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Objective: Indigenous peoples in Canada are considered the highest-risk populations for type 2 diabetes mellitus (T2DM). Quality improvement (QI) strategies may be considered effective interventions to improving T2DM health outcomes. The purpose of this study was to understand experiences associated with the implementation of the SOAR QI program to improve prevention and management of T2DM., Method: A qualitative study was conducted, and in-depth, semi-structured interviews were held with QI team members and key contacts, in person and through videoconference with two First Nations communities. Interviews were audio recorded and transcribed for data analysis., Results: 10 interviews were conducted and emerging themes from the data analysis were organized into two categories, namely facilitators and barriers. Four subthemes were identified. Two subthemes emerged under the category of facilitators (cultural relevance, and partnership building). Two subthemes emerged under the category of barriers, (workload burden, role ambiguity)., Discussion: This study highlighted the necessity of implementing diabetes QI strategies that foster cultural sensitivity and provide opportunities for partnership building, to strengthen community relationships. The study also highlighted the importance of diminishing role ambiguity, and increased workload burdens, which can hinder the successful implementation of QI programs long-term., Conclusion: The findings of this study can be utilized to improve future adaptations of SOAR and other diabetes First Nations focused QI strategies, to benefit Indigenous people in acquiring optimal outcomes relative to T2DM care. Findings can also inform the design, practices, and policies of such QI interventions in support of the spread and sustainability of the intervention long-term., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Women are the fastest growing population among Veterans and have substantial risk factors that increase their likelihood for developing cancer. To ensure that the Department of Veterans Affairs Veterans Health Administration (VHA) offers the best possible cancer care to women Veterans, it established the Breast and Gynecologic Oncology System of Excellence (BGSoE) in 2021. The BGSoE offers telehealth oncology services and a comprehensive cancer navigation program. Veterans are identified through physician referral or through the BGSoE dashboard which integrates ICD-10 codes and text mining from VA electronic health records to identify eligible Veterans with breast or gynecological cancers. Descriptive statistics, including Veteran demographics and geographical location, were derived from BGSoE dashboard data. From January 1, 2021 to March 15, 2024, the BGSoE identified a total of 7,187 incident cases of breast or gynecological cancer among living Veterans. Most cancers were breast (78%) versus gynecological cancers (22%) and 10% of Veterans with breast cancer were identified as male. The average age at diagnosis was 59 for Veterans with breast cancer and 56 for those Veterans with gynecological cancers. Among Veterans in the BGSoE, 28% identified as Black and 6% identified as Hispanic. As the prevalence of women Veterans requiring cancer-related care continues to rise, it will be essential for VHA to evaluate the equitable reach, quality, and acceptability of women-focused cancer health services. The BGSoE focuses on providing high-quality and coordinated clinical cancer care. Veterans Health Administration also established the Center for Oncology Outcomes Review and Gender (COURAGE) to evaluate the BGSoE and continue to strengthen cancer care services in VHA. Initial evaluation objectives include establishing an evidence base regarding Veterans with breast and gynecological cancers, including their experiences with cancer care in the VHA. Eventually, COURAGE will provide ongoing monitoring and evaluation to continue to grow and improve cancer care in the VHA., (Published by Oxford University Press on behalf of the Association of Military Surgeons of the United States 2024. This work is written by (a) US Government employee(s) and is in the public domain in the US.)

Background: Women are the fastest growing veteran group in the US and the number of women veterans (WVs) with cancer is rising; however, little is known about this population. Cancer care for WVs is complex and it is essential to understand their unique needs and care coordination challenges to provide evidence-based care. The purpose of this review is to map the quantity, distribution, and characteristics of literature describing cancer and its treatment among WVs., Methods: We searched MEDLINE (via PubMed), Embase (Elsevier), and Web of Science Core Collection (Clarivate) from inception through January, 2024. Publications were eligible that reported gender-specific data on any aspect of cancer care among WVs. Data was abstracted by a single investigator with over-reading., Results: Forty-six reports were included; 44 were observational and 19 had a women-only sample. There were no interventional reports and no qualitative reports had a patient sample. Breast cancer was the most commonly addressed (n = 19). There were six additional reports on sex-specific cancers. Many reports used large VA databases or previous trial data, creating the potential for patient overlap between reports. Among VA-specific areas of interest, only three reports evaluated the potential implications of racial differences and only two included a transgender population. No reports examined the effects of toxic exposures on cancer. Within the NCI Cancer Control Continuum, crosscutting areas were more commonly represented; over half (25) of the reports addressed epidemiology. There were few reports on focus areas and little overlap between focus and crosscutting areas., Discussion: Existing literature provides an inadequate understanding of the population of WVs with cancer. There is scant information regarding the population of WVs with cancer, their care preferences or experiences, or how to best identify and address unmet healthcare needs. It is imperative to expand research to provide evidence-based care for this population., (© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

Background: The aim of this case report was to present a translational approach to tooth autotransplantation using jiggling forces to enlarge the periodontal ligament (PDL) space before autotransplantation, with the goal of improving treatment success and long-term survival., Methods: A 23-year-old patient, undergoing orthodontic therapy and with an unrestorable maxillary first molar, was proposed to have a healthy and fully-erupted maxillary third molar transplanted in the socket of the first molar. Jiggling forces were applied to the third molar to enlarge the PDL space and facilitate the preservation of PDL fibers on the root surfaces during the extraction., Results: Jiggling forces induced hypermobility and widened PDL space of the third molar. The autotransplantation was successful and the patient was followed regularly over a 27-year period. At the 27-year visit, the patient showed optimal chewing function, oral plaque control, and absence of gingivitis. The transplanted molar exhibited periodontal health and absence of mobility. Probing depth of 5 mm and radiographic external root resorption was noted on a localized area of the transplanted tooth which had experienced traumatic and unintentional removal of PDL fibers during the extraction., Conclusions: A translational approach was proposed by integrating knowledge from the fields of orthodontics, trauma from occlusion, and replantation. It validated the crucial importance of maintaining healthy PDL fibers on the root surface and demonstrated clinically the successful autotransplantation of a fully formed third molar into the socket of a first molar with a retention of 27 years., Key Points: Why is this case new information? This case provided evidence of successful autotransplantation of a molar with complete root formation. It reported the longest-term follow-up (27 years) present in the literature. Most importantly, it used a translational medicine approach to apply concepts from the fields of orthodontics and traumatic occlusion to improve the success of the autotransplantation procedure. What are the keys to the successful management of this case? Jiggling forces induced tooth hypermobility and increased the PDL space of the tooth planned for autotransplantation. In turn, they facilitated the atraumatic extraction and preservation of the PDL fibers on the transplanted tooth, improving the success of the reattachment of periodontal fibers. What are the primary limitations to success in this case? Traumatic extraction resulting in the unintended removal of PDL fibers from the tooth planned for autotransplantation, or intentional removal of PDL fibers with root planing are expected to decrease the success rate of the autotransplantation procedure. This is due to the lack of viable PDL cells necessary for reattachment., (© 2023 American Academy of Periodontology.)

