Approach to intrapartum fever during the COVID-19 pandemic: A New York City Hospital experience.

The yield of repeat severe acute respiratory syndrome coronovirus 2 (SARS-CoV-2) testing for intrapartum fever in patients admitted to labor and delivery negative for SARS-CoV-2 on admission testing is unknown. From October 2020 to June 2022, we performed a retrospective study of 151/3,168 (4.8%) patients who had repeat testing for intrapartum fever. One (0.7%) patient was SARS-CoV-2-positive suggesting repeat SARS-CoV-2 testing for intrapartum fever is generally not warranted nor is separating birthing dyads while awaiting test results.

In vitro co-expression chromatin assembly and remodeling platform for plant histone variants.

Histone variants play a central role in shaping the chromatin landscape in plants, yet, how their distinct combinations affect nucleosome properties and dynamics is still largely elusive. To address this, we developed a novel chromatin assembly platform for Arabidopsis thaliana, using wheat germ cell-free protein expression. Four canonical histones and five reported histone variants were used to assemble twelve A. thaliana nucleosome combinations. Seven combinations were successfully reconstituted and confirmed by supercoiling and micrococcal nuclease (MNase) assays. The effect of the remodeling function of the CHR11-DDR4 complex on these seven combinations was evaluated based on the nucleosome repeat length and nucleosome spacing index obtained from the MNase ladders. Overall, the current study provides a novel method to elucidate the formation and function of a diverse range of nucleosomes in plants.

Understanding Clinician Knowledge About Race Adjustment in the Vaginal Birth After Cesarean Calculator.

Disparities in maternal health outcomes are striking. Historical and biased clinical support tools have potential to exacerbate inequities. In 2022, NewYork-Presbyterian, with ∼25,000 annual births, and our academic partners, Columbia and Weill Cornell, launched a program to better understand practice patterns and clinician attitudes toward a vaginal birth after cesarean (VBAC) calculator, which predicts VBAC success. This article summarizes the program, focusing on the VBAC calculator utilization survey, which measured provider awareness of the revised calculator and key factors considered in patient counseling. Our preliminary findings warrant future research and education on the calculator's implications for counseling and outcomes.

Altered functional brain networks in problematic smartphone and social media use: resting-state fMRI study.

Nowadays, the limitless availability to the World Wide Web can lead to general Internet misuse and dependence. Currently, smartphone and social media use belong to the most prevalent Internet-related behavioral addiction forms. However, the neurobiological background of these Internet-related behavioral addictions is not sufficiently explored. In this study, these addiction forms were assessed with self-reported questionnaires. Resting-state functional magnetic resonance imaging was acquired for all participants (n = 59, 29 males) to examine functional brain networks. The resting-state networks that were discovered using independent component analysis were analyzed to estimate within network differences. Significant negative associations with social media addiction and smartphone addiction were found in the language network, the lateral visual networks, the auditory network, the sensorimotor network, the executive network and the frontoparietal network. These results suggest that problematic smartphone and social media use are associated with sensory processing and higher cognitive functioning.

Co-Channel Compatibility Analysis of IMT Networks and Digital Terrestrial Television Broadcasting in the Frequency Range 470-694 MHz Based on Monte Carlo Simulation.

