A rapid and simple method for constructing stable mutants of <Emphasis Type="Italic">Acinetobacter baumannii</Emphasis>

Background  

Acinetobacter baumannii is a multidrug-resistant bacterium responsible for nosocomial infections in hospitals worldwide. Study of mutant phenotypes is fundamental for understanding gene function. The methodologies developed to inactivate A. baumannii genes are complicated and time-consuming; sometimes result in unstable mutants, and do not enable construction of double (or more) gene knockout mutant strains of A. baumannii.     

Abundance patterns of the toxic phytoplankton in coastal waters of the Balearic Archipelago (NW Mediterranean Sea): a multivariate approach

The prevalence of catch-and-release factors known to adversely affect the mortality or physical condition of Australian bass Macquaria novemaculeata were surveyed across a range of anglers throughout impoundments and rivers in New South Wales. Subsamples of tournament-caught fish were also monitored in tanks for 1 h after being weighed to quantify immediate mortalities and sublethal physical damage. Most fish were caught on actively fished artificial baits, and were mouth hooked (96%), which resulted in no immediate mortality amongst monitored fish and a low frequency of mortality-causing factors in surveys. However, fish that were weighed-in during tournaments often had fin damage (52% of fish) and/or barotrauma (25% of fish, in impoundments only), and were held in live wells with poor water-quality. The prevalence of these sublethal effects varied considerably according to the specific seasons, locations, rules and procedures of each tournament; but could not be reliably attributed to any of the recorded catch-and-release variables (except for dissolved oxygen, which was significantly influenced by live-well volume). While these results validate the current release of angled Australian bass as a means of conserving their stocks, the potential for adverse effects could nevertheless be minimised via simple changes to conventional handling.     

蒙古中部湖泊之谷沉积岩-玄武岩共存的渐新世地层:蒙古-奥地利合作项目回顾(英文)

Taatsiin Gol和Taatsiin Tsagaan Nuur地区的渐新世沉积序列具有重要的地层学意义:这里出露的三达河组和Loh组沉积含有多个化石层和玄武岩夹层。在蒙古-奥地利合作项目中,从研究区域的33个剖面/化石地点的85个化石层中采集了289种化石(11种腹足类、2种两栖类、9种爬行类和267种哺乳类)。本文提供了所有地点的完整哺乳动物清单,并结合大、小哺乳动物的新资料,对蒙古非正式的生物带A,B,C和C1进行了更新。40Ar/39Ar测年给出了至少两组玄武岩年龄:早渐新世玄武岩I组大约31.5Ma,晚渐新世玄武岩II组大约28Ma。它们可以用作渐新世哺乳动物地层学的年代校正点。从早渐新世至晚渐新世,哺乳动物群发生了显著的变化,包括晚渐新世种数的明显减少。这种趋势在肉齿类、食肉类和反刍类中最为突出。     

Late Holocene palaeoecology of Lago Verde: evidence of human impact and climate change in the northern limit of the neotropics during the late formative and classic periods

Multiproxy analysis (pollen, diatom, charcoal) on a 6 m core from Lago Verde (Sierra de Los Tuxtlas), shows evidences of environmental changes and human impact on the evergreen rainforest on the tropical lowlands of eastern Mexico during the last ca. 2,800 years. Prehistoric human occupation is recorded since the late Formative throughout the middle Classic (250 b.c.–ca. a.d. 800) by the presence of maize pollen, a low abundance of tropical arboreal taxa and a high abundance of herbaceous pollen and charcoal particles. This occurred under a scenario of very low lake levels from which dry conditions are inferred based on the dominance of aerophilous and periphytic diatom taxa. After ca. a.d. 800 the site was abandoned and forest regeneration started, at the same time higher lake levels, an indication of more humid conditions, were established. In the absence of human disturbance, tropical forest regeneration was rapid (ca. 200 years). Fluctuations in pollen composition during times of low human population at the site are related to climate variability, with the highest tropical forest diversity and lake levels recorded during the Little Ice Age. Modern human impact is also recorded and compared with the prehistoric deforestation event. Comparison with palaeoecological records from Yucatan and the highlands of central Mexico offers a Mesoamerican perspective of climatic variability giving evidence that the late Formative and early to middle Classic demographic expansion occurred under a scenario of climates dryer than present, with the Postclassic characterized by moister conditions. The end of the Classic (ca. a.d. 800–1000) is identified as a period of rapid climate change which marks one of the most important cultural transitions in Mesoamerica.     

Potential use of the human amniotic membrane as a scaffold in human articular cartilage repair

The human amniotic membrane (HAM) is an abundant and readily obtained tissue that may be an important source of scaffold for transplanted chondrocytes in cartilage regeneration in vivo. To evaluate the potential use of cryopreserved HAMs as a support system for human chondrocytes in human articular cartilage repair. Chondrocytes were isolated from human articular cartilage, cultured and grown on the chorionic basement membrane side of HAMs. HAMs with chondrocytes were then used in 44 in vitro human osteoarthritis cartilage repair trials. Repair was evaluated at 4, 8 and 16 weeks by histological analysis. Chondrocytes cultured on the HAM revealed that cells grew on the chorionic basement membrane layer, but not on the epithelial side. Chondrocytes grown on the chorionic side of the HAM express type II collagen but not type I, indicating that after being in culture for 3–4 weeks they had not de-differentiated into fibroblasts. In vitro repair experiments showed formation on OA cartilage of new tissue expressing type II collagen. Integration of the new tissue with OA cartilage was excellent. The results indicate that cryopreserved HAMs can be used to support chondrocyte proliferation for transplantation therapy to repair OA cartilage.     

