damage to organs, including hematopoietic organs. Recently, we demonstrated that repeated lipopolysaccharide (LPS) treatment induces HLH-like features in senescence-accelerated (SAMP1/TA-1) mice but not in senescence-resistant control (SAMR1) mice. Hematopoietic failure in LPS-treated SAMP1/TA-1 mice was attributed to hematopoietic microenvironment dysfunction, concomitant with severely imbalanced M1 and M2 macrophage polarization. Macrophages are a major component of the bone marrow (BM) hematopoietic microenvironment. Clodronate liposomes are useful tools for in vivo macrophage depletion. In this study, we depleted macrophages using clodronate liposomes to determine their role in the hematopoietic microenvironment in SAMP1/TA-1 and SAMR1 mice. Under clodronate liposome treatment, the response between SAMR1 and SAMP1/TA-1 mice differed as follows: (1) increase in the number of activated M1 and M2 macrophages derived from newly generated macrophages and M2-dominant and imbalanced M1 and M2 macrophage polarization in the BM and spleen; (2) severe anemia and thrombocytopenia; (3) high mortality rate; (4) decrease in erythroid progenitors and B cell progenitors in the BM; and (5) decrease in the mRNA expression of erythroid-positive regulators such as erythropoietin and increase in that of erythroid- and B lymphoid-negative regulators such as interferon-γ in the BM. Depletion of residual macrophages in SAMP1/TA-1 mice impaired hematopoietic homeostasis, particularly erythropoiesis and B lymphopoiesis, owing to functional impairment of the hematopoietic microenvironment accompanied by persistently imbalanced M1/M2 polarization. Thus, macrophages play a vital role in regulating the hematopoietic microenvironment to maintain homeostasis.


Asunto(s)
Linfohistiocitosis Hemofagocítica , Ratones , Animales , Linfohistiocitosis Hemofagocítica/metabolismo , Liposomas/metabolismo , Ácido Clodrónico/farmacología , Ácido Clodrónico/metabolismo , Lipopolisacáridos , Macrófagos/metabolismo
2.
Mol Cell Neurosci ; 121: 103745, 2022 07.
Artículo en Inglés | MEDLINE | ID: mdl-35660087

RESUMEN

Microgravity (MG) exposure and motor neuron diseases, such as amyotrophic lateral sclerosis (ALS), lead to motor deficits, including muscle atrophy and loss of neuronal activity. Abnormalities in motor neurons and muscles caused by MG exposure can be recovered by subsequent ground exercise. In contrast, the degeneration that occurs in ALS is irreversible. A common phenotype between MG exposure and ALS pathology is motor system abnormality, but the causes may be different. In this study, to elucidate the motor system that is affected by each condition, we investigated the effects of MG and the human SOD1 ALS mutation on gene expression in various cell types of the mouse ventral lumbar spinal cord, which is rich in motor neurons innervating the lower limb. To identify cell types affected by MG or ALS pathogenesis, we analyzed differentially expressed genes with known cell-type markers, which were determined from previous single-cell studies of the spinal cord in MG-exposed and SOD1G93A mice, an ALS mouse model. Differentially expressed genes were observed in MG mice in various spinal cord cell types, including neurons, microglia, astrocytes, oligodendrocytes, oligodendrocyte precursor cells, meningeal cells/Schwann cells, and vascular cells. We also examined neuronal populations in the spinal cord. Gene expression in putative excitatory and inhibitory neurons changed more than that in cholinergic motor neurons of the spinal cord in both MG and SOD1G93A mice. Many putative neuron types, especially visceral motor neurons, and axon initial segments (AIS) were affected in MG mice. In contrast, the effect on neurons and AIS in SOD1G93A mice was slight at P30 but progressed with aging. Interestingly, changes in dopaminergic system-related genes were specifically altered in the spinal cord of MG mice. These results indicate that MG and ALS pathology in various cell types contribute to motor neuron degeneration. Furthermore, there were more alterations in neurons in MG-exposed mice than in SOD1G93A mice. A large number of differentially expressed genes (DEGs) in MG mice represent more than SOD1G93A mice with ALS pathology. Elucidation of MG pathogenesis may provide more insight into the pathophysiology of neurodegenerative diseases.


Asunto(s)
Esclerosis Amiotrófica Lateral , Ingravidez , Esclerosis Amiotrófica Lateral/metabolismo , Animales , Modelos Animales de Enfermedad , Humanos , Ratones , Ratones Transgénicos , Neuronas Motoras/metabolismo , Médula Espinal/metabolismo , Superóxido Dismutasa/metabolismo , Superóxido Dismutasa-1/genética , Superóxido Dismutasa-1/metabolismo
3.
J Bacteriol ; 204(1): e0042021, 2022 01 18.
Artículo en Inglés | MEDLINE | ID: mdl-34694904

