Fibroblast dynamics as an in vitro screening platform for anti‐fibrotic drugs in primary myelofibrosis

Although the cause for bone marrow fibrosis in patients with myelofibrosis remains controversial, it has been hypothesized that it is caused by extensive fibroblast proliferation under the influence of cytokines generated by the malignant megakaryocytes. Moreover, there is no known drug therapy which could reverse the process. We studied the fibroblasts in a novel system using the hanging drop method, evaluated whether the fibroblasts obtain from patients are part of the malignant clone of not and, using this system, we screen a large library of FDA‐approved drugs to identify potential drugs candidates that might be useful in the treatment of this disease, specifically which would inhibit fibroblast proliferation and the development of bone marrow fibrosis. We have found that the BM fibroblasts are not part of the malignant clone, as previously suspected and two immunosuppressive medications—cyclosporine and mycophenolate mophetil, as most potent suppressors of the fibroblast collagen production thus potentially inhibitors of bone marrow fibrosis production in myelofibrosis.     

Ancient DNA reveals differences in behaviour and sociality between brown bears and extinct cave bears

Ancient DNA studies have revolutionized the study of extinct species and populations, providing insights on phylogeny, phylogeography, admixture and demographic history. However, inferences on behaviour and sociality have been far less frequent. Here, we investigate the complete mitochondrial genomes of extinct Late Pleistocene cave bears and middle Holocene brown bears that each inhabited multiple geographically proximate caves in northern Spain. In cave bears, we find that, although most caves were occupied simultaneously, each cave almost exclusively contains a unique lineage of closely related haplotypes. This remarkable pattern suggests extreme fidelity to their birth site in cave bears, best described as homing behaviour, and that cave bears formed stable maternal social groups at least for hibernation. In contrast, brown bears do not show any strong association of mitochondrial lineage and cave, suggesting that these two closely related species differed in aspects of their behaviour and sociality. This difference is likely to have contributed to cave bear extinction, which occurred at a time in which competition for caves between bears and humans was likely intense and the ability to rapidly colonize new hibernation sites would have been crucial for the survival of a species so dependent on caves for hibernation as cave bears. Our study demonstrates the potential of ancient DNA to uncover patterns of behaviour and sociality in ancient species and populations, even those that went extinct many tens of thousands of years ago.     

Silencing of cryptic prophages in Corynebacterium glutamicum

DNA of viral origin represents a ubiquitous element of bacterial genomes. Its integration into host regulatory circuits is a pivotal driver of microbial evolution but requires the stringent regulation of phage gene activity. In this study, we describe the nucleoid-associated protein CgpS, which represents an essential protein functioning as a xenogeneic silencer in the Gram-positive Corynebacterium glutamicum. CgpS is encoded by the cryptic prophage CGP3 of the C. glutamicum strain ATCC 13032 and was first identified by DNA affinity chromatography using an early phage promoter of CGP3. Genome-wide profiling of CgpS binding using chromatin affinity purification and sequencing (ChAP-Seq) revealed its association with AT-rich DNA elements, including the entire CGP3 prophage region (187 kbp), as well as several other elements acquired by horizontal gene transfer. Countersilencing of CgpS resulted in a significantly increased induction frequency of the CGP3 prophage. In contrast, a strain lacking the CGP3 prophage was not affected and displayed stable growth. In a bioinformatics approach, cgpS orthologs were identified primarily in actinobacterial genomes as well as several phage and prophage genomes. Sequence analysis of 618 orthologous proteins revealed a strong conservation of the secondary structure, supporting an ancient function of these xenogeneic silencers in phage-host interaction.     

Early Evaluation and Monitoring of Critical Patients with Acute Respiratory Distress Syndrome (ARDS) Using Specific Genetic Polymorphisms

A high percentage of critical patients are found to develop acute respiratory distress syndrome (ARDS). Several studies have reported high mortality rates in these cases which are most frequently associated with multiple organ dysfunctions syndrome. Lately, many efforts have been made to evaluate and monitor ARDS in critical patients. In this regard, the assessment of genetic polymorphisms responsible for developing ARDS present as a challenge and are considered future biomarkers. Early detection of the specific polymorphic gene responsible for ARDS in critically ill patients can prove to be a useful tool in the future, able to help decrease the mortality rates in these cases. Moreover, identifying the genetic polymorphism in these patients can help in the implementation of a personalized intensive therapy scheme for every type of patient, based on its genotype.     

