Sustained release of drug-loaded nanoparticles from injectable hydrogels enables long-term control of macrophage phenotype.

Injectable hydrogels may be pre-formed through dynamic crosslinks, allowing for injection and subsequent retention in the tissue by shear-thinning and self-healing processes, respectively. These properties enable the site-specific delivery of encapsulated therapeutics; yet, the sustained release of small-molecule drugs and their cell-targeted delivery remains challenging due to their rapid diffusive release and non-specific cellular biodistribution. Herein, we develop an injectable hydrogel system composed of a macrophage-targeted nanoparticle (cyclodextrin nanoparticles, CDNPs) crosslinked by adamantane-modified hyaluronic acid (Ad-HA). The polymer-nanoparticle hydrogel uniquely leverages cyclodextrin's interaction with small molecule drugs to create a spatially discrete drug reservoir and with adamantane to yield dynamic, injectable hydrogels. Through an innovative two-step drug screening approach and examination of 45 immunomodulatory drugs with subsequent in-depth transcriptional profiling of both murine and human macrophages, we identify celastrol as a potent inhibitor of pro-inflammatory (M1-like) behavior that furthermore promotes a reparatory (M2-like) phenotype. Celastrol encapsulation within the polymer-nanoparticle hydrogels permitted shear-thinning injection and sustained release of drug-laden nanoparticles that targeted macrophages to modulate cell behavior for greater than two weeks in vitro. The modular hydrogel system is a promising approach to locally modulate cell-specific phenotype in a range of applications for immunoregenerative medicine.

Pro-inflammatory polarization primes Macrophages to transition into a distinct M2-like phenotype in response to IL-4.

Tissue repair is largely regulated by diverse Mϕ populations whose functions are timing- and context-dependent. The early phase of healing is dominated by pro-inflammatory Mϕs, also known as M1, followed by the emergence of a distinct and diverse population that is collectively referred to as M2. The extent of the diversity of the M2 population is unknown. M2 Mϕs may originate directly from circulating monocytes or from phenotypic switching of pre-existing M1 Mϕs within the site of injury. The differences between these groups are poorly understood, but have major implications for understanding and treating pathologies characterized by deficient M2 activation, such as chronic wounds, which also exhibit diminished M1 Mϕ behavior. This study investigated the influence of prior M1 activation on human Mϕ polarization to an M2 phenotype in response to IL-4 treatment in vitro. Compared to unactivated (M0) Mϕs, M1 Mϕs up-regulated several receptors that promote the M2 phenotype, including the primary receptor for IL-4. M1 Mϕs also up-regulated M2 markers in response to lower doses of IL-4, including doses as low as 10 pg/mL, and accelerated STAT6 phosphorylation. However, M1 activation appeared to also change the Mϕ response to treatment with IL-4, generating an M2-like phenotype with a distinct gene and protein expression signature compared to M2 Mϕs prepared directly from M0 Mϕs. Functionally, compared to M0-derived M2 Mϕs, M1-derived M2 Mϕs demonstrated increased migratory response to SDF-1α, and conditioned media from these Mϕs promoted increased migration of endothelial cells in transwell assays, although other common Mϕ-associated functions such as phagocytosis were not affected by prior polarization state. In summary, M1 polarization appears to prime Mϕs to transition into a distinct M2 phenotype in response to IL4, which leads to increased expression of some genes and proteins and decreased expression of others, as well as functional differences. Together, these findings indicate the importance of prior M1 activation in regulating subsequent M2 behavior, and suggest that correcting M1 behavior may be a therapeutic target in dysfunctional M2 activation.

Draft Genome Sequences of Bacillus glennii V44-8, Bacillus saganii V47-23a, Bacillus sp. Strain V59.32b, Bacillus sp. Strain MER_TA_151, and Paenibacillus sp. Strain MER_111, Isolated from Cleanrooms Where the Viking and Mars Exploration Rover Spacecraft Were Assembled.

We report the draft genome sequences of Bacillus glennii V44-8, Bacillus saganii V47-23a, and Bacillus sp. strain V59.32b, isolated from the Viking spacecraft assembly cleanroom, and Bacillus sp. strain MER_TA_151 and Paenibacillus sp. strain MER_111, isolated from the Mars Exploration Rover (MER) assembly cleanroom.

Anti-CTLA-4 Activates Intratumoral NK Cells and Combined with IL15/IL15Rα Complexes Enhances Tumor Control.