According to Resolution 235 adopted during the World Radiocommunication Conference 2015 (WRC-15) and the preliminary agenda item 1.5 of WRC-2023, the International Telecommunication Union Radiocommunication Sector (ITU-R) has been entrusted to carry out sharing and compatibility studies between broadcasting and mobile services in the frequency range 470-694 MHz in Region 1. This study specifically focuses on evaluating electromagnetic compatibility in potential co-channel sharing scenarios between Digital Terrestrial Television Broadcasting (DTTB) and International Mobile Telecommunications (IMT) systems within the 470-694 MHz frequency band that may arise in the foreseeable future. To assess the conditions for coexistence, a Monte Carlo simulation method implemented through the SEAMCAT software version 5.4.2 is employed, examining six potential interference scenarios. The simulation results yield the minimum coordination distance required between IMT and DTTB services in the 470-694 MHz band based on the protection criteria to ensure harmonious coexistence while maintaining satisfactory performance. Furthermore, the study investigates the impact of various factors such as transmitter power, antenna heights, coverage radius, antenna discrimination, and antenna tilt angle on the separation distances. The focus lies primarily on critical interference scenarios across neighboring countries' borders. The simulation outcomes confirm that sharing the same frequency band between IMT and DTTB networks would result in significant mutual interference. Nevertheless, carefully analyzing diverse parameters and assumptions helped provide recommendations to reduce the required separation distances. These findings are valuable for broadcasters, mobile operators, and regulators in establishing the technical coexistence conditions for DTTB and IMT in the new band.

Gray Matter Changes Following Mild COVID-19: An MR Morphometric Study in Healthy Young People.

BACKGROUND: Although COVID-19 is primarily an acute respiratory infection, 5%-40% of patients develop late and prolonged symptoms with frequent neurological complaints, known as long COVID syndrome. The presentation of the disease suggests that COVID infection may cause functional and/or morphological central nervous system alterations, but studies published in the literature report contradictory findings. PURPOSE: To investigate the chronic effects of COVID-19 on cerebral grey matter in a group of young patients without comorbidities, with mild course of COVID infection and no medical complaints at the time of examination. STUDY TYPE: Prospective. POPULATION: Thirty-eight young (age = 26.6 ± 5.0 years; male/female = 14/24), adult participants who recovered from mild COVID infection without a history of clinical long COVID and 37 healthy control subjects (age = 25.9 ± 2.8 years; male/female = 14/23). FIELD STRENGTH/SEQUENCE: Three Tesla, 3D T1-weighted magnetization-prepared rapid gradient-echo, 2D T2-weighted turbo spin-echo. ASSESSMENT: MRI-based morphometry and volumetry along with neuropsychological testing and self-assessed questionnaire. STATISTICAL TESTS: Fisher's exact test, Mann-Whitney U-test, and multiple linear regression analyses were used to assess differences between COVID and healthy control groups. P < 0.05 was used as cutoff for significance. RESULTS: In the COVID group, significantly lower bilateral mean cortical thickness (left/right-hemisphere: 2.51 ± 0.06 mm vs. 2.56 ± 0.07 mm, η2 p = 0.102/2.50 ± 0.06 mm vs. 2.54 ± 0.07 mm, η2 p = 0.101), lower subcortical gray matter (57881 ± 3998 mm3 vs. 60470 ± 5211 mm3 , η2 p = 0.100) and lower right olfactory bulb volume (52.28 ± 13.55 mm3 vs. 60.98 ± 15.8 mm3 , η2 p = 0.078) were found. In patients with moderate to severe anosmia, cortical thickness was significantly lower bilaterally, as compared to patients without olfactory function loss (left/right-hemisphere: 2.50 ± 0.06 mm vs. 2.56 ± 0.05 mm, η2 = 0.173/2.49 ± 0.06 mm vs. 2.55 ± 0.05 mm, η2 = 0.189). Using further exploratory analysis, significantly reduced cortical thickness was detected locally in the right lateral orbitofrontal cortex in the COVID group (2.53 ± 0.10 mm vs. 2.60 ± 0.09 mm, η2 p = 0.112). DATA CONCLUSION: Even without any subjective or objective neurological complaints at the time of the MR scan, subjects in the COVID group showed gray matter alterations in cortical thickness and subcortical gray matter volume. LEVEL OF EVIDENCE: 2 TECHNICAL EFFICACY: Stage 3.

Post-reperfusion acute MR diffusion in stroke is a potential predictor for clinical outcome in rats.

