7). In aggregate, the data suggest that OPN plays a major role in chronic CCl4-induced hepatic fibrosis by regulating scar formation. To confirm the results obtained under chronic CCl4 injection, we used TAA treatment as a second model of chronic drug-induced liver fibrosis. Sirius red/fast green staining selleck chemicals and Collagen-I IHC showed stage >3 fibrosis in TAA-treated WT and ∼1-2 in Opn−/− mice with clear induction of Collagen-I deposition in TAA-treated WT compared to Opn−/−, mice, extensive portal fibrosis, bridging fibrosis and a ∼3-fold increase in scar thickness (Supporting Fig. 8A, 8B). Thus, fibrosis was more distinct in TAA-treated

WT than in Opn−/− mice, as quantified by Brunt fibrosis score and by Sirius red and Collagen-I morphometry (Supporting Fig. 8C-8E). Collectively, these results suggest that increased OPN expression per se or after chronic liver injury and oxidant stress can stimulate Collagen-I deposition in vivo. In addition, the in vitro EPZ-6438 ic50 studies demonstrate that intracellular OPN plays an autocrine role in regulating Collagen-I expression in HSCs. Moreover, treatment with rOPN to resemble the paracrine actions of secreted OPN increases HSC invasion, chemotaxis and wound-healing potential and up-regulates Collagen-I via integrin αvβ3 engagement and activation of PI3K-pAkt-NFκB signaling (Supporting Fig. 9). It is becoming clearer that OPN

is significantly induced during liver injury, both in humans and in rodents.4-6, 17 In the past few years, work from

several groups4-6, 17 studied the potential role of OPN in liver fibrosis, albeit with inconclusive click here results. Studies by Lee et al.5 demonstrated an OPN increase in the culture medium from culture-activated HSCs and under oral CCl4 administration; however, no mechanistic studies were performed to dissect how OPN regulates Collagen-I protein deposition. Lorena et al.6 suggested increased susceptibility to CCl4 injection in Opn−/− mice. The investigators claimed that the protection observed in WT mice was the result of enhancement of hepatocyte survival and reduction in nitric oxide synthase 2 expression; yet, they neither provided IHC for cell-survival markers nor measured the concentration of NO· or ONOO− to support their conclusions and no studies on Collagen-I regulation were performed. Last, a recent publication from Syn et al.4 proposes a role for the Hedgehog-signaling pathway in activating OPN and promoting fibrosis progression in nonalcoholic steatohepatitis; however, it is not clear which OPN isoform the investigators were referring to, and it is Gli1, and not Gli2, expression that is widely considered the most reliable readout for cells undergoing active Hedgehog signaling. Thus, there is a well-timed need for dissecting the molecular mechanism on how this matricellular protein could regulate the fibrogenic response to liver injury and, specifically, Collagen-I protein expression by HSCs.

Materials and Methods: Five naïve HCV pts (GT4d, GT4a, GT4o n=3,1,1) from the Command 4 Study, candidates for pIFN/RBV+DCV treatment were considered. Plasma samples were collected at baseline and during therapy. The presence and frequency of HCV variants within the NS5A quasispecies was analyzed by ultra-deep pyrosequencing (UDPS). Results: Pt1 received pIFN/RBV, Pts2, 3 and 4 pIFN/RBV+DCV; Pt5 was a screening failure. Pt1 was relapser; Pt2 experienced breakthrough at Wk 4; Pts 3 and 4 showed sustained virological response

(SVR), with Epacadostat cell line HCV RNA undetectable since Wk 4. Considering viremic time points for Pts1 and 2, the extent of NS5A diversity was not significantly related to viral GPCR Compound Library cell assay load (r=−0.4, p=0.75 and r=−0.80, p=0.33, respectively). No substitutions were detected at positions related to DCV resistance at T0, with the exception of P58T in Pt3 (SVR). In Pt2 (breakthrough) multiple substitutions at positions 28, 31 and 93, linked to DCV resistance, were observed since Wk 4, with different kinetics (Table). Mutations were frequently associated on the same haplotype (L28S + M31I = 57, 85 and 99% at Wks4, 8 and 9; L28S + M31I + Y93H = 13, 6 and <0.6% at Wks4, 8 and 9). Conclusions: Our data

suggest that GT4d resistance patterns may involve the same amino acid residues described in GT1 and GT4a, although the substitution at position 28 is novel (L28S); UDPS allows establishing the dynamics and early appearance of substitutions potentially associated with antiviral resistance in patients undergoing DCV-based therapy. The dynamics of Y93H in GT4 patients is consistent with previous findings for GT1b patients, showing that it tends to be associated with other mutations; this suggests that this website it may not confer strong selection advantage in DCV-treated patients, as compared to L28S and M31I, which become predominant during virological failure in this study. Dynamics of mutations along treatment in Pt2 (% of quasispecies) Disclosures: Fiona McPhee – Employment: Bristol-Myers Squibb Eric A. Hughes – Employment: Bristol-Myers

