Depletion of mitochondria in mammalian cells through enforced mitophagy

Mitochondria are not only the 'powerhouse' of the cell; they are also involved in a multitude of processes that include calcium storage, the cell cycle and cell death. Traditional means of investigating mitochondrial importance in a given cellular process have centered upon depletion of mtDNA through chemical or genetic means. Although these methods severely disrupt the mitochondrial electron transport chain, mtDNA-depleted cells still maintain mitochondria and many mitochondrial functions. Here we describe a straightforward protocol to generate mammalian cell populations with low to nondetectable levels of mitochondria. Ectopic expression of the ubiquitin E3 ligase Parkin, combined with short-term mitochondrial uncoupler treatment, stimulates widespread mitophagy and effectively eliminates mitochondria. In this protocol, we explain how to generate Parkin-expressing, mitochondria-depleted cells from scratch in 23 d, as well as offer a variety of methods for confirming mitochondrial clearance. Furthermore, we describe culture conditions to maintain mitochondrial-depleted cells for up to 30 d with minimal loss of viability, for longitudinal studies. This method should prove useful for investigating the importance of mitochondria in a variety of biological processes

The King is Dead, Long Live the King: Entering A New Era of Stem Cell Research and Clinical Development

In mid November the biopharma industry was shocked by the announcement from Geron that they were ending work on embryonic stem cell research and therapy. For more than 10 years the public image of all stem cell research has been equated with embryonic stem cells. Unfortunately, a fundamentally important medical and financial fact was being ignored: embryonic stem cell therapy is extremely immature. In parallel to efforts in embryonic stem cell research and development, scientists and physicians in the field of adult stem cells realized that the natural role of adult stem cells in the body is to promote healing and to act like endogenous "repair cells" and, as a result, numerous companies have entered the field of adult stem cell therapy with the goal of expanding numbers of adult stem cells for administration to patients with various conditions. In contrast to embryonic stem cells, which are extremely expensive and potentially dangerous, adult cell cells are inexpensive and have an excellent safety record when used in humans. Many studies are now showing that adult stem cells are practical, patient-applicable, therapeutics that are very close to being available for incorporation into the practice of medicine. These events signal the entrance of the field of stem cells into a new era: an era where hype and misinformation no longer triumph over economic and medical realities

Tools for Quality Testing of Batches of Artifacts: The Cryogenic Thermometers for the LHC

In the processing of data series, such as in the case of the resistance R vs. temperature T calibrations of the thermometers (several thousands) necessary for the LHC new accelerator at CERN, it is necessary to use automatic methods for determining the quality of the acquired data and the degree of uniformity of the thermometer characteristics, that are of the semiconducting type. In addition, it must be determined if the calibration uncertainties comply with the specifications in the wide temperature range 1,6 - 300 K. Advantage has been taken of the fact that these thermometers represent a population with limited variability, to apply a Least Squares Method with Fixed Effect. This allows to fit the data of all the thermometers together, by taking into account the individuality of each thermometer in the model as a deviation from one of them taken as reference Ri = f(Ti) + bk0 + bk1 g(Tki) + bk1g(Tki)2 + ... where f(Ti) is the model valid for all i data and all k thermometers, while the subsequent part is the "fixed effect" model for the k-th thermometer, where g(T) is a suitable function of T. This method is shown in the paper applied to different stages of the data processing. First, for efficient compensation for the thermal drift occurring during acquisition, robust against the occurrence of outliers. Second, for detection of clusters of thermometers with inherently different characteristics. Finally, for optimisation of the calibration-point distribution

Phyto-additives in rainbow trout (Oncorhynchus mykiss) nutrition

Abstract. Fodder additives have been used for a long time with notable effects. Most of them are chemically synthesized and are antibiotics derivatives. The biggest problem following their use is the development of drug-resistant bacteria. In order to overcome this downside and to abide to the new food safety regulations, a search for natural alternatives has begun and the attention has been focused on plants or plants extracts, some used for millennia as natural remedies. As they are natural substances the negative impact on fish health and environment is negligible. Phyto-additives are a relatively new class of additives and the knowledge regarding their use or mode of action is sometimes partially or totally lacking. They have multiple effects; most notable being the antimicrobial, antioxidant, immunostimulating and bioproductive effects. Phyto-additives are a heterogeneous group of additives being extracted from bulbs, roots, leaves or fruits of different plants species; they are available in many forms: dry and/or crushed or as liquids (essential oils). Research has been carried out world-wide, focusing on cultured species (rainbow trout, tilapia, and catfish) using a wide variety of plants (garlic, onion, ginger, Echinacea, nettle, sage, cinnamon) demonstrating the positive effects of the phyto-additives used. Even if they exhibit a positive influence on many parameters, their use is dose dependent, as higher or lower doses could have inhibitory effect or no effects at all

