Regression of the inflammatory microenvironment of the peritoneal cavity in women with endometriosis by GnRHa treatment.

OBJECTIVE To investigate the effect of gonadotropin-releasing hormone analogues (GnRHa) on the peritoneal fluid microenvironment in women with endometriosis. STUDY DESIGN Peritoneal fluid was collected from 85 women with severe endometriosis (rAFS stage III and IV) during laparoscopic surgery during the proliferative phase. Prior to surgery clinical data were collected. The concentrations of specific markers for endometriosis in the peritoneal fluid were determined using an ELISA and a comparison between peritoneal fluid markers in women using GnRHa and no hormonal treatment was performed using a non-parametric Mann-Whitney U test. RESULTS The study included peritoneal fluid from 39 patients who had been administered GnRHa (Zoladex(®)) in the three months prior to surgery and 46 from women with no hormonal treatment in this period. Concentrations of IL-8, PAPP-A, glycodelin-A and midkine were significantly reduced in the GnRHa treatment group compared to women receiving no hormonal treatment. RANTES, MCP-1, ENA-78, TNF-α, OPG, IP-10 and defensin showed no significant change between the two groups. CONCLUSIONS GnRHa mediate a significant regression in the inflammatory nature of the peritoneal microenvironment in women with endometriosis.

Materno-fetal nutrient transfer across primary human trophoblast layer.

MATERNO-FETAL NUTRIENT TRANSFER ACROSS PRIMARY HUMAN TROPHOBLAST MONOLAYER Objectives: Polarized trophoblasts represent the transport and metabolic barrier between the maternal and fetal circulation. Currently human placental nutrient transfer in vitro is mainly investigated unidirectionallyon cultured primary trophoblasts, or bidirectionally on the Transwell® system using BeWo cells treated with forskolin. As forskolin can induce various gene alterations (e.g. cAMP response element genes), we aimed to establish a physiological primary trophoblast model for materno-fetal nutrient exchange studies without forskolin application. Methods: Human term cytotrophoblasts were isolated by enzymatic digestion and Percoll® gradient separation. The purity of the primary cells was assessed by flow cytometry using the trophoblast-specific marker cytokeratin-7. After screening different coating matrices, we optimized the growth conditions for the primary cytotrophoblasts on Transwell/ inserts. The morphology of 5 days cultured trophoblasts was determined by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Membrane makers were visualized using confocal microscopy. Additionally transport studies were performed on the polarized trophoblasts in the Transwell® system. Results: During 5 days culture, the trophoblasts (>90% purity) developed a modest trans-epithelial electrical resistance (TEER) and a sizedependent apparent permeability coefficient (Papp) to fluorescently labeled compounds (MW ~400-70’000D). SEM analyses confirmed a confluent trophoblast layer with numerous microvilli at day six, and TEM revealed a monolayer with tight junctions. Immunocytochemistry on the confluent trophoblasts showed positivity for the cell-cell adhesion molecule E-cadherin, the tight junction protein ZO-1, and the membrane proteins ABCA1 and Na+/K+-ATPase. Vectorial glucose and cholesterol transport studies confirmed functionality of the cultured trophoblast barrier. Conclusion: Evidence from cell morphology, biophysical parameters and cell marker expressions indicate the successful and reproducible establishment of a primary trophoblast monolayer model suitable for transport studies. Application of this model to pathological trophoblasts will help to better understand the mechanism underlying gestational diseases, and to define the consequences of placental pathology on materno-fetal nutrient transport.

A computer-assisted microscopic analysis of bone tissue developed inside a polyactive plymer implanted into an equine articular surface

One of the most promising applications for the restoration of small or moderately sized focal articular lesions is mosaicplasty (MP). Although recurrent hemarthrosis is a rare complication after MP, recently, various strategies have been designed to find an effective filling material to prevent postoperative bleeding from the donor site. The porous biodegradable polymer Polyactive (PA; a polyethylene glycol terephthalate - polybutylene terephthalate copolymer) represents a promising solution in this respect. A histological evaluation of the longterm PA-filled donor sites obtained from 10 experimental horses was performed. In this study, attention was primarily focused on the bone tissue developed in the plug. A computer-assisted image analysis and quantitative polarized light microscopic measurements of decalcified, longitudinally sectioned, dimethylmethylene blue (DMMB)- and picrosirius red (PS) stained sections revealed that the coverage area of the bone trabecules in the PA-filled donor tunnels was substantially (25%) enlarged compared to the neighboring cancellous bone. For this quantification, identical ROIs (regions of interest) were used and compared. The birefringence retardation values were also measured with a polarized light microscope using monochromatic light. Identical retardation values could be recorded from the bone trabeculae developed in the PA and in the neighboring bone, which indicates that the collagen orientation pattern does not differ significantly among these bone trabecules. Based on our new data, we speculate that PA promotes bone formation, and some of the currently identified degradation products of PA may enhance osteo-conduction and osteoinduction inside the donor canal.

