Search

Smartphones have rapidly become indispensable in modern life. The wide spectrum of activities and virtual interaction possibilities with the mobile device render some individuals prone to exaggerated use exhibiting core symptoms akin to substance addiction. The core common symptoms of addictions can be framed within the ‘components model of addiction.’ Based on the different pattern of smartphone use in various age groups, the symptoms and the risk of smartphone addiction may differ during various life stages. Some age groups may be more at risk of smartphone addiction than others. This hypothesis was tested utilizing data from six surveys comprising 1603 individuals representing various age groups. Results obtained through two recommended age-group analyses both confirmed that preschool children and young adults are at highest risk for smartphone-related addictive behavior. Furthermore, the various components of addiction also differed between the age groups. These results suggest that preventive measures should be primarily targeted to the two most vulnerable age groups represented by preschool children and emerging adults. The results of the present study help identify the dominant addiction symptoms in the various age groups that could pinpoint aspects which should be targeted in implementation of various measures aimed at the prevention of dysfunctional smartphone use.

Significance Standard of care for metastatic castration-resistant prostate cancer (mCRPC) mainly relies on suppression of androgen receptor (AR) signaling. This approach has no lasting benefit due to the emergence of resistance mechanisms, such as ligand-independent splicing variant AR-V7. A metabolic feature of mCRPC is the upregulation of de novo lipogenesis to provide substrates and fuel for metastatic spread. Whether increased levels of fats affect AR signaling to promote an aggressive disease remains to be determined. Using a selective and potent inhibitor of fatty acid synthase we demonstrate that suppression of this key enzyme inhibits AR, most importantly AR-V7, and reduces mCRPC growth. Our findings offer a therapeutic opportunity for mCRPC and a potential mechanism to overcome resistance to AR inhibitors., A hallmark of prostate cancer progression is dysregulation of lipid metabolism via overexpression of fatty acid synthase (FASN), a key enzyme in de novo fatty acid synthesis. Metastatic castration-resistant prostate cancer (mCRPC) develops resistance to inhibitors of androgen receptor (AR) signaling through a variety of mechanisms, including the emergence of the constitutively active AR variant V7 (AR-V7). Here, we developed an FASN inhibitor (IPI-9119) and demonstrated that selective FASN inhibition antagonizes CRPC growth through metabolic reprogramming and results in reduced protein expression and transcriptional activity of both full-length AR (AR-FL) and AR-V7. Activation of the reticulum endoplasmic stress response resulting in reduced protein synthesis was involved in IPI-9119–mediated inhibition of the AR pathway. In vivo, IPI-9119 reduced growth of AR-V7–driven CRPC xenografts and human mCRPC-derived organoids and enhanced the efficacy of enzalutamide in CRPC cells. In human mCRPC, both FASN and AR-FL were detected in 87% of metastases. AR-V7 was found in 39% of bone metastases and consistently coexpressed with FASN. In patients treated with enzalutamide and/or abiraterone FASN/AR-V7 double-positive metastases were found in 77% of cases. These findings provide a compelling rationale for the use of FASN inhibitors in mCRPCs, including those overexpressing AR-V7.

Introduction Symptoms of nicotine dependence among adolescents occur at an early stage in smoking onset and can be present even with low exposure to cigarettes. We aim to examine the early occurrence of symptoms of nicotine dependence and how they predict later smoking behavior. Methods Participants were ninety-four currently smoking 9th-graders attending high school in Targu Mures, Romania. They were followed for 6 months with two assessment points: baseline, and follow-up at 6 months. We assessed the following: 1) the number of smoked cigarettes in the last 30 days, 7 days, and 24 hours using the Minnesota Smoking Index; 2) vulnerability to addiction manifested in cessation difficulties, using the 9-item version of the Hooked On Nicotine Checklist (HONC), 3) loss of autonomy using the endorsement of at least one HONC item, and 4) dependence, using the modified Fagerström Tolerance Questionnaire (mFTQ). We performed statistical analysis with SPSS version 19, using paired-sample t-tests for comparing the differences between baseline and follow-up data. We also conducted linear regression analysis to demonstrate the predictive role of the assessed variables, such as the scores of the mFTQ and the HONC in maintaining smoking and reported smoking status. Results Regression models indicated that baseline-measures for symptoms of dependence (β=0.64, p

Abstract: Introduction and aim: To evaluate the frequency and severity of the malocclusions and treatment need in a group of adult patients in Târgu-Mureș, dividing them up according to age and gender. Method: Sixty-four, randomly selected patients were divided up according to age and gender. Ten occlusal traits related to dentofacial anomalies according to missing teeth, spacing, crowding and occlusion were used to evaluate the malocclusions on each of their study cast. The Dental Aesthetic Index (DAI) regression equation was used to obtain the DAI score that defines severity. Results: The average DAI score was 38 with a range of 18–79, showing that the majority of patients needed mandatory orthodontic treatment. No statistically significant difference was found between gender, age groups and need for treatment. Conclusion: More than two-fifths of the examined patients have a mandatory need for orthodontic treatment if the DAI scores are the main screening tool. Orv Hetil. 2018; 159(9): 352–356.

The role of diet is crucial in the development and prevention of cardiovascular disease. The purpose of the current study was to describe dietary intake and assess health scores of children and adults by using a survey questionnaire on moldavian and transylvanian population, in order to evaluate whether it is necessary to introduce a prevention program for cardiovascular diseases in Romania. In 2015, a questionnaire was sent to 347 children and 583 adults in two counties in Romania where the climatic and topographic conditions are different. Participation rate in the children group was 56.48% moldavian / 43.51% transylvanian, respectively 39.62% moldavian / 60.37% transylvanian participants in the adult group. The questionnaire included a 44-item semi-quantitative food frequency questionnaire and questions on lifestyle for children and a 21- item semi-quantitative food frequency questionnaire and a health score for adults. Results were evaluated against the WHO recommendations of healthy diet. In Mures county the number of former smokers is significantly higher compared to Bacau county, and considerably more subjects report active smoking and exposure to passive smoking. The incidence of overweight and obesity was similar in the two counties, 68.14% in Bacau and 70.67% in Mures. More subjects from Bacau (94.35%) have regular exercise compared to those from Mures (85.38%), the difference is not quite significant. The majority of the subjects from Bacau county have a significantly healthier diet compared to those from Mures, consuming regularly eggs, meat and dairy products. Based on our data, only 16% of Romanian children reach the WHO recommendation for healthy nutrition regarding fruit and vegetable intake.Dietary habits of children and adults in Romania warrant improvements. Public health actions should be taken to increase the consumption of fruits and vegetables, decrease the consumption of saturated fats and sweets and also to increase the frequency of physical activity. These actions may be helpful in reducing the risks for overweight and cardiovascular diseases.

In blood vessels situated just after the heart, an irregular blood flow occurs due to some specific structural elements of the tunica media. The current paper describes the histological aspects of some post-cardiac arterial sections in lamb. The tissue samples were collected from five 30 days old male lambs (Țurcană breed). Histological specimens from different regions of the aorta were harvested (i.e., the ascending aorta, aortic arch, thoracic and abdominal regions of the descending aorta). From the specified regions, small pieces (cca. 0.5 cm) were fixed in neutral 10% buffered formalin. The tissues were subsequently embedded in paraffin wax, sectioned at 5 μm, and stained with Goldner’s trichrome and Verhoeff methods. Tissue analysis was performed using an Olympus system for image acquisition and analysis. Histological appearance of the assessed segments of the aorta in lamb is unusual. Major changes occur in tunica media of the aorta. In the ascending aorta, aortic arch and thoracic regions of the aorta, the histological outline is somewhat the same. The internal region of the media possesses the typical lamellar arrangement. Concerning the outer part of tunica media, the smooth muscle has a tendency to form bundles of various sizes. The muscle islands are not present in the media of abdominal region of the aorta, which exhibits the classic pattern of elastic arteries.