Middle cerebral artery occlusion (MCAO) models show substantial variability in outcome, introducing uncertainties in the evaluation of treatment effects. Early outcome predictors would be essential for prognostic purposes and variability control. We aimed to compare apparent diffusion coefficient (ADC) MRI data obtained during MCAO and shortly after reperfusion for their potentials in acute-phase outcome prediction. Fifty-nine male rats underwent a 45-min MCAO. Outcome was defined in three ways: 21-day survival; 24 h midline-shift and neurological scores. Animals were divided into two groups: rats surviving 21 days after MCAO (survival group, n = 46) and rats dying prematurely (non-survival/NS group, n = 13). At reperfusion, NS group showed considerably larger lesion volume and lower mean ADC of the initial lesion site (p < 0.0001), while during occlusion there were no significant group differences. At reperfusion, each survival animal showed decreased lesion volume and increased mean ADC of the initial lesion site compared to those during occlusion (p < 10-6), while NS group showed a mixed pattern. At reperfusion, lesion volume and mean ADC of the initial lesion site were significantly associated with 24 h midline-shift and neurological scores. Diffusion MRI performed soon after reperfusion has a great impact in early-phase outcome prediction, and it works better than the measurement during occlusion.

Emotional face expression recognition in problematic Internet use and excessive smartphone use: task-based fMRI study.

Growing literature indicates that problematic Internet use (PIU) and excessive smartphone use (ESU) are associated with breakdown of functional brain networks. The effects of PIU&ESU on emotional face expression (EFE) recognition are not well understood, however behavioural investigations and fMRI studies of different addiction forms indicated the impairment of this function. The Facial Emotion Recognition Paradigm was used to probe cortico-limbic responses during EFE recognition. Combined fMRI and psychophysiological analysis were implemented to measure EFE-related functional brain changes in PIU&ESU. Self-reported questionnaires were used to assess PIU&ESU. Positive associations were found between the extent of PIU&ESU and functional connections related to emotional cognitive control and social brain networks. Our findings highlight the involvement of social functioning, especially EFE recognition in PIU&ESU. Therefore, we emphasize that besides the brain's executive and reward systems, the social brain network might be the next candidate to be involved in the pathogenesis of PIU&ESU.

The neural correlates of mental fatigue and reward processing: A task-based fMRI study.

Increasing time spent on the task (i.e., the time-on-task (ToT) effect) often results in mental fatigue. Typical effects of ToT are decreasing levels of task-related motivation and the deterioration of cognitive performance. However, a massive body of research indicates that the detrimental effects can be reversed by extrinsic motivators, for example, providing rewards to fatigued participants. Although several attempts have been made to identify brain areas involved in mental fatigue and related reward processing, the neural correlates are still less understood. In this study, we used the psychomotor vigilance task to induce mental fatigue and blood oxygen-level-dependent functional magnetic resonance imaging to investigate the neural correlates of the ToT effect and the reward effect (i.e., providing extra monetary reward after fatigue induction) in a healthy young sample. Our results were interpreted in a recently proposed neurocognitive framework. The activation of the right middle frontal gyrus, right insula and right anterior cingulate gyrus decreased as fatigue emerged and the cognitive performance dropped. However, after providing an extra reward, the cognitive performance, as well as activation of these areas, increased. Moreover, the activation levels of all of the mentioned areas were negatively associated with reaction times. Our results confirm that the middle frontal gyrus, insula and anterior cingulate cortex play crucial roles in cost-benefit evaluations, a potential background mechanism underlying fatigue, as suggested by the neurocognitive framework.

Language processing in Internet use disorder: Task-based fMRI study.

