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Hematopoietic stem cell (HSC)-independent lymphopoiesis has been elucidated in murine embryos. However, our understanding regarding human embryonic counterparts remains limited. Here, we demonstrated the presence of human yolk sac-derived lymphoid-biased progenitors (YSLPs) expressing CD34 , IL7R , LTB , and IRF8 at Carnegie stage 10, much earlier than the first HSC emergence. The number and lymphopoietic potential of these progenitors were both significantly higher in the yolk sac than the embryo proper at this early stage. Importantly, single-cell/bulk culture and CITE-seq have elucidated the tendency of YSLP to differentiate into innate lymphoid cells and dendritic cells. Notably, lymphoid progenitors in fetal liver before and after HSC seeding displayed distinct transcriptional features, with the former closely resembling those of YSLPs. Overall, our data identified the origin, potential, and migratory dynamics of innate lymphoid-biased multipotent progenitors in human yolk sac before HSC emergence, providing insights for understanding the stepwise establishment of innate immune system in humans. [Display omitted] • Progenitors with non-NK lymphoid potential exist in human yolk sac before HSC onset • Human yolk sac lymphoid progenitors before HSC emergence show ILC and DC potential • ILC potential before HSC onset is stronger in human yolk sac than in embryo proper • Early fetal liver lymphoid progenitors share characteristics with those from yolk sac Liu et al. demonstrated the presence of yolk sac-derived lymphoid progenitors expressing IL7R and IRF8 well before HSC emergence in human embryos, which showed differentiation bias toward innate lymphoid cells and dendritic cells. They shared transcriptional profiles with lymphoid progenitors in the fetal liver before HSC colonization. [ABSTRACT FROM AUTHOR]

The mechanisms leading to a predominantly hypertrophied phenotype versus a predominantly oxidative phenotype, the hallmarks of resistance training (RT) or aerobic training (AT), respectively, are being unraveled. In humans, exposure of naïve persons to either AT or RT results in their skeletal muscle exhibiting generic 'exercise stress-related' signaling, transcription, and translation responses. However, with increasing engagement in AT or RT, the responses become refined, and the phenotype typically associated with each form of exercise emerges. Here, we review some of the mechanisms underpinning the adaptations of how muscles become, through AT, 'fit' and RT, 'mighty.' Much of our understanding of molecular exercise physiology has arisen from targeted analysis of post-translational modifications and measures of protein synthesis. Phosphorylation of specific residue sites has been a dominant focus, with canonical signaling pathways (AMPK and mTOR) studied extensively in the context of AT and RT, respectively. These alone, along with protein synthesis, have only begun to elucidate key differences in AT and RT signaling. Still, key yet uncharacterized differences exist in signaling and regulation of protein synthesis that drive unique adaptation to AT and RT. Omic studies are required to better understand the divergent relationship between exercise and phenotypic outcomes of training. [Display omitted] • Skeletal muscle phenotype is underpinned by constituent proteins' activity/function. • Targeted analyses demarcate few differences between exercise modes. • Unknown unique signaling between aerobic/resistance exercise underpin phenotypes. • Multi-timepoint and unbiased omic methods will characterize exercise adaptation. [ABSTRACT FROM AUTHOR]

Based on hydrodynamic theory, blocking the dentinal tubules can reduce discomfort caused by dentin hypersensitivity. This study identified the crystals formed in dentinal tubules from tricalcium silicate (TCS) in phosphate-buffered saline (PBS) and evaluated the effect of PBS concentration on crystal formation. Sixty-nine specimens were made by isolating the cervical part of extracted premolars. TCS was applied by brushing for 10,000 strokes on dentin surface simulating sensitive dentin. Specimens were stored in PBS or solutions with concentrations 1/100, 1/10, 10, and 100 times that of PBS for 1, 30, 60, or 90 days (n = 3). Another nine specimens applied TCS, were immersed in PBS for 3 months, and divided into three subgroups: no treatment, sonication for 10 min, and 1M acetic acid treatment for 3 min. Crystal formation was examined using a scanning electron microscope, assigned five grade scores (0–4) according to maturation, and analyzed by a nonparametric two-way ANOVA (α = 0.05). Crystal components were analyzed using X-ray diffraction (XRD) and energy dispersion X-ray spectroscopy (EDS). The maturation of intratubular crystals was dependent on time and PBS concentration (P < 0.05). In all periods, the high-concentration group showed a higher maturation grade than the low-concentration group. Intratubular crystals were similar to hydroxyapatite according to XRD and EDS, and they withstood sonication and acid application. TCS with nanosized particles formed hydroxyapatite-like crystals in the dentinal tubules, which were dependent on time and concentration of PBS and withstood sonication and acid application. [ABSTRACT FROM AUTHOR]

We have previously demonstrated that dietary saturated fatty acids (SFA), when compared to polyunsaturated fatty acids (PUFA), are preferentially partitioned into oxidation pathways. However, it remains unclear if this preferential handling is maintained when hepatocellular metabolism is shifted toward fatty acid (FA) esterification and away from oxidation, such as when hepatic de novo lipogenesis (DNL) is upregulated. To investigate whether an acute upregulation of hepatic DNL influences dietary FA partitioning into oxidation pathways. 20 healthy volunteers (11 females) underwent a fasting baseline visit followed by two study days, 2-weeks apart. Prior to each study day, participants consumed an isocaloric high-carbohydrate diet (to upregulate hepatic DNL) for 3-days. On the two study days, participants consumed an identical standardised test meal that contained either [U13C]palmitate or [U13C]linoleate, in random order, to trace the fate of dietary FA. Blood and breath samples were collected over a 6h postprandial period and 13C enrichment in breath CO 2 and plasma lipid fractions were measured using gas-chromatography-combustion-isotope ratio mass spectrometry. Compared to the baseline visit, fasting plasma triglyceride concentrations and markers of hepatic DNL, the lipogenic and stearyl-CoA desaturase indices, were significantly (p < 0.05) increased after consumption of the high-carbohydrate diet. Appearance of 13C in expired CO 2 and tracer recovery were significantly (p < 0.05) higher after consumption of the meal containing [U13C]linoleate compared to [U13C]palmitate (5.1 ± 0.5% vs. 3.7 ± 0.4%), respectively. Incorporation of 13C into the plasma triglyceride and non-esterified fatty acid pool was significantly (p < 0.001) greater for [U13C]palmitate compared to [U13C]linoleate. Dietary PUFA compared to SFA appear to be preferentially partitioned into oxidation pathways during an acute upregulation of hepatic DNL, thus consumption of a PUFA-enriched diet may help mitigate intrahepatic triglyceride accumulation in individuals at risk of cardiometabolic disease. [ABSTRACT FROM AUTHOR]

Peripheral nerve injuries (PNIs) are common, with complex defects posing a significant reconstructive challenge. Although vascularised (VNGs) and non-vascularised nerve grafts (NVNGs) are established treatment options, there is no comprehensive summary of the evidence supporting their clinical, electrophysiological, and histological outcomes. This review aims to systematically evaluate the clinical and laboratory literature comparing VNGs and NVNGs to inform future clinical practice and research. This review was prospectively registered and reported according to PRISMA guidelines. PubMed, EMBASE, SCOPUS, and the Cochrane Register were systematically searched. Studies comparing VNGs and NVNGs in PNIs were included. Meta-analyses were performed for outcomes reported in ≥3 laboratory studies. Functional outcomes were synthesised by vote-counting based on direction of effect for clinical studies. Risk-of-bias was assessed using RoB2, ROBINS-I, and SYRCLE, and the certainty of evidence was evaluated using GRADE. Seven clinical and 34 laboratory studies were included. Of the clinical comparisons, 90% and 56% identified an effect on recovery of sensibility (p = 0.01) and motor function (p = 0.05), respectively, that favoured VNGs. Nine (of 13) separate meta-analyses of laboratory studies demonstrated reduced muscular atrophy, superior axonal regeneration, and remyelination in VNGs. VNGs eliminated the 3-day interval of ischaemia otherwise sustained by NVNGs. Overall, the quality of evidence was low. This systematic review indicates that VNGs may offer some advantages over NVNGs in PNI reconstruction. However, due to the low quality of evidence, significant statistical heterogeneity, and clinical diversity of the included studies, these conclusions should be interpreted with caution. Further high-quality clinical trials are necessary to validate these findings. [ABSTRACT FROM AUTHOR]

The hypothalamus regulates homeostasis across the lifespan and is emerging as a regulator of aging. In murine models, aging-related changes in the hypothalamus, including microinflammation and gliosis, promote accelerated neurocognitive decline. We investigated relationships between hypothalamic microstructure and features of neurocognitive aging, including cortical thickness and cognition, in a cohort of community-dwelling older adults (age range 65–97 years, n=124). Hypothalamic microstructure was evaluated with two magnetic resonance imaging diffusion metrics: mean diffusivity (MD) and fractional anisotropy (FA), using a novel image processing pipeline. Hypothalamic MD was cross-sectionally positively associated with age and it was negatively associated with cortical thickness. Hypothalamic FA, independent of cortical thickness, was cross-sectionally positively associated with neurocognitive scores. An exploratory analysis of longitudinal neurocognitive performance suggested that lower hypothalamic FA may predict cognitive decline. No associations between hypothalamic MD, age, and cortical thickness were identified in a younger control cohort (age range 18–63 years, n=99). To our knowledge, this is the first study to demonstrate that hypothalamic microstructure is associated with features of neurocognitive aging in humans. • In animal models, aging-related changes in the hypothalamus drive brain aging. • The role of the hypothalamus in neurocognitive aging in humans is unknown. • We show that hypothalamic microstructure is altered with aging in older adults. • Hypothalamic microstructure is associated with cortical thickness and cognition. [ABSTRACT FROM AUTHOR]

