Trivalent manganese in dissolved forms: Occurrence, speciation, reactivity and environmental geochemical impact.

The cycling processes of elemental manganese (Mn), including the redox reactions of dissolved Mn(III) (dMn(III)), directly and indirectly influences the biogeochemical processes of many elements. Though increasing evidence indicates the widespread presence of dMn(III) mediates the fate of many elements, its role may be currently underestimated. There is both a lack of clear understanding of the historical research framework of dMn(III) and a systematic overview of its geochemical properties and detection methods. Therefore, the primary aim of this review is to outline the understanding of dMn(III) in multiple fields, including soil science, analytical chemistry, biochemistry, geochemistry, and water treatment, and summarize the formation pathways, species forms, and detection methods of dMn(III) in aquatic systems. This review considers how the characteristics of dMn(III), the intermediate formed in the single-electron reaction processes of Mn(II) oxidation and Mn(IV) reduction, determines its participation in environmental geochemical processes. Its widespread presence in diverse water systems and active redox properties coupling with various elements confirm its significant role in natural elemental geochemistry cycle and artificial water treatment processes. Therefore, further investigation into the role of dissolved Mn(III) in aquatic systems is warranted to unravel unexplored coupled elemental redox reaction processes mediated by dissolved Mn(III), filling in the gaps in our understanding of manganese environmental geochemistry, and providing a theoretical basis for recognizing the role of dMn(III) role in water treatment technologies.

Disparate pathways for extrachromosomal DNA biogenesis and genomic DNA repair.

Oncogene amplification on extrachromosomal DNA (ecDNA) is a pervasive driver event in cancer, yet our understanding of how ecDNA forms is limited. Here, we couple a CRISPR-based method for ecDNA induction with extensive characterization of newly formed ecDNA to examine their biogenesis. We find that DNA circularization is efficient, irrespective of 3D genome context, with formation of 800kb, 1 Mb, and 1.8 Mb ecDNAs reaching or exceeding 15%. We show non-homologous end joining and microhomology-mediated end joining both contribute to ecDNA formation, while inhibition of DNA-PKcs and ATM have opposing impacts on ecDNA formation. EcDNA and the corresponding chromosomal excision scar can form at significantly different rates and respond differently to DNA-PKcs and ATM inhibition. Taken together, our results support a model of ecDNA formation in which double strand break ends dissociate from their legitimate ligation partners prior to joining of illegitimate ends to form the ecDNA and excision scar.

Manipulation of photosensory and circadian signaling restricts phenotypic plasticity in response to changing environmental conditions in Arabidopsis.

Plants exploit phenotypic plasticity to adapt their growth and development to prevailing environmental conditions. Interpretation of light and temperature signals is aided by the circadian system, which provides a temporal context. Phenotypic plasticity provides a selective and competitive advantage in nature but is obstructive during large-scale, intensive agricultural practices since economically important traits (including vegetative growth and flowering time) can vary widely depending on local environmental conditions. This prevents accurate prediction of harvesting times and produces a variable crop. In this study, we sought to restrict phenotypic plasticity and circadian regulation by manipulating signaling systems that govern plants' responses to environmental signals. Mathematical modeling of plant growth and development predicted reduced plant responses to changing environments when circadian and light signaling pathways were manipulated. We tested this prediction by utilizing a constitutively active allele of the plant photoreceptor phytochrome B, along with disruption of the circadian system via mutation of EARLY FLOWERING3. We found that these manipulations produced plants that are less responsive to light and temperature cues and thus fail to anticipate dawn. These engineered plants have uniform vegetative growth and flowering time, demonstrating how phenotypic plasticity can be limited while maintaining plant productivity. This has significant implications for future agriculture in both open fields and controlled environments.

Quantification of First Metatarsal Joint Surface Interactions in Hallux Rigidus Using Distance and Coverage Mapping: A Case-Control Study.

