Adipocyte-Specific Laminin Alpha 4 Deletion Preserves Adipose Tissue Health despite Increasing Adiposity.

Laminins are heterotrimeric glycoproteins with structural and functional roles in basement membranes. The predominant laminin alpha chain found in adipocyte basement membranes is laminin α4 (LAMA4). Global LAMA4 deletion in mice leads to reduced adiposity and increased energy expenditure, but also results in vascular defects that complicate the interpretation of metabolic data. Here, we describe the generation and initial phenotypic analysis of an adipocyte-specific LAMA4 knockout mouse (Lama4AKO). We first performed an in-silico analysis to determine the degree to which laminin α4 was expressed in human and murine adipocytes. Next, male Lama4AKO and control mice were fed chow or high-fat diets and glucose tolerance was assessed along with serum insulin and leptin levels. Adipocyte area was measured in both epididymal and inguinal white adipose tissue (eWAT and iWAT, respectively), and eWAT was used for RNA-sequencing. We found that laminin α4 was highly expressed in human and murine adipocytes. Further, chow-fed Lama4AKO mice are like control mice in terms of body weight, body composition, and glucose tolerance, although they have larger eWAT adipocytes and lower insulin levels. High-fat-fed Lama4AKO mice are fatter and more glucose tolerant when compared to control mice. Transcriptionally, the eWAT of high-fat fed Lama4AKO mice resembles that of chow-fed control mice. We conclude from these findings that adipocyte-specific LAMA4 deletion is protective in an obesogenic environment, even though overall adiposity is increased.

Oncostatin M Induces Lipolysis and Suppresses Insulin Response in 3T3-L1 Adipocytes.

Oncostatin M (OSM) is an immune cell-derived cytokine that is upregulated in adipose tissue in obesity. Upon binding its receptor (OSMR), OSM induces the phosphorylation of the p66 subunit of Src homology 2 domain-containing transforming protein 1 (SHC1), called p66Shc, and activates the extracellular signal-related kinase (ERK) pathway. Mice with adipocyte-specific OSMR deletion (OsmrFKO) are insulin resistant and exhibit adipose tissue inflammation, suggesting that intact adipocyte OSM-OSMR signaling is necessary for maintaining adipose tissue health. How OSM affects specific adipocyte functions is still unclear. Here, we examined the effects of OSM on adipocyte lipolysis. We treated 3T3-L1 adipocytes with OSM, insulin, and/or inhibitors of SHC1 and ERK and measured glycerol release. We also measured phosphorylation of p66Shc, ERK, and insulin receptor substrate-1 (IRS1) and the expression of lipolysis-associated genes in OSM-exposed 3T3-L1 adipocytes and primary adipocytes from control and OsmrFKO mice. We found that OSM induces adipocyte lipolysis via a p66Shc-ERK pathway and inhibits the suppression of lipolysis by insulin. Further, OSM induces phosphorylation of inhibitory IRS1 residues. We conclude that OSM is a stimulator of lipolysis and inhibits adipocyte insulin response. Future studies will determine how these roles of OSM affect adipose tissue function in health and disease.

Coercion and the Inpatient Treatment Alliance.

OBJECTIVE: The importance of building a strong treatment alliance is widely accepted and uncontroversial. Quantitative research suggests that coercive experiences during psychiatric treatment negatively affect the treatment alliance, but reveals little about how this happens or how patients navigate treatment relationships while experiencing coercion during psychiatric treatment. METHODS: Fifty psychiatric inpatients were interviewed at two hospitals. Patients were asked open-ended questions about the relationship between the treatment alliance and a set of coercive treatment experiences (court-mandated treatment, involuntary hospitalization, locked facilities) and whether such hospital experiences affected the patients' plans for future adherence. Interviews were audio-recorded, transcribed, and qualitatively analyzed. RESULTS: Many participants reported events where coercion made it difficult to form a treatment alliance. An imbalance of power, lack of control, and insufficient participation in treatment planning were described as experiences that interfered with the treatment alliance. Other participants felt the treatment alliance was maintained despite coercive experiences and spoke of good communication with the psychiatrist, understanding the rationale behind interventions, and feeling the psychiatrist was trying to keep the patient's best interests in mind. CONCLUSIONS: Coercive experiences remain undesirable and are frequently detrimental to the treatment alliance. Nevertheless, patients and clinicians should continue to seek a strong treatment alliance even when treatment plans include coercive elements. Efforts to improve communication, to explain the rationale for treatment plans, and to show that clinicians are trying to act in the patient's best interests may help to preserve a therapeutic alliance.