Sarcopenia is a common and natural condition in the elderly, and leads to loss of muscle mass and function. In the presbylarynx there is atrophy of the vocal folds, however the degree of vocal muscle atrophy is poorly studied and the results are contradictory. Aims: The aim of this study was to evaluate the effect of aging on the vocal muscle (thyroarytenoid muscle). Thirteen larynxes removed during necropsy from 13 cadavers were included into two age groups: Control group – G1 (n5), between 25 and 40 years; Elderly Group - GI (n-8), aged 70 years or older. The vocal folds were dissected and prepared for scanning electron microscopy, ensuring a cross-section in the musculature area to allow measurements of muscle fiber parameters. Images were analyzed and photographed at different magnifications. Through the ImageJ software, ten distinct fields of each part were selected. Parameters studied: area, perimeters and diagonals of the thyroarytenoid muscle bundles of both groups. The cross-sectional areas, perimeters and diagonals of the muscle fibers of the thyroarytenoid muscle of the elderly group were significantly smaller than those of the control group. We demonstrated that the vocal muscle is affected in the presbylarynx, with a reduction of its muscle fibers, corresponding to muscle atrophy. However, these findings may not be directly related to vocal symptoms because the patient may develop muscle compensatory mechanisms capable of reducing glottic insufficiency [ABSTRACT FROM AUTHOR]

Innate lymphoid cells (ILCs) are a group of lymphocytes that are devoid of antigen-specific receptors and are mainly found in tissues. The subtypes ILC1, 2, and 3 mirror T-cell functionality in terms of cytokine production and expression of key transcription factors. Although the majority of ILCs are found in tissue (tILCs), they have also been described within the circulation (cILCs). As a result of their better accessibility and putative prognostic value, human cILCs are getting more and more attention in clinical research. However, cILCs are in many aspects functionally distinct from their tILC counterparts. In fact, from the 3 ILC subsets found within the circulation, only for cILC2s could a clear functional correspondence to their tissue counterparts be established. Indeed, cILC2s are emerging as a major driver of allergic reactions with a particular role in asthma. In contrast, recent studies revealed that cILC1s and cILC3s are predominantly in an immature state and constitute progenitors for natural killer cells and ILCs, respectively. We provide an overview about the phenotype and function of the different cILC subtypes compared to tILCs in health and disease, including transcriptomic signatures, frequency dynamics, and potential clinical value. Furthermore, we will highlight the dynamics of the NKp44+ ILC3 subset, which emerges as prognostic marker in peripheral blood for inflammatory bowel disease and leukemia. [ABSTRACT FROM AUTHOR]

• Effects of transcranial static magnetic stimulation (tSMS) are state-dependent. • tSMS increases cortical excitability if applied on an inhibited M1. • Aftereffects of tSMS originate at synaptic level. Aftereffects of non-invasive brain stimulation techniques may be brain state-dependent. Either continuous theta-burst stimulation (cTBS) as transcranial static magnetic field stimulation (tSMS) reduce cortical excitability. Our objective was to explore the aftereffects of tSMS on a M1 previously stimulated with cTBS. The interaction effect of two inhibitory protocols on cortical excitability was tested on healthy volunteers (n = 20), in two different sessions. A first application cTBS was followed by real-tSMS in one session, or sham-tSMS in the other session. When intracortical inhibition was tested with paired-pulse transcranial magnetic stimulation, LICI (ie., long intracortical inhibition) increased, although the unconditioned motor-evoked potential (MEP) remained stable. These effects were observed in the whole sample of participants regardless of the type of static magnetic field stimulation (real or sham) applied after cTBS. Subsequently, we defined a group of good-responders to cTBS (n = 9) on whom the unconditioned MEP amplitude reduced after cTBS and found that application of real-tSMS (subsequent to cTBS) increased the unconditioned MEP. This MEP increase was not found when sham-tSMS followed cTBS. The interaction of tSMS with cTBS seems not to take place at inhibitory cortical interneurons tested by LICI, since LICI was not differently affected after real and sham tSMS. Our results indicate the existence of a process of homeostatic plasticity when tSMS is applied after cTBS. This work suggests that tSMS aftereffects arise at the synaptic level and supports further investigation into tSMS as a useful tool to restore pathological conditions with altered cortical excitability. [ABSTRACT FROM AUTHOR]

[Display omitted] • Previous population genetic analysis of Opisthorchis viverrini eggs revealed two populations. • Cat- and human-specific populations use different intermediate fish hosts. • There are distinct morphologies in egg, metacercaria, and adult worm stages of O. viverrini. • Our study confirms distinct human- and cat-specific O. viverrini populations. Infection by the zoonotic fish-borne trematode, Opisthorchis viverrini, remains a crucial health issue in Thailand and neighboring countries. Recently, molecular analysis revealed two populations of putative O. viverrini : one found primarily in human hosts ("human-specific" population) and the other primarily in cats ("cat-specific" population). It is unclear how the infective stages (metacercariae) of these different populations circulate among definitive and reservoir hosts in nature. To gain an insight into this, mitochondrial cox 1 and nad 1 gene sequences of metacercariae from fish intermediate hosts were examined. None of 192 metacercariae from cyprinid fish in Lao PDR and Thailand had sequences typical of "cat-specific" O. viverrini , suggesting that cyprinid fish are not the main second intermediate hosts of this population. Interestingly, all 20 O. viverrini -like metacercariae from snakehead fish (Channa striata) shared 99.51–100% sequence identity with eggs from cats naturally infected in a previous study. Hence, we propose a modification of the known transmission dynamics of O. viverrini : consumption of metacercariae within snakehead fish provides another pathway for cats and (occasionally) humans to acquire infection. We also performed morphological comparisons of eggs, metacercariae, and adult flukes (raised in hamsters) of both Opisthorchis populations. The "cat-specific" population has eggs that are narrower and adults that are shorter and wider than in the human-specific population. The metacercaria of the "cat-specific" population is elliptical, while that of the "human-specific" population is oval, occasionally rounded. Our results confirmed that O. viverrini -like metacercariae from snakehead fish are the infective stages of the "cat-specific" fluke. This provides a new insight into the dissemination and transmission of each population in the second intermediate host. The identity of the cat-specific population is discussed. [ABSTRACT FROM AUTHOR]

• Theme 1) participants reported a lack of awareness of pregnancy-related warning signs. • Theme 2) presenting symptoms participants experienced were not typical of expected warning signs. • Theme 3) participant concerns were met with dismissal leading to delays in diagnosis. • Theme 4) participants experienced long-term sequelae from maternal sepsis. Prior studies have shown that maternal deaths due to sepsis occur due to delays in recognition, treatment, and escalation of care through medical chart reviews. This study was conducted to obtain the patient perspective for near-miss and maternal mortality cases due to sepsis. To identify quality improvement opportunities for improving maternal sepsis through patient and support person experiences. Twenty semi-structured interviews and three follow-up focus groups with patients who experienced critical illness from maternal sepsis in the United States and their support persons (when available) were conducted from May 23, 2022, through October 14, 2022. In this qualitative study, data were analyzed using inductive thematic analysis. In this qualitative study of patients with maternal sepsis and their support persons, four main quality improvement themes were identified. The themes were the following: (1) participants reported a lack of awareness of pregnancy-related warning signs and symptoms of when to seek care, (2) many of the presenting symptoms participants experienced were not typical of expected warning signs of maternal sepsis, such as severe pain, overwhelming tiredness, and lack of fever (3) participant concerns were met with dismissal leading to delays in diagnosis, (4) participants experienced long-term sequelae but had difficulty receiving screening and referrals for treatment. The findings of this study suggest that standardized patient education about the warning signs of maternal sepsis and provider education about the presentation of maternal sepsis, improved listening to patients, and follow-up for sequalae of sepsis are needed. [ABSTRACT FROM AUTHOR]

The insula and dorsal anterior cingulate cortex (dACC) are core brain regions involved in pain processing and central sensitization, a shared mechanism across various chronic pain conditions. Methods to modulate these regions may serve to reduce central sensitization, though it is unclear which target may be most efficacious for different measures of central sensitization. Objective/Hypothesis: Investigate the effect of low-intensity focused ultrasound (LIFU) to the anterior insula (AI), posterior insula (PI), or dACC on conditioned pain modulation (CPM) and temporal summation of pain (TSP). N = 16 volunteers underwent TSP and CPM pain tasks pre/post a 10 min LIFU intervention to either the AI, PI, dACC or Sham stimulation. Pain ratings were collected pre/post LIFU. Only LIFU to the PI significantly attenuated pain ratings during the TSP protocol. No effects were found for the CPM task for any of the LIFU targets. LIFU pressure modulated group means but did not affect overall group differences. LIFU to the PI reduced temporal summation of pain. This may, in part, be due to dosing (pressure) of LIFU. Inhibition of the PI with LIFU may be a future potential therapy in chronic pain populations demonstrating central sensitization. The minimal effective dose of LIFU for efficacious neuromodulation will help to translate LIFU for therapeutic options. • Low-intensity focused ultrasound (LIFU) can non-invasively target subregions of the insula and anterior cingulate. • LIFU to the posterior insula reduced pain ratings during temporal summation of pain but not conditioned pain modulation. • Estimated in vivo pressure varies across participants and brain targets and may help explain LIFU's effects. [ABSTRACT FROM AUTHOR]