BACKGROUND: Weightbearing cone-beam computed tomography (WBCT) has proven useful for analysis of structural changes of the foot and ankle when compared to conventional radiographs. WBCT allows for extraction of distance and coverage mapping metrics, which may provide novel insight into hallux rigidus (HR). This study retrospectively assessed HR joint space using distance and coverage mapping in a case-control study. METHODS: WBCT images of the foot and ankle for 20 symptomatic HR and 20 control patients were obtained. Three-dimensional models were created and analyzed using a custom semiautomatic measurement algorithm. Distance and coverage mapping metrics for the first metatarsophalangeal and metatarsosesamoid joints were extracted from the models and compared between cohorts. Relationships between these metrics and visual analog scale (VAS) scores, a patient-reported outcome of pain, were assessed in HR patients. RESULTS: Overall first metatarsophalangeal joint space narrowing was noted in HR patients when compared to controls by an average of 11.8% (P = .02). However, no significant changes in the overall coverage of the joint were noted. Decreased joint space width and increased surface-to-surface coverage were only and particularly observed at the plantar medial quadrant of the first metatarsal head in HR patients relative to controls. VAS score was significantly but weakly correlated with dorsolateral quadrant coverage (R2 = 0.26, P = .03). CONCLUSION: Distance and coverage mapping serve as a complementary option to current techniques of quantifying HR changes. These metrics can expand the scope of future work investigating joint articulation changes in HR.

Demographic characteristics, gambling engagement, mental health, and associations with harmful gambling risk among UK Armed Forces serving personnel.

INTRODUCTION: Harmful gambling negatively impacts individuals, families and communities. Growing international evidence indicates that the Armed Forces (AF) community may be at a comparatively higher risk of experiencing harm from gambling than the general population. The current study sought to identify general predictors of harmful gambling and gambling engagement among UK AF serving personnel (AFSP). METHODS: We conducted a cross-sectional, exploratory survey to identify associations between demographic factors, mental health, gambling engagement and gambling type in a sample (N=608) of AFSP. RESULTS: Most of the sample reported past-year gambling, with 23% having experienced harm. Male gender, younger age and lower educational attainment all predicted harmful gambling, as did mental health variables of prior generalised anxiety and post-traumatic stress symptomatology. Strategy-based gambling and online sports betting were also predictive of experiencing harm from gambling. CONCLUSIONS: The risk of harm from gambling is associated with demographic, mental health and gambling engagement variables among AFSP. Better understanding of these predictors is important for the development of individualised treatment approaches for harmful gambling.

A Novel Use of Embryonic Gut Organoid Culture to Investigate Duodenal Atresia.

BACKGROUND: The cause of duodenal atresia (DA) is not known. Tandler's "solid cord" hypothesis conflicts with current biological evidence. In humans, a genetic aetiology is supported by the association with Trisomy 21. Interruption of Fgf10 is the strongest genetic link to DA in mice, demonstrating an increased incidence and severity as embryos mature. This project aimed to develop an organoid model to facilitate ex vivo DA research on the FGF10/FGFR2b signalling pathway. We hypothesised that DA morphology represents an evolving spectrum of disease and that Fgf10 knockout organoids would vary in growth pattern compared to wild-type. METHODS: Organoids were cultured from the duodenum of E12.5 Fgf10 knockout, heterozygous and wild-type embryos, using an air-liquid interface with Growth Factor reduced Matrigel. Organoids were photographed every 48 h to observe growth. Organoids were isolated and fixed after 14 days, then stained with DAPI, KI-67, and cytokeratin to demonstrate proliferation and differentiation. RESULTS: Wild-type duodenum developed into crypt-forming organoids. Fgf10 heterozygous duodenum failed to progress beyond the development stage of spheroids. Fgf10 knockout duodenum failed to demonstrate any growth. Wholemount staining showed the greatest cell proliferation and differentiation in wild-type tissue. CONCLUSION: This research presents a novel concept for the growth of embryonic gastrointestinal tissue to inform normal biology. The small sample numbers and restricted culture duration limit longer-term growth analysis. While this model serves as a potential ex vivo setting for future research, that research should consider organoid models with greater standardisation and other gastrointestinal regions. LEVEL OF EVIDENCE: Animal/laboratory study.

BeGraft Aortic Stents: A European Multi-Centre Experience Reporting Acute Safety and Efficacy Outcomes for the Treatment of Vessel Stenosis in Congenital Heart Diseases.

