Protocolized Brain Oxygen Optimization in Subarachnoid Hemorrhage.

Brain tissue hypoxia (P <sub>bt</sub> O <sub>2</sub> < 20 mmHg) is common after subarachnoid hemorrhage (SAH) and associated with poor outcome. Recent data suggest that brain oxygen optimization is feasible and reduces the time spent with P <sub>bt</sub> O <sub>2</sub> < 20 mmHg from 45 to 16% in patients with severe traumatic brain injury. Here, we intended to quantify the brain tissue hypoxia burden despite implementation of a protocolized treatment approach in poor-grade SAH patients and to identify the simultaneous occurrence of pathologic values potentially amenable to treatment. We present a bi-centric observational cohort study including 100 poor-grade SAH patients admitted to two tertiary care centers who underwent multimodal brain monitoring and were managed with a P <sub>bt</sub> O <sub>2</sub> -targeted protocolized approach. P <sub>bt</sub> O <sub>2</sub> optimization (≥ 20 mmHg) included a stepwise neuro-intensive care approach, aiming to prevent low cerebral perfusion pressure (CPP), and blood hemoglobin, and to keep normocapnia, normoxemia, and normothermia. Based on routine blood gas analysis, hemoglobin, PaCO <sub>2,</sub> and PaO <sub>2</sub> data were matched to 2-h averaged data of continuous CPP, P <sub>bt</sub> O <sub>2</sub> , core temperature, and to hourly cerebral microdialysis (CMD) samples over the first 11 days. Patients had a Glasgow Coma Scale of 3 (IQR 3-4) and were 58 years old (IQR 48-66). Overall incidence of brain tissue hypoxia was 25%, which was not different between both sites despite differences in the treatment approach. During brain tissue hypoxia, episodes of CPP < 70 mmHg (27%), PaCO <sub>2</sub> < 35 mmHg (19%), PaO <sub>2</sub> < 80 mmHg (14%), Hb < 9 g/dL (11%), metabolic crisis (CMD-lactate/pyruvate ratio > 40, and CMD-glucose < 0.7 mmol/L; 7%), and temperature > 38.3 °C (4%) were common. Our results demonstrate that brain tissue hypoxia remains common despite implementation of a P <sub>bt</sub> O <sub>2</sub> -targeted therapy in poor-grade SAH patients, suggesting room for further optimization

Effects of Zero Tillage (No-Till) Conservation Agriculture on soil physical and biological properties and their contributions to sustainability.

Not cultivating soil, rotating crops over the years, and leaving crop residues on the surface in the practice of zero tillage/conservation agriculture (ZT/CA) reverses the historically accelerating degradation of soil organic matter (SOM) and soil structure, while increasing soil biological activity by a factor of 2 to 4. The results of this are many: (a) not cultivating reduces soil compaction, leaving old root holes to facilitate internal drainage, averts the pulverization of soil aggregates and formation of pans, reduces draft power for planting and gives shelter, winter food and nesting sites for fauna, (b) crop residues on the surface practically eliminate wind and water erosion, reduce soil moisture loss through the mulch effect, slow spring warm-up (possibly offset by a lower specific heat demand with less water retention in surface soil) and act as a reserve of organically-compounded nutrients (as they decompose to humus), (c) more SOM means higher available water and nutrient retention, higher biological activity year round (enhancing biological controls), higher levels of water-stable aggregates and a positive carbon sink in incremental SOM. The positive impacts for society are: (i) more and cheaper food, (ii) reduced flood and drought-induced famine risks, (iii) a positive carbon sink in SOM and possible reductions in NO emissions, (iv) cleaner water and greater aquifer recharge due to reduced runoff, (v) cleaner air through effective elimination of dust as a product of cultivation (vi) less water pollution and greater aquifer recharge from reduced rainfall runoff, (vii) farm diesel consumption halved, (viii) reduced demand for (tropical) de-forestation, by permitting crop expansion on steeper lands, (ix) increased wildlife populations (skylarks, plovers, partridge and peccaries) and (x) an improved conservation mindset in farmers. It is notable that, in spite of successful practitioners in all European countries, mainstream adoption is still to come: Europe s ZT/CA area is 1.35 million hectares, while the world area is now some 125 million and growing at a rate of 7 million hectares per year. More scientific measurements of the benefits of this system are required, both to assist adoption and to trigger policy measures. In the EEC, CAP reform (greening) needs to consider making environmental services payments for these social benefits since a reduction in single farm payments is ineluctable and carbon footprint reduction is of the essence, in the face of constantly-rising fuel prices and the need to cut GHG emissions. Therefore, as the principal farm tool which offers an effective and immediate solution towards positive changes in soil quality, productivity and sustainability, ZT/CA adoption needs financial incentives, which have high economic and environmental returns to society

