Activation-induced endocytosis of the raft-associated transmembrane adaptor protein LAB/NTAL in B lymphocytes: evidence for a role in internalization of the B cell receptor

Linker for activation of B cell (LAB)/non-T cell activation linker (NTAL) and phosphoprotein associated with glycophospholipid-enriched membrane microdomain (PAG)/Csk-binding protein (Cbp) are raft-associated transmembrane adaptor proteins with distinct functions in immediate/early phases of receptor signaling pathways. Heterogeneous rafts are thought to compartmentalize membrane-associated signaling events. In order to investigate the subcellular localization of LAB/NTAL and PAG/Cbp, they were expressed as fluorescent chimeric fusion proteins in a human B cell line and their distribution was examined, along with the corresponding endogenous proteins, before and after B cell receptor (BCR) stimulation. Both adaptors were distributed predominantly at the plasma membrane in resting cells and co-clustered with other raft-associated proteins; however, they distributed differently in buoyant membranes isolated by either detergent resistance or non-detergent methods, indicating that they might localize to distinct rafts. After activation, LAB/NTAL was internalized and co-localized with the BCR while PAG/Cbp remained on the cell surface. BCR internalization was reduced in LAB/NTAL-deficient murine B cells, suggesting a regulatory role for LAB/NTAL in activation-induced internalization of the BCR. The cytoplasmic domain of LAB/NTAL, and not the transmembrane/juxtamembrane region, was found to be essential for its internalization.     

Marijuana Use and Driving Under the Influence among Young Adults: A Socioecological Perspective on Risk Factors

Background: Given increases in marijuana use and driving under the influence (DUI), it is critical to identify those at risk in order to inform intervention efforts. Objectives: We used a socioecological framework to examine correlates of level of marijuana use and DUI in the past month among young adult marijuana users. Methods: We recruited 1567 participants aged 18–34 years via Facebook ads targeting tobacco and marijuana users in August 2014 to complete an online survey assessing marijuana use and DUI, as well as related multilevel factors. Analyses focused on 649 participants reporting past 30-day marijuana use. Results: The sample was an average age of 24.48 (SD = 5.10), 43.9% female, and 76.4% White and used marijuana an average of 17.86 (SD = 11.29) days in the past month. Notably, 48.4% reported driving after marijuana use at least once in the past month, and 74.0% were passengers. Multivariable regression indicated that greater use was associated with: being older; being male; greater symptoms of dependence; residing in a state with recreational marijuana legalized; having a medical marijuana card; having parents and more friends who use; higher coping motives; lower perceived harm to health; and less concern about driving after marijuana use (adjusted R-squared = 0.294). Correlates of driving after using marijuana in the past month included: being younger; more frequent use; having more friends who use; higher enhancement motives; and less concern about driving after using (Nagelkerke R-squared = 0.442). Conclusions/Importance: Interventions and campaigns should address social norms and risk perceptions regarding marijuana use, particularly as it relates to DUI.     

Hair dryer use to optimize pulsed dye laser treatment in rosacea patients

Rosacea is a common chronic inflammatory condition characterized by erythema, telangiectasias, papules, and pustules. While there are many effective treatment options for the papulopustular type, laser therapy remains the most effective modality to treat erythematotelangiectatic rosacea. Erythema and flushing associated with rosacea remains an uncomfortable and socially embarrassing problem for patients. Unfortunately, patients often do not have significant erythema or flushing when they present for laser treatment. With this in mind, we propose a novel technique aimed at enhancing the response of rosacea patients being treated for erythema with pulsed dye laser. Specifically, we present a split-face example of our clinical observation that pre-treatment with forced heated air prior to pulsed-dye laser leads to a greater response in rosacea patients with erythema and flushing.     