Safety and efficacy are the most critical factors for the development of modern medications. For oral drugs, evaluating drug exposure under various conditions is one of the most important outcomes for clinical trials. These data will help to better understand the safety and efficacy of new drugs. Studies involving potential drug-drug interactions, proton pump inhibitors, and intake of food are often conducted to assess the above. Among the above, the influence of food on exposure to the drug is one of the key data sets for regulatory submission. Since food may have either a positive or negative effect on drug exposure, it is important to obtain an early assessment of the food effect. To better forecast and plan for clinical studies, substantial efforts have been made in the industry to develop modeling and in-vitro and in–vivo assays. Despite the efforts, predicting the effect of food on exposure without integrating the dynamic of the gastrointestinal tract in the assessment remains challenging. In this study, we evaluated the utilization of the dynamic Gastro-Intestinal Model (Tiny-TIM) for the food effect of over 20 drugs/formulations in development or on the market that covers all BCS classes. In general, the Tiny-TIM predicted food effects were in good agreement with the reported data in humans. This suggests that Tiny-TIM can successfully capture the impact of physicochemical properties on absorption under the influence of food. [ABSTRACT FROM AUTHOR]

There is conflicting evidence regarding the impact of the Coronavirus 2019 (COVID-19) pandemic on antimicrobial resistance, with few studies conducted in low- and middle-income countries. We investigated the impact of the COVID-19 pandemic on multidrug resistant organisms (MDROs) among critically ill patients and their clinical outcomes. This was a retrospective observational study of patients admitted to the medical Intensive Care Unit at Jordan University Hospital and had blood, urine, or bronchoalveolar bacterial cultures taken during 2 time periods: prepandemic (ie, 1/2016 to 1/2019) and pandemic (ie, 1/2020 to 1/2023). We compared the clinical outcomes (ie, in-hospital deaths, the need for O 2 devices, intubation, the length of hospital stay, and the occurrence of complications) and prevalence of MDROs between the 2 periods and conducted multivariate analyses. There were 1,254 patients (479 prepandemic and 775 postpandemic. The percentage of patients who had MDROs was significantly higher among patients with a culture taken during the pandemic (82.4%) compared to before it (17.6%) (P -value=.000). Multivariate analysis demonstrated that patients cultured during the pandemic were more than 3 times as likely to have an MDRO (odds ratio = 3.210; 95% confidence interval: 2.236-4.609). The increase in MDROs during the COVID-19 pandemic is an alarming threat to public health; thus, investigating the antibiotic resistance situation as the pandemic subsides is crucial. • The impact of the COVID-19 pandemic on antibiotic resistance is controversial. • Patients cultured during the pandemic were 3 times more likely to have an MDRO. • Patients with cultures during the pandemic had worse clinical outcomes. [ABSTRACT FROM AUTHOR]

Virtually everyone is exposed to power-frequency MF (50/60 Hz), inducing in our body electric fields and currents, potentially modulating brain function. MF-induced electric fields within the central nervous system can generate flickering visual perceptions (magnetophosphenes), which form the basis of international MF exposure guidelines and recommendations protecting workers and the general public. However, magnetophosphene perception thresholds were estimated 40 years ago in a small, unreplicated study with significant uncertainties and leaving open the question of the involved interaction site. We used a stimulation modality termed transcranial alternating magnetic stimulation (tAMS), delivering in situ sinusoidal electric fields comparable to transcranial alternating current stimulation (tACS). Magnetophosphene perception was quantified in 81 volunteers exposed to MF (eye or occipital exposure) between 0 and 50 mT at frequencies of 20, 50, 60 and 100 Hz. Reliable magnetophosphene perception was induced with tAMS without any scalp sensation, a major advantage as compared to tACS. Frequency-dependent thresholds were quantified using binary logistic regressions hence allowing to establish condition dependent probabilities of perception. Results support an interaction between induced current density and retinal rod cells. Beyond fundamental and immediate implications for international safety guidelines, and for identifying the interaction site underlying phosphene perception (ubiquitous in tACS experiments), our results support exploring the potential of tAMS for the differential diagnosis of retinal disorders and neuromodulation therapy. • Experimental magnetophosphene perception thresholds useful to the next international exposure limits. • Unique innovative transcranial alternating magnetic stimulation device (tAMS). The dosimerty is detailed in the manuscript. • tAMS induces in-situ electric fields equivalent to transcranial electric stimulation (tDCS and tACS), without skin sensation. [ABSTRACT FROM AUTHOR]

Early utero-placental vascular development impacts placental development and function throughout pregnancy. We investigated whether impaired first-trimester utero-placental vascular development is associated with pathologic features of the postpartum placenta. In this prospective observational study of 65 ongoing pregnancies, we obtained three-dimensional power Doppler ultrasounds of the placenta at 7, 9 and 11 weeks of gestation. We applied VOCAL software to measure placental volume (PV), virtual reality based segmentation to measure utero-placental vascular volume (uPVV) and applied a skeletonization algorithm to generate the utero-placental vascular skeleton (uPVS). Vascular morphology was quantified by assigning a morphologic characteristic to each voxel in the uPVS (i.e. end-, bifurcation-, crossing- or vessel point). Following delivery, placentas were measured and histologically examined according to the Amsterdam criteria to assess maternal vascular malperfusion (MVM). We used linear mixed models to estimate trajectories of PV, uPVV and uPVS development. Multivariable linear regression analysis with adjustments for confounders was used to evaluate associations between PV, uPVV and uPVS development and features of the postpartum placenta. We observed no associations between first-trimester PV development and measurements of the postpartum placenta. Increased first-trimester utero-placental vascular development, reflected by uPVV (β = 0.25 [0.01; 0.48]), uPVS end points (β = 0.25 [0.01; 0.48]), bifurcation points (β = 0.22 [0.05; 0.37]), crossing points (β = 0.29 [0.07; 0.52]) and vessel points (β = 0.09 [0.02; 0.17]) was positively associated with the postpartum placental diameter. uPVV was positively associated with postpartum placental weight. No associations were found with MVM. Development of the first-trimester utero-placental vasculature is associated with postpartum placental size, whereas placental tissue development contributes to a lesser extent. [Display omitted] • New imaging markers of early placental vascular development using 3D power Doppler. • Placental vascular imaging markers and histologic evaluation of the human placenta. • First-trimester vascular development is associated with postpartum placental size. [ABSTRACT FROM AUTHOR]

Noninvasive spinal stimulation has been increasingly used in research on motor control and neurorehabilitation. Despite advances in percutaneous electrical stimulation techniques, magnetic stimulation is not as commonly used as electrical stimulation. Therefore, it is still under discussion what neuronal elements are activated by magnetic stimulation of the human spinal cord. In this study, we demonstrated that transvertebral magnetic stimulation (TVMS) induced transsynaptic activation of spinal motoneuron pools in the lumbar cord. In healthy humans, paired-pulse TVMS was given over an intervertebral space between the L1-L2 vertebrae with an interpulse interval of 100 ms, and the stimulus-evoked electromyographic (EMG) responses were recorded in the lower limb muscles. The results show that the evoked EMG responses after the 2nd pulse were clearly suppressed compared with the widespread responses evoked after the 1st pulse in the muscles of the lower extremity, indicating that the transsynaptic activation of spinal motoneurons by the 2nd pulse was suppressed by the effects produced by the 1st pulse. The inconsistent modulation of response suppression to stimulus intensity across individuals suggests that the TVMS-evoked EMG responses are composed of the compound potentials mediated by the direct activation of motor axons and the transsynaptic activation of motoneuron pools through sensory afferents and that the recruitment order of those fibers by TVMS may be nonhomogeneous across individuals. • This study aimed to determine which neuronal elements are activated by transvertebral magnetic stimulation (TVMS) in humans. • Paired-pulse TVMS induced suppression of the muscle response after the 2nd stimulation. • TVMS induced transsynaptic activation of spinal motoneuron pools. [ABSTRACT FROM AUTHOR]

Computed tomography (CT) imaging has become indispensable in the management of medical oncology patients. Risks associated with high cumulative effective dose (CED) are relevant in testicular cancer patients. Split-bolus protocols, whereby the contrast medium injection is divided into two, followed by combining the required phase images in a single scan acquisition has been shown to provide images of comparable image quality and less radiation dose compared to single-bolus split-phase CT for various indications. We retrospectively evaluated the performance of split-bolus and single-bolus protocols in patients having follow-up CT imaging for testicular cancer surveillance. 45 patients with testicular cancer undergoing surveillance CT imaging of the thorax, abdomen, and pelvis who underwent split-bolus and single-bolus protocols were included. Quantitative image quality analysis was conducted by placing region of interests in pre-defined anatomical sub-structures within the abdominal cavity. The signal-to-noise ratio (SNR) and radiation dose in the form of dose length product (DLP) and effective dose (ED) were recorded. The DLP and ED for the single-bolus, split-phase acquisition was 506 ± 89 mGy cm and 7.59 ± 1.3 mSv, respectively. For the split-bolus, single-phase acquisition, 397 ± 94 mGy∗cm and 5.95 ± 1.4 mSv, respectively (p < 0.000). This represented a 21.5 % reduction in radiation dose exposure. The SNR for liver, muscle and fat for the single-bolus were 7.4, 4.7 and 8, respectively, compared to 5.5, 3.8 and 7.4 in the split-bolus protocol (p < 0.001). In a testicular cancer patient cohort undergoing surveillance CT imaging, utilization of a split-bolus single-phase acquisition CT protocol enabled a significant reduction in radiation dose whilst maintaining subjective diagnostic acceptability. Implications for Practice: Use of split-bolus, single-phase acquisition has the potential to reduce CED in surveillance of testicular cancer patients. [ABSTRACT FROM AUTHOR]