Association between intra-radicular posts and periapical lesions in endodontically treated teeth

Introduction: A significant number of endodontically treated teeth restored with posts have associated periapical lesions, and several authors have discussed the probable causes of the development of these. Attention has been focused on restorative procedures performed after endodontic treatment and their association with the prognosis of endodontic therapy because a number of root-filled teeth will require post- and core-retained restorations. Purpose: The purpose of this study was to evaluate, by examination of periapical radiographs, whether the placement of intra-radicular posts in endodontically treated teeth may act as a risk factor for development of periapical lesions. Material and Methods: This case-control study analyzed periapical radiographs of 72 endodontically treated teeth with coronal restorations. All radiographs were obtained from a single private practice. Specimens were assigned to 2 groups: Group 1 (control) was composed of teeth without periapical lesions and Group 2 (case) was composed of teeth with periapical lesions. The number of teeth with and without posts in each group was recorded. Three calibrated examiners analyzed the radiographs visually under X4 magnification. Results: In Group 1, 28 (65.1%) out of 43 teeth were restored with posts. In Group 2, 24 (82.8%) out of 29 teeth had intra-radicular posts. The interpretation of chi-square test showed that these percentages were not significantly different (x²=2.687; p=0.101). Odds ratio was 2.571 (0.815-8.118), which indicates that there was no statistically significant association between periapical lesions and posts. Conclusion: Intra-radicular posts placed in endodontically treated teeth were not a significant risk factor for development of periapical lesions in the practice where the cohort of patients was treated

LRF maintains genome integrity by regulating the non-homologous end joining pathway of DNA repair

Leukemia/lymphoma-related factor (LRF) is a POZ/BTB and Krüppel (POK) transcriptional repressor characterized by context-dependent key roles in cell fate decision and tumorigenesis. Here we demonstrate an unexpected transcription-independent function for LRF in the classical non-homologous end joining (cNHEJ) pathway of double-strand break (DSB) repair. We find that LRF loss in cell lines and mouse tissues results in defective cNHEJ, genomic instability and hypersensitivity to ionizing radiation. Mechanistically, we show that LRF binds and stabilizes DNA-PKcs on DSBs, in turn favouring DNA-PK activity. Importantly, LRF loss restores ionizing radiation sensitivity to p53 null cells, making LRF an attractive biomarker to direct p53-null LRF-deficient tumours towards therapeutic treatments based on genotoxic agents or PARP inhibitors following a synthetic lethal strategy

Cell Cycle-Dependent Induction of Homologous Recombination by a Tightly Regulated I-SceI Fusion Protein

Double-strand break repair is executed by two major repair pathways: non-homologous end joining (NHEJ) and homologous recombination (HR). Whereas NHEJ contributes to the repair of ionizing radiation (IR)-induced double strand breaks (DSBs) throughout the cell cycle, HR acts predominantly during the S and G2 phases of the cell cycle. The rare-cutting restriction endonuclease, I-SceI, is in common use to study the repair of site-specific chromosomal DSBs in vertebrate cells. To facilitate analysis of I-SceI-induced DSB repair, we have developed a stably expressed I-SceI fusion protein that enables precise temporal control of I-SceI activation, and correspondingly tight control of the timing of onset of site-specific chromosome breakage. I-SceI-induced HR showed a strong, positive linear correlation with the percentage of cells in S phase, and was negatively correlated with the G1 fraction. Acute depletion of BRCA1, a key regulator of HR, disrupted the relationship between S phase fraction and I-SceI-induced HR, consistent with the hypothesis that BRCA1 regulates HR during S phase

