Bioelectrical bioimpedance analysis was used for body composition measurements. Resting metabolic equivalent (1-MET) was 3.31 ± 0.66 mL·kg−1·min−1. Participants completed an incremental walking test with oxygen uptake measurements to individualize the treadmill walking speed at 5-MET. In a randomized, double-blind, cross-over design, the 30 min morning walks were in the same phase of each participant’s menstrual cycle. No changes by NZBC extract were observed for walking-induced heart rate, minute ventilation, oxygen uptake, and carbon dioxide production. NZBC extract enhanced fat oxidation . There was a significant correlation for changes in fat oxidation with body mass index; body fat% in legs, arms, and trunk; and a trend with fat oxidation at rest but not with body mass and habitual anthocyanin intake. The NZBC extract responsiveness of walking-induced fat oxidation is body composition-dependent and higher in young-adult females with higher body fat% in legs, arms, and trunk.New Zealand blackcurrant (NZBC) extract enhanced cycling-induced fat oxidation in female endurance athletes. We examined in recreationally active females the effects of NZBC extract on physiological and metabolic responses by moderate-intensity walking and the relationship of fat oxidation changes with focus on body composition parameters. Twelve females (age: 21 ± 2 y, BMI: 23.6 ± 3.1 kg·m The energy requirements of moderate-intensity exercise are provided primarily by the oxidation of carbohydrates and lipids . The reln = 12, age: 21 ± 2 years; height: 166 ± 7 cm; body mass: 65 ± 11 kg; BMI: 23.7 ± 3.1 kg·m−2) recruited for the study were healthy, recreationally active Caucasian university students and staff. All females had a regular menstrual period, were non-smokers, and were not taking other dietary supplements. Participants provided written informed consent, and health status was confirmed by a health history questionnaire. The physical activity level of the female participant’s physical activity level was quantified with the short version of the International Physical Activity Questionnaire (2749 ± 2106 MET·week−1) [The female participants (·week−1) . The pre−1; range: 5 to 88 mg·day−1).The study had a randomized, placebo-controlled, cross-over design. Female participants visited the laboratory for one preliminary and two experimental sessions. For each session, participants abstained from unaccustomed and intense exercise for 48 h, had no alcohol and caffeine intake for 24 h, and were at least 2 h postprandial after consuming a breakfast of one slice of bread and a glass of water . Session−1·min−1). Heart rate at rest was 69 ± 9 beats·min−1. Blood pressure at rest was taken twice  and averaged . Subsequently, participants completed an incremental-intensity walking protocol on a treadmill . Treadmill incline was 1%. Participants completed 5 × 8-min stages starting at 2 km∙h−1, progressing by 1 km∙h−1 until a speed of 6 km∙h−1 was reached [2 = 0.9626 ± 0.0259). The linear relationship between walking speed and metabolic equivalent allowed for each participant to establish the walking speed at 5-METs .In the preliminary session, height  and body mass  were measured as well as body composition . Subsequently, participants were fitted with a heart-rate monitor  Ltd., Warwick, UK). Participants then rested for 30 min in a chair, followed by 2 × 10 min expired air collection using Douglas bags  to determine the oxygen consumption at rest. The lowest value for oxygen consumption at rest was taken as the one metabolic equivalent (1-MET)   or placebo pills  were taken for 7 days. The final two capsules were taken 2 h before visiting the laboratory for the experimental sessions. Participants were allowed a breakfast consisting of one slice of bread and a glass of water 3 h before visiting for the experimental sessions.For the experimental sessions, two capsules of NZBC extract  at the speed established in the preliminary session with recording of heart rate  Ltd., Warwick, UK) and expired-air collection from 7–10, 17–20, and 27–30 min. Rates of whole-body fat and carbohydrate oxidation were calculated with equations below from Jeukendrup and Wallis  and the The respiratory exchange ratio was calculated by dividing the volume of carbon dioxide produced by the volume of oxygen consumed.t-tests were used to compare all the parameters between the placebo and NZBC extract conditions. Values are reported as mean ± SD and 95% confidence intervals. Statistical significance was accepted at p < 0.05. p-Values of 0.05 ≥ p ≤ 0.1 were interpreted according to guidelines by Curran-Everett and Benos [d < 0.2), small (d = 0.2–0.49), moderate (d = 0.5–0.79), and large (d ≥ 0.8). Pearson correlation coefficients were calculated and tested for significance for the relationships between habitual anthocyanin intake (not considering the NZBC extract intake); fat oxidation at rest; body mass; body mass index; fat% of the legs, arms, and trunk; and changes of walking-induced fat oxidation with intake of NZBC extract in comparison to walking-induced fat oxidation in the placebo condition. The ∆ FAO is the fat oxidation with intake of New Zealand blackcurrant extract minus the fat oxidation with intake of placebo. The required sample size was not calculated, and the number of 12 participants was lower than previous studies with observations of an effect of NZBC extract on exercise-induced fat oxidation . Physiological and metabolic responses by the moderate-intensity walk were measured from 7–10, 17–20, and 27–30 min during the walk and averaged. All parameters were tested for normality with the D’Agostino and Pearson omnibus test. Two-tailed paired sample nd Benos . Cohen’s [n = 15 ; females [n = 16 ).−1, 95% CI ; NZBC extract: 120 ± 14 beats·min−1, 95% CI ; p = 0.58, d = −0.06), minute ventilation ; NZBC extract: 24.4 ± 6.3 L·min−1, 95% CI , p = 0.14, d = −0.20), oxygen uptake ; NZBC extract: 15.8 ± 1.8 mL·kg−1·min−1, 95% CI , p = 0.45, d = −0.18), and carbon dioxide production ; NZBC extract: 13.1 ± 1.4 mL·kg−1·min−1, 95% CI , p = 0.17, d = −0.40). The absence of an effect on the physiological responses by NZBC extract indicate no change in the regulatory mechanism for exercise-induced heart rate and respiratory demands.During the moderate-intensity treadmill walk, the NZBC extract had no effect on heart rate  (−1), NZBC: 95% CI , p = 0.03, d = −0.56) (−1), NZBC extract: 95% CI , p = 0.005, d = 0.59) , NZBC extract: 95% CI , = −0.69) A, 10.8% = −0.56) B, and 25 = 0.59) C. The 102 = 0.04, p = 0.56)   A. The ha = 0.09) B. This s2 = 0.23, p = 0.11)      A. Howeve= 0.008) B, body f= 0.005) C, body f= 0.016) D, and bo= 0.019) E and theThe observations on body composition parameters suggest an enhanced response for exercise-induced fat oxidation with intake of NZBC extract for females with higher body fat in the legs, arms, and trunk. The significant correlations for the relationship between body mass index; body fat% in the legs, arms, and trunk; and changes in walking-induced fat oxidation may suggest differences in adipocyte sensitivity in response to intake of NZBC extract in females with higher body fat%.2max [The present study presents novel findings on the effect of intake of anthocyanin-rich New Zealand blackcurrant extract on the physiological and metabolic responses during moderate-intensity walking in recreationally active females. Recently, Elliott-Sale et al.  emphasiz V˙O2max . In the  V˙O2max , no info V˙O2max   that resting fat oxidation correlated (R = 0.55) with exercise-induced maximal fat oxidation. In addition, in the same study, 2max correlated (R = 0.44) with resting fat oxidation [2max and thus baseline cardiovascular fitness may be predictive of the response to enhanced walking-induced fat oxidation with intake of New Zealand blackcurrant extract.Robinson et al.  reportedxidation . In the The present study used a convenience sample of recreationally active female participants as the primary aim was to examine the effect of intake of New Zealand blackcurrant extract on walking-induced fat oxidation. In a previous study from our group, Matcha green tea drinks in females enhanced walking-induced fat oxidation by 18% . It is pA limitation of the present study was that participants did not record a 48 h food diary for the two visits with taking placebo or New Zealand blackcurrant extract. However, although previous studies ,26 had dIn conclusion, in recreationally active adult females, 7-day intake of NZBC extract substantially enhanced fat oxidation by 30 min of moderate-intensity walking exercise. The enhanced walking-induced fat oxidation by intake of NZBC extract in recreationally active females was body composition-dependent.