Antibodies targeting CTLA-4 induce durable responses in some patients with melanoma and are being tested in a variety of human cancers. However, these therapies are ineffective for a majority of patients across tumor types. Further understanding the immune alterations induced by these therapies may enable the development of novel strategies to enhance tumor control and biomarkers to identify patients most likely to respond. In several murine models, including colon26, MC38, CT26, and B16 tumors cotreated with GVAX, anti-CTLA-4 efficacy depends on interactions between the Fc region of CTLA-4 antibodies and Fc receptors (FcR). Anti-CTLA-4 binding to FcRs has been linked to depletion of intratumoral T regulatory cells (Treg). In agreement with previous studies, we found that Tregs infiltrating CT26, B16-F1, and autochthonous Braf V600E Pten -/- melanoma tumors had higher expression of surface CTLA-4 (sCTLA-4) than other T-cell subsets, and anti-CTLA-4 treatment led to FcR-dependent depletion of Tregs infiltrating CT26 tumors. This Treg depletion coincided with activation and degranulation of intratumoral natural killer cells. Similarly, in non-small cell lung cancer (NSCLC) and melanoma patient-derived tumor tissue, Tregs had higher sCTLA-4 expression than other intratumoral T-cell subsets, and Tregs infiltrating NSCLC expressed more sCTLA-4 than circulating Tregs. Patients with cutaneous melanoma who benefited from ipilimumab, a mAb targeting CTLA-4, had higher intratumoral CD56 expression, compared with patients who received little to no benefit from this therapy. Furthermore, using the murine CT26 model we found that combination therapy with anti-CTLA-4 plus IL15/IL15Rα complexes enhanced tumor control compared with either monotherapy.

Sequential drug delivery to modulate macrophage behavior and enhance implant integration.

Macrophages are major upstream regulators of the inflammatory response to implanted biomaterials. Sequential functions of distinct macrophage phenotypes are essential to the normal tissue repair process, which ideally results in vascularization and integration of implants. Improper timing of M1 or M2 macrophage activation results in dysfunctional healing in the form of chronic inflammation or fibrous encapsulation of the implant. Thus, biphasic drug delivery systems that modulate macrophage behavior are an appealing approach to promoting implant integration. In this review, we describe the timing and roles of macrophage phenotypes in healing, then highlight current drug delivery systems designed to sequentially modulate macrophage behavior.

Insomnia is associated with impaired quality of life among bariatric surgery candidates.

SUMMARY: Little is known about the prevalence or impact of insomnia symptoms in obese individuals pursuing bariatric surgery. The present study from the Rhode Island Bariatric Surgery (RIBS) project examined insomnia symptoms among 2300 individuals pursuing bariatric surgery. Patients were evaluated using the Structured Clinical Interview for DSM-IV Disorders (SCID), Schedule for Affective Disorders (SADS), Rhode Island Bariatric Surgery Interview (RIBSI), and the SF-36 as a measure of quality of life. The presence of insomnia symptoms was determined via ratings for the SCID items assessing initial, middle, and terminal insomnia symptoms, and the SADS insomnia item was used to measure severity of insomnia symptoms. Clinical and demographic variables were obtained from the SCID and self-report measures. Insomnia symptoms were endorsed by 25.8% of participants. Bariatric patients with insomnia symptoms were rated as having a more severe clinical presentation and lower functioning, and were more likely to have a history of psychiatric treatment and/or hospitalization, compared to bariatric patients without insomnia. Linear regression analyses demonstrated that insomnia severity was a significant predictor for scores on each of the 8 SF-36 subscales after accounting for age, gender, race, education level, BMI, depression severity, and sleep apnea. Additionally, a multivariate analysis of covariance (MANCOVA) controlling for depression severity and sleep apnea demonstrated significantly poorer scores on 6 of the 8 SF-36 subscales for bariatric patients with current insomnia symptoms. Results revealed that insomnia symptoms are common among bariatric patients and are associated with reduced quality of life and poorer current functioning. This suggests that insomnia symptoms are an important clinical target in bariatric patients prior to surgery.:

Perylenequinones: Isolation, Synthesis, and Biological Activity.

The perylenequinones are a novel class of natural products characterized by pentacyclic conjugated chromophore giving rise to photoactivity. Potentially useful light-activated biological activity, targeting protein kinase C (PKC), has been identified for several of the natural products. Recently discovered new members of this class of compound, as well as several related phenanthroperylenequinones, are reviewed. Natural product modifications that improve biological profiles, and avenues for the total synthesis of analogs, which are not available from the natural product series, are outlined. An overview of structure/function relationships is provided.