Programmed ribosomal frameshifting is a translational recoding phenomenon in which a proportion of ribosomes are stimulated to slip backwards or forwards on an mRNA 1 , rephasing the ribosome relative to the mRNA. While frameshifting is often employed by viruses 2 , very few phylogenetically conserved examples are known in vertebrate genes and the evidence for some of these is controversial 3,4 . Here we report a +1 frameshifting signal in the coding sequence of the human gene PLEKHM2 , encoding the ARL8-dependent, lysosome-kinesin-1 adaptor protein PLEKHM2 5 . This +1 frameshifting signal, UCC_UUU_CGG, is highly conserved in vertebrates and exhibits an influenza virus-like frameshift motif with similar efficiency 6,7 . Purification and mass spectrometry of GFP-tagged trans-frame protein from cells confirms frameshifting. Structure prediction shows that the new C-terminal domain generated by this frameshift forms an alpha-helix. This additional domain relieves PLEKHM2 from autoinhibition, allowing it to move to the tips of cells via association with kinesin-1 without requiring activation by ARL8. Thus, the frameshift proteoform generates a constitutively active adaptor of kinesin-1.

Competing Interests: Declaration of competing interest Rosemarie Walch, DO. Robert Pace, MD. Jeanie Cote MD: speakers bureau/consultant/advisory boards for Alexion, Amgen/Horizon, Biogen, EMD Serono, Genentech, Novartis, Sanofi, and TG therapeutics Paige Goulette, DNP: consultant/advisory board for Amgen/Horizon, EMD Serono, Sanofi and TG therapeutics Molly Davis-Yerian, BA: advisory board for TG therapeutics Sam Katt, RN: no disclosures Institution of all (Memorial Healthcare) Research funding from: Alexion, Amgen/Horizon, Biogen, EMD Serono Genentech, Novartis, Sanofi and TG therapeutics

Background: This study presents the diagnois, management, and tissue response to an acute periodontal lesion with deep pocketing affecting a maxillary central incisor in a young patient devoid of caries or a history of periodontitis., Methods: Clinical and radiographic examinations facilitated the diagnosis of the pathology as an endoperiodontal lesion (EPL) with root damage, exhibiting supracrestal invasive root resorption. Orthograde endodontic therapy was employed to decontaminate and seal the endodontic space. The resorptive site was treated through the endodontic access, debrided, and sealed. No periodontal therapy (surgical or nonsurgical) was performed. No mechanical instrumentation was performed within the pocketed root surface., Results: At 6-month and 1-year follow-ups after endodontic therapy the periodontium displayed a physiologically healthy condition without pus or inflammation, exhibiting a circumferential probing depth of 2 mm, and absence of tooth mobility. These favorable outcomes persisted throughout a 4-year follow-up period., Conclusions: The spontaneous healing of pocketing and abscess occurred without mechanical root instrumentation following endodontic therapy and treatment of external invasive root resorption in an EPL., Key Points: Accurate diagnosis and identification of relevant etiologic factors are pivotal for effectively managing endodontic-periodontal lesions. Once a diagnosis is established, the therapy focuses on eliminating the primary cause, followed by a subsequent diagnostic phase after healing. The definitive understanding of the diagnosis and etiology of endodontic-periodontal lesions often becomes clear in retrospect, based on the outcomes of the therapy. When probing acute periodontal lesions, deep probing depths may occur without permanent loss of periodontal attachment. If the acute lesion was not induced by a periodontal cause and if no periodontal etiology arises secondarily, resolving the primary cause of the endoperiodontal lesion can lead to the spontaneous resolution of the pocketing. This results in spontaneous healing of periodontium without the need for intentional periodontal therapy. A clinical dilemma arises when considering periodontal treatment during the acute inflammatory phase of endo-periodontal pathology. It is advisable to refrain from mechanical root instrumentation particularly if a clear periodontal cause is not apparent, to prevent from iatrogenic damage to periodontal fibers and the potential risk of gingival recessions. However, this does not imply avoiding periodontal therapy entirely for every case. Rather, it is recommended to delay the decision on root instrumentation until a new diagnostic phase is conducted following the healing of the endodontic etiology., (© 2024 The Authors. Clinical Advances in Periodontics published by Wiley Periodicals LLC on behalf of American Academy of Periodontology.)

In neurons, RNA granules are transported along the axon for local translation away from the soma. Recent studies indicate that some of this transport involves hitchhiking of RNA granules on lysosome-related vesicles. In the present study, we leveraged the ability to prevent transport of these vesicles into the axon by knockout of the lysosome-kinesin adaptor BLOC-one-related complex (BORC) to identify a subset of axonal mRNAs that depend on lysosome-related vesicles for transport. We found that BORC knockout causes depletion of a large group of axonal mRNAs mainly encoding ribosomal and mitochondrial/oxidative phosphorylation proteins. This depletion results in mitochondrial defects and eventually leads to axonal degeneration in human induced pluripotent stem cell (iPSC)-derived and mouse neurons. Pathway analyses of the depleted mRNAs revealed a mechanistic connection of BORC deficiency with common neurodegenerative disorders. These results demonstrate that mRNA transport on lysosome-related vesicles is critical for the maintenance of axonal homeostasis and that its failure causes axonal degeneration., (© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

BLOC-one-related complex (BORC) is a multiprotein complex composed of eight subunits named BORCS1-8. BORC associates with the cytosolic face of lysosomes, where it sequentially recruits the small GTPase ARL8 and kinesin-1 and -3 microtubule motors to promote anterograde transport of lysosomes toward the peripheral cytoplasm in non-neuronal cells and the distal axon in neurons. The physiological and pathological importance of BORC in humans, however, remains to be determined. Here, we report the identification of compound heterozygous variants [missense c.85T>C (p.Ser29Pro) and frameshift c.71-75dupTGGCC (p.Asn26Trpfs*51)] and homozygous variants [missense c.196A>C (p.Thr66Pro) and c.124T>C (p.Ser42Pro)] in BORCS8 in five children with a severe early-infantile neurodegenerative disorder from three unrelated families. The children exhibit global developmental delay, severe-to-profound intellectual disability, hypotonia, limb spasticity, muscle wasting, dysmorphic facies, optic atrophy, leuko-axonopathy with hypomyelination, and neurodegenerative features with prevalent supratentorial involvement. Cellular studies using a heterologous transfection system show that the BORCS8 missense variants p.Ser29Pro, p.Ser42Pro and p.Thr66Pro are expressed at normal levels but exhibit reduced assembly with other BORC subunits and reduced ability to drive lysosome distribution toward the cell periphery. The BORCS8 frameshift variant p.Asn26Trpfs*51, on the other hand, is expressed at lower levels and is completely incapable of assembling with other BORC subunits and promoting lysosome distribution toward the cell periphery. Therefore, all the BORCS8 variants are partial or total loss-of-function alleles and are thus likely pathogenic. Knockout of the orthologous borcs8 in zebrafish causes decreased brain and eye size, neuromuscular anomalies and impaired locomotion, recapitulating some of the key traits of the human disease. These findings thus identify BORCS8 as a novel genetic locus for an early-infantile neurodegenerative disorder and highlight the critical importance of BORC and lysosome dynamics for the development and function of the central nervous system., (Published by Oxford University Press on behalf of the Guarantors of Brain 2023.)