The findings show that accelerated spiking during NMDA and glutamate is associated with enhanced or attenuated LC synchrony, respectively, causing distinct LFP pattern transformations. Shaping of LC population discharge dynamics by ionotropic glutamate receptors potentially fine-tunes its influence on brain functions.Locus coeruleus (LC) neurons are controlled by glutamatergic inputs. Here, we studied in brain slices of neonatal rats NMDA and glutamate effects on phase-locked LC neuron spiking at ~1 Hz summating to ~0.2 s-lasting bell-shaped local field potential (LFP). NMDA: 10 μM accelerated LFP 1.7-fold, whereas 25 and 50 μM, respectively, increased its rate 3.2- and 4.6-fold while merging discrete events into 43 and 56% shorter oscillations. After 4–6 min, LFP oscillations stopped every 6 s for 1 s, resulting in ‘oscillation trains’. A dose of 32 μM depolarized neurons by 8.4 mV to cause 7.2-fold accelerated spiking at reduced jitter and enhanced synchrony with the LFP, as evident from cross-correlation. Glutamate: 25–50 μM made rhythm more irregular and the LFP pattern could transform into 2.7-fold longer-lasting multipeak discharge. In 100 μM, LFP amplitude and duration declined. In 25–50 μM, neurons depolarized by 5 mV to cause 3.7-fold acceleration of spiking that was less synchronized with LFP. Both agents: evoked ‘post-agonist depression’ of LFP that correlated with the amplitude and kinetics of V Note that the IR score has no unit. The lower the value, the more regular the network rhythm.p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001) was assessed by a two-tailed paired t test and one-way ANOVA with Dunnett’s post-test  using Prism software .Values are given as means ± SD and n-values correspond to measurements in one slice per animal with a total of 58 rats used. Significance  (In >70% of slices (n = 29), the rate, duration and amplitude of LFP were regular A–C. In fe: 0.98) E.The LFP could show an irregularity regarding rate, duration and amplitude not only in distinct slices, but also between cycles in the same slice . This isThe above findings indicate that the LFP envelope is bell-shaped due to a normal distribution of the occurrence of single spikes that jitter similarly around the LFP peak. Next, dose–response relationships were studied for the effects of NMDA and glutamate to determine the thresholds for putative LFP pattern transformation and elucidate whether high concentrations of the agents perturb network rhythm or cellular spiking as previously reported .NMDA effects on LFP: Examples are given in In the four slices, LFP duration decreased from 183 ± 38 to 133 ± 18 ms at 10 μM, 105 ± 19 early at 25 μM, 78 ± 23 ‘late’  at 25 μM and 81 ± 5 ‘early’ at 50 μM A,B. ConvAmplitude fluctuations of early LFP oscillations tended to increase from <15% at 25 μM to 40–50% at 50 μM. When LFP oscillation trains at 50 μM (n = 4) were stable in their early phase for 3 cycles, their duration was 6.06 ± 2.64 s and rhythmic LFP blockade lasted 0.94 ± 0.34 s.p = 0.002) , whereas the LFP rate showed a trend to rise from 42.3 ± 7.2/min to 57.3 ± 15.6 events/min (p = 0.13).Glutamate-effects on LFP: An example is given in The above described NMDA-evoked shortening of LFP event duration indicates an increase in network synchrony, whereas increased LFP duration and irregularity indicates the opposite for glutamate. To substantiate this assumption, cross-correlation analysis was performed between the LFP peak and intracellular spike occurrence.m from its ‘resting’ value of −45 mV. This depolarization accelerated spontaneous discharge in this neuron from 48 to 228 spikes/min. The correlogram for the recording in m, 5.0 ± 1.1 mV (p < 0.001) for the glutamate depolarization and 52.1 ± 4.6 vs. 194 ± 61.0 spikes/min (p < 0.001)  for control vs. glutamate.Glutamate-evoked attenuated synchrony: Cross-correlation analysis of glutamate effects was done in six slices. In three cases, 25 μM transformed the LFP pattern within 2–3 min. In the other three cases, the glutamate dose was increased after 5 min, from 25 μM to 50 μM, which transformed the pattern after a further 2–3 min. An example for the latter approach is shown in m, 8.4 ± 2.3 mV (p < 0.001) for the NMDA depolarization and 49.4 ± 6.1 vs. 355 ± 35.6 spikes/min (p < 0.001)  for control vs. NMDA.NMDA-evoked enhanced synchrony: Based on the above results from the dose–response relationships, 32 μM NMDA was applied to seven neurons for cross-correlation analysis, as this dose should rapidly transform the LFP pattern into oscillations while not immediately evoking oscillation trains. As exemplified in In line with the assumption based on these findings that network synchrony was enhanced, the jitter of the time point of spiking in these neurons related to the LFP decreased from 103.8 ± 34.4 ms in control to 13.9 ± 2.25 ms in NMDA  D.Regularity of rhythm: A further measure of rhythmic neural network behavior is the irregularity score . For them. A total of 32 μM NMDA was used again, as oscillation trains in 25 μM tended to start only after several minutes, whereas 50 μM depressed them too quickly. m repolarized for ~0.5 s close to its resting value of −48 mV with concomitant spiking arrest. Then, a crescendo-like ‘ramp’ depolarization developed that caused spike onset after ~0.5 s, when LFP oscillations also recurred  and spike rate during the last second was 462 ± 45.7 spikes/min compared to 49.3 ± 6.7 spikes/min in control (p < 0.001) (thus 9.3-fold faster). Vm repolarized after the end of the train to −46.8 ± 2.9 mV (p = 0.06) and the pause until the next train started was 1.0 ± 0.1 s.For six neurons, several parameters were analyzed for the most stable and longest oscillation train early during their appearance. Specifically, the train lasted for 7.1 ± 0.9 s, NMDA caused at the end of the train a 20.0 ± 6.3 mV depolarization from a resting Vm hyperpolarized, and, at the same time, cellular spiking and LFP were blocked.Apart from their opposite action on LC neuron synchrony, both agents exerted a similar inhibitory effect. Specifically, within 0.5–2 min following the start of washout of NMDA A or glutm at the start of such PAD from a resting value −48.7 ± 2.5 mV by 22.5 ± 6.4 mV (p = 0.006). Vm hyperpolarized during the inhibition maximally to −65.7 ± 5.0 mV (p = 0.005) and the effect was reversed after 9.0 ± 1.4 min. In four different neurons, glutamate had depolarized Vm at the start of PAD from −48.5 ± 3 mV by 5.2 ± 3.2 mV (p = 0.04), the maximal hyperpolarization was to −52.0 ± 1.6 mV (p = 0.03) and the effect was reversed after 1.5 ± 0.4 min.In four neurons, NMDA had depolarized VNMDA binds selectively to NMDA-type iGluR, whereas glutamate can act on NMDA- and AMPA/Kainate-type iGluR  plus divm started to repolarize. Intracellular spikes and LFP recurred after 80 s and control-like LFPs and spike rates were seen in the presence of glutamate and kynurenic acid after 3 min (m at the start of kynurenic acid-evoked hyperpolarization from a resting Vm of −44.7 ± 0.5 mV by 3.5 ± 0.5 mV (p = 0.001), the maximal hyperpolarization was to −52.5 ± 2.8 mV (p = 0.009) and the effect reversed after 2.3 ± 0.4 min.In the neuron of er 3 min A. SubseqThe late countering kynurenic acid effect shows that glutamate effects are mediated by iGluR. To exclude a novel type of inhibitory effect of kynurenic acid in the initial phase of application, 2.5 mM of the agent was added to control superfusate in 4 slices with regular LFP pattern. Kynurenic acid had no effect B as in oNMDA- and glutamate-related blockade of intracellular spiking correlated well with inactivation of the majority