Our group previously reported a burn biopsy algorithm (BBA-V1) for categorizing burn wound depth. Here, we sought to promulgate a newer, simpler version of the BBA (BBA-V2). Burn wounds undergoing excision underwent 4 mm biopsies procured every 25 cm2. Serial still photos were obtained at enrollment and at excision intraoperatively. Burn wounds assessed as likely to heal by 21 days were imaged within 72 h of injury and at 21 days. A sample of 798 burn wound biopsies were classified by both BBAV1 and BBAV2 algorithms. For nonoperative burn wounds, the proportion of healing versus nonhealing pixels at 21 days after injury were compared. The 798 biopsies were classified by BBAV1 as 24% SPT, 47% DPT, 28% FT and by BBAV2 as 3% SPT, 67% DPT, and 30% FT (p < 0.0001). Overall, the proportion of biopsies whose wound reclassification changed from a nonoperative to operative pathway was 21% (95% CI: 18–24%). Nonoperative wounds judged at injury as being SPT contained 12.8 million pixels. Repeat 21-day imaging revealed 11.3 million healed pixels (accuracy = 89.6% (95% CI: 89.59–89.62)). BBA-V2 was associated with a significantly higher concordance with visual assessment for burn wounds clinically judged as deep partial and full thickness. • Burn surgeons clinically diagnosed burn wounds as DPT or FT with 97% accuracy. • Burn surgeons predicted spontaneous healing with 89% accuracy. • Our collection of 1192 burn wound biopsies is the largest in the literature. [ABSTRACT FROM AUTHOR]

Type 3 innate lymphoid cells (ILC3) are important in tissue homeostasis. In the gut, ILC3 repair damaged epithelium and suppress inflammation. In allogeneic hematopoietic cell transplantation (HCT), ILC3 protect against graft-versus-host disease (GvHD), most likely by restoring tissue damage and preventing inflammation. We hypothesize that supplementing HCT grafts with interleukin-22 (IL-22)-producing ILC3 may prevent acute GvHD. We therefore explored ex vivo generation of human IL-22-producing ILC3 from hematopoietic stem and progenitor cells (HSPC) obtained from adult, neonatal and fetal sources. We established a stroma-free system culturing human cord blood-derived CD34+ HSPC with successive cytokine mixes for 5 weeks. We analyzed the presence of phenotypically defined ILC, their viability, proliferation and IL-22 production (after stimulation) by flow cytometry and enzyme-linked immunosorbent assay (ELISA). We found that the addition of recombinant human IL-15 and the enhancer of zeste homolog 1/2 inhibitor UNC1999 promoted ILC3 generation. Similar results were demonstrated when UNC1999 was added to CD34+ HSPC derived from healthy adult granulocyte colony-stimulating factor mobilized peripheral blood and bone marrow, but not fetal liver. UNC1999 did not negatively impact IL-22 production in any of the HSPC sources. Finally, we observed that autologous HSPC mobilized from the blood of adults with hematological malignancies also developed into ILC3, albeit with a significantly lower capacity. Together, we developed a stroma-free protocol to generate large quantities of IL-22-producing ILC3 from healthy adult human HSPC that can be applied for adoptive transfer to prevent GvHD after allogeneic HCT. [Display omitted] [ABSTRACT FROM AUTHOR]

2D and 3D in vitro models can be used to assess human immune responses to vaccination. Human whole-blood assays can be used as 2D in vitro tools to assess human innate and adaptive immune responses. 3D tissue models can recreate the complexity of lymph nodes. Tonsils can be used to model peripheral lymphoid organs and organoids. Skin tissue and capillary vein models can be used to investigate human immunity in vitro. High-density cell in vitro models can facilitate lymph node follicle formation. Human in vitro modeling is an emerging approach for preclinical evaluation of candidate vaccines with certain key advantages over animal models in accounting for species specificity and enabling direct benchmarking in the same study participant, while accounting for demographic variables such as age, sex, and comorbidity. 2D models offering simplicity and practicality and microphysiological 3D models that consider tissue architecture can model innate and adaptive immune responses and can accelerate and de-risk vaccine discovery and development. The human immune system is a complex network of coordinated components that are crucial for health and disease. Animal models, commonly used to study immunomodulatory agents, are limited by species-specific differences, low throughput, and ethical concerns. In contrast, in vitro modeling of human immune responses can enable species- and population-specific mechanistic studies and translational development within the same study participant. Translational accuracy of in vitro models is enhanced by accounting for genetic, epigenetic, and demographic features such as age, sex, and comorbidity. This review explores various human in vitro immune models, considers evidence that they may resemble human in vivo responses, and assesses their potential to accelerate and de-risk vaccine discovery and development. The human immune system is a complex network of coordinated components that are crucial for health and disease. Animal models, commonly used to study immunomodulatory agents, are limited by species-specific differences, low throughput, and ethical concerns. By contrast, in vitro modeling of human immune responses can enable species- and population-specific mechanistic studies and translational development within the same study participant. Translational accuracy of in vitro models is enhanced by accounting for genetic, epigenetic, and demographic features such as age, sex, and comorbidity. This review explores various human in vitro immune models, considers evidence that they may resemble human in vivo responses, and assesses their potential to accelerate and de-risk vaccine discovery and development. [ABSTRACT FROM AUTHOR]

• A Markov model provides lifetime estimates for HRQoL and health care costs. • Early surgery is cost effective from a health care payer perspective. • Early surgery for mild DCM provides improved lifetime HRQoL. Degenerative cervical myelopathy (DCM) is a form of acquired spinal cord compression and contributes to reduced quality of life secondary to neurological dysfunction and pain. There remains uncertainty regarding optimal management for individuals with mild myelopathy. Specifically, owing to lacking long-term natural history studies in this population, we do not know whether these individuals should be treated with initial surgery or observation. We sought to perform a cost-utility analysis to examine early surgery for mild degenerative cervical myelopathy from the healthcare payer perspective. We utilized data from the prospective observational cohorts included in the Cervical Spondylotic Myelopathy AO Spine International and North America studies to determine health related quality of life estimates and clinical myelopathy outcomes. We recruited all patients that underwent surgery for DCM enrolled in the Cervical Spondylotic Myelopathy AO Spine International and North America studies between December 2005 and January 2011. Clinical assessment measures were obtained using the Modified Japanese Orthopedic Association scale and health-related quality of life measures were obtained using the Short Form-6D utility score at baseline (preoperative), 6 months, 12 months and 24 months postsurgery. Cost measures inflated to January 2015 values were obtained using pooled estimates from the hospital payer perspective for surgical patients. We employed a Markov state transition model with Monte Carlo microsimulation using a lifetime horizon to obtain an incremental cost utility ratio associated with early surgery for mild myelopathy. Parameter uncertainty was assessed through deterministic means using one-way and two-way sensitivity analyses and probabilistically using parameter estimate distributions with microsimulation (10,000 trials). Costs and utilities were discounted at 3% per annum. Initial surgery for mild degenerative cervical myelopathy was associated with an incremental lifetime increase of 1.26 quality-adjusted life years (QALY) compared to observation. The associated cost incurred to the healthcare payer over a lifetime horizon was $12,894.56, resulting in a lifetime incremental cost-utility ratio of $10,250.71/QALY. Utilizing a willingness to pay threshold in keeping with the World Health Organization definition of "very cost-effective" ($54,000 CDN), the probabilistic sensitivity analysis demonstrated that 100% of cases were cost-effective. Surgery compared to initial observation for mild degenerative cervical myelopathy was cost-effective from the Canadian healthcare payer perspective and was associated with lifetime gains in health-related quality of life. [ABSTRACT FROM AUTHOR]

The subcutaneous implantable cardioverter-defibrillator (S-ICD) delivers 80 J shocks from an 8 cm left-parasternal coil to a 59 cm3 left lateral pulse generator (PG). A system that defibrillates with lower energy could significantly reduce PG size. Computer modeling and animal studies suggested that a second shock coil either parallel to the left-parasternal coil or transverse from the xiphoid to the PG pocket would significantly reduce the defibrillation threshold. The purpose of this study was to acutely assess the defibrillation efficacy of parallel and transverse configurations in patients receiving an S-ICD. Testing was performed in patients receiving a conventional S-ICD system. Success at 65 J was required before investigational testing. A second electrode was temporarily inserted from the xiphoid incision connected to the PG with an investigational Y-adapter. Phase 1 (n = 11) tested the parallel configuration. Phase 2 (n = 21) tested both parallel and transverse configurations in random order. This study enrolled 35 patients (28 males (80%); mean age 51 ± 17 years; left ventricular ejection fraction 40% ± 15%; body mass index 26 ± 4 kg/m2; prior myocardial infarction 46%; congestive heart failure 49%; cardiomyopathy 63%). Compared to the conventional S-ICD system, mean shock impedance decreased for both parallel (69 ± 15 Ω vs 86 ± 20 Ω; n = 33; P <.001) and transverse (56 ± 14 Ω vs 81 ± 21 Ω; n = 20; P <.001) configurations. Shock success rates at 20, 30, and 40 J were 55%, 79%, 97%, and 25%, 70%, 90% for parallel and transverse configurations, respectively. Defibrillation threshold testing was well tolerated with no serious adverse events. Adding a second shock coil, particularly in the parallel configuration, significantly reduced the impedance and had a high likelihood of defibrillation success at energies ≤40 J. This may enable the development of a smaller S-ICD. [ABSTRACT FROM AUTHOR]