Internet use disorder (IUD) is generally conceptualized as a fast-growing behavioral addiction. Several structural and functional brain alterations have been revealed in this condition, but previous behavioral studies indicated that language systems may also be impaired. We used a silent word generation task to induce brain activation in Broca's area and other parts of the language system. Blood-oxygen-level-dependent activation analysis and psychophysiological interaction analysis were applied to assess functional brain changes. IUD was measured by the Problematic Internet Use Questionnaire and two additional questions concerning usage time and subjective rating of addiction. According to our key findings, areas strongly related to the default mode network were altered in IUD during the task. Moreover, Broca's area showed altered functional connectivity with other language network and occipital areas in IUD. These findings may address the neural background of decreased verbal fluency performance previously reported in the literature, and we emphasize that beside the brain's reward and inhibitory control systems, the language system is the next candidate to be involved in the pathogenesis of IUD.

Individual differences in the experience of body ownership are related to cortical thickness.

The widely used rubber hand illusion (RHI) paradigm provides insight into how the brain manages conflicting multisensory information regarding bodily self-consciousness. Previous functional neuroimaging studies have revealed that the feeling of body ownership is linked to activity in the premotor cortex, the intraparietal areas, the occipitotemporal cortex, and the insula. The current study investigated whether the individual differences in the sensation of body ownership over a rubber hand, as measured by subjective report and the proprioceptive drift, are associated with structural brain differences in terms of cortical thickness in 67 healthy young adults. We found that individual differences measured by the subjective report of body ownership are associated with the cortical thickness in the somatosensory regions, the temporo-parietal junction, the intraparietal areas, and the occipitotemporal cortex, while the proprioceptive drift is linked to the premotor area and the anterior cingulate cortex. These results are in line with functional neuroimaging studies indicating that these areas are indeed involved in processes such as cognitive-affective perspective taking, visual processing of the body, and the experience of body ownership and bodily awareness. Consequently, these individual differences in the sensation of body ownership are pronounced in both functional and structural differences.

Cerebral Microbleeds May Be Less Detectable by Susceptibility Weighted Imaging MRI From 24 to 72 Hours After Traumatic Brain Injury.

Purpose: A former rodent study showed that cerebral traumatic microbleeds (TMBs) may temporarily become invisible shortly after injury when detected by susceptibility weighted imaging (SWI). The present study aims to validate this phenomenon in human SWI. Methods: In this retrospective study, 46 traumatic brain injury (TBI) patients in various forms of severity were included and willingly complied with our strict selection criteria. Clinical parameters potentially affecting TMB count, Rotterdam and Marshall CT score, Mayo Clinic Classification, contusion number, and total volume were registered. The precise time between trauma and MRI [5 h 19 min to 141 h 54 min, including SWI and fluid-attenuated inversion recovery (FLAIR)] was individually recorded; TMB and FLAIR lesion counts were assessed. Four groups were created based on elapsed time between the trauma and MRI: 0-24, 24-48, 48-72, and >72 h. Kruskal-Wallis, ANOVA, Chi-square, and Fisher's exact tests were used to reveal differences among the groups within clinical and imaging parameters; statistical power was calculated retrospectively for each comparison. Results: The Kruskal-Wallis ANOVA with Conover post hoc analysis showed significant (p = 0.01; 1-ß > 0.9) median TMB number differences in the subacute period: 0-24 h = 4.00 (n = 11); 24-48 h = 1 (n = 14); 48-72 h = 1 (n = 11); and 72 h ≤ 7.5 (n = 10). Neither clinical parameters nor FLAIR lesions depicted significant differences among the groups. Conclusion: Our results demonstrate that TMBs on SWI MRI may temporarily become less detectable at 24-72 h following TBI.

Childhood maltreatment results in altered deactivation of reward processing circuits in depressed patients: A functional magnetic resonance imaging study of a facial emotion recognition task.