Osteosarcoma (OS) cancer treatments include systemic chemotherapy and surgical resection. In the last years, novel treatment approaches have been proposed, which employ a drug-delivery system to prevent offside effects and improves treatment efficacy. Locally delivering anticancer compounds improves on high local concentrations with more efficient tumour-killing effect, reduced drugs resistance and confined systemic effects. Here, the synthesis of injectable strontium-doped calcium phosphate (SrCPC) scaffold was proposed as drug delivery system to combine bone tissue regeneration and anticancer treatment by controlled release of methotrexate (MTX) and doxorubicin (DOX), coded as SrCPC-MTX and SrCPC-DOX, respectively. The drug-loaded cements were tested in an in vitro model of human OS cell line SAOS-2, engineered OS cell line (SAOS-2-eGFP) and U2-OS. The ability of doped scaffolds to induce OS cell death and apoptosis was assessed analysing cell proliferation and Caspase-3/7 activities, respectively. To determine if OS cells grown on doped-scaffolds change their migratory ability and invasiveness, a wound-healing assay was performed. In addition, the osteogenic potential of SrCPC material was evaluated using human adipose derived-mesenchymal stem cells. Osteogenic markers such as (i) the mineral matrix deposition was analysed by alizarin red staining; (ii) the osteocalcin (OCN) protein expression was investigated by enzyme-linked immunosorbent assay test, and (iii) the osteogenic process was studied by real-time polymerase chain reaction array. The delivery system induced cell-killing cytotoxic effects and apoptosis in OS cell lines up to Day 7. SrCPC demonstrates a good cytocompatibility and it induced upregulation of osteogenic genes involved in the skeletal development pathway, together with OCN protein expression and mineral matrix deposition. The proposed approach, based on the local, sustained release of anticancer drugs from nanostructured biomimetic drug-loaded cements is promising for future therapies aiming to combine bone regeneration and anticancer local therapy., (© 2024 The Authors. Journal of Cellular Physiology published by Wiley Periodicals LLC.)

Adaptor protein complex 4 (AP-4) is a heterotetrameric complex that promotes export of selected cargo proteins from the trans-Golgi network. Mutations in each of the AP-4 subunits cause a complicated form of Hereditary Spastic Paraplegia (HSP). Herein, we report that ApoER2, a receptor in the Reelin signaling pathway, is a cargo of the AP-4 complex. We identify the motif ISSF/Y within the ApoER2 cytosolic domain as necessary for interaction with the canonical signal-binding pocket of the µ4 (AP4M1) subunit of AP-4. AP4E1- knock-out (KO) HeLa cells and hippocampal neurons from Ap4e1-KO mice display increased co-localization of ApoER2 with Golgi markers. Furthermore, hippocampal neurons from Ap4e1-KO mice and AP4M1-KO human iPSC-derived cortical i3Neurons exhibit reduced ApoER2 protein expression. Analyses of biosynthetic transport of ApoER2 reveal differential post-Golgi trafficking of the receptor, with lower axonal distribution in KO compared to wild-type neurons, indicating a role of AP-4 and the ISSF/Y motif in the axonal localization of ApoER2. Finally, analyses of Reelin signaling in mouse hippocampal and human cortical KO neurons show that AP4 deficiency causes no changes in Reelin-dependent activation of the AKT pathway and only mild changes in Reelin-induced dendritic arborization, but reduces Reelin-induced ERK phosphorylation, CREB activation, and Golgi deployment. This work thus establishes ApoER2 as a novel cargo of the AP-4 complex, suggesting that defects in the trafficking of this receptor and in the Reelin signaling pathway could contribute to the pathogenesis of HSP caused by mutations in AP-4 subunits., Competing Interests: Declaration of Competing Interest The authors state that this research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Nature-based solutions inherently require a multifaceted perspective that encompasses diverse fields. The aim of this project is to develop more effective nature-based solutions, climate action and environmental awareness by breaking down boundaries between disciplines and fostering a co-creative process. Concepts of ecology and urban forestry were combined with the research on political ecology, environmental humanities, land art, regenerative art, performing art, participatory art, and more-than-human art. This process resulted in the creation of Aula Verde Aniene. It is located in an urban park in Rome and consists of a stand of trees arranged in circles with a specific design to give the perception of being in an outdoor vegetated room. The project activities involved community participation through art performances and citizen science initiatives. Regulating and cultural ecosystem services of Aula Verde were assessed using i-Tree Eco software and citizens' surveys. Beyond numerical descriptions of ecosystem services, the manuscript introduces shinrin-yoku as a practice to raise awareness of nature. The distinctive approach here described contributed to convey a sense of belonging to the ecosystem to citizens. The project framework and study findings have been developed to formulate policy recommendations and disseminate a format that can be adapted to diverse locations., (© 2024. The Author(s).)

Objective: To reduce liver and lung dose during right breast irradiation while maintaining optimal dose to the target volume. This dose reduction has the potential to decrease acute side effects and long-term toxicity., Materials and Methods: 16 patients treated with radiation therapy for localized carcinoma of the right breast were included retrospectively. For the planning CT, each patient was immobilised on an indexed board with the arms placed above the head. CT scans were acquired in free-breathing (FB) as well as with deep inspiration breath hold (DIBH). Both scans were acquired with the same length. Planning target volumes (PTV's) were created with a 5 mm margin from the respective clinical target volumes (CTV's) on both CT datasets. The liver was outlined as scanned. Dose metrics evaluated were as follows: differences in PTV coverage, dose to the liver (max, mean, V90%, V50%, V30%), dose to lung (mean, V20Gy, relative electron density) and dose to heart (Dmax). The p-values were calculated using Wilcoxon signed-rank tests. A p-value was significant when <0.05., Results: Differences in PTV coverage between plans using FB and DIBH were less than 2 %. Maximum liver dose was significantly less using DIBH: 17.5 Gy versus FB: 40.3 Gy (p < 0.001). The volume of the liver receiving 10 % of the dose was significantly less using DIBH with 1.88 cm 3 versus 72.2 cm 3 under FB (p < 0.001). The absolute volume receiving 20 Gy in the right lung was larger using DIBH: 291 cm 3 versus 230 cm 3 under FB (p < 0.001) and the relative volume of lung receiving dose greater than 20 Gy was smaller with DIBH: 11.5 % versus 14 % in FB (p = 0.007). The relative electron density of lung was significantly less with DIBH: 0.59 versus 0.62 with FB, (p < 0.001). This suggests that the lung receives less dose due to its lower density when using DIBH., Conclusion: Radiation of the right breast using DIBH spares liver and lung tissue significantly and thus carries the potential of best practice for right sided breast cancer., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 The Authors.)