Intraindividual variability in daily sleep and pain ratings among chronic pain patients: bidirectional association and the role of negative mood.

OBJECTIVES: Sleep disturbance is a common problem among chronic pain patients. Cross-sectional data from clinical populations and experimental studies have shown an association between sleep disturbance and pain. However, there has been little prospective research into the relationship between daily variability between sleep and pain among chronic pain patients. METHODS: Twenty-two women with chronic pain (back pain, facial pain, fibromyalgia) completed a sleep diary and wore an actigraph for a 2-week period. Self-report measures of pain, mood, and sleep were also completed at baseline. Hierarchical linear modeling (HLM) was used to examine intraindividual variability in sleep and pain ratings among these women. The impact of mood and baseline pain ratings was also examined as potential moderators. RESULTS: Hierarchical linear modeling analyses supported a bidirectional relationship between sleep and pain, such that a night of poor sleep was followed by increased pain ratings the following day and a day of increased pain was followed by a night of poor sleep. Depression scores further influenced these relationships. DISCUSSION: Prospective examination supported a bidirectional relationship between sleep and pain among a group of women with chronic pain. Depressive symptoms had a moderating impact on these relationships. These findings suggest that addressing sleep is important in the treatment of individuals with chronic pain.

Severe insomnia is associated with more severe presentation and greater functional deficits in depression.

Depression is among the most common reasons for seeking psychiatric treatment, and insomnia symptoms are common in the clinical picture of depression. The present study examines the clinical presentation and psychosocial functioning among depressed outpatients with severe symptoms of insomnia in comparison to depressed outpatients without severe insomnia symptoms. The present sample included 2900 treatment-seeking individuals, with 1057 patients having a principal diagnosis of major depressive disorder (MDD). All patients were evaluated using the Structured Clinical Interview for DSM-IV Disorders (SCID), Schedule for Affective Disorders (SADS), and self-report measures of mood and psychosocial functioning. SADS Insomnia ratings were used to determine the presence of severe insomnia symptoms. Clinical, demographic, and psychosocial variables were obtained from the SCID and self-report measures. Among the patients with MDD, 24.7% endorsed severe insomnia symptoms. These individuals were older at time of presentation, were less likely to be married, had a lower education level, had a longer duration of the current depressive episode, were rated as more severe on the CGI, had poorer current functioning via GAF, and had higher HAM-D 21 scores. After controlling for severity, MDD patients with severe insomnia symptoms had poorer social functioning over the past 5 years, though this did not reach the significance level of p < .01, and significantly lower scores on 3 of the 8 SF-36 subscales (p < 0.01). These findings indicate that severe insomnia symptoms are associated with poorer psychosocial functioning and a more severe clinical presentation in patients with MDD. This argues for addressing severe insomnia symptoms among depressed patients, either via behavioral treatment or pharmacologic treatment options.

Characterization of the apoptotic response of human leukemia cells to organosulfur compounds.

BACKGROUND: Novel therapeutic agents that selectively induce tumor cell death are urgently needed in the clinical management of cancers. Such agents would constitute effective adjuvant approaches to traditional chemotherapy regimens. Organosulfur compounds (OSCs), such as diallyl disulfide, have demonstrated anti-proliferative effects on cancer cells. We have previously shown that synthesized relatives of dysoxysulfone, a natural OSC derived from the Fijian medicinal plant, Dysoxylum richi, possess tumor-specific antiproliferative effects and are thus promising lead candidates. METHODS: Because our structure-activity analyses showed that regions flanking the disulfide bond mediated specificity, we synthesized 18 novel OSCs by structural modification of the most promising dysoxysulfone derivatives. These compounds were tested for anti-proliferative and apoptotic activity in both normal and leukemic cells. RESULTS: Six OSCs exhibited tumor-specific killing, having no effect on normal bone marrow, and are thus candidates for future toxicity studies. We then employed mRNA expression profiling to characterize the mechanisms by which different OSCs induce apoptosis. Using Gene Ontology analysis we show that each OSC altered a unique set of pathways, and that these differences could be partially rationalized from a transcription factor binding site analysis. For example, five compounds altered genes with a large enrichment of p53 binding sites in their promoter regions (p < 0.0001). CONCLUSIONS: Taken together, these data establish OSCs derivatized from dysoxysulfone as a novel group of compounds for development as anti-cancer agents.