BACKGROUND The role of gamma-synuclein (SNCG) has been widely examined in malignant conditions due to its possible role in disease progression, but very little information is available on its theoretical function on endometriosis formation. MATERIAL AND METHODS Between January 2016 and December 2016, we collected peritoneal fluid and plasma samples from 45 consecutive female patients, of which 15 were without endometriosis, 15 had minimal to mild endometriosis, and 15 had moderate to severe endometriosis. The statistical power was 0.98. We evaluated SNCG levels in the peritoneal fluid and plasma of patients diagnosed with endometriosis, and we compared them with the levels obtained from disease-free control subjects by using enzyme-linked immunosorbent assay. RESULTS SNCG levels were statistically significantly (1.2-fold) higher in the peritoneal fluid of patients with endometriosis compared to controls (p=0.04). We did not find a significant difference between SNCG levels in the plasma of our endometriosis patients and the control group (p=0.086). However, despite previous data showing very limited expression of SNCG in healthy tissues, we found SNCG in the peritoneal fluid of all of the patients in our healthy control group. CONCLUSIONS Levels of SNCG were statistically significantly higher in the peritoneal fluid of patients with endometriosis compared to disease-free controls, which may indicate its possible role the formation and progression of the disease. Moreover, its biological function should be further investigated due to the conflicting results concerning its expression in healthy tissues.

Abstract: Introduction and aim: In the treatment of colorectal endometriosis a multidisciplinary laparoscopic resection is suggested, for this reason the correct selection of bowel infiltration is essential before surgery. Patients and method: Between 2009 and 2015, 383 sigmoidoscopies were performed in patients with endometriosis. Where mucosal invasion was absent secondary signs (wall rigidity, impression, kinking, pain during the examination, suffusion) were analysed. In endoscopically confirmed cases multidisciplinary surgery was performed, the remaining patients were operated by a gynecologic team only. Results: Endometriosis was endoscopically confirmed in 224 patients (58.49%), 108 of them underwent multidisciplinary operation, the negative 135 cases received gynaecological surgery. Bowel endometriosis was confirmed in 103 out of 108 cases intraoperatively, while in 8 cases of the sigmoidoscopically negative patients bowel infiltration was diagnosed intraoperatively by the gynaecological team. Complete sigmoidoscopy was performed in 43.47% of the cases. Intraluminal endometriosis was found in 4.91%, secondary signs as rigidity in 38.39%, impression in 45.54%, kinking in 57.14%, pain (in cases of examination without narcosis) in 26.06% and suffusion in 3.82% of the cases was found during sigmoidoscopy. Sigmoidoscopic examination has a 92.8% specificity and 96.2% sensitivity in cases of bowel endometriosis. Conclusion: Sigmoidoscopy performed by an experienced gastroenterologist is a highly sensitive examination for the diagnosis of bowel endometriosis. Orv. Hetil., 2017, 158(7), 264–269.

Introduction: A number of the patients suffer from endometriosis increased in the past decades and the cases have became more serious. The most critical complication of bowel endometriosis is the large bowel obstruction. Up to recently, 16 similar case reports of large bowel endomteriosis causing obstruction, with detailed medical history have been published in the literature in English language. Patients: Since 2007 535 female have been treated in the 1st Gynaecological Department with endometriosis, out of them three patients from emergency surgery in the history because of large bowel obstruction. Results: Symptoms suggesting endometriosis or previous intervention due to endometriosis were detected in 59% (13/21) of the cases. Preoperative ultrasound, computertomography, magnetic resonance imaging did not give correct diagnosis. Colonoscopy was carried out before the primary operation in 61% (13/21) and after the surgery in 24% (5/21) of the cases, but none of them confirmed endometriosis. Although all the patients developed obstruction, only in 5% (1/19) of the patients was the mucosa infiltrated by the endometriosis. Conclusions: In a young female patient, intestinal obstruction can be caused by bowel endometriosis. Identification of colonoscopic signs (rigidity, impression, kinking) of endometriosis may help to avoid unnecessary extension of intestinal resection. Gynaecologists should take part in the operations. Orv. Hetil., 2016, 157(49), 1960–1966.

STAT3 (signal transducers and activators of transcription 3) is a transcription factor and a member of the STAT protein family that is activated through a variety of different cytokine and growth factor receptors via JAKs, as well as through oncogenic fusion proteins and gain-of-function (GoF) mutations in STAT3 itself. STAT3 hyperactivation and GoF mutations are found in numerous cancers, including clinically aggressive hematologic malignancies with high unmet medical need, such as peripheral T cell lymphomas (PTCLs) (Andersson et al., 2020). We have previously shown that a potent and selective STAT3 heterobifunctional degrader, KTX-201, strongly represses cell growth in models of STAT3-dependent heme malignancies (Csibi et al., 2019). Herein, we report on the cellular mechanisms underlying the anti-tumor effect of STAT3 degradation in PTCL and provide a model for the relationship between pharmacokinetics/ pharmacodynamics (PK/PD) and activity of KTX-201 in vivo. The relationship between STAT3 degradation by KTX-201, anti-tumor mechanism of action and in vivo activity were investigated in anaplastic large T cell lymphoma (ALCL) models, a subset of PTCLs. In vitro, a decrease of STAT3 by 90% for 48hr was required for ALCL cells to commit to death. To identify anti-tumor mechanism(s) of KTX-201 at the systems level, we performed a time-resolved analysis of the proteomic changes of SU-DHL-1 cells undergoing growth inhibition mediated by KTX-201 at GI95. We measured the abundance of 10,000 proteins and confirmed selective degradation of STAT3 by KTX-201 after 8h of treatment. Significant changes in several marker proteins known to be involved in STAT3-mediated proximal signaling in ALCL including SOCS3, Myc and Granzyme B were observed after 16h. Functional annotation analysis of proteins identified pathways that were significantly enriched in at least one time point. Using unsupervised hierarchical clustering of annotations, we found that proteins that increased in abundance over 48h of exposure to KTX-201 were associated with markers of apoptosis and those that decreased in abundance by 24h and 48h were associated with cytokine signaling and cell cycle, respectively. Based on these data, this study identifies inhibition of cytokine signaling, G1 cell cycle arrest and induction of apoptosis as key anti-tumor mechanisms associated with KTX-201 consistent with observed cell phenotypes. STAT3 degradation in tumor was characterized in mice bearing SU-DHL-1 tumors following single dose IV administration. The STAT3 PD response in tumor was correlated with exposures in tumor. At the dose of 25 mg/kg weekly where complete tumor regression was achieved, KTX-201 achieves >90% STAT3 degradation at 24h post dosing in SUDHL1 xenografts. STAT3 degradation was maintained at 90% at 4 days post dosing. The results from the PK/PD study suggests that STAT3 degradation in tumor of >90% is necessary for anti-tumor efficacy in vivo of KTX-201, but only for a limited duration, such as 4 days out of a weekly dosing cycle. Collectively, our data demonstrate that significant STAT3 degradation for a limited time during dosing interval with KTX-201 in ALCL promotes early changes in key signaling nodes involved with proliferation and cytokine stimulation, followed by profound changes in apoptotic proteins. By integrating mechanistic biology with a deep understanding of PK/PD and efficacy, this study provides a foundation for the clinical development of STAT3 degraders using intermittent dosing regimen for treatment of PTCL and other STAT3-dependent heme malignancies. Disclosures Rong: Kymera Therapeutics: Current Employment, Current equity holder in publicly-traded company. Sharma:Kymera Therapeutics: Current Employment, Current equity holder in publicly-traded company. Csibi:Kymera Therapeutics: Current Employment, Current equity holder in publicly-traded company, Ended employment in the past 24 months. Yang:Kymera Therapeutics: Current Employment, Current equity holder in publicly-traded company. Rusin:Kymera Therapeutics: Current Employment, Current equity holder in publicly-traded company. Shi:Kymera Therapeutics: Current Employment, Current equity holder in publicly-traded company. Dey:Kymera Therapeutics: Current Employment, Current equity holder in publicly-traded company. Karnik:Kymera Therapeutics: Current Employment, Current equity holder in publicly-traded company. Mayo:Kymera Therapeutics: Current Employment, Current equity holder in publicly-traded company. Yuan:Kymera Therapeutics: Current Employment, Current equity holder in publicly-traded company. Chutake:Kymera Therapeutics: Current Employment, Current equity holder in publicly-traded company. McDonald:Kymera Therapeutics: Current Employment, Current equity holder in publicly-traded company. Zhu:Kymera Therapeutics: Current Employment, Current equity holder in publicly-traded company. Ji:Kymera Therapeutics: Current equity holder in publicly-traded company, Ended employment in the past 24 months. Liu:Kymera Therapeutics: Current Employment, Current equity holder in publicly-traded company. Li:Kymera Therapeutics: Current Employment, Current equity holder in publicly-traded company. Walker:Kymera Therapeutics: Current Employment, Current equity holder in publicly-traded company. Gollob:Kymera Therapeutics: Current Employment, Current equity holder in publicly-traded company. Mainolfi:Kymera Therapeutics: Current Employment, Current equity holder in publicly-traded company. Desavi:Kymera Therapeutics: Current Employment, Current equity holder in publicly-traded company.