Evaluation of a Novel Screw Position in a Type III Distal Phalanx Fracture Model: An Ex Vivo Study

OBJECTIVE: Mechanical evaluation of a novel screw position used for repair in a type III distal phalanx fracture model and assessment of solar canal penetration (SCP). STUDY DESIGN: Experimental study. SAMPLE POPULATION: Disarticulated equine hooves (n = 24) and 24 isolated distal phalanges. METHODS: Hooves/distal phalanges cut in a sagittal plane were repaired with 1 of 2 different cortical screw placements in lag fashion. In group 1 (conventional screw placement), the screw was inserted halfway between the proximal border of the solar canal (SC) and the subchondral bone surface on a line parallel to the dorsal cortex, whereas in group 2, the screw was inserted more palmar/plantar, where a perpendicular line drawn from the group 1 position reached the palmar/plantar cortex. Construct strength was evaluated by 3-point bending to failure. SCP was assessed by CT imaging and macroscopically. RESULTS: Screws were significantly longer in group 2 and in forelimbs. Group 2 isolated distal phalanges had a significantly more rigid fixation compared with the conventional screw position (maximum point at failure 31%, bending stiffness 41% higher). Lumen reduction of the SC was observed in 13/52 specimens (all from group 2), of which 9 were forelimbs. CONCLUSIONS: More distal screw positioning compared with the conventionally recommended screw position for internal fixation of type III distal phalangeal fractures allows placement of a longer screw and renders a more rigid fracture fixation. The novel screw position, however, carries a higher risk of SCP

A RELIGIÃO NO CONFLITO ENTRE ISRAEL E PALESTINA NO CONTEXTO DA CRIAÇÃO DO ESTADO JUDAICO: ASPECTOS HISTÓRICOS (1896-1948)

O presente trabalho analisa o papel da religião no conflito entre Israel e Palestina, principalmente no contexto da implantação do Estado de Israel, em 1948. A análise toma como delimitação histórica do conflito o período de 1896 a 1948, quando ocorre a migração das primeiras levas de judeus para os territórios palestinos. A pergunta inicial é sobre como judeus e muçulmanos se relacionavam nos primeiros anos de imigração até a criação do Estado de Israel. O problema principal a ser esclarecido é como a construção cultural ocidental em relação aos palestinos interferiu no conflito, principalmente no que tange à tomada da terra e à construção de um novo país dentro de um já existente, socialmente, religiosamente e culturalmente. Finalmente a pesquisa pergunta pela repercussão do conflito entre israelenses e palestinos no campo religioso protestante, principalmente entre grupos conservadores e fundamentalistas deste ramo do cristianismo. A pesquisa é totalmente bibliográfica e toma como referência as teorias pós-coloniais para debater a história do território, no que se refere aos aspectos religiosos do conflito.

Definition of amide protection factors for early kinetic intermediates in protein folding

Hydrogen–deuterium exchange experiments have been used previously to investigate the structures of well defined states of a given protein. These include the native state, the unfolded state, and any intermediates that can be stably populated at equilibrium. More recently, the hydrogen–deuterium exchange technique has been applied in kinetic labeling experiments to probe the structures of transiently formed intermediates on the kinetic folding pathway of a given protein. From these equilibrium and nonequilibrium studies, protection factors are usually obtained. These protection factors are defined as the ratio of the rate of exchange of a given backbone amide when it is in a fully solvent-exposed state (usually obtained from model peptides) to the rate of exchange of that amide in some state of the protein or in some intermediate on the folding pathway of the protein. This definition is straightforward for the case of equilibrium studies; however, it is less clear-cut for the case of transient kinetic intermediates. To clarify the concept for the case of burst-phase intermediates, we have introduced and mathematically defined two different types of protection factors: one is Pstruc, which is more related to the structure of the intermediate, and the other is Papp, which is more related to the stability of the intermediate. Kinetic hydrogen–deuterium exchange data from disulfide-intact ribonuclease A and from cytochrome c are discussed to explain the use and implications of these two definitions.

