Monocyte to macrophage differentiation and changes in cellular redox homeostasis promote cell type-specific HIV latency reactivation.

HIV latency regulation in monocytes and macrophages can vary according to signals directing differentiation, polarization, and function. To investigate these processes, we generated an HIV latency model in THP-1 monocytes and showed differential levels of HIV reactivation among clonal populations. Monocyte-to-macrophage differentiation of HIV-infected primary human CD14+ and THP-1 cells induced HIV reactivation and showed that virus production increased concomitant with macrophage differentiation. We applied the HIV-infected THP-1 monocyte-to-macrophage (MLat) model to assess the biological mechanisms regulating HIV latency dynamics during monocyte-to-macrophage differentiation. We pinpointed protein kinase C signaling pathway activation and Cyclin T1 upregulation as inherent differentiation mechanisms that regulate HIV latency reactivation. Macrophage polarization regulated latency, revealing proinflammatory M1 macrophages suppressed HIV reactivation while anti-inflammatory M2 macrophages promoted HIV reactivation. Because macrophages rely on reactive-oxygen species (ROS) to exert numerous cellular functions, we disrupted redox pathways and found that inhibitors of the thioredoxin (Trx) system acted as latency-promoting agents in T-cells and monocytes, but opposingly acted as latency-reversing agents in macrophages. We explored this mechanism with Auranofin, a clinical candidate for reducing HIV reservoirs, and demonstrated Trx reductase inhibition led to ROS induced NF-κB activity, which promoted HIV reactivation in macrophages, but not in T-cells and monocytes. Collectively, cell type-specific differences in HIV latency regulation could pose a barrier to HIV eradication strategies.

Evaluation of the use of individualized patient care plans in frequent emergency department visitors with pain complaints.

BACKGROUND: Pain is one of the most common complaints that patients present to the emergency department for; emergency medicine providers are tasked with providing appropriate pain relief while simultaneously limiting the risk of personal and societal harm that may result from opioid misuse. The Lakeland Regional Medical Center developed a medical management program that identified frequent emergency department visitors with a chief complaint of pain. Individualized care plans were developed for these patients. A retrospective review was then conducted to assess the efficacy of these care plans in reducing the number of emergency department visits for pain-related complaints by the patients entered into the medical management program. RESULTS: There were 294 patients; 65% were male, and the median age was 41 (interquartile range: 33 to 51). A total of 80% percent of the patients were white, and the payors were as follows: 53% were self-pay, 42% were government programs, and 5% had private insurance. The three most common chronic pain complaints were 39% abdominal pain, 24% back/neck pain, and 23% headache/migraine (patients could have more than one area of pain). A total of 60% of the patients had a primary care provider, and another 18% had a pain management provider in addition to primary care. Post plan admissions were significantly reduced to a median of 1 (IQR 0 to 3) with the Wilcoxon signed-rank test's p-value of less than 0.001. CONCLUSION: The authors describe their experience with a quality improvement initiative that identifies frequent emergency department visitors with a chief complaint of pain and provides individualized care plans to these patients. The goals of the program are to improve patient's quality and consistency of care, through interventions that eliminate the prescribing of opioids while providing non-opioid alternatives.

Disseminated Intravascular Coagulation in Varying Age Groups Based on Clinical Conditions.

Disseminated intravascular coagulation (DIC) is a serious syndrome characterized by the systemic activation of blood coagulation resulting in the thrombosis of vessels leading to organ dysfunction and severe bleeding. When physicians try to treat DIC, it is imperative to diagnose and treat the underlying conditions. Anyone can be affected by DIC, but vulnerable groups such as pediatric populations, pregnant women and the elderly may be at higher risk. In this review, the current literature on DIC in pregnancy, the pediatric population, and the elderly is reported. This review also highlights the similarities and differences in the etiology, clinical presentation, diagnosis, and management of DIC in the aforementioned groups (i.e., pediatrics, pregnant women, and the elderly). Findings from this study may help increase awareness about various presentations of DIC in these groups to facilitate rapid recognition of symptoms leading to correct diagnoses.

Synergistic Chromatin-Modifying Treatments Reactivate Latent HIV and Decrease Migration of Multiple Host-Cell Types.

