RESUMEN
Vibrio alginolyticus, a gram-negative marine bacterium, poses significant health risks through various infections transmitted via contaminated seawater or seafood consumption. This case report details a 42-year-old male presenting with chronic seropurulent discharge from his ear, ultimately diagnosed with otitis externa caused by V. alginolyticus. Examination findings and antibiotic sensitivity testing informed the treatment strategy, leading to a successful resolution. The increasing incidence of V. alginolyticus infections, particularly in warm coastal water, necessitated heightened clinical awareness and appropriate management. As global temperatures rise, proactive measures including patient education and accurate diagnosis become crucial in preventing disease progression and complications associated with V. alginolyticus infections.
RESUMEN
INTRODUCTION: Lack of physical activity (PA) is associated with obesity, diabetes, hypertension, cardiovascular diseases, and cancer. Parenting practices influence PA in young children. However, there is little evidence available for adolescents. We examined whether parenting practices were associated with out-of-school PA (OSPA) in US adolescents. METHODS: This cross-sectional 2019 study analyzed data from the 2014 FLASHE study, a web-based, quota-sampled survey of parent-adolescent dyads. Inclusion required survey completion and parents to live with their teen (ages 12-17 y old). Physically limited adolescents were excluded. Dyads were stratified by teen age. Exposures included parental modeling, monitoring, facilitation, restriction, guided choice, and pressure. The outcomes of interest were OSPA Youth Activity Profile scores. Odds ratios (ORs) with 95% confidence intervals (CI) were calculated using adjusted logistic regressions. RESULTS: A total of 1109 dyads were included. Guided choice increased odds of OSPA for 15- to 17-year-olds (OR = 2.12; 95% CI, 1.17-3.84). Facilitation increased odds of OSPA for 12- to 14-year-olds (OR = 2.21; 95% CI, 1.13-4.33). Monitoring decreased odds of OSPA for 15- to 17-year-olds (OR = 0.34; 95% CI, 0.20-0.57) and 12- to 14-year-olds (OR = 0.45; 95% CI, 0.27-0.74). Friend support increased odds of OSPA in 15- to 17-year-olds (OR = 4.03; 95% CI, 2.29-7.08) and 12- to 14-year-olds (OR = 3.05; 95% CI 1.69-5.51). CONCLUSION: Future interventions should prioritize (1) shared decision making for older teens, (2) access to PA opportunities for younger adolescents, and (3) promoting peer PA and friend support for everyone.
Asunto(s)
Ejercicio Físico , Responsabilidad Parental , Humanos , Adolescente , Masculino , Femenino , Estudios Transversales , Responsabilidad Parental/psicología , Niño , Estados Unidos , Encuestas y Cuestionarios , Relaciones Padres-HijoRESUMEN
AIMS AND OBJECTIVES: Since the advent of direct acting antiviral agents, there is a revolutionary change in the management of HCV infection. Newer drugs with different mechanism of action are being introduced and are expected to be available in coming few months in Pakistan as well. The main purpose of the guideline is to review and induct the latest research in field of HCV infection in Pakistani perspective so that our healthcare professionals can apply the new recommendations in timely and judicial manner. Target groups of guidelines are general physicians treating hepatitis C, hepatologists and gastroenterologists. Other beneficiaries of these guidelines are public health institutions of Pakistan, which provide free treatment to deserving patients under National Hepatitis Prevention and Control Program and Pakistan Bait-ul- Mal Program. METHODOLOGY: These guidelines are based on the review of National consensus practice guidelines: Diagnosis, Management and Prevention of Hepatitis C Pakistan 2009. Published data in National and International Journals searched with the help of Google search and pub med, and 2015-16 guidelines of HCV by AASLD, EASL, APASL and WHO. Local studies are preferably added with references to enhance the Pakistani perspective. Evidence was also taken from published studies. Recommendations have been based upon evidence from national publications on the subject and scientific presentations at national liver meeting as well from experts' personal experience and opinion.