Introduction: Human polyomaviruses (HPyVs) cause persistent/latent infections in a large fraction of the population. HPyV infections may cause severe diseases in immunocompromised patients. Malawi polyomavirus (MWPyV) is the 10th discovered human polyomavirus (HPyV 10). MWPyV was found in stool samples of healthy children. So far, the few investigations carried out on HPyV 10 did not find an association with human disease., Methods: In this study, to verify the putative association between MWPyV and human diseases, MWPyV seroprevalence was investigated in patients affected by i) lymphoproliferative disorders (LPDs) and ii) immune system disorders, i.e., autoimmune diseases (ADs), and in iii) healthy subjects. An indirect ELISA, employing virus-like particles (VLPs) to detect serum IgG antibodies against MWPyV/HPyV 10, was carried out. The study also revealed the prevalence of another polyomavirus, Merkel cell polyomavirus (MCPyV)., Results: Sera from patients with distinct autoimmune diseases ( n = 44; mean age 20 years) had a prevalence of MWPyV antibodies of 68%, while in patients with lymphoproliferative disorders ( n = 15; mean age 14 years), subjected to bone marrow transplantation, the prevalence was 47%. In healthy subjects ( n = 66; mean age 13 years), the prevalence of MWPyV antibodies was 67%. Our immunological investigation indicates that MWPyV/HPyV 10 seroconversion occurs early in life and MWPyV/HPyV 10 appears to be another polyomavirus ubiquitous in the human population. A significantly lower MWPyV antibody reactivity together with a lower immunological profile was detected in the sera of LPD patients compared with HS2 (* p < 0.05) (Fisher's exact test). LPD and AD patients have a similar MCPyV seroprevalence compared with healthy subjects., Discussion: MWPyV seroprevalence indicates that this HPyV is not associated with lymphoproliferative and autoimmune diseases. However, the ability to produce high levels of antibodies against MWPyV appears to be impaired in patients with lymphoproliferative disorders. Immunological investigations indicate that MWPyV seroconversion occurs early in life. MCPyV appears to be a ubiquitous polyomavirus, like other HPyVs, in the human population., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Nicol, Mazzoni, Iaquinta, De Pace, Gaboriaud, Maximova, Cason, De Martino, Mazziotta, Coursaget, Touzé, Boz, Comar, Tognon and Martini.)

Mycotoxin contamination of animal feed is a complex issue in both animal wellness and food safety. The most diffused mycotoxins subject to the official control of animal feed are Aflatoxin B1 (AF), Zearalenone (ZEA), Deoxynivalenol (DON), Ochratoxin A (OCRA), Fumonisins (FUMO), and T-2/HT-2 toxins. This work describes the results of five years of monitoring focused on the evaluation of mycotoxin contamination of animal feed. Analytical determinations were carried out by means of accredited ELISA. The obtained results showed a non-alarming scenario, with several samples resulting as "non-compliant" according to the Maximum Residue Limits (MRLs) set in European Regulation No. 574/2011. Out of 722 analyzed samples coming from 2 Italian regions, Apulia and Basilicata, 14 samples were characterized by mycotoxin concentrations higher than related MRL; in particular, 5, 4, and 5 non-compliant samples for DON, AF, and ZEA, respectively. This study also evaluated the possible correlations between mycotoxin type and feed use with a special focus on animal sensitivity to mycotoxins.

Stem cell-based therapy is a significant topic in regenerative medicine, with a predominant role being played by human mesenchymal stem cells (hMSCs). The hMSCs have been shown to be suitable in regenerative medicine for the treatment of bone tissue. In the last few years, the average lifespan of our population has gradually increased. The need of biocompatible materials, which exhibit high performances, such as efficiency in bone regeneration, has been highlighted by aging. Current studies emphasize the benefit of using biomimetic biomaterials, also known as scaffolds, for bone grafts to speed up bone repair at the fracture site. For the healing of injured bone and bone regeneration, regenerative medicine techniques utilizing a combination of these biomaterials, together with cells and bioactive substances, have drawn a great interest. Cell therapy, based on the use of hMSCs, alongside materials for the healing of damaged bone, has obtained promising results. In this work, several aspects of cell biology, tissue engineering, and biomaterials applied to bone healing/regrowth will be considered. In addition, the role of hMSCs in these fields and recent progress in clinical applications are discussed. Impact Statement The restoration of large bone defects is both a challenging clinical issue and a socioeconomic problem on a global scale. Different therapeutic approaches have been proposed for human mesenchymal stem cells (hMSCs), considering their paracrine effect and potential differentiation into osteoblasts. However, different limitations are still to be overcome in using hMSCs as a therapeutic opportunity in bone fracture repair, including hMSC administration methods. To identify a suitable hMSC delivery system, new strategies have been proposed using innovative biomaterials. This review provides an update of the literature on hMSC/scaffold clinical applications for the management of bone fractures.

A novel, air and thermally stable, yet highly reactive trifluoromethylthiolating reagent, CF 3 SO 2 SCF 3 (1), was prepared easily in one step from commercially inexpensive CF 3 SO 2 Na and Tf 2 O. 1 is a highly versatile and atom-efficient reagent that can generate one equivalent of CF 3 S + , two equivalents of CF 3 S - , or a combination of CF 3 S⋅/CF 3 ⋅ species. Many high-yielding CF 3 S reactions of C, O, S, and N-nucleophiles were achieved, including the simple-step preparations of many reported CF 3 S reagents. 1 delivered a hitherto hard-to-synthesize ArOSCF 3 that was followed by a novel CF 3 S II -rearrangement. Through Cu or TDAE/Ph 3 P combinations, 1 generated two equivalents of CF 3 S anion species, and the photo-catalyzed reactions of alkenes with 1 provided CF 3 /CF 3 S-containing products in high atom-efficiency., (© 2023 Wiley-VCH GmbH.)

Anemia is a common complication of chronic kidney disease (CKD) and is associated with increased mortality and reduced health-related quality of life. Anemia is characterized by a decrease in hemoglobin, the iron-rich protein that the body uses for oxygen transport. Iron is required to produce hemoglobin, and disruptions in the iron homeostasis can lead to iron-deficiency anemia. Management of anemia in individuals with CKD is typically performed by a team of physicians, nurse practitioners, physician assistants, or registered nurses. Throughout the care continuum, the management can be enhanced by multidisciplinary care, and individuals with CKD can benefit from the involvement of other specialties, with dietitians/nutritionists playing an important role. However, a key area of unmet clinical need is how to assess and address iron-deficiency anemia. This review aims to provide an overview of iron-deficiency anemia in CKD and how this may be diagnosed and managed by the entire kidney care team, such as describing the mechanisms underlying iron homeostasis, the complications of iron-deficiency anemia, and the current challenges associated with its diagnosis and treatment in CKD. Opportunities for each multidisciplinary team member to add value to the care of individuals with CKD and iron-deficiency anemia are also described., (© 2023 The Authors.)