Signal transducer and activator of transcription 3 (STAT3) has been implicated in multiple aspects of tumorigenesis. In addition to increasing cancer cell proliferation and survival, constitutively activated STAT3 is proposed to regulate cross-talk between tumor, stroma and immune cells to promote immune-evasion. STAT3 activity in tumors promotes the production of immune-suppressive factors that activate STAT3 in diverse immune-cell subsets. Mechanistically, genetic studies support a direct role of activated STAT3 in regulating myeloid cell differentiation to contribute to an immune-suppressed tumor microenvironment (TME) (Kortylewski et al.; Nat. Med. 2005 and Curr. Opin. Immunol. 2008) Therefore, STAT3 is a highly attractive target for immune-oncology. Here, we demonstrate that the degradation of STAT3 with a potent and selective STAT3 degrader reverses immune suppression in preclinical models. KYM-003 is a heterobifunctional molecule that hijacks the ubiquitin-proteasome system (UPS) for rapid STAT3 degradation. KYM-003 robustly degraded STAT3 in both human peripheral blood mononuclear cells (hPBMC)-derived monocytes and lymphocytes with DC50 < 100 nM. Degradation in hPBMCs was highly selective for STAT3 vs >10,000 other detected proteins (including all other STAT family members) as evaluated by deep tandem mass tag proteomics. IL-6 treatment of hPBMCs upregulates STAT3 phosphorylation, resulting in transcription of genes involved in myeloid cell-mediated immune suppression, such as IL-10 and CD163, a marker of M2 macrophage differentiation. Treatment of cells with KYM-003 at DC90 for 6 hours abrogated the IL-6 induced up-regulation of immune-suppressive gene signatures consistent with a role of STAT3 in mediating an immune suppressive environment by regulating macrophages and other myeloid cells. In several tumor cell lines, degradation of STAT3 by KYM-003 led to strong downregulation of PD-L1. Importantly, in the BioMap StroNSCLC (DiscoverRx), a co-culture system composed of NCI-H1299 lung cancer cells, hPBMCs and primary human fibroblasts that models immune-suppressed TME biology, KYM-003 treatment decreased angiogenic and immune-suppressive cytokines, including VEGF and IL-10, and promoted the pro-inflammatory anti-tumor cytokines IL-2, IFNg and TNFa. Lastly, administration of KYM-003 to mice bearing syngeneic tumors exhibited anti-tumor activity as monotherapy in the CT26 syngeneic model. We have shown that targeting STAT3 for degradation may have a role in restoring an immune-permissive environment in tumors by both rescuing the suppressed immunologic microenvironment and directly downregulating immune checkpoint signals in tumor cells. These data support STAT3 degraders as a promising new therapeutic modality as immune-oncology agents.Citations: Kortylewski et al (2005) Nat. Med, 11(12):1314-21. Kortylewski et al (2008) Curr. Opin. Immunol, 20(2):228-33 Citation Format: Alfredo Csibi, Bin Yang, Yogesh Chutake, Karen Yuan, Michele Mayo, Veronica Campbell, Alice McDonald, Scott Rusin, Kirti Sharma, Hari Kamadurai, Henry Li, Mike Sintchak, Sean Zhu, Sharon Townson, Anthony Slavin, Haojing Rong, Phillip Liu, Chris De Savi, Jared Gollob, Duncan Walker, Nan Ji, Nello Mainolfi. A STAT3 selective targeted protein degrader decreases the immunesuppressive tumor microenvironment and drives antitumor activity in preclinical models [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr LB-088.

Absztrakt Bevezetés: Laparoscopos vastagbél-resectiónál a specimen transrectalis eltávolításával a hasfali metszés elkerülhető, így az invazivitás tovább csökkenthető. A módszer elfogadottságához előnyeinek és hátrányainak értékelése mellett a technika pontos kidolgozása szükséges. Cikkünk célja az általunk kidolgozott technika és első eredményeink ismertetése. Betegek és módszer: Multidiszciplináris együttműködés keretében 2014. április 16. és 2015. november 1. között 45 laparoscopos bélresectio történt belet infiltráló endometriosis miatt, amelynek során az operált betegek közül 11-nél a speciment transrectalisan távolítottuk el. Az eltávolítandó bélszakasz két végének lekötése és a resectiótól proximalis bélszakasz laparoscopos bulldoggal történő lezárása után átvágtuk a belet, majd a speciment a transrectalisan bevezetett kamerazsákon keresztül távolítottuk el. A proximalis bélvégbe laparoscoposan dohányzacskóöltést helyeztünk be, amelybe a transrectalisan bejuttatott körvarrógép fejét helyeztük. A rectumcsonk varrógéppel történő lezárása után körvarró géppel készítettük el az anastomosist. A módszert a rectum felső harmadát, illetve a szigmabelet érintő endometriosis esetén alkalmaztuk. Eredmények: A műtéti idő a transabdominalis és transrectalis specimeneltávolítás során nem különbözött szignifikánsan (108 perc vs. 118 perc). Az operált betegeknél nem volt szignifikáns különbség a postoperativ első és második napon a fehérvérsejtszámban, illetve nem észleltünk varratelégtelenséget. Következtetés: A munkacsoport által kidolgozott technikával az infekció minimálisra csökkenthető. A transrectalis specimeneltávolítás nem növeli a szövődmények esélyét, és biztonsággal alkalmazható a specimen eltávolításához bélresectio esetén.

Introduction: Traditional surgeries performed in cases of deep infiltrating endometriosis lead to impaired quality of life. Aim: To summarize the postoperative outcome and to compare the rate of postoperative complications after different therapeutic approaches applied in deep infiltrating endometriosis. Method: The authors analized the articles published between March 31, 2004 and March 31, 2015, in the database http://www.pubmed.org using the following keywords: endometriosis, deep infiltrating, nerve sparing, surgery. Results: Non-nerve sparing surgery resulted in temporary urinary dysfunction in 19.1–38.5% of patients, while it occurred in 0.61–33.3% of patients after nerve-sparing surgery. Non-nerve sparing surgical technique resulted in an avarage of 121 days of need for self-catheretisation. When nerve-sparing surgeries were performed the duration of self-catheterisation varied between 7 to 39.8 days. After nerve sparing surgeries, permanent bladder dysfunction was not detected in any case. Conclusions: Because of the successful treatment of the patients symptoms and the lower postoperative complication rate, nerve-sparing surgical technique leads to a significant improvement of the quality of life. Orv. Hetil., 2015, 156(48), 1960–1965.

The goal of our study is to identify possible sources of aggressivity, such as the self-appreciation and modalities of coping used by the teenage participants. Our analysis refers to gender, self-i...