Upon infection of its host cell, human immunodeficiency virus (HIV) establishes a quiescent and non-productive state capable of spontaneous reactivation. Diverse cell types harboring the provirus form a latent reservoir, constituting a major obstacle to curing HIV. Here, we investigate the effects of latency reversal agents (LRAs) in an HIV-infected THP-1 monocyte cell line in vitro. We demonstrate that leading drug treatments synergize activation of the HIV long terminal repeat (LTR) promoter. We establish a latency model in THP-1 monocytes using a replication incompetent HIV reporter vector with functional Tat, and show that chromatin modifiers synergize with a potent transcriptional activator to enhance HIV reactivation, similar to T-cells. Furthermore, leading reactivation cocktails are shown to differentially affect latency reactivation and surface expression of chemokine receptor type 4 (CXCR4), leading to altered host cell migration. This study investigates the effect of chromatin-modifying LRA treatments on HIV latent reactivation and cell migration in monocytes. As previously reported in T-cells, epigenetic mechanisms in monocytes contribute to controlling the relationship between latent reactivation and cell migration. Ultimately, advanced "Shock and Kill" therapy needs to successfully target and account for all host cell types represented in a complex and composite latency milieu.

Micro-Clotting of Platelet-Rich Plasma Upon Loading in Hydrogel Microspheres Leads to Prolonged Protein Release and Slower Microsphere Degradation.

Platelet-rich plasma (PRP) is an autologous blood product that contains a variety of growth factors (GFs) that are released upon platelet activation. Despite some therapeutic potential of PRP in vitro, in vivo data are not convincing. Bolus injection of PRP is cleared rapidly from the body diminishing its therapeutic efficacy. This highlights a need for a delivery vehicle for a sustained release of PRP to improve its therapeutic effect. In this study, we used microfluidics to fabricate biodegradable PRP-loaded polyethylene glycol (PEG) microspheres. PRP was incorporated into the microspheres as a lyophilized PRP powder either as is (powder PRP) or first solubilized and pre-clotted to remove clots (liquid PRP). A high PRP loading of 10% w/v was achieved for both PRP preparations. We characterized the properties of the resulting PRP-loaded PEG microspheres including swelling, modulus, degradation, and protein release as a function of PRP loading and preparation. Overall, loading powder PRP into the PEG microspheres significantly affected the properties of microspheres, with the most pronounced effect noted in degradation. We further determined that microsphere degradation in the presence of powder PRP was affected by platelet aggregation and clotting. Platelet aggregation did not prevent but prolonged sustained PRP release from the microspheres. The delivery system developed and characterized herein could be useful for the loading and releasing of PRP to promote tissue regeneration and wound healing or to suppress tissue degeneration in osteoarthritis, and intervertebral disc degeneration.

Platelet-Rich Plasma Released From Polyethylene Glycol Hydrogels Exerts Beneficial Effects on Human Chondrocytes.

Osteoarthritis (OA) is a debilitating joint disease resulting from chronic joint inflammation and erosion of articular cartilage. A promising biological treatment for OA is intra-articular administration of platelet-rich plasma (PRP). However, immediate bolus release of growth factors limits beneficial therapeutic effects of PRP, thus necessitating the demand for sustained release platforms. In this study, we evaluated the therapeutic value of PRP released from a polyethylene glycol (PEG) hydrogel on articular chondrocytes/cartilage explants derived from OA patients. Lyophilized PRP (PRGF) was encapsulated in PEG hydrogels at 10% w/v and hydrogel swelling, storage modulus and degradation and PRGF release kinetics were determined. PRGF releasate from the hydrogels was collected on day 1, 4, and 11. Encapsulation of PRGF at 10% w/v in PEG hydrogels had minimal effect on hydrogel properties. PRGF was released with an initial burst followed by sustained release until complete hydrogel degradation. Effect of PRGF releasates and bolus PRGF (1% w/v PRGF) on patient-derived cartilage explants or chondrocytes was assessed by chondrocyte proliferation (pico-green assay), gene expression for COL1A1, COL2A1, MMP13, COX2, and NFKB1 (real-time polymerase chain reaction), and measurement of nitric oxide concentration (Griess' assay). Compared to bolus PRGF, PRGF releasates enhanced chondrocyte proliferation, suppressed the expression of genes like MMP13, NFKB1, COL1A1, and COL2A1 and reduced levels of nitric oxide. Taken together, these results indicate that release of PRGF from PEG hydrogels may improve the therapeutic efficacy of PRP and merits further investigation in an animal model of OA. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:2401-2410, 2019.

Morphological adaptations in breast cancer cells as a function of prolonged passaging on compliant substrates.