Spastic paraplegia 50 (SPG50) is an ultrarare childhood-onset neurological disorder caused by biallelic loss-of-function variants in the AP4M1 gene. SPG50 is characterized by progressive spastic paraplegia, global developmental delay, and subsequent intellectual disability, secondary microcephaly, and epilepsy. We preformed preclinical studies evaluating an adeno-associated virus (AAV)/AP4M1 gene therapy for SPG50 and describe in vitro studies that demonstrate transduction of patient-derived fibroblasts with AAV2/AP4M1, resulting in phenotypic rescue. To evaluate efficacy in vivo, Ap4m1-KO mice were intrathecally (i.t.) injected with 5 × 1011, 2.5 × 1011, or 1.25 × 1011 vector genome (vg) doses of AAV9/AP4M1 at P7-P10 or P90. Age- and dose-dependent effects were observed, with early intervention and higher doses achieving the best therapeutic benefits. In parallel, three toxicology studies in WT mice, rats, and nonhuman primates (NHPs) demonstrated that AAV9/AP4M1 had an acceptable safety profile up to a target human dose of 1 × 1015 vg. Of note, similar degrees of minimal-to-mild dorsal root ganglia (DRG) toxicity were observed in both rats and NHPs, supporting the use of rats to monitor DRG toxicity in future i.t. AAV studies. These preclinical results identify an acceptably safe and efficacious dose of i.t.-administered AAV9/AP4M1, supporting an investigational gene transfer clinical trial to treat SPG50.

Cutting up food into small pieces is well known to improve digestion. New work now shows that this concept also applies in the cellular world, by demonstrating that phagolysosome vesiculation promotes cell corpse degradation in Caenorhabditis elegans blastomeres., Competing Interests: Declaration of interests The authors declare no competing Interests., (Published by Elsevier Inc.)

Curcuminoid complex, a mixture of curcumin, demethoxycurcumin and didemethoxycurcumin, is one of the most popular antioxidants of natural origin, and it has a multitude of other health benefits. It is threatened by the proliferation of counterfeit products on the market containing synthetic curcuminoids whose addition is difficult to identify as they present the three curcuminoid forms in the correct ratios. Consequently, the necessity to detect this fraudulent practice is escalating. Carbon-14 analysis is the most effective available method, but it is also expensive and difficult to implement. This paper describes the first attempt to characterize natural curcuminoids and their synthetic form, used as an adulterant, through the analysis of stable isotope ratios of carbon and hydrogen (expressed as δ 13 C and δ 2 H). Carbon values greater than -28.6‱ and hydrogen values greater than -71‱ may indicate the addition of synthetic curcuminoids to the natural ones.

Chediak-Higashi syndrome (CHS) is a rare, autosomal recessive disorder caused by biallelic mutations in the lysosomal trafficking regulator (LYST) gene. Even though enlarged lysosomes and/or lysosome-related organelles (LROs) are the typical cellular hallmarks of CHS, they have not been investigated in human neuronal models. Moreover, how and why the loss of LYST function causes a lysosome phenotype in cells has not been elucidated. We report that the LYST-deficient human neuronal model exhibits lysosome depletion accompanied by hyperelongated tubules extruding from enlarged autolysosomes. These results have also been recapitulated in neurons differentiated from CHS patients' induced pluripotent stem cells (iPSCs), validating our model system. We propose that LYST ensures the correct fission/scission of the autolysosome tubules during autophagic lysosome reformation (ALR), a crucial process to restore the number of free lysosomes after autophagy. We further demonstrate that LYST is recruited to the lysosome membrane, likely to facilitate the fission of autolysosome tubules. Together, our results highlight the key role of LYST in maintaining lysosomal homeostasis following autophagy and suggest that ALR dysregulation is likely associated with the neurodegenerative CHS phenotype., (© 2023. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.)

The growing population in cities is causing a deterioration of air quality due to the emission of pollutants, causing serious health impacts. Trees and urban forests can contribute through the interception and removal of air pollutants such as particulate matter (PM). The dry deposition of PM by vegetation depends on air pollutant concentration, meteorological conditions, and specific leaf traits. Several studies explored the ability of different plant species to accumulate PM on leaf structures leading to the development of models to quantify the PM removal. The i-Tree Eco is the most used model to evaluate ecosystem services provided by urban trees. However, fine particulate matter (PM 2.5 ) removal is still calculated with a poorly evaluated function of deposition velocity (which depends on wind speed and leaf area) without differentiating between tree species. Therefore, we present an improvement of the standard model calculation introducing a leaf trait index to distinguish the species effect on PM net removal. We also compared model results with measurements of deposited leaf PM by vacuum filtration. The index includes the effect of morphological and functional leaf characteristics of tree species using four parameters: leaf water storage, deposition velocity, resuspension rate and leaf washing capacity. Leaves of 11 common urban tree species were sampled in representative areas of the city of Ferrara (Italy) and at different times of the year from 2018 to 2021. This includes four deciduous broadleaf trees (Tilia cordata, Platanus acerifolia, Acer platanoides, Celtis australis), three evergreen broadleaf trees (Quercus ilex, Magnolia grandiflora, Nerium oleander), and four conifers (Thuja orientalis, Cedrus libani, Pinus pinaster, Picea abies). The results provide significant advancement in assessing PM removal using decision support tools such as models to properly select tree species for future urban tree planting programs aimed at improving air quality., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The concept of phytoequivalence was developed in Germany in mid-1990 to indicate that a herbal extract is equivalent to another one, more specifically, to a clinically validated one. The pharmacological/physiological activity of an extract is related to its composition, but, due to the presence of many constituents, reliable technique are necessary to compare extracts. While the concept of equivalence among pure chemicals or isolated substances is feasible using the modern chromatographic and spectroscopic technology, specific guideline for herbal extracts or botanicals are still missing. This is mainly due to the multicomponent nature of the herbal extracts and to the natural variability of their constituents. Nevertheless, by using mathematical and/or chemometric approaches, the concept of phytoequivalence can be precisely addressed, as exemplified by a series of examples that are discussed in detail. Helpful practical recommendations based on literature data and personal experience are presented., (Copyright © 2022. Published by Elsevier B.V.)