Asunto(s)
Hepatitis C/diagnóstico , Hepatitis C/terapia , Antivirales/uso terapéutico , Control de Enfermedades Transmisibles , Genotipo , Hepacivirus/genética , Hepatitis C/epidemiología , Humanos , Tamizaje Masivo , Exposición Profesional/prevención & control , Pakistán/epidemiología , PrevalenciaRESUMEN
Our previous studies indicate that Secreted Protein Acidic and Rich in Cysteine (SPARC) expression suppressed medulloblastoma tumor growth in vitro and in vivo. Here we sought to determine the effect of SPARC expression in medulloblastoma cells to chemotherapeutic agents. In this study, we show that SPARC expression induces cisplatin resistance in medulloblastoma cells. We also demonstrate that the autophagy was involved in SPARC expression mediated resistance to cisplatin. Suppression of autophagy by either autophagy inhibitor, 3-methyladenosine (3MA) or Atg5 siRNA enhanced cisplatin sensitivity in SPARC expressed cells. Further, SPARC expression suppressed miR-let-7f-1 expression which resulted in disrupted repression of High Mobility Group Box 1 (HMGB1), a critical regulator of autophagy. We also show that HMGB1 is a direct target of miR-let-7f-1 and forced expression of HMGB1 cDNA enhanced cisplatin sensitivity in SPARC expressed cells. In summary, our results suggest that SPARC modulates cisplatin resistance by modulating the Let-7f-1 miRNA/HMGB1 axis in medulloblastoma cells.
Asunto(s)
MicroARNs/metabolismo , Osteonectina/metabolismo , Adenosina/análogos & derivados , Adenosina/farmacología , Antineoplásicos/toxicidad , Autofagia/efectos de los fármacos , Proteína 5 Relacionada con la Autofagia , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Cisplatino/toxicidad , Resistencia a Antineoplásicos , Proteína HMGB1/genética , Proteína HMGB1/metabolismo , Humanos , Meduloblastoma/metabolismo , Meduloblastoma/patología , MicroARNs/genética , Proteínas Asociadas a Microtúbulos/antagonistas & inhibidores , Proteínas Asociadas a Microtúbulos/genética , Proteínas Asociadas a Microtúbulos/metabolismo , Osteonectina/genética , Interferencia de ARN , ARN Interferente Pequeño/metabolismoRESUMEN
Heme is critical for a variety of cellular processes, but excess intracellular heme may result in oxidative stress and membrane injury. Feline leukemia virus subgroup C receptor (FLVCR1), a member of the SLC49 family of four paralogous genes, is a cell surface heme exporter, essential for erythropoiesis and systemic iron homeostasis. Disruption of FLVCR1 function blocks development of erythroid progenitors, likely due to heme toxicity. Mutations of SLC49A1 encoding FLVCR1 are noted in patients with a rare neurodegenerative disorder: posterior column ataxia with retinitis pigmentosa. FLVCR2 is highly homologous to FLVCR1 and may function as a cellular heme importer. Mutations of SLC49A2 encoding FLVCR2 are observed in Fowler syndrome, a rare proliferative vascular disorder of the brain. The functions of the remaining members of the SLC49 family, MFSD7 and DIRC2 (encoded by the SLC49A3 and SLC49A4 genes), are unknown, although the latter is implicated in hereditary renal carcinomas. SLC48A1 (heme responsive gene-1, HRG-1), the sole member of the SLC48 family, is associated with the endosome and appears to transport heme from the endosome into the cytosol.
Asunto(s)
Hemo/metabolismo , Hemoproteínas/metabolismo , Proteínas de Transporte de Membrana/genética , Proteínas de Transporte de Membrana/fisiología , Modelos Moleculares , Familia de Multigenes , Receptores Virales/genética , Receptores Virales/fisiología , Trastornos Cerebrovasculares/genética , Endosomas/metabolismo , Células Precursoras Eritroides/fisiología , Regulación de la Expresión Génica/fisiología , Homeostasis/fisiología , Humanos , Hierro/metabolismo , Proteínas de Transporte de Membrana/metabolismo , Mutación/genética , Receptores Virales/metabolismoRESUMEN
The pattern of disassembly of the cytoskeletal network of murine erythrocytes with deficiency of either dematin-headpiece or ß-adducin or both proteins were investigated using atomic force microscopy. A heterogeneous complex structure with fine filament features and coarse features was observed in the cytoskeleton of wild type mouse erythrocytes, whereas a significant amount of rearrangement and aggregation occurred in the mutants, particularly in the cells carrying double gene mutations. These results are consistent with the cellular and biochemical phenotype of the mutant cell membranes as being more fragile due to weakened vertical connections with the plasma membrane.
