RESUMEN

The partially purified water extract from Euglena gracilis (EWE) was evaluated for its antitumor and immunomodulatory effects in cell cultures and in a mouse orthotopic lung carcinoma allograft model. In two-dimensional cell culture, the EWE treatment inhibited cell growth of both murine Lewis lung carcinoma (LLC) and human lung carcinoma cells (A549 and H1299) in a dose- and time-dependent manner. In contrast, the growth of mouse bone marrow cells (BMCs), but not mouse splenocytes (SPLs), was stimulated by the treatment with EWE. In three-dimensional spheroid culture, spheroid growth of LLC cells was significantly attenuated by EWE treatment. In a mouse LLC orthotopic allograft model, pretreatment with EWE (150-200 mg/kg/day, via drinking water) three weeks prior to the LLC cell inoculation, but not post-treatment after LLC cell inoculation, significantly attenuated the growth of LLC tumors in immunocompetent syngeneic mouse lung. This tumor growth attenuation coincided with a significant decrease in the population of myeloid-derived cells, primarily neutrophils. Flow cytometric analysis revealed that the EWE treatment significantly attenuated growth of granulocytic myeloid-derived suppressor cells (gMDSC) in BMCs and that this decrease was due to induction of gMDSC-specific apoptosis and differentiation of monocytic MDSCs (mMDSC) to macrophages. The present study provides evidence that EWE pretreatment inhibits lung carcinoma growth mainly by stimulating host antitumor immunity through attenuation of growth of gMDSCs and decreasing the number of peripheral granulocytes. This study suggests that the partially purified extract derived from Euglena gracilis contains significant bioactive materials that prevent lung carcinoma growth.

Asunto(s)

Antineoplásicos/farmacología , Carcinoma Pulmonar de Lewis/tratamiento farmacológico , Euglena gracilis/metabolismo , Neoplasias Pulmonares/tratamiento farmacológico , Células A549 , Animales , Antineoplásicos/administración & dosificación , Antineoplásicos/aislamiento & purificación , Apoptosis/efectos de los fármacos , Células de la Médula Ósea/efectos de los fármacos , Células de la Médula Ósea/metabolismo , Carcinoma Pulmonar de Lewis/patología , Diferenciación Celular/efectos de los fármacos , Línea Celular Tumoral , Relación Dosis-Respuesta a Droga , Femenino , Granulocitos/efectos de los fármacos , Granulocitos/metabolismo , Humanos , Factores Inmunológicos/administración & dosificación , Factores Inmunológicos/aislamiento & purificación , Factores Inmunológicos/farmacología , Neoplasias Pulmonares/patología , Ratones , Ratones Endogámicos C57BL , Células Supresoras de Origen Mieloide/metabolismo , Factores de Tiempo , Agua/química

RESUMEN

OBJECTIVES: To describe current trends in filled opioid prescriptions for Medicaid-enrolled children, adolescents and young adults (AYAs) from 2012 to 2016, and to identify patient characteristics and clinical settings associated with a higher probability of filled opioid prescriptions. DESIGN: Retrospective cohort study of children and young adults enrolled in Medicaid from 2012 to 2016. SETTING: 10-12 states participating in the Medicaid Marketscan claims database. PARTICIPANTS: Medicaid-enrolled children and young adults (0-21 years old). EXPOSURE: Healthcare encounter(s) that could result in a new opioid prescription. MAIN OUTCOME MEASURE: "Opioid visits," defined as healthcare encounters associated with a new opioid prescription filled within 7 days. Each opioid visit was assigned to the clinical provider most likely to have prescribed an opioid. RESULTS: There were 113,068,027 visits among 4,427,838 Medicaid-enrollees and 1 percent (n = 1,130,006) of these were considered an opioid visit. Adjusted probabilities decreased from 1.2 percent to 0.8 percent from 2012 to 2016. The most frequently prescribed opioids were hydrocodone (48 percent; n = 653,011), codeine (23 percent; n = 305,644), and oxycodone (14 percent; n = 189,700); most of these were in combination with acetaminophen. The high-est adjusted percentages by clinical setting were seen in dental surgery (29 percent), outpatient surgery (21 percent), and inpatient (upon discharge, 10 percent). CONCLUSIONS: Opioid prescriptions filled for Medicaid-enrolled children, adolescents, and young adults are relatively rare and adjusted probabilities decreased from 2012 to 2016. Among opioids filled, combination opioids and those with pedi-atric safety warnings remain commonly prescribed. Further research is critical to better understand drivers of prescribing practices and clinical indications for appropriate opioid use to inform improvements in pain management guidelines in this population.