BACKGROUND: Stent implantation has become the preferred method of treatment for treating vessel stenosis in congenital heart diseases. The availability of covered stents may decrease complications and have an important role in the management of patients with complex anatomy. AIM: This study aims to evaluate the feasibility and safety of the pre-mounted cobalt-chromium stent-graft-covered ePTFE Aortic BeGraft in a broad spectrum of vascular lesions. METHODS: This is a multicenter retrospective results analysis of 107 implanted BeGraft stents between 2016 and 2022 in six different European centers. RESULTS: One hundred and four patients with a mean age of thirteen years (range 1-70 years) and with the body weight of 56.5 kg (range 11-115 kg) underwent the BeGraft stent implantation. Stents were implanted in the following conditions: aortic coarctation (74 patients), RVOT dysfunction (12 patients), Fontan circulation (7 patients), and miscellaneous (11 subjects with complex CHD). All the stents were implanted successfully. The median stent diameter was 16 mm (range 7-24 mm), and the median length was 39 mm (range 19-49 mm). Major complications occurred in five subjects (4.7%). During a median follow-up of fourteen (1-70) months, stents' re-dilatation was performed in five patients. CONCLUSIONS: The BeGraft stent can be used safely and effectively in a wide spectrum of congenital heart diseases. Whether these good results will be stable in the longer term still needs to be investigated in a follow-up given its recent introduction into clinical practice, in particular regarding stent fracture or neointimal proliferation.

Contributing Factors to Endothelial Dysfunction in Individuals with Spinal Cord Injuries.

Background: Patients with spinal cord injuries (SCIs) are at a greater risk for the development of cardiovascular diseases (CVDs) than able-bodied individuals due to the high risk of endothelial dysfunction. Summary: For instance, patients with SCIs lose autonomic control of the heart and vasculature, which results in severe fluctuations in blood pressure. These oscillations between hypotension and hypertension have been shown to damage blood vessel endothelial cells and may contribute to the development of atherosclerosis. Furthermore, the loss of skeletal muscle control results in skeletal muscle atrophy and inward remodeling of the conduit arteries. It has been shown that blood vessels in the legs are chronically exposed to high shear, while the aorta experiences chronically low shear. These alterations to shear forces may adversely impact endothelial vasodilatory capacity and promote inflammatory signaling and leukocyte adherence. Additionally, microvascular endothelial vasodilatory capacity is impaired in patients with an SCI, and this may precede changes in conduit artery endothelial function. Finally, due to immobility and a loss of skeletal muscle mass, patients with SCIs have a higher risk of metabolic disorders, inflammation, and oxidative stress. Key Messages: Collectively, these factors may impair endothelium-dependent vasodilatory capacity, promote leukocyte adhesion and infiltration, promote the peroxidation of lipids, and ultimately support the development of atherosclerosis. Therefore, future interventions to prevent CVDs in patients with SCIs should focus on the management of endothelial health to prevent endothelial dysfunction and atherosclerosis.

The effect of repeat feeding on dengue virus transmission potential in Wolbachia-infected Aedes aegypti following extended egg quiescence.

As Wolbachia pipientis is more widely being released into field populations of Aedes aegypti for disease control, the ability to select the appropriate strain for differing environments is increasingly important. A previous study revealed that longer-term quiescence in the egg phase reduced the fertility of mosquitoes, especially those harboring the wAlbB Wolbachia strain. This infertility was also associated with a greater biting rate. Here, we attempt to quantify the effect of this heightened biting behavior on the transmission potential of the dengue virus using a combination of assays for fitness, probing behavior, and vector competence, allowing repeat feeding, and incorporate these effects in a model of R0. We show that Wolbachia-infected infertile mosquitoes are more interested in feeding almost immediately after an initial blood meal relative to wild type and Wolbachia-infected fertile mosquitoes and that these differences continue for up to 8 days over the period we measured. As a result, the infertile Wolbachia mosquitoes have higher virus prevalence and loads than Wolbachia-fertile mosquitoes. We saw limited evidence of Wolbachia-mediated blocking in the disseminated tissue (legs) in terms of prevalence but did see reduced viral loads. Using a previously published estimate of the extrinsic incubation period, we demonstrate that the effect of repeat feeding/infertility is insufficient to overcome the effects of Wolbachia-mediated blocking on R0. These estimates are very conservative, however, and we posit that future studies should empirically measure EIP under a repeat feeding model. Our findings echo previous work where periods of extensive egg quiescence affected the reproductive success of Wolbachia-infected Ae. aegypti. Additionally, we show that increased biting behavior in association with this infertility in Wolbachia-infected mosquitoes may drive greater vector competence. These relationships require further exploration, given their ability to affect the success of field releases of Wolbachia for human disease reduction in drier climates where longer egg quiescence periods are expected.