Standard tissue culture practices involve propagating cells on tissue culture polystyrene (TCP) dishes, which are flat, 2-dimensional (2D) and orders of magnitude stiffer than most tissues in the body. Such simplified conditions lead to phenotypical cell changes and altered cell behaviors. Hence, much research has been focused on developing novel biomaterials and culture conditions that more closely emulate in vivo cell microenvironments. In particular, biomaterial stiffness has emerged as a key property that greatly affects cell behaviors such as adhesion, morphology, proliferation and motility among others. Here we ask whether cells that have been conditioned to TCP, would still show significant dependence on substrate stiffness if they are first pre-adapted to a more physiologically relevant environment. We used two commonly utilized breast cancer cell lines, namely MDA-MB-231 and MCF-7, and examined the effect of prolonged cell culturing on polyacrylamide substrates of varying compliance. We followed changes in cell adhesion, proliferation, shape factor, spreading area and spreading rate. After pre-adaptation, we noted diminished differences in cell behaviors when comparing between soft (1 kPa) and stiff (103 kPa) gels as well as rigid TCP control. Prolonged culturing of cells on complaint substrates further influenced responses of pre-adapted cells when transferred back to TCP. Our results have implications for the study of stiffness-dependent cell behaviors and indicate that cell pre-adaptation to the substrate needs consideration.

Resistencia a carbapenemas en Pseudomonas aeruginosa aisladas en urocultivos: prevalencia y factores de riesgo / Carbapenem resistance in Pseudomonas aeruginosa isolated from urine cultures: prevalence and risk factors

Introducción. Pseudomonas aeruginosa es un bacilo gramnegativo no fermentador con una gran capacidad para desarrollar resistencia a múltiples antimicrobianos, incluidas las carbapenemas, lo que supone un problema creciente a nivel mundial. El objetivo de este estudio fue analizar la prevalencia de P. aeruginosa resistente a carbapenemas (PARC) en urocultivos y evaluar los factores de riesgo asociados al desarrollo de dicho patrón de resistencia. Material y métodos. Se seleccionaron los urocultivos positivos para P. aeruginosa realizados en nuestro hospital entre septiembre de 2012 y septiembre de 2014. Se excluyeron los cultivos repetidos procedentes del mismo paciente. Se creó una base de datos con diversas variables, incluyendo resistencias antimicrobianas. Se calculó la prevalencia de resistencia a carbapenemas y se analizaron los factores de riesgo para crecimiento de PARC. Resultados. Se incluyeron 91 urocultivos positivos para P. aeruginosa. La prevalencia de PARC fue del 22%. Los factores asociados al crecimiento de PARC en el análisis univariante fueron: insuficiencia cardíaca congestiva (p=0,02), tratamiento previo con ampicilina (p=0,04), meropenem (p=0,04), piperacilina-tazobactam (p=0,01), cotrimoxazol (p=0,01) y tratamiento previo con más de un antibiótico (p<0,01). Solamente la insuficiencia cardíaca congestiva (p<0,01) y el tratamiento previo con más de un antibiótico (p<0,01) mostraron diferencias significativas en el análisis multivariante. Conclusiones. La prevalencia de PARC en urocultivos es elevada en nuestro medio. Debemos considerar la presencia de factores de riesgo como el tratamiento previo con más de un antibiótico o la presencia de comorbilidades como la insuficiencia cardíaca para seleccionar una antibioterapia empírica adecuada en pacientes con infecciones del tracto urinario graves (AU)

Introduction. Pseudomonas aeruginosa is a non-fermentative gram-negative bacillus with a great ability to develop resistance to multiple antibiotics, including carbapenems, which is a growing problem worldwide. The aim of this study was to analyse the prevalence of carbapenem-resistant P. aeruginosa (CRPA) in urine cultures and to determine the risk factors associated with the development of carbapanem resistance. Material and method. Positive urine cultures to P. aeruginosa between September 2012 and September 2014 were identified. We excluded repetitive cultures from the same patient. We created a database with different variables, including antimicrobial resistance. The prevalence of carbapenem resistance and the risk factors for growth of CRPA were analysed. Results. Ninety-one patients with positive urine cultures to P. aeruginosa were included. The prevalence of CRPA was 22%. The risk factors to CRPA infection in the univariate analysis were: congestive heart failure (p=0.02), previous treatment with ampicillin (p=0.04), meropenem (p=0.04), piperacillin-tazobactam (p=0.01), trimethoprim-sulfamethoxazole (p= 0.01) and previous treatment with more than one antibiotic (p<0.01). Only congestive heart failure (p<0.01) and previous treatment with more than one antibiotic (p<0.01) showed statistically significant differences in the multivariate analysis. Conclusions. The prevalence of CRPA in urine cultures is high in our population. We should assess the presence of risk factors as previous treatment with more than one antibiotic or comorbidities such as heart failure, in order to select an appropriate empirical treatment in patients with severe urinary tract infections (AU)

Pseudomonas aeruginosa urinary tract infections in hospitalized patients: Mortality and prognostic factors.