Shrec'16 Track: Retrieval of Human Subjects from Depth Sensor Data

International audienceIn this paper we report the results of the SHREC 2016 contest on "Retrieval of human subjects from depth sensor data". The proposed task was created in order to verify the possibility of retrieving models of query human subjects from single shots of depth sensors, using shape information only. Depth acquisition of different subjects were realized under different illumination conditions, using different clothes and in three different poses. The resulting point clouds of the partial body shape acquisitions were segmented and coupled with the skeleton provided by the OpenNI software and provided to the participants together with derived triangulated meshes. No color information was provided. Retrieval scores of the different methods proposed were estimated on the submitted dissimilarity matrices and the influence of the different acquisition conditions on the algorithms were also analyzed. Results obtained by the participants and by the baseline methods demonstrated that the proposed task is, as expected, quite difficult, especially due the partiality of the shape information and the poor accuracy of the estimated skeleton, but give useful insights on potential strategies that can be applied in similar retrieval procedures and derived practical applications. Categories and Subject Descriptors (according to ACM CCS): I.4.8 [IMAGE PROCESSING AND COMPUTER VISION]: Scene Analysis—Shap

Central bisectionectomy – a feasible alternative for centrally located liver tumors

Clinica de Chirurgie Generală şi Transplant Hepatic, I.C.Fundeni, Bucureşti, România, Al XII-lea Congres al Asociației Chirurgilor „Nicolae Anestiadi” din Republica Moldova cu participare internațională 23-25 septembrie 2015Introducere: Bisectionectomia centrală reprezintă rezecția hepatică a segmentelor 4a, 4b, 5 și 8, definite astfel de sistemul Brisbane. Denumită anterior în diferite feluri (mesohepatectomie, hepatectomie centrală, lobectomie centrală) bisectionectomia centrală este operația de elecție pentru tumorile hepatice situate în segmentele centrale, și este de preferat rezecțiilor hepatice extinse (trisectionectomia dreaptă/stângă) pentru că minimalizează riscul insuficienței hepatice postoperatorii. Prezentare de caz: Pacienta în vârstă de 54 de ani cu dureri în hipocondrul drept, scădere ponderală cca. 8 kg / 2 luni. Bioumoral: AFP, CA 19.9 și ACE – în limite normale / AgHbs, Ac Anti-HBc, Ac Anti-HCV – negative. EDS/EDI – de aspect normal, prezintă la examenul CT o formațiune tumorală hepatică situată central (segmentele 4a, 4b, 5 și 8) ce înglobează VHM, la distanță de bifurcația venei porte. Se practică o bisectionectomie centrală, cu ligaturarea pediculilor vasculari glissonieni drept anterior (seg. 5 și 8) și stâng medial (seg. 4a și 4b). Transecțiunea parenchimului hepatic se efectuează folosindu-se “SONOPET”. Cu evoluție postoperatorie bună, pacienta se externează la 9 zile postoperator. La 6 luni postoperator probele serice AFP – în limite normale, fără semne imagistice de recidivă. Bisectionectomia centrală poate fi realizată atunci când tumora nu invadează pediculii vasculari aferenți parenchimului restant (secțiunea dreaptă posterioara / secțiunea stângă laterală) și presupune doua planuri de secțiune. Concluzii: Bisectionectomia centrală este operația de elecție pentru tumorile hepatice centrale prin care se prezervă cantitatea maxima de parenchim hepatic, minimalizându-se riscul de IHA postoperatorie. Este posibilă în cazuri atent selecționate și necesită experiență în rezecțiile hepatice majore.Introduction: By Brisbane terminology central bisectionectomy is the resection of segments 4a, 4b, 5, and 8 of the liver. Formerly called in different ways (mesohepatectomy, central hepatectomy, central lobectomy) central bisectionectomy is the elective surgery for liver tumors located in central segments (4a, 4b, 5 and 8), with better results than extensive liver resections (right / left trisectionectmy) because it minimizes the risk of postoperative liver failure. Case report: A 54 years old female, presented for right upper quadrant pain, weight loss (8 kg / 2 months); Blood tests: normal AFP; negative Ag Hbs, Ab Anti-HBc, Ab Anti-HCV. UGE/LGE – with a normal aspect; CT scan revealed centrally located liver tumor (segments 4a, 4b, 5, and 8) that includes MHV, away from the portal vein bifurcation. We performed a regulated central bisectionectomy with ligation of vascular pedicles for right anterior section (seg. 5 and 8) and those for left medial section (seg 4a and 4b). With good postoperative course, the patient was discharged in the 9th postoperative day. Six months postoperatively, the patient is good, without tumor relapse signs. Central bisectionectomy can not be performed when the tumor invades the vascular pedicles afferent to remaining parenchyma and requires two planes of transection. Conclusions: Central bisectionectomy is the best choice surgery for centrally located liver tumors which preserves the maximum amount of liver parenchyma thus minimizing the risk of postoperative liver failure