IMPORTANCE AND OBJECTIVES: Childhood adversity is a strong risk factor for the development of various psychopathologies including major depressive disorder (MDD). However, not all depressed patients experience early life trauma. Functional magnetic resonance imaging (fMRI) studies using facial emotion processing tasks have documented altered blood-oxygen-level-dependent (BOLD) responses in specific cortico-limbic networks both in MDD patients and in individuals with a history of childhood maltreatment (CM). Therefore, a history of maltreatment may represent a key modulating factor responsible for the altered processing of socio-affective stimuli. To test this hypothesis, we recruited MDD patients with and without of maltreatment history to study the long-term consequences of childhood trauma and examined the impact of CM on brain activity using a facial emotion recognition fMRI task. METHODS: MDD patients with childhood maltreatment (MDD + CM, n = 21), MDD patients without maltreatment (MDD, n = 19), and healthy controls (n = 21) matched for age, sex and intelligence quotient underwent fMRI while performing a block design facial emotion matching task with images portraying negative emotions (fear, anger and sadness). The history of maltreatment was assessed with the 28-item Childhood Trauma Questionnaire. RESULTS: Both MDD and MDD + CM patients displayed impaired accuracy to recognize sad faces. Analysis of brain activity revealed that MDD + CM patients had significantly reduced negative BOLD signals in their right accumbens, subcallosal cortex, and anterior paracingulate gyrus compared to controls. Furthermore, MDD + CM patients had a significantly increased negative BOLD response in their right precentral and postcentral gyri compared to controls. We found little difference between MDD and MDD + CM patients, except that MDD + CM patients had reduced negative BOLD response in their anterior paracingulate gyrus relative to the MDD group. CONCLUSIONS: Our present data provide evidence that depressed patients with a history of maltreatment are impaired in facial emotion recognition and that they display altered functioning of key reward-related fronto-striatal circuits during a facial emotion matching task.

Application of Structural Entropy and Spatial Filling Factor in Colonoscopy Image Classification.

For finding colorectal polyps the standard method relies on the techniques and devices of colonoscopy and the medical expertise of the gastroenterologist. In case of images acquired through colonoscopes the automatic segmentation of the polyps from their environment (i.e., from the bowel wall) is an essential task within computer aided diagnosis system development. As the number of the publicly available polyp images in various databases is still rather limited, it is important to develop metaheuristic methods, such as fuzzy inference methods, along with the deep learning algorithms to improve and validate detection and classification techniques. In the present manuscript firstly a fuzzy rule set is generated and validated. The former process is based on a statistical approach and makes use of histograms of the antecedents. Secondly, a method for selecting relevant antecedent variables is presented. The selection is based on the comparision of the histograms computed from the measured values for the training set. Then the inclusion of the Rényi-entropy-based structural entropy and the spatial filling factor into the set of input variables is proposed and assessed. The beneficial effect of including the mentioned structural entropy of the entropies from the hue and saturation (H and S) colour channels resulted in 65% true positive and 60% true negative rate of the classification for an advantageously selected set of antecedents when working with HSV images.

De novo reconstitution of chromatin using wheat germ cell-free protein synthesis.

DNA is packaged with histones to form chromatin that impinges on all nuclear processes, including transcription, replication and repair, in the eukaryotic nucleus. A complete understanding of these molecular processes requires analysis of chromatin context in vitro. Here, Drosophila four core histones were produced in a native and unmodified form using wheat germ cell-free protein synthesis. In the assembly reaction, four unpurified core histones and three chromatin assembly factors (dNAP-1, dAcf1 and dISWI) were incubated with template DNA. We then assessed stoichiometry with the histones, nucleosome arrays, supercoiling and the ability of the chromatin to serve as a substrate for histone-modifying enzymes. Overall, our method provides a new avenue to produce chromatin that can be useful in a wide range of chromatin research.

Muscle damage in response to a single bout of high intensity concentric exercise in patients with Pompe disease.