Squibb The following people have nothing to disclose: Barbara Bartolini, Raffaella Lionetti, Emanuela Giombini, Chiara Taibi, Marzia Montalbano, Gianpiero D’Offizi, Giuseppe Ippolito, Anna Rosa Garbuglia, Maria R. Capobianchi Background and Aims: The associations between genetic polymorphism in patatin-like phospholipase family 3 protein (PNPLA3) gene and steatosis or fibrosis in chronic hepatitis C have been reported with conflicting results. On the other hand, data regarding the role of PNPLA3 in chronic hepatitis B is scarce. The aim of the present study was to investigate the role of the PNPLA3 genetic polymorphism in chronic hepatitis C and hepatitis B in terms of steatosis, fibrosis and the development of HCC.

27 While ICG-001 expectedly decreased TOPflash reporter activity, it unexpectedly reduced p65 reporter activity, which may be due to an increase in non-CBP-bound pool of β-catenin (Fig. 7D). These findings suggest that β-catenin modulation of NF-κB signaling is regulatable through manipulation of β-catenin expression; however, only agents that suppress total β-catenin levels may be useful to induce p65 activation. We

next examined conditions in which β-catenin was overexpressed both in vitro and in vivo to determine the effect on p65 expression and activity. Hep3B cells were transfected with control plasmid or plasmid expressing constitutively active S33Y/β-catenin or S45Y/β-catenin, simultaneous with p65 or TOPflash GSI-IX datasheet reporters. While expression of mutated β-catenin induced TOPflash activity, it also resulted in significantly repressing A-769662 cost p65 activity (Fig. 7E). We next treated Hep3B cells with an escalating dose of lithium chloride (LiCl), a known inhibitor of GSK-3β that in turn induces β-catenin protein stabilization. This led

to a dose-dependent increase in TOPflash reporter activity and a concomitant and significant decrease in p65 reporter activity (Fig. 7F). Finally, we analyzed human hepatocellular carcinoma (HCC) tissue array via IHC. Tumor-wide glutamine synthetase (GS) staining is a good indicator of β-catenin mutations.28, 29 Of 93 HCC tumors on Biomax HCC tissue array, 30 were GS-positive, consistent with the numbers of HCC with

β-catenin gene mutations (reviewed by Nejak-Bowen check details and Monga9). Of this subset, the majority of GS-positive HCC (63% [19/30]) were negative for p65 (Fig. 8A,B). These findings indicate that β-catenin activation in HCC negatively affects p65 expression and NF-κB activity. β-Catenin is a crucial component of the Wnt pathway, which plays multiple roles in liver homeostasis through its regulation of proliferation, differentiation, and regeneration. However, its role in hepatic injury remains unexplored. The analysis was initiated to test two common modes of hepatocyte apoptosis: Fas- and TNF-α-mediated cell death. We have reported that β-catenin and the HGF receptor c-Met associate at the cell surface.15 c-Met sequestration of the Fas receptor that can prevent Fas-mediated apoptosis in hepatocytes has also been reported.13 We also identified the Fas/β-catenin complex in the liver. Because HGF messenger RNA up-regulation (along with a dramatic reduction in Met protein levels) was evident in KO livers, we hypothesized that basal apoptosis may be due to destabilization of c-Met/Fas/β-catenin complexes, which may enhance free-Fas levels available for engagement with Fas ligands like Jo-2. However, the KO mice were as susceptible as WT mice to Fas-activation.