Fitting a 3D Morphable Model to Edges: A Comparison Between Hard and Soft Correspondences

We propose a fully automatic method for fitting a 3D morphable model to single face images in arbitrary pose and lighting. Our approach relies on geometric features (edges and landmarks) and, inspired by the iterated closest point algorithm, is based on computing hard correspondences between model vertices and edge pixels. We demonstrate that this is superior to previous work that uses soft correspondences to form an edge-derived cost surface that is minimised by nonlinear optimisation.Comment: To appear in ACCV 2016 Workshop on Facial Informatic

Living donor liver transplant in Romania – 116 cases experience

Centrul de Chirurgie Generală și Transplant Hepatic “Dan Setlacec”, Institutul Clinic Fundeni, Bucureşti, România, Al XII-lea Congres al Asociației Chirurgilor „Nicolae Anestiadi” din Republica Moldova cu participare internațională 23-25 septembrie 2015Introducere: Cererea pentru transplantul hepatic în Romania este în continuă creștere în timp ce penuria de donatori de organe persistă. Timpul lung de așteptare pentru un transplant de ficat şi disfuncţia hepatică progresivă aferentă așteptării a motivat multe familii să ia în considerare donarea de organe. Material și metode: În anul 2000 a fost realizat primul transplant hepatic ortotopic cu ficat întreg de către echipa I.Popescu et al. la Institutul Clinic Fundeni din Bucureşti, urmat de transplantul de ficat de la donator viu (LDLT) mai târziu în acelaşi an, ajungând în aprilie 2015 la un total de 116 de transplanturi cu ficat de la donator viu. LDLT a fost realizată cu hemificat drept la 78 pacienți, secțiune laterală stânga – la 26 pacienți, hemificat stâng – la 7 pacienți, hemificat stâng în bloc cu segmentul 1 – la 3 pacienți, şi dual graft – la 2 pacienți. Rezultate: Rata de morbidități majore a fost de 53,4% (62 pacienți), rata generală de retransplant a fost de 11,3% (13 pacienți). Rata de supraviețuire generală la 1-, 3-, şi 5 ani a fost de 88,8%, 82,5%, respectiv, 79,2%. Concluzii: Scopul nostru a fost de a reduce rata mortalității pe lista de așteptare prin scurtarea timpului de aşteptare pentru TH prin asigurarea grefelor necesare. Avantajele LDLT includ posibilitatea de a fi efectuate în regim programat şi cu un timp de aşteptare scurt pentru primitor, în timp ce indicaţiile de transplant pot fi extinse (ex: HCC în afara criteriilor Milano).Introduction: The request for Liver Transplant (LT) in Romania continues to increase while the donor pool size remains inadequate. The long waiting time for a liver transplant and the progressive liver dysfunction that occurs in this time has motivated many families to consider living donation. Material and methods: In 2000, the first successful LT (with whole graft) was carried out by I.Popescu et al. at Fundeni Clinical Institute in Bucharest, followed by the first living donor liver transplantation (LDLT) (successful) later the same year, reaching 116 living donors liver transplants by April 2015. LDLT was performed with right hemiliver in 78 pts, left lateral section in 26 pts, left hemiliver in 7 pts, left hemiliver with segment 1 in 3 pts, and dual graft LDLT in 2 pts: one received a right hemiliver and a left lateral section (17), and one received a left hemiliver and a left lateral section. Results: Major morbidity rates were 53.4% (62 pts) Overall retransplantation rate was 11.3% (13 pts). Long-term overall 1-, 3-, and 5- year estimated survival rates were 88.8%, 82.5%, and 79.2%, respective. Conclusions: Our main goal was to continuously reduce the drop-out rate on waiting list (due to mortality and morbidity) by shortening the waiting time for LT insured by providing the necessary grafts. Advantages of LDLT include the ability to be performed on an elective basis, with optimal timing and no waiting time for the recipient, while the indications for transplantation may be extended (i.e. HCC beyond Milan criteria)