Introduction. Smoking is an important public health issue nowadays. It causes a lot of diseases and represents also a source of carcinogenic substances. Recent studies showed an increased incidence of colorectal cancer in smokers. The aim of our study is to assess the association between smoking and colorectal cancer and to establish the prevalence of heavy smokers among the patients operated on for colorectal cancer. Methodology. We run a retrospective study of the charts belonging to the patients diagnosed with colorectal cancer and operated on in our department between 2004 and 2013. The patients were classified in smokers, former smokers and nonsmokers. The amount of tobacco was evaluated according to the number of smoked cigarettes per day, the smoking period, respectively the pack-years. The data were corroborated with the location of the tumor and analyzed using the online version of Graphpad. Results. From 982 patients diagnosed with colorectal cancer, we found 297 smokers (30.24%). Among these, 106 patients (35.69%) have smoked for over 30 years, at least 20 cigarettes per day, more than 30 pack-years. The number of heavy smokers was significantly greater (p=0.0001) in the group with rectal cancer compared to the group with colon cancer. The association of smoking with rectal cancer was also important (p=0.0015) among the former smokers. Conclusions. Smoking is related to higher incidence of colorectal cancer. Our data sustain the hypothesis of increased risk of developing rectal cancer in heavy smokers. We recommend the screening for colorectal cancer among the heavy smoker population.

STUDY OBJECTIVE We first present a detailed description of a modified NOSE-colectomy technique. Secondly, we report the postoperative outcomes of our prospective case series when compared to conventional laparoscopic bowel resection in a relatively large series of patients. DESIGN Canadian Task Force Classification II-1. SETTING University tertiary referral center. PATIENTS Patient selection The last 90 consecutive patients in our care with DIE of the bowel are presented in this study. Patients were diagnosed at the 1Department of OB/GYN Semmelweis Universtiy Budapest, Hungary. INTERVENTIONS We performed laparoscopic bowel resection using the transrectal NOSE-technique and compared the results of the new operative method (n=30) to traditional laparoscopic bowel resection (n=60). MEASUREMENTS AND MAIN RESULTS Duration of operations The median duration of surgery was 121 minutes in the control group and 96 minutes in the NOSE-group (p=.005). Postoperative complications According to Clavien-Dindo classification, we observed a severe, grade IIIb or higher, overall complication rate of 3.3% among all 90 patients. In the control group anastomosis insufficiency occurred in 3.3% of patients (2/60 cases) and in one patient with anastomotic leakage a rectovaginal fistula was observed (1.7%). There was no significant difference in rates of severe postoperative complications (p=0.55). Hospital stay The length of hospital stay in the control group was a median of 7 (5-13) days whereas in the NOSE-group this was 6 (3-11) days (p

Targeted protein degradation mediated by small molecule degraders represents a new and exciting therapeutic modality to target difficult-to-drug oncogenic proteins including transcription factors. These molecules bind to both the target protein and an E3 ligase, enabling the formation of a ternary complex that leads to ubiquitination and subsequent degradation of the target protein by the proteasome. STAT3 (signal transducers and activators of transcription 3) is a transcription factor and a member of the STAT protein family. In response to cytokines and growth factors, STAT3 is phosphorylated by receptor-associated serine/threonine kinases, and phosphylated STAT3 (pSTAT3) then forms dimers that translocate into the nucleus, binds to DNA, and regulate transcription. STAT3 is frequently mutated and activated in numerous cancers including clinically aggressive hematologic malignancies with high unmet medical need. Mechanistically, aberrant activation of STAT3 has been directly linked to the promotion of cancer cell survival, proliferation and immune evasion, thus making it a highly attractive target for oncology. Potent and selective agents specifically and directly targeting STAT3 have remained elusive, however. Herein we report the discovery of a potent and selective STAT3 heterobifunctional degrader, KYM-003, which displays strong anti-tumor activity in models of STAT3-dependent heme malignancies. KYM-003 degrades STAT3 via an E3 ligase-dependent mechanism. It strongly binds to STAT3 and a E3 ligase, leading to the formation of the a productive ternary complex, which leads to ubiquitination of STAT3 and subsequent proteasomal degradation. KYM-003 robustly degraded STAT3 in a number of primary cells or cell lines with DC50 < 100 nM. Degradation was highly selective for STAT3 vs >10,000 other detected proteins (including all other STAT family members) in cell lines and human PBMCs. Degradation of STAT3 by KYM-003 led to significant downregulation of STAT3 target genes, such as SOCS3, MYC, and PIM1. Importantly, total STAT3 and pSTAT3 levels in tumors were reduced by >90% for at least 24 hours after a single dose of KYM-003 and repeated dosing of KYM-003 showed dose-dependent antitumor activity in xenograft models of heme malignancies. Collectively, our data demonstrates that KYM-003 is a potent and selective STAT3 degrader that exhibited strong anti-tumor activity in vitro and in vivo. These data support STAT3 degraders as a new and exciting therapeutic opportunity in heme malignancies. Disclosures Csibi: kymera Therapeutics: Employment, Equity Ownership. Ji:Kymera Therapeutics: Employment, Equity Ownership. Yang:Kymera Therapeutics: Employment, Equity Ownership. Yuan:Kymera Therapeutics: Employment, Equity Ownership. Mayo:kymera Therapeutics: Employment, Equity Ownership. Rong:Kymera Therapeutics: Employment, Equity Ownership. Rusin:Kymera Therapeutics: Employment, Equity Ownership. Sharma:kymera Therapeutics: Employment, Equity Ownership. Loh:Kymera Therapeutics: Employment, Equity Ownership. Li:Kymera Therapeutics: Employment, Equity Ownership. Townson:Kymera Therapeutics: Employment, Equity Ownership. Chen:kymera therapeutics: Employment, Equity Ownership. Kamadurai:Kymera Therapeutics: Employment, Equity Ownership. Walker:Kymera Therapeutics: Employment, Equity Ownership. Gollob:Kymera Therapeutics: Employment, Equity Ownership. Mainolfi:Kymera Therapeutics: Employment, Equity Ownership.

Estradiol enhances COX-2 expression and prostaglandin biosynthesis in TSC2-deficient cells via a rapamycin-insensitive, mTORC2-dependent mechanism., Lymphangioleiomyomatosis (LAM) is a progressive neoplastic disorder that leads to lung destruction and respiratory failure primarily in women. LAM is typically caused by tuberous sclerosis complex 2 (TSC2) mutations resulting in mTORC1 activation in proliferative smooth muscle–like cells in the lung. The female predominance of LAM suggests that estradiol contributes to disease development. Metabolomic profiling identified an estradiol-enhanced prostaglandin biosynthesis signature in Tsc2-deficient (TSC−) cells, both in vitro and in vivo. Estradiol increased the expression of cyclooxygenase-2 (COX-2), a rate-limiting enzyme in prostaglandin biosynthesis, which was also increased at baseline in TSC-deficient cells and was not affected by rapamycin treatment. However, both Torin 1 treatment and Rictor knockdown led to reduced COX-2 expression and phospho-Akt-S473. Prostaglandin production was also increased in TSC-deficient cells. In preclinical models, both Celecoxib and aspirin reduced tumor development. LAM patients had significantly higher serum prostaglandin levels than healthy women. 15-epi-lipoxin-A4 was identified in exhaled breath condensate from LAM subjects and was increased by aspirin treatment, indicative of functional COX-2 expression in the LAM airway. In vitro, 15-epi-lipoxin-A4 reduced the proliferation of LAM patient–derived cells in a dose-dependent manner. Targeting COX-2 and prostaglandin pathways may have therapeutic value in LAM and TSC-related diseases, and possibly in other conditions associated with mTOR hyperactivation.

