RESUMEN
Vibrio cholerae O1 is a natural inhabitant of aquatic environments and causes the diarrheal disease, cholera. Two of its primary virulence regulators, TcpP and ToxR, are localized in the inner membrane. TcpP is encoded on the Vibrio Pathogenicity Island (VPI), a horizontally acquired mobile genetic element, and functions primarily in virulence gene regulation. TcpP has been shown to undergo regulated intramembrane proteolysis (RIP) in response to environmental conditions that are unfavorable for virulence gene expression. ToxR is encoded in the ancestral genome and is present in non-pathogenic strains of V. cholerae, indicating it has roles outside of the human host. In this study, we show that ToxR undergoes RIP in V. cholerae in response to nutrient limitation at alkaline pH, a condition that occurs during the stationary phase of growth. This process involves the site-2 protease RseP (YaeL), and is dependent upon the RpoE-mediated periplasmic stress response, as deletion mutants for the genes encoding these two proteins cannot proteolyze ToxR under nutrient limitation at alkaline pH. We determined that the loss of ToxR, genetically or by proteolysis, is associated with entry of V. cholerae into a dormant state in which the bacterium is normally found in the aquatic environment called viable but nonculturable (VBNC). Strains that can proteolyze ToxR, or do not encode it, lose culturability, experience a change in morphology associated with cells in VBNC, yet remain viable under nutrient limitation at alkaline pH. On the other hand, mutant strains that cannot proteolyze ToxR remain culturable and maintain the morphology of cells in an active state of growth. Overall, our findings provide a link between the proteolysis of a virulence regulator and the entry of a pathogen into an environmentally persistent state.
Asunto(s)
Proteínas Bacterianas/metabolismo , Proteínas de Unión al ADN/metabolismo , Proteolisis , Factores de Transcripción/metabolismo , Vibrio cholerae/patogenicidad , División Celular , Endopeptidasas/genética , Endopeptidasas/metabolismo , Mutación , Vibrio cholerae/crecimiento & desarrollo , Vibrio cholerae/metabolismoRESUMEN
BACKGROUND: Leukocyte esterase (LE) was recently reported to be an accurate marker for diagnosing periprosthetic joint infection (PJI) as defined by the Musculoskeletal Infection Society (MSIS) criteria. However, the diagnostic value of the LE test for PJI after total knee arthroplasty (TKA), the reliability of the subjective visual interpretation of the LE test, and the correlation between the LE test results and the current MSIS criteria remain unclear. METHODS: This study prospectively enrolled 60 patients undergoing revision TKA for either PJI or aseptic failure. Serological marker, synovial fluid, and histological analyses were performed in all cases. The PJI group comprised 38 cases that met the MSIS criteria and the other 22 cases formed the aseptic group. All the LE tests were interpreted using both visual judgment and automated colorimetric reader. RESULTS: When "++" results were considered to indicate a positive PJI, the sensitivity, specificity, positive and negative predictive value, and diagnostic accuracy were 84, 100, 100, 79, and 90%, respectively. The visual interpretation agreed with the automated colorimetric reader in 90% of cases (Cronbach α = 0.894). The grade of the LE test was strongly correlated with the synovial white blood cell count (ρ = 0.695) and polymorphonuclear leukocyte percentage (ρ = 0.638) and moderately correlated with the serum C-reactive protein and erythrocyte sedimentation rate. CONCLUSION: The LE test has high diagnostic value for diagnosing PJI after TKA. Subjective visual interpretation of the LE test was reliable and valid for the current battery of PJI diagnostic tests according to the MSIS criteria.