Navigating complexities: clinicians' experiences and systemic challenges in the implementation of evidence-based practice for chronic low back pain - a qualitative study.

PURPOSE: Implementation of evidence-based practice (EBP) for chronic low back pain (CLBP) is poor. This study aimed to investigate the factors that influence exercise-based clinicians' implementation of EBP for people with CLBP. MATERIALS AND METHODS: Semi-structured interviews were conducted with 40 clinicians (20 physiotherapists and 20 accredited exercise physiologists). Interviews explored clinicians' definition of EBP, understanding of EBP in CLBP management, barriers and enablers to EBP, and the impact of these factors on EBP implementation. Interviews were analysed using reflexive thematic analysis. RESULTS: A complex interplay among clinician factors, systemic factors, and patient interactions was identified. Clinicians encountered challenges stemming from their misconceptions about EBP, which influenced their perspective(s) on its implementation. Clinicians expressed the challenges of implementing EBP when navigating patient expectations and beliefs, experiencing external pressures from funders and business models, and the subsequent impacts on their well-being. Continuing professional development, support from community, and workplaces that prioritised EBP enhanced clinicians' ability to make decisions that accommodate for the complexities of CLBP and better align with EBP. CONCLUSION: A variety of factors impact exercise-based clinicians' implementation of EBP. The interaction between these factors greatly influences clinicians' ability to engage in and implement EBP in clinical practice.

The interplay among clinician, patient, and health system/structural level factors affects exercise-based clinicians' implementation of evidence-based practice for chronic low back pain.Clinicians are well positioned to identify issues in clinical practice and initiate appropriate changes that could improve evidence-based practice.Clinicians could provide feedback on continuing professional development to ensure it is accessible and applicable to clinical practice.Clinicians could encourage workplaces to prioritise time and/or funding for sufficient engagement in evidence-based practice.

The Effect of Creatine Supplementation on Resistance Training-Based Changes to Body Composition: A Systematic Review and Meta-analysis.

ABSTRACT: Desai, I, Wewege, MA, Jones, MD, Clifford, BK, Pandit, A, Kaakoush, NO, Simar, D, and Hagstrom, AD. The effect of creatine supplementation on resistance training-based changes to body composition: A systematic review and meta-analysis. J Strength Cond Res XX(X): 000-000, 2024-The purpose of this review was to determine the added effect of creatine supplementation on changes in body composition with resistance training in adults younger than 50 years. The review protocol was preregistered on the Open Science Framework (osf.io/x48a6/). Our primary outcome was lean body mass (LBM); secondary outcomes were body fat percentage (%) and body fat mass (kg). We performed a random-effects meta-analysis in R using the metafor package. Subgroup analyses were conducted to examine the effects of training status and use of a carbohydrate drink with creatine. We conducted a meta-regression to examine the moderating effect of total training volume. Statistical significance was set at p < 0.05. One thousand six hundred ninety-four records were screened, and 67 full-text articles were assessed for eligibility. Twelve studies were included in the meta-analysis. Fifty-two percentages of the studies had low risk, 41% some concerns, and 7% high risk of bias. Compared with resistance training (RT) alone, creatine supplementation increased LBM by 1.14 kg (95% CI 0.69 to 1.59), and reduced body fat percentage by -0.88% (95% CI -1.66 to -0.11) and body fat mass by -0.73 kg (95% CI -1.34 to -0.11). There were no differences between training status or carbohydrate subgroups. Training volume was not associated with effect size in all outcomes; 7 g or 0.3 g/kg of body mass of creatine per day is likely to increase LBM by 1 kg and reduce fat mass by 0.7 kg more than RT alone. Concurrent carbohydrate ingestion did not enhance the hypertrophy benefits of creatine.

CoRAL accurately resolves extrachromosomal DNA genome structures with long-read sequencing.