Genomic analyses of cancer have identified recurrent point mutations in the RNA splicing factor–encoding genes SF3B1, U2AF1, and SRSF2 that confer an alteration of function(1–6). Cancer cells bearing these mutations are preferentially dependent on wild-type (WT) spliceosome function(7–11), but clinically relevant means to therapeutically target the spliceosome do not currently exist. Here we describe an orally available modulator of the SF3b complex, H3B-8800, which potently and preferentially kills spliceosome-mutant epithelial and hematologic tumor cells. These killing effects of H3B-8800 are due to its direct interaction with the SF3b complex, as evidenced by loss of H3B-8800 activity in drug-resistant cells bearing mutations in genes encoding SF3b components. Although H3B-8800 modulates WT and mutant spliceosome activity, the preferential killing of spliceosome-mutant cells is due to retention of short, GC-rich introns, which are enriched for genes encoding spliceosome components. These data demonstrate the therapeutic potential of splicing modulation in spliceosome-mutant cancers.

Introduction Although web-based, multimedia smoking prevention programs have been tested in several high-income countries, their efficacy in Central and Eastern Europe is unknown. The aim of this trial was to assess the short-term effects of ASPIRA, among Romanian and Hungarian speaking ninth graders in Tirgu Mures, Romania. ASPIRA is the Romanian acronym for the translated and adapted version of ASPIRE, "A Smoking Prevention Interactive Experience," an evidence-based smoking prevention program originally developed to prevent tobacco use among high school students in the United States. Methods Sixteen high schools in Tirgu Mures, Romania were randomized to receive five weekly sessions of the ASPIRA web-based, multimedia program or to a control condition. Socio-demographic data, psychosocial characteristics, and smoking behavior were collected from students at baseline and at 6 months. A hierarchical logistic regression analysis was conducted to test the efficacy of the intervention on smoking initiation and current smoking among 1369 students. Results Never-smoker students in the intervention arm were 35% less likely to report smoking initiation 6 months after the baseline assessment (OR = 0.65, 95%CI: 0.44-0.97). Reduced smoking initiation was observed most notably among students who were exposed to at least 75% of the ASPIRA program. There was no statistically significant effect of the intervention on current tobacco use (OR = 0.80, 95%CI: 0.44-1.46). Conclusions ASPIRA, an adapted version of the evidence-based, multimedia ASPIRE program that was originally developed and tested in the United States may decrease smoking initiation among multi-ethnic adolescents in Central and Eastern Europe. Implications (1). Web-based, multimedia smoking prevention programs may be effective tools to prevent smoking initiation among multi-ethnic adolescent communities in Central and Eastern Europe. (2). The degree of exposure is critical, only high exposure to the multimedia smoking prevention program is associated with reduced smoking initiation.

International audience; The AMPactivated protein kinase (AMPK) is a serine/threonine protein kinase that acts as a sensor of cellular energy status switch regulating several systems including glucose and lipid metabolism. Recently, AMPK has been implicated in the control of skeletal muscle mass by decreasing mTORC1 activity and increasing protein degradation through regulation of ubiquitin-proteasome and autophagy pathways. In this review, we give an overview of the central role of AMPK in the control of skeletal muscle plasticity. We detail particularly its implication in the control of the hypertrophic and atrophic signaling pathways. In the light of these cumulative and attractive results, AMPK appears as a key player in regulating muscle homeostasis and the modulation of its activity may constitute a therapeutic potential in treating muscle wasting syndromes in humans.

The control of muscle cell size is a physiological process balanced by a fine tuning between protein synthesis and protein degradation. MAFbx/Atrogin-1 is a muscle specific E3 ubiquitin ligase upregulated during disuse, immobilization and fasting or systemic diseases such as diabetes, cancer, AIDS and renal failure. This response is necessary to induce a rapid and functional atrophy. To date, the targets of MAFbx/Atrogin-1 in skeletal muscle remain to be identified. We have recently presented evidence that eIF3-f, a regulatory subunit of the eukaryotic translation factor eIF3 is a key target that accounts for MAFbx/Atrogin-1 function in muscle atrophy. More importantly, we showed that eIF3-f acts as a "translational enhancer" that increases the efficiency of the structural muscle proteins synthesis leading to both in vitro and in vivo muscle hypertrophy. We propose that eIF3-f subunit, a mTOR/S6K1 scaffolding protein in the IGF-1/Akt/mTOR dependent control of protein translation, is a positive actor essential to the translation of specific mRNAs probably implicated in muscle hypertrophy. The central role of eIF3-f in both the atrophic and hypertrophic pathways will be discussed in the light of its promising potential in muscle wasting therapy.

International audience; In response to cancer, AIDS, sepsis and other systemic diseases inducing muscle atrophy, the E3 ubiquitin ligase Atrogin1/MAFbx (MAFbx) is dramatically upregulated and this response is necessary for rapid atrophy. However, the precise function of MAFbx in muscle wasting has been questioned. Here, we present evidence that during muscle atrophy MAFbx targets the eukaryotic initiation factor 3 subunit 5 (eIF3-f) for ubiquitination and degradation by the proteasome. Ectopic expression of MAFbx in myotubes induces atrophy and degradation of eIF3-f. Conversely, blockade of MAFbx expression by small hairpin RNA interference prevents eIF3-f degradation in myotubes undergoing atrophy. Furthermore, genetic activation of eIF3-f is sufficient to cause hypertrophy and to block atrophy in myotubes, whereas genetic blockade of eIF3-f expression induces atrophy in myotubes. Finally, eIF3-f induces increasing expression of muscle structural proteins and hypertrophy in both myotubes and mouse skeletal muscle. We conclude that eIF3-f is a key target that accounts for MAFbx function during muscle atrophy and has a major role in skeletal muscle hypertrophy. Thus, eIF3-f seems to be an attractive therapeutic target

It is well known that the large caliber vessels, the nutrition of the wall differs in the internal part in comparison to the external one. In the present study we assessed the presence and distribution of vascularisation of aortic media in 3 species: lamb, rabbit and chinchilla. Samples were harvested from a group of 3 animals for each species. After histological processing, we stained the sections and assessed the presence and distribution of vasa vasorum. In lamb, blood vessels were present in the media of the first aortic segments (with a larger caliber), and starting with abdominal descending aorta they were absent. In rabbit and chinchilla, blood vessels were not present in tunica media. The present study shows that vasa vasorum are present only in tunica media of arteries exceeding a certain caliber.

Tunica adventitia of large caliber vessels in lambs presents a highly different thickness from one segment to another, depending on its position in relation to the heart and the irrigated area. In the case of all the studied arteries, adventitia is formed out of collagen and elastic fibers. In some arteries, adventitia contains a small quantity of elastic fibers (ascending aorta, aortic arch, thoracic descending aorta, bicarotid trunk, left subclavian artery). Other arteries contain elastic fibers in a markedly larger quantity, without exceeding a certain limit (abdominal descending aorta, left and right internal iliac arteries, brachiocephalic trunk, right subclavian artery). There are also vessels in which the elastic component from tunica adventitia is formed out of thick and numerous fibers, so that the elastic component in adventitia markedly exceeds the one from tunica media (left and right external iliac arteries, left and right carotid arteries). The large quantity of elastic tissue in tunica adventitia of these vessels seems to be an adaptation to the external solicitations that exert upon them, in direct relation to the irrigated anatomical area.

The stratigraphic dissection of the thorax and ventral cervical region was made on 5 rabbit cadavers. We firstly exposed the heart, and then the aortic arch. The aortic arch has a very curved trajectory and two collaterals (brachiocephalic trunk and left subclavian artery). The brachiocephalic trunk bifurcates in two collaterals (left and right common carotid arteries) and it ends with the right common carotid artery. The bicarotid trunk is absent in leporidae. In subjects taken into study, the diameter of the right subclavian artery (collateral of the brachiocephalic trunk) is larger than the one in the right common carotid artery (terminal of the brachiocephalic trunk).