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Artritis Infecciosa/diagnóstico , Artroplastia de Reemplazo de Rodilla/efectos adversos , Hidrolasas de Éster Carboxílico/análisis , Infecciones Relacionadas con Prótesis/diagnóstico , Líquido Sinovial/química , Anciano , Anciano de 80 o más Años , Artritis Infecciosa/etiología , Biomarcadores/análisis , Sedimentación Sanguínea , Proteína C-Reactiva/análisis , Colorimetría , Femenino , Humanos , Recuento de Leucocitos , Masculino , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Estudios Prospectivos , Infecciones Relacionadas con Prótesis/etiología , Tiras Reactivas , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
BACKGROUND: Intraoperative anaphylaxis to neuromuscular blocking agents (NMBAs) is a rare event that is unpredictable and potentially life threatening. Most of the previous reports on such intraoperative anaphylaxis used market share surveys or self-reported data to estimate the incidence. OBJECTIVE: To determine the incidences of intraoperative anaphylaxis to NMBAs using electronic medical records. DESIGN: A retrospective observational study. SETTING: Two tertiary hospitals in South Korea. PATIENTS: This study involved patients exposed to NMBAs during anaesthesia between 1 January 2005 and 31 May 2014. Nineteen episodes were deemed to be intraoperative anaphylaxis to NMBAs. MAIN OUTCOME MEASURES: We calculated the incidences of intraoperative anaphylaxis to NMBAs. Exposure to the agents was determined from intraoperative records maintained in an electronic medical recording system. An anaphylactic reaction was determined from both clinical signs and the results of skin tests. RESULTS: Over 9 years, 729â429 patients were exposed to NMBA, the most frequently used being rocuronium [425â047 (58.3%)] and vecuronium [274â801 (37.7%)]. The overall incidence of intraoperative anaphylaxis was 2.6 per 100â000 (19 cases), and was higher with rocuronium (16 cases, 3.8 per 100â000) than with vecuronium (two cases, 0.7 cases per 100â000), Pâ=â0.014. Comparing the first 3 years with the last 6 years, the incidence of intraoperative rocuronium anaphylaxis appeared to increase 1.4-fold (Pâ=â0.006). CONCLUSION: Among commonly used NMBAs, rocuronium appears to have the highest incidence of anaphylaxis. Our findings suggest that future prospective investigation for NMBA-induced anaphylaxis should use internationally agreed skin test protocols.
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Anafilaxia/inducido químicamente , Anafilaxia/epidemiología , Androstanoles/efectos adversos , Cuidados Intraoperatorios/efectos adversos , Bloqueo Neuromuscular/efectos adversos , Fármacos Neuromusculares no Despolarizantes/efectos adversos , Adolescente , Adulto , Anciano , Anafilaxia/diagnóstico , Registros Electrónicos de Salud , Femenino , Humanos , Incidencia , Masculino , Persona de Mediana Edad , República de Corea/epidemiología , Estudios Retrospectivos , Factores de Riesgo , Rocuronio , Pruebas Cutáneas , Centros de Atención Terciaria , Factores de Tiempo , Bromuro de Vecuronio/efectos adversosRESUMEN
BACKGROUND: Multiple tick saliva proteins, the majority of which are unknown, confer tick resistance in repeatedly infested animals. The objective of this study was to identify the 24-48 h fed Amblyomma americanum tick saliva immuno-proteome. The 24-48 h tick-feeding phase is critical to tick parasitism as it precedes important events in tick biology, blood meal feeding and disease agent transmission. Fed male, 24 and 96 h fed female phage display cDNA expression libraries were biopanned using rabbit antibodies to 24 and 48 h fed A. americanum female tick saliva proteins. Biopanned immuno-cDNA libraries were subjected to next generation sequencing, de novo assembly, and bioinformatic analysis. RESULTS: More than 800 transcripts that code for 24-48 h fed A. americanum immuno-proteins are described. Of the 895 immuno-proteins, 52% (464/895) were provisionally identified based on matches in GenBank. Of these, ~19% (86/464) show high level of identity to other tick hypothetical proteins, and the rest include putative proteases (serine, cysteine, leukotriene A-4 hydrolase, carboxypeptidases, and metalloproteases), protease inhibitors (serine and cysteine protease inhibitors, tick carboxypeptidase inhibitor), and transporters and/or ligand binding proteins (histamine binding/lipocalin, fatty acid binding, calreticulin, hemelipoprotein, IgG binding protein, ferritin, insulin-like growth factor binding proteins, and evasin). Others include enzymes (glutathione transferase, cytochrome oxidase, protein disulfide isomerase), ribosomal proteins, and those of miscellaneous functions (histamine release factor, selenoproteins, tetraspanin, defensin, heat shock proteins). CONCLUSIONS: Data here demonstrate that A. americanum secretes a complex cocktail of immunogenic tick saliva proteins during the first 24-48 h of feeding. Of significance, previously validated immunogenic tick saliva proteins including AV422 protein, calreticulin, histamine release factor, histamine binding/lipocalins, selenoproteins, and paramyosin were identified in this screen, supporting the specificity of the approach in this study. While descriptive, this study opens opportunities for in-depth tick feeding physiology studies.
