L-DOPA preloading increases the uptake of borophenylalanine in C6 glioma rat model: a new strategy to improve BNCT efficacy.

Purpose: Boron neutron capture therapy (BNCT) is a radiotherapeutic modality based on 10B(n,a)7Li reaction, for the treatment of malignant gliomas. One of the main limitations for BNCT effectiveness is the insufficient intake of 10B nuclei in the tumor cells. This work was aimed at investigating the use of L-DOPA as a putative enhancer for 10B-drug 4-dihydroxy-borylphenylalanine (BPA) uptake in the C6-glioma model. The investigation was first per- formed in vitro and then extended to the animal model. Methods and Materials: BPA accumulation in C6-glioma cells was assessed using radiowave dielectric spectros- copy, with and without L-DOPA preloading. Two L-DOPA incubation times (2 and 4 hours) were investigated, and the corresponding effects on BPA accumulation were quantified. C6-glioma cells were also implanted in the brain of 32 rats, and tumor growth was monitored by magnetic resonance imaging. Rats were assigned to two experimental branches: (1) BPA administration; (2) BPA administration after pretreatment with L-DOPA. All an- imals were sacrificed, and assessments of BPA concentrations in tumor tissue, normal brain, and blood samples were performed using high-performance liquid chromatography. Results: L-DOPA preloading induced a massive increase of BPA concentration in C6-glioma cells only after a 4-hour incubation. In the animal model, L-DOPA pretreatment produced a significantly higher accumulation of BPA in tumor tissue but not in normal brain and blood samples. Conclusions: This study suggests the potential use of L-DOPA as enhancer for BPA accumulation in malig- nant gliomas eligible for BNCT. L-DOPA preloading effect is discussed in terms of membrane transport mechanisms

From canopy to consumer: what makes and modifes terrestrial DOM in a temperate forest

To investigate how source and processing control the composition of “terrestrial” dissolved organic matter (DOM), we combine soil and tree leachates, tree DOM, laboratory bioincubations, and ultrahigh resolution Fourier-transform ion cyclotron resonance mass spectrometry in three common landscape types (upland forest, forested wetland, and poor fen) of Southeast Alaska’s temperate rainforest. Tree (Tsuga heterophylla and Picea sitchensis) needles and bark and soil layers from each site were leached, and tree stemflow and throughfall collected to examine DOM sources. Dissolved organic carbon concentrations were as high as 167 mg CL−1 for tree DOM, suggesting tree DOM fluxes may be substantial given the hypermaritime climate of the region. Condensed aromatics contributed as much as 38% relative abundance of spruce and hemlock bark leachates suggesting coniferous trees are potential sources of condensed aromatics to surface waters. Soil leachates showed soil wetness dictates DOM composition and processing, with wetland soils producing more aromatic formulae and allowing the preservation of traditionally biolabile, aliphatic formulae. Biodegradation impacted soil and tree DOM differently, and though the majority of source-specific marker formulae were consumed for all sources, some marker formulae persisted. Tree DOM was highly biolabile (> 50%) and showed compositional convergence where processing homogenized DOM from different tree sources. In contrast, wetland and upland soil leachate DOM composition diverged and processing diversified DOM from different soil sources during bioincubations. Increasing precipitation intensity predicted with climate change in Southeast Alaska will increase tree leaching and soil DOM flushing, tightening linkages between terrestrial sources and DOM export to the coastal ocean.The authors thank Emily Whitney for her invaluable feld, laboratory, and logistical assistance and Molly Tankersley for creating Fig. 1. They are also grateful to all the helpful researchers at the National High Magnetic Field Laboratory who enabled data acquisition and processing. This work took place on the lands of the Aak’w Kwáan Tlingit. This work was supported by the National Science Foundation through an NSF Graduate Research Fellowship to MIB. A portion of this work was performed at the National High Magnetic Field Laboratory ICR User Facility, which is supported by the National Science Foundation Division of Chemistry and Division of Materials Research through DMR-1644779 and the State of Florida. Conficts of interest/ Competing interests: The authors have no conficts of interests to declare.Ye

From canopy to consumer: what makes and modifies terrestrial DOM in a temperate forest

