Replicative Stress Coincides with Impaired Nuclear DNA Damage Response in COX4-1 Deficiency

Cytochrome c oxidase (COX), a multimeric protein complex, is the final electron acceptor in the mitochondrial electron transfer chain. Primary COX deficiency, caused by mutations in either mitochondrial DNA or nuclear-encoded genes, is a heterogenous group of mitochondrial diseases with a wide range of presentations, ranging from fatal infantile to subtler. We previously reported a patient with primary COX deficiency due to a pathogenic variant in COX4I1 (encoding the common isoform of COX subunit 4, COX4-1), who presented with bone marrow failure, genomic instability, and short stature, mimicking Fanconi anemia (FA). In the present study, we demonstrated that accumulative DNA damage coincided primarily with proliferative cells in the patient’s fibroblasts and in COX4i1 knockdown cells. Expression analysis implicated a reduction in DNA damage response pathways, which was verified by demonstrating impaired recovery from genotoxic insult and decreased DNA repair. The premature senescence of the COX4-1-deficient cells prevented us from undertaking additional studies; nevertheless, taken together, our results indicate replicative stress and impaired nuclear DNA damage response in COX4-1 deficiency. Interestingly, our in vitro findings recapitulated the patient’s presentation and present status.     

Screening for the presence of lipophilic marine biotoxins in shellfish samples using the neuro-2a bioassay

The neuro-2a bioassay is considered as one of the most promising cell-based in vitro bioassays for the broad screening of seafood products for the presence of marine biotoxins. The neuro-2a assay has been shown to detect a wide array of toxins like paralytic shellfish poisons (PSPs), ciguatoxins, and also lipophilic marine biotoxins (LMBs). However, the neuro-2a assay is rarely used for routine testing of samples due to matrix effects that, for example, lead to false positives when testing for LMBs. As a result there are only limited data on validation and evaluation of its performance on real samples. In the present study, the standard extraction procedure for LMBs was adjusted by introducing an additional clean-up step with n-hexane. Recovery losses due to this extra step were less than 10%. This wash step was a crucial addition in order to eliminate false-positive outcomes due to matrix effects. Next, the applicability of this assay was assessed by testing a broad range of shellfish samples contaminated with various LMBs, including diarrhetic shellfish toxins/poisons (DSPs). For comparison, the samples were also analysed by LC-MS/MS. Standards of all regulated LMBs were tested, including analogues of some of these toxins. The neuro-2a cells showed good sensitivity towards all compounds. Extracts of 87 samples, both blank and contaminated with various toxins, were tested. The neuro-2a outcomes were in line with those of LC-MS/MS analysis and support the applicability of this assay for the screening of samples for LMBs. However, for use in a daily routine setting, the test might be further improved and we discuss several recommended modifications which should be considered before a full validation is carried out.     

High yield synthesis of boron-based nanosheets

ABSTRACT

We present the chemical exfoliation of magnesium diboride (MgB₂) by employing organic chelating agents for synthesising boron-based nanosheets in high yield. MgB₂ is a promising parent material towards realising quasi-2D forms of boron, owing to its unique layered constitution: graphenic boron planes interleaved with hexagonal arrays of Mg atoms. Chelating agents can selectively extract the interlayer Mg from MgB₂ in water, to form Mg-chelant complexes. The loss of Mg results in delamination of MgB₂ into boron-based nanosheets. This approach is further improved by rational choice of chelants with greater affinity for Mg and tuning the optimal pH for metal complexation, thereby leading to a better yield of nanosheets. These boron-based nanosheets have been explored for their candidacy as a new class of inorganic fillers in a polymeric matrix – polyvinyl alcohol (PVA). The mechanical and thermal properties of these boron-based nanosheet-PVA nanocomposite films have been studied.     

Lithography-based nanoelectrochemistry

Lithographically fabricated nanostructures appear in an increasingly wide range of scientific fields, and electroanalytical chemistry is no exception. This article introduces lithography methods and provides an overview of the new capabilities and electrochemical phenomena that can emerge in nanostructures.     

Repetitive Traumatic Brain Injury Causes Neuroinflammation before Tau Pathology in Adolescent P301S Mice

Repetitive closed head injury (rCHI) is commonly encountered in young athletes engaged in contact and collision sports. Traumatic brain injury (TBI) including rCHI has been reported to be an important risk factor for several tauopathies in studies of adult humans and animals. However, the link between rCHI and the progression of tau pathology in adolescents remains to be elucidated. We evaluated whether rCHI can trigger the initial acceleration of pathological tau in adolescent mice and impact the long-term outcomes post-injury. To this end, we subjected adolescent transgenic mice expressing the P301S tau mutation to mild rCHI and assessed tau hyperphosphorylation, tangle formation, markers of neuroinflammation, and behavioral deficits at 40 days post rCHI. We report that rCHI did not accelerate tau pathology and did not worsen behavioral outcomes compared to control mice. However, rCHI induced cortical and hippocampal microgliosis and corpus callosum astrocytosis in P301S mice by 40 days post-injury. In contrast, we did not find significant microgliosis or astrocytosis after rCHI in age-matched WT mice or sham-injured P301S mice. Our data suggest that neuroinflammation precedes the development of Tau pathology in this rCHI model of adolescent repetitive mild TBI.     