RESUMEN

Escherichia coli survives under acid stress conditions by the glutamic acid-dependent acid resistance (GAD) system, which enzymatically decreases intracellular protons. We found a linkage between GAD and flagellar systems in E. coli. The hdeD gene, one of the GAD cluster genes, encodes an uncharacterized membrane protein. A reporter assay showed that the hdeD promoter was induced in a GadE-dependent manner when grown in the M9 glycerol medium. Transcriptome analysis revealed that most of the transcripts were from genes involved in flagellum synthesis, and cell motility increased not only in the hdeD-deficient mutant but also in the gadE-deficient mutant. Defects in both the hdeD and gadE increased the intracellular level of FliA, an alternative sigma factor for flagellum synthesis, activated by the master regulator FlhDC. The promoter activity of the lrhA gene, which encodes repressor for the flhDC operon, was found to decrease in both the hdeD- and gadE-deficient mutants. Transmission electron microscopy showed that the number of flagellar filaments on the hdeD-, gadE-, and lrhA-deficient cells increased, and all three mutants showed higher motility than the parent strain. Thus, HdeD in the GAD system activates the lrhA promoter, resulting in a decrease in flagellar filaments in E. coli cells. We speculated that the synthesis of HdeD, stimulated in E. coli exposed to acid stress, could control the flagellum biosynthesis by sensing slight changes in pH at the cytoplasmic membrane. This could help in saving energy through termination of flagellum biosynthesis and improve bacterial survival efficiency within the animal digestive system. IMPORTANCE E. coli cells encounter various environments from the mouth down to the intestines within the host animals. The pH of gastric juice is lower than 2.0, and the bacterial must quickly respond and adapt to the following environmental changes before reaching the intestines. The quick response plays a role in cellular survival in the population, whereas adaptation may contribute to species survival. The GAD and flagellar systems are important for response to low pH in E. coli. Here, we identified the novel inner membrane regulator HdeD, encoding in the GAD cluster, to repress the synthesis of flagella. These insights provide a deeper understanding of how the bacteria enter the animal digestive system, survive, and form colonies in the intestines.


Asunto(s)
Escherichia coli K12/metabolismo , Proteínas de Escherichia coli/metabolismo , Flagelos/metabolismo , Regulación Bacteriana de la Expresión Génica/fisiología , Proteínas de la Membrana/metabolismo , Factores de Transcripción/metabolismo , Escherichia coli K12/genética , Proteínas de Escherichia coli/genética , Genoma Bacteriano , Proteínas de la Membrana/genética , Mutación , Factores de Transcripción/genética , Transcriptoma
4.
Biol Pharm Bull ; 45(11): 1602-1608, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-36328495

RESUMEN

Lipopolysaccharide (LPS) treatment induced hemophagocytic lymphohistiocytosis in senescence-accelerated mice (SAMP1/TA-1), but not in senescence-resistant control mice (SAMR1). SAMP1/TA-1 treated with LPS exhibited functional impairment of the hematopoietic microenvironment, which disrupted the dynamics of hematopoiesis. Macrophages are a major component of the bone marrow (BM) hematopoietic microenvironment, which regulates hematopoiesis. Qualitative and quantitative changes in activated macrophages in LPS-treated SAMP1/TA-1 are thought to contribute to the functional deterioration of the hematopoietic microenvironment. Thus, we examined the polarization of pro-inflammatory (M1) and anti-inflammatory (M2) macrophages, and the dynamics of macrophage production in the BM of SAMP1/TA-1 and SAMR1 after LPS treatment. After LPS treatment, the proportions of M1 and M2 macrophages and the numbers of macrophage progenitor (CFU-M) cells increased in both SAMP1/TA-1 and SAMR1. However, compared to the SAMR1, the increase in the M1 macrophage proportion was prolonged, and the increase in the M2 macrophage proportion was delayed. The increase in the number of CFU-M cells was prolonged in SAMP1/TA-1 after LPS treatment. In addition, the levels of transcripts encoding an M1 macrophage-inducing cytokine (interferon-γ) and macrophage colony-stimulating factor were markedly increased, and the increases in the levels of transcripts encoding M2 macrophage-inducing cytokines (interleukin (IL)-4, IL-10, and IL-13) were delayed in SAMP1/TA-1 when compared to SAMR1. Our results suggest that LPS treatment led to the severely imbalanced polarization of activated M1/M2 macrophages accompanied by a prolonged increase in macrophage production in the BM of SAMP1/TA-1, which led to the impairment of the hematopoietic microenvironment, and disrupted the dynamics of hematopoiesis.


Asunto(s)
Médula Ósea , Linfohistiocitosis Hemofagocítica , Ratones , Animales , Lipopolisacáridos/farmacología , Activación de Macrófagos , Macrófagos , Citocinas , Modelos Animales de Enfermedad
5.
Curr Top Microbiol Immunol ; 427: 161-172, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-31183607

RESUMEN

The flagellum is a motile organ, and the needle complex is a type III secretion apparatus for pathogenesis. There are more similarities than differences between the two structures at the molecular level. Here I focus on the hook and the needle and discuss their length control mechanism. The hook is a substructure of the flagellum and the needle is a part of the needle complex. Both structures are tubular structures that have a central channel for protein secretion. Their lengths are controlled by an intriguing mechanism involving a ruler protein and a switchable gate of the protein secretion system. A model for length control is proposed.


Asunto(s)
Sistemas de Secreción Tipo III/metabolismo , Proteínas Bacterianas , Flagelos , Transporte de Proteínas
6.
EMBO Rep ; 20(5)2019 05.
Artículo en Inglés | MEDLINE | ID: mdl-30898768

RESUMEN

Many archaea swim by means of archaella. While the archaellum is similar in function to its bacterial counterpart, its structure, composition, and evolution are fundamentally different. Archaella are related to archaeal and bacterial type IV pili. Despite recent advances, our understanding of molecular processes governing archaellum assembly and stability is still incomplete. Here, we determine the structures of Methanococcus archaella by X-ray crystallography and cryo-EM The crystal structure of Methanocaldococcus jannaschii FlaB1 is the first and only crystal structure of any archaellin to date at a resolution of 1.5 Å, which is put into biological context by a cryo-EM reconstruction from Methanococcus maripaludis archaella at 4 Å resolution created with helical single-particle analysis. Our results indicate that the archaellum is predominantly composed of FlaB1. We identify N-linked glycosylation by cryo-EM and mass spectrometry. The crystal structure reveals a highly conserved metal-binding site, which is validated by mass spectrometry and electron energy-loss spectroscopy. We show in vitro that the metal-binding site, which appears to be a widespread property of archaellin, is required for filament integrity.