Asunto(s)
Proteínas de Unión a Calmodulina/deficiencia , Proteínas del Citoesqueleto/deficiencia , Citoesqueleto/metabolismo , Citoesqueleto/ultraestructura , Eritrocitos/metabolismo , Eritrocitos/ultraestructura , Microscopía de Fuerza Atómica/métodos , Animales , RatonesRESUMEN
Heme serves as a co-factor in proteins involved in fundamental biological processes including oxidative metabolism, oxygen storage and transport, signal transduction and drug metabolism. In addition, heme is important for systemic iron homeostasis in mammals. Heme has important regulatory roles in cell biology, yet excessive levels of intracellular heme are toxic; thus, mechanisms have evolved to control the acquisition, synthesis, catabolism and expulsion of cellular heme. Recently, a number of transporters of heme and heme synthesis intermediates have been described. Here we review aspects of heme metabolism and discuss our current understanding of heme transporters, with emphasis on the function of the cell-surface heme exporter, FLVCR. Knockdown of Flvcr in mice leads to both defective erythropoiesis and disturbed systemic iron homeostasis, underscoring the critical role of heme transporters in mammalian physiology. This article is part of a Special Issue entitled: 11th European Symposium on Calcium.
Asunto(s)
Hemo Oxigenasa (Desciclizante)/metabolismo , Hemo/metabolismo , Espacio Intracelular/metabolismo , Proteínas de Transporte de Membrana/metabolismo , Absorción , Animales , Transporte Biológico , Hemo/biosíntesis , HumanosAsunto(s)
Membrana Eritrocítica/metabolismo , Membrana Eritrocítica/fisiología , Eritrocitos/patología , Proteínas de Unión al GTP/fisiología , Enfermedades Hematológicas/metabolismo , Transglutaminasas/fisiología , Apoptosis/genética , Apoptosis/fisiología , Supervivencia Celular/genética , Deformación Eritrocítica/genética , Deformación Eritrocítica/fisiología , Membrana Eritrocítica/ultraestructura , Proteínas de Unión al GTP/genética , Proteínas de Unión al GTP/metabolismo , Enfermedades Hematológicas/genética , Enfermedades Hematológicas/patología , Humanos , Fluidez de la Membrana/genética , Fluidez de la Membrana/fisiología , Modelos Biológicos , Proteína Glutamina Gamma Glutamiltransferasa 2 , Transglutaminasas/genética , Transglutaminasas/metabolismoRESUMEN
As mediators of innate immunity, neutrophils respond to chemoattractants by adopting a highly polarized morphology. Efficient chemotaxis requires the formation of one prominent pseudopod at the cell front characterized by actin polymerization, while local inhibition suppresses the formation of rear and lateral protrusions. This asymmetric control of signaling pathways is required for directional migration along a chemotactic gradient. Here, we identify the MAGUK protein p55/MPP1 as a mediator of the frontness signal required for neutrophil polarization. We developed a p55 knockout (p55(-/-)) mouse model, and demonstrate that p55(-/-) neutrophils form multiple transient pseudopods upon chemotactic stimulation, and do not migrate efficiently in vitro. Upon agonist stimulation, p55 is rapidly recruited to the leading edge of neutrophils in mice and humans. Total F-actin polymerization, along with Rac1 and RhoA activation, appear to be normal in p55(-/-) neutrophils. Importantly, phosphorylation of Akt is significantly decreased in p55(-/-) neutrophils upon chemotactic stimulation. The activity of immunoprecipitated phosphatidylinositol 3-kinase gamma (PI3Kgamma), responsible for chemoattractant-induced synthesis of PIP(3) and Akt phosphorylation, is unperturbed in p55(-/-) neutrophils. Although the total amount of PIP(3) is normal in p55(-/-) neutrophils, PIP(3) is diffusely localized and forms punctate aggregates in activated p55(-/-) neutrophils, as compared to its accumulation at the leading edge membrane in the wild type neutrophils. Together, these results show that p55 is required for neutrophil polarization by regulating Akt phosphorylation through a mechanism that is independent of PI3Kgamma activity.