Role of microRNA-23b in flow-regulation of Rb phosphorylation and endothelial cell growth

MicroRNAs (miRs) can regulate many cellular functions, but their roles in regulating responses of vascular endothelial cells (ECs) to mechanical stimuli remain unexplored. We hypothesize that the physiological responses of ECs are regulated by not only mRNA and protein signaling networks, but also expression of the corresponding miRs. EC growth arrest induced by pulsatile shear (PS) flow is an important feature for flow regulation of ECs. miR profiling showed that 21 miRs are differentially expressed (8 up- and 13 downregulated) in response to 24-h PS as compared to static condition (ST). The mRNA expression profile indicates EC growth arrest under 24-h PS. Analysis of differentially expressed miRs yielded 68 predicted mRNA targets that overlapped with results of microarray mRNA profiling. Functional analysis of miR profile indicates that the cell cycle network is highly regulated. The upregulation of miR-23b and miR-27b was found to correlate with the PS-induced EC growth arrest. Inhibition of miR-23b using antagomir-23b oligonucleotide (AM23b) reversed the PS-induced E2F1 reduction and retinoblastoma (Rb) hypophosphorylation and attenuated the PS-induced G1/G0 arrest. Antagomir AM27b regulated E2F1 expression, but did not affect Rb and growth arrest. Our findings indicate that PS suppresses EC proliferation through the regulation of miR-23b and provide insights into the role of miRs in mechanotransduction.     

Cystic fibrosis presenting as bilateral maxillary mucoceles in an infant

A six months old male presented to our Otolaryngology Department with a long -standing history of chronic nasal congestion and difficulty of breathing. The child was initially treated with nasal steroids without improvements. On follow up investigations, bilateral sinus mucoceles were seen on CT scan. The diagnosis of cystic fibrosis was confirmed by the chloride sweat test and the child was taken to the operating room for functional endoscopic sinus surgery and marsupialization of mucoceles. The child's brother was then investigated and diagnosed with CF.     

Hydrophobic Gentamicin-Loaded Nanoparticles Are Effective against Brucella melitensis Infection in Mice

The clinical management of human brucellosis is still challenging and demands in vitro active antibiotics capable of targeting the pathogen-harboring intracellular compartments. A sustained release of the antibiotic at the site of infection would make it possible to reduce the number of required doses and thus the treatment-associated toxicity. In this study, a hydrophobically modified gentamicin, gentamicin-AOT [AOT is bis(2-ethylhexyl) sulfosuccinate sodium salt], was either microstructured or encapsulated in poly(lactic-co-glycolic acid) (PLGA) nanoparticles. The efficacy of the formulations developed was studied both in vitro and in vivo. Gentamicin formulations reduced Brucella infection in experimentally infected THP-1 monocytes (>2-log₁₀ unit reduction) when using clinically relevant concentrations (18 mg/liter). Moreover, in vivo studies demonstrated that gentamicin-AOT-loaded nanoparticles efficiently targeted the drug both to the liver and the spleen and maintained an antibiotic therapeutic concentration for up to 4 days in both organs. This resulted in an improved efficacy of the antibiotic in experimentally infected mice. Thus, while 14 doses of free gentamicin did not alter the course of the infection, only 4 doses of gentamicin-AOT-loaded nanoparticles reduced the splenic infection by 3.23 logs and eliminated it from 50% of the infected mice with no evidence of adverse toxic effects. These results strongly suggest that PLGA nanoparticles containing chemically modified hydrophobic gentamicin may be a promising alternative for the treatment of human brucellosis.     

Perinatal and early postnatal reorganization of the subplate and related cellular compartments in the human cerebral wall as revealed by histological and MRI approaches

We analyzed the developmental history of the subplate and related cellular compartments of the prenatal and early postnatal human cerebrum by combining postmortem histological analysis with in vivo MRI. Histological analysis was performed on 21 postmortem brains (age range: 26 postconceptional weeks to 6.5 years) using Nissl staining, AChE-histochemistry, PAS–Alcian blue histochemistry, Gallyas’ silver impregnation, and immunocytochemistry for MAP2, synaptophysin, neurofilament, chondroitin sulfate, fibronectin, and myelin basic protein. The histological findings were correlated with in vivo MRI findings obtained in 30 age-matched fetuses, infants, and children. We analyzed developmental reorganization of major cellular (cell bodies, growing axons) and extracellular (extracellular matrix) components of the subplate and the developing cortex/white matter interface. We found that perinatal and postnatal reorganization of these tissue components is protracted (extending into the second year of life) and characterized by well-delineated, transient and previously undescribed structural and molecular changes at the cortex/white matter interface. The findings of this study are clinically relevant because they may inform and guide a proper interpretation of highly dynamic and hitherto puzzling changes of cortical thickness and cortical/white matter interface as described in current in vivo MRI studies.