Asunto(s)
Proteínas Arqueales/metabolismo , Sitios de Unión/fisiología , Metales/metabolismo , Methanococcus/metabolismo , Microscopía por Crioelectrón/métodos , Cristalografía por Rayos X , Citoesqueleto/metabolismo , Glicosilación , Espectrometría de Masas/métodos , Orgánulos/metabolismo , Dominios Proteicos/fisiología
7.
Curr Microbiol ; 78(4): 1267-1276, 2021 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-33638001

RESUMEN

The bacterium Staphylococcus aureus, which colonizes healthy human skin, may cause diseases, such as atopic dermatitis (AD). Treatment for such AD cases involves antibiotic use; however, alternate treatments are preferred owing to the development of antimicrobial resistance. This study aimed to characterize the novel bacteriophage SaGU1 as a potential agent for phage therapy to treat S. aureus infections. SaGU1 that infects S. aureus strains previously isolated from the skin of patients with AD was screened from sewage samples in Gifu, Japan. Its genome was sequenced and analyzed using bioinformatics tools, and the morphology, lytic activity, stability, and host range of the phage were determined. The SaGU1 genome was 140,909 bp with an average GC content of 30.2%. The viral chromosome contained 225 putative protein-coding genes and four tRNA genes, carrying neither toxic nor antibiotic resistance genes. Electron microscopy analysis revealed that SaGU1 belongs to the Myoviridae family. Stability tests showed that SaGU1 was heat-stable under physiological and acidic conditions. Host range testing revealed that SaGU1 can infect a broad range of S. aureus clinical isolates present on the skin of AD patients, whereas it did not kill strains of Staphylococcus epidermidis, which are symbiotic resident bacteria on human skin. Hence, our data suggest that SaGU1 is a potential candidate for developing a phage therapy to treat AD caused by pathogenic S. aureus.


Asunto(s)
Dermatitis Atópica , Staphylococcus aureus , Genoma Viral , Humanos , Japón , Fagos de Staphylococcus/genética , Staphylococcus aureus/genética
8.
Cancer Sci ; 111(6): 2132-2145, 2020 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-32304130

RESUMEN

In the cell cycle, the G1 /S transition is controlled by the cyclin-dependent kinase (CDK) 4/6-cyclin D complex. Constitutive activation of CDK4/6 dysregulates G1 /S transition, leading to oncogenic transformation. We found that 3 CDK4/6 inhibitors, abemaciclib, ribociclib, and palbociclib, exerted a cytocidal effect as well as a cytostatic effect at the G1 phase in cancer cell lines, including A549 human non-small cell lung cancer cells. Among these inhibitors, abemaciclib exhibited the most potent cytotoxic effect. The cell-death phenotype induced by abemaciclib, which entailed formation of multiple cytoplasmic vacuoles, was not consistent with apoptosis or necroptosis. Abemaciclib blocked autophagic flux, resulting in accumulation of autophagosomes, however vacuole formation and cell death induced by abemaciclib were independent of autophagy. In addition, methuosis, a cell-death phenotype characterized by vacuole formation induced by excessive macropinocytosis, was excluded because the vacuoles did not incorporate fluorescent dextran. Of note, both formation of vacuoles and induction of cell death in response to abemaciclib were inhibited by vacuolar-type ATPase (V-ATPase) inhibitors such as bafilomycin A1 and concanamycin A. Live-cell imaging revealed that the abemaciclib-induced vacuoles were derived from lysosomes that expanded following acidification. Transmission electron microscopy revealed that these vacuoles contained undigested debris and remnants of organelles. Cycloheximide chase assay revealed that lysosomal turnover was blocked by abemaciclib. Furthermore, mTORC1 inhibition along with partial lysosomal membrane permeabilization occurred after abemaciclib treatment. Together, these results indicate that, in cancer cells, abemaciclib induces a unique form of cell death accompanied by swollen and dysfunctional lysosomes.


Asunto(s)
Aminopiridinas/farmacología , Bencimidazoles/farmacología , Muerte Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Inhibidores de Proteínas Quinasas/farmacología , Antineoplásicos/farmacología , Línea Celular Tumoral , Humanos , Lisosomas/efectos de los fármacos , Vacuolas/efectos de los fármacos
9.
Ann Hematol ; 99(7): 1515-1523, 2020 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-32506245

RESUMEN

Hemophagocytic lymphohistiocytosis (HLH) is a life-threatening systemic hyperinflammatory disorder. We found recently that repeated lipopolysaccharide (LPS) treatment induces HLH-like features in senescence-accelerated mice (SAMP1/TA-1) but not in senescence-resistant control mice (SAMR1). In this study, we analyzed the dynamics of hematopoiesis in this mouse model of HLH. When treated repeatedly with LPS, the numbers of myeloid progenitor cells (CFU-GM) and B-lymphoid progenitor cells (CFU-preB) in the bone marrow (BM) rapidly decreased after each treatment in both strains. The number of CFU-GM in SAMP1/TA-1 and SAMR1, and of CFU-preB in SAMR1, returned to pretreatment levels by 7 days after each treatment. However, the recovery in the number of CFU-preB in SAMP1/TA-1 was limited. In both strains, the BM expression of genes encoding positive regulators of myelopoiesis (granulocyte colony-stimulating factor (G-CSF), granulocyte macrophage colony-stimulating factor (GM-CSF), and interleukin (IL)-6), and negative regulators of B lymphopoiesis (tumor necrosis factor (TNF)-α) was increased. The expression of genes encoding positive regulators of B lymphopoiesis (stromal-cell derived factor (SDF)-1, IL-7, and stem cell factor (SCF)) was persistently decreased in SAMP1/TA-1 but not in SAMR1. Expression of the gene encoding p16INK4a and the proportion of ß-galactosidase-positive cells were increased in cultured stromal cells obtained from LPS-treated SAMP1/TA-1 but not in those from LPS-treated SAMR1. LPS treatment induced qualitative changes in stromal cells, which comprise the microenvironment supporting appropriate hematopoiesis, in SAMP1/TA-1; these stromal cell changes are inferred to disrupt the dynamics of hematopoiesis. Thus, hematopoietic tissue is one of the organs that suffer life-threatening damage in HLH.