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Proteínas de Artrópodos/genética , Proteoma/metabolismo , Saliva/metabolismo , Garrapatas/metabolismo , Animales , Antígenos/genética , Antígenos/inmunología , Antígenos/metabolismo , Proteínas de Artrópodos/inmunología , Proteínas de Artrópodos/metabolismo , Femenino , Secuenciación de Nucleótidos de Alto Rendimiento , Masculino , Periodo Posprandial , Proteoma/genética , Proteoma/inmunología , Conejos , Saliva/inmunología , Análisis de Secuencia de ADN , Garrapatas/genética , Garrapatas/inmunologíaRESUMEN
This study demonstrates that Amblyomma americanum (Aam) constitutively and ubiquitously expresses the long (L) and short (S) putative acidic chitinases (Ach) that are distinguished by a 210 base pair (bp) deletion in AamAch-S. Full-length AamAch-L and AamAch-S cDNA are 1959 and 1718 bp long, containing 1332 and 1104 bp open reading frames that code for 443 and 367 amino acid residues proteins with the former predicted to be extracellular and the latter intracellular. Both AamAch-L and AamAch-S mRNA are expressed in multiple organs as revealed by qualitative RT-PCR analysis. Furthermore, quantitative reverse transcription polymerase chain reaction analysis revealed that AamAch-L mRNA was downregulated in the mid-gut, but was unchanged in the salivary gland and in other organs in response to feeding. Of significant interest, AamAch-L and/or AamAch-S functions are probably associated with formation and/or maintenance of stability of A. americanum tick cement cone. Dual RNA interference silencing of AamAch-L and/or AamAch-S mRNA caused ticks to loosely attach onto host skin as suggested by bleeding around tick mouthparts and ticks detaching off host skin with a light touch. AamAch-L may apparently encode an inactive chitinase as indicated by Pichia pastoris-expressed recombinant AamAch-L failing to hydrolyse chitinase substrates. Unpublished related work in our laboratory, and published work by others that found AamAch-L in tick saliva, suggest that native AamAch-L is a non-specific immunoglobulin binding tick saliva protein in that rAamAch-L non-specifically bound rabbit, bovine and chicken non-immune sera. We discuss findings in this study with reference to advancing knowledge on tick feeding physiology.
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Quitinasas/fisiología , Conducta Alimentaria/fisiología , Silenciador del Gen , Ixodidae/fisiología , Secuencia de Aminoácidos , Animales , Bovinos , Pollos , Quitinasas/genética , Quitinasas/metabolismo , ADN Complementario/genética , Regulación de la Expresión Génica , Ixodidae/enzimología , Ixodidae/genética , Datos de Secuencia Molecular , Filogenia , Interferencia de ARN , ARN Mensajero/genética , Conejos/parasitología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ProteínaRESUMEN
We present a novel phenotypic readout using inducible, biosynthetic nanoscaffolds to directly visualize dynamic molecular interactions within living cells at the single-cell level with high sensitivity and selectivity. Labeled ferritin is used to form biological nanoparticles inside cells. Specific supramolecular assembly of ferritin-derived nanoparticles induces highly clustered nanoscaffolds. These inducible biosynthetic nanoscaffolds are used as the artificial recruitment/redistribution platform for monitoring interactions of a small molecule with its target protein(s) inside living cells.
Asunto(s)
Nanopartículas , Andamios del Tejido , Ferritinas/química , Colorantes Fluorescentes , Células HeLa , HumanosRESUMEN
BACKGROUND: The development of an efficient method to improve the wound healing process is urgently required for diabetic patients suffering a threat of limb amputations. Various growth factors have been proposed for treatment; however, more research still has to be carried out to maintain their curative effect. In the present study, we describe a simple nonviral gene therapy method for improving wound healing. METHODS: Minicircle plasmid DNA encoding vascular endothelial growth factor (VEGF) was combined with an arginine-grafted cationic dendrimer, PAM-RG4. The formed complexes were injected subcutaneously into the skin wounds of diabetic mice. RESULTS: Actively proliferating cells in wound tissue were efficiently transfected, resulting in a high level of VEGF expression. Within 6 days after injection, skin wounds in the diabetic mice were generally healed and displayed a well-ordered dermal structure, which was confirmed by histological staining. CONCLUSIONS: This simple and effective gene therapy method may represent a powerful tool for the treatment of diabetic foot ulcers and other diseases that are refractory to treatment.