Establishment of a confluent monolayer model with human primary trophoblast cells: novel insights into placental glucose transport

STUDY HYPOTHESIS Using optimized conditions, primary trophoblast cells isolated from human term placenta can develop a confluent monolayer in vitro, which morphologically and functionally resembles the microvilli structure found in vivo. STUDY FINDING We report the successful establishment of a confluent human primary trophoblast monolayer using pre-coated polycarbonate inserts, where the integrity and functionality was validated by cell morphology, biophysical features, cellular marker expression and secretion, and asymmetric glucose transport. WHAT IS KNOWN ALREADY Human trophoblast cells form the initial barrier between maternal and fetal blood to regulate materno-fetal exchange processes. Although the method for isolating pure human cytotrophoblast cells was developed almost 30 years ago, a functional in vitro model with primary trophoblasts forming a confluent monolayer is still lacking. STUDY DESIGN, SAMPLES/MATERIALS, METHODS Human term cytotrophoblasts were isolated by enzymatic digestion and density gradient separation. The purity of the primary cells was evaluated by flow cytometry using the trophoblast-specific marker cytokeratin 7, and vimentin as an indicator for potentially contaminating cells. We screened different coating matrices for high cell viability to optimize the growth conditions for primary trophoblasts on polycarbonate inserts. During culture, cell confluency and polarity were monitored daily by determining transepithelial electrical resistance (TEER) and permeability properties of florescent dyes. The time course of syncytia-related gene expression and hCG secretion during syncytialization were assessed by quantitative RT-PCR and enzyme-linked immunosorbent assay, respectively. The morphology of cultured trophoblasts after 5 days was determined by light microscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Membrane makers were visualized using confocal microscopy. Additionally, glucose transport studies were performed on the polarized trophoblasts in the same system. MAIN RESULTS AND THE ROLE OF CHANCE During 5-day culture, the highly pure trophoblasts were cultured on inserts coated with reconstituted basement membrane matrix . They exhibited a confluent polarized monolayer, with a modest TEER and a size-dependent apparent permeability coefficient (Papp) to fluorescently labeled compounds (MW ∼400-70 000 Da). The syncytialization progress was characterized by gradually increasing mRNA levels of fusogen genes and elevating hCG secretion. SEM analyses confirmed a confluent trophoblast layer with numerous microvilli, and TEM revealed a monolayer with tight junctions. Immunocytochemistry on the confluent trophoblasts showed positivity for the cell-cell adhesion molecule E-cadherin, the tight junction protein 1 (ZO-1) and the membrane proteins ATP-binding cassette transporter A1 (ABCA1) and glucose transporter 1 (GLUT1). Applying this model to study the bidirectional transport of a non-metabolizable glucose derivative indicated a carrier-mediated placental glucose transport mechanism with asymmetric kinetics. LIMITATIONS, REASONS FOR CAUTION The current study is only focused on primary trophoblast cells isolated from healthy placentas delivered at term. It remains to be evaluated whether this system can be extended to pathological trophoblasts isolated from diverse gestational diseases. WIDER IMPLICATIONS OF THE FINDINGS These findings confirmed the physiological properties of the newly developed human trophoblast barrier, which can be applied to study the exchange of endobiotics and xenobiotics between the maternal and fetal compartment, as well as intracellular metabolism, paracellular contributions and regulatory mechanisms influencing the vectorial transport of molecules. LARGE-SCALE DATA Not applicable. STUDY FUNDING AND COMPETING INTERESTS This study was supported by the Swiss National Center of Competence in Research, NCCR TransCure, University of Bern, Switzerland, and the Swiss National Science Foundation (grant no. 310030_149958, C.A.). All authors declare that their participation in the study did not involve factual or potential conflicts of interests.

1987 University of Michigan Football Team

1995 University of Michigan Baseball Team

Back Row: Student Manager Tim Freehan, Groundskeeper Erich Keil, Volunteer Coach Alan Trammell, Dan Sanborn, Brad Schemer, Alex Wozniak, Matt Herr, Chris Hesse, Bryan Besco, Derek Besco, John Papp, Mike Hribernik, Mike Cervenak, Mario Garza Jr., Dan Murphy, Trainer Rex Thompson, Student Trainer Stephanie Rummel. Missing: Chuck Winters.

Middle Row: Assistant Coach Ace Adams, Kirk Beermann, Mike Haskell, Matt Fleury, John Arvai, Kelly Dransfeldt, Tyler Steketee, Brian Steinbach, Andy Wade, Mark Temple, Mike Muir, Brad Tinkham, Mick Kalahar, Assistant Coach Steve Merriman.

Front Row: (From Left): Student Manager Matt Hyde, Brian Simmons, Scott Niemiec, Sean Coston, Jasen Livingston, Co-Captain Ryan Van Oeveren, Head Coach Bill Freehan, Co-Captain Rodney Goble, Aaron Toth, Matt Ferullo, Chad Chapman, Matt Humbles, Scott Weaver.