To investigate how source and processing control the composition of “terrestrial” dissolved organic matter (DOM), we combine soil and tree leachates, tree DOM, laboratory bioincubations, and ultrahigh resolution Fourier-transform ion cyclotron resonance mass spectrometry in three common landscape types (upland forest, forested wetland, and poor fen) of Southeast Alaska’s temperate rainforest. Tree (Tsuga heterophylla and Picea sitchensis) needles and bark and soil layers from each site were leached, and tree stemflow and throughfall collected to examine DOM sources. Dissolved organic carbon concentrations were as high as 167 mg CL−1 for tree DOM, suggesting tree DOM fluxes may be substantial given the hypermaritime climate of the region. Condensed aromatics contributed as much as 38% relative abundance of spruce and hemlock bark leachates suggesting coniferous trees are potential sources of condensed aromatics to surface waters. Soil leachates showed soil wetness dictates DOM composition and processing, with wetland soils producing more aromatic formulae and allowing the preservation of traditionally biolabile, aliphatic formulae. Biodegradation impacted soil and tree DOM differently, and though the majority of source-specific marker formulae were consumed for all sources, some marker formulae persisted. Tree DOM was highly biolabile (> 50%) and showed compositional convergence where processing homogenized DOM from different tree sources. In contrast, wetland and upland soil leachate DOM composition diverged and processing diversified DOM from different soil sources during bioincubations. Increasing precipitation intensity predicted with climate change in Southeast Alaska will increase tree leaching and soil DOM flushing, tightening linkages between terrestrial sources and DOM export to the coastal ocean.The authors thank Emily Whitney for her invaluable feld, laboratory, and logistical assistance and Molly Tankersley for creating Fig. 1. They are also grateful to all the helpful researchers at the National High Magnetic Field Laboratory who enabled data acquisition and processing. This work took place on the lands of the Aak’w Kwáan Tlingit. This work was supported by the National Science Foundation through an NSF Graduate Research Fellowship to MIB. A portion of this work was performed at the National High Magnetic Field Laboratory ICR User Facility, which is supported by the National Science Foundation Division of Chemistry and Division of Materials Research through DMR-1644779 and the State of Florida. Conficts of interest/ Competing interests: The authors have no conficts of interests to declare.Ye

High-throughput imaging of ATG9A distribution as a diagnostic functional assay for adaptor protein complex 4-associated hereditary spastic paraplegia

Adaptor protein complex 4-associated hereditary spastic paraplegia is caused by biallelic loss-of-function variants in AP4B1, AP4M1, AP4E1 or AP4S1, which constitute the four subunits of this obligate complex. While the diagnosis of adaptor protein complex 4-associated hereditary spastic paraplegia relies on molecular testing, the interpretation of novel missense variants remains challenging. Here, we address this diagnostic gap by using patient-derived fibroblasts to establish a functional assay that measures the subcellular localization of ATG9A, a transmembrane protein that is sorted by adaptor protein complex 4. Using automated high-throughput microscopy, we determine the ratio of the ATG9A fluorescence in the trans-Golgi-network versus cytoplasm and ascertain that this metric meets standards for screening assays (Z'-factor robust >0.3, strictly standardized mean difference >3). The `ATG9A ratio' is increased in fibroblasts of 18 well-characterized adaptor protein complex 4-associated hereditary spastic paraplegia patients [mean: 1.54 +/- 0.13 versus 1.21 +/- 0.05 (standard deviation) in controls] and receiver-operating characteristic analysis demonstrates robust diagnostic power (area under the curve: 0.85, 95% confidence interval: 0.849-0.852). Using fibroblasts from two individuals with atypical clinical features and novel biallelic missense variants of unknown significance in AP4B1, we show that our assay can reliably detect adaptor protein complex 4 function. Our findings establish the 'ATG9A ratio' as a diagnostic marker of adaptor protein complex 4-associated hereditary spastic paraplegia

High-throughput imaging of ATG9A distribution as a diagnostic functional assay for adaptor protein complex 4-associated hereditary spastic paraplegia