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ADN Circular/administración & dosificación , Dendrímeros/química , Complicaciones de la Diabetes/terapia , Piel/patología , Factor A de Crecimiento Endotelial Vascular/genética , Cicatrización de Heridas , Animales , Arginina/química , Cationes , Complicaciones de la Diabetes/patología , Terapia Genética , Masculino , Ratones , Ratones Endogámicos C57BL , Neovascularización Fisiológica , Piel/irrigación sanguínea , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
Hypertension is the most common comorbidity and major risk factor in patients with erectile dysfunction. The pharmacokinetics of mirodenafil, used for the treatment of erectile dysfunction, after the intravenous and oral administration (20 mg/kg) to 6-week-old rats (with blood pressure within the normotensive range) and 16-week-old spontaneously hypertensive rats (SHRs) and their age-matched control normotensive Kyoto-Wistar (KW) rats, and 16-week-old deoxycorticosterone acetate-salt-induced hypertensive rats (DOCA-salt rats) and their age-matched control Sprague-Dawley (SD) rats were compared. It was found that time-averaged renal clearance (Cl(r)) was of minor importance and that time-averaged non-renal clearance (Cl(nr)) was dominant. In both 6- and 16-week-old SHRs, the Cl(nr)s and areas under the curve (AUCs) of intravenous mirodenafil were significantly smaller and greater than those of the controls, but in 16-week-old DOCA-salt rats, they were comparable to the controls. Although the AUC of oral mirodenafil in 16-week-old SHRs was comparable to the controls, the Cl(nr)s (or total body clearances, Cls) of intravenous mirodenafil and intestinal intrinsic clearances were significantly smaller than the controls and comparable to the controls for both 6- and 16-week-old SHRs, unlike in the 16-week-old DOCA-salt rats. The above data suggest that the significantly smaller Cl(nr) and greater AUC of intravenous mirodenafil and comparable AUC of oral mirodenafil in 16-week-old SHR could be due to the hereditary characteristics of SHRs, and not due to the hypertensive state itself.
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Desoxicorticosterona/toxicidad , Hipertensión/metabolismo , Pirimidinonas/farmacocinética , Sulfonamidas/farmacocinética , Animales , Modelos Animales de Enfermedad , Hipertensión/inducido químicamente , Masculino , Pirimidinonas/metabolismo , Distribución Aleatoria , Ratas , Ratas Endogámicas SHR , Ratas Endogámicas WKY , Ratas Sprague-Dawley , Sulfonamidas/metabolismoRESUMEN
BACKGROUND: The use of second generation supraglottic airway devices is recommended for airway rescue in failed tracheal intubation. This study was performed to compare the clinical performance of the i-gel™ with that of the AuraGain™ in patients with simulated cervical immobilization. METHODS: We conducted a prospective, randomized controlled trial in 104 patients undergoing general anesthesia from June to September 2018 at the Seoul National University Hospital. Patients were randomly allocated to receive either the i-gel™ or the AuraGain™ device. A difficult airway was simulated using a cervical collar limiting the mouth opening and neck movement. The primary outcome was the initial oropharyngeal leak pressure. RESULTS: The rate of successful insertion at the first attempt was 92.3% for the i-gel™ and 86% for the AuraGain™. There were no significant differences in the initial and 5-min oropharyngeal leak pressures between the i-gel™ and the AuraGain™ (21±4 vs. 22±5 cmH2O, P=0.229; and 22±5 vs. 23±5 cmH2O, P=0.308, respectively). The time to successful device insertion was shorter (20 [16-23] vs. 25 [20-41] s, P<0.001) and device insertion was easier (P<0.001) in the i-gel™ group than in the AuraGain™ group. The blood staining of the device was more frequently observed in the AuraGain™ (3 [5.8%] vs. 12 [23.5%] patients, P=0.003). CONCLUSIONS: The i-gel™ and the AuraGain™ showed comparable oropharyngeal leak pressures and success rates in the first attempt in patients with simulated cervical immobilization. However, the i-gel™ was easier to insert and required less time for insertion than the AuraGain™.