Oncostatin M Mediates Adipocyte Expression and Secretion of Stromal-Derived Factor 1.

Adipose tissue homeostasis depends on interactions between stromal cells, adipocytes, and the cytokines and chemokines they produce. The gp130 cytokine, oncostatin M (OSM), plays a role in adipose tissue homeostasis. Mice, lacking the OSM receptor (OSMR) in adipocytes (OsmrFKO mice), exhibit derangements in adipose tissue, insulin sensitivity, and immune cell balance. Here, we describe a possible role for the chemokine stromal-derived factor 1 (SDF-1) in these alterations. We treated 3T3-L1 adipocytes with OSM and observed a suppression of SDF-1 gene expression and protein secretion, an effect which was partially blunted by OSMR knockdown. However, OsmrFKO mice also exhibited decreased SDF-1 gene and protein expression in adipose tissue. These contrasting results suggest that the loss of adipocyte OSM⁻OSMR signaling in vivo may be indirectly affecting adipokine production and secretion by altering OSM target genes to ultimately decrease SDF-1 expression in the OsmrFKO mouse. We conclude that adipocyte OSM⁻OSMR signaling plays a role in adipose tissue SDF-1 production and may mitigate its effects on adipose tissue homeostasis.

Adipose Tissue Dysfunction Occurs Independently of Obesity in Adipocyte-Specific Oncostatin Receptor Knockout Mice.

OBJECTIVE: This study examined the phenotypic effects of adipocyte-specific oncostatin M receptor (OSMR) loss in chow-fed mice. METHODS: Chow-fed adipocyte-specific OSMR knockout (FKO) mice and littermate OSMRfl/fl controls were studied. Tissue weights, insulin sensitivity, adipokine production, and stromal cell immunophenotypes were assessed in epididymal fat (eWAT); serum adipokine production was also assessed. In vitro, adipocytes were treated with oncostatin M, and adipokine gene expression was assessed. RESULTS: Body weights, fasting blood glucose levels, and eWAT weights did not differ between genotypes. However, the eWAT of OSMRFKO mice was modestly less responsive to insulin stimulation than that of OSMRfl/fl mice. Notably, significant increases in adipokines, including C-reactive protein, lipocalin 2, intercellular adhesion molecule-1, and insulinlike growth factor binding protein 6, were observed in the eWAT of OSMRFKO mice. In addition, significant increases in fetuin A and intercellular adhesion molecule-1 were detected in OSMRFKO serum. Flow cytometry revealed a significant increase in leukocyte number and modest, but not statistically significant, increases in B cells and T cells in the eWAT of OSMRFKO mice. CONCLUSIONS: The chow-fed OSMRFKO mice exhibited adipose tissue dysfunction and increased proinflammatory adipokine production. These results suggest that intact adipocyte oncostatin M-OSMR signaling is necessary for adipose tissue immune cell homeostasis.

Adipogenic Differentiation of Human Adipose-Derived Stem Cells Grown as Spheroids.

Understanding the process of adipogenesis is critical if suitable therapeutics for obesity and related metabolic diseases are to be found. The current study presents proof of feasibility of creating a 3-D spheroid model using human adipose-derived stem cells (hASCs) and their subsequent adipogenic differentiation. hASC spheroids were formed atop an elastin-like polypeptide-polyethyleneimine (ELP-PEI) surface and differentiated using an adipogenic cocktail. Spheroids were matured in the presence of dietary fatty acids (linoleic or oleic acid) and evaluated based on functional markers including intracellular protein, CD36 expression, triglyceride accumulation, and PPAR-γ gene expression. Spheroid size was found to increase as the hASCs matured in the adipocyte maintenance medium, though the fatty acid treatment generally resulted in smaller spheroids compared to control. A stable protein content over the 10-day maturation period indicated contact-inhibited proliferation as well as minimal loss of spheroids during culture. Spheroids treated with fatty acids showed greater amounts of intracellular triglyceride content and greater expression of the key adipogenic gene, PPAR-γ. We also demonstrated that 3-D spheroids outperformed 2-D monolayer cultures in adipogenesis. We then compared the adipogenesis of hASC spheroids to that in 3T3-L1 spheroids and found that the triglyceride accumulation was less profound in hASC spheroids than that in 3T3-L1 adipocytes, correlated with smaller average spheroids, suggesting a relatively slower differentiation process. Taken together, we have shown the feasibility of adipogenic differentiation of patient-derived hASC spheroids, which with further development, may help elucidate key features in the adipogenesis process.

