Abstract
The integrins and G protein-coupled receptors are both fundamental in cell biology. The cross talk between these two, however, is unclear. Here we show that β
3
integrins negatively regulate ...G protein-coupled signaling by directly inhibiting the Gα
13
-p115RhoGEF interaction. Furthermore, whereas β
3
deficiency or integrin antagonists inhibit integrin-dependent platelet aggregation and exocytosis (granule secretion), they enhance G protein-coupled RhoA activation and integrin-independent secretion. In contrast, a β
3
-derived Gα
13
-binding peptide or Gα
13
knockout inhibits G protein-coupled RhoA activation and both integrin-independent and dependent platelet secretion without affecting primary platelet aggregation. In a mouse model of myocardial ischemia/reperfusion injury in vivo, the β
3
-derived Gα
13
-binding peptide inhibits platelet secretion of granule constituents, which exacerbates inflammation and ischemia/reperfusion injury. These data establish crucial integrin-G protein crosstalk, providing a rationale for therapeutic approaches that inhibit exocytosis in platelets and possibly other cells without adverse effects associated with loss of cell adhesion.
Multidrug-resistant (MDR) Pseudomonas aeruginosa infections are a major clinical challenge. Many isolates are carbapenem resistant, which severely limits treatment options; thus, novel therapeutic ...combinations, such as imipenem-relebactam (IMI/REL), ceftazidime-avibactam (CAZ/AVI), ceftolozane-tazobactam (TOL/TAZO), and meropenem-vaborbactam (MEM/VAB) were developed. Here, we studied two extensively drug-resistant (XDR) P. aeruginosa isolates, collected in the United States and Mexico, that demonstrated resistance to IMI/REL. Whole-genome sequencing (WGS) showed that both isolates contained acquired GES β-lactamases, intrinsic PDC and OXA β-lactamases, and disruptions in the genes encoding the OprD porin, thereby inhibiting uptake of carbapenems. In one isolate (ST17), the entire C terminus of OprD deviated from the expected amino acid sequence after amino acid G388. In the other (ST309), the entire
gene was interrupted by an IS
1328 insertion element after amino acid D43, rendering this porin nonfunctional. The poor inhibition by REL of the GES β-lactamases (GES-2, -19, and -20; apparent
of 19 ± 2 μM, 23 ± 2 μM, and 21 ± 2 μM, respectively) within the isolates also contributed to the observed IMI/REL-resistant phenotype. Modeling of REL binding to the active site of GES-20 suggested that the acylated REL is positioned in an unstable conformation as a result of a constrained Ω-loop.
New Guinea is a biologically diverse island, with a unique geologic history and topography that has likely played a role in the evolution of species. Few island-wide studies, however, have examined ...the phylogeographic history of lowland species. The objective of this study was to examine patterns of phylogeographic variation of a common and widespread New Guinean bird species (Colluricincla megarhyncha). Specifically, we test the mechanisms hypothesized to cause geographic and genetic variation (e.g., vicariance, isolation by distance and founder-effect with dispersal). To accomplish this, we surveyed three regions of the mitochondrial genome and a nuclear intron and assessed differences among 23 of the 30 described subspecies from throughout their range. We found support for eight highly divergent lineages within C. megarhyncha. Genetic lineages were found within continuous lowland habitat or on smaller islands, but all individuals within clades were not necessarily structured by predicted biogeographic barriers. There was some evidence of isolation by distance and potential founder-effects. Mitochondrial DNA sequence divergence among lineages was at a level often observed among different species or even genera of birds (5-11%), suggesting lineages within regions have been isolated for long periods of time. When topographical barriers were associated with divergence patterns, the estimated divergence date for the clade coincided with the estimated time of barrier formation. We also found that dispersal distance and range size are positively correlated across lineages. Evidence from this research suggests that different phylogeographic mechanisms concurrently structure lineages of C. megarhyncha and are not mutually exclusive. These lineages are a result of evolutionary forces acting at different temporal and spatial scales concordant with New Guinea's geological history.
Chromatin “remodeling” is widely accepted as the mechanism that permits access to DNA by the transcription machinery. To date, however, there has been no experimental measurement of the changes in ...the kinetics and thermodynamics of the DNA–histone octamer association that are required to remodel chromatin so that transcription may occur. Here, we present the results of optical tweezer measurements that compare the kinetic and thermodynamic properties of nucleosomes composed of unmodified histones with those of nucleosomes that contain a mutant histone H4 (H4-R45H), which has been shown to allow SWI/SNF remodeling factor-independent transcription from the yeast HO promoter in vivo. Our measurements, carried out in a force-clamp mode, determine the force-dependent unwinding and rewinding rates of the nucleosome inner turn. At each force studied, nucleosomes containing H4-R45H unwind more rapidly and rewind more slowly than nucleosomes containing unmodified H4, indicating that the latter are the more stable. Extrapolation to forces at which the winding and unwinding rates are equal determines the absolute free energy of the nucleosome inner turn to be −32kBT for nucleosomes containing unmodified H4 and −27kBT for nucleosomes containing H4-R45H. Thus, the “loosening” or “remodeling” caused by this point mutation, which is demonstrated to be sufficient to allow transcriptional machinery access to the HO promoter (in the absence of other remodeling factors), is 5kBT. The correlation between the free energy of the nucleosome inner turn and the sin (SWI/SNF-independent) transcription suggests that, beyond partial unwinding, complete histone unwinding may play a role in transcriptional activation.
Display omitted
► Optical tweezers were used to measure the rates of nucleosome unwinding/rewinding. ► Sin mutant nucleosomes unwind faster and rewind slower than wild type. ► The absolute free energy of the nucleosome inner turn is calculated. ► This study provides benchmark for the role of nucleosomes in epigenetics.