Asunto(s)
Máscaras Laríngeas , Anestesia General , Humanos , Intubación Intratraqueal , Cuello , Estudios ProspectivosRESUMEN
BACKGROUND: A novel ultrasound imaging technique, dynamic needle tip positioning (DNTP), enables continuous visualization of the needle tip during ultrasound-guided cannulation. The purpose of this study was to compare the rate of successful first-attempt radial artery cannulations between DNTP and the conventional long-axis in-plane (LAX-IP) technique. METHODS: Adult patients undergoing cardiac surgery requiring radial artery cannulation were included and randomly allocated into either a DNTP or LAX-IP group. Radial artery cannulation was performed by a single experienced practitioner. The primary outcome was the first-attempt success rate of radial artery cannulation. Secondary outcomes included the length of time needed for cannulation and overall incidence of complications. RESULTS: A total of 136 patients were studied. The first-attempt success rate of cannulation was 94% in the DNTP group (N.=70) and 68% in the LAX-IP group (N.=66; OR 7.70, 95% CI 2.48-24.94, P<0.001). The total procedure time was also shorter in the DNTP group (median [IQR]; 87 [72-108] seconds) versus the LAX-IP group (118 [93-182] seconds; P<0.001). During cannulation, vasospasm occurred less often in the DNTP (4%) group compared to the LAX-IP group (17%; P=0.018). CONCLUSIONS: The DNTP technique had a greater first-attempt success rate of radial artery cannulation compared to the conventional LAX-IP technique. Also, DNTP required significantly less time for cannulation and had fewer complications.
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Procedimientos Quirúrgicos Cardíacos/métodos , Cateterismo Periférico/métodos , Arteria Radial/diagnóstico por imagen , Anciano , Cateterismo Periférico/efectos adversos , Femenino , Humanos , Complicaciones Intraoperatorias/epidemiología , Masculino , Persona de Mediana Edad , Agujas , Ultrasonografía IntervencionalRESUMEN
PURPOSE: The purpose of our study was to provide a guide for identification and contouring of upper abdominal organs-at-risk (OARs) in the setting of online magnetic resonance imaging (MRI)-guided radiation treatment planning and delivery. METHODS AND MATERIALS: After a needs assessment survey, it was determined that an upper abdominal MRI-based atlas of normal OARs would be of benefit to radiation oncologists and radiation therapists. An anonymized diagnostic 1.5T MRI from a patient with typical upper abdominal anatomy was used for atlas development. Two MRI sequences were selected for contouring, a T1-weighted gadoxetic acid contrast-enhanced MRI acquired in the hepatobiliary phase and axial fast imaging with balanced steady-state precession. Two additional clinical MRI sequences from commercial online MRI-guided radiation therapy systems were selected for contouring and were included in the final atlas. Contours from each data set were completed and reviewed by radiation oncologists, along with a radiologist who specializes in upper abdominal imaging, to generate a consensus upper abdominal MRI-based OAR atlas. RESULTS: A normal OAR atlas was developed, including recommendations for contouring. The atlas and contouring guidance are described, and high-resolution MRI images and contours are displayed. OARs, such as the bile duct and biliary tree, which may be better seen on MRI than on computed tomography, are highlighted. The full DICOM/DICOM-RT MRI images from both the diagnostic and clinical online MRI-guided radiation therapy systems data sets have been made freely available, for educational purposes, at econtour.org. CONCLUSIONS: This MRI contouring atlas for upper abdominal OARs should provide a useful reference for contouring and education. Its routine use may help to improve uniformity in contouring in radiation oncology planning and OAR dose calculation. Full DICOM/DICOM-RT images are available online and provide a valuable educational resource for upper abdominal MRI-based radiation therapy planning and delivery.