of the 3–10 neurons comprising the LFP. However, early during kynurenic acid application in glutamate small amplitude random LFP spiking was seen, whereas normal LFP events and neuronal spikes were blocked A. SimilaPerspective article in this Special Issue [2+-dependent ion conductances causing rhythmic depolarization of gap junction-coupled neonatal LC neurons. However, as also noted in the latter report, phasic afferent activity in the slice model can also be studied, namely with tetanic stimulation of slice areas outside the LC soma region. The present study is the first demonstration of LC neural network discharge pattern transformation by iGluR. We also found that NMDA and glutamate evoke PAD. Possible mechanisms and consequences for LC network organization are discussed in the following.We found that NMDA accelerates the LFP rhythm and enhances the synchrony of LC neuron spiking, whereas glutamate makes the rhythm less regular and weakens synchrony. In a al Issue , we refe+ or during recovery from opioid inhibition [+ nor opioids change LC excitability to cause release of glutamate, which might then act on oscillation-generating NMDA receptors . Glutamate elicited LFP pattern transformation at 25–50 μM. This can be functionally relevant in the intact neonatal LC as extracellular glutamate rises in vivo from a 1–10 μM baseline to >20 μM like during treatment with μ-opioid (ant)agonists [In our previous study, we used suction electrode recording to unravel that LC neurons show jittered spiking that is yet phase-locked to generate rhythmic ~0.2-lasting LFP discharge . Here, whibition . Here, gagonists ,39.The finding that kynurenic acid reversed the glutamate-evoked LFP pattern transformation shows lack of involvement of metabotropic glutamate receptors. Contrary to our observation of distinct LFP network discharge pattern transformations, single neuron recording here and previously ,16,17,40A plus glycine receptor blockades [Perspective article in this issue [2+/calmodulin-dependent protein-kinase-I activation, which enhances weak coupling of non-neighboring neurons [m close to STO peaks, thus altering spike threshold [NMDA-evoked LFP oscillations were faster, shorter and more regular than control events, whereas glutamate made the rhythm apparently more irregular and more than doubled the duration of separate events if present. The assumption that NMDA enhances and glutamate and weakens neonatal LC synchrony was confirmed by cross-correlating neuronal spiking with LFP events . Moreovelockades , similarlockades ,41,42. Tlockades ,44,45,46is issue , NMDA st neurons . Contrarhreshold . Thus, g+-dependent K+ current causes PAD after glutamate, but not NMDA [2+-activated K+ channels are involved as ongoing neuronal depolarization likely leads to a cytosolic Ca2+ rise mainly mediated by voltage-activated Ca2+ channels [2+ buffering. Moreover, the pacemaker rate in LC neurons of mouse slices is regulated by cooperation of L- and T-type Ca2+ with SK2 channels [2+ rises reach SK channel activation threshold. Our finding that NMDA-evoked neuronal rhythmic depolarization and spiking persists during current-evoked hyperpolarization indicates that these events are voltage-clamped by gap junction-coupled neighboring neurons as in the juvenile rat CA3 hippocampus [m oscillations [Whereas LFP pattern transformation was associated with increased spiking, NMDA and glutamate also acted in an inhibitory way. Specifically, early during washout of both agents, neuronal hyperpolarization was associated with PAD of LFP, whereas periodic inhibition of LFP oscillations developed later during NMDA application. In vivo, PAD occurs after sensory stimulation-evoked discharge ,49. Reganot NMDA . As bothchannels . In factchannels  and slicchannels , currentchannels ,54. Enhapocampus . Pharmacllations ,25,26,27Perspective article in this Special Issue [1 receptor, they may release lactate to excite in positive feedback LC neurons via a novel receptor [Perspective article in this issue [We hypothesized above that opposite iGluR effects on LFP patterns may be caused by differences in neuronal properties. Indeed, ventrally-located adult rat LC neurons with shorter spikes and smaller afterhyperpolarizations than LC core neurons act as a ‘pontospinal-projecting module’ , whereasal Issue . Two finreceptor . Our batis issue .+ and opioids [Our finding that already complex LC population discharge can transform into different LFP patterns during NMDA and glutamate (here) or during high K opioids  indicate opioids . The aut opioids ,9,10,11  opioids . A relat opioids . Moreove opioids . These i opioids ,61,62,63 opioids .


With the help of the complementary advantages of different explosophoric groups and diverse weak interactions, BITE-101 is superior to the benchmark explosive HMX in all respects, including higher density of 1.957 g·cm−3, highest decomposition temperature of 295 °C (onset) among CHON-based high explosives to date and superior detonation velocity and pressure , impact and friction sensitivities , thereby showing great potential for practical application as replacement for HMX, the most powerful military explosive in current use.The design and synthesis of novel energetic compounds with integrated properties of high density, high energy, good thermal stability and sensitivities is particularly challenging due to the inherent contradiction between energy and safety for energetic compounds. In this study, a novel structure of 4-amino-7,8-dinitropyrazolo-[5,1- The design of high energy density materials (HEDM) with good detonation performance but which are also safe to handle is challenging. Here, the authors synthesize a PTX analogue and incorporate explosophoric groups to obtain an HEDM with improved detonation performance but low impact and friction sensitivity. Energetic compounds, which are the major energy components in HEDMs, are structurally composed of backbones and explosophoric groups3. The explosophoric groups such as nitramine, azido, nitro, amino, N-O, nitroform etc. are the most commonly selected groups to increase the energy of energetic compounds. In terms of practical applications, a restricted process is required to examine their properties such as thermal stability, sensitivities, detonation velocity and pressure, etc., the worst performance of an energetic compound determines if it holds the application prospects5. The utilization of multi kinds of explosophoric groups to manipulate substitution positions of energetic compounds to regulate the properties of energetic compounds is a commonly used method. Octogen (HMX) is the most powerful military explosive in current use7, it has a density of 1.905 g cm−3, an onset decomposition temperature of 279 °C, detonation velocity of 9144 m s−1, and detonation pressure of 39.2 GPa. During the last few decades, lots of efforts have been devoted to synthesize CHON-based high explosives16. As shown in Fig. −1 were collected and summarized. However, most of those high explosives decompose below 200 °C, which is difficult to meet the requirement for practical applications4. Furthermore, regards to the decomposition temperature, only 2,4,6-triamino-5-nitropyrimidine-1,3-dioxide (ICM-102)17 (Td = 284 °C) was reported to have a higher decomposition compared to that of HMX to date. The search for new energetic compounds with superior performances that outperforms HMX remains a long-term challenge.High energy density materials (HEDMs) present a class of materials that possess high densities and high detonation properties, since the existing contradiction between energy and safety, the pursuing