Comparative study of Yb:Lu3Al5O12 and Yb:Lu2O3 laser ceramics produced from laser-ablated nanopowders

We present a comparative study of two Lu-based oxide ceramics doped with Yb³⁺ ions, namely Yb:Lu₃Al₅O₁₂ (garnet) and Yb:Lu₂O₃ (sesquioxide), promising for thin-disk lasers. The ceramics are fabricated using nanopowders of 3.6 at.% Yb:Lu₂O₃ and Al₂O₃ produced by laser ablation: Yb:Lu₃Al₅O₁₂ – by vacuum sintering at 1800 °C for 5 h with the addition of 1 wt% TEOS as a sintering aid, and Yb:Lu₂O₃ – by vacuum pre-sintering at 1250 °C for 2 h followed by Hot Isostatic Pressing at 1400 °C for 2 h under Ar gas pressure of 207 MPa. The comparison includes the structure, Raman spectra, transmission, optical spectroscopy and laser operation. The crystal-field splitting of Yb³⁺ multiplets is revealed for Lu₃Al₅O₁₂. A continuous-wave (CW) Yb:Lu₃Al₅O₁₂ ceramic microchip laser generates 5.65 W at 1031.1 nm with a slope efficiency of 67.2%. In the quasi-CW regime, the peak power is scaled up to 8.83 W. The power scaling for the Yb:Lu₂O₃ ceramic laser is limited by losses originating from residual coloration and inferior thermal behavior.     

A real-time RT-PCR method to detect viable Giardia lamblia cysts in environmental waters

Currently, USEPA Method 1623 is the standard assay used for simultaneous detection of Giardia cysts and Cryptosporidium oocysts in various water matrices. However, the method is unable to distinguish between species, genotype, or to assess viability. Therefore, the objective of the present study was to address the shortcomings of USEPA Method 1623 by developing a novel molecular-based method that can assess viability of Giardia cysts in environmental waters and identify genotypes that pose a human health threat (assemblage groups A and B). Primers and TaqMan^® probes were designed to target the beta-giardin gene in order to discriminate among species and assemblages. Viability was determined by detection of de-novo mRNA synthesis after heat induction. The beta-giardin primer/probe sets were able to detect and differentiate between Giardia lamblia assemblages A and B, and did not detect Giardia muris (mouse species) or G. lamblia assemblages C, D, E and F (non-human), with the exception of Probe A which did detect G. lamblia assemblage F DNA. Additionally, DNA or cDNA of other waterborne organisms were not detected, suggesting that the method is specific to Giardia assemblages. Assay applicability was demonstrated by detection of viable G. lamblia cysts in spiked (assemblage B) and unspiked (assemblage A and B) reclaimed water samples.     

Perchlorate exposure in lactating women in an urban community in New Jersey

Perchlorate is most widely known as a solid oxidant for missile and rocket propulsion systems although it is also present as a trace contaminant in some fertilizers. It has been detected in drinking water, fruits, and vegetables throughout New Jersey and most of the United States. At sufficiently high doses, perchlorate interferes with the uptake of iodine into the thyroid and may interfere with the development of the skeletal system and the central nervous system of infants. Therefore, it is important to quantify perchlorate in breast milk to understand potential perchlorate exposure in infants. In this study we measured perchlorate in breast milk, urine, and drinking water collected from 106 lactating mothers from Central New Jersey. Each subject was asked to provide three sets of samples over a 3-month period. The average ± SD perchlorate level in drinking water, breast milk, and urine was 0.168 ± 0.132 ng/mL (n = 253), 6.80 ± 8.76 ng/mL (n = 276), and 3.19 ± 3.64 ng/mL (3.51 ± 6.79 μg/g creatinine) (n = 273), respectively. Urinary perchlorate levels were lower than reference range values for women of reproductive age (5.16 ± 11.33 μg/g creatinine, p = 0.03), likely because of perchlorate secretion in breast milk. Drinking water perchlorate levels were ≤ 1.05 ng/mL and were not positively correlated with either breast milk or urine perchlorate levels. These findings together suggest that drinking water was not the most important perchlorate exposure source for these women. Creatinine-adjusted urine perchlorate levels were strongly correlated with breast milk perchlorate levels (r = 0.626, p = < 0.0005). Breast milk perchlorate levels in this study are consistent with widespread perchlorate exposure in lactating women and thus infants. This suggests that breast milk may be a source of exposure to perchlorate in infants.