Genomic characterization of hematologic and solid cancers has revealed recurrent somatic mutations affecting genes encoding the RNA splicing factors SF3B1, U2AF1, SRSF2 and ZRSR2. Recent data reveal that these mutations confer an alteration of function inducing aberrant splicing and rendering spliceosome mutant cells preferentially sensitive to splicing modulation compared with wildtype (WT) cells. Here we describe a novel orally bioavailable small molecule SF3B1 modulator identified through a medicinal chemistry effort aimed at optimizing compounds for preferential lethality in spliceosome mutant cells. H3B-8800 potently binds to WT or mutant SF3b complexes and modulates splicing in in vitro biochemical splicing assays and cellular pharmacodynamic assays. The selectivity of H3B-8800 was confirmed by observing lack of activity in cells expressing SF3B1R1074H, the SF3B1 mutation previously shown to confer resistance to other splicing modulators. Although H3B-8800 binds both WT and mutant SF3B1, it results in preferential lethality of cancer cells expressing SF3B1K700E, SRSF2P95H, or U2AF1S34F mutations compared to WT cells. In animals xenografted with SF3B1K700E knock-in leukemia K562 cells or mice transplanted with Srsf2P95H/MLL-AF9 mouse AML cells, oral H3B-8800 treatment demonstrated splicing modulation and inhibited tumor growth, while no therapeutic impact was seen in WT controls. These data were also evident in patient-derived xenografts (PDX) from patients with CMML where H3B-8800 resulted in a substantial reduction of leukemic burden only in SRSF2-mutant but not in WT CMML PDX models. Additionally, due to the high frequency of U2AF1 mutations in non-small cell lung cancer, H3B-8800 was tested in U2AF1S34F-mutant H441 lung cancer cells. Similar to the results from leukemia models, H3B-8800 demonstrated preferential lethality of U2AF1-mutant cells in vitro and in in vivo orthotopic xenografts at well tolerated doses. RNA-seq of isogenic K562 cells treated with H3B-8800 revealed dose-dependent inhibition of splicing. Although global inhibition of RNA splicing was not observed; H3B-8800 treatment led to preferential intron retention of transcripts with shorter and more GC-rich regions compared to those unaffected by drug. Interestingly, H3B-8800-retained introns commonly disrupted the expression of spliceosomal genes, suggesting that the preferential effect of H3B-8800 on spliceosome mutant cells is due to the dependency of these cells on expression of WT spliceosomal genes. These data identify a novel therapeutic approach with selective lethality in leukemias and lung cancers bearing a spliceosome mutation. Despite the essential nature of splicing, cancer cells without a spliceosome mutation were less sensitive to H3B-8800 compared with potent eradication of mutant counterparts. H3B-8800 is currently undergoing clinical evaluation in patients with MDS, AML, and CMML. Citation Format: Silvia Buonamici, Akihide Yoshimi, Michael Thomas, Michael Seiler, Betty Chan, Benjamin Caleb, Fred Csibi, Rachel Darman, Peter Fekkes, Craig Karr, Gregg Keaney, Amy Kim, Virginia Klimek, Pavan Kumar, Kaiko Kunii, Stanley Chun-Wei Lee, Xiang Liu, Crystal MacKenzie, Carol Meeske, Yoshiharu Mizui, Eric Padron, Eunice Park, Ermira Pazolli, Sudeep Prajapati, Nathalie Rioux, Justin Taylor, John Wang, Markus Warmuth, Huilan Yao, Lihua Yu, Ping Zhu, Omar Abdel-Wahab, Peter Smith. H3B-8800, a novel orally available SF3b modulator, shows preclinical efficacy across spliceosome mutant cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1185. doi:10.1158/1538-7445.AM2017-1185

Large arteries from 2 rabbits were processed in order to make histological investigations, using two staining techniques: Goldner’s Trichrome and Verhoeff. We noticed that the proportion of the three arterial tunicae modifies as they get farther away from the heart. The changes appear in tunica media, which gradually decreases, and tunica adventitia, which gradually increases. Structurally, we observed that the first arterial segments are typical elastic arteries (pulmonary trunk, ascending aorts, aortic arch and thoracic descending aorta), the next are transitional arteries (abdominal descending aorta, subclavian arteries, carotids), and the iliac arteries are typical muscular arteries.

Polycystic ovary syndrome (PCOS) causes vascular damage to arteries; however, there are no data for its effect on veins. Our aim was to clarify the effects of dihydrotestosterone (DHT)-induced PCOS both on venous biomechanics and on pharmacological reactivity in a rat model and to test the possible modulatory role of vitamin D3(vitD). PCOS was induced in female Wistar rats by DHT treatment (83 μg/day, subcutaneous pellet). After 10 wk, the venous biomechanics, norepinephrine (NE)-induced contractility, and acetylcholine-induced relaxation were tested in saphenous veins from control animals and from animals treated with DHT or DHT with vitD using pressure angiography. Additionally, the expression levels of endothelial nitric oxide synthase (eNOS) and cyclooxygenase (COX-2) were measured using immunohistochemistry. Increased diameter, wall thickness, and distensibility as well as decreased vasoconstriction were detected after the DHT treatment. Concomitant vitD treatment lowered the mechanical load on the veins, reduced distensibility, and resulted in vessels that were more relaxed. Although there was no difference in the endothelial dilation tested using acetylcholine (ACh), the blocking effect of NG-nitro-l-arginine methyl ester (l-NAME) was lower and was accompanied by lower COX-2 expression in the endothelium after the DHT treatment. Supplementation with vitD prevented these alterations. eNOS expression did not differ among the three groups. We conclude that the hyperandrogenic state resulted in thicker vein walls. These veins showed early remodeling and altered vasorelaxant mechanisms similar to those of varicose veins. Alterations caused by the chronic DHT treatment were prevented partially by concomitant vitD administration.

Metformin inhibits cancer cell proliferation, and epidemiology studies suggest an association with increased survival in patients with cancer taking metformin; however, the mechanism by which metformin improves cancer outcomes remains controversial. To explore how metformin might directly affect cancer cells, we analyzed how metformin altered the metabolism of prostate cancer cells and tumors. We found that metformin decreased glucose oxidation and increased dependency on reductive glutamine metabolism in both cancer cell lines and in a mouse model of prostate cancer. Inhibition of glutamine anaplerosis in the presence of metformin further attenuated proliferation, whereas increasing glutamine metabolism rescued the proliferative defect induced by metformin. These data suggest that interfering with glutamine may synergize with metformin to improve outcomes in patients with prostate cancer., German Science Foundation (Grant FE1185), National Institutes of Health (U.S.), Glenn Foundation for Medical Research, National Institutes of Health (U.S.) (Grant 5-P50-090381-09), National Institutes of Health (U.S.) (Grant 5-P30-CA14051-39), Burroughs Wellcome Fund, Smith Family Foundation, Damon Runyon Cancer Research Foundation, National Institutes of Health (U.S.) (Grant 1R01DK075850-01), National Institutes of Health (U.S.) (Grant 1R01CA160458-01A1)

The eukaryotic initiation factor 3 subunit f (eIF3f) is one of the 13 subunits of the translation initiation factor complex eIF3 required for several steps in the initiation of mRNA translation. In skeletal muscle, recent studies have demonstrated that eIF3f plays a central role in skeletal muscle size maintenance. Accordingly, eIF3f overexpression results in hypertrophy through modulation of protein synthesis via the mTORC1 pathway. Importantly, eIF3f Was described as a target of the E3 ubiquitin ligase MAFbx/atrogin-1 for proteasome-mediated breakdown under atrophic conditions. The biological importance of the MAFbx/atrogin-1-dependent targeting of eFI3f is highlighted by the finding that expression of ad eIF3f mutant insensitive to MAFbx/atrogin-1 polyubiquitination is associated with enhanced protection against starvation-induced muscle atrophy. A better understanding of the precise role of this subunit should lead to the development of new therapeutic approaches to prevent or limit muscle wasting that prevails in numerous physiological and pathological states such as immobilization, aging, denervated conditions, neuromuscular diseases, AIDS, cancer, diabetes. This article is part of a Directed Issue entitled: Molecular basis of muscle wasting. (C) 2013 Elsevier Ltd. All rights reserved.

