The pan-histone deacetylase inhibitor panobinostat is a potential therapy for malignant glioma, but it is water insoluble and does not cross the blood-brain barrier when administered systemically. In ...this article, we describe the in vitro and in vivo efficacy of a novel water-soluble nano-micellar formulation of panobinostat designed for administration by convection enhanced delivery (CED).
The in vitro efficacy of panobinostat-loaded nano-micelles against rat F98, human U87-MG and M059K glioma cells and against patient-derived glioma stem cells was measured using a cell viability assay. Nano-micelle distribution in rat brain was analyzed following acute CED using rhodamine-labeled nano-micelles, and toxicity was assayed using immunofluorescent microscopy and synaptophysin enzyme-linked immunosorbent assay. We compared the survival of the bioluminescent syngenic F98/Fischer344 rat glioblastoma model treated by acute CED of panobinostat-loaded nano-micelles with that of untreated and vehicle-only-treated controls.
Nano-micellar panobinostat is cytotoxic to rat and human glioma cells in vitro in a dose-dependent manner following short-time exposure to drug. Fluorescent rhodamine-labelled nano-micelles distribute with a volume of infusion/volume of distribution (Vi/Vd) ratio of four and five respectively after administration by CED. Administration was not associated with any toxicity when compared to controls. CED of panobinostat-loaded nano-micelles was associated with significantly improved survival when compared to controls (n=8 per group; log-rank test,
<0.001). One hundred percent of treated animals survived the 60-day experimental period and had tumour response on post-mortem histological examination.
CED of nano-micellar panobinostat represents a potential novel therapeutic option for malignant glioma and warrants translation into the clinic.
The ability of sensory networks to transiently store information on the scale of seconds can confer many advantages in processing time-varying stimuli. How a network could store information on such ...intermediate time scales, between typical neurophysiological time scales and those of long-term memory, is typically attributed to persistent neural activity. An alternative mechanism which might allow for such information storage is through temporary modifications to the neural connectivity which decay on the same second-long time scale as the underlying memories. Earlier work that has explored this method has done so by emphasizing one attractor from a limited, pre-defined set. Here, we describe an alternative, a Transient Attractor network, which can learn any pattern presented to it, store several simultaneously, and robustly recall them on demand using targeted probes in a manner reminiscent of Hopfield networks. We hypothesize that such functionality could be usefully embedded within sensory cortex, and allow for a flexibly-gated short-term memory, as well as conferring the ability of the network to perform automatic de-noising, and separation of input signals into distinct perceptual objects. We demonstrate that the stored information can be refreshed to extend storage time, is not sensitive to noise in the system, and can be turned on or off by simple neuromodulation. The diverse capabilities of transient attractors, as well as their resemblance to many features observed in sensory cortex, suggest the possibility that their actions might underlie neural processing in many sensory areas.
A variety of extraimmune system factors, including hormones, play a critical role in regulating immunity. Progesterone has been shown to affect immunity in rodents and humans, mainly at ...concentrations commensurate with pregnancy. These effects are primarily mediated via the progesterone receptor (PR), which acts as a transcription factor, although non-genomic effects of PR activation have been reported. In this study, we evaluated the effects of progesterone on rat dendritic cells (DCs) at ranges encompassing physiologic and pharmacologic concentrations to determine whether progesterone plays a role in modulating DC-mediated immune responses. DCs were derived by culturing rat bone marrow cells in granulocyte macrophage colony-stimulating factor and IL-4. Cells were analyzed for expression of PR using FACS analysis, real-time reverse transcriptase–PCR and fluorescent microscopy. Progesterone treatment of LPS-activated, mature bone marrow-derived dendritic cells (BMDCs) suppressed production of the pro-inflammatory response-promoting cytokines tumor necrosis factor-α and IL-1β in a dose-dependent manner but did not affect production of the pro-inflammatory response-inhibiting cytokine IL-10. Treatment of cells with progesterone also resulted in down-regulation of co-stimulatory molecule CD80 and MHC class II molecule RT1B expression. In addition, progesterone inhibited DC-stimulated proliferation of T cells. Suppression of pro-inflammatory response-promoting cytokine production by progesterone was prevented using the PR antagonist RU486. There was no dose-dependent effect of progesterone treatment on immature DC capacity to take up antigenic peptide. These data indicate that progesterone directly inhibits mature rat BMDC capacity to drive pro-inflammatory responses. This mechanism could contribute to or account for some of the differential expression of autoimmune/inflammatory disease in females.