Negative mood mediates the effect of poor sleep on pain among chronic pain patients.

OBJECTIVES: Sleep disturbances and increased negative mood are common among chronic pain patients. Research suggests that sleep disruption can contribute to increased pain; however, the role of negative mood in this relationship is unclear. The present study investigated the relationship among sleep disturbance, negative mood, and pain within a large sample of chronic pain patients. It was hypothesized that negative mood would mediate the relationship between sleep and pain. METHODS: In all, 292 chronic pain patients (116 facial pain, 55 back pain, 121 fibromyalgia) between 18 and 65 years of age (M=46.67) were recruited from 3 chronic pain clinics at a large tertiary care hospital. Patients completed validated measures of pain, negative mood, and sleep during a routine clinical assessment. Structural equation modeling examined the relationship between sleep, negative mood, and pain. RESULTS: All 3 groups of patients reported sleep disturbances, with these being highest among back pain and fibromyalgia patients. Structural equation modeling analyses revealed a significant direct relationship between poor sleep and pain, and further demonstrated that negative mood mediated the relationship between poor sleep and pain in this sample of chronic pain patients. DISCUSSION: These findings suggest that addressing negative mood directly, or by addressing sleep disturbances in chronic pain patients, may have a beneficial impact on patients' pain. As sleep disturbance may be causing negative mood, treating the sleep disturbance may also be beneficial among chronic pain patients. Negative mood may perpetuate the impact of sleep disturbances on pain, possibly through increased arousal or disruptions in diurnal patterns.

Synthesis of the cores of hypocrellin and shiraiachrome: diastereoselective 1,8-diketone aldol cyclization.

Intramolecular 1,8-diketone aldol reactions were studied as a tool for the construction of the seven-membered rings of hypocrellin and shiraiachrome. Conditions were identified to obtain the relative stereochemistries present in the two natural products with excellent diastereoselectivity. In addition, a nine-membered ring congener, which has yet to be observed in nature, formed with high selectivity when a hindered amine was used in conjunction with silazide bases.

Perylenequinone natural products: total synthesis of hypocrellin A.

An efficient and stereoselective total synthesis of the perylenequinone natural product hypocrellin A (1) is described. The key features include a potentially biomimetic 1,8-diketone aldol cyclization to set the centrochiral C7,C7'-stereochemistry, bis(trifluoroacetoxy)iodobenzene mediated oxygenation, a palladium-catalyzed decarboxylation, and an enantioselective catalytic oxidative 2-naphthol coupling to establish the biaryl axial chirality. The helical stereochemistry is formed from an axial chiral intermediate and is then utilized in a dynamic stereochemical transfer to dictate the stereochemistry of the C7,C7'-seven membered ring formed during the aldol cyclization.

Perylenequinone natural products: total syntheses of the diastereomers (+)-phleichrome and (+)-calphostin D by assembly of centrochiral and axial chiral fragments.

The first total synthesis of (+)-calphostin D and the total synthesis of (+)-phleichrome are outlined. The convergent syntheses utilize an enantiopure biaryl common intermediate, which is formed via an enantioselective catalytic biaryl coupling. The established axial chirality is transferred to the perylenequinone helical stereochemistry with good fidelity. Additionally, efforts focused on the installation of the stereogenic C7,C7'-2-hydroxypropyl groups. Three routes were evaluated to establish the C7,C7'-stereochemistry, in which the successful route involved a double epoxide alkylation with a complex axial chiral biscuprate. This strategy not only allowed the synthesis of the unnatural isomers of calphostin D and phleichrome for assessment in biological systems but also provided valuable information for the syntheses of the more complex cercosporin and hypocrellin A.

Patient-centered perspective on treatment outcomes in chronic pain.