Asunto(s)
Polaridad Celular , Guanilato-Quinasas/metabolismo , Neutrófilos , Actinas/metabolismo , Animales , Quimiotaxis de Leucocito , Fosfatidilinositol 3-Quinasa Clase Ib , Células Madre Embrionarias/citología , Células Madre Embrionarias/fisiología , Activación Enzimática , Femenino , GTP Fosfohidrolasas/metabolismo , Guanilato-Quinasas/genética , Humanos , Inositol 1,4,5-Trifosfato/metabolismo , Isoenzimas/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Neuropéptidos/metabolismo , Neutrófilos/citología , Neutrófilos/metabolismo , Fenotipo , Fosfatidilinositol 3-Quinasas/metabolismo , Embarazo , Proteínas Proto-Oncogénicas c-akt/metabolismo , Trasplante de Células Madre , Proteínas de Unión al GTP rac/metabolismo , Proteína de Unión al GTP rac1 , Proteína de Unión al GTP rhoA/metabolismoRESUMEN
Dematin is an actin-binding protein originally identified in the junctional complex of the erythrocyte plasma membrane, and is present in many nonerythroid cells. Dematin headpiece knockout mice display a spherical red cell phenotype and develop a compensated anemia. Dematin has two domains: a 315-residue, proline-rich "core" domain and a 68-residue carboxyl-terminal villin-type "headpiece" domain. Expression of full-length dematin in E. coli as a GST recombinant protein results in truncation within a proline, glutamic acid, serine, threonine rich region (PEST). Therefore, we designed a mutant construct that replaces the PEST sequence. The modified dematin has high actin binding activity as determined by actin sedimentation assays. Negative stain electron microscopy demonstrates that the modified dematin also exhibits actin bundling activity like that of native dematin. Circular dichroism (CD) and NMR spectral analysis, however, show little secondary structure in the modified dematin. The lack of secondary structure is also observed in native dematin purified from human red blood cells. (15)N-HSQC NMR spectra of modified dematin indicate that the headpiece domain is fully folded whereas the core region is primarily unfolded. Our finding suggests that the core is natively unfolded and may serve as a scaffold to organize the components of the junctional complex.
Asunto(s)
Proteínas Sanguíneas/química , Fosfoproteínas/química , Pliegue de Proteína , Citoesqueleto de Actina/metabolismo , Actinas/metabolismo , Secuencia de Aminoácidos , Proteínas Sanguíneas/genética , Proteínas Sanguíneas/metabolismo , Escherichia coli/genética , Humanos , Proteínas de Microfilamentos , Datos de Secuencia Molecular , Resonancia Magnética Nuclear Biomolecular , Fosfoproteínas/genética , Fosfoproteínas/metabolismo , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Resonancia por Plasmón de SuperficieRESUMEN
Neurofibromatosis type 2 is an inherited disorder characterized by the development of benign and malignant tumors on the auditory nerves and central nervous system with symptoms including hearing loss, poor balance, skin lesions, and cataracts. Here, we report a novel protein-protein interaction between NF2 protein (merlin or schwannomin) and erythrocyte p55, also designated as MPP1. The p55 is a conserved scaffolding protein with postulated functions in cell shape, hair cell development, and neural patterning of the retina. The FERM domain of NF2 protein binds directly to p55, and surface plasmon resonance analysis indicates a specific interaction with a kD value of 3.7 nM. We developed a specific monoclonal antibody against human erythrocyte p55, and found that both p55 and NF2 proteins are colocalized in the non-myelin-forming Schwann cells. This finding suggests that the p55-NF2 protein interaction may play a functional role in the regulation of apico-basal polarity and tumor suppression pathways in non-erythroid cells.