of energetic compounds with good comprehensive properties is a desirable goal for researchers19. The incorporation of a nitro group or N-O group with an amino group to create novel energetic compounds with high density, excellent thermal stability as well as low sensitivity is a well-recognized strategy26. A representative fused energetic compound 3,7,8-trinitropyrazolo  triazin-4-amine (PTX)24, which contains pyrazole-triazine fused ring and two kinds of explosophoric groups exhibit a high density of 1.946 g cm−3, the high decomposition temperature of 246 °C and comparable detonation properties to that of RDX7. Furthermore, the existing explosophoric groups of amino and nitro in PTX offers more possibility form hydrogen bonding interaction, therefore, giving better impact and friction sensitivity  than those of RDX  in which the nitramine is only explosophoric group in the structure. Maintaining the diversity of explosophoric groups in backbones is beneficial to achieve balanced comprehensive properties for energetic compounds.The construction of fused ring structures is an effective way to enhance the thermal stability of energetic compounds2 of PTX with an N-O moiety, we envisioned that three different explosophoric groups could lead to diverse weak interactions while maintaining the zero-oxygen balance (CO as the product) and increasing the varieties of explosophoric group and also the heat of formation compared to that of PTX, hence make the newly designed compound with better detonation performances and higher stabilities than PTX. Herein, we present the synthesis of a novel fused ring energetic compound 4-amino-7,8-dinitropyrazolo- -tetrazine 2-oxide (named BITE−101), it was readily prepared by a three-step reaction. The structures and properties of prepared compounds were characterized by multinuclear NMR, infrared spectroscopy, elemental analysis, and differential scanning calorimetry (DSC). As expected, the more nitrogen atoms in the fused backbone and more kinds of explosophoric groups compared to those in PTX, affording BITE-101 a high density of 1.957 g cm−3, highest decomposition temperature  among those CHON-based explosives with D > 9000 m s−1  with a yield of 86%  and unit cell size V = 828.91(8) Å with a crystal density of 1.957 g cm−3 (298 K). The C-N bond length in the pyrazole fragment lies between 1.435–1.467 Å, which is similar to those in PTX (1.436–1.458 Å)24.The structure of BITE-101 was further determined by an X-ray single-crystal diffraction study of the crystals grown from methanol solution at room temperature. As shown in Fig. 2 is 1.308 Å, the N-N bond length ranges from 1.328 to 1.356 Å, and the N-O bond length (1.240 Å) is shorter than N-O bond length in ICM-10217 (1.253(5) Å) and PTX (1.274(8) Å). In the packing diagram of BITE-101, four independent molecules in different orientations form a wave-like stacking structure, the interlayer spacing between adjacent parallel molecules of BITE-101 is 3.165 Å, which is shorter than that in PTX (3.450 Å) , the shorter average hydrogen bond length was also observed  Fig. . The two−1 to date. To achieve a deeper understanding of the superior thermal stability of BITE-101, the Independent Gradient Model based on Hirshfeld partition (IGMH)30 and bond dissociation energy (BDE) calculations were performed. As shown in Fig. 31 for BITE-101 was calculated by comparing to those of PTX was shown in Fig. 2: 249.423 kJ mol−1) is higher than that in PTX (C5-NO2: 247.385 kJ mol−1), demonstrating better thermal stability for BITE-101, which agrees well with the experimental results.BITE-101 decomposes at 295 °C (onset) with a peak temperature at 300 °C  is significantly better than that of HMX (IS = 7.4 J).The heat of formation (HOF) of BITE-101 was calculated by using Gaussian 09 program2, C-NH2, N-O) in the structure of BITE-101 help to form extensive hydrogen bonding and π-π interactions and result in BITE-101 featuring a high measured density of 1.957 g cm−3, the highest decomposition temperature of 295 °C (onset) among CHON-based high explosives (D > 9000 m s−1) to date, better impact sensitivity (18 J) and friction sensitivity (128 N) along with a superior detonation velocity (9316 m s−1) to the benchmark explosive HMX, demonstrating great application prospects as a new generation of high energy density materials. The investigation of the application for BITE-101 is currently in progress in our lab.In summary, a high energy density compound BITE-101 was synthesized through a novel cyclization strategy in three steps without involving diazo, azide highly sensitive, or other toxic intermediates. The tri- explosophoric groups  spectra were recorded on an FT-IR spectrometer (Thermo Nicolet AVATAR 370). Decomposition (onset) temperature were recorded on a differential scanning calorimeter  at a scan rate of 5 °C min−1. Elemental analyses  were performed on a Vario Micro cube Elementar Analyzer. Impact and friction sensitivity measurements were made using a standard BAM Fallhammer and a BAM friction tester.All reagents were purchased from Energy Chemical or Aladdin in analytical grade. DANP was prepared from the literature’s methodf = 0.55) detects that the reaction is completed. The precipitate was filtered off, washed with water, and dried in the air. The crude brownish-yellow solid of compound 1 was obtained . Td (onset) 237 °C. 1H NMR  δ 9.09 , 7.55, 5.90. 13C NMR  δ 155.4, 150.9, 148.5, 109.1 ppm. IR (KBr) ν̃ = 3477, 3553, 3234, 1646, 1611, 1568, 1519, 1490, 1353, 1253, 1203, 1168, 1137, 1047, 954, 820, 798, 770, 690, 647, 612, 566, 543, 409 cm−1. Elemental analysis (%) for C4H6N8O4 (230.05) Calcd: C 20.88, H 2.63, N 48.69. Found: C 20.83, H 2.39, N 49.02.1-Methyl-2-nitro-1-nitrosoguanidine  was added to a solution of 3,5-diamino-4-nitro-pyrazole  in 30 ml of ethanol, and the mixture was heated to 80 °C. After refluxing for 12 h, TLC  was added to the mixed solution at 0 °C in batches after the addition was completed. The reaction temperature gradually rose to room temperature and reacted for another 3 h. After the reaction was completed, which was monitored by TLC , the reaction solution was poured into ice water, and a yellow solid is precipitated. The precipitate was collected by filtration and washed with water to give 2 . Td (onset) 204 °C. 1H NMR  δ 9.88 , 8.82 . 13C NMR  δ 155.1, 150.2, 148.5, 109.0 ppm. IR (KBr) ν̃ =3439, 3374, 3304, 3248, 1655, 1565, 1530, 1504, 1451, 1434, 1377, 1346, 1313, 1255, 1214, 1172, 1085, 1034, 948, 879, 820, 783, 773, 762, 756, 715, 683, 674, 624, 517, 462 cm−1. Elemental analysis (%) for C4H4N8O6 (260.03) Calcd: C 18.47, H 1.55, N 43.08. Found: C 18.16, H 1.89, N 42.71.About 10 mL of 30% H3 cooled to 0 °C was added 100 mg Compound 2  in batches, after then, the reaction mixture was slowly warmed up to room temperature and stirred for another 3 h. After the reaction was completed it was monitored by TLC . Then, the reaction mixture was poured into ice water to precipitate a yellow solid. The solid was collected by filtration and washed with water to obtain a light yellow solid powder of BITE-101 . Td (onset) 295 °C. 1H NMR  δ 10.21 . 13C NMR  δ 152.8, 151.7, 146.2, 110.2 ppm. IR (KBr) ν̃ = 3344, 3247, 3194, 1660, 1594, 1561, 1507, 1472, 1417, 1390, 1376, 1359, 1333, 1262, 1233, 1136, 989, 879, 855, 812, 778, 767, 735, 719, 713, 660, 643, 608, 580, 549, 460 cm−1. Elemental analysis (%) for C4H2N8O5 (242.01) Calcd: C 19.84, H 0.83, N 46.28. Found: C 19.43, H 1.27, N 46.64.To a 50 mL flask containing 3 ml 100% HNOSupplementary InformationPeer Review File