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Abdomen/diagnóstico por imagen , Abdomen/efectos de la radiación , Imagen por Resonancia Magnética , Órganos en Riesgo/diagnóstico por imagen , Órganos en Riesgo/efectos de la radiación , Oncología por Radiación/normas , Humanos , Planificación de la Radioterapia Asistida por Computador , Estándares de ReferenciaRESUMEN
The pharmacokinetics of mirodenafil and its two metabolites, SK3541 and SK3544, after intravenous (5, 10, 20 and 50 mg/kg) and oral (10, 20 and 50 mg/kg) administration of mirodenafil, and the first-pass effect of mirodenafil after intravenous, oral, intraportal, intragastric and intraduodenal (20 mg/kg) administration of mirodenafil were evaluated in rats. The pharmacokinetics of mirodenafil and SK3541 were dose-dependent after both intravenous and oral administration of mirodenafil due to the saturable hepatic metabolism of mirodenafil. After oral administration of mirodenafil, approximately 2.59% of the oral dose was not absorbed, the F value was approximately 29.4%, and the hepatic and gastrointestinal first-pass effects of mirodenafil were approximately 21.4% and 54.3% of the oral dose, respectively. The low F value of mirodenafil in rats was mainly due to considerable hepatic and gastrointestinal first-pass effects in rats. The equilibrium plasma-to-blood cell partition ratios of mirodenafil were independent of the initial blood mirodenafil concentrations of 1-10 microg/ml; the mean values were 1.08-1.21. The plasma binding values of mirodenafil to rat plasma was 87.8%.
Asunto(s)
Erección Peniana/fisiología , Inhibidores de Fosfodiesterasa 5 , Pirimidinonas/administración & dosificación , Pirimidinonas/farmacocinética , Sulfonamidas/administración & dosificación , Sulfonamidas/farmacocinética , Animales , Relación Dosis-Respuesta a Droga , Masculino , Erección Peniana/efectos de los fármacos , Pirimidinonas/química , Ratas , Ratas Sprague-Dawley , Sulfonamidas/química , Distribución Tisular/efectos de los fármacos , Distribución Tisular/fisiologíaRESUMEN
BACKGROUND: Early detection of a risk of postoperative bleeding is essential in cardiac surgery patients. The aim of the present study was to evaluate the utility of the first derivative curve (the V-curve) of the clotting waveform of rotational thromboelastometry (ROTEM) in terms of predicting bleeding after cardiac surgery. METHODS: We retrospectively analysed 534 cardiac surgery patients. We used the chest tube output during the stay in the intensive care unit to divide patients into a higher blood loss group (HBL group; the fourth quartile) and a lower blood loss group (LBL group; the lower quartiles). We performed multivariable logistic regression using the V-curve parameters and potential confounders including conventional ROTEM parameters. RESULTS: In the multivariable model, the adjusted odds ratios for HBL of patients with a lower maximum clotting velocity (MaxVel ≤9 mm*100 s-1) as revealed by extrinsically activated ROTEM (EXTEM), and the area under the velocity curve (AUC ≤988 mm*100) of the fibrin-based extrinsically activated ROTEM (FIBTEM), both measured at skin closure in the end of surgery, were 1.78 (95% CI 1.03 to 3.07) and 2.14 (95% CI 1.20 to 3.82), respectively. However, conventional ROTEM parameters were not included in the final model. Additionally, lower EXTEM MaxVel and FIBTEM AUC values were associated with the need for a higher transfusion volume, longer postoperative intensive care unit and hospital stays, and more frequent re-exploration to control bleeding. CONCLUSIONS: The ROTEM V-curve parameters can predict postoperative bleeding and clinical outcomes after cardiac surgery.