• iTEPs reflect both direct (electromagnetic) and indirect (acoustic) effects on iEEG. • TMS clicks induce auditory evoked potentials that can obscure signals of interest. • TAAC sound masking software effectively masks the TMS click. • TAAC prevents TMS auditory evoked potentials in iEEG auditory cortex electrodes. • Auditory masking changes the morphology of iTEPs outside of auditory regions. [ABSTRACT FROM AUTHOR]

We set up this study to understand the underlying mechanisms of reduced ceramides on immune cells in acute rejection (AR). The concentrations of ceramides and sphingomyelins were measured in the sera from hepatic transplant patients, skin graft mice and hepatocyte transplant mice by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Serum concentrations of C24 ceramide, C24:1 ceramide, C16:0 sphingomyelin, and C18:1 sphingomyelin were lower in liver transplantation (LT) recipients with than without AR. Comparisons with the results of LT patients with infection and cardiac transplant patients with cardiac allograft vasculopathy in humans and in mouse skin graft and hepatocyte transplant models suggested that the reduced C24 and C24:1 ceramides were specifically involved in AR. A ceramide synthase inhibitor, fumonisin B1 exacerbated allogeneic immune responses in vitro and in vivo, and reduced tolerogenic dendritic cells (tDCs), while increased P3-like plasmacytoid DCs (pDCs) in the draining lymph nodes from allogeneic skin graft mice. The results of mixed lymphocyte reactions with ceranib-2, an inhibitor of ceramidase, and C24 ceramide also support that increasing ceramide concentrations could benefit transplant recipients with AR. The results suggest increasing ceramides as novel therapeutic target for AR, where reduced ceramides were associated with the changes in DC subsets, in particular tDCs. [ABSTRACT FROM AUTHOR]

Acinar-to-ductal metaplasia (ADM) has been shown to contribute to the development of pancreatic ductal adenocarcinoma (PDAC) in genetically engineered mouse models, but little is known about whether acinar cell plasticity contributes to carcinogenesis in human PDAC. We aimed to assess whether cancer cells that stain positive for amylase and CK19 (ADM-like cancer cells) are present in human resected PDAC and to investigate their role in tumor progression. We immunohistochemically investigated the presence of ADM-like cancer cells, and compared the clinical and histological parameters of PDAC patients with and without ADM-like cancer cells. ADM-like cancer cells were detected in 16 of 60 (26.7%) PDAC specimens. Positive staining for anterior gradient protein 2 (AGR2) was observed in 14 of 16 (87.5%) PDAC specimens with ADM-like cancer cells. On the other hand, the intensity of AGR2 expression (negative, low/moderate or high) was lower in PDAC with ADM-like cancer cells (9/7) than in PDAC without these cells (11/33) (P = 0.032). The presence of ADM-like cancer cells was significantly correlated with increased cell proliferation (P = 0.012) and tended to be associated with MUC1 expression (P = 0.067). These results indicated that acinar cells may act as the origin of human PDAC, and that their presence may be useful for the stratification of human PDAC to predict prognosis. [ABSTRACT FROM AUTHOR]

Intervertebral disc degeneration (IVDD) is an incurable, specific treatment-orphan disease with an increasing burden worldwide. Although great efforts have been made to develop new regenerative therapies, their clinical success is limited. Characterize the metabolomic and gene expression changes underpinning human disc degeneration. This study also aimed to disclose new molecular targets for developing and optimizing novel biological approaches for IVDD. Intervertebral disc cells were obtained from IVDD patients undergoing circumferential arthrodesis surgery or from healthy subjects. Mimicking the harmful microenvironment of degenerated discs, cells isolated from the nucleus pulposus (NP) and annulus fibrosus (AF) were exposed to the proinflammatory cytokine IL-1β and the adipokine leptin. The metabolomic signature and molecular profile of human disc cells were unraveled for the first time. The metabolomic and lipidomic profiles of IVDD and healthy disc cells were analyzed by high-performance liquid chromatography-mass spectrometry (UHPLC-MS). Gene expression was investigated by SYBR green-based quantitative real-time RT-PCR. Altered metabolites and gene expression were documented. Lipidomic analysis revealed decreased levels of triacylglycerols (TG), diacylglycerol (DG), fatty acids (FA), phosphatidylcholine (PC), lysophosphatidylinositols (LPI) and sphingomyelin (SM), and increased levels of bile acids (BA) and ceramides, likely promoting disc cell metabolism changing from glycolysis to fatty acid oxidation and following cell death. The gene expression profile of disc cells suggests LCN2 and LEAP2/GHRL as promising molecular therapeutic targets for disc degeneration and demonstrates the expression of genes related to inflammation (NOS2, COX2, IL-6, IL-8, IL-1β , and TNF-α) or encoding adipokines (PGRN, NAMPT, NUCB2, SERPINE2 , and RARRES2), matrix metalloproteinases (MMP9 and MMP13), and vascular adhesion molecules (VCAM1). Altogether, the presented results disclose the NP and AF cell biology changes from healthy to degenerated discs, allowing the identification of promising molecular therapeutic targets for intervertebral disc degeneration. Our results are relevant to improving current biological-based strategies aiming to repair IVD by restoring cellular lipid metabolites as well as adipokines homeostasis. Ultimately, our results will be valuable for successful, long-lasting relief of painful IVDD. [ABSTRACT FROM AUTHOR]

Testing of dental materials when in contact with innate immune cells has been so far hindered by the lack of proper in vitro models. Human primary monocyte-derived macrophages (MDMs) would be an excellent option to this aim. However, the inability to detach them from the tissue culture plates contrast the possibility to culture them on biomaterials. The goal of the present work is to present and validate an innovative protocol to obtain MDMs from peripheral blood monocytes, and to reseed them in contact with biomaterials without altering their viability and phenotype. We differentiated MDMs on ultra-low attachment tissue culture plastics and recovered them with specific detachment solution in order to be reseeded on a secondary substrate. Therefore, using biological assays (RT-PCR, Western blot, and immunofluorescence) we compared their phenotype to MDMs differentiated on standard culture plates. Transferred MDMs keep their differentiated M0 resting state, as well as the ability to be polarized into M1 (pro-inflammatory) or M2 (anti-inflammatory) macrophages. These data provide the dental material research community the unprecedented possibility to investigate the immunomodulatory properties of biomaterials for dental application. [ABSTRACT FROM AUTHOR]

The future health of the offspring can be influenced by longstanding maternal anxiety and depression disorders during pregnancy. The present study aimed to explore the effect of psychiatric disorders during pregnancy on placental epigenetics. We measured DNA methylation patterns in term-placentas of women either suffering longstanding anxiety and depression symptoms (Index group, with overt symptoms), or a healthy population (Control, none/only mild symptoms). Whole genome DNA methylation profiling was performed using the TruSeq® Methyl Capture EPIC Library Prep Kit (Illumina, San Diego, CA, USA) for library preparation and NGS technology for genomic DNA sequencing. The results of high-throughput DNA methylation analysis revealed that the Index group had differential DNA methylation at epigenome-wide significance (p < 0.05) in 226 genes in the placenta. Targeted enrichment analyses identified hypermethylation of genes associated with psychiatric disorders (BRINP1 , PUM1), and ion homeostasis (COMMD1), among others. The ECM (extracellular matrix)-receptor interaction pathway was significantly dysregulated in the Index group compared to the Control. In addition, DNA methylation/mRNA integration analyses revealed that four genes with key roles in neurodevelopment and other important processes (EPB41L4B, BMPR2, KLHL18, and UBAP2) were dysregulated at both, DNA methylation and transcriptome levels in the Index group compared to Control. The presented results increase our understanding of how maternal psychiatric disorders may affect the newborn through placental differential epigenome, suggesting DNA methylation status as a biomarker when aiming to design new preventive techniques and interventions. • Pregnancy anxiety and depression may compromise the health of the newborn. • Maternal psychiatric disorders affect placental DNA methylation status. • Dysregulated placental DNA methylation may alter gene function. [ABSTRACT FROM AUTHOR]

