ABHD6 suppression promotes anti-inflammatory polarization of adipose tissue macrophages via 2-monoacylglycerol/PPAR signaling in obese mice.

OBJECTIVE: Pro-inflammatory polarization of adipose tissue macrophages (ATMs) plays a critical role in the pathogenesis of obesity-associated chronic inflammation. However, little is known about the role of lipids in the regulation of ATMs polarity and inflammation in response to metabolic stress. Deletion of α/ß-hydrolase domain-containing 6 (ABHD6), a monoacylglycerol (MAG) hydrolase, has been shown to protect against diet-induced obesity and insulin resistance. METHODS: Here we investigated the immunometabolic role of macrophage ABHD6 in response to nutrient excess using whole-body ABHD6-KO mice and human and murine macrophage cell-lines treated with KT203, a selective and potent pharmacological ABHD6 inhibitor. RESULTS: KO mice on high-fat diet showed lower susceptibility to systemic diet-induced inflammation. Moreover, in the setting of overnutrition, stromal vascular cells from gonadal fat of KO vs. control mice contained lower number of M1 macrophages and exhibited enhanced levels of metabolically activated macrophages (MMe) and M2 markers, oxygen consumption, and interleukin-6 (IL-6) release. Likewise, under in vitro nutri-stress condition, inhibition of ABHD6 in MMe-polarized macrophages attenuated the expression and release of pro-inflammatory cytokines and M1 markers and induced the upregulation of lipid metabolism genes. ABHD6-inhibited MMe macrophages showed elevated levels of peroxisome proliferator-activated receptors (PPARs) and 2-MAG species. Notably, among different MAG species, only 2-MAG treatment led to increased levels of PPAR target genes in MMe macrophages. CONCLUSIONS: Collectively, our findings identify ABHD6 as a key component of pro-inflammatory macrophage activation in response to excess nutrition and implicate an endogenous macrophage lipolysis/ABHD6/2-MAG/PPARs cascade, as a lipid signaling and immunometabolic pathway, which favors the anti-inflammatory polarization of ATMs in obesity.

Nitrous oxide production from soils amended with biogas residues and cattle slurry.

The amount of residues generated from biogas production has increased dramatically due to the worldwide interest in renewable energy. A common way to handle the residues is to use them as fertilizers in crop production. Application of biogas residues to agricultural soils may be accompanied with environmental risks, such as increased NO emission. In 24-d laboratory experiments, NO dynamics and total production were studied in arable soils (sandy, clay, and organic) amended with one of two types of anaerobically digested biogas residues (BR-A and BR-B) generated from urban and agricultural waste and nondigested cattle slurry (CS) applied at rates corresponding to 70 kg NH-N ha. Total NO-N losses from the sandy soil were higher after amendment with BR-B (0.32 g NO-N m) than BR-A or CS (0.02 and 0.18 g NO-N m, respectively). In the clay soil, NO-N losses were very low for CS (0.02 g NO-N m) but higher for BR-A and BR-B (0.25 and 0.15 g NO-N m, respectively). In the organic soil, CS gave higher total NO-N losses (0.31 g NO-N m) than BR-A or BR-B (0.09 and 0.08 g NO-N m, respectively). Emission peaks differed considerably between soils, occurring on Day 1 in the organic soil and on Days 11 to 15 in the sand, whereas in the clay the peak varied markedly (Days 1, 6, and 13) depending on residue type. In all treatments, NH concentration decreased with time, and NO concentration increased. Potential ammonium oxidation and potential denitrification activity increased significantly in the amended sandy soil but not in the organic soil and only in the clay amended with CS. The results showed that fertilization with BR can increase NO emissions and that the size is dependent on the total N and organic C content of the slurry and on soil type. In conclusion, the two types of BR and the CS are not interchangeable regarding their effects on NO production in different soils, and, hence, matching fertilizer type to soil type could reduce NO emissions. For instance, it could be advisable to avoid fertilization of organic soils with CS containing high amounts or organic C and instead use BR. In clay soil, however, the risk of NO emissions could be lowered by choosing a CS.

Clinicopathological analysis of colorectal cancers with PIK3CA mutations in Middle Eastern population.

Activation of the phosphatidylinositol 3'-kinase (PI3K)/AKT pathway results in an increase in cell proliferation and survival. Somatic mutations within the PI3K catalytic subunit, PIK3CA are common cause of increasing PI3K activity and are believed to be oncogenic in many cancer types. Few reports addressed the association between PIK3CA mutations and tumor progression specifically in microsatellite instable (MSI) colorectal cancer (CRC). In the present study, we have evaluated PIK3CA mutational status in a series of 410 Middle Eastern CRC and 13 colon cell lines to study the prevalence of PIK3CA mutations in MSI cases, PTEN expression in CRC and possibility of therapeutic targeting of this set of patients. PIK3CA mutations were found in four of the cell lines tested and 51 colorectal carcinomas (12.2%). Three of these four mutated cell lines were MSI. PTEN was inactivated in 66.1% of the CRC. Furthermore, we observed a strong association between PIK3CA mutations and MSI status (P=0.0046) while PTEN loss was more frequent in microsatellite stable (MSS) CRC (P=0.043). A high prevalence of genetic alterations in PI3K/AKT pathway in Saudi cohort of CRC, predominance of PIK3CA mutations in the MSI subgroup and their possible involvement in development/progression of this subset of CRC are some of the significant findings of our study.

Genome-wide expression analysis of Middle Eastern papillary thyroid cancer reveals c-MET as a novel target for cancer therapy.