BACKGROUND: In Pompe disease, resistance exercise could be an effective treatment to delay motor function impairment, however, the acute effects of this exercise modality are unclear. METHODS: In a prospective cohort study, we compared responses to a single bout of resistance exercise by serum markers of muscle damage and quantitative muscle magnetic resonance imaging (MRI) in patients (n=12) and age- and gender-matched healthy controls (n=12). Participants performed 50 maximal effort concentric knee flexions on a dynamometer. RESULTS: Twenty-four hours after exercise, levels of serum creatine kinase, lactate dehydrogenase and myoglobin increased in controls. In contrast, only myoglobin level increased in patients. All elevated serum markers declined by 48 hours after exercise in both groups. Mild soreness developed at 24 hours, which disappeared at 48 hours in both groups. In controls, MRI R2* relaxation rate reduced immediately and 24 hours after exercise, indicating increased water content and muscle perfusion. In patients, exercise had no effect on R2* values. The resistance exercise did not induce acute strength deficit in patients, rather, patients increased their strength by 24 hours. When serum marker changes were normalized to the magnitude of knee flexor tension developed during exercise, lactate dehydrogenase response was greater in patients. CONCLUSIONS: Late-onset Pompe disease did not exacerbate exercise-induced muscle damage, however, lactate dehydrogenase may be monitored to screen high responders during high intensity resistance exercise interventions.

Microstructural and functional brain abnormalities in multiple sclerosis predicted by osteopontin and neurofilament light.

BACKGROUND: Osteopontin (OPN) is a proinflammatory biomarker, and neurofilament light chain (NFL) levels reflect axonal damage. Resting-state functional MRI (rs-fMRI) defines brain networks during wakeful rest. OBJECTIVE: To examine, if levels of OPN and NFL are associated on the long term with (i) lesion evolution, (ii) changes in normal-appearing white matter (NAWM) microstructure and (iii) functional connectivity in multiple sclerosis (MS). METHODS: Concentration of NFL and OPN in the blood and CSF were related to MRI findings 10.3 ± 2.8 years later in 53 patients with MS. NFL was examined by Simoa method, OPN by ELISA. Lesion volume in the brain and cervical spinal cord was examined by 3D FLAIR images. Voxel-wise images of fractional anisotropy (FA), axial diffusivity (AD), mean diffusivity (MD), and radial diffusivity (RD) were examined by tract-based spatial statistics corrected for gender, age and lesion volume. Metabolites were examined by single-voxel MR-spectroscopy in the NAWM. Fifty-five default mode network connections were examined by rs-fMRI corrected for gender, age, MS subtype and current therapy as covariates. RESULTS: While NFL in paired serum and CSF positively correlated (p = 0.019), there was no correlation between serum and CSF OPN. Higher OPN levels in the CSF but not in the serum showed association with increased brain WM lesion volume (p = 0.009) in 10.3 ± 2.8 years. Higher OPN in the CSF was associated with reduced FA, increased MD, and reduced RD in different NAWM areas 10.3 ± 2.8 years later. Higher OPN in the serum and CSF were associated with increased connectivity strength between the medial prefrontal cortex (MPFC) and other regions except with inferior parietal lobule. NFL in the CSF and in the serum was associated with decreased connectivity strength except for ventral MPFC-hippocampal formation. Neither serum OPN nor NFL at the time of the MRI were associated with functional connectivity changes. CONCLUSION: While serum NFL levels reflects CNS production, OPN in serum and CSF may have different cellular sources. OPN within the CSF but not in the serum may forecast development of lesions and microstructural abnormalities in 10 years, indicating the detrimental role of CNS inflammation on the long-term. Although both OPN and NFL in the CSF were associated with functional connectivity changes in 10 years, NFL was associated with decreased strength possibly indicating general axonal loss. In contrast, the positive association of OPN levels in the CSF with increased connectivity strength in 10 years may point to adaptive re-organization due to inflammatory WM lesions and microstructural changes.

Provocation and prediction of visual peripersonal neglect-like symptoms in preoperative planning and during awake brain surgery.