Male Non-obese Diabetic/Severe Combined Immunodeficiency

(NOD/SCID) mice received abdominal irradiation at a single dose of 5-Gy, and then transfused intravenously with cytokines treated Flk-1+MSCs and monitored body diarrhea, weight and survival for 30-day. Colonization and differentiation of transplanted Flk-1+MSCs in the irradiated intestine were analyzed by histological and immunohistochemaical Cilomilast manufacturer methods. Results: CXCR4 expression in Flk-1+MSCs was up-regulation of by the cytokine cocktail treatment in vitro. The cytokines treated Flk-1+MSCs could significantly extend the life span of the irradiation mice, decrease diarrhea occurrence and improve the small intestinal structural integrity of irradiated mice. Furthermore, GW-572016 the cytokines treated Flk-1+MSCs were migrated and colonized to the small intestine, and differentiated into fibroblastic-like cells. The immunofluorescence staining showed that the transplanted cells could differentiate into vimentin+/α-SMA+ cells. The mechanism may be that cytokines treatment promoted Flk-1+MSCs home to and engraft to injured sites through up-regulation of CXCR4 expression. In addition, transplanted cells may regulate the epithelial stem/progenitor cells directly or indirectly, and modulate the inflammatory response

through the secretion of trophic factors such as SCF, IL-6 and HGF, which facilitating gastrointestinal repair and gut mucosal barrier function. Conclusion: Our study revealed that administration of cytokines treated Flk-1+MSCs might be a novel therapeutic approach for RIGS. Key Word(s): 1. radiation; 2. MSCs; 3. CXCR4; 4. transplantation; Presenting Author: XUDONG TANG Additional Authors: LIYA ZHOU, ZHENHUA LI, SHUTIAN ZHANG, BIN LV, JUNXIANG LI, BAOSHUANG LI, HUIZHEN selleck chemical LI Corresponding Author: XUDONG TANG Affiliations: Xiyuan Hospital affiliated to China Academy of Traditional Chinese Medicine; Peking University Third Hospital; Beijing Friendship Hospital affiliated to Capital Medical University; Zhejiang Hospital of Traditional Chinese Medicine; Dongfang Hospital affiliated to Beijing University of Traditional Chinese Medicine; The No 2.Hospital affiliated

to Tianjin University of Traditional Chinese Medicine Objective: To evaluate the effect and safety of Chinese herbal medicine based on syndrome differentiation and Moluodan on the gastric epithelial dysplasia in patients with chronic atrophic gastritis. Methods: This is a multi-centered, randomized, clinical controlled trial. Patients in western hospitals and traditional Chinese medicine (TCM) hospitals were given Moluodan or herbal medicine, compared with folic acid respectively. Three hundred and eighty-three subjects with atrophic gastritis accompanied with dysplasia were consecutively enrolled from Beijing and Hangzhou in China, 125 patients in Chinese herbal medicine group, 130 in Moluodan group, and 128 in folic acid group.

Male Non-obese Diabetic/Severe Combined Immunodeficiency

(NOD/SCID) mice received abdominal irradiation at a single dose of 5-Gy, and then transfused intravenously with cytokines treated Flk-1+MSCs and monitored body diarrhea, weight and survival for 30-day. Colonization and differentiation of transplanted Flk-1+MSCs in the irradiated intestine were analyzed by histological and immunohistochemaical GSK126 methods. Results: CXCR4 expression in Flk-1+MSCs was up-regulation of by the cytokine cocktail treatment in vitro. The cytokines treated Flk-1+MSCs could significantly extend the life span of the irradiation mice, decrease diarrhea occurrence and improve the small intestinal structural integrity of irradiated mice. Furthermore, Selleckchem Pexidartinib the cytokines treated Flk-1+MSCs were migrated and colonized to the small intestine, and differentiated into fibroblastic-like cells. The immunofluorescence staining showed that the transplanted cells could differentiate into vimentin+/α-SMA+ cells. The mechanism may be that cytokines treatment promoted Flk-1+MSCs home to and engraft to injured sites through up-regulation of CXCR4 expression. In addition, transplanted cells may regulate the epithelial stem/progenitor cells directly or indirectly, and modulate the inflammatory response

through the secretion of trophic factors such as SCF, IL-6 and HGF, which facilitating gastrointestinal repair and gut mucosal barrier function. Conclusion: Our study revealed that administration of cytokines treated Flk-1+MSCs might be a novel therapeutic approach for RIGS. Key Word(s): 1. radiation; 2. MSCs; 3. CXCR4; 4. transplantation; Presenting Author: XUDONG TANG Additional Authors: LIYA ZHOU, ZHENHUA LI, SHUTIAN ZHANG, BIN LV, JUNXIANG LI, BAOSHUANG LI, HUIZHEN selleck chemicals llc LI Corresponding Author: XUDONG TANG Affiliations: Xiyuan Hospital affiliated to China Academy of Traditional Chinese Medicine; Peking University Third Hospital; Beijing Friendship Hospital affiliated to Capital Medical University; Zhejiang Hospital of Traditional Chinese Medicine; Dongfang Hospital affiliated to Beijing University of Traditional Chinese Medicine; The No 2.Hospital affiliated

