Red blood cell T-activation and hemolysis in surgical intensive care patients with severe infections

The exposure of Thomsen-Friedenreich (T) antigens on RBCs, serum neuraminidase, and serum hemoglobin levels were investigated in 53 adult surgical intensive care unit (ICU) patients with septicemia. Unmasked T-antigens were assayed by a hemagglutination test using peanut agglutinin (PNA) (direct anti-T test), and by an indirect anti-T test employing rabbit anti-PNA globulin. RBC T-activation was demonstrated in 17/53 patients (32%); in 2/53 patients (4%) the direct anti-T test was positive, indicating strong T-exposure. No polyagglutination phenomena were observed. Serum neuraminidase was elevated in 12/17 (71%) patients with T-activation and in 7/36 (19%) patients without T-activation. Free serum hemoglobin was elevated in 12/17 (71%) patients with T-activation and in 5/36 (14%) patients without T-activation. Correlations between T-activation and serum neuraminidase and between T-activation and serum hemoglobin were significant (p less than 0.001). Potentially neuraminidase-releasing bacteria were demonstrated in 13/17 (76%) patients with RBC T-exposure. We conclude that neuraminidase-induced RBC T-activation and subsequent hemolysis may be involved in the pathomechanism of hemolytic anemia in patients with severe infections.     

Configurational entropy change of netropsin and distamycin upon DNA minor-groove binding

Binding of a small molecule to a macromolecular target reduces its conformational freedom, resulting in a negative entropy change that opposes the binding. The goal of this study is to estimate the configurational entropy change of two minor-groove-binding ligands, netropsin and distamycin, upon binding to the DNA duplex d(CGCGAAAAACGCG).d(CGCGTTTTTCGCG). Configurational entropy upper bounds based on 10-ns molecular dynamics simulations of netropsin and distamycin in solution and in complex with DNA in solution were estimated using the covariance matrix of atom-positional fluctuations. The results suggest that netropsin and distamycin lose a significant amount of configurational entropy upon binding to the DNA minor groove. The estimated changes in configurational entropy for netropsin and distamycin are -127 J K(-1) mol(-1) and -104 J K(-1) mol(-1), respectively. Estimates of the configurational entropy contributions of parts of the ligands are presented, showing that the loss of configurational entropy is comparatively more pronounced for the flexible tails than for the relatively rigid central body.     

Characterization of extracellular vesicle miRNA identified in peripheral blood of chronic pancreatitis patients

Molecular and Cellular Biochemistry - Plasma-derived extracellular vesicles (EV) can serve as markers of cell damage/disease but can also have therapeutic utility depending on the nature of their...     

The contractile basis of amoeboid movement: V. The control of gelation, solation, and contraction in extracts from dictyostelium discoideum

Motile extracts have been prepared from Dictyostelium discoideum by homogenization and differential centrifugation at 4 degrees C in a stabilization solution (60). These extracts gelled on warming to 25 degrees Celsius and contracted in response to micromolar Ca++ or a pH in excess of 7.0. Optimal gelation occurred in a solution containing 2.5 mM ethylene glycol-bis (β-aminoethyl ether)N,N,N',N'-tetraacetate (EGTA), 2.5 mM piperazine-N-N'-bis [2-ethane sulfonic acid] (PIPES), 1 mM MgC1(2), 1 mM ATP, and 20 mM KCI at ph 7.0 (relaxation solution), while micromolar levels of Ca++ inhibited gelation. Conditions that solated the gel elicited contraction of extracts containing myosin. This was true regardless of whether chemical (micromolar Ca++, pH >7.0, cytochalasin B, elevated concentrations of KCI, MgC1(2), and sucrose) or physical (pressure, mechanical stress, and cold) means were used to induce solation. Myosin was definitely required for contraction. During Ca++-or pH-elicited contraction: (a) actin, myosin, and a 95,000-dalton polypeptide were concentrated in the contracted extract; (b) the gelation activity was recovered in the material sqeezed out the contracting extract;(c) electron microscopy demonstrated that the number of free, recognizable F-actin filaments increased; (d) the actomyosin MgATPase activity was stimulated by 4- to 10-fold. In the absense of myosin the Dictyostelium extract did not contract, while gelation proceeded normally. During solation of the gel in the absense of myosin: (a) electron microscopy demonstrated that the number of free, recognizable F- actin filaments increased; (b) solation-dependent contraction of the extract and the Ca++-stimulated MgATPase activity were reconstituted by adding puried Dictyostelium myosin. Actin purified from the Dictyostelium extract did not gel (at 2 mg/ml), while low concentrations of actin (0.7-2 mg/ml) that contained several contaminating components underwent rapid Ca++ regulated gelation. These results indicated : (a) gelation in Dictyostelium extracts involves a specific Ca++-sensitive interaction between actin and several other components; (b) myosin is an absolute requirement for contraction of the extract; (c) actin-myosin interactions capable of producing force for movement are prevented in the gel, while solation of the gel by either physical or chemical means results in the release of F-actin capable of interaction with myosin and subsequent contraction. The effectiveness of physical agents in producting contraction suggests that the regulation of contraction by the gel is structural in nature.     

The role of polyamines during <Emphasis Type="Italic">in vivo</Emphasis> and <Emphasis Type="Italic">in vitro</Emphasis> development

Polyamines are ubiquitous polycationic compounds that mediate fundamental aspects of cell growth, differentiation, and cell death in eukaryotic and prokaryotic organisms. In plants, polyamines are implicated in a variety of growth and developmental processes, in addition to abiotic and biotic stress responses. In the last decade, mutant studies conducted predominantly in Arabidopsis thaliana revealed an obligatory requirement for polyamines in zygotic and somatic embryogenesis. Moreover, our appreciation for the intricate spatial and temporal regulation of intracellular polyamine levels has advanced considerably. The exact molecular mechanism(s) through which polyamines exert their physiological response remains somewhat enigmatic and likely serves as a major area for future research efforts. In the following review, we discuss recent advances in the plant polyamine field, which range from metabolism and mutant characterization to molecular genetics and potential mode(s) of polyamine action during growth and development in vitro and in vivo. This review will also focus on the specific role of polyamines during embryogenesis and organogenesis.     

Lack of Liver X Receptors Leads to Cell Proliferation in a Model of Mouse Dorsal Prostate Epithelial Cell

Recent studies underline the implication of Liver X Receptors (LXRs) in several prostate diseases such as benign prostatic hyperplasia (BPH) and prostate cancer. In order to understand the molecular mechanisms involved, we derived epithelial cells from dorsal prostate (MPECs) of wild type (WT) or Lxrαβ−/− mice. In the WT MPECs, our results show that LXR activation reduces proliferation and correlates with the modification of the AKT-survival pathway. Moreover, LXRs regulate lipid homeostasis with the regulation of Abca1, Abcg1 and Idol, and, in a lesser extent, Srebp1, Fas and Acc. Conversely cells derived from Lxrαβ−/− mice show a higher basal phosphorylation and consequently activation of the survival/proliferation transduction pathways AKT and MAPK. Altogether, our data point out that the cell model we developed allows deciphering the molecular mechanisms inducing the cell cycle arrest. Besides, we show that activated LXRs regulate AKT and MAPK transduction pathways and demonstrate that LXRs could be good pharmacological targets in prostate disease such as cancer.