Eucommia ulmoides leaves (EFEL) on PM2.5-induced cognitive impairment in BALB/c mice. EFEL improved PM2.5-induced cognitive decline by improving spontaneous alternative behavioral and long-term memory ability. EFEL increased ferric reducing activity power (FRAP) in serum. In addition, EFEL increased superoxide dismutase (SOD) and reduced glutathione (GSH) contents and inhibited the production of malondialdehyde (MDA) in lung and brain tissues. EFEL also restored the mitochondrial function by regulating reactive oxygen species (ROS) production, mitochondrial membrane potential (MMP) level, and ATP level in lung and brain tissues. EFEL ameliorated the cholinergic system by regulating the acetylcholine (ACh) content and acetylcholinesterase (AChE) activity in the brain tissue and the expression of AChE and choline acetyltransferase (ChAT) in the whole brain and hippocampal tissues. EFEL reduced PM2.5-induced excessive expression of inflammatory protein related to the lung, whole brain, olfactory bulb, and hippocampus. Physiological compounds of EFEL were identified as 5-O-caffeolyquinic acid, rutin, quercetin, and quercetin glycosides. As a result, EFEL has anti-inflammation and anti-amnesic effect on PM2.5-induced cognitive impairment by regulating the inflammation and inhibiting the lung and brain tissue dysfunction, and its effect is considered to be due to the physiological compounds of EFEL.This study aimed to evaluate the protective effect of the ethyl acetate from  The PM2.5 deposited in the respiratory system causes reactive oxygen species (ROS), oxidative stress, inflammation, and apoptosis, inducing respiratory diseases such as asthma, pneumonia, chronic cough, disruption of calcium homeostasis, and mediating of inflammatory responses [2.5 exposure increases the ROS and inflammation factors by oxidative stress and inflammatory response in lung tissue [2.5 that has penetrated into the alveoli can pass through the air-blood barrier and circulate throughout the body, where it can be deposited in the brain tissue [2.5 directly reaches the olfactory bulb, passing through the blood-brain barrier (BBB), which induces nervous system dysfunction and causes neuroinflammation and neurodegenerative diseases [2.5 absorbed induces an excessive inflammatory response, which leads to cell apoptosis in the brain by producing ROS and inflammatory cytokines [Air pollution is an environmental issue that is becoming a global problem, and its harmful effect on the human body is gradually increasing . Accordidiameter . PM2.5 cg tissue . Also, Pn tissue . Also, in tissue . As a ren tissue . In addiytokines .Eucommia ulmoides is a plant in the family Eucommiaceae and is widely cultivated in China, Japan, and Korea [Eucommia ulmoides has various compounds such as flavonoids, lignans, iridoids, and phenols, and its barks and leaves are used as herbal medicines for the prevention of hypertension, diabetes, and hepotoxicity [Eucommia ulmoides leaves extract decreased the levels of total cholesterol, LDH cholesterol, and free fatty acids in serum and decreased the activity of hepatic fatty acid synthase and HMG-CoA enzyme in a high-fat diet-induced hamster model [Eucommia ulmoides leaves showed antioxidant activity in the liver of a CCl4-induced hepatotoxicity model and decreased GOT, GPT, LDH, and ALP levels in serum [Eucommia ulmoides extract improved motor dysfunction in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP-)-induced Parkinson's model mice through pole and rotarod tests and increased the levels of dopamine and its metabolites such as 3,4-dihydroxyphenylacetic acid and homovanillic acid in striatum. Also, Eucommia ulmoides extract showed a protective effect on neuroinflammation by reducing the levels of TNF-α, IL-1β, and IL-6 in the blood [Eucommia ulmoides improved the neurotoxicity of amyloid beta-induced-PC12 cells [Eucommia ulmoides leaves are mainly focused on the improvement effect of hepatoxicity and lipid metabolism, especially there are few studies on Eucommia ulmoides leaves related to the effect on PM2.5-induced inflammation and cognitive dysfunction. Therefore, in this study, anti-inflammatory and amnesic effects were confirmed using the ethyl acetate fraction from Eucommia ulmoides leaves in PM2.5-induced BALB/c mice, and the potential availability as a functional food material for the prevention of PM2.5-induced cognitive impairment was evaluated.nd Korea . Eucommitoxicity –15. Accoer model . The wathe blood . In addi12 cells . HoweverEucommia ulmoides leaves were purchased from Yeongcheon, Gyeongsangbuk-do, Korea, in April 2019 and verified by the National Institute of Forest Science . The sample was extracted in 40% ethanol for 2 h at 40°C and evaporated using a vacuum rotary evaporator . After that, the extract was sequentially fractionated using n-hexane, chloroform, and ethyl acetate and lyophilized. According to previous study, ethyl acetate fraction from Eucommia ulmoides leaves (EFEL) was presented as the significant total phenolic and flavonoid contents and antioxidant activities, and it was used for this study , EFEL 20 (PM2.5 exposure + EFEL 20 mg/kg of body weight), and EFEL 40 (PM2.5 exposure + EFEL 40 mg/kg of body weight) . PM2.5, EFEL 20, and EFEL 40 groups were exposed to PM2.5 at a constant concentration (500 μg/m3) in whole body exposure chamber, and the control group was exposed to filtered air the same time. Clean air and PM2.5 were exposed 5 hours per day for 12 weeks. EFEL was dissolved in clean drinking water according to the dose and ingested in mice by oral administration once a day for 12 weeks.The experimental animals  were bred in certain temperature (22 ± 2°C) and humidity (50 ± 5%) conditions. All experiments were performed according to the guidelines of Animal Care and Use Committee of the Gyeongsang National University . Mice were randomly divided to 4 groups: control (clean air exposure), PMThe Y-maze consists of the length (33 cm), height (15 cm), and width (10 cm), respectively. Mice were placed at the end of one arm, and the movement of each mouse was recorded by a smart video tracking system  for 8 min , 21. In 2 inhalation.Morris water maze consists of stainless circular pool (90 cm in diameter and 30 cm deep) and divided into quadrants  with visual clues, and a platform was placed in the center of the W zone. In Morris water maze test, evaluated behavioral parameters were escape times for platform in W zone of pool through hidden tests and retention time in W zone through probe test. In the hidden test, the escape time to the platform was measured for 4 days (maximum time: 60 sec). In the probe test, the platform was removed retention time in the W zone, and movements of mice were recorded using a smart video tracking system  , 22. Aftg for 10 min at 4°C was used to measure ferric reducing/antioxidant power (FRAP). Serum was reacted with FRAP reagent consisting of 10 mM 2-4-5-tripyridyl-S-triazine (TPTZ): 20 mM FeCl3: 0.3 M sodium acetate buffer (pH 3.6) (1 : 1 : 10), and then, the reaction was measured by a microplate reader at 593 nm.After the behavioral test, the serum obtained by centrifugation at 10,000×The lung and brain homogenates using 10 volumes phosphate buffer saline  were used for evaluation of antioxidant system such as superoxide dismutase (SOD) and malondialdehyde (MDA) contents, and cholinergic system, such as acetylcholine (ACh) and acetylcholinesterase (AChE), and homogenate using 10 mM phosphate buffer with 1 mM EDTA (pH 6.0) were used for evaluation of reduced glutathione (GSH) content.g for 10 min at 4°C was extracted using 10 volumes of cell extraction buffer. The extracted samples were centrifuged at 14,000×g