Extrachromosomal DNA (ecDNA) is a central mechanism for focal oncogene amplification in cancer, occurring in approximately 15% of early-stage cancers and 30% of late-stage cancers. EcDNAs drive tumor formation, evolution, and drug resistance by dynamically modulating oncogene copy-number and rewiring gene-regulatory networks. Elucidating the genomic architecture of ecDNA amplifications is critical for understanding tumor pathology and developing more effective therapies. Paired-end short-read (Illumina) sequencing and mapping have been utilized to represent ecDNA amplifications using a breakpoint graph, where the inferred architecture of ecDNA is encoded as a cycle in the graph. Traversals of breakpoint graph have been used to successfully predict ecDNA presence in cancer samples. However, short-read technologies are intrinsically limited in the identification of breakpoints, phasing together of complex rearrangements and internal duplications, and deconvolution of cell-to-cell heterogeneity of ecDNA structures. Long-read technologies, such as from Oxford Nanopore Technologies, have the potential to improve inference as the longer reads are better at mapping structural variants and are more likely to span rearranged or duplicated regions. Here, we propose CoRAL (Complete Reconstruction of Amplifications with Long reads), for reconstructing ecDNA architectures using long-read data. CoRAL reconstructs likely cyclic architectures using quadratic programming that simultaneously optimizes parsimony of reconstruction, explained copy number, and consistency of long-read mapping. CoRAL substantially improves reconstructions in extensive simulations and 10 datasets from previously-characterized cell lines as compared to previous short and long-read based tools. As long-read usage becomes wide-spread, we anticipate that CoRAL will be a valuable tool for profiling the landscape and evolution of focal amplifications in tumors.

Simple autonomous agents can enhance creative semantic discovery by human groups.

Innovation is challenging, and theory and experiments indicate that groups may be better able to identify and preserve innovations than individuals. But innovation within groups faces its own challenges, including groupthink and truncated diffusion. We performed experiments involving a game in which people search for ideas in various conditions: alone, in networked social groups, or in networked groups featuring autonomous agents (bots). The objective was to search a semantic space of 20,000 nouns with defined similarities for an arbitrary noun with the highest point value. Participants (N = 1875) were embedded in networks (n = 125) of 15 nodes to which we sometimes added 2 bots. The bots had 3 possible strategies: they shared a random noun generated by their immediate neighbors, or a noun most similar from among those identified, or a noun least similar. We first confirm that groups are better able to explore a semantic space than isolated individuals. Then we show that when bots that share the most similar noun operate in groups facing a semantic space that is relatively easy to navigate, group performance is superior. Simple autonomous agents with interpretable behavior can affect the capacity for creative discovery of human groups.

Linking haploinsufficiency of the autism- and schizophrenia-associated gene Cyfip1 with striatal-limbic-cortical network dysfunction and cognitive inflexibility.

Impaired behavioural flexibility is a core feature of neuropsychiatric disorders and is associated with underlying dysfunction of fronto-striatal circuitry. Reduced dosage of Cyfip1 is a risk factor for neuropsychiatric disorder, as evidenced by its involvement in the 15q11.2 (BP1-BP2) copy number variant: deletion carriers are haploinsufficient for CYFIP1 and exhibit a two- to four-fold increased risk of schizophrenia, autism and/or intellectual disability. Here, we model the contributions of Cyfip1 to behavioural flexibility and related fronto-striatal neural network function using a recently developed haploinsufficient, heterozygous knockout rat line. Using multi-site local field potential (LFP) recordings during resting state, we show that Cyfip1 heterozygous rats (Cyfip1+/-) harbor disrupted network activity spanning medial prefrontal cortex, hippocampal CA1 and ventral striatum. In particular, Cyfip1+/- rats showed reduced influence of nucleus accumbens and increased dominance of prefrontal and hippocampal inputs, compared to wildtype controls. Adult Cyfip1+/- rats were able to learn a single cue-response association, yet unable to learn a conditional discrimination task that engages fronto-striatal interactions during flexible pairing of different levers and cue combinations. Together, these results implicate Cyfip1 in development or maintenance of cortico-limbic-striatal network integrity, further supporting the hypothesis that alterations in this circuitry contribute to behavioural inflexibility observed in neuropsychiatric diseases including schizophrenia and autism.

Selective Macrocyclization of Unprotected Peptides with an Ex Situ Gaseous Linchpin Reagent.