Stabilization of Metals in Subsurface by Biopolymers: Laboratory Drainage Flow Studies

Environmental contamination with heavy metals and radionuclides remains a major problem worldwide. The current clean-up methodologies are based on energy-intensive engineering processes, which are disruptive and costly. A new universal technology targeted for the permanent enclosure and fixation of nuclear and other extreme hazardous metallic wastes in subsurface sites is needed. Such technology will be useful in treating contamination at many sites in the U.S., with specific applications to Department of Energy (DOE) sites. Biopolymers are potential tools for such an innovative technology. Biopolymers have repeated sequences, and therefore provide ample opportunity for chemical reactions with metals, soil particles, and other biopolymers. They also have the additional ability of creating cross-linking interpenetrating networks that can encapsulate the contaminants. Based on this concept, in the present work five biopolymers (xanthan, chitosan, polyhydroxy butyrate, guar gum, polyglutamic acid) were investigated for potential use in the stabilization of metals in the subsurface. The effects of these biopolymers (used alone and in combinations) on soil characteristics (permeability, shear strength) and their metal uptake ability have been studied using laboratory drainage flow systems. Biopolymer solutions were run through the experimental sandpack columns, followed by copper solution and leaching agents (distilled water and hydrochloric acid). The permeability and shear strength of sand were evaluated. Copper uptake capacity of each biopolymer and combination of biopolymers was also studied along with subsequent leaching. All biopolymers tested improved sand characteristics (by decreasing permeability and increasing shear strength) and had good metal uptake ability (60–90%) with relatively low leachability (10–22%). While biopolymers used alone were more efficient in metal uptake, the combination of two biopolymers (xanthan and chitosan) had an increasing plugging effect. These results show the potential of using biopolymers in subsurface metal stabilization.     

Comparative dendroclimatic study of Scots pine, Norway spruce, and silver fir in the Vrancea Range, Eastern Carpathian Mountains

Using dendroclimatical methods, we compared the growth response to climate fluctuations of three of the main Romanian Carpathian Mountains coniferous species, Scots pine (Pinus sylvestris L.), Norway spruce (Picea abies Karst.) and silver fir (Abies alba Mill.), growing intermixed in a unique stand. Climate and soil conditions were therefore the same for all the trees studied. The experimental site was chosen to be representative of the south-exposed sites in the Romanian Carpathian Mountains, where pine occurs naturally. In order to investigate the consequences of fluctuations in climate at different temporal scales, we examined both inter-annual and decadal time-steps. An index of soil water deficit was computed to investigate the consequences of drought. Our study reveals that species exhibited different responses to precipitation, temperature or drought. Overall, pine was the most sensitive to precipitation, while spruce showed a higher response to temperature at high frequency for both the current and the previous growing seasons, and to soil water deficit. Fir was the least sensitive species of the comparison. However, for all species, decadal modulations of growth show precipitation as a common and strong driver on the dry south-facing slopes. The results show that Scots pine would be affected more than fir by increased drought frequency and could in the future be replaced on the xeric sites.     

Predictors of major infections in systemic lupus erythematosus

Introduction  

Infections commonly complicate the course of systemic lupus erythematosus (SLE). Our aim is to investigate the clinical predictors of major infections in patients with SLE.     

Regulation of the Na+/K+ ATPase by Klotho

Klotho-hypomorphic (Klotho(hm)) mice suffer from renal salt wasting and hypovolemia despite hyperaldosteronism. The present study explored the effect of Klotho on renal Na(+)/K(+) ATPase activity. According to immunohistochemistry and confocal microscopy Na(+)/K(+) ATPase protein abundance in isolated collecting ducts was lower in Klotho(hm) mice than in their wild type littermates (Klotho(+/+)). Analysis with dual electrode voltage clamp recording showed that expression of Klotho in Xenopus oocytes increased the Na(+)/K(+) ATPase pump current. Treatment of Xenopus oocytes with Klotho protein similarly increased the pump current. In conclusion, Klotho increases the membrane abundance and activity of the Na(+)/K(+) ATPase. Decreased Na(+)/K(+) ATPase activity could thus contribute to the volume-depletion of klotho(hm) mice.