for 5 min at 4°C, and the supernatants were used for the experiment according to manufacturer's protocol . The absorbance was measured at 450 nm  [The supernatant obtained by centrifuging the lung and brain homogenates at 400×VT, USA) , 23.g for 15 min at 4°C of lung and brain homogenate were mixed with 5% metaphosphoric acid (1 : 1), and the mixtures were centrifuged at 2,000×g for 2 min at 4°C. The supernatants were mixed with 0.26 M tris-HCl, 0.65 N NaOH, and 1 M o-phthaldialdehyde and reacted for 15 min. Fluorescence was measured by a fluorescence microplate reader  at emission (420 nm) and excitation (320 nm) wave [The supernatants obtained by centrifuging at 10,000×nm) wave , 24.The supernatants obtained by centrifuging at 5,000 rpm for 10 min at 4°C of lung and brain homogenate were reacted with 1% phosphoric acid and 0.67% TBA in water bath for 95°C for 1 h. The mixtures were spun down, and the absorbance of supernatant was measured at 532 nm  , 25.g for 30 min at 4°C was used for ACh levels and AChE activity [3 were added, and the absorbance was measured at 540 nm using microplate reader .The supernatant obtained by centrifuging the brain tissue homogenate at 14,000×activity , 26. ForTo evaluate the AChE activity, 50 mM sodium phosphate buffer was reacted with the obtained supernatant at 37°C for 15 min. After that, AChE solution composed to acetyl thiocholine and 5,5'dithio-bis (2-nitroenzoic acid) (DTNB) was added, and absorbance was measured at 405 nm using microplate reader .+), and 20 mM EDTA using a bullet blender . The pellet obtained by centrifuge  was mixed with MI buffer containing 0.1% digitonin. The mitochondrial sample and MI buffer without 20 mM EDTA were mixed and centrifuged at 13,000×g for 15 min at 4°C. The pellet obtained was mixed with the EDTA-free MI buffer and centrifuged at 10,000×g for 10 min at 4°C [Lung and brain tissues were homogenized into a mitochondrial isolation (MI) buffer including 215 mM mannitol, 75 mM sucrose, 0.1% bovine serum albumin (BSA), 20 mM HEPES (Nan at 4°C . The obtμM dichlorofluorescin diacetate (DCF-DA) reagent were mixed and reacted in the dark for 20 min. Fluorescence of mixture was measured by a fluorescence microplate reader  at emission (485 nm) and excitation (535 nm) wave.ROS content and MMP activity used quantified protein by Bradford assay . The mitμM tetraethylbenzimidazolyl-carbocyanine iodide (JC-1) in the dark for 20 min. Fluorescence of mixture was measured by a fluorescence microplate reader  at emission (535 nm) and excitation (590 nm) wave.The mitochondrial extract (1.20 mg/mL) was used to measure MMP activity, and the mitochondrial extract was reacted with EDTA-free MI buffer containing 5 mM pyruvate and malate and 1 g for 10 min at 4°C) was reacted with 1% trichloroacetic acid (TCA), and it was mixed with 25 mM Tris-acetate buffer (pH 7.7) and centrifuged at 10,000×g for 15 min at 4°C. The obtained supernatants were measured for ATP level using ATP assay kit .The mitochondrial extract was quantified at the same concentration by Bradford assay. The pellet obtained by centrifugation  according to Bradford assay. After that, supernatants were mixed with 1× loading dye and boiled at 95°C for 5 min in water bath. The proteins were separated on sodium dodecyl sulfate polyacrylamide gel (SDS-PAGE) and transferred to a polyvinylidene difluoride (PVDF) membrane . The membranes were blocked with 5% skim milk solution and incubated with primary antibody (1 : 1000) for overnight at 4°C. After incubation, the membranes were incubated with secondary antibody (1 : 2000) for 1 h at room temperature [After mice brain tissue removal, olfactory bulb and hippocampal tissue were isolated. Lung, whole brain, olfactory bulb, and hippocampus tissues were homogenized with ProtinExTM animal cell/tissue  containing 1% protease inhibitor cocktails . And homogenates were immediately centrifuged at 13,000×perature . Finally2, Vion, Waters Corp., Milford, MA, USA) was performed. EFEL was dissolved in 100% methanol and filtered using 0.2 μm membrane filters. UPLC separation was performed using an ACQUITY UPLC BEH C18 column . The flow rate was 0.60 mL/min. The mobile phases were composed as solvent A (0.1% formic acid in distilled water) and solvent B (0.1% formic acid in acetonitrile), and the analysis conditions were conducted as follows: a gradient elution of 40% A and 60% B at 0–1 min, 40% A and 60% B at 1–8 min, 100% B at 8–9 min, 100% B at 9-9.50 min, and 40% A and 60% B at 9.50–12 min. The conditions of negative electrospray ionization (ESI) were conducted as follows: ramp collision energy, 10–30 V; capillary voltage, 2.5 kV; source temperature, 100°C; desolvation temperature, 250°C; and mass range, 50–1000 m/z.To identify the physiologically compounds of EFEL, ultraperformance liquid chromatography-ion mobility separation-quadrupole time of flight/tandem mass spectrometry (UPLC-Q-TOF/MSp < 0.05) using SAS software .All experiment results were expressed as mean ± standard deviation (SD). The mean value was verified for significance differences by Duncan's multiple range test . These results indicated that all mice had the same motor performance. In the results of spontaneous alternation behavior, the PM2.5 group ) showed reduced spontaneous alternation behavior compared to the control group (40.54% ± 8.25). However, the EFEL groups showed increased spontaneous alternation behavior compared to the PM2.5 group ; EFEL 40 group: 32.33% ± 5.82 ) ) .2.5 group ) was longer than the control group (28.75 sec ± 15.87). The escape times of the EFEL groups ; EFEL 40 group, 33.78 sec ± 11.31 ) were shorter compared to the PM2.5 group. In the probe test, the retention time in the W zone of the PM2.5 group ) was decreased compared to the control group (53.80% ± 12.64). However, the EFEL groups ; EFEL 40 group, 75.16% ± 17.28 ) increased the retention time in the W zone compared to the PM2.5 group ) and improved in the EFEL groups ; and EFEL 40, 0.41 ± 0.02 ) similarly to the control group (0.40 ± 0.02).Serum FRAP activity is shown in 2.5 group ) decreased compared to the control group (4.45 ± 0.30 unit/mg of protein). However, SOD contents of the EFEL groups ; EFEL 40, 5.76 ± 0.49 unit/mg of protein ) were increased compared to the PM2.5 group ) in lung tissues of the PM2.5 group decreased compared with the control group (100 ± 24.01%). However, the EFEL groups increased the reduced GSH content ; EFEL 40, 76.62 ± 9.28% of control ) compared to the PM2.5 group ) increased compared with the control group (1.34 ± 0.04 nmole/mg of protein). However, the EFEL groups had decreased MDA production ; EFEL 40, 1.31 ± 0.08 nmole/mg of protein ) compared to the PM2.5 group ) decreased compared to the control group (4.10 ± 0.31 unit/mg of protein). It also decreased in the EFEL groups ; EFEL 40, 4.46 ± 0.36 unit/mg of protein ) compared to the PM2.5 groups ) decreased compared to the control group (100 ± 14.25%). However, the EFEL groups increased the reduced GSH content ; EFEL 40, 98.93 ± 12.55% of control ) compared to PM2.5 ) increased compared with the control group (3.50 ± 0.47 nmole/mg of protein). The EFEL groups saw decreased MDA production ; EFEL 40, 3.11 ± 0.26 nmole/mg of protein ) compared to the PM2.5 group ), and the EFEL groups ; EFEL 40, 83.35 ± 1.47% of control ) decreased the activity of AChE. In addition, PM2.5 decreased ACh content ) compared with the control group (2.18 ± 0.17 mM/mg of protein). And the EFEL groups ; EFEL 40, 1.95 ± 0.07 mM/mg of protein ) increased ACh content compared with the PM2.5 group ; hippocampus, 1.42 ± 0.01 ) compared with the control group . However, EFEL 