Several single cell profiling studies of human hematopoietic stem and progenitor cells indicate the existence of circulating CD123+CD127+ lymphoid progenitors that express IRF8 and which are dendritic cell (DC)- and lymphoid [B and T cell/innate lymphoid cell (ILC)]-transcriptionally primed. IRF8 -expressing CD123+CD127+ lymphoid progenitors are also found in tissues that support ILC, DC, and T cell development, suggesting that downstream lineage specification and commitment are modulated by the specific tissue microenvironment. Expression of IRF8 unlocks DC lineage potential in these human lymphoid progenitors but is differentially regulated in downstream steps of DC and B/T/ILC differentiation. CD123 expression (a surrogate cell surface marker for IRF8) does not preclude hematopoietic progenitors from harboring lymphoid potential. The abundance of CD123+CD127+ lymphoid progenitors in peripheral blood is affected by stem cell mobilizers. This might impact lymphoid reconstitution following transplantation with hematopoietic stem cell-mobilized peripheral blood. The hematopoietic model proposed here highlights an underappreciated role for transcription factor interferon regulatory factor 8 in unlocking human dendritic cell, B and T cell, and innate lymphoid cell potential in CD123+CD127+ lymphoid progenitors. Understanding the development and differentiation of CD123+CD127+ lymphoid progenitors across tissues has important basic and translational implications. Lymphopoiesis is the process in which B and T cells, and innate lymphoid cells (ILCs) develop from hematopoietic progenitors that exhibit early lymphoid priming. The branching points where lymphoid-primed human progenitors are further specified to B/T/ILC differentiation trajectories remain unclear. Here, we discuss the emerging role of interferon regulatory factor (IRF)8 as a key factor to bridge human lymphoid and dendritic cell (DC) differentiation, and the current evidence for the existence of circulating and tissue-resident CD123+CD127+ lymphoid progenitors. We propose a model whereby DC/B/T/ILC lineage programs in circulating CD123+CD127+ lymphoid progenitors are expressed in balance. Upon tissue seeding, the tissue microenvironment tilts this molecular balance towards a specific lineage, thereby determining in vivo lineage fates. Finally, we discuss the translational implication of these lymphoid precursors. [ABSTRACT FROM AUTHOR]

• Growing numbers of monogenetic diseases reveal distinct requirement in human and mouse B lymphopoiesis. • Corresponding stages of B cell development are defined by different markers in mouse and humans. • Limited transferability of mouse models to human point mutations in defined domains of transcription factors driving B cell fate. • Cytokine signals instruct B cell fate with distinct requirements in mouse and humans. Early B cell development in the bone marrow ensures the replenishment of the peripheral B cell pool. Immature B cells continuously develop from hematopoietic stem cells, in a process guided by an intricate network of transcription factors as well as chemokine and cytokine signals. Humans and mice possess somewhat similar regulatory mechanisms of B lymphopoiesis. The continuous discovery of monogenetic defects that impact early B cell development in humans substantiates the similarities and differences with B cell development in mice. These differences become relevant when targeted therapeutic approaches are used in patients; therefore, predicting potential immunological adverse events is crucial. In this review, we have provided a phenotypical classification of human and murine early progenitors and B cell stages, based on surface and intracellular protein expression. Further, we have critically compared the role of key transcription factors (Ikaros, E2A, EBF1, PAX5, and Aiolos) and chemo- or cytokine signals (FLT3, c-kit, IL-7R, and CXCR4) during homeostatic and aberrant B lymphopoiesis in both humans and mice. [ABSTRACT FROM AUTHOR]

In aged animals from worms to humans, transcriptional elongation rates are faster, leading to changes in transcript quality and alternative splicing. Recent work by Debès et al. shows how interventions that slow elongation rates, such as mutating RNA polymerase II (Pol II) or increasing nucleosome density to impose transcriptional 'traffic', delay senescence and promote longevity. In aged animals from worms to humans, transcriptional elongation rates are faster, leading to changes in transcript quality and alternative splicing. Recent work by Debès et al. shows how interventions that slow elongation rates, such as mutating RNA polymerase II (Pol II)or increasing nucleosome density to impose transcriptional 'traffic', delay senescence and promote longevity. [ABSTRACT FROM AUTHOR]

• Basement membrane ageing is thought to result from damage accumulation in long-lived matrix proteins, in turn contributing to declining kidney function. • An emerging proteomic analysis technique, peptide location fingerprinting, was applied to young and aged human kidney glomerular and tubulointerstitial mass spectrometry datasets to identify basement membrane proteins with ageing-modified structures. • Alterations in peptide yields were located within functional regions of key basement membrane components, including the HS-binding domain of agrin, integrin-binding regions of laminins 521 and 511 and canstatin- and endostatin-releasing NC1 domains of collagens IV and XVIII. • Periostin and the collagen IV α2 exhibited age-dependent differences that were highly conserved between human kidney and mouse lung, suggesting that similar basement membrane ageing processes occur across species and organs. During ageing, the glomerular and tubular basement membranes (BM) of the kidney undergo a progressive decline in function that is underpinned by histological changes, including glomerulosclerosis and tubular interstitial fibrosis and atrophy. This BM-specific ageing is thought to result from damage accumulation to long-lived extracellular matrix (ECM) protein structures. Determining which BM proteins are susceptible to these structure-associated changes, and the possible mechanisms and downstream consequences, is critical to understand age-related kidney degeneration and to identify markers for therapeutic intervention. Peptide location fingerprinting (PLF) is an emerging proteomic mass spectrometry analysis technique capable of identifying ECM proteins with structure-associated differences that may occur by damage modifications in ageing. Here, we apply PLF as a bioinformatic screening tool to identify BM proteins with structure-associated differences between young and aged human glomerular and tubulointerstitial compartments. Several functional regions within key BM components displayed alterations in tryptic peptide yield, reflecting potential age-dependent shifts in molecular (e.g. laminin-binding regions in agrin) and cellular (e.g. integrin-binding regions in laminins 521 and 511) interactions, oxidation (e.g. collagen IV) and the fragmentation and release of matrikines (e.g. canstatin and endostatin from collagens IV and XVIII). Furthermore, we found that periostin and the collagen IV α2 chain exhibited structure-associated differences in ageing that were conserved between human kidney and previously analysed mouse lung, revealing BM components that harbour shared susceptibilities across species and organs. [ABSTRACT FROM AUTHOR]

Human teeth are composed mainly of dentin, formed by the odontoblasts. Dentin matrix protein 1 (DMP1) is one of odontoblast differentiation's most important growth factors. Human DMP1 has yet to be completely identified or studied. This study aimed to clone and characterize human DMP1. The DMP1 gene sequence was prepared and cloned by transfection of human 293 cells. The recombinant DMP1 was purified and characterized. The results suggested its future use in dental tissue regeneration and tissue engineering. [ABSTRACT FROM AUTHOR]

Single-element low-intensity focused ultrasound (LIFU) is an emerging form of human neuromodulation. Current coupling methods are impractical for clinical bedside use. Here, we evaluate commercially available high-viscosity gel polymer matrices as couplants for human LIFU neuromodulation applications. We first empirically tested the acoustic transmission of three densities at 500 kHz and then subjected the gel with the least acoustic attenuation to further tests of the effect of thickness, frequency, de-gassing and production variability. The highest-density gel had the lowest acoustic attenuation (3.3%) with low lateral (<0.5 mm) and axial (<2 mm) beam distortion. Different thicknesses of the gel up to 10 mm did not appreciably affect results. The gel polymers exhibited frequency-dependent attenuation at 1 and 3 MHz up to 86.6%, as well as significant beam distortion >4 mm. Poor de-gassing methods also increased pressure attenuation at 500 kHz up to 59.6%. Standardized methods of making these gels should be established to reduce variability. Commercially available de-gassed, high-density gel matrices are a low-cost, easily malleable, low-attenuation and distortion medium for the coupling of single-element LIFU transducers for human neuromodulation applications at 500 kHz. [ABSTRACT FROM AUTHOR]

• Leishmania (Mundinia) macropodum causes cutaneous leishmaniasis in marsupials. • Enzyme-linked immunosorbent assay showed that 20.57% of humans (n = 282) were immunoglobulin (Ig) G positive for L. macropodum. • Samples with L. macropodum IgG had IgG1 and IgG2 as the prevalent subclasses. • We show that humans have evidence of L. macropodum exposure. • Future studies should investigate whether L. macropodum can viably infect humans. The Australian Leishmania (Mundinia) macropodum parasite causes cutaneous leishmaniasis among marsupial species. Although cutaneous leishmaniasis is a major public health burden worldwide, it is not clear if humans are naturally exposed to the unique L. macropodum. To assess whether humans have an immunoglobulin (Ig) G response to L. macropodum , we examined anti- Leishmania antibodies among humans residing in a region of marsupial Leishmania endemicity in Australia. Using a serological enzyme-linked immunosorbent assay, we characterized Leishmania -specific IgG and IgG subclass responses to soluble Leishmania antigen from L. macropodum, and other Leishmania species (L. donovani, L. major, and L. mexicana) in 282 blood donor samples. We found that 20.57% of individuals demonstrated a positive total IgG response to L. macropodum. For individuals with antibodies to soluble Leishmania antigen from one Leishmania species, there was no increased likelihood of recognition to other Leishmania species. For samples with detectable L. macropodum IgG, IgG1 and IgG2 were the prevalent subclasses detected. It is not yet clear whether the IgG antibody detection in this study reflects exposure to Leishmania parasites or a cross-reactive immune response that was induced against an unrelated immunogen. Future studies should investigate whether L. macropodum can result in a viable infection in humans. [Display omitted] [ABSTRACT FROM AUTHOR]