Asunto(s)
Médula Ósea/patología , Hematopoyesis/fisiología , Células Madre Hematopoyéticas/patología , Linfohistiocitosis Hemofagocítica/patología , Linfohistiocitosis Hemofagocítica/fisiopatología , Nicho de Células Madre/fisiología , Animales , Células de la Médula Ósea/patología , Células de la Médula Ósea/fisiología , Recuento de Células , Células Cultivadas , Microambiente Celular/fisiología , Modelos Animales de Enfermedad , Células Madre Hematopoyéticas/fisiología , Lipopolisacáridos , Linfohistiocitosis Hemofagocítica/inducido químicamente , Masculino , Ratones , Células del Estroma/patología
10.
Pain Med ; 21(5): 1018-1024, 2020 05 01.
Artículo en Inglés | MEDLINE | ID: mdl-31504919

RESUMEN

OBJECTIVE: An ultrasound-guided interfascial injection, which targets the space between the epimysia, is often performed for myofascial neck and shoulder pain. However, the relationship between the injection volume and clinical effectiveness has been controversial. We conducted an anatomical study with cadavers to measure the distribution of a small amount of pigment solution injected into the interfascial space. DESIGN: An experimental cadaveric study. SETTING: An institutional clinical anatomy laboratory. METHODS: We performed 20 ultrasound-guided injections with pigment solutions of 1.0 mL each into the space between the trapezius muscle and rhomboid muscle bilaterally on 10 cadaver specimens. Cadavers were then dissected and macroscopically evaluated for pigment distribution on the fascia of the muscular surfaces. The pigment distribution area of each injection site was visually confirmed and calculated using automatic area calculation software. RESULTS: Pigment solution exclusively within the interfascial space was visually confirmed in 95% (19/20) of injection sites. The median pigmented surface area (interquartile range) was 24.50 (16.17-30.76) cm2 on the deep side of the trapezius muscle and 18.82 (13.04-24.79) cm2 on the superficial side of rhomboid muscle; these measurements were statistically significantly different (P = 0.033). CONCLUSIONS: A pigment solution comprising as little as 1.0 mL injected under ultrasound guidance separated two adjacent muscles and spread to the wide area within the interfascial space. The difference in the pigment distribution area between the two adjacent muscles can be explained by the path made by the injection needle and several layers of fascia between the epimysium.


Asunto(s)
Músculos Superficiales de la Espalda , Cadáver , Humanos , Inyecciones , Ultrasonografía , Ultrasonografía Intervencional
11.
BMC Musculoskelet Disord ; 21(1): 322, 2020 May 22.
Artículo en Inglés | MEDLINE | ID: mdl-32443975

RESUMEN

BACKGROUND: The popliteus tendon (PT) or lateral collateral ligament (LCL) stabilizes the postero-lateral aspects of the knees. When surgeons perform total knee arthroplasty (TKA), PT and LCL iatrogenic injuries are a risk because the femoral attachments are relatively close to the femoral bone resection area. The purpose of this study was to evaluate the distance between the PT or LCL footprint and the TKA implant using a 3D template system and to evaluate any significant differences according to the implant model. METHODS: Eighteen non-paired formalin fixed cadaveric lower limbs were used (average age: 80.3). Whole length lower limbs were resected from the pelvis. All the surrounding soft tissue except the PT, knee ligaments and meniscus were removed from the limb. Careful dissection of the PT and LCL was performed, and the femoral footprints were detected. Each footprint periphery was marked with a 1.5 mm K-wire. Computed tomography (CT) scanning of the whole lower limb was then performed. The CT data was analyzed with a 3D template system. This simulation models for TKA were the Journey II BCS and the Persona PS. The area of each footprint, and the length between the most distal and posterior point of the lateral femoral condyle and the edge of each footprint were measured. Matching the implant model to the CT image of the femur, the shortest length between each footprint and the bone resection area were calculated. RESULTS: PT and LCL footprint were detected in all knees. The area of the PT and LCL footprints was 38.7 ± 17.7 mm2 and 58.0 ± 24.6 mm2, respectively. The length between the most distal and posterior point of the lateral femoral condyle and the edge of the PT footprint was 10.3 ± 2.4 mm and 14.2 ± 2.8 mm, respectively. The length between most distal and most posterior point of the lateral femoral condyle and the edge of the LCL footprint was 16.3 ± 2.3 mm and 15.5 ± 3.3 mm, respectively. Under TKA simulation, the shortest length between the PT footprint and the femoral bone resection area for the Journey II BCS and the Persona PS was 4.3 ± 2.5 mm and 3.2 ± 2.9 mm, respectively. The shortest length between the LCL footprint and the femoral bone resection area for the Journey II BCS and the Persona PS was 7.2 ± 2.3 mm and 5.6 ± 2.1 mm, respectively. The PT attachment was damaged by the bone resection of the Journey II BCS and the Persona PS TKA in 3 and 9 knees, respectively. CONCLUSION: The PT and LCL femoral attachments existed close to the femoral bone resection area of the TKA. To prevent postero-lateral instability in TKA, careful attention is needed to avoid damage to the PT and LCL during surgical procedures.