Neglect is a severe neuropsychological/neurological deficit that usually develops due to lesions of the posterior inferior parietal area of the right hemisphere and is characterized by a lack of attention to the left side. Our case is a proven right-handed, 30-year-old female patient with a low-grade glioma, which was located in the temporo-opercular region and also in the superior temporal gyrus of the right hemisphere. Upon presurgical planning, the motor, language, and visuospatial functions were mapped. In order to achieve this, the protocol for routine magnetic resonance imaging and navigated transcranial magnetic stimulation has been expanded, accordingly.

Fast 3 T nigral hyperintensity magnetic resonance imaging in Parkinson's disease.

The absence of nigral hyperintensity is a promising MR marker for Parkinson's disease (PD), but its small size imposes limitations on its routine use. Our aim was to compare Multi Echo Data Image Combination (MEDIC), segmented echo-planar imaging (EPISEG) and fluid-attenuated inversion recovery (FLAIR) sequences, as well as both magnitude (MAG) and susceptibility-weighted imaging (SWI) reconstructions of single-echo gradient echo for nigral hyperintensity imaging. Twenty-five healthy and twenty PD subjects were included. Sensitivity to motion artefacts, confidence of the radiologist in interpretation, rate of nondiagnostic scans and diagnostic accuracy were assessed. EPISEG was less motion-sensitive than MEDIC, MAG, and SWI, while FLAIR was less motion-sensitive than MAG and SWI. The reviewers were more confident when using EPISEG compared to any other techniques and MEDIC was superior to FLAIR. The proportions of nondiagnostic scans were lower for EPISEG than for other sequences. The best diagnostic performance was achieved for EPISEG (sensitivity = 65%, specificity = 96%). Using EPISEG, the absence of nigral hyperintensity in PD was associated with higher Hoehn-Yahr stage and MDS-UPDRS II + III. Nigral hyperintensity may be intact at the very early stages of PD. The promising properties of EPISEG may help the transfer of nigral hyperintensity imaging into daily clinical practice.

Reconstitution of Drosophila and human chromatins by wheat germ cell-free co-expression system.

BACKGROUND: Elaboration of the epigenetic regulation of chromatin is a long-standing aim in molecular and cellular biology. Hence, there is a great demand for the development of in vitro methods to reconstitute chromatin that can be used directly for biochemical assays. The widely used wheat germ cell-free protein expression method provides broad applications to investigate the function and structure of eukaryotic proteins. Such advantages, including high translation efficiency, flexibility, and possible automatization, are beneficial for achieving native-like chromatin substrates for in vitro studies. RESULTS: We describe a novel, single-step in vitro chromatin assembly method by using the wheat germ cell-free protein synthesis. We demonstrated that both Drosophila and human chromatins can be reconstituted in the course of the in vitro translation of core histones by the addition of chromatin assembly factors, circular plasmid, and topoisomerase I in an ATP-dependent manner. Drosophila chromatin assembly was performed in 4 h at 26 °C, in the presence of premixed mRNAs encoding the core histones, dAcf1/dISWI chromatin remodeling complex, and nucleosome assembly protein, dNAP1. Similarly, the human chromatin was assembled by co-expressing the human core histones with Drosophila chromatin remodeling factor, dISWI, and chromatin chaperone, dNLP, for 6 h at 26 °C. The presence of reconstituted chromatin was monitored by DNA supercoiling assay, also the regular spacing of nucleosomes was assessed by Micrococcal nuclease assay. Furthermore, Drosophila linker histone H1-containing chromatin was reconstituted, affirming that the in vitro assembled chromatin is suitable for downstream applications. CONCLUSIONS: The method described in this study allows the assembly of Drosophila and human chromatins, possibly in native-like form, by using a wheat germ cell-free protein expression. Although both chromatins were reconstituted successfully, there were unexpected differences with respect to the required ratio of histone-coding mRNAs and the reaction time. Overall, our new in vitro chromatin reconstitution method will aid to characterize the unrevealed structure, function, and regulation of chromatin dynamics.