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Procedimientos Quirúrgicos Cardíacos/efectos adversos , Hemorragia Posoperatoria/diagnóstico , Tromboelastografía/métodos , Anciano , Coagulación Sanguínea , Transfusión Sanguínea/estadística & datos numéricos , Cuidados Críticos , Femenino , Humanos , Tiempo de Internación , Masculino , Persona de Mediana Edad , Cuidados Posoperatorios/estadística & datos numéricos , Valor Predictivo de las Pruebas , Estudios Retrospectivos , Análisis de Ondículas , Tiempo de Coagulación de la Sangre TotalRESUMEN
Alzheimer's disease is a neurodegenerative disease characterized by the impairment of cognitive function and loss of memory, affecting millions of individuals worldwide. With the dramatic increase in the prevalence of Alzheimer's disease, it is expected to impose extensive public health and economic burden. However, this burden is particularly heavy on the caregivers of Alzheimer's disease patients eliciting neuropsychiatric symptoms that include mood swings, hallucinations, and depression. Interestingly, these neuropsychiatric symptoms are shared across symptoms of bipolar disorder, schizophrenia, and major depression disorder. Despite the similarities in symptomatology, comorbidities of Alzheimer's disease and these neuropsychiatric disorders have not been studied in the Alzheimer's disease model. Here, we explore the comprehensive changes in gene expression of genes that are associated with bipolar disorder, schizophrenia, and major depression disorder through the microarray of an Alzheimer's disease animal model, the forebrain specific PSEN double knockout mouse. To analyze the genes related with these three neuropsychiatric disorders within the scope of our microarray data, we used selected 1207 of a total of 45,037 genes that satisfied our selection criteria. These genes were selected on the basis of 14 Gene Ontology terms significantly relevant with the three disorders which were identified by previous research conducted by the Psychiatric Genomics Consortium. Our study revealed that the forebrain specific deletion of Alzheimer's disease genes can significantly alter neuropsychiatric disorder associated genes. Most importantly, most of these significantly altered genes were found to be involved with schizophrenia. Taken together, we suggest that the synaptic dysfunction by mutation of Alzheimer's disease genes can lead to the manifestation of not only memory loss and impairments in cognition, but also neuropsychiatric symptoms.
RESUMEN
Understanding the molecular basis of how ticks adapt to feed on different animal hosts is central to understanding tick and tick-borne disease (TBD) epidemiology. There is evidence that ticks differentially express specific sets of genes when stimulated to start feeding. This study was initiated to investigate if ticks such as Ixodes scapularis and Amblyomma americanum that are adapted to feed on multiple hosts utilized the same sets of proteins to prepare for feeding. We exposed I. scapularis and A. americanum to feeding stimuli of different hosts (rabbit, human, and dog) by keeping unfed adult ticks enclosed in a perforated microfuge in close contact with host skin, but not allowing ticks to attach on host. Our data suggest that ticks of the same species differentially express tick saliva proteins (TSPs) when stimulated to start feeding on different hosts. SDS-PAGE and silver staining analysis revealed unique electrophoretic profiles in saliva of I. scapularis and A. americanum that were stimulated to feed on different hosts: rabbit, human, and dog. LC-MS/MS sequencing and pairwise analysis demonstrated that I. scapularis and A. americanum ticks expressed unique protein profiles in their saliva when stimulated to start feeding on different hosts: rabbit, dog, or human. Specifically, our data revealed TSPs that were unique to each treatment and those that were shared between treatments. Overall, we identified a total of 276 and 340 non-redundant I. scapularis and A. americanum TSPs, which we have classified into 28 functional classes including: secreted conserved proteins (unknown functions), proteinase inhibitors, lipocalins, extracellular matrix/cell adhesion, heme/iron metabolism, signal transduction and immunity-related proteins being the most predominant in saliva of unfed ticks. With exception of research on vaccines against Rhipicephalus microplus, which its natural host, cattle, research on vaccine against other ticks relies feeding ticks on laboratory animals. Data here suggest that relying on lab animal tick feeding data to select target antigens could result in prioritizing irrelevant anti-tick vaccine targets that are expressed when ticks feed on laboratory animals. This study provides the platform that could be utilized to identify relevant target anti-tick vaccine antigens, and will facilitate early stage tick feeding research.