Adaptor protein 4 (AP-4) is a heterotetrameric complex composed of ε, β4, μ4, and σ4 subunits that mediates export of a subset of transmembrane cargos, including autophagy protein 9A (ATG9A), from the trans -Golgi network (TGN). AP-4 has received particular attention in recent years because mutations in any of its subunits cause a complicated form of hereditary spastic paraplegia referred to as "AP-4-deficiency syndrome." The identification of proteins that interact with AP-4 has shed light on the mechanisms of AP-4-dependent cargo sorting and distribution within the cell. However, the mechanisms by which the AP-4 complex itself is assembled have remained unknown. Here, we report that the alpha- and gamma-adaptin-binding protein (AAGAB, also known as p34) binds to and stabilizes the AP-4 ε and σ4 subunits, thus promoting complex assembly. The physiological importance of these interactions is underscored by the observation that AAGAB-knockout cells exhibit reduced levels of AP-4 subunits and accumulation of ATG9A at the TGN like those in cells with mutations in AP-4-subunit genes. These findings demonstrate that AP-4 assembly is not spontaneous but AAGAB-assisted, further contributing to the understanding of an adaptor protein complex that is critically involved in development of the central nervous system.

Background: Recent reports of patients with severe, late-stage COVID-19 ARDS with reduced respiratory system compliance described paradoxical decreases in plateau pressure and increases in respiratory system compliance in response to anterior chest wall loading. We aimed to assess the effect of chest wall loading during supine and prone position in ill patients with COVID-19-related ARDS and to investigate the effect of a low or normal baseline respiratory system compliance on the findings., Methods: This is a single-center, prospective, cohort study in the intensive care unit of a COVID-19 referral center. Consecutive mechanically ventilated, critically ill patients with COVID-19-related ARDS were enrolled and classified as higher (≥ 40 ml/cmH 2 O) or lower respiratory system compliance (< 40 ml/cmH 2 O). The study included four steps, each lasting 6 h: Step 1, supine position, Step 2, 10-kg continuous chest wall compression (supine + weight), Step 3, prone position, Step 4, 10-kg continuous chest wall compression (prone + weight). The mechanical properties of the respiratory system, gas exchange and alveolar dead space were measured at the end of each step., Results: Totally, 40 patients were enrolled. In the whole cohort, neither oxygenation nor respiratory system compliance changed between supine and supine + weight; both increased during prone positioning and were unaffected by chest wall loading in the prone position. Alveolar dead space was unchanged during all the steps. In 16 patients with reduced compliance, PaO 2 /FiO 2 significantly increased from supine to supine + weight and further with prone and prone + weight (107 ± 15.4 vs. 120 ± 18.5 vs. 146 ± 27.0 vs. 159 ± 30.4, respectively; p < 0.001); alveolar dead space decreased from both supine and prone position after chest wall loading, and respiratory system compliance significantly increased from supine to supine + weight and from prone to prone + weight (23.9 ± 3.5 vs. 30.9 ± 5.7 and 31.1 ± 5.7 vs. 37.8 ± 8.7 ml/cmH 2 O, p < 0.001). The improvement was higher the lower the baseline compliance., Conclusions: Unlike prone positioning, chest wall loading had no effects on respiratory system compliance, gas exchange or alveolar dead space in an unselected cohort of critically ill patients with C-ARDS. Only patients with a low respiratory system compliance experienced an improvement, with a higher response the lower the baseline compliance., (© 2022. The Author(s).)

One of the hallmarks of Alzheimer's disease is the accumulation of toxic amyloid-β (Aβ) peptides in extracellular plaques. The direct precursor of Aβ is the carboxyl-terminal fragment β (or C99) of the amyloid precursor protein (APP). C99 is detected at elevated levels in Alzheimer's disease brains, and its intracellular accumulation has been linked to early neurotoxicity independently of Aβ. Despite this, the causes of increased C99 levels are poorly understood. Here, we demonstrate that APP interacts with the clathrin vesicle adaptor AP-1 (adaptor protein 1), and we map the interaction sites on both proteins. Using quantitative kinetic trafficking assays, established cell lines and primary neurons, we also show that this interaction is required for the transport of APP from the trans-Golgi network to endosomes. In addition, disrupting AP-1-mediated transport of APP alters APP processing and degradation, ultimately leading to increased C99 production and Aβ release. Our results indicate that AP-1 regulates the subcellular distribution of APP, altering its processing into neurotoxic fragments., Competing Interests: Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

Among European countries, Italy is the most exposed to the risk of biological invasions, principally for its numerous entry points (ports and airports) and for climatic conditions favorable for the acclimatization of several invasive species. Here it was assessed that the greatest threats to our agro-ecosystems come mainly from the passenger baggage in which a variety of fruits and vegetables are carried. From 2016 to 2021, large quantities of plant products were found in the luggage of passengers travelling from outside the EU and seized at the BCPs (border control posts) in the Campania region. Inspections and the following laboratory analyses were conducted on the plant material to assess the presence of exotic pests. Inspections led to several non-native species being recorded, and among the intercepted organisms, some should be considered "alarming", such as Bactrocera dorsalis , Anastrepha obliqua , and Leucinodes africensis . Despite a well-organized border inspection system, travelers transporting infested material unknowingly contribute to increasing the risk of the introduction of exotic species. Given the current situation, it is necessary to impose stricter controls and greater attention, ensuring compliance with the requirements of the new phytosanitary regulations by the actors involved in the transport of plant material. Finally, it is essential to improve awareness through a phytosanitary campaign on plant health risks, especially for people wishing to transport fruits and vegetables in their luggage.

A man in his eighties with acute heart failure and cardiorenal syndrome developed severe hypernatraemia with diuresis. In this situation, palliation is often considered when renal replacement therapy is inappropriate. The literature to guide treatment of dysnatraemia in this setting is limited. Diuretics often worsen hypernatraemia and fluid replacement exacerbates heart failure. We describe a successful approach to this clinical Catch-22: sequential nephron blockade with intravenous 5% dextrose. Seemingly counterintuitive, the natriuretic effect of this combination had not previously been compared with diuretic monotherapy for heart failure. Yet this immediately effective strategy generated a high natriuresis-to-diuresis ratio and functioned as a bridge to cardiac resynchronisation therapy (CRT). In conjunction with a low salt diet, CRT facilitated the maintenance of sodium homeostasis and fluid balance. Thus, by improving the underlying pathophysiology (ie, inadequate cardiac output), CRT may enhance the outcomes of patients with cardiorenal syndrome and hypernatraemia., Competing Interests: Competing interests: Dr RR has received fees for lectures sponsored by Novartis., (© BMJ Publishing Group Limited 2022. No commercial re-use. See rights and permissions. Published by BMJ.)