Asunto(s)
Proteínas Sanguíneas/metabolismo , Proteínas de la Membrana/metabolismo , Neurofibromina 2/metabolismo , Animales , Anticuerpos Monoclonales/biosíntesis , Proteínas Sanguíneas/química , Humanos , Inmunohistoquímica , Proteínas de la Membrana/química , Ratones , Vaina de Mielina/metabolismo , Neuronas/metabolismo , Neuronas/patología , Unión Proteica , Estructura Terciaria de Proteína , Transporte de Proteínas , Ratas , Células de Schwann/metabolismo , Resonancia por Plasmón de SuperficieRESUMEN
Direct physical linkage of MAGUKs to the actin cytoskeleton was first established by the interaction of erythrocyte p55 with the FERM domain of protein 4.1R. Subsequently, it was reported that p55 binds to a 51-amino acid peptide, encoded by exon 10, located within the FERM domain of protein 4.1R. In this study, we investigated the nature of the p55-FERM domain binding interface and show that p55 binds to a second 35-amino acid peptide, encoded by an alternatively spliced exon 5, located within the FERM domain of protein 4.1R. Competition and Surface Plasmon Resonance-binding measurements suggest that the peptides encoded by exons 5 and 10 bind to independent sites within the D5 domain of p55. Interestingly, the full length 135 kDa isoform of protein 4.1R containing both exons 5 and 10 was targeted exclusively to the plasma membrane of epithelial cells whereas the same isoform without exon 5 completely lost its membrane localization capacity. Together, these results indicate that p55 binds to two distinct sites within the FERM domain, and the alternatively spliced exon 5 is necessary for the membrane targeting of protein 4.1R in epithelial cells. Since sequences similar to the exon 5-peptide of protein 4.1R and D5 domain of p55 are conserved in many proteins, our findings suggest that a similar mechanism may govern the membrane targeting of other FERM domain containing proteins.
Asunto(s)
Empalme Alternativo/genética , Proteínas Sanguíneas/metabolismo , Membrana Celular/metabolismo , Proteínas del Citoesqueleto/química , Proteínas del Citoesqueleto/genética , Células Epiteliales/metabolismo , Exones/genética , Proteínas de la Membrana/química , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Animales , Sitios de Unión , Unión Competitiva , Perros , Células Epiteliales/citología , Humanos , Modelos Biológicos , Péptidos/metabolismo , Unión Proteica , Isoformas de Proteínas/química , Isoformas de Proteínas/metabolismo , Estructura Terciaria de Proteína , Transporte de ProteínasRESUMEN
Plasmodium falciparum uses multiple host receptors to attach and invade human erythrocytes. Glycophorins have been implicated as receptors for parasite invasion in human erythrocytes. Here, we screened a phage display cDNA library of P. falciparum (FCR3, a sialic acid-dependent strain) using purified glycophorins and erythrocytes as bait. Several phage clones were identified that bound to immobilized glycophorins and contained the same 74 bp insert encoding the 7-amino acids sequence ETTLKSF. A similar screen using intact human erythrocytes in solution identified additional phage clones containing the same 7-amino acids sequence. Using ELISA and immunofluorescence, direct binding of ETTLKSF peptide to glycophorins and erythrocytes was confirmed. Pull-down and protease treatment assays suggest that ETTLKSF peptide specifically interacts with glycophorin C. The synthetic ETTLKSF peptide partially blocks merozoite invasion in human erythrocytes. Further characterization of ETTLKSF peptide could lead to the development of a novel class of inhibitors against the blood stage malaria.
Asunto(s)
Antimaláricos/farmacología , Malaria Falciparum/parasitología , Oligopéptidos/farmacología , Plasmodium falciparum/efectos de los fármacos , Proteínas Protozoarias/farmacología , Secuencia de Aminoácidos , Animales , Antimaláricos/química , Antimaláricos/aislamiento & purificación , Células Cultivadas , Eritrocitos/parasitología , Glicoforinas/química , Humanos , Malaria Falciparum/sangre , Oligopéptidos/química , Oligopéptidos/genética , Oligopéptidos/aislamiento & purificación , Biblioteca de Péptidos , Proteínas Protozoarias/química , Proteínas Protozoarias/genética , Proteínas Protozoarias/aislamiento & purificaciónRESUMEN
Dematin and adducin are actin-binding proteins located at the spectrin-actin junctions, also called the junctional complex, in the erythrocyte membrane. Here we propose a new model whereby dematin and adducin link the junctional complex to human erythrocyte plasma membrane. Using a combination of surface labeling, immunoprecipitation, and vesicle proteomics approaches, we have identified glucose transporter-1 as the receptor for dematin and adducin in the human erythrocyte membrane. This finding is the first description of a transmembrane protein that binds to dematin and adducin, thus providing a rationale for the attachment of the junctional complex to the lipid bilayer. Because homologues of dematin, adducin, and glucose transporter-1 exist in many non-erythroid cells, we propose that a conserved mechanism may exist that couples sugar and other related transporters to the actin cytoskeleton.