Asunto(s)
Traumatismos de la Rodilla/cirugía , Articulación de la Rodilla/cirugía , Ligamentos Laterales del Tobillo/cirugía , Traumatismos de los Tendones/cirugía , Tendones/cirugía , Anciano , Anciano de 80 o más Años , Cadáver , Femenino , Humanos , Imagenología Tridimensional , Traumatismos de la Rodilla/diagnóstico , Articulación de la Rodilla/patología , Prótesis de la Rodilla , Ligamentos Laterales del Tobillo/lesiones , Ligamentos Laterales del Tobillo/patología , Masculino , Persona de Mediana Edad , Traumatismos de los Tendones/diagnóstico , Tendones/patología
12.
Knee Surg Sports Traumatol Arthrosc ; 28(8): 2668-2674, 2020 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32157361

RESUMEN

PURPOSE: The purpose of this study was to reveal the morphological correlation between the lateral wall of femoral intercondylar notch and the Blumensaat's line. METHODS: Forty-one non-paired human cadaveric knees were included in this study (23 female, 18 male: median age 83). Knees were resected, and 3 dimensional computed tomography (3D-CT) was performed. In the axial CT image, bony protrusion (resident's ridge) and cortical thickness in the lateral wall of the femoral intercondylar notch were detected. The length between the top of the ridge, or the most anterior, middle, and most posterior border of cortical thickness and posterior femoral condylar line was measured. Following Iriuchishima's classification, the morphology of the Blumensaat's line was classified into straight and hill types (small and large hill types). In the hill types, the length between the hilltop and the posterior border of the Blumensaat's line or the posterior border of the femoral condyle was evaluated. Statistical correlation was calculated between the top of the ridge location, cortical thickness location in the notch, and hilltop location. RESULTS: There were 7 straight type knees and 34 hill type knees (9 small hill type knees and 25 large hill type knees). Only the hill types of knees were evaluated. The top of the ridge, anterior margin, middle, and posterior border of cortical thickness in the lateral wall of the femoral intercondylar notch existed at 61.8 ± 4.6%, 58.3 ± 12.3%, 42.1 ± 7.9%, and 25.5 ± 5.4% from the posterior condylar line, respectively. The hilltop existed at 24.9 ± 5.9% and 30.7 ± 5.0%, from the posterior border of the Blumensaat's line and from the posterior border of the femoral condyle, respectively. Significant correlation was observed between resident's ridge top, cortical thickness location and hilltop location. CONCLUSION: In all cadaveric knees, cortical thickness was detected in the lateral wall of the femoral intercondylar notch. The resident's ridge and cortical thickness location had significant correlation with the hill location in the Blumensaat's line, indicating a continuation of the cortical bone from the posterior cortex of the femoral shaft via the hilltop of the Blumensaat's line to the cortical thickness in the lateral wall of the femoral intercondylar notch. For clinical relevance, hilltop location in the Blumensaat's line is a new bony landmark in anterior cruciate ligament surgery.


Asunto(s)
Fémur/anatomía & histología , Articulación de la Rodilla/anatomía & histología , Anciano de 80 o más Años , Ligamento Cruzado Anterior/cirugía , Reconstrucción del Ligamento Cruzado Anterior/métodos , Cadáver , Femenino , Fémur/diagnóstico por imagen , Fémur/cirugía , Humanos , Imagenología Tridimensional , Articulación de la Rodilla/diagnóstico por imagen , Articulación de la Rodilla/cirugía , Masculino , Tomografía Computarizada por Rayos X
13.
Knee Surg Sports Traumatol Arthrosc ; 28(8): 2453-2457, 2020 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32514842

RESUMEN

PURPOSE: The purpose of this study was to evaluate the difference in the center point of the femoral ACL footprint according to the morphological variations of the Blumensaat's line. METHODS: Fifty-nine non-paired human cadaver knees were used. The ACL was cut in the middle, and the femoral bone was cut at the most proximal point of the femoral notch. Digital images were evaluated using the Image J software. The periphery of the femoral ACL footprint was outlined and the center point was measured automatically. Following Iriuchishima's classification, the morphology of the Blumensaat's line was classified into straight and hill types (small and large hill types). The center of the femoral ACL footprint and hilltop placement were evaluated using the quadrant method. A quadrant grid was placed uniformly, irregardless of hill existence, and not including the articular cartilage. A correlation analysis was performed between the center point of the femoral ACL footprint and hilltop placement. RESULTS: The straight type consisted of 19 knees, and the hill type 40 knees (small hill type 13 knees and large hill type 27 knees). The center of the femoral ACL footprint (shallow-deep/high-low) in the straight and hill type knees was 33.7/47.6%, and 37.2/50.3%, respectively. In the hill type, the ACL footprint center was significantly more shallow when compared to the straight type. Significant correlation was observed between the center point of the femoral ACL footprint and hilltop placement of the Blumensaat's line. CONCLUSION: The center point of the femoral ACL footprint was significantly more shallow in the hill type knees when compared to the straight type. For clinical relevance, considering that the location of the femoral ACL footprint center is different depending on the Blumensaat's line morphology, to perform accurate ACL reconstruction, femoral ACL tunnel placement should be made based on Blumensaat's line morphological variations.