Background: The fermentation of undigested material in the ileum is quantitatively important. However, the respective contributions of the microbial composition and the substrate to ileal fermentation are unclear. Objective: This aim was to investigate the contribution of microbial composition and fiber source to in vitro ileal fermentation outcomes. Methods: Thirteen ileal cannulated female pigs (Landrace/Large White; 9-wk-old; 30.5 kg body weight) were given diets containing black beans, wheat bread, chickpeas, peanuts, pigeon peas, sorghum, or wheat bran as the sole protein source for 7 d (100 g protein/kg dry matter diet). On day 7, ileal digesta were collected and stored at -80°C for microbial analysis and in vitro fermentation. For each diet, a pooled ileal inoculum was prepared to ferment different fiber sources (cellulose, pectin, arabinogalactan, inulin, fructooligosaccharides, and resistant starch) for 2 h at 37°C. Organic matter fermentability and organic acid production were determined following in vitro fermentation. Data were analyzed using a 2-way ANOVA (inoculum x fiber). Results: Forty-five percent of the identified genera in the digesta differed across diets. For instance, the number of Lactococcus was 115-fold greater (P ≤ 0.05) in the digesta of pigs fed the pigeon pea diet than for pigs fed the wheat bran diet. For both in vitro organic matter fermentability and organic acid production, there were significant (P ≤ 0.05)n interactions between the inoculum and the fiber source. For instance, pectin and resistant starch resulted in 1.6- to 31-fold more (P ≤ 0.05) lactic acid production when fermented by the pigeon pea inoculum than other inocula. For specific fiber sources, statistically significant correlations were found between the number of bacteria from certain members of the ileal microbial community and fermentation outcomes. Conclusions: Both the fiber source fermented and the ileal microbial composition of the growing pig affected in vitro fermentation; however, the effect of the fiber source was predominant. [ABSTRACT FROM AUTHOR]

• Participants' perceptual decisions were modulated by observing others' decisions. • The computational model could deduce the participants' decisions. • Model-based analysis revealed that MPFC was associated with self-other discrepancy. • Parietal region showed altered activity patterns after observing others' decisions. Our perceptions and decisions are often implicitly influenced by observing another's actions. However, it is unclear how observing other people's perceptual decisions without interacting with them can engage the processing of self-other discrepancies and change the observer's decisions. In this study, we employed functional magnetic resonance imaging and a computational model to investigate the neural basis of how unilaterally observing the other's perceptual decisions modulated one's own decisions. The experimental task was to discriminate whether the number of presented dots was higher or lower than a reference number. The participants performed the task solely while unilaterally observing the performance of another "participant," who produced overestimations and underestimations in the same task in separate sessions. Results of the behavioral analysis showed that the participants' decisions were modulated to resemble those of the other. Image analysis based on computational model revealed that the activation in the medial prefrontal cortex was associated with the discrepancy between the inferred participant's and the presented other's decisions. In addition, the number-sensitive region in the superior parietal region showed altered activation patterns after observing the other's overestimations and underestimations. The activity of the superior parietal region was not involved in assessing the observation of other's perceptual decisions, but it was engaged in plain numerosity perception. These results suggest that computational modeling can capture the neuro-behavioral processing of self-other discrepancies in perception followed by the activity modulation in the number-sensitive region in the task of dot-number estimation. [ABSTRACT FROM AUTHOR]

Single-cell transcriptomic (scRNA-seq) and proteomic [cytometry time of flight (CyTOF) mass spectrometry] approaches have recently been applied to cells isolated from Dupuytren's nodules and have enabled us to comprehensively profile the complex cellular ecosystem within Dupuytren's nodules. This has highlighted the high degree of heterogeneity of both fibroblast and myofibroblast cell states. In silico trajectory analysis has facilitated the identification of myofibroblast precursors such as pericytes and resident fibroblasts. Differential gene analysis along the differentiation trajectory identified novel markers of pathogenic myofibroblasts, including CD82, podoplanin, TNFSF12A, and the transcription factors SCX and DMRT2. Interleukin 13 (IL-13) has recently been shown to have a role in Dupuytren's disease, where it is expressed by mast cells, and enhances the expression of profibrotic gene expression, which was sensitive to inhibition by the JAK inhibitor, tofacitinib. Tumour necrosis factor (TNF) has also been shown to be produced by both M2 macrophages and mast cells in DD, and chronic expression was maintained by a positive feedback loop of IL-33. TNF signals through TNFR2 to upregulate the expression of α-smooth muscle actin (α-SMA) and collagen. These findings led to the assessment of local injection of anti-TNF (adalimumab) into the nodules in a Phase 2b clinical trial, which met the primary endpoint of reduction in nodule hardness and the secondary endpoint of nodule size, suggesting the potential to control the progression of early-stage Dupuytren's disease. We review the biology of Dupuytren's disease (DD), a common localised fibrotic disorder of the hand. The disease develops through a complex interplay of genetic and environmental factors, and epigenetic signalling. The early-stage disease nodules comprise a complex milieu of stromal and immune cells which interact to promote disease development. Recently, inhibition of tumour necrosis factor (TNF) locally resulted in softening and a decrease in nodule size, potentially controlling disease progression. Unlike fibrotic disorders of the visceral organs, the easy access to tissue in DD patients enables dissection of the cellular landscape and molecular signalling pathways. In addition, the study of DD may have wider benefits in enhancing our understanding of less-accessible fibrotic tissues. [ABSTRACT FROM AUTHOR]

Injury to the nervous system can generate chronic pain driven by persistent hyperactivity of nociceptors, sensory neurons specialized to detect damaging stimuli. Recent evidence argues against the near-universal assumption that persistent nociceptor hyperactivity and resulting chronic pain are always maladaptive. Widespread conservation of mechanisms of nociceptor function and hyperactivity are well documented, as well as distinct adaptations to divergent nociceptive needs. Persistent nociceptor hyperactivity could have evolved to drive painful hypervigilance during persistent physical impairment, enhancing survival by decreasing subsequent risk of predatory or aggressive attack. Differences between rodent and human nociceptors are consistent with the possibility that, during relatively recent evolution, nociceptors increased their susceptibility to persistent hyperactivity, increasing the propensity of humans to chronic pain. Chronic pain caused by injury or disease of the nervous system (neuropathic pain) has been linked to persistent electrical hyperactivity of the sensory neurons (nociceptors) specialized to detect damaging stimuli and/or inflammation. This pain and hyperactivity are considered maladaptive because both can persist long after injured tissues have healed and inflammation has resolved. While the assumption of maladaptiveness is appropriate in many diseases, accumulating evidence from diverse species, including humans, challenges the assumption that neuropathic pain and persistent nociceptor hyperactivity are always maladaptive. We review studies indicating that persistent nociceptor hyperactivity has undergone evolutionary selection in widespread, albeit selected, animal groups as a physiological response that can increase survival long after bodily injury, using both highly conserved and divergent underlying mechanisms. [ABSTRACT FROM AUTHOR]

Infantile hypertrophic pyloric stenosis (IHPS) is one of the most common diseases that require surgical intervention amongst the paediatric population. Although the treatment and the diagnosis of pyloric stenosis are well established, the perinatal risk factors associated with it still need further investigation. We searched the following databases: Cochrane, Google Scholar, PubMed, and Scopus. Studies were included if they were case-control or cohort in design and studied the perinatal risk factors associated with IHPS. The quality of the included studies was assessed using the Newcastle-Ottawa scale (NOS). Twenty-one articles were included in this meta-analysis, including 18,104,753 participants. Our analysis showed a significant association between IHPS and male sex (RR=2.71, 95% CI:1.93–3.78), maternal smoking (RR=1.75, 95% CI: 1.54 - 2.00), bottle-feeding (RR=1.68, 95% CI: 1.42 - 1.98), being first born (RR=1.23, 95% CI:1.07–1.40), African ethnicity (RR=0.51, 95% CI: 0.35–0.75), and cesarean section (RR=1.57, 95% CI: 1.49–1.66). On the contrary, there was no significant association between IHPS and multiple gestations, preterm labour, being born in summer, and small for gestational age (SGA). In conclusion, our analysis showed that male sex, bottle feeding, maternal smoking and African ethnicity were significantly associated with the risk of IHPS. However, most of the included articles were retrospective in design which necessitates conducting future prospective well-designed studies to further investigate the risk factors of IHPS. [ABSTRACT FROM AUTHOR]

Bone, platelet concentrate, and tooth-derived dentin/cementum have been used as autologous materials in regenerative medicine Dentin materials were first recycled in 2002 for bone regeneration in humans, although bone autografts were noted in the 19th century, and auto-platelet concentrates were developed in 1998. Dentin/cementum-based material therapy has been applied as an innovative technique for minimally invasive bone surgery, while bone autografts are associated with donor site morbidity. In October 2021, PubMed, Google Scholar, Scopus, and the Cochrane Library databases from 1980 to 2020 were screened. The demineralized dentin/cementum matrix (DDM) had better performance in bone induction and bone regeneration than mineralized dentin. Unlike cell culture therapy, DDM is a matrix-based therapy that includes growth factors. A matrix-based system is a realistic and acceptable treatment, even in developing countries. The aim of this review was to summarize the evidence related to both animal studies and human clinical cases using human dentin materials with a patch of cementum, especially DDM. [ABSTRACT FROM AUTHOR]