Peptide cyclization has dramatic effects on a variety of important properties, enhancing metabolic stability, limiting conformational flexibility, and altering cellular entry and intracellular localization. The hydrophilic, polyfunctional nature of peptides creates chemoselectivity challenges in macrocyclization, especially for natural sequences without biorthogonal handles. Herein, we describe a gaseous sulfonyl chloride derived reagent that achieves amine-amine, amine-phenol, and amine-aniline crosslinking through a minimalist linchpin strategy that affords macrocyclic urea or carbamate products. The cyclization reaction is metal-mediated and involves a novel application of sulfine species that remains unexplored in aqueous or biological contexts. The aqueous method delivers unique cyclic or bicyclic topologies directly from a variety of natural bioactive peptides without the need for protecting-group strategies.

Templated Synthesis of Copper Nanoclusters with a Hybrid Lysozyme-Polymer Material for Enhanced Fluorescence.

Hybrid materials that combine organic polymers and biomacromolecules offer unique opportunities for precisely controlling 3D chemical environments. Although biological or organic templates have been separately used to control the growth of inorganic nanoclusters, hybrid structures represent a relatively unexplored approach to tailoring nanocluster properties. Here, we demonstrate that a molecularly defined lysozyme-polymer resin material acts as a structural scaffold for the synthesis of copper nanoclusters (CuNCs) with well controlled size distributions. The resulting CuNCs have significantly enhanced fluorescence compared with syntheses based on polymeric or biological templates alone. The synergistic approach described here is appealing for the synthesis of biocompatible fluorescent labels with improved photostability.

Electronic-Screening, Brief Intervention and Referral to Treatment (e-SBIRT) for Addictive Disorders: Systematic Review and Meta-Analysis.

BACKGROUND: Addictive disorders are significant global public health burdens. Treatment uptake with these disorders is low and outcomes can be mixed. Electronic screening, brief intervention, and referral to treatment (e-SBIRT) programs have potential to improve uptake and treatment outcomes. To date, however, no prior review of the literature has been conducted to gauge the effectiveness of e-SBIRT for addictive disorders. METHODS: We conducted a systematic review and meta-analysis of the literature concerning e-SBIRT for addictive disorders by surveying the MEDLINE, PubMed, Web of Science, Scopus, Embase, and PsycInfo databases on January 17, 2023. RESULTS: Ten articles were included at analysis reporting evaluation of e-SBIRT interventions for substance use disorders including alcohol use in a variety of settings. No articles were identified regarding treatment for behavioral addictions such as disordered/harmful gambling. Meta-analysis found e-SBIRT to be effective at reducing drinking frequency in the short term only. e-SBIRT was not found to be advantageous over control conditions for abstinence or other treatment outcomes. We identified and described common components of e-SBIRT programs and assessed the quality of available evidence, which was generally poor. CONCLUSION: The present findings suggest that research regarding e-SBIRT is concentrated exclusively on higher-risk substance use. There is a lack of consensus regarding the effectiveness of e-SBIRT for addictive disorders. Although common features exist, e-SBIRT designs are variable, which complicates identification of the most effective components. Overall, the quality of outcome evidence is low, and furthermore, high-quality experimental treatment evaluation research is needed.

B cells in the pneumococcus-infected lung are heterogeneous and require CD4+ T cell help including CD40L to become resident memory B cells.

Recovery from respiratory pneumococcal infections generates lung-localized protection against heterotypic bacteria, mediated by resident memory lymphocytes. Optimal protection in mice requires re-exposure to pneumococcus within days of initial infection. Serial surface marker phenotyping of B cell populations in a model of pneumococcal heterotypic immunity revealed that bacterial re-exposure stimulates the immediate accumulation of dynamic and heterogeneous populations of B cells in the lung, and is essential for the establishment of lung resident memory B (BRM) cells. The B cells in the early wave were activated, proliferating locally, and associated with both CD4+ T cells and CXCL13. Antagonist- and antibody-mediated interventions were implemented during this early timeframe to demonstrate that lymphocyte recirculation, CD4+ cells, and CD40 ligand (CD40L) signaling were all needed for lung BRM cell establishment, whereas CXCL13 signaling was not. While most prominent as aggregates in the loose connective tissue of bronchovascular bundles, morphometry and live lung imaging analyses showed that lung BRM cells were equally numerous as single cells dispersed throughout the alveolar septae. We propose that CD40L signaling from antigen-stimulated CD4+ T cells in the infected lung is critical to establishment of local BRM cells, which subsequently protect the airways and parenchyma against future potential infections.