to Tianjin University of Traditional Chinese Medicine Objective: To evaluate the effect and safety of Chinese herbal medicine based on syndrome differentiation and Moluodan on the gastric epithelial dysplasia in patients with chronic atrophic gastritis. Methods: This is a multi-centered, randomized, clinical controlled trial. Patients in western hospitals and traditional Chinese medicine (TCM) hospitals were given Moluodan or herbal medicine, compared with folic acid respectively. Three hundred and eighty-three subjects with atrophic gastritis accompanied with dysplasia were consecutively enrolled from Beijing and Hangzhou in China, 125 patients in Chinese herbal medicine group, 130 in Moluodan group, and 128 in folic acid group.

, 2003). Frontal-executive functions are known to include the ability to plan ahead and to overcome impulsive behaviour. It would therefore follow that frontal-executive impairments would correlate with the occurrence of relapses. Unfortunately, no studies to date have provided convincing data to support this proposal (Bowden, Crews, Bates, Fals-Stewart, & Ambrose, 2001). A recent study by Loeber et al. (2010) demonstrated a negative effect on cognitive function and recovery in 31 patients. However, they did not show a correlation with the occurrence of relapses, and included participants with a relatively positive prognosis. In contrast, the study presented here examined

only patients with a history of being resistant to therapy, who can therefore be assumed to have a negative prognosis. Our study thus provides preliminary support for a negative buy BVD-523 association between frontal-executive deficits and future prognosis, although

further longitudinal data and replication with larger cohorts AZD2281 are required. A clinical implication may be drawn from these results based on data indicating that cognitive deficits tend to improve with abstinence (Fals-Stewart & Lam, 2010). It may hence be assumed that patients with subtle executive deficits may benefit most from long-term therapeutic options rather than from frequent detoxifications. It is also noteworthy that the cognitive deficits manifested solely in more dedicated neuropsychological tests (TMT and RWT), and would therefore probably have been missed by routine clinical tests. Similarly, while patients did not fulfil the ICD10 criteria for depressive syndrome, they reported significantly more depressive features selleck screening library in the BDI questionnaire

compared with controls. These depressive tendencies may have aggravated executive impairments, but also would not have been detected in routine clinical tests. In summary, the study presented here found that severely alcohol-dependent subjects who have experienced recurrent withdrawals display subtle cognitive deficits. These deficits occurred primarily in the frontal-executive domain, while memory functions and visuospatial capacities were largely spared. Our pilot study therefore suggests that extensive cognitive testing might be a helpful additional tool in assessing therapy-resistant heavy drinkers. Future trials will elucidate the influence these cognitive deficits have on prognosis and quality of life. “
“Impulse control disorders (ICDs) and apathy are recognized as two important neuropsychiatric syndromes associated with Parkinson’s disease (PD), but as yet we understand very little about the cognitive mechanisms underlying them. Here, we review emerging findings, from both human and animal studies, that suggest that impulsivity and apathy are opposite extremes of a dopamine-dependent spectrum of motivated decision making.

35 In summary, we have shown that mouse iPS cells can be induced to efficiently generate intact fetal livers and that hiPS cells can be induced in culture to produce highly differentiated hepatocytes. We acknowledge that compared with the Buparlisib molecular weight in vivo environment of the liver, the conditions in culture are relatively artificial, and this is likely to impact the function of iPS-derived hepatocytes

compared with the native environment. Nevertheless, the data provided above demonstrate the feasibility of generating cells with hepatic characteristics from skin cells through an iPS cell intermediate and that such cells can engraft into the mammalian liver parenchyma. Such proof-of-concept opens up the possibility of producing patient-specific hepatocytes in a relatively simple and straightforward manner with high efficiency.