Objectives
Water is an essential nutrient for thermoregulation, metabolism, cognition, and overall physiological homeostatic function. However, aging adults display a blunted thirst mechanism and ...subsequently have an increased risk for dehydration or hyponatremia. Fluid consumption behaviors are modifiable and the importance of practicing adequate drinking behaviors for aging adults is amplified during exercise. Identification of aging adult’s hydration beliefs and how they attain hydration advice could provide valuable information into ways to promote better drinking habits to reduce fluid imbalances. Thus, this investigation evaluated the knowledge, beliefs and behaviors of middle-aged cyclists (MA) that were associated with hydration status and drinking behavior, before and during a 164-km mass-participation event (ambient temperature, 33.3±2.8ºC(mean±SD)). Design: This cross-sectional field study retrospectively grouped participants by their second urine specific gravity (U
sg
) measurement of the event morning prior to a mass participation cycling event. U
sg
was assessed via handheld refractometer.
Setting
The Hotter N’ Hell Hundred 164-km cycling event in Wichita Falls, Texas during the month of August.
Participants
36 male recreational cyclists (age, 53±9 y(mean±SD)).
Measurements
Participants were grouped according their urine specific gravity as either slightly hyperhydrated (SH; n=12, U
sg
≤1.014), euhydrated (EUH; n=12, U
sg
, 1.015-1.020), or slightly dehydrated (SD; n=12, U
sg
≥1.021). Exercise histories and questionnaires were recorded 24-48 h prior to the cycling event.
Results
Regardless of pre-event hydration status, all groups experienced a similar body mass loss during the 164-km event and finished with statistically similar exercise times; also, drinking behavior within all groups was influenced by multiple factors. The primary factors associated with MA cyclist drinking behavior were trial and error/personal history and thirst; further, the majority of cyclists (≥65%) in SH, EUH, and SD believed that dehydration affects performance negatively. The least important factors included rehydration recommendations from scientific and sports medicine organizations, plus information from sports drink manufacturers.
Conclusion
Considering the complexity of the present findings and the physiological changes that accompany aging such as delayed thirst perception, we recommend that MA cyclists formulate an individualized drinking plan that is based on observations during exercise.
Everyone can recite methodological “urban legends” that were taught in graduate school, learned over the years through experience publishing, or perhaps just heard through the grapevine. In this ...article, the authors trace four widely cited and reported cutoff criteria to their (alleged) original sources to determine whether they really said what they are cited as having said about the cutoff criteria, and if not, what the original sources really said. The authors uncover partial truths in tracing the history of each cutoff criterion and in the end endorse a set of 12 specific guidelines for effective academic referencing provided by Harzing that, if adopted, should help prevent the further perpetuation of methodological urban legends.
Animal and human gene therapy studies utilizing AAV vectors have shown that immune responses to AAV capsid proteins can severely limit transgene expression. The main source of capsid antigen is that ...associated with the AAV vectors, which can be reduced by stringent vector purification. A second source of AAV capsid proteins is that expressed from cap genes aberrantly packaged into AAV virions during vector production. This antigen source can be eliminated by the use of a cap gene that is too large to be incorporated into an AAV capsid, such as a cap gene containing a large intron (captron gene). Here, we investigated the effects of elimination of cap gene transfer and of vector purification by CsCl gradient centrifugation on AAV vector immunogenicity and expression following intramuscular injection in dogs. We found that both approaches reduced vector immunogenicity and that combining the two produced the lowest immune responses and highest transgene expression. This combined approach enabled the use of a relatively mild immunosuppressive regimen to promote robust micro-dystrophin gene expression in Duchenne muscular dystrophy-affected dogs. Our study shows the importance of minimizing AAV cap gene impurities and indicates that this improvement in AAV vector production may benefit human applications.