In an attempt to find genes that may be of importance in malignant progression of papillary thyroid carcinoma (PTC) in the Middle East, which therefore can be targeted in cancer therapy, we screened and validated the global gene expression in PTC using cDNA expression arrays and immunohistochemistry (IHC) on tumour tissue microarrays. Twenty-nine PTC tissue specimens were compared with seven non-cancerous thyroid specimens by use of cDNA microarray. Results for selected genes were confirmed by quantitative real-time PCR. Protein expression of selected genes was further studied using a tissue microarray consisting of 536 PTCs and compared with histologically non-cancerous tissue samples. One hundred and ninety-six genes were overexpressed in PTC tissues relative to non-cancerous thyroid tissues. The genes that were up-regulated in PTC were involved in cell cycle regulation, cell signaling, and oncogenesis. Among these genes, c-MET was identified by immunohistochemical methods as a protein that is overexpressed in 37% of PTCs and was significantly associated with more aggressive behaviour, eg higher stage, nodal involvement, and tall cell variant (p value = 0.01, 0.01 and 0.04, respectively). In this study, 55% of the PTC cases expressed activated AKT (P-AKT), which suggests that activated AKT may play an important role in PTC tumourigenesis. The fact that most of the PTC cases that had activated AKT showed overexpression of c-MET (p = 0.027) leads us to hypothesize that c-MET may be an alternative mechanism of AKT activation in Middle Eastern PTCs. Finally, our data suggest that c-MET dysregulation is associated with aggressive behaviour and may serve as a molecular biomarker and potential therapeutic target in this disease.

The glyoxylate cycle in an arbuscular mycorrhizal fungus. Carbon flux and gene expression.

The arbuscular mycorrhizal (AM) symbiosis is responsible for huge fluxes of photosynthetically fixed carbon from plants to the soil. Lipid, which is the dominant form of stored carbon in the fungal partner and which fuels spore germination, is made by the fungus within the root and is exported to the extraradical mycelium. We tested the hypothesis that the glyoxylate cycle is central to the flow of carbon in the AM symbiosis. The results of (13)C labeling of germinating spores and extraradical mycelium with (13)C(2)-acetate and (13)C(2)-glycerol and analysis by nuclear magnetic resonance spectroscopy indicate that there are very substantial fluxes through the glyoxylate cycle in the fungal partner. Full-length sequences obtained by polymerase chain reaction from a cDNA library from germinating spores of the AM fungus Glomus intraradices showed strong homology to gene sequences for isocitrate lyase and malate synthase from plants and other fungal species. Quantitative real-time polymerase chain reaction measurements show that these genes are expressed at significant levels during the symbiosis. Glyoxysome-like bodies were observed by electron microscopy in fungal structures where the glyoxylate cycle is expected to be active, which is consistent with the presence in both enzyme sequences of motifs associated with glyoxysomal targeting. We also identified among several hundred expressed sequence tags several enzymes of primary metabolism whose expression during spore germination is consistent with previous labeling studies and with fluxes into and out of the glyoxylate cycle.

Chemo- and baroresponses differ in African-Americans and Caucasians in sleep.

To determine sleep effects on baro- and ventilatory responses to transient chemo- and barostimulation in African-Americans and Caucasians, 26 nonobese normotensive young subjects (13 African-Americans and 13 Caucasians) were studied awake and in non-rapid-eye movement (NREM) and rapid-eye-movement sleep during induced transient hypoxemia (N2), hypertension (phenylephrine, PE), and concomitant hypoxemia and hypertension (N2 + PE). Arterial blood pressure was recorded by plethysmographic volume clamp, minute ventilation by pneumotachograph, and arterial O2 saturation by pulse oximeter. For all subjects, chronotropic baroresponse (Deltapulse interval/Deltasystolic blood pressure, where Delta is change) increased with NREM sleep (P = 0.007). Baroresponse slope was greater in Caucasians than in African-Americans (ANOVA, P = 0.02). Hypoxemic ventilatory response (Deltaminute ventilation/Deltaarterial O2 saturation) was greater in African-Americans than in Caucasians in NREM sleep (P = 0.01), as was hypoxemic attenuation of baroresponse (N2 + PE, P = 0.03). These data suggest sleep-related differences in arterial chemo- and baroreceptor responses in normal young African-Americans and Caucasians, which may have implications concerning development of systemic hypertension.

Characteristics of predominantly nonwhite patients with frequent hospitalizations for acute asthma in Chicago.

The purpose of this study was to determine the characteristics of predominantly nonwhite patients with recurrent visits to the emergency department (ED) and admissions to an inner-city hospital in Chicago for acute asthma. Over a 21-month period, two groups of age and gender-matched individuals with asthma seen at the University of Illinois at Chicago Medical Center were studied: group I included 26 patients with frequent visits to the ED and no more than one admission for acute asthma/year; and group II included 28 patients with recurrent visits to the ED and two or more admissions for acute asthma/year. We found that 70% of all patients (38/54) were females and 72% (39/54) were African-Americans. The latter predominated in group II (25/28; 89%). There were no significant differences in public aid recipients, baseline FEV1, type of antiasthma medications used, and illicit drug use between the two groups. However, group II reported more asthma onset before the age of 11 years and used higher daily doses of inhaled corticosteroids than group I (p < 0.05). The average duration of hospital stay in group II was significantly longer (3.3 +/- 0.4 days vs. 2.4 +/- 0.3 days, respectively, mean +/- SEM, p < 0.05), and the average cost per hospitalization in group II significantly exceeded that of group I ($5122 +/- $590 vs. $3740 +/- $450, respectively, p < 0.05). We conclude that African-American females are seen more frequently in the ED for acute asthma and admitted to the hospital in Chicago. They develop asthma before the age of 11 years, use higher daily doses of inhaled corticosteroids, and contribute significantly to the high cost of asthma care.