High-throughput miRNA profiling of human melanoma blood samples

<p>Abstract</p> <p>Background</p> <p>MicroRNA (miRNA) signatures are not only found in cancer tissue but also in blood of cancer patients. Specifically, miRNA detection in blood offers the prospect of a non-invasive analysis tool.</p> <p>Methods</p> <p>Using a microarray based approach we screened almost 900 human miRNAs to detect miRNAs that are deregulated in their expression in blood cells of melanoma patients. We analyzed 55 blood samples, including 20 samples of healthy individuals, 24 samples of melanoma patients as test set, and 11 samples of melanoma patients as independent validation set.</p> <p>Results</p> <p>A hypothesis test based approch detected 51 differentially regulated miRNAs, including 21 miRNAs that were downregulated in blood cells of melanoma patients and 30 miRNAs that were upregulated in blood cells of melanoma patients as compared to blood cells of healthy controls. The tets set and the independent validation set of the melanoma samples showed a high correlation of fold changes (0.81). Applying hierarchical clustering and principal component analysis we found that blood samples of melanoma patients and healthy individuals can be well differentiated from each other based on miRNA expression analysis. Using a subset of 16 significant deregulated miRNAs, we were able to reach a classification accuracy of 97.4%, a specificity of 95% and a sensitivity of 98.9% by supervised analysis. MiRNA microarray data were validated by qRT-PCR.</p> <p>Conclusions</p> <p>Our study provides strong evidence for miRNA expression signatures of blood cells as useful biomarkers for melanoma.</p

A Novel 3-Hydroxysteroid Dehydrogenase That Regulates Reproductive Development and Longevity

A multidisciplinary approach identifies novel biochemical activities involved in the synthesisof C. elegans bile acid-like steroids, which act as hormones that regulate sterol metabolism and longevity

Rif1 S-acylation mediates DNA double-strand break repair at the inner nuclear membrane

Rif1 is involved in telomere homeostasis, DNA replication timing, and DNA double-strand break (DSB) repair pathway choice from yeast to human. The molecular mechanisms that enable Rif1 to fulfill its diverse roles remain to be determined. Here, we demonstrate that Rif1 is S-acylated within its conserved N-terminal domain at cysteine residues C466 and C473 by the DHHC family palmitoyl acyltransferase Pfa4. Rif1 S-acylation facilitates the accumulation of Rif1 at DSBs, the attenuation of DNA end-resection, and DSB repair by non-homologous end-joining (NHEJ). These findings identify S-acylation as a posttranslational modification regulating DNA repair. S-acylated Rif1 mounts a localized DNA-damage response proximal to the inner nuclear membrane, revealing a mechanism of compartmentalized DSB repair pathway choice by sequestration of a fatty acylated repair factor at the inner nuclear membrane

Replication intermediates that escape Dna2 activity are processed by Holliday junction resolvase Yen1

Cells have evolved mechanisms to protect, restart and repair perturbed replication forks, allowing full genome duplication, even under replication stress. Interrogating the interplay between nuclease-helicase Dna2 and Holliday junction (HJ) resolvase Yen1, we find the Dna2 helicase activity acts parallel to homologous recombination (HR) in promoting DNA replication and chromosome detachment at mitosis after replication fork stalling. Yen1, but not the HJ resolvases Slx1-Slx4 and Mus81-Mms4, safeguards chromosome segregation by removing replication intermediates that escape Dna2. Post-replicative DNA damage checkpoint activation in Dna2 helicase-defective cells causes terminal G2/M arrest by precluding Yen1-dependent repair, whose activation requires progression into anaphase. These findings explain the exquisite replication stress sensitivity of Dna2 helicase-defective cells, and identify a non-canonical role for Yen1 in the processing of replication intermediates that is distinct from HJ resolution. The involvement of Dna2 helicase activity in completing replication may have implications for DNA2-associated pathologies, including cancer and Seckel syndrome