Successful treatment of Acinetobacter baumannii infections require early and appropriate antimicrobial therapy. One of the first steps in this process is understanding which β-lactamase (bla) alleles ...are present and in what combinations. Thus, we performed WGS on 98 carbapenem-resistant A. baumannii (CR Ab). In most isolates, an acquired blaOXA carbapenemase was found in addition to the intrinsic blaOXA allele. The most commonly found allele was blaOXA-23 (n = 78/98). In some isolates, blaOXA-23 was found in addition to other carbapenemase alleles: blaOXA-82 (n = 12/78), blaOXA-72 (n = 2/78) and blaOXA-24/40 (n = 1/78). Surprisingly, 20% of isolates carried carbapenemases not routinely assayed for by rapid molecular diagnostic platforms, i.e., blaOXA-82 and blaOXA-172; all had ISAba1 elements. In 8 CR Ab, blaOXA-82 or blaOXA-172 was the only carbapenemase. Both blaOXA-24/40 and its variant blaOXA-72 were each found in 6/98 isolates. The most prevalent ADC variants were blaADC-30 (21%), blaADC-162 (21%), and blaADC-212 (26%). Complete combinations are reported.
The dynamic packaging of DNA into chromatin is a key determinant of eukaryotic gene regulation and epigenetic inheritance. Nucleosomes are the basic unit of chromatin, and therefore the accessible ...states of the nucleosome must be the starting point for mechanistic models regarding these essential processes. Although the existence of different unwound nucleosome states has been hypothesized, there have been few studies of these states. The consequences of multiple states are far reaching. These states will behave differently in all aspects, including their interactions with chromatin remodelers, histone variant exchange, and kinetic properties. Here, we demonstrate the existence of two distinct states of the unwound nucleosome, which are accessible at physiological forces and ionic strengths. Using optical tweezers, we measure the rates of unwinding and rewinding for these two states and show that the rewinding rates from each state are different. In addition, we show that the probability of unwinding into each state is dependent on the applied force and ionic strength. Our results demonstrate not only that multiple unwound states exist but that their accessibility can be differentially perturbed, suggesting possible roles for these states in gene regulation. For example, different histone variants or modifications may facilitate or suppress access to DNA by promoting unwinding into one state or the other. We anticipate that the two unwound states reported here will be the basis for future models of eukaryotic transcriptional control.
Island of opportunity Bower, Deborah S; Lips, Karen R; Amepou, Yolarnie ...
Frontiers in ecology and the environment,
August 2019, Letnik:
17, Številka:
6
Journal Article
Recenzirano
Odprti dostop
The amphibian chytrid fungus Batrachochytrium dendrobatidis (chytrid) has caused the most widespread, disease-induced declines and extinctions in vertebrates recorded to date. The largest ...climatically suitable landmass that may still be free of this fungus is New Guinea. The island is home to a sizeable proportion of the world’s known frog species (an estimated 6%), as well as many additional, yet-to-be-described species. Two decades of research on the chytrid fungus have provided a foundation for improved management of amphibian populations. We call for urgent, unified, international, multidisciplinary action to prepare for the arrival of B dendrobatidis in New Guinea, to prevent or slow its spread within the island after it arrives, and to limit its impact upon the island’s frog populations. The apparent absence of the fungus in New Guinea offers an opportunity to build capacity in advance for science, disease surveillance, and diagnosis that will have broad relevance both for non-human animal health and for public health.
While Australia's policy on foreign affairs and national security is always complex and many-faceted, it has come to be dominated by concerns about the rivalry between the political and economic ...super-powers. The US-China relationship is now at the centre of foreign policy and security considerations. How the relationship is understood has a major bearing on Australian governments' foreign affairs, defence, national security and trade relations decisions. The issue is particularly important at present because the position adopted by the Federal Labor government has crucial implications across all these policy areas.
Abstract Background context Recent studies have demonstrated cases of adjacent vertebral body osteolysis when assessing the effect of bone morphogenetic protein (BMP) on fusion rates. However, no ...study to date has evaluated the course of osteolysis at different periods. Purpose To determine the incidence and resolution of osteolysis associated with BMP used in transforaminal lumbar interbody fusions (TLIF). Study design Retrospective review. Patient sample All TLIF cases using BMP performed at one institution with routine postoperative computed tomography (CT) scans at defined intervals. Outcome measures Area of osteolysis and fusion as determined by CT scan. Methods We performed a retrospective analysis of all patients at our facility who underwent TLIF with BMP. Included were all patients who had obtained a CT scan within 48 hours of surgery, 3 to 6 months postoperatively, and 1 to 2 years postoperatively. Areas of osteolysis were defined as lucency within the vertebral body communicating with the interbody spacer that was not present on the immediately postoperative CT scan. Areas of osteolysis were measured in all three planes and the volume used for comparison of the 3 to 6 months CT scans with the greater than 1 year CT scan. Results Twenty-three patients who underwent TLIF with BMP had obtained CT scans at all time periods required for evaluation. Seventy-eight vertebral bodies/end plates were assessed for osteolysis (39 levels). The incidence of osteolysis 3 to 6 months postoperatively in the adjacent vertebral bodies was 54% compared with 41% at 1 to 2 years. The mean volume of osteolysis was at 0.216 cm3 at 1 to 2 years compared with 0.306 cm3 at 3 to 6 months (p=.082). The area/rate of osteolysis did not appear to significantly affect the rate of fusion or final outcome with an overall union rate of 83%. Conclusions The rate of osteolysis decreased at 1 year compared with 3 to 6 months, but only 24% of the vertebral bodies with evidence of osteolysis at 3 to 6 months completely resolved by 1 year.
PDF