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Proteínas de Artrópodos/análisis , Especificidad del Huésped , Ixodidae/fisiología , Saliva/química , Proteínas y Péptidos Salivales/análisis , Animales , Cromatografía Liquida , Perros , Electroforesis en Gel de Poliacrilamida , Humanos , Proteoma/análisis , Conejos , Espectrometría de Masas en TándemRESUMEN
Amblyomma americanum tick serine protease inhibitor (serpin, AAS) 19, is a highly conserved protein that is characterized by its functional domain being 100% conserved across tick species. We also reported that AAS19 was an immunogenic tick saliva protein with anti-haemostatic functions and an inhibitor of trypsin-like proteases including five of the eight serine protease factors in the blood clotting cascade. In this study the goal was to validate the importance of AAS19 in A. americanum tick physiology, assess immunogenicity and investigate tick vaccine efficacy of yeast-expressed recombinant (r) AAS19. We confirm that AAS19 is important to A. americanum fitness and blood meal feeding. AAS19 mRNA disruption by RNAi silencing caused ticks to obtain blood meals that were 50% smaller than controls, and treated ticks being morphologically deformed with 100% of the deformed ticks dying in incubation. We show that rAAS19 is highly immunogenic in that two 500 µg inoculations mixed with TiterMax Gold adjuvant provoked antibody titers of more than 1:320,000 that specifically reacted with native AAS19 in unfed and partially fed tick tissue. Since AAS19 is injected into animals during tick feeding, we challenge infested immunized rabbits twice to test if tick infestations of immunized rabbits could act as booster. While in the first infestation significantly smaller tick blood meals were observed on one of the two immunized rabbits, smaller blood meals were observed on both rabbits, but 60% of ticks that engorged on immunized rabbits in the second infestation failed to lay eggs. It is notable that ticks fed faster on immunized animals despite obtaining smaller blood meals. We conclude that rAAS19 is a potential component of cocktail tick vaccine.
Asunto(s)
Anticuerpos Neutralizantes/biosíntesis , Proteínas de Artrópodos/inmunología , Inhibidores de Serina Proteinasa/inmunología , Infestaciones por Garrapatas/prevención & control , Garrapatas/inmunología , Vacunas Acelulares/inmunología , Vacunas Sintéticas/inmunología , Adyuvantes Inmunológicos/administración & dosificación , Animales , Anticuerpos Neutralizantes/sangre , Proteínas de Artrópodos/administración & dosificación , Proteínas de Artrópodos/genética , Femenino , Expresión Génica , Inmunogenicidad Vacunal , Recuento de Huevos de Parásitos , Pichia/genética , Pichia/metabolismo , Poloxaleno/administración & dosificación , ARN Mensajero/antagonistas & inhibidores , ARN Mensajero/genética , ARN Mensajero/inmunología , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Conejos , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , Saliva/química , Saliva/inmunología , Inhibidores de Serina Proteinasa/administración & dosificación , Inhibidores de Serina Proteinasa/genética , Infestaciones por Garrapatas/inmunología , Infestaciones por Garrapatas/parasitología , Vacunas Acelulares/administración & dosificación , Vacunas Acelulares/genética , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/genéticaRESUMEN
Ixodes scapularis is arguably the most medically important tick species in the United States. This tick transmits 5 of the 14 human tick-borne disease (TBD) agents in the USA: Borrelia burgdorferi, Anaplasma phagocytophilum, B. miyamotoi, Babesia microti, and Powassan virus disease. Except for the Powassan virus disease, I. scapularis-vectored TBD agents require more than 24h post attachment to be transmitted. This study describes identification of 24h immunogenic I. scapularis tick saliva proteins, which could provide opportunities to develop strategies to stop tick feeding before transmission of the majority of pathogens. A 24h fed female I. scapularis phage display cDNA expression library was biopanned using rabbit antibodies to 24h fed I. scapularis female tick saliva proteins, subjected to next generation sequencing, de novo assembly, and bioinformatic analyses. A total of 182 contigs were assembled, of which â¼19% (35/182) are novel and did not show identity to any known proteins in GenBank. The remaining â¼81% (147/182) of contigs were provisionally identified based on matches in GenBank including â¼18% (27/147) that matched protein sequences previously annotated as hypothetical and putative tick saliva proteins. Others include proteases and protease inhibitors (â¼3%, 5/147), transporters and/or ligand binding proteins (â¼6%, 9/147), immunogenic tick saliva housekeeping enzyme-like (17%, 25/147), ribosomal protein-like (â¼31%, 46/147), and those classified as miscellaneous (â¼24%, 35/147). Notable among the miscellaneous class include antimicrobial peptides (microplusin and ricinusin), myosin-like proteins that have been previously found in tick saliva, and heat shock tick saliva protein. Data in this study provides the foundation for in-depth analysis of I. scapularis feeding during the first 24h, before the majority of TBD agents can be transmitted.