We are confident that such cells could be immediately useful for the study of hepatocellular disease and basic developmental mechanisms and for drug Saracatinib purchase screening. The authors thank Charles Myers for providing frozen liver samples. Additional Supporting Information may be found in the online version of this article. “
“Hepatocellular carcinoma (HCC) is a highly vascularized tumor with frequent intrahepatic metastasis. Active angiogenesis and metastasis are responsible for rapid recurrence and poor survival of HCC. We previously found that microRNA-29b (miR-29b) down-regulation was significantly associated with poor recurrence-free survival of HCC patients. Therefore, the role of miR-29b in tumor angiogenesis, invasion, and metastasis was further investigated in this study using in vitro check details capillary tube formation and transwell assays, in vivo subcutaneous and orthotopic xenograft mouse models, and Matrigel plug assay, and human HCC samples. Both gain- and loss-of-function studies showed that miR-29b dramatically suppressed

the ability of HCC cells to promote capillary tube formation of endothelial cells and to invade extracellular matrix gel in vitro. Using mouse models, we revealed that tumors derived from miR-29b-expressed HCC cells displayed significant reduction in microvessel density and in intrahepatic metastatic capacity compared with those from the control group. Subsequent investigations revealed that matrix metalloproteinase-2 (MMP-2) was a direct target of miR-29b. The blocking of MMP-2 by neutralizing antibody or RNA interference phenocopied the antiangiogenesis and antiinvasion effects of miR-29b, whereas introduction of MMP-2 antagonized the function of miR-29b. We further disclosed that miR-29b exerted its antiangiogenesis function, at least partly, by suppressing MMP-2 expression in tumor cells and, in turn, impairing vascular endothelial growth factor receptor 2-signaling in endothelial cells. Consistently, in human HCC tissues and mouse xenograft tumors miR-29b level was inversely correlated with MMP-2 expression, as well as tumor angiogenesis, venous invasion, and metastasis.

Methods: During January 2000 and December 2012, 210 stents were placed in 183 patients with malignant esophageal obstruction from esophageal cancer, lung cancer or stomach cancer with esophageal invasion. Dysphagia grade, clinical outcome, complications,

and risk factor of complications were evaluated. Results: The improvement in dysphagia grade after stent implantation was statistically significant. (from 3.2 to 1.8, p < 0.001)Complication occurred in 23 (11%) patients. Procedure-related mortality was 2.4% (5/210). Tumor ingrowth and overgrowth is a significant problem with stent insertion, occurring in 53 patients (29%). And bleeding, sepsis due to procedure is more serious complication in the patients selleck antibody with malignant

dysphagia, and mortality rate is high. When comparing the esophageal, lung, and stomach cancer groups, fistula status (p < 0.001) and migration (p = 0.017) were significantly different from each other. But there were no significant risk factors between complication and non-complication group. Complications were not correlated to type of tumor characteristic (p = 0.176). Conclusion: Expandable metal stents offer excellent palliation Enzalutamide of malignant obstruction. Placement of the expandable metal stents effectively relieved malignant dysphagia in treated patients. Several factors should be considered before applying palliative therapy for malignant esophageal obstruction. Tumor characteristics

such as location, fistula, and type need to be considered. Factors such as medical comorbidity and overall selleck chemicals llc expected duration of survival also are important. Key Word(s): 1. Expandable metal stents; 2. malignant esophageal obstruction Presenting Author: DAIKI MORIKAWA Additional Authors: EIJI HIRAOKA Corresponding Author: DAIKI MORIKAWA Affiliations: Tokyo Bay Urayasu Ichikawa Medical Center Objective: Introduction: Gastric anisakiasis is a parasitic disease caused by the gastric mucosal penetration of the Anisakis larvae ingested with raw marine fish. Anisakis can penetrate small intestinal wall, leading abdominal pain and intestinal obstruction. We reported here a case of gastric anisakiasis with gastric bleeding and a case of small intestinal anisakiasis. Methods: Case1: A 73 year-old Japanese man presented with 1 day history of tarry stool and hematemesis 4 hours after eating a raw mackerel. His vital signs were within normal range. He had no abdominal tenderness. The laboratory findings were not significant. The endoscopy showed A1 stage ulcer and the presence of Anisakis larvae. He was diagnosed with acute gastric anisakiasis with gastric ulcer. It was resolved with proton pump inhibitor and conservative treatment.