Asunto(s)
Ligamento Cruzado Anterior/anatomía & histología , Anciano , Anciano de 80 o más Años , Ligamento Cruzado Anterior/cirugía , Reconstrucción del Ligamento Cruzado Anterior , Cadáver , Femenino , Fémur/anatomía & histología , Fémur/cirugía , Humanos , Articulación de la Rodilla/anatomía & histología , Masculino
14.
Eur J Orthop Surg Traumatol ; 30(2): 291-296, 2020 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-31552484

RESUMEN

INTRODUCTION: One of the final goals of anatomical anterior cruciate ligament (ACL) reconstruction is the restoration of native anatomy. It is essential to obtain more accurate predictors of mid-substance ACL size before surgery. However, to the best of our knowledge, no study has reported correlation between the mid-substance cross-sectional ACL size and the knee osseous morphology. The purpose of this study was to reveal correlation between the mid-substance cross-sectional ACL size and the knee osseous morphology. MATERIALS AND METHODS: We used 39 non-paired formalin fixed Japanese cadaveric knees. All surrounding muscles, ligaments and soft tissues in the knee were resected. After soft tissue resection, the knee was flexed at 90°, and a tangential plane of the femoral posterior condyles was marked and cut the ACL. Femoral ACL footprint size, Blumensaat's line length, lateral wall of the femoral intercondylar notch size, lateral wall of the femoral intercondylar notch height, tibial ACL footprint size, tibia plateau size, the whole anterior-posterior (AP) length, the medial and the lateral AP length of the tibia plateau, and the medial-lateral (ML) length of the tibia plateau were measured. The Pearson's product movement correlation was calculated to reveal correlation between the mid-substance cross-sectional ACL size and the measured parameters of the knee osseous morphology. RESULTS: The measured mid-substance cross-sectional ACL size was 49.9 ± 16.3 mm2. The tibial ACL footprint size, the tibia plateau size, the whole AP length of the tibia plateau, the lateral AP length of the tibia plateau and the ML length of the tibia plateau were significantly correlated with the mid-substance cross-sectional ACL size. CONCLUSIONS: For clinical relevance, some tibial sides of the knee osseous morphology were significantly correlated with the mid-substance cross-sectional ACL size. It might be possible to predict the mid-substance ACL size measuring these parameters.


Asunto(s)
Ligamento Cruzado Anterior/anatomía & histología , Fémur/anatomía & histología , Articulación de la Rodilla/anatomía & histología , Tibia/anatomía & histología , Anciano , Anciano de 80 o más Años , Cadáver , Epífisis/anatomía & histología , Femenino , Humanos , Masculino , Tamaño de los Órganos
15.
J Bacteriol ; 201(14)2019 07 15.
Artículo en Inglés | MEDLINE | ID: mdl-31036727

RESUMEN

The rare actinomycete Actinoplanes missouriensis produces terminal sporangia containing a few hundred flagellated spores. After release from the sporangia, the spores swim rapidly in aquatic environments as zoospores. The zoospores stop swimming and begin to germinate in niches for vegetative growth. Here, we report the characterization and functional analysis of zoospore type IV pili in A. missouriensis The pilus gene (pil) cluster, consisting of three apparently σFliA-dependent transcriptional units, is activated during sporangium formation similarly to the flagellar gene cluster, indicating that the zoospore has not only flagella but also pili. With a new method in which zoospores were fixed with glutaraldehyde to prevent pilus retraction, zoospore pili were observed relatively easily using transmission electron microscopy, showing 6 ± 3 pili per zoospore (n = 37 piliated zoospores) and a length of 0.62 ± 0.35 µm (n = 206), via observation of fliC-deleted, nonflagellated zoospores. No pili were observed in the zoospores of a prepilin-encoding pilA deletion (ΔpilA) mutant. In addition, the deletion of pilT, which encodes an ATPase predicted to be involved in pilus retraction, substantially reduced the frequency of pilus retraction. Several adhesion experiments using wild-type and ΔpilA zoospores indicated that the zoospore pili are required for the sufficient adhesion of zoospores to hydrophobic solid surfaces. Many zoospore-forming rare actinomycetes conserve the pil cluster, which indicates that the zoospore pili yield an evolutionary benefit in the adhesion of zoospores to hydrophobic materials as footholds for germination in their mycelial growth.IMPORTANCE Bacterial zoospores are interesting cells in that their physiological state changes dynamically: they are dormant in sporangia, show temporary mobility after awakening, and finally stop swimming to germinate in niches for vegetative growth. However, the cellular biology of a zoospore remains largely unknown. This study describes unprecedented zoospore type IV pili in the rare actinomycete Actinoplanes missouriensis Similar to the case for the usual bacterial type IV pili, zoospore pili appeared to be retractable. Our findings that the zoospore pili have a functional role in the adhesion of zoospores to hydrophobic solid surfaces and that the zoospores use both pili and flagella properly according to their different purposes provide an important insight into the cellular biology of the zoospore.