15 years of Romanian national program of liver transplant – a retrospective analysis of 648 patients operated

Centrul de Chirurgie Generală şi Transplant Hepatic “Dan Setlacec”, Institutul Clinic Fundeni, Bucureşti, România, Al XII-lea Congres al Asociației Chirurgilor „Nicolae Anestiadi” din Republica Moldova cu participare internațională 23-25 septembrie 2015Introducere: Transplantul hepatic (TH) a devenit metoda de elecție în tratamentul bolilor hepatice în stadiu terminal, cu peste 20.000 de proceduri anuale la nivel mondial. Scopul acestui studiu este de a analiza rezultatele programului de transplant hepatic românesc după 15 ani de activitate. Material și metode: Între aprilie 2000 și aprilie 2015, 648 de pacienți au fost transplantați cu ficat în România. Raportul pe sexe a fost 382 (m) / 266 (f), în timp ce raportul adulți/copii a fost de 588/60, cu o vârstă medie de 45 de ani. Indicațiile principale pentru TH au fost: ciroza de etiologie VHB (176 pct; 27,1%), HCC (128 pct; 19,7%), și ciroza de etiologie VHC (118 pct; 18,2%). Transplantul hepatic de la donator cadaveric a fost efectuat la 532 pacienți (82,1%): 512 pct cu ficat întreg, 16 pct – cu ficat split, 2 pct – cu ficat redus, transplant-domino – la 2 pct. Transplantul de ficat de la donator viu a fost efectuat la 116 pct (17,9%). Rezultate: Mortalitatea perioperatorie a fost de 7,9% (51 pct). Rata de retransplant a fost de 4,6% (30 pct). Supraviețuirea la 1, 3 și 5 ani pe termen lung a fost estimată la 88,8%, 82,5% și respectiv 79,2%. Timpul de așteptare pentru TH a scăzut semnificativ în timp. Mortalitatea pe un an pe lista de așteptare a scăzut semnificativ, de la 31,4% la 11,8%. Concluzii: Programul de transplant hepatic în România se adresează atât cauzelor de insuficiență hepatică acută și cronică, cât și tumorilor hepatice la adulți și copii, fiind folosite toate tehnicile chirurgicale. Amploarea programului a crescut constant în timp, ceea ce a adus la scurtarea timpului și a ratelor de mortalitate pe lista de așteptare.Background: Liver transplantation (LT) is the treatment of choice for end-stage liver diseases, with more than 20.000 procedures yearly worldwide. The aim of this study was to analyze the results of Romanian National Program of LT after 15 years of activity. Methods: Between April 2000 and April 2015, 648 patients received 678 LTs in Romania. Male/female ratio was 382/266, while adult/pediatric ratio was 588/60, with a mean age range 7 months – 68 yr. Main LT indications in the adult population were HBVrelated cirrhosis (176 pts; 27.1%), hepatocellular carcinoma (128 pts; 19.7%), and HCV-related cirrhosis (118 pts; 18.2%). DDLT (death donor liver transplant) was performed in 532 pts (82.1%): whole LT in 512 pts, split LT in 16 pts, reduced LT in 2 pts, and domino LT in 2 pts. LDLT (living donor liver transplant) was performed in 116 pts (17.9%). Results: Perioperative mortality was 7.9% (51 pts). Retransplantation rate was 4.6% (30 pts). Long-term overall 1-, 3-, and 5- year estimated survival rates were 88.8%, 82.5%, and 79.2%, respectively. Median waiting time for LT decreased significantly over time. One-year overall mortality on waiting list also decreased significantly over time from 31.4% to 11.8%. Conclusions: The liver transplantation program in Romania addresses all causes of acute and chronic liver failure or liver tumors in adults and children, using all surgical techniques. The program constantly increased over time, leading to less time and lower mortality rate on the waiting list with the results similar to those of other centers