Expression and structural features of endoglin (CD105), a transforming growth factor beta1 and beta3 binding protein, in human melanoma.

Human endoglin (CD105) is a member of the transforming growth factor beta (TGF-beta) receptor family that binds TGF-beta1 and -beta3, but not TGF-beta2, on human endothelial cells. Immunohistochemical analyses demonstrated that CD105 is expressed on normal and neoplastic cells of the melanocytic lineage. The anti-CD105 MAb, MAEND3, stained 50, 25 and 34% of intradermal naevi, primary and metastatic melanomas investigated, respectively, and nine out of 12 melanoma cell lines. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed that CD105 expressed by melanoma cells consists of a homodimeric protein with an apparent molecular weight of 180 and 95 kDa under non-reducing and reducing conditions. Cross-linking of 125I-labelled TGF-beta1 to melanoma cells, Mel 97, by disuccinimidyl suberate (DSS) demonstrated that CD105 expressed on pigmented cells binds TGF-beta1; the pattern of binding of TGF-beta1 to melanoma cells was found to be similar to that of human umbilical vein endothelial cells. The addition of exogenous, bioactive TGF-beta1 significantly (P<0.05) inhibited the growth of CD105-positive melanoma cells, Mel 97, but did not affect that of CD105-negative melanoma cells, F0-1. These data, altogether, demonstrate that CD105 is expressed on pigmented cells and might play a functionally relevant role in the biology of human melanoma cells by regulating their sensitivity to TGF-betas

The transcriptional regulator ZNF398 mediates pluripotency and epithelial character downstream of TGF-beta in human PSCs

Human pluripotent stem cells (hPSCs) have the capacity to give rise to all differentiated cells of the adult. TGF-beta is used routinely for expansion of conventional hPSCs as flat epithelial colonies expressing the transcription factors POU5F1/OCT4, NANOG, SOX2. Here we report a global analysis of the transcriptional programme controlled by TGF-beta followed by an unbiased gain-of-function screening in multiple hPSC lines to identify factors mediating TGF-beta activity. We identify a quartet of transcriptional regulators promoting hPSC self-renewal including ZNF398, a human-specific mediator of pluripotency and epithelial character in hPSCs. Mechanistically, ZNF398 binds active promoters and enhancers together with SMAD3 and the histone acetyltransferase EP300, enabling transcription of TGF-beta targets. In the context of somatic cell reprogramming, inhibition of ZNF398 abolishes activation of pluripotency and epithelial genes and colony formation. Our findings have clear implications for the generation of bona fide hPSCs for regenerative medicine

Animal Performance with and without Supplements in Mombaça Guinea Grass Pastures during Dry Season.

The objectives were to: 1) evaluate the potential of the animal performance on Panicum maximum cv. Mombaça (mombaça guinea grass) pasture during dry season, and 2) determine the effects of protein or protein-energy supplementation on liveweight gain of steers grazing guinea grass pasture. A complete randomized block design was employed with three treatments and three replicates. Treatments included non-supplemented animals (NS) and animals supplemented with protein (PS) or protein-energy (PES), provided at 0.15% or 0.6% of body weight for 115 days, during dry period. We used 36 weaned calves from initial bodyweigh (LW) of 192 kg (±5 kg) on nine plots of mombaça guinea grass (1.25 ha each). Each month, animals were weighed and pastures sampled to estimate forage characteristics. The average daily gain (ADG) was greater (P = 0.0001) for cattle fed supplement than for cattle fed no supplement (250 g steer-1), and greater for protein-energy supplement (770 g steer-1) than for protein supplement (460 g steer-1). Mombaça guinea grass pastures with 45 cm height at the end of the wetseason have enough forage mass for maintenance throughout the dry season about 1.4 AU ha-1 (AU = 450 kg BW), and reasonable nutritive value (average of 8,1% of crude protein and 55,3% of in vitro organic matter digestibility) to provide small gains. Considering the nutritive value of Mombaça guinea grass during the dry period, protein and energy supplementation is required for weaned calves to optimize their performance

Anti-PSMA CAR-engineered NK-92 Cells: An Off-the-shelf Cell Therapy for Prostate Cancer

Prostate cancer (PCa) has become the most common cancer among males in Europe and the USA. Adoptive immunotherapy appears a promising strategy to control the advanced stages of the disease by specifically targeting the tumor, in particular through chimeric antigen receptor T (CAR-T) cell therapy. Despite the advancements of CAR-T technology in the treatment of hematological malignancies, solid tumors still represent a challenge. To overcome current limits, other cellular effectors than T lymphocytes are under study as possible candidates for CAR-engineered cancer immunotherapy. A novel approach involves the NK-92 cell line, which mediates strong cytotoxic responses against a variety of tumor cells but has no effect on non-malignant healthy counterparts. Here, we report a novel therapeutic approach against PCa based on engineering of NK-92 cells with a CAR recognizing the human prostate-specific membrane antigen (PSMA), which is overexpressed in prostatic neoplastic cells. More importantly, the potential utility of NK-92/CAR cells to treat PCa has not yet been explored. Upon CAR transduction, NK-92/CAR cells acquired high and specific lytic activity against PSMA-expressing prostate cancer cells in vitro, and also underwent degranulation and produced high levels of IFN-\u3b3 in response to antigen recognition. Lethal irradiation of the effectors, a safety measure requested for the clinical application of retargeted NK-92 cells, fully abrogated replication but did not impact on phenotype and short-term functionality. PSMA-specific recognition and antitumor activity were retained in vivo, as adoptive transfer of irradiated NK-92/CAR cells in prostate cancer-bearing mice restrained tumor growth and improved survival. Anti-PSMA CAR-modified NK-92 cells represent a universal, off-the-shelf, renewable, and cost-effective product endowed with relevant potentialities as a therapeutic approach for PCa immunotherapy