Asunto(s)
Proteínas Sanguíneas/metabolismo , Proteínas de Unión a Calmodulina/metabolismo , Citoesqueleto/metabolismo , Membrana Eritrocítica/metabolismo , Transportador de Glucosa de Tipo 1/metabolismo , Animales , Proteínas Sanguíneas/genética , Línea Celular , Transportador de Glucosa de Tipo 1/genética , Humanos , Ratones , Unión Proteica , ProteómicaRESUMEN
Dematin and adducin are actin-binding proteins of the erythrocyte "junctional complex." Individually, they exert modest effects on erythrocyte shape and membrane stability, and their homologues are expressed widely in non-erythroid cells. Here we report generation and characterization of double knock-out mice lacking beta-adducin and the headpiece domain of dematin. The combined mutations result in altered erythrocyte morphology, increased membrane instability, and severe hemolysis. Peripheral blood analysis shows evidence of severe hemolytic anemia with reduced number of erythrocytes/hematocrit/hemoglobin and an approximately 12-fold increase in the number of circulating reticulocytes. The presence of a variety of misshapen and fragmented erythrocytes correlates with increased osmotic fragility and reduced in vivo life span. Despite the apparently normal protein composition of the mutant erythrocyte membrane, the retention of the spectrin-actin complex in the membrane under low ionic strength conditions is significantly reduced by the double mutation. Atomic force microscopy reveals an increase in grain size and a decrease in filament number of the mutant membrane cytoskeleton, although the volume parameter is similar to wild type erythrocytes. Aggregated, disassembled, and irregular features are visualized in the mutant membrane, consistent with the presence of large protein aggregates. Importantly, purified dematin binds to the stripped inside-out vesicles in a saturable manner, and dematin-membrane binding is abolished upon pretreatment of membrane vesicles with trypsin. Together, these results reveal an essential role of dematin and adducin in the maintenance of erythrocyte shape and membrane stability, and they suggest that the dematin-membrane interaction could link the junctional complex to the plasma membrane in erythroid cells.
Asunto(s)
Actinas/metabolismo , Anemia Hemolítica/sangre , Proteínas Sanguíneas/genética , Proteínas de Unión a Calmodulina/genética , Membrana Eritrocítica/patología , Eliminación de Gen , Fragilidad Osmótica/genética , Fosfoproteínas/genética , Espectrina/metabolismo , Actinas/fisiología , Anemia Hemolítica/genética , Animales , Proteínas Sanguíneas/deficiencia , Proteínas Sanguíneas/metabolismo , Proteínas Sanguíneas/fisiología , Proteínas de Unión a Calmodulina/sangre , Proteínas de Unión a Calmodulina/deficiencia , Proteínas del Citoesqueleto , Modelos Animales de Enfermedad , Membrana Eritrocítica/metabolismo , Membrana Eritrocítica/ultraestructura , Humanos , Ratones , Ratones Noqueados , Microscopía de Fuerza Atómica , Fosfoproteínas/deficiencia , Fosfoproteínas/metabolismo , Fosfoproteínas/fisiología , Espectrina/fisiologíaRESUMEN
Plant pathogenesis-related proteins, including beta-1,3-glucanses, are believed to be involved in plant defense mechanisms. We have cloned a beta-1,3-glucanse gene from strawberry (Fragaria x ananassa Duch). This gene, designated FaBG2-1, is a Class II glucanase gene composed of two exons and one intron. The location of a 397-base intron in the gene was confirmed by sequencing a partial cDNA clone obtained by using a rapid amplification of cDNA ends procedure. Also, based on the cDNA sequence, the transcription start site of FaBG2-1 was assigned to a -54G residue. Genomic Southern blot analysis indicated that FaBG2-1 is a member of a multi-gene family. Reverse transcriptase-polymerase chain reaction analysis revealed that this beta-1,3-glucanase gene is expressed constitutively in strawberry leaves.