Aims In polycystic ovary syndrome (PCOS), metabolic and cardiovascular dysfunction is related to hyperandrogenic status and insulin resistance, however, Vitamin D3 has a beneficial effect partly due to its anti-oxidant capacity. Nitrative stress is a major factor in the development of cardiovascular dysfunction and insulin resistance in various diseases. Our aim was to determine the effects of vitamin D3 in a rat model of PCOS, particularly the pathogenic role of nitrative stress. Main methods Female Wistar rats weighing 100–140 g were administered vehicle (C), dihydrotestosterone (DHT) or dihydrotestosterone plus vitamin D3 (DHT + D) (n = 10 per group). On the 10th week, acetylcholine (Ach) induced relaxation ability of the isolated thoracic aorta rings was determined. In order to examine the possible role of endothelial nitric oxide synthase (eNOS) and cyclooxygenase-2 (COX-2) pathways in the impaired endothelial function, immunohistochemical labeling of aortas with anti-eNOS and anti-COX-2 antibodies was performed. Leukocyte smears, aorta and ovary tissue sections were also immunostained with anti-nitrotyrosine antibody to determine nitrative stress. Key findings Relaxation ability of aorta was reduced in group DHT, and vitamin D3 partly restored Ach induced relaxation. eNOS labeling was significantly lower in DHT rats compared to the other two groups, however COX-2 staining showed an increment. Nitrative stress showed a significant increase in response to dihydrotestosterone, while vitamin D3 treatment, in case of the ovaries, was able to reverse this effect. Significance Nitrative stress may play a role in the pathogenesis of PCOS and in the development of the therapeutic effect of vitamin D3.

Correspondence to: Anthony MJ Sanchez, e-mail: anthony.mj.sanchez@hotmail.fr; International audience; In skeletal muscle, protein levels are determined by relative rates of protein synthesis and breakdown. The balance between synthesis and degradation of intracellular components determines the overall muscle fiber size. AMP-activated protein kinase (AMPK), a sensor of cellular energy status, was recently shown to increase myofibrillar protein degradation through the expression of MAFbx and MuRF1. In the present study, the effect of AMPK activation by AICAR on autophagy was investigated in muscle cells. Our results show that FoxO3a transcription factor activation by AMPK induces the expression of the autophagy-related proteins LC3B-II, Gabarapl1, and Beclin1 in primary mouse skeletal muscle myotubes and in the Tibialis anterior (TA) muscle. Time course studies reveal that AMPK activation by AICAR leads to a transient nuclear relocalization of FoxO3a followed by an increase of its cytosolic level. Moreover, AMPK activation leads to the inhibition of mTORC1 and its subsequent dissociation of Ulk1, Atg13, and FIP200 complex. Interestingly, we identify Ulk1 as a new interacting partner of AMPK in muscle cells and we show that Ulk1 is associated with AMPK under normal conditions and dissociates from AMPK during autophagy process. Moreover, we find that AMPK phosphorylates FoxO3a and Ulk1. In conclusion, our data show that AMPK activation stimulates autophagy in skeletal muscle cells through its effects on the transcriptional function of FoxO3a and takes part in the initiation of autophagosome formation by interacting with Ulk1. Here, we present new evidences that AMPK plays a crucial role in the fine tuning of protein expression programs that control skeletal muscle mass.

The elevated metabolic requirements of cancer cells reflect their rapid growth and proliferation and are met through mutations in oncogenes and tumor suppressor genes that reprogram cellular processes. For example, in tuberous sclerosis complex (TSC)-related tumors, the loss of TSC1/2 function causes constitutive mTORC1 activity, which stimulates glycolysis, resulting in glucose addiction in vitro. In research published in Cell and Bioscience, Jiang and colleagues show that pharmacological restriction of glucose metabolism decreases tumor progression in a TSC xenograft model. See research article: http://www.cellandbioscience.com/content/1/1/34

International audience; Ubiquitin ligase Atrogin1/Muscle Atrophy F-box (MAFbx) up-regulation is required for skeletal muscle atrophy but substrates and function during the atrophic process are poorly known. The transcription factor MyoD controls myogenic stem cell function and differentiation, and seems necessary to maintain the differentiated phenotype of adult fast skeletal muscle fibres. We previously showed that MAFbx mediates MyoD proteolysis in vitro. Here we present evidence that MAFbx targets MyoD for degradation in several models of skeletal muscle atrophy. In cultured myotubes undergoing atrophy, MAFbx expression increases, leading to a cytoplasmic-nuclear shuttling of MAFbx and a selective suppression of MyoD. Conversely, transfection of myotubes with sh-RNA-mediated MAFbx gene silencing (shRNAi) inhibited MyoD proteolysis linked to atrophy. Furthermore, overexpression of a mutant MyoDK133R lacking MAFbx-mediated ubiquitination prevents atrophy of mouse primary myotubes and skeletal muscle fibres in vivo. Regarding the complex role of MyoD in adult skeletal muscle plasticity and homeostasis, its rapid suppression by MAFbx seems to be a major event leading to skeletal muscle wasting. Our results point out MyoD as the second MAFbx skeletal muscle target by which powerful therapies could be developed

Lymphangioleiomyomatosis LAM is a female predominant and devastating pulmonary disease, characterized by diffusely infiltrated smooth muscle like cells that carry mutations in the tuberous sclerosis complex (TSC) genes. TSC1, TSC2 and TBC1D7 interact and inhibit the mammalian target of rapamycin complex 1 (mTORC1). The reasons that LAM exclusively affects women and how TSC1 or TSC2 deficiency contributes to the pathogenesis of LAM are not yet fully understood. We previously discovered that estrogen promotes the survival and lung metastases of tuberin-deficient rat uterine leiomyoma-derived ELT3 cells in a xenograft tumor model (PNAS 2009). Recently, we reported that estrogen and mTORC2 coordinate to enhance prostaglandin biosynthesis and tumorigenesis in LAM (J. Expt. Med. 2013). Prostaglandins are lipid mediators that participate in tumor survival, growth, invasion, and inflammation. Phospholipase A2 (PLA2), Cyclooxygenase-2 (COX-2) and prostacyclin synthase (PTGIS) are critical enzymes responsible for the production of prostaglandins. Prostaglandin receptors (EPs) mediate the biological function of prostaglandins. This study is to determine whether suppression of prostaglandin biosynthesis pathway potentially leads to tumor regression LAM in both cell culture and preclinical models of LAM. To identify additional pathways activated by TSC loss, we performed bioinformatics analysis of public expression arrays and found a rapamycin-insensitive upregulation of prostaglandin biosynthesis genes including (PLA2), cyclooxygenase-2 (COX-2), prostacyclin synthase (PTGIS), and prostaglandin E2 (PGE2) receptor 3 (EP3), in TSC2-deficient LAM patient-derived cells compared to TSC2-addback cells. Real-time RT-PCR assays validated the enhanced expression of PLA2, COX-2, PTGIS and EP3 in TSC2-deficient cells. Immunoblotting analysis showed the increased levels of PLA2, COX-2, PTGIS and EP3 in TSC2-deficient cells compared to TSC2-addback cells. Immunohistochemistry demonstrated abundant accumulation of PLA2, COX-2 and EP3 in LAM lung lesions compared to adjacent normal tissues. Interestingly, PGE2 specifically stimulated the growth of TSC2-deficient LAM patient-derived cells compared to TSC2-addback cells. Importantly, treatment of TSC2-deficient LAM patient-derived cells with PLA2 inhibitor or EP3 inhibitor more potently reduced cell proliferation in dose-dependent manner compared to TSC2-addback cells. Our data documents that loss of TSC2 leads to the aberrant expression and accumulation of prostaglandin biosynthesis regulators PLA2, COX-2, PTGIS and EP3, thereby enhancing prostaglandin production and promoting TSC-related cell growth and tumor development. Our data supports the potential application of prostaglandin metabolites as biomarkers of disease severity and the development of prostaglandin biosynthesis inhibitors as alternative therapeutic options for lesions occurring in LAM patients and in other gender-specific neoplasm. Citation Format: Chenggang Li, Po-Shun Lee, Yang Sun, Erik Zhang, Xiaoxiao Gu, Jing Li, Kai-Feng Xu, Alfredo Csibi, John Blenis, Elizabeth Petri Henske, Bruce Levy, David Kwiatkowski, Jane J. Yu. Estradiol and mTORC2 cooperate to enhance prostaglandin biosynthesis and tumorigenesis in tuberous sclerosis complex. [abstract]. In: Proceedings of the AACR Special Conference: Targeting the PI3K-mTOR Network in Cancer; Sep 14-17, 2014; Philadelphia, PA. Philadelphia (PA): AACR; Mol Cancer Ther 2015;14(7 Suppl):Abstract nr B10.