40 decreased the AChE expression level ; hippocampus, 0.83 ± 0.20 ). Also, PM2.5 showed a reduced ChAT expression level ; hippocampus, 0.58 ± 0.21 ) compared with the control group . However, EFEL 40 increased the ChAT expression level ; hippocampus, 0.77 ± 0.17 )  and choline acetyltransferase (ChAT) is shown in  Figures . In addi Figures .2.5 group showed excessive ROS content ), while EFEL 20 and 40 had decreased the ROS content  and 61.11 ± 25.64% of control ) ), and EFEL 20 and 40 had an increased MMP level  and 160.73 ± 3.49% of control ) ) compared with the control group (0.31 ± 0.02 nmole/mg of protein), while the EFEL 20 and 40 groups showed 0.20 ± 0.05 nmole/mg of protein  and 0.25 ± 0.03 nmole/mg of protein , respectively ) . MMP wase 0.01)) . The ATPectively .2.5 group showed excessive ROS content ), while EFEL 20 and 40 had decreased ROS content  and 73.12 ± 11.64% of control , respectively) ) and EFEL 20 and 40 have an increased MMP level  and 152.17 ± 22.89% of control , respectively) ) compared with the control group (0.32 ± 0.07 nmole/mg of protein), and the EFEL 20 and 40 groups showed 0.08 ± 0.01 nmole/mg of protein  and 0.10 ± 0.05 nmole/mg of protein , respectively (The brain of the PMctively) . MMP wasctively) . The ATPectively .2.5-exposed mice lung tissue. The expression levels of TLR4, p-JNK, p-IκBα, caspase-1, TNF-α, and IL-1β , 58.75 ± 0.29% , 42.84 ± 0.29% , 336.12 ± 0.78% , 61.37 ± 0.51% , and 93.42 ± 0.57% , respectively) in the PM2.5 group were up-regulated compared to the control group. However, the expression levels of the EFEL 40 group , 45.81 ± 0.20% , 53.04 ± 0.13% , 178.49 ± 0.93% , 55.78 ± 0.19% , and 80.04 ± 0.23% , respectively) were down-regulated compared to the PM2.5 group ; olfactory bulb, 101.53 ± 0.76% ; and hippocampus, 177.29 ± 0.46% ), p-IκBα ; olfactory bulb, 16.16 ± 0.20% ; and hippocampus, 84.52 ± 0.28% ), caspase-1 ; olfactory bulb, 114.27 ± 0.85% ; and hippocampus, 65.08 ± 0.22% ), IL-1β ; olfactory bulb, 81.84 ± 0.12% ; and hippocampus, 49.81 ± 0.23% ), and TNF-α ; olfactory bulb, 104.60 ± 0.87% ; hippocampus, 92.56 ± 0.16% ) in the PM2.5 group were up-regulated compared to the control group. However, the expression levels of p-JNK ; olfactory bulb, 82.63 ± 0.47% ; and hippocampus, 129.49 ± 0.32% ), p-IκBα ; olfactory bulb, 52.20 ± 0.24% ; and hippocampus, 63.72 ± 0.31% ), caspase-1 ; olfactory bulb, 105.39 ± 0.35% ; and hippocampus, 62.70 ± 0.33% ), IL-1β ; olfactory bulb, 86.09 ± 0.38% ; and hippocampus, 49.54 ± 0.14% ), and TNF-α ; olfactory bulb, 82.42 ± 0.34% ; and hippocampus, 75.44 ± 0.47% ) in the EFEL 40 group were down-regulated compared to the PM2.5 group , 69.75 ± 0.51% , and 56.00 ± 0.40%  in the whole brain, olfactory bulb, and hippocampus compared to the control group, respectively. And EFEL decreased the expression level ; olfactory bulb, 78.83 ± 0.31% ; and hippocampus, 65.79 ± 0.24% ) compared to the PM2.5 group. The p-tau expression level increased 101.62 ± 0.71% , 31.90 ± 0.05% , and 231.82 ± 0.57%  in the whole brain, olfactory bulb, and hippocampus compared to the control group, respectively. And EFEL decreased the expression level ; olfactory bulb, 68.36 ± 0.28% ; and hippocampus, 214.58 ± 0.34% ) compared to the PM2.5 group ; compound B, 609 m/z (RT: 3.15 min); compound C, 463 m/z (RT: 3.22 min); compound D, 505 m/z (RT:3.28 min); compound E, 489 m/z (RT: 3.44 min); and compound F, 301 m/z (RT: 3.84 min). When the main fragments were compared with a previous study, these peaks were identified as 5-O-caffeoylquinic acid (compound A) [The major physiological compounds of EFEL were identified using UPLC IMS Q-TOF/MSanalysis . The MS pound A) , rutin (pound A) , luteolipound A) , and que2.5 exposure induces oxidative stress and inflammation in respiratory system [2.5 have small diameter, it is not filtered by the nasal mucosa and is deposited in the lungs and alveoli [α, IL-6, and IFN-γ) produced by inflammation in the respiratory system, which circulate through the blood vessels throughout the body, results in systemic inflammation [2.5 and inflammatory cytokines that pass through the blood brain barrier (BBB) promote neuroinflammation, which leads to cognitive and learning deficits by neuronal damage, and PM2.5 can directly penetrate the olfactory bulb tissue through the olfactory nerve [2.5 induced cognitive dysfunction, while EFEL improved the spontaneous alternative behavior and long-term memory ability of PM2.5-induced mice by regulating the inflammation and inhibited the tissues dysfunction in lung and brain tissues. Eucommia ulmoides bark showed neuroprotective effects by improving spontaneous alternative behavior and long-term memory ability in mice with amyloid-β-induced cognitive impairment [2.5-induced cognitive dysfunction. Therefore, EFEL can be used as a material that improves cognitive dysfunction in PM2.5-induced mice.PMy system . Becauseammation . Also, Pry nerve . In thispairment . Also, rpairment , 36. In 2.5 can induce oxidative stress, which leads to a breakdown of the body's antioxidant system such as SOD and GSH. According to a study by Liu and Meng [2.5 exposure decreased SOD and reduced GSH levels in the brain, lung, liver, and kidney of mice. Also, increased ROS due to an imbalance in the antioxidant system by PM2.5 exposure can also cause direct damage to cells and tissues [2.5 can easily induce lipid peroxidation of unsaturated fatty acids in the brain tissue [2.5 reduced SOD and GSH levels and increased MDA levels in mice lung and brain tissues. However, EFEL inhibited the reduction of SOD and GSH levels and the production of MDA levels. The extract of Eucommia ulmoides bark increased the content of reduced GSH and the activity of catalase in the kidneys of cadmium-induced kidney toxicity mice [Eucommia ulmoides increased the level of SOD level and GSH content in H2O2-induced PC12 cells [Eucommia ulmoides leaves inhibited the production of MDA in the serum of streptozotocin-induced diabetic rats. Rutin increased the activities of SOD, GSH, catalase, and GPx on copper sulfate-induced brain damage of rats [2.5-induced damaged antioxidant system in lung and brain tissues.PMand Meng , the PM2 tissues . The imbn tissue . Similarity mice  and incrity mice . The mac12 cells . Also, t of rats . In addi of rats . In this2.5-induced oxidative stress and neuroinflammation induce disruption of the cholinergic system, which is a characteristic of neurodegenerative diseases such as Alzheimer's and Parkinson's diseases [2.5-induced cholinergic system dysfunction and EFEL decreased the activity of AChE and increased the content of ACh and regulated the expression of AChE and ChAT in the whole brain and hippocampus tissues. The hippocampus functions related to learning and memory ability are easily affected by cholinergic dysfunction [Eucommia ulmoides decreased AChE activity in the hippocampal and cortex of scopolamine-induced learning and cognitive impairment mice [Eucommia ulmoides leaves decreased AChE activity in the hippocampus and cortex of cadmium-induced rat [2.5-induced cholinergic dysfunction. In addition, EFEL regulated the expression level of AChE and ChAT in the whole brain and hippocampus tissues. Therefore, it was confirmed that EFEL improves cognitive impairment by regulating the cholinergic system in PM2.5-induced neurotoxicity.The cholinergic system, which plays role the of neurotransmission in learning and memory ability, is highly correlated with oxidative stress and inflammation. Disruption of the cholinergic system due to increased inflammatory response leads to an abnormal neurotransmission system through