1996 University of Michigan Baseball Team

Back Row: (17) Manager Andy Galbreath, Manager Jeff Olsen, Groundskeeper Erich Keil, Andy Hood, Quinn DeMarrais, Alex Wozniak, Mike Seestedt, Brian Bush, Brian Kalczynski, Ryan Kelley, Pete Martay, Luke Bonner, Bobby Scales, Jason Alcaraz, Mario Garza Jr., Dan Murphy, Student Trainer Jon Nichols.

Middle Row: Assistant SID Jim Schneider, Mike Cervenak, John Papp, Bryan Besco, Derek Besco, Chuck Taylor, Tyler Steketee, J.J. Putz, Brian Steinbach, Mike Hribernik, Brian Berryman, Chris Hesse, Matt Herr, Brad Scheiner, Mick Kalahar, Trainer Rex Thompson.

Front Row: (13) Kirk Beermann, Marlon Wright, Mike Muir, John Arvai, Assistant Coach Tom Dodge, Assistant Coach Chris Harrison, Head Coach Geoff Zahn, Assistant Coach Ace Adams, Matt Hyde, Mark Temple, Matt Fleury, Kelly Dransfeldt, Mike Haskell.

1997 University of Michigan Baseball Team

Back Row: (16) Groundskeeper Erich Keil, Student Manager Jeff Singer, Kevin Quinn, Dan Murphy, Dan Sanborn, Jeff Van Sickle, Andrew Miller, Brad Scheiner, Rob Bobeda, Bryce Ralston, Alex Wozniak, Stephen Lenick, Brian Coughlin, Andy Hood, Trainer Rex Thompson, Student Trainer Karen Dunn.

Middle Row: (14) Jason Alcaraz, Luke Bonner, Pete Martay, Brian Bush, Matt Herr, Mike Hribernik, J.J. Putz, Bryan Cranson, Brian Berryman, Ryan Kelley, John Papp, Mike Seestedt, Bobby Scales, Mario Garza, Jr.

Front Row: (13) Tyler Steketee, Mike Cervenak, Brian Kalczynski, Marlon Wright, Brian Steinbach, Assistant Coach Chris Harrison, Head Coach Geoff Zahn, Assistant Coach Matt Hyde, Captain Kirk Beerman, Mike Haskell, Mick Kalahar, Derek Besco, Bryan Besco.

1998 University of Michigan Baseball Team

4th Row: (12) Nick Alexander, Jeff Van Sickle, Mike Seestedt, David Parrish, Rob Bobeda, J.J. Putz, Bryan Cranson, Joe Young, Brian Berryman, John Papp, Ryan Kelley, Vince Pistilli.

3rd Row: (12) Trainer Rex Thompson, Jason Alcaraz, Pete Martay, Brian Bush, Robbie Reid, Matt Herr, Luke Bonner, Mike Haskell, Bryce Ralston, Bobby Scales, Dan Sanborn, Student Trainer Rich Wright.

2nd Row (12) Derek Besco, Mario Garza, Jr., Mike Hribernik, Tyler Steketee, Brian Steinbach , Assistant Coach Chris Harrison, Head Coach Geoff Zahn, Assistant Coach Matt Hyde, Brian Kalczynski, Mick Kalahar, Mike Cervenak, Bryan Besco.

Front Row: (11, Sitting) Student Manager Josh Taft, Seth Greene, Kevin Quinn, Bill LaRosa, Scott Tousa, C.J. Ghannam, Stephen Lenick, Andy Hood, Mike Norkus, Student Manager Jeff Singer, Groundskeeper Erich Keil.

1999 University of Michigan Baseball Team

4th Row: (12) Trainer Joel Pickerman, Nick Alexander, Vince Pistilli, Nate Wright, Kirk Taylor, Phil Lorbert, Jeff Trzos, Nick Bellows, Joe Young, Jeff Sandor, Mike Sokol, Student Trainer Jaye Peterson.

3rd Row: (12) Rob Bobeda, David Parrish, Bryan Cranson, Dan Sanborn, John Papp, Robbie Reid, Luke Bonner , Ryan Kelly, Pete Martay, Andy Hood, Bryce Ralston, Student Trainer Todd Sonquist.

2nd Row (11) Mike Seestedt, Brian Bush, Jason Alcaraz, Mike Cervenak, Assistant Coach Chris Harrison, Head Coach Geoff Zahn, Assistant Coach Matt Hyde, Assistant Coach John Edman, Bobby Scales, J.J. Putz, Bryan Besco.

Front Row: (10, Sitting) Student Manager Josh Taft, Bobby Korecky, Jay Dines, Scott Tousa, C.J. Ghannam, Kevin Quinn, Bill LaRosa, Dan Dombos, Aaron Wilkens, Student Manager Jeff Singer.