Asunto(s)
Actinoplanes/genética , Proteínas Fimbrias/metabolismo , Fimbrias Bacterianas/metabolismo , Esporas Bacterianas/fisiología , Actinoplanes/fisiología , Proteínas Fimbrias/genética , Fimbrias Bacterianas/genética , Regulación Bacteriana de la Expresión Génica , Esporas Bacterianas/genética
16.
Haematologica ; 104(10): 1995-2005, 2019 10.
Artículo en Inglés | MEDLINE | ID: mdl-30819910

RESUMEN

Hemophagocytic lymphohistiocytosis is a life-threatening systemic hyperinflammatory disorder with primary and secondary forms. Primary hemophagocytic lymphohistiocytosis is associated with inherited defects in various genes that affect the immunological cytolytic pathway. Secondary hemophagocytic lymphohistiocytosis is not inherited, but complicates various medical conditions including infections, autoinflammatory/autoimmune diseases, and malignancies. When senescence-accelerated mice (SAMP1/TA-1) with latent deterioration of immunological function and senescence-resistant control mice (SAMR1) were treated repeatedly with lipopolysaccharide, SAMP1/TA-1 mice displayed the clinicopathological features of hemophagocytic lymphohistiocytosis such as hepatosplenomegaly, pancytopenia, hypofibrinogenemia, hyperferritinemia, and hemophagocytosis. SAMR1 mice showed no features of hemophagocytic lymphohistiocytosis. Lipopolysaccharide induced upregulation of proinflammatory cytokines such as interleukin-1ß, interleukin-6, tumor necrosis factor-α, and interferon-γ, and interferon-γ-inducible chemokines such as c-x-c motif chemokine ligands 9 and 10 in the liver and spleen in both SAMP1/TA-1 and SAMR1 mice. However, upregulation of proinflammatory cytokines and interferon-γ-inducible chemokines in the liver persisted for longer in SAMP1/TA-1 mice than in SAMR1 mice. In addition, the magnitude of upregulation of interferon-γ in the liver and spleen after lipopolysaccharide treatment was greater in SAMP1/TA-1 mice than in SAMR1 mice. Furthermore, lipopolysaccharide treatment led to a prolonged increase in the proportion of peritoneal M1 macrophages and simultaneously to a decrease in the proportion of M2 macrophages in SAMP1/TA-1 mice compared with SAMR1 mice. Lipopolysaccharide appeared to induce a hyperinflammatory reaction and prolonged inflammation in SAMP1/TA-1 mice, resulting in features of secondary hemophagocytic lymphohistiocytosis. Thus, SAMP1/TA-1 mice represent a useful mouse model to investigate the pathogenesis of bacterial infection-associated secondary hemophagocytic lymphohistiocytosis.


Asunto(s)
Envejecimiento/inmunología , Lipopolisacáridos/toxicidad , Linfohistiocitosis Hemofagocítica , Animales , Citocinas/inmunología , Inflamación/inducido químicamente , Inflamación/inmunología , Inflamación/patología , Hígado/inmunología , Hígado/patología , Linfohistiocitosis Hemofagocítica/inducido químicamente , Linfohistiocitosis Hemofagocítica/inmunología , Linfohistiocitosis Hemofagocítica/patología , Macrófagos Peritoneales/inmunología , Macrófagos Peritoneales/patología , Ratones , Bazo/inmunología , Bazo/patología , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/inmunología
17.
Eur J Orthop Surg Traumatol ; 29(4): 849-854, 2019 May.
Artículo en Inglés | MEDLINE | ID: mdl-30701307

RESUMEN

BACKGROUNDS: "Anatomical" anterior cruciate ligament (ACL) reconstruction is defined as the functional restoration of the ACL to its native dimensions. It is essential to obtain more accurate predictors of ACL size before surgery. The purpose of this study was to investigate the correlation between the native femoral ACL footprint size and the morphology of the distal femur using three-dimensional CT (3D-CT). METHODS: Thirty non-paired Japanese human cadaver knees were used. All soft tissues around the knee were resected except the ACL. For the evaluation of femoral condyle morphology, trans-epicondylar length (TEL), notch outlet length, axial notch area, and notch width index were measured using 3D-CT. The ACL was cut in the middle, and the femoral bone was cut at the most proximal point of the femoral notch. The ACL was carefully dissected, and the boundaries of the ACL insertion site were outlined on the femoral side. An accurate lateral view of the femoral condyle was photographed with a digital camera. The size of the femoral ACL footprint, length of Blumensaat's line, and the height and area of the lateral wall of the femoral intercondylar notch were measured with ImageJ software. RESULTS: Notch height, lateral notch area, and TEL were significantly correlated with the femoral ACL footprint area. Both axial notch area and notch outlet length were significantly correlated with the femoral mid-substance insertion area. CONCLUSION: Morphological evaluation using 3D-CT preoperatively may be useful in predicting the femoral ACL footprint size.


Asunto(s)
Ligamento Cruzado Anterior/diagnóstico por imagen , Fémur/diagnóstico por imagen , Anciano , Anciano de 80 o más Años , Ligamento Cruzado Anterior/anatomía & histología , Pueblo Asiatico , Cadáver , Femenino , Fémur/anatomía & histología , Humanos , Imagenología Tridimensional , Masculino , Persona de Mediana Edad , Tomografía Computarizada por Rayos X
18.
Mol Microbiol ; 105(4): 572-588, 2017 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-28557186

RESUMEN

The bacterial flagellar export switching machinery consists of a ruler protein, FliK, and an export switch protein, FlhB and switches substrate specificity of the flagellar type III export apparatus upon completion of hook assembly. An interaction between the C-terminal domain of FliK (FliKC ) and the C-terminal cytoplasmic domain of FlhB (FlhBC ) is postulated to be responsible for this switch. FliKC has a compactly folded domain termed FliKT3S4 (residues 268-352) and an intrinsically disordered region composed of the last 53 residues, FliKCT (residues 353-405). Residues 301-350 of FliKT3S4 and the last five residues of FliKCT are critical for the switching function of FliK. FliKCT is postulated to regulate the interaction of FliKT3S4 with FlhBC , but it remains unknown how. Here we report the role of FliKCT in the export switching mechanism. Systematic deletion analyses of FliKCT revealed that residues of 351-370 are responsible for efficient switching of substrate specificity of the export apparatus. Suppressor mutant analyses showed that FliKCT coordinates FliKT3S4 action with the switching. Site-directed photo-cross-linking experiments showed that Val-302 and Ile-304 in the hydrophobic core of FliKT3S4 bind to FlhBC . We propose that FliKCT may induce conformational rearrangements of FliKT3S4 to bind to FlhBC .