BACKGROUND: The aim of this study was to analyze the mortality and predictors of 30-day mortality among hospitalized patients with Pseudomonas aeruginosa urinary tract infection (PAUTI) and the impact of antibiotic treatment on survival. METHODS: Patients admitted to our hospital with PAUTI or those diagnosed of PAUTI during hospitalization for other disease between September 2012 and September 2014 were included. Repeated episodes from the same patient were excluded. Database with demographic, clinical and laboratory ítems was created. Empirical and definitive antibiotic therapy, antimicrobial resistance and all-cause mortality at 30 and 90 days were included. RESULTS: 62 patients were included, with a mean age of 75 years. 51% were male. Mortality was 17.7% at 30 days and 33.9% at 90 days. Factors associated with reduced survival at 30 days were chronic liver disease with portal hypertension (P<0,01), diabetes mellitus (P = 0,04) chronic renal failure (P = 0,02), severe sepsis or septic shock (P<0,01), Charlson index > 3 (P = 0.02) and inadequated definitive antibiotic treatment (P<0,01). Independent risk factors for mortality in multivariate analysis were advanced chronic liver disease (HR 77,4; P<0,01), diabetes mellitus (HR 3,6; P = 0,04), chronic renal failure (HR 4,1; P = 0,03) and inadequated definitive antimicrobial treatment (HR 6,8; P = 0,01). CONCLUSIONS: PAUTI are associated with high mortality in hospitalized patients, which increases significantly in those with severe comorbidity such as chronic renal failure, advanced liver disease or diabetes mellitus. Inadequated antibiotic treatment is associated with poor outcome, which remarks the importance of adjusting empirical antibiotic treatment based on the microbiological susceptibility results.

La legislación a favor de la adolescente embarazada / Legislation in favor of pregnant adolescent

Introducción: la legislación y las políticas son herramientas necesarias para lograr el desarrollo y la evolución en todos los aspectos de un país. Para el profesional de enfermería la creación de estrategias de cuidado y mejoramiento depende del conocimiento que tenga sobre estos lincamientos y de la formación académica, logrando aportar como ciencia independiente disminución de problemáticas como es el embarazo adolescente. Materiales y Métodos: Mediante una búsqueda de literatura y el análisis de la legislación en Colombia se realiza una reflexión sobre la situación de la adolescente embarazada. Resultados: Se logra evidenciar que en Colombia y el mundo existen herramientas legislativas y estrategias de apoyo a los niños, niñas y adolescentes, y están en búsqueda de disminuir la situación de embarazos a temprana edad. Así mismo se destaca a la Enfermera eje fundamental en la salud de la población como ente educador. Discusión y Conclusiones: Es de suma importancia que la Enfermera(o) no limite la consulta o la atención a la necesidad del momento sino que tenga un alcance más amplio, que tenga una visión de la persona identificando necesidades propias de la edad y apoyada en las políticas y legislación del país realizar un cuidado integral. Se recomienda que haya más flujo de información de los derechos sexuales y reproductivos en toda la población, donde se destaquen aspectos como proyecto de vida y refuerzo de valores.

Introduction: the law and policy are necessary tools for development and evolution in all aspects of a country. For the nurse creation care and improvement strategies depends on the knowledge you have about these guidelines and academic training, managing to provide independent science decline as problematic as teenage pregnancy. Materials and Methods: A search of literature and an analysis of the laws in Colombia as a reflection on the situation of pregnant adolescents. Results: We show that succeeds in Colombia and the world there are legislative tools and strategies to support children and adolescents, and are seeking to diminish the status of early pregnancy. It also highlights the fundamental axis Nurse health of the population as being an educator. Discussion and Conclusions: It is extremely important that the nurse does not limit the query or the attention to the need of the moment but have a broader scope, with a vision of the person identifying needs of the age and supported in policies and legislation in the country to perform a comprehensive care. It is recommended that more flow of information on sexual and reproductive rights entire population, where as a project highlighting aspects of life and reinforce values.