A Review of HIV Pre-Exposure Prophylaxis: The Female Perspective.

When taken consistently, pre-exposure prophylaxis (PrEP) against human immunodeficiency virus (HIV) with once daily tenofovir disoproxil fumarate-emtricitabine (TDF-FTC) has been shown to safely reduce the incidence of HIV infection in high-risk individuals by more than 90%. Yet, according to the Centers for Disease Control and Prevention, there were about 2.1 million new cases of HIV reported worldwide in 2015. Undoubtedly, there is significant room for improvement to prevent the transmission of HIV. Research to date has been heavily focused on the high-risk men who have sex with men (MSM) population, yet, many women worldwide remain at high risk of HIV transmission. PrEP offers women a protection method that is discrete, does not require partner consent, and may be compatible with both contraception or conception as desired. However, women often remain under-represented in HIV prevention literature and are reported to have lower real-world uptake in comparison to men. Furthermore, clinical trials that do focus on the female population demonstrate mixed efficacy results that highlight the adherence challenges in this population. It is essential to identify factors that contribute to PrEP non-adherence as well as barriers to preventative treatment. This review will discuss the clinical evidence behind PrEP in women, current barriers to use afflicting this population, pharmacotherapy considerations for the female patient, alternative and future agents, and the current real-world application of PrEP.

HIV and STI knowledge, testing, and risk among adult crack users in Mexico city: baseline findings from a feasibility study.

Recent research has documented crack cocaine's increasing spread in Mexico, which is likely to contribute to the rapid transmission of HIV and other sexually transmitted infections (STIs). In Mexico, crack use is increasing most rapidly in vulnerable, hard-to-reach populations, where little is known about risk behaviors. This report aims to present baseline data regarding HIV and STI knowledge and testing prevalence from an innovative projection mapping HIV intervention, in which 3-D illusions, animation, and visual text graphics and sound are projected onto buildings with health messages that were designed to disrupt everyday life and connect with the target population. Fifty-eight men and women who used crack in the past month without receiving drug treatment were recruited and interviewed before the projection mapping intervention took place. Testing instruments included a sociodemographic assessment, drug use and treatment profile, HIV and STI knowledge questionnaires, and a sex and drug risk assessment. The mean scores for respondents on the HIV Knowledge Questionnaire (10.5 out of 18, 58.3%) and STD Knowledge Questionnaire (9.5 out of 27, 35.2%) were both low. Respondents also reported high rates of sexual risk behaviors, with 73% reporting never using a condom and 64% never being tested for HIV. This report provides a portrait of STI and HIV risk among a vulnerable population in Mexico City and the need for urgent interventions to prevent the spread of STIs and HIV. The associated projection mapping intervention will seek to increase HIV and STI knowledge and reduce risk in this hard-to-reach population.

Loss of Oncostatin M Signaling in Adipocytes Induces Insulin Resistance and Adipose Tissue Inflammation in Vivo.

Oncostatin M (OSM) is a multifunctional gp130 cytokine. Although OSM is produced in adipose tissue, it is not produced by adipocytes. OSM expression is significantly induced in adipose tissue from obese mice and humans. The OSM-specific receptor, OSM receptor ß (OSMR), is expressed in adipocytes, but its function remains largely unknown. To better understand the effects of OSM in adipose tissue, we knocked down Osmr expression in adipocytes in vitro using siRNA. In vivo, we generated a mouse line lacking Osmr in adiponectin-expressing cells (OSMR(FKO) mice). The effects of OSM on gene expression were also assessed in vitro and in vivo OSM exerts proinflammatory effects on cultured adipocytes that are partially rescued by Osmr knockdown. Osm expression is significantly increased in adipose tissue T cells of high fat-fed mice. In addition, adipocyte Osmr expression is increased following high fat feeding. OSMR(FKO) mice exhibit increased insulin resistance and adipose tissue inflammation and have increased lean mass, femoral length, and bone volume. Also, OSMR(FKO) mice exhibit increased expression of Osm, the T cell markers Cd4 and Cd8, and the macrophage markers F4/80 and Cd11c Interestingly, the same proinflammatory genes induced by OSM in adipocytes are induced in the adipose tissue of the OSMR(FKO) mouse, suggesting that increased expression of proinflammatory genes in adipose tissue arises both from adipocytes and other cell types. These findings suggest that adipocyte OSMR signaling is involved in the regulation of adipose tissue homeostasis and that, in obesity, OSMR ablation may exacerbate insulin resistance by promoting adipose tissue inflammation.