Desempenho animal em pasto de hìbrido interespecífico de Brachiaria ruzizizensis e Brachiaria brizantha.

Comissão organizadora: Grácia Maria Soares Rosinha, Alexandra Rocha de Oliveir, Rodrigo Carvalho Alva

Sting Millennium Suite: integrated software for extensive analyses of 3d structures of proteins and their complexes.

This article is intended to show how Sting Millennium Suite of programs can be useful in the study of protein structure and analysis of its function, emphasizing recent improvement introduced to SMS. The program has extensive built-in instructions and detailed easy-to-use help which user is invited to consult before and during SMS use.Na publicação: Paula R Kuser

Hyaluronan is a natural and effective immunological adjuvant for protein-based vaccines

One of the main goals of vaccine research is the development of adjuvants that can enhance immune responses and are both safe and biocompatible. We explored the application of the natural polymer hyaluronan (HA) as a promising immunological adjuvant for protein-based vaccines. Chemical conjugation of HA to antigens strongly increased their immunogenicity, reduced booster requirements, and allowed antigen dose sparing. HA-based bioconjugates stimulated robust and long-lasting humoral responses without the addition of other immunostimulatory compounds and proved highly efficient when compared to other adjuvants. Due to its intrinsic biocompatibility, HA allowed the exploitation of different injection routes and did not induce inflammation at the inoculation site. This polymer promoted rapid translocation of the antigen to draining lymph nodes, thus facilitating encounters with antigen-presenting cells. Overall, HA can be regarded as an effective and biocompatible adjuvant to be exploited for the design of a wide variety of vaccines

Efectos de diferentes suplementos en el rendimiento de novillos que pastorean guinea Mombasa (Megathyrsus maximus) durante el período seco.

ABSTRACT - To mitigate the low animal performance on Mombaça guineagrass pasture during the dry period, feeding 2 types of supplement to 2 genetic groups was evaluated. The experimental design was a randomized block design following a 2 × 2 factorial arrangement with 4 replications. The treatments consisted of feeding 2 levels of supplement (0.25 and 1.0% of body weight; BW), named low-cost supplement (LCS; US$11.75/steer) and high-cost supplement (HCS; US$ 62.80/steer), respectively, for 130 days (July–October; dry season) to 2 genetic groups: Caracu and F1 Senepol × Caracu. The steers were supplemented daily and weighed every 28 days. Pastures were evaluated monthly to estimate the herbage accumulation rate, herbage mass (HM), leaf, stem and dead material percentages and nutritive value. HM, morphological components and nutritive value were independent of supplement type fed (P>0.05). There were decreases in HM (3,720 to 3,205 kg DM/ha), daily herbage allowance (14.0 to 9.4 kg DM/100 kg BW) and leaf percentage (33.4 to 21.2%) and increase in dead material percentage (53.3 to 67.7%) throughout the experimental period. In vitro organic matter digestibility (59.9%), crude protein concentration (10.0%), neutral detergent fiber (72.1%) and acid detergent lignin (2.9%) remained constant from July to September but increased markedly in October. Steers supplemented with HCS performed better (P<0.05) than those which received LCS (1.005 vs. 0.565 kg liveweight gain/head/day, respectively). Regardless of supplement type, F1 Senepol × Caracu steers had greater average daily gains than pure Caracu steers (0.88 vs. 0.71 kg/hd/d, respectively). Feeding HCS to steers in the dry season would produce better performance than LCS and could reduce time to reach slaughter weight but weight changes during the subsequent wet season should be monitored to assess the extent of any compensatory gain by the low-cost group during this period to reduce the weight advantage of the high-cost group

Theoretical and experimental evidence for a post-perovskite phase of MgSiO3 in Earth's D" layer

The Earth's lower mantle is believed to be composed mainly of (Mg,Fe)SiO3 perovskite, with lesser amounts of (Mg,Fe)O and CaSiO3). But it has not been possible to explain many unusual properties of the lowermost 150 km of the mantle (the D" layer) with this mineralogy. Here, using ab initio simulations and high-pressure experiments, we show that at pressures and temperatures of the D" layer, MgSiO3 transforms from perovskite into a layered CaIrO3-type post-perovskite phase. The elastic properties of the post-perovskite phase and its stability field explain several observed puzzling properties of the D" layer: its seismic anisotropy, the strongly undulating shear-wave discontinuity at its top and possibly the anticorrelation between shear and bulk sound velocities.Comment: PUBLISHED IN Nature 430, 445-448 (2004