Adaptor protein complex 4-associated hereditary spastic paraplegia is caused by biallelic loss-of-function variants in AP4B1, AP4M1, AP4E1 or AP4S1, which constitute the four subunits of this obligate complex. While the diagnosis of adaptor protein complex 4-associated hereditary spastic paraplegia relies on molecular testing, the interpretation of novel missense variants remains challenging. Here, we address this diagnostic gap by using patient-derived fibroblasts to establish a functional assay that measures the subcellular localization of ATG9A, a transmembrane protein that is sorted by adaptor protein complex 4. Using automated high-throughput microscopy, we determine the ratio of the ATG9A fluorescence in the trans-Golgi-network versus cytoplasm and ascertain that this metric meets standards for screening assays (Z'-factor robust >0.3, strictly standardized mean difference >3). The 'ATG9A ratio' is increased in fibroblasts of 18 well-characterized adaptor protein complex 4-associated hereditary spastic paraplegia patients [mean: 1.54 ± 0.13 versus 1.21 ± 0.05 (standard deviation) in controls] and receiver-operating characteristic analysis demonstrates robust diagnostic power (area under the curve: 0.85, 95% confidence interval: 0.849-0.852). Using fibroblasts from two individuals with atypical clinical features and novel biallelic missense variants of unknown significance in AP4B1, we show that our assay can reliably detect adaptor protein complex 4 function. Our findings establish the 'ATG9A ratio' as a diagnostic marker of adaptor protein complex 4-associated hereditary spastic paraplegia

New Materials and Technologies for Durability and Conservation of Building Heritage

The increase in concrete structures’ durability is a milestone to improve the sustainability of buildings and infrastructures. In order to ensure a prolonged service life, it is necessary to detect the deterioration of materials by means of monitoring systems aimed at evaluating not only the penetration of aggressive substances into concrete but also the corrosion of carbon-steel reinforcement. Therefore, proper data collection makes it possible to plan suitable restoration works which can be carried out with traditional or innovative techniques and materials. This work focuses on building heritage and it highlights the most recent findings for the conservation and restoration of reinforced concrete structures and masonry buildings

Deficiency of Antinociception and Excessive Grooming Induced by Acute Immobilization Stress in Per1 Mutant Mice

Acute stressors induce changes in numerous behavioral parameters through activation of the hypothalamic-pituitary-adrenal (HPA) axis. Several important hormones in paraventricular nucleus of the hypothalamus (PVN) play the roles in these stress-induced reactions. Corticotropin-releasing hormone (CRH), arginine-vasopressin (AVP) and corticosterone are considered as molecular markers for stress-induced grooming behavior. Oxytocin in PVN is an essential modulator for stress-induced antinociception. The clock gene, Per1, has been identified as an effecter response to the acute stresses, but its function in neuroendocrine stress systems remains unclear. In the present study we observed the alterations in grooming and nociceptive behaviors induced by acute immobilization stress in Per1 mutant mice and other genotypes (wild types and Per2 mutant). The results displayed that stress elicited a more robust effect on grooming behavior in Per1 mutant mice than in other genotypes. Subsequently, the obvious stress-induced antinociception was observed in the wild-type and Per2 mutant mice, however, in Per1 mutant, this antinociceptive effects were partially-reversed (mechanical sensitivity), or over-reversed to hyperalgesia (thermal sensitivity). The real-time qPCR results showed that in PVN, there were stress-induced up-regulations of Crh, Avp and c-fos in all of genotypes; moreover, the expression change of Crh in Per1 mutant mice was much larger than in others. Another hormonal gene, Oxt, was up-regulated induced by stress in wild-type and Per2 mutant but not in Per1 mutant. In addition, the stress significantly elevated the serum corticosterone levels without genotype-dependent differences, and accordingly the glucocorticoid receptor gene, Nr3c1, expressed with a similar pattern in PVN of all strains. Taken together, the present study indicated that in acute stress treated Per1 mutant mice, there are abnormal hormonal responses in PVN, correlating with the aberrant performance of stress-induced behaviors. Therefore, our findings suggest a novel functional role of Per1 in neuroendocrine stress system, which further participates in analgesic regulation