Purpose: Assess long-term quality of life (HR-QoL) and socio-economic impact in COVID-19-related ARDS (C-ARDS) survivors., Methods: C-ARDS survivors were followed up at 6 months in this prospective, cohort study. HR-QoL was assessed using SF-36 and EQ-5D-5L, and the socio-economic burden of COVID-19 was evaluated with a dedicated questionnaire. Clinical data were prospectively recorded., Results: Seventy-nine survivors, age 63 [57-71], 84% male, were enrolled. The frequency of EQ-5D-5L reported problems was significantly higher among survivors compared to normal, in mobility, usual activities, and self-care; anxiety and depression and pain were not different. SF-36 scores were lower than the reference population, and physical and mental summary scores were below normal in 52% and 33% of the subjects, respectively. In the multivariable analysis, prolonged hospital length of stay (OR 1.45; p 0.02) and two or more comorbidities on admission (OR 7.42; p 0.002) were significant predictors of impaired "physical" and "mental" HR-QoL, respectively. A total of 38% subjects worsened social relations, 42% changed their employment status, and 23% required personal care support., Conclusions: C-ARDS survivors have long-term impairment in HR-QoL and socio-economic problems. Prolonged hospital stay and previous comorbidities are risk factors for developing health-related issues., (© 2022. The Author(s).)

Background: The carotid artery velocity-time integral (CVTI) and the carotid Doppler peak velocity (cDPV), as well as measures of their variation induced by the respiratory cycle, have been proposed as fast and easy to obtain ultrasound measures for assessing fluid responsiveness in intensive care unit patients. To investigate this possibility, we conducted a prospective observational study in hemodynamically unstable patients under mechanical ventilation., Methods: From May 1 to December 31, 2019, we conducted a prospective observational study involving 50 hemodynamically unstable patients under mechanical ventilation. We obtained a total of 800 Doppler ultrasound measurements from the left common carotid artery and at the level of the aortic annulus in the apical five-chamber view. The two measurements were performed before and after a 7 mL/kg fluid challenge and within the first hour of the onset of hemodynamic instability. The maximum Doppler peak velocity, the minimum Doppler peak velocity, and the maximum and minimum VTI at both the aortic and carotid level were acquired., Results: Twenty-eight (56%) patients showed a ≥15% increase in AoVTI after the fluid challenge, and were therefore identified as "fluid responders". All Doppler measurements were always significantly greater (P<0.0001) in fluid responders in relation to both carotid and aortic parameters. Good agreement between the above-mentioned measurements was found: Cohen's kappa coefficient between the carotid and aortic ΔDPV was 0.76 (95% CI 0.58-0.94); and between the Carotid and Aortic ΔVTI it was 0.84 (95% CI 0.68-0.99)., Conclusions: CDPV was found to predict fluid responsiveness in unstable mechanically ventilated patients.

Automated valuation models (AVMs) are widely used by financial institutions to estimate the property value for a residential mortgage. The distribution of pricing errors obtained from AVMs generally show fat tails (Pender 2016; Demiroglu and James Management Science, 64 (4), 1747-1760 2018). The extreme events on the tails are usually known as "black swans" (Taleb 2010) in finance and their existence complicates financial risk management, assessment, and regulation. We show via theory, Monte Carlo experiments, and an empirical example that a direct relation exists between non-normality of the pricing errors and goodness-of-fit of the house pricing models. Specifically, we provide an empirical example using US housing prices where we demonstrate an almost perfect linear relation between the estimated degrees-of-freedom for a Student's t distribution and the goodness-of-fit of sophisticated evaluation models with spatial and spatialtemporal dependence., (© The Author(s) 2021.)

Lysosomes are membrane-bound organelles that degrade diverse biomolecules and regulate a multitude of other essential processes including cell growth and metabolism, signaling, plasma membrane repair and infection. Such diverse functions of lysosomes are highly coordinated in space and time and are therefore tightly coupled to the directional transport of the organelles within the cytoplasm. Thus, robust quantitative assessments of lysosome positioning within the cell provide a valuable tool for researchers interested in understanding these multifunctional organelles. Here, we present point-by-point methodology to measure lysosome positioning by two straight forward and widely used techniques: shell analysis and line scan., (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Niemann-Pick disease, type C (NPC) is a childhood-onset, lethal, neurodegenerative disorder caused by autosomal recessive mutations in the genes NPC1 or NPC2 and characterized by impaired cholesterol homeostasis, a lipid essential for cellular function. Cellular cholesterol levels are tightly regulated, and mutations in either NPC1 or NPC2 lead to deficient transport and accumulation of unesterified cholesterol in the late endosome/lysosome compartment, and progressive neurodegeneration in affected individuals. Previous cell-based studies to understand the NPC cellular pathophysiology and screen for therapeutic agents have mainly used patient fibroblasts. However, these do not allow modeling the neurodegenerative aspect of NPC disease, highlighting the need for an in vitro system that permits understanding the cellular mechanisms underlying neuronal loss and identifying appropriate therapies. This study reports the development of a novel human iPSC-derived, inducible neuronal model of Niemann-Pick disease, type C1 (NPC1)., Results: We generated a null i3Neuron (inducible × integrated × isogenic) (NPC1 -/- i 3 Neuron) iPSC-derived neuron model of NPC1. The NPC1 -/- and the corresponding isogenic NPC1 +/+ i 3 Neuron cell lines were used to efficiently generate homogenous, synchronized neurons that can be used in high-throughput screens. NPC1 -/- i 3 Neurons recapitulate cardinal cellular NPC1 pathological features including perinuclear endolysosomal storage of unesterified cholesterol, accumulation of GM2 and GM3 gangliosides, mitochondrial dysfunction, and impaired axonal lysosomal transport. Cholesterol storage, mitochondrial dysfunction, and axonal trafficking defects can be ameliorated by treatment with 2-hydroxypropyl-β-cyclodextrin, a drug that has shown efficacy in NPC1 preclinical models and in a phase 1/2a trial., Conclusion: Our data demonstrate the utility of this new cell line in high-throughput drug/chemical screens to identify potential therapeutic agents. The NPC1 -/- i 3 Neuron line will also be a valuable tool for the NPC1 research community to explore the pathological mechanisms contributing to neuronal degeneration., (© 2021. The Author(s).)

Trees and urban forests remove particulate matter (PM) from the air through the deposition of particles on the leaf surface, thus helping to improve air quality and reduce respiratory problems in urban areas. Leaf deposited PM, in turn, is either resuspended back into the atmosphere, washed off during rain events or transported to the ground with litterfall. The net amount of PM removed depends on crown and leaf characteristics, air pollution concentration, and weather conditions, such as wind speed and precipitation. Many existing deposition models, such as i-Tree Eco , calculate PM 2.5 removal using a uniform deposition velocity function and resuspension rate for all tree species, which vary based on leaf area and wind speed. However, model results are seldom validated with experimental data. In this study, we compared i-Tree Eco calculations of PM 2.5 deposition with fluxes determined by eddy covariance assessments (canopy scale) and particulate matter accumulated on leaves derived from measurements of vacuum/filtration technique as well as scanning electron microscopy combined with energy-dispersive X-ray spectroscopy (leaf scale). These investigations were carried out at the Capodimonte Royal Forest in Naples. Modeled and measured fluxes showed good overall agreement, demonstrating that net deposition mostly happened in the first part of the day when atmospheric PM concentration is higher, followed by high resuspension rates in the second part of the day, corresponding with increased wind speeds. The sensitivity analysis of the model parameters showed that a better representation of PM deposition fluxes could be achieved with adjusted deposition velocities. It is also likely that the standard assumption of a complete removal of particulate matter, after precipitation events that exceed the water storage capacity of the canopy (Ps), should be reconsidered to better account for specific leaf traits. These results represent the first validation of i-Tree Eco PM removal with experimental data and are a starting point for improving the model parametrization and the estimate of particulate matter removed by urban trees.