[57] Furthermore, as stated by Raddant and Russo,[3] “The inflammatory cascade can ATM inhibitor be triggered by CGRP actions on dural mast cells and satellite glial cells of the trigeminal ganglion. The peripheral CGRP-containing neurons (in the trigeminal ganglion and elsewhere) are polymodal nociceptors that innervate essentially all peripheral tissues and send primary afferent input to the dorsal horn, trigeminal nucleus caudalis, or nucleus of the solitary tract (which, in turn, project to the brainstem, amygdala, hypothalamus, and thalamic nuclei).[48] CGRP-containing neurons in the trigeminal ganglion project to the trigeminal

nucleus caudalis and C1-C2, where CGRP also acts post-junctionally on these second-order neurons to transmit pain signals from the brainstem to the thalamus.[58, 59] The clinical correlation of CGRP actions

at the level of the trigeminal nucleus caudalis is relevant as well. The brainstem has a key role in the pathophysiology Palbociclib cell line of migraine.[60, 61] Brainstem stimulation causes activation of the trigeminovascular system, resulting in peripheral CGRP release and neurogenic inflammation (described earlier).[62, 63] Furthermore, activation of the brainstem is associated with altered perception termed allodynia (a condition in which nonpainful stimulation is perceived as painful) as well as with the development of second- and third-order neuronal sensitization.[64, 65] Accordingly, if we understand migraine as the combined result of altered perception of stimuli that are usually not painful, as well as the activation of see more a feed-forward neurovascular dilator mechanism in the first (ophthalmic) division of the trigeminal nerve, we realize that CGRP is involved in the pathophysiology of migraine both centrally and peripherally.[66] CGRP and its receptors are widely distributed across other parts of the CNS as well, in areas that are relevant to pain and in areas that may not be, such as the cerebellum.[67, 68] The function of CGRP in these areas is not well understood. Studies have suggested that CGRP

is expressed in areas that could explain migraine-related photophobia.[69] In a model of transgenic mice, light-aversive behavior was greatly enhanced by intracerebroventricular injection of CGRP and blocked by coadministration of the CGRP-RA olcegepant.[70] Finally, CGRP seems to be important in determining neuronal plasticity and synapse formation. This is either due to its direct actions on neurons or its indirect actions on the glia via its modulatory actions.71-73 In summary, CGRP and its receptors are largely expressed in neurons and glia, both peripherally and centrally. As discussed later, this broad expression has relevance for drug development. Pain improvement can be achieved by blocking CGRP peripherally, centrally, or both, and brain penetration may not be essential for the analgesic properties of CGRP antagonists.

Queens were isolated with moist paper towels in individual plastic shipping tubes and shipped overnight to the University of Vermont. Upon arrival, queens were individually weighed to the nearest 0.01 mg with a Mettler

Toledo microbalance (AX 205 Microbalance, Mettler-Toledo, Columbus, OH, USA) and painted with one of three different colors of Testors paint pens on the thorax. Pairs of queens differing in paint color and an equal number of single ‘control’ queens were placed into 600-mL bottles 2/3 filled with damp soil in which the queens could excavate a nest and rear brood in a seminatural soil-filled tunnel. Thirty sets of bottles were set up in ABT 263 2011, and 36 in 2012. Division of labor could emerge as a result of multiple types of self-organization

mechanisms (Duarte et al., 2011), including agonistic social interactions (Jeanson et al., 2005). To determine whether agonistic interactions drive division of labor between queens, we quantified the extent and symmetry of aggressive behavior when queens were first introduced. All pairs of queens in both nest types were observed in groups of six nests for the first 15 min following their release into the nest. All instances of aggressive behaviors (Table 1) performed by each queen during this period were recorded. The contribution of each queen to excavation behavior was quantified by intensive observations of groups of 20 nests for 15-min intervals in which all instances of excavation behavior by each queen were noted. A subset of five nests in a set was scanned by a single observer for 3–5 s before moving to the next Daporinad price subset, resulting in approximately two scans per minute per nest over the entire 15-min interval. All observations were conducted over a period of 2 days, after which excavation behavior had ceased and the majority of nests were sealed with soil. In 2011, nests were observed for a total of 10 observation selleck products periods; this was increased to 15 in 2012 to better capture high-intensity excavation bouts in the first few hours following queen introduction. Colonies were

collected in week eight, when the brood in the majority of colonies contained darkening pupae and/or workers. All surviving queens, larvae, pupae and workers were counted and preserved in 95% ethanol. Any pairs in which one or both queens had died prior to collection were excluded from reproduction comparisons. To determine queen lineage identity and reproductive apportionment in paired nests, DNA was extracted from a leg or the head of each queen from both the paired and control nests, and the whole body for all brood from paired nests using a standard Chelex-100 rapid extraction protocol (Helms Cahan et al., 2006). To determine queen lineage identity, the Cox1 mitochondrial gene was amplified as described in Schwander et al.