Asunto(s)
Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Especificidad por Sustrato/genética , Secuencia de Aminoácidos , Transporte Biológico , Escherichia coli/genética , Escherichia coli/metabolismo , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Flagelos/metabolismo , Flagelos/microbiología , Proteínas de la Membrana/metabolismo , Dominios Proteicos , Estructura Terciaria de Proteína , Especificidad por Sustrato/fisiología
19.
Microbiology (Reading) ; 164(5): 740-750, 2018 05.
Artículo en Inglés | MEDLINE | ID: mdl-29595414

RESUMEN

Campylobacter jejuni cells have bipolar flagella. Both flagella have similar lengths of about one helical turn, or 3.53±0.52 µm. The flagellar filament is composed of two homologous flagellins: FlaA and FlaB. Mutant strains that express either FlaA or FlaB alone produce filaments that are shorter than those of the wild-type. It is reported that the flaG gene could affect filament length in some species of bacteria, but its function remains unknown. We introduced a flaG-deletion mutation into the C. jejuni wild-type strain and flaA- or flaB-deletion mutant strains, and observed their flagella by microscopy. The ΔflaG mutant cells produced long filaments of two helical turns in the wild-type background. The ΔflaAG double mutant cells produced very short FlaB filaments. On the other hand, ΔflaBG double mutant cells produced long FlaA filaments and their morphology was not helical but straight. Furthermore, FlaG was secreted, and a pulldown assay showed that sigma factor 28 was co-precipitated with purified polyhistidine-tagged FlaG. We conclude that FlaG controls flagella length by negatively regulating FlaA filament assembly and discuss the role of FlaA and FlaB flagellins in C. jejuni flagella formation.


Asunto(s)
Proteínas Bacterianas/metabolismo , Campylobacter jejuni/fisiología , Flagelos/genética , Flagelos/metabolismo , Proteínas Bacterianas/genética , Campylobacter jejuni/citología , Campylobacter jejuni/genética , Campylobacter jejuni/metabolismo , Flagelos/ultraestructura , Flagelina/genética , Flagelina/metabolismo , Eliminación de Gen , Regulación Bacteriana de la Expresión Génica , Locomoción , Microscopía Electrónica de Transmisión , Unión Proteica , Factor sigma/metabolismo
20.
Knee Surg Sports Traumatol Arthrosc ; 26(4): 1110-1116, 2018 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-28289820

RESUMEN

PURPOSE: The purpose of this study was to reveal the correlation between femoral tunnel length and the morphology of the femoral intercondylar notch in anatomical single bundle anterior cruciate ligament (ACL) reconstruction using three-dimensional computed tomography (3D-CT). METHODS: Thirty subjects undergoing anatomical single bundle ACL reconstruction were included in this study (23 female, 7 male: average age 45.5 ± 16.7). In the anatomical single bundle ACL reconstruction, the femoral and tibial tunnels were created close to the antero-medial bundle insertion site with trans-portal technique. Using post-operative three-dimensional computed tomography (3D-CT), accurate axial and lateral views of the femoral condyle were evaluated. The correlation of femoral tunnel length, which was measured intra-operatively, with the transepicondylar length (TEL), notch width index, notch outlet length, the notch area (axial), length of Blumensaat's line, and the height and area of the lateral wall of the femoral intercondylar notch was statistically analyzed. Tunnel placement was also evaluated using a Quadrant method. RESULTS: The average femoral tunnel length was 35.4 ± 4.4 mm. The average TEL, NWI, notch outlet length, and the axial notch area, were 76.9 ± 5.1 mm, 29.1 ± 3.8%, 19.5 ± 3.9 mm, and 257.4 ± 77.4 mm2, respectively. The length of Blumensaat's line and the height and area of the lateral wall of the femoral intercondylar notch were 33.8 ± 3.2 mm, 22.8 ± 2.3 mm, and 738.7 ± 129 mm2, respectively. The length of Blumensaat's line, the height, and the area of the lateral wall of the femoral intercondylar notch were significantly correlated with femoral tunnel length. Femoral tunnel placement was 23.4 ± 4.5% in a shallow-deep direction and 35.4 ± 8.8% in a high-low direction. CONCLUSION: The length of Blumensaat's line, height, and area of the lateral wall of the femoral intercondylar notch are correlated with femoral tunnel length in anatomical single bundle ACL reconstruction. For clinical relevance, these parameters are useful in predicting the length of the femoral tunnel in anatomical single bundle ACL reconstruction for the prevention of extremely short femoral tunnel creation. LEVEL OF EVIDENCE: Case controlled study, Level III.


Asunto(s)
Lesiones del Ligamento Cruzado Anterior/cirugía , Reconstrucción del Ligamento Cruzado Anterior/métodos , Fémur/anatomía & histología , Articulación de la Rodilla/anatomía & histología , Tomografía Computarizada por Rayos X , Adulto , Femenino , Fémur/diagnóstico por imagen , Fémur/cirugía , Humanos , Imagenología Tridimensional , Articulación de la Rodilla/diagnóstico por imagen , Articulación de la Rodilla/cirugía , Masculino , Persona de Mediana Edad
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