Regulation of organ size is achieved through the action of the mTOR and Hippo signalling pathways, which control cell proliferation and cell growth in response to extracellular cues. A link between these pathways is revealed by the finding that YAP downregulates PTEN to promote cell growth and tissue hyperplasia.

Corrigendum to “Endothelial relaxation mechanisms and nitrative stress are partly restored by vitamin D3 therapy in a rat model of polycystic ovary syndrome” [Life Sci. 93 (2013) 133–138] Gabriella Masszi, Rita Benko⁎, Noemi Csibi, Eszter M. Horvath, Anna-Maria Tokes, Agnes Novak, Nora Judit Beres, Robert Tarszabo, Anna Buday, Csaba Repas, Gabor Bekesi, Attila Patocs, Gyorgy L. Nadasy, Peter Hamar, Zoltan Benyo, Szabolcs Varbiro

Lymphangioleiomyomatosis (LAM) is a female predominant progressive and neoplastic disorder that leads to lung destruction and respiratory failure primarily in women. LAM is typically due to TSC2 mutations resulting in mTORC1 activation in proliferative smooth muscle-like cells in the lung. The female predominance of LAM suggests that estradiol contributes to disease development. Metabolomic profiling identified estradiol-enhanced prostaglandin biosynthesis signature in Tsc2-deficient cells, both in vitro and in vivo. Estradiol increased the expression of cyclooxygenase-2 (COX-2), a rate-limiting enzyme in prostaglandin biosynthesis, which was increased at baseline in TSC2-deficient cells, and was not affected by rapamycin treatment, but decreased by Torin 1 treatment and Rictor knockdown, associated with reduction of levels phospho-Akt (S473). Prostaglandin production was also increased in TSC2-deficient cells. In preclinical models, Celecoxib or aspirin treatment suppressed tumor progression. LAM patients had significantly higher serum prostaglandin levels than healthy women. 15-epi-lipoxin-A4 was identified in exhaled breath condensate from LAM subjects and was increased by aspirin treatment, indicative of functional COX-2 expression in the LAM airway. Targeting COX-2 and prostaglandin pathways may have therapeutic value in LAM and TSC-related diseases, and possibly in other conditions associated with mTOR-hyperactivation. Citation Format: Chenggang Li, Po-Shun Lee, Xiaoxiao Gu, Yang Sun, Erik Zhang, Alfredo Csibi, Jing Li, Carmen Priolo, Yanan Guo, Chin-Lee Wu, John Blenis, Kai-Feng Xu, Elizabeth Petri Henske, Bruce Levy, David Kwiatkowski, Jane J. Yu. Estradiol and mTORC2 orchestrate to enhance prostaglandin biosynthesis andtumorigenesis in tuberous sclerosis complex. [abstract]. In: Proceedings of the Third AACR International Conference on Frontiers in Basic Cancer Research; Sep 18-22, 2013; National Harbor, MD. Philadelphia (PA): AACR; Cancer Res 2013;73(19 Suppl):Abstract nr C59.

The mTOR complex 1 (mTORC1) is a ser/thr protein kinase complex and critical regulator of cellular metabolism, protein synthesis, autophagy and cell growth. This signaling system is normally carefully controlled by the integration of inputs from growth factors, nutrients, cellular energy status, oxygen availability and stress. A variety of loss-of-function or gain-of-function alterations in key mTORC1 regulatory proteins, very often results in inappropriate regulation of this signaling system. Uncontrolled mTORC1 activation contributes on its own or in combination with other pathways to metabolic diseases ranging from diabetes and obesity, cardiac hypertrophy, neurodegeneration, aging, benign tumor syndromes to highly malignant cancers. Mechanistically, uncontrolled mTORC1 activation leads to a constant positive input into anabolic processes and suppression of many catabolic processes. As a result, these cells require increased energy and carbon sources to meet their high metabolic needs for cell growth. To meet this demand, we have found in cells that are PTEN or TSC1/2 null, that activated mTORC1 uses distinct mechanisms to increase glutamine consumption (glutaminolysis) by elevating the expression of glutaminase and the activity of glutamate dehydrogenase. Furthermore, under nutrient-rich conditions, these tumor cells sense the increased energy production and in a positive feedback loop, promote more mTORC1 assembly via an AMP kinase (AMPK)-dependent, and a novel AMPK-independent mechanism. This acquired addiction to glutamine provides a novel therapeutic strategy for treating patients with activated mTORC1. Indeed, by acutely blocking the ability of energetically stressed cells with hyperactive mTORC1 to metabolize glutamine, we can selectively kill some tumor cells without damaging normal cells. We anticipate that these studies will lead to the development of “synthetic-lethal” drugs useful in the treatment of patients with diseases linked to inappropriate activation of mTORC1. Research supported by NIH RO1-GM51405 and the LAM Foundation. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr SY29-01. doi:1538-7445.AM2012-SY29-01

Anne Poupon: Biologie et Bioinformatique des Systèmes de Signalisation, UMR Physiologie de la Reproduction et des Comportements, INRA, Nouzilly, France. Sanchez A.M.J. : Equipe Remodelage Musculaire et Signalisation Contact: leibovs@supagro.inra.fr; The mTORC1 pathway is required for both the terminal muscle differentiation and hypertrophy by controlling the mammalian translational machinery via phosphorylation of S6K1 and 4E-BP1. mTOR and S6K1 are connected by interacting with the eIF3 initiation complex. The regulatory subunit eIF3f plays a major role in muscle hypertrophy and is a key target that accounts for MAFbx function during atrophy. Here we present evidence that in MAFbx-induced atrophy the degradation of eIF3f suppresses S6K1 activation by mTOR, whereas an eIF3f mutant insensitive to MAFbx polyubiquitination maintained persistent phosphorylation of S6K1 and rpS6. During terminal muscle differentiation a conserved TOS motif in eIF3f connects mTOR/raptor complex, which phosphorylates S6K1 and regulates downstream effectors of mTOR and Cap-dependent translation initiation. Thus eIF3f plays a major role for proper activity of mTORC1 to regulate skeletal muscle size.

International audience; Myostatin deficiency leads in skeletal muscle overgrowth but the precise molecular mechanisms underlying this hypertrophy are not well understood. In this study, to gain insight into the role of endogenous myostatin in the translational regulation, we used an in vitro model of cultured satellite cells derived from myostatin knock-out mice. Our results show that myostatin knock-out myotubes are larger than control myotubes and that this phenotype is associated with an increased activation of the Akt/mTOR signaling pathway, a known regulator of muscle hypertrophy. These results demonstrate that hypertrophy due to myostatin deficiency is preserved in vitro and suggest that myostatin deletion results in an increased protein synthesis. Accordingly, the rates of global RNA content, polysome formation and protein synthesis are all increased in myostatin-deficient myotubes while they are counteracted by the addition of recombinant myostatin. We furthermore demonstrated that genetic deletion of myostatin stimulates cap-dependent translation by positively regulating assembly of the translation preinitiation complex. Together the data indicate that myostatin controls muscle hypertrophy in part by regulating protein synthesis initiation rates, that is, translational efficiency.