Defects in DNA repair frequently lead to neurodevelopmental and neurodegenerative diseases, underscoring the particular importance of DNA repair in long-lived post-mitotic neurons 1,2 . The cellular genome is subjected to a constant barrage of endogenous DNA damage, but surprisingly little is known about the identity of the lesion(s) that accumulate in neurons and whether they accrue throughout the genome or at specific loci. Here we show that post-mitotic neurons accumulate unexpectedly high levels of DNA single-strand breaks (SSBs) at specific sites within the genome. Genome-wide mapping reveals that SSBs are located within enhancers at or near CpG dinucleotides and sites of DNA demethylation. These SSBs are repaired by PARP1 and XRCC1-dependent mechanisms. Notably, deficiencies in XRCC1-dependent short-patch repair increase DNA repair synthesis at neuronal enhancers, whereas defects in long-patch repair reduce synthesis. The high levels of SSB repair in neuronal enhancers are therefore likely to be sustained by both short-patch and long-patch processes. These data provide the first evidence of site- and cell-type-specific SSB repair, revealing unexpected levels of localized and continuous DNA breakage in neurons. In addition, they suggest an explanation for the neurodegenerative phenotypes that occur in patients with defective SSB repair.

Microbiomes of freshwater basins intended for human use remain poorly studied, with very little known about the microbial response to in situ oil spills. Lake Pertusillo is an artificial freshwater reservoir in Basilicata, Italy, and serves as the primary source of drinking water for more than one and a half million people in the region. Notably, it is located in close proximity to one of the largest oil extraction plants in Europe. The lake suffered a major oil spill in 2017, where approximately 400 tons of crude oil spilled into the lake; importantly, the pollution event provided a rare opportunity to study how the lacustrine microbiome responds to petroleum hydrocarbon contamination. Water samples were collected from Lake Pertusillo 10 months prior to and 3 months after the accident. The presence of hydrocarbons was verified and the taxonomic and functional aspects of the lake microbiome were assessed. The analysis revealed specialized successional patterns of lake microbial communities that were potentially capable of degrading complex, recalcitrant hydrocarbons, including aromatic, chloroaromatic, nitroaromatic, and sulfur containing aromatic hydrocarbons. Our findings indicated that changes in the freshwater microbial community were associated with the oil pollution event, where microbial patterns identified in the lacustrine microbiome 3 months after the oil spill were representative of its hydrocarbonoclastic potential and may serve as effective proxies for lacustrine oil pollution.

Background: Vaccine effectiveness relies on various serological tests, whose aim is the measurement of antibody titer in serum samples collected during clinical trials before and after vaccination. Among the serological assays required by the regulatory authorities to grant influenza vaccine release there are: Hemagglutination inhibition (HAI), microneutralization (MN), and Single Radial Hemolysis (SRH). Although antibodies are regarded to be relatively stable, limited evidences on the effect of multiple freeze-thaw cycles on the stability of antibodies in frozen serum samples are available so far. In view of this, the present paper aimed to evaluate the impact of multiple freeze-thaw cycles on influenza antibody stability, performing HAI, MN and SRH assays. Methods: Ten serum samples were divided into 14 aliquots each, stored at -20 °C and taken through a total of 14 freeze-thaw cycles to assess influenza antibody stability. Each assay measurement was carried out following internal procedures based on World Health Organization (WHO) guidelines. Results: No statistically significant effect of 14 freeze-thaw cycles on antibody stability, measured through three different assays, was observed. Conclusions: Collectively, these data demonstrated that specific influenza antibody present in serum samples are stable up to 14 freeze-thaw cycles.

Extremely high temperatures, which negatively affect the human health and plant performances, are becoming more frequent in cities. Urban green infrastructure, particularly trees, can mitigate this issue through cooling due to transpiration, and shading. Temperature regulation by trees depends on feedbacks among the climate, water supply, and plant physiology. However, in contrast to forest or general ecosystem models, most current urban tree models still lack basic processes, such as the consideration of soil water limitation, or have not been evaluated sufficiently. In this study, we present a new model that couples the soil water balance with energy calculations to assess the physiological responses and microclimate effects of a common urban street-tree species (Tilia cordata Mill.) on temperature regulation. We contrast two urban sites in Munich, Germany, with different degree of surface sealing at which microclimate and transpiration had been measured. Simulations indicate that differences in wind speed and soil water supply can be made responsible for the differences in transpiration. Nevertheless, the calculation of the overall energy balance showed that the shading effect, which depends on the leaf area index and canopy cover, contributes the most to the temperature reduction at midday. Finally, we demonstrate that the consideration of soil water availability for stomatal conductance has realistic impacts on the calculation of gaseous pollutant uptake (e.g., ozone). In conclusion, the presented model has demonstrated its ability to quantify two major ecosystem services (temperature mitigation and air pollution removal) consistently in dependence on meteorological and site conditions.

Irrigant solutions commonly used for the treatment of endodontic infections can be inhibited by both organic and inorganic substances. The aim of this study was to evaluate the in vitro antimicrobial activity of the novel irrigant HybenX ® and 2.5% and 5% sodium hypochlorite against Enterococcus faecalis , in presence of dentine powder (DP) or bovine serum albumin 20% (BSA) as inhibitory agents. An E. faecalis American Type Culture Collection (ATCC) 29212 suspension was added to the irrigants (Hybenx ® or NaOCl) and one or two different inhibitors (BSA and DP) either after one-hour pre-incubation at 35 ± 1 °C or not. The antimicrobial activity of HybenX ® against E. faecalis was already proved at 15 min and was neither affected by BSA nor by DP or combinations thereof. NaOCl 2.5% showed an effective antimicrobial activity starting from 15 min and this activity was partially inhibited by BSA and BSA plus DP combination within one hour when pre-incubation occurred. NaOCl 5% showed antimicrobial activity within 15 min, which was inhibited within one hour only in the presence of both BSA and DP regardless of the pre-incubation period. HybenX ® could represent a good alternative to common irrigants for the treatment of E. faecalis endodontic infections, showing a rapid antimicrobial activity not inhibited by organic and inorganic inhibitors.