Overexpression of miR-128 specifically inhibits the truncated isoform of NTRK3 and upregulates BCL2 in SH-SY5Y neuroblastoma cells

Background

The CTCF insulator protein is a highly conserved zinc finger protein that has been implicated in many aspects of gene regulation and nuclear organization. The protein has been hypothesized to organize the human genome by forming DNA loops.

Results

In this paper, we report biochemical evidence to support the role for CTCF in forming DNA loops. We have measured DNA bending by CTCF at the chicken HS4 β-globin FII insulator element in vitro and have observed a unique DNA structure with aberrant electrophoretic mobility which we believe to be a DNA loop. CTCF is able to form this unusual DNA structure at two other binding sites: the c-myc P2 promoter and the chicken F1 lysozyme gene silencer. We also demonstrate that the length though not the sequence of the DNA downstream of the binding site is important for the ability of CTCF to form this unusual DNA structure. We hypothesize that a single CTCF protein molecule is able to act as a "looper" possibly through the use of several of its zinc fingers.

Conclusions

CTCF is able to form an unusual DNA structure through the zinc finger domain of the protein. This unusual DNA structure is formed in a directional manner by the CTCF protein. The findings described in this paper suggest mechanisms by which CTCF is able to form DNA loops, organize the mammalian genome and function as an insulator protein.     

FTO Genotype Is Associated With Exercise Training–induced Changes in Body Composition

The fat mass (FM) and obesity‐associated (FTO) gene is the first obesity‐susceptibility gene identified by genome‐wide association scans and confirmed in several follow‐up studies. Homozygotes for the risk allele (A/A) have 1.67 times greater risk of obesity than those who do not have the allele. However, it is not known whether regular exercise‐induced changes in body composition are influenced by the FTO genotype. The purpose of our study was to test whether the FTO genotype is associated with exercise‐induced changes in adiposity. Body composition was derived from underwater weighing before and after a 20‐week endurance training program in 481 previously sedentary white subjects of the HERITAGE Family Study. FTO single‐nucleotide polymorphism (SNP) rs8050136 was genotyped using Illumina GoldenGate assay. In the sedentary state, the A/A homozygotes were significantly heavier and fatter than the heterozygotes and the C/C homozygotes in men (P = 0.004) but not in women (P = 0.331; gene‐by‐sex interaction P = 0.0053). The FTO genotype was associated with body fat responses to regular exercise (P < 0.005; adjusted for age, sex, and baseline value of response trait): carriers of the C allele showed three times greater FM and %body fat losses than the A/A homozygotes. The FTO genotype explained 2% of the variance in adiposity changes. Our data suggest that the FTO obesity‐susceptibility genotype influences the body fat responses to regular exercise. Resistance to exercise‐induced reduction in total adiposity may represent one mechanism by which the FTO A allele promotes overweight and obesity.     

Antiangiogenic and Antitumor Effects of Trypanosoma cruzi Calreticulin

Background

In Latin America, 18 million people are infected with Trypanosoma cruzi, the agent of Chagas'' disease, with the greatest economic burden. Vertebrate calreticulins (CRT) are multifunctional, intra- and extracellular proteins. In the endoplasmic reticulum (ER) they bind calcium and act as chaperones. Since human CRT (HuCRT) is antiangiogenic and suppresses tumor growth, the presence of these functions in the parasite orthologue may have consequences in the host/parasite interaction. Previously, we have cloned and expressed T. cruzi calreticulin (TcCRT) and shown that TcCRT, translocated from the ER to the area of trypomastigote flagellum emergence, promotes infectivity, inactivates the complement system and inhibits angiogenesis in the chorioallantoid chicken egg membrane. Most likely, derived from these properties, TcCRT displays in vivo inhibitory effects against an experimental mammary tumor.

Methodology and Principal Findings

TcCRT (or its N-terminal vasostatin-like domain, N-TcCRT) a) Abrogates capillary growth in the ex vivo rat aortic ring assay, b) Inhibits capillary morphogenesis in a human umbilical vein endothelial cell (HUVEC) assay, c) Inhibits migration and proliferation of HUVECs and the human endothelial cell line Eahy926. In these assays TcCRT was more effective, in molar terms, than HuCRT: d) In confocal microscopy, live HUVECs and EAhy926 cells, are recognized by FITC-TcCRT, followed by its internalization and accumulation around the host cell nuclei, a phenomenon that is abrogated by Fucoidin, a specific scavenger receptor ligand and, e) Inhibits in vivo the growth of the murine mammary TA3 MTXR tumor cell line.

Conclusions/Significance

We describe herein antiangiogenic and antitumor properties of a parasite chaperone molecule, specifically TcCRT. Perhaps, by virtue of its capacity to inhibit angiogenesis (and the complement system), TcCRT is anti-inflammatory, thus impairing the antiparasite immune response. The TcCRT antiangiogenic effect could also explain, at least partially, the in vivo antitumor effects reported herein and the reports proposing antitumor properties for T. cruzi infection.