Asunto(s)

Analgésicos Opioides/administración & dosificación , Prescripciones de Medicamentos/estadística & datos numéricos , Medicaid , Pautas de la Práctica en Medicina/tendencias , Adolescente , Niño , Preescolar , Humanos , Lactante , Recién Nacido , Estudios Retrospectivos , Estados Unidos/epidemiología , Adulto Joven

RESUMEN

Fitness is determined by the ability of an organism to both survive and reproduce; however, the mechanisms that lead to increased survival may not have the same effect on reproductive success. We used nineteen natural Drosophila melanogaster genotypes from the Drosophila Genetic Reference Panel to determine if adaptive plasticity following short-term acclimation through rapid cold-hardening (RCH) affects mating behavior and mating success. We confirmed that exposure to the acclimation temperature is beneficial to survival following cold stress; however, we found that this same acclimation temperature exposure led to less efficient male courtship and a significant decrease in the likelihood of mating. Cold tolerance and the capacity to respond plastically to cold stress were not correlated with mating behavior following acclimation, suggesting that the genetic control of the physiological effects of the cold temperature exposure likely differ between survival and behavioral responses. We also tested whether the exposure of males to the acclimation temperature influenced courtship song. This exposure again significantly increased courtship duration; however, courtship song was unchanged. These results illustrate costs of short-term acclimation on survival and reproductive components of fitness and demonstrate the pronounced effect that short-term thermal environment shifts can have on reproductive success.

Asunto(s)

Aclimatación , Frío , Drosophila melanogaster/genética , Drosophila melanogaster/fisiología , Genotipo , Conducta Sexual Animal/fisiología , Animales , Femenino , Variación Genética , Masculino , Análisis de Supervivencia , Factores de Tiempo , Vocalización Animal/fisiología

RESUMEN

To evaluate the potential of cell-penetrating peptide-based delivery of apoptosis-inducer gene in cancer therapy, a modified HIV-1 TAT peptide (dimerized TAT peptide, dTAT) was studied. The dTAT and plasmid DNA (pDNA) complexes (dTAT-pDNA) were condensed using calcium chloride (dTAT-pDNA-Ca2+). This simple nonviral formulation approach showed high levels of gene expression in vitro without any cytotoxicity. In mouse studies, a single intratracheal (IT) aerosol spray or 2 intravenous (IV) injections of the dTAT, apoptosis-inducer gene, angiotensin II type 2 receptor (AT2R), and Ca2+ complexes (dTAT-pAT2R-Ca2+) significantly attenuated the acutely growing mouse Lewis lung carcinoma allografts in mouse lungs. Furthermore, single IT (p = 0.054) and the combination of IT and IV (p < 0.05) administrations of dTAT-pAT2R-Ca2+ markedly attenuated slowly growing and relatively large-sized H358 human bronchioloalveolar carcinoma xenografts in mouse lungs. These results indicate that the dTAT-pDNA-Ca2+ effectively delivered the gene to cancer cells by either IT or IV administration although the local pulmonary delivery of the dTAT-pAT2R-Ca2+ showed more effective growth inhibition of orthotopic lung cancer grafts. Thus, the present study offers preclinical proof of concept that a dTAT-based nonviral gene delivery method via IT administration may be an effective lung cancer gene therapy.

Asunto(s)

Carcinoma Pulmonar de Lewis/terapia , ADN/administración & dosificación , Técnicas de Transferencia de Gen , Neoplasias Pulmonares/terapia , Nanopartículas/química , Receptor de Angiotensina Tipo 2/genética , Productos del Gen tat del Virus de la Inmunodeficiencia Humana/química , Animales , Carcinoma Pulmonar de Lewis/genética , Línea Celular Tumoral , ADN/genética , ADN/uso terapéutico , Expresión Génica , Terapia Genética , VIH-1/química , Humanos , Pulmón/metabolismo , Pulmón/patología , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones SCID , Plásmidos/administración & dosificación , Plásmidos/genética , Plásmidos/uso terapéutico

RESUMEN

Umbilical cord derived mesenchymal stromal cells (UC-MSCs) are a focus for clinical translation but standardized methods for isolation and expansion are lacking. Previously we published isolation and expansion methods for UC-MSCs which presented challenges when considering good manufacturing practices (GMP) for clinical translation. Here, a new and more standardized method for isolation and expansion of UC-MSCs is described. The new method eliminates dissection of blood vessels and uses a closed-vessel dissociation following enzymatic digestion which reduces contamination risk and manipulation time. The new method produced >10 times more cells per cm of UC than our previous method. When biographical variables were compared, more UC-MSCs per gram were isolated after vaginal birth compared to Caesarian-section births, an unexpected result. UC-MSCs were expanded in medium enriched with 2%, 5%, or 10% pooled human platelet lysate (HPL) eliminating the xenogeneic serum components. When the HPL concentrations were compared, media supplemented with 10% HPL had the highest growth rate, smallest cells, and the most viable cells at passage. UC-MSCs grown in 10% HPL had surface marker expression typical of MSCs, high colony forming efficiency, and could undergo trilineage differentiation. The new protocol standardizes manufacturing of UC-MSCs and enables clinical translation.