A bidirectional communication exists between the CNS and the immune system. The autonomic nervous system, through neurotransmitters and neuropeptides, works in parallel with the ...hypothalamic-pituitary-adrenal axis through the actions of glucocorticoids to modulate inflammatory events. The immune system, through the action of cytokines and other factors, in turn, activates the CNS to orchestrate negative-feedback mechanisms that keep the immune response in check. Disruption of these interactions has been associated with a number of syndromes including inflammatory, autoimmune, and cardiovascular diseases, metabolic and psychiatric disorders, and the development of shock. The hypothalamic-pituitary-gonadal axis also plays an important part in regulating immunity through the secretion of sex hormones. Although numerous studies have established a role for immunomodulation by estrogen and testosterone, the role of progesterone is less well understood. Progesterone is crucial for reproductive organ development and maintenance of pregnancy, and more recent studies have clearly shown its role as an important immune regulator. The main focus of this review will be about the role of steroid hormones, specifically glucocorticoids and progesterone, in inflammatory responses and infectious diseases and how dysregulation of their actions may contribute to development of autoimmune and inflammatory disease.
Summary
Background
Increased allergenicity of roasted vs. raw peanut has been reported by showing higher IgE binding to roasted peanut extracts.
Objective
To study the effect of roasting on Ara h 1 ...quantification in peanut using a specific monoclonal antibody‐based ELISA, and to compare the Ara h 1 content from different kernel size peanuts from four runner cultivars.
Methods
Raw or oven‐roasted (177°C for 5–30 min) runner peanuts were crushed and extracted at 60°C. Inhibition ELISA was used to study binding of Ara h 1 purified from raw or roasted peanut. Runner peanuts of four different cultivars were collected, shelled, sized and roasted for 15 min at 177°C. Ara h 1 in the extracts was compared by ELISA.
Results
Ara h 1 levels were up to 22‐fold higher in roasted than in raw peanuts (820 vs. 37 μg/mL, in a representative experiment) with an Ara h 1 peak at 10–15 min of roasting. Inhibition ELISA indicated that this increase was not due to conformational changes in the Ara h 1 monoclonal antibody epitopes. Ara h 1 was found at lower levels in number 1 than in jumbo‐ and medium‐sized peanuts, and no differences were found among cultivars.
Conclusion
These results suggest that roasting increases the efficiency of Ara h 1 extraction, and/or that the monoclonal antibody binding epitopes were more accessible in roasted peanut. Expression of Ara h 1 is associated with peanut maturity.
Steroid hormones, such as progesterone, are able to modify immunity and influence disease outcome. Dendritic cells (DCs) drive potent immune responses, express receptors for steroid hormones, and may ...be a primary target of steroid hormone actions during infection of the genital tract, including uterine tissue. Here, we report that progesterone limited DC-associated activation marker expression and inhibited cytokine secretion by uterine DCs, which was associated with changes in signal transducer and activator of transcription 1 (STAT1) activity. We also found that DCs from mice at stages with higher progesterone concentrations (diestrus, metaestrus) were more sensitive to progesterone than those in stages with lower progesterone concentrations (proestrus, estrus), both in vitro and in vivo. This difference correlated with the levels of progesterone receptor expressed by DCs. These data suggest that progesterone regulates DC function and could contribute to the susceptibility of females to uterine and other genital tract infections at selected time periods throughout the life cycle.
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