OBJECTIVE: To define patient-determined success criteria for fibromyalgia and back pain treatment across four outcome domains: pain, fatigue, emotional distress, interference with daily activities. DESIGN: Retrospective correlational clinical sample design. SETTING: Tertiary care clinics at health science center. PATIENTS: 248 fibromyalgia patients and 52 back pain patients. INTERVENTIONS: N/A. OUTCOME MEASURES: Patient Centered Outcomes Questionnaire, measures of usual pain intensity and pain unpleasantness. RESULTS: Overall, for treatment to be considered successful, fibromyalgia patients required pain levels of 3.30 (54% reduction), fatigue levels of 3.08 (60% reduction), distress levels of 2.49 (60% reduction), and interference levels of 2.67 (63% reduction). Comparatively, back pain patients required pain levels of 2.23 (58% reduction), fatigue levels of 2.29 (57% reduction), distress levels of 1.65 (67% reduction), and interference levels of 1.81 (68% reduction). Overall, both fibromyalgia and back pain patients did not expect to meet their criteria for success. CONCLUSIONS: Results highlight the importance of assessing the patient's view of successful outcome. Both fibromyalgia and back pain patients appear to have stringent criteria for success that existing treatments are often unlikely to meet. Comparison across groups indicated fibromyalgia patients have higher usual levels of pain, fatigue, distress, and interference. Interestingly, fibromyalgia patients also require greater changes across domains in order to consider treatment successful, despite rating higher levels of pain, fatigue, distress, and interference as successful. Recognizing patients' success criteria and treatment expectations encourages discussion and development of individualized treatment goals, and wider implementation of individualized treatment for chronic-pain populations is encouraged.

Tackling sleeplessness: psychological treatment options for insomnia in older adults.

This paper provides a broad review of the extant literature involving the treatment of sleeplessness in older adults with insomnia. First, background information (including information regarding key issues in late-life insomnia and epidemiology of late-life insomnia) pertinent to achieving a general understanding of insomnia in the elderly is presented. Next, theories of insomnia in older adults are examined and discussed in relation to treatment of insomnia in late-life. With a general knowledge base provided, empirical evidence for both pharmacological (briefly) and psychological treatment options for insomnia in late-life are summarized. Recent advances in the psychological treatment of insomnia are provided and future directions are suggested. This review is not meant to be all-inclusive; however, it is meant to provide professionals across multiple disciplines (physicians; psychologists; applied and basic researchers) with a mix of breadth and depth of knowledge related to insomnia in late-life. It is our hope that readers will see the evidence in support of psychological treatments for late-life insomnia, and the utility in continuing to investigate this treatment modality.

Somatic focus/awareness: Relationship to negative affect and pain in chronic pain patients.

Somatic focus refers to the tendency to notice and report physical symptoms, and has been investigated in relation to chronically painful conditions. This study investigated the relationship between somatic focus, as measured by the Pennebaker Inventory of Limbic Languidness (PILL), negative affect and pain. A secondary purpose of the present study was to examine sex differences in these relationships. Participants included 280 chronic pain patients (69.6% females, 88.9% Caucasian), who completed a battery of self-report measures on somatic focus, pain, negative affect, coping, and dysfunction. Results for the overall sample revealed that the PILL shares considerable variance with measures of negative affect, particularly with the physiological components of anxiety and depression. When the results were analyzed separately for male and female patients, it was found that several components of negative affect and cognitive factors play a stronger role in predicting somatic focus among men compared to women. Additional analyses then examined whether somatic focus was predictive of male and female patients' pain reports. Results indicated that somatic focus explained a small, but unique amount of variance in female patients' pain reports, which differed from the relationship observed among male patients.

Development of a catalytic aromatic decarboxylation reaction.

A palladium-catalyzed aromatic decarboxylation reaction has been developed. With electron-rich aromatic acids, the reaction proceeds efficiently under fairly mild conditions in good yields. The method was useful with complex functionalized substrates containing hindered carboxylic acids.

Sleep and risk-taking behavior in adolescents.

The primary purpose of this study was to examine the relationship between adolescents' sleep-wake patterns and risk-taking behavior. A second goal was to replicate the results obtained by Wolfson and Carskadon (1998) regarding adolescents' sleep habits. Three hundred eighty-eight adolescents (217 males, 171 females) completed the Sleep Habits Survey and the Youth Risk Behavior Survey. The results indicated that adolescents who reported longer weekend delay and higher levels of sleep problems also reported significantly higher levels of risk-taking behaviors, and students' weekend delay was also related to their academic performance in this sample. As in the sample studied by Wolfson and Carskadon (1998), the adolescents in this study exhibited changes in both weekday and weekend sleep habits across grade/age. However in the present study, only school-night total sleep time and weekend delay were related to adolescents' daytime functioning, with no significant relationships being found between weekend oversleep and daytime functioning. This provides partial support for the findings of Wolfson and Carskadon (1998). Overall, sleep-wake patterns were found to relate to risk-taking behavior during adolescence in this study.