The displacement of dental implants into the maxillary sinus is increasingly reported and may lead to serious complications. Better knowledge of this condition could help clinicians improve their practice, but it is difficult to draw conclusions from the current literature. Therefore, a systematic review was performed to describe the main characteristics of dental implant displacement, as well as its management and temporal evolution over a 31-year period. This review was conducted according to the PRISMA methodology. The PubMed/Scopus electronic databases were searched to December 2021. Risk of bias was assessed using the Joanna Briggs Institute tools. A total of 73 articles reporting 321 patients with displaced dental implants were included. Implants located in the upper first molar site were the most frequently involved (23.7%). Displacement occurred mainly during the first 6 months after implant placement (62.6%). The majority became symptomatic (56.2%), most often due to maxillary sinusitis and/or oroantral communication (44.2%). The surgical approaches to remove displaced implants were the lateral approach (38.1%), the Caldwell–Luc approach (27.2%), and endoscopic nasal surgery (23.1%). This review highlights the importance of preventive measures: avoiding implant displacement by careful pre-implantation radiographic analysis, but also preventing infectious complications through early removal of the displaced implant (PROSPERO CRD42021279473). [ABSTRACT FROM AUTHOR]

Determining the emotion of a speaker using their speech utterance by a machine is referred to as Speech Emotion Recognition System. It can greatly enhance the human and machine interaction experience. However, the system faces poor performance due to factors like variations in emotion intensity, speaker, language, and culture. In this study, we have used RAVDESS dataset consisting of speech utterances with two emotional intensities, namely, strong, and normal, raising the recognition difficulty level for the development of an efficient framework for the Speech Emotion Recognition System (SER). For SER, Gender Dependent Training for building the emotion detection models in the Speech Emotion Recognition System is proposed in this research work. The proposed system is less complex and is able to demonstrate good performance using only MFCC features and its variants, namely, delta MFCC and delta-delta MFCC features when compared with the baseline system, which has utilized five different features, namely, MFCC, Mel, Chromogram, Spectral Contrast and Tonnetz. When the average of the speech emotion accuracy of 6 emotions, such as, Sad, Fearful, Calm, Surprised, Disgust and Happy are considered, the proposed system has shown a relatively improved result by 6.90 % over the considered baseline system. [ABSTRACT FROM AUTHOR]

Tolvaptan is an effective drug for the treatment of autosomal dominant polycystic kidney disease, but its use is associated with a significant risk of T-cell-mediated liver injury in a small number of patients. An important clinical conundrum following the contraindication of tolvaptan is whether administration of agents of similar pharmacological action and structure will be tolerated. Herein, we addressed this question through the exposure of tolvaptan-responsive T-cell clones to similar pharmaceutical agents. Whilst lixivaptan and conivaptan did not activate tolvaptan-responsive T-cells, mozavaptan evoked proliferative responses comparable with tolvaptan itself, indicating that there may be collateral immunological intolerance to this compound as a product of sensitization to tolvaptan. [Display omitted] • I n vitro assays show tolvaptan primed T-cells are cross-reactive to the structurally related drug Mozavaptan. • No cross reactivity was seen to the parent compounds lixivaptan or conivaptan for tolvaptan (or metabolite) responsive T-cells. • Data indicates the potential capacity of structurally related compounds to elicit T-cell responses in patients previously sensitised to tolvaptan. • Further work is needed to determine the role of antigen cross-reactivity in the recently observed DILI in lixivaptan treated patients. [ABSTRACT FROM AUTHOR]

The last decade has witnessed rapid advancements in manufacturing technologies for biomedical implants. Additive manufacturing (or 3D printing) has broken down major barriers in the way of producing complex 3D geometries. Electron beam melting (EBM) is one such 3D printing process applicable to metals and alloys. EBM offers build rates up to two orders of magnitude greater than comparable laser-based technologies and a high vacuum environment to prevent accumulation of trace elements. These features make EBM particularly advantageous for materials susceptible to spontaneous oxidation and nitrogen pick-up when exposed to air (e.g., titanium and titanium-based alloys). For skeletal reconstruction(s), anatomical mimickry and integrated macro-porous architecture to facilitate bone ingrowth are undoubtedly the key features of EBM manufactured implants. Using finite element modelling of physiological loading conditions, the design of a prosthesis may be further personalised. This review looks at the many unique clinical applications of EBM in skeletal repair and the ground-breaking innovations in prosthetic rehabilitation. From a simple acetabular cup to the fifth toe, from the hand-wrist complex to the shoulder, and from vertebral replacement to cranio-maxillofacial reconstruction, EBM has experienced it all. While sternocostal reconstructions might be rare, the repair of long bones using EBM manufactured implants is becoming exceedingly frequent. Despite the various merits, several challenges remain yet untackled. Nevertheless, with the capability to produce osseointegrating implants of any conceivable shape/size, and permissive of bone ingrowth and functional loading, EBM can pave the way for numerous fascinating and novel applications in skeletal repair, regeneration, and rehabilitation. Electron beam melting (EBM) offers unparalleled possibilities in producing contaminant-free, complex and intricate geometries from alloys of biomedical interest, including Ti6Al4V and CoCr. We review the diverse range of clinical applications of EBM in skeletal repair, both as mass produced off-the-shelf implants and personalised, patient-specific prostheses. From replacing large volumes of disease-affected bone to complex, multi-material reconstructions, almost every part of the human skeleton has been replaced with an EBM manufactured analog to achieve macroscopic anatomical-mimickry. However, various questions regarding long-term performance of patient-specific implants remain unaddressed. Directions for further development include designing personalised implants and prostheses based on simulated loading conditions and accounting for trabecular bone microstructure with respect to physiological factors such as patient's age and disease status. [Display omitted] [ABSTRACT FROM AUTHOR]

Human dental pulp stem cells (hDPSCs) are multipotent adult stem cells that can differentiate into various lineages such as odontoblasts, osteoblasts, and chondrocytes. Regulation of hDPSCs differentiation with small-molecule compounds can be a useful tool for tissue engineering and regenerative therapy. Forskolin is an agonist of adenylate cyclase that promotes cyclic adenosine monophosphate production. However, the role of Forskolin in regulating the osteogenic differentiation of hDPSCs is still unknown. A cell counting kit-8 (CCK-8) assay was performed to screen out the safety concentrations of Forskolin. Following, quantitative polymerase chain reaction (qPCR) and alizarin red staining were performed to detect bone-related gene expression and mineralized deposit formation. Furthermore, we prepared cell sheets which were followed by a 3D culture for cell pellet formation. Finally, the hDPSC cell pellets were transplanted into immunodeficient mice. CCK-8 assay showed 5 μM and 10 μM Forskolin had no significant inhibition on the proliferation of hDPSCs. The qPCR indicated Forskolin (5, 10 μM) enhanced osteogenic differentiation of hDPSCs by upregulating bone-related genes. Alizarin red staining and its quantification analysis demonstrated Forskolin in 5 μM and 10 μM similarly enhanced the mineralized deposit formation of hDPSCs in vitro. After six weeks of transplantation, immunohistochemical stains showed that osteopontin expression and bone formation were significantly boosted in the Forskolin-treated group than in the normal osteogenic inducing group. Our results indicate Forskolin enhances osteogenic differentiation of hDPSCs in vitro and boosts bone formation in vivo. [ABSTRACT FROM AUTHOR]

• It is unknown whether damage accumulation to ageing, long-lived extracellular matrices is species-dependant or whether it can influence the progression of age-related diseases such as atherosclerosis. • Peptide location fingerprinting, applied to ageing human intervertebral disc (IVD), mouse lung and human arterial atherosclerosis, identified novel target proteins alongside extracellular proteins with structural differences which were conserved between three tissue regions, three organs, two species and sexes. • Alpha-2 macroglobulin and collagen XIV exhibited shared protein structural differences in aged IVD and atherosclerotic artery, providing novel links between an age-related disease and intrinsic ageing. • Crucially, fibronectin, laminins and filamin-A exhibited conserved structural differences between mouse lung and human IVD, providing evidence that ECM networks may be subjected to similar consequences of ageing across both species. Extracellular matrices (ECMs) in the intervertebral disc (IVD), lung and artery are thought to undergo age-dependant accumulation of damage by chronic exposure to mechanisms such as reactive oxygen species, proteases and glycation. It is unknown whether this damage accumulation is species-dependant (via differing lifespans and hence cumulative exposures) or whether it can influence the progression of age-related diseases such as atherosclerosis. Peptide location fingerprinting (PLF) is a new proteomic analysis method, capable of the non-targeted identification of structure-associated changes within proteins. Here we applied PLF to publicly available ageing human IVD (outer annulus fibrosus), ageing mouse lung and human arterial atherosclerosis datasets and bioinformatically identified novel target proteins alongside common age-associated differences within protein structures which were conserved between three ECM-rich organs, two species, three IVD tissue regions, sexes and in an age-related disease. We identify peptide yield differences across protein structures which coincide with biological regions, potentially reflecting the functional consequences of ageing or atherosclerosis for macromolecular assemblies (collagen VI), enzyme/inhibitor activity (alpha-2 macroglobulin), activation states (complement C3) and interaction states (laminins, perlecan, fibronectin, filamin-A, collagen XIV and apolipoprotein-B). Furthermore, we show that alpha-2 macroglobulin and collagen XIV exhibit possible shared structural consequences in IVD ageing and arterial atherosclerosis, providing novel links between an age-related disease and intrinsic ageing. Crucially, we also demonstrate that fibronectin, laminin beta chains and filamin-A all exhibit conserved age-associated structural differences between mouse lung and human IVD, providing evidence that ECM, and their associating proteins, may be subjected to potentially similar mechanisms or consequences of ageing across both species, irrespective of differences in lifespan and tissue function. [ABSTRACT FROM AUTHOR]