Probiotics for disease prevention: a focus on ventilator-associated pneumonia.

OBJECTIVE: To summarize published data regarding the safety and efficacy of probiotics in the prevention of ventilator-associated pneumonia (VAP). DATA SOURCES: PubMed databases (January 2000-August 2011) were searched and a bibliographic review of published articles was performed to identify original reports of probiotic administration for the prevention of VAP using the search terms probiotics, synbiotics, and ventilator-associated pneumonia. STUDY SELECTION AND DATA EXTRACTION: Two pilot studies, 2 randomized controlled trials (RCTs), and 1 meta-analysis have addressed probiotic use for VAP prevention and were included in the review. DATA SYNTHESIS: VAP frequently occurs in mechanically ventilated patients. Given the lack of new antimicrobial agents, probiotics have been studied for their ability to modify human microflora colonization. Two studies examining pathogen colonization rates favored probiotics, with reduced incidence and increased duration until the emergence of new species. One prospective RCT found significant reduction in the incidence of VAP and colonization rates, but no significant difference in patient disposition outcomes. Another RCT examining 28-day mortality found no overall benefit with probiotic use and no reduction in colonization rates. CONCLUSIONS: Clinical trials have failed to demonstrate a consistent beneficial effect of probiotics in mechanically ventilated patients; thus, they are not recommended for routine clinical use. However, heterogeneity among study designs may hinder this assessment and the designs should be unified in future research.

Primary abdominal gestational choriocarcinoma.

BACKGROUND: Gestational choriocarcinoma associated with ectopic pregnancy is an extremely infrequent event. Primary abdominal gestational choriocarcinoma has not been previously described. CASE: A pregnant woman presented to the emergency room with 6 days of vaginal spotting. Her last menstrual period suggested a gestation at 6 4/7 weeks. Transvaginal sonogram showed a hemoperitoneum with no intrauterine pregnancy. The serum human chorionic gonadotropin level was noted to be 317,735 mIU/mL. A 20 x 20-mm friable, bleeding mass on the left anterior abdominal wall was laparoscopically resected. Gestational choriocarcinoma was identified on histopathologic review. International Federation of Gynecology and Obstetrics stage IV:4 was assigned, and the patient achieved clinical remission with combination chemotherapy. CONCLUSION: Primary abdominal gestational choriocarcinoma can present with findings similar to a ruptured ectopic pregnancy; it should be treated by surgical excision and chemotherapy.

Inflammation, immune reactivity, and angiogenesis in a severe combined immunodeficiency model of rheumatoid arthritis.

Severe combined immunodeficiency (SCID) mice were engrafted with rheumatoid arthritis (RA) synovium and evaluated to determine whether RA synovial morphology and function were maintained in the RA-SCID grafts. The four major components of RA synovitis, inflammation, immune reactivity, angiogenesis, and synovial hyperplasia persisted in RA-SCID grafts for 12 weeks. Retention of chronic inflammatory infiltrates was demonstrated by histological evaluation and by immunohistology for CD3, CD20, and CD68. Staining for CD68 also revealed that the grafts had undergone reorganization of the tissue, possibly as a result of fibroblast hyperplasia. Immune and inflammatory components were confirmed by the detection of human immunoglobulins and human interleukin-6 in serum samples obtained from grafted animals. Human blood vessels were detected by dense expression of CD31. Small vessels persistently expressed the vitronectin receptor, alpha v beta 3, a marker of angiogenesis. All vessels expressed VAP-1, a marker of activated endothelial cells. Finally, the grafts retained the ability to support immigration by human leukocytes, as demonstrated by the functional capacity to recruit adoptively transferred 5- (and -6)-carboxyfluorescein diacetate succinimidyl ester-labeled T cells. T cells entering the RA-SCID grafts became activated and produced interferon-gamma, as detected by reverse transcriptase-polymerase chain reaction analysis. These studies demonstrate that the RA-SCID model maintains many of the phenotypic and functional features of the inflamed RA synovium.