the activation of the microglia and apoptosis pathway, and it is directly related to cognitive impairment . PM2.5-idiseases . In thisfunction . In the function . Therefoent mice . In addiuced rat . Quercetuced rat . Also, quced rat . In this2.5 can damage the mitochondrial structure and ATP synthesis metabolite [2.5 increased the expression of genes that induce mitochondrial fission in the nasal mucosa of rat [2.5-induced mice. Exposure to PM2.5 decreased membrane potential and increased intracellular ROS levels in human bronchial epithelial cells (BEAS-2B) [Eucommia ulmoides reduced the ROS level and improved the loss of MMP in H2O2-induced human neuroblastoma cells (SH-SY5Y) [Eucommia ulmoides improved the sodium nitroprusside-induced reduction of MMP in PC12 cells [Eucommia ulmoides increased the SOD and ATP contents and improved the loss of MMP in the mitochondria of chloral hydrate-induced traumatic brain injury mice [2.5 exposure induced mitochondrial dysfunction of lung and brain tissue, and EFEL reduced ROS production and inhibited the loss of MMP and ATP levels in mitochondria.PMtabolite . In a pra of rat . In thisBEAS-2B) . In addiSH-SY5Y) . The rut12 cells . In addiury mice . As a re2.5 entering through the respiratory tract induces oxidative stress and inflammatory responses, which can secrete various inflammatory cytokines such as interleukin (IL)-1, IL-16, MCP-1, TNF-α, and C(cysteine)-X(noncysteine)-C(cysteine) motif (CXC) chemokines such as CXC chemokine ligand 1(CXCL1) and CXC chemokine ligand 8 (CXCL8) in lung tissue [2.5 and inflammatory cytokines circulating in the whole body pass the BBB and attack brain tissue, and PM2.5 deposited in the olfactory bulb promotes neuroinflammation [p-NFκB and p-IκBα, stimulating the secretion of inflammatory cytokines such as IL-1β and TNF-α [2.5 exposure increases the activity of poly (ADP-ribose) polymerase (PARP)-1, which promotes the formation of amyloid-β, and shows excessive activity of glial cells as evidence of neuroinflammation [2.5 increased the levels of soluble and insoluble amyloid-β and phosphorylated tau protein in mouse brain tissue [β and p-tau in the brain tissue. Polysaccharides extracted from Eucommia ulmoides leaves regulated the TLR4-NFκB pathway in hepatic ischemia-reperfusion injury mice [Eucommia ulmoides bark reduced the expression level of p-JNK, p-ERK, TNF-α, and IL-1β in LPS-induced microglial BV-2 cells [α, IL-1β, IL-6, p-ERK, and p-JNK and decreased the expression level of IκB in LPS-induced lung injury mice [Eucommia ulmoides decreased the expression levels of p-ERK, p-JNK, and p-p65 in okadaic acid-induced hippocampal neurons and reduced the expression levels of amyloid-β1-42 and phosphorylation tau protein in aged triple transgenic Alzheimer's disease model mice [p-JNK, p-IκBα, caspase-1, IL-1β, and TNF-α in the lung and the expression levels of p-JNK, p-IκBα, caspase-1, IL-1β, and TNF-α in olfactory bulb caused by PM2.5 toxicity. These results suggest that the regulation of pulmonary inflammation decreases the inflammatory factor passing through the blood, thereby reducing the expression of the neuroinflammatory factor in hippocampus tissues. In addition, EFEL reduced the expression levels of amyloid-β and p-tau in the brain tissue. Therefore, EFEL can be used as a functional food material to improve PM2.5-induced inflammatory response and cognitive impairment by regulating inflammation.PMg tissue , 59. PM2ammation , 61. Theammation . The stind TNF-α . Also, Pammation . Exposurn tissue , 66. In ury mice . Eucommi-2 cells . Also, rury mice . Quercetdel mice , 71. In Eucommia ulmoides leaves (EFEL) against PM2.5-induced excessive inflammation and cognitive impairment in BALB/c mice was confirmed. EFEL increased the level of spontaneous alternation behavior and protected the long-term memory ability of PM2.5-induced mice. EFEL inhibited the antioxidant deficit and mitochondrial damage in PM2.5-induced lung and brain tissues and attenuated cerebral cholinergic dysfunction. EFEL regulated the protein expression level related to inflammation in the lung and brain tissues. Inhibition of inflammation in lung tissue by EFEL indicates reduced neuroinflammation in the whole brain, olfactory bulb, and hippocampus, which improved the learning and memory decline in mice. EFEL inhibited damage to mitochondria and antioxidant systems in lung and brain tissues, and consequently protected against the cognitive impairment induced by PM2.5. In conclusion, it is suggested that Eucommia ulmoides leaves could be used as a material for functional food to improve PM2.5-induced cognitive impairment by regulating the inflammatory response.In this study, the protective effect of the ethyl acetate fraction of


Additionally, the cross combinations P2 × P4, P3 × P5, P3 × P8, and P6 × P7 were the most promising combinations to increase yield traits and multiple physiological parameters under water deficit conditions. Furthermore, P1, P2, and P5 were recognized as promising parents to improve grain protein content and wet and dry gluten contents under drought stress. In addition, the crosses P1 × P4, P2 × P3, P2 × P5, P2 × P6, P4 × P7, P5 × P7, P5 × P8, P6 × P8, and P7 × P8 were the best combinations to improve grain protein content under water-stressed and non-stressed conditions. Certain physiological traits displayed highly positive associations with grain yield and its contributing traits under drought stress such as chlorophyll a, chlorophyll b, total chlorophyll content, photosynthetic efficiency (Fv/Fm), proline content, and relative water content, which suggest their importance for indirect selection under water deficit conditions. Otherwise, grain protein content was negatively correlated with grain yield, indicating that selection for higher grain yield could reduce grain protein content under drought stress conditions.Water scarcity is a major environmental stress that adversatively impacts wheat growth, production, and quality. Furthermore, drought is predicted to be more frequent and severe as a result of climate change, particularly in arid regions. Hence, breeding for drought-tolerant and high-yielding wheat genotypes has become more decisive to sustain its production and ensure global food security with continuing population growth. The present study aimed at evaluating different parental bread wheat genotypes  and their hybrids under normal and drought stress conditions. Gene action controlling physiological, agronomic, and quality traits through half-diallel analysis was applied. The results showed that water-deficit stress substantially decreased chlorophyll content, photosynthetic efficiency (FV/Fm), relative water content, grain yield, and yield attributes. On the other hand, proline content, antioxidant enzyme activities , grain protein content, wet gluten content, and dry gluten content were significantly increased compared to well-watered conditions. The 36 evaluated genotypes were classified based on drought tolerance indices into 5 groups varying from highly drought-tolerant (group A) to highly drought-sensitive genotypes (group E). The parental genotypes P Triticum aestivum L.) is one of the most important cereal crops a, chloro, et al. . Accordiynthesis ,84. Furtactivity ,86. On tactivity ,88. Prolactivity ,33. SOD 2 and O2 . POD con species . CAT parYield-contributing traits are the final products of physiological processes that occur at various development stages. The remarkable reductions in yield traits under water scarcity were caused by lack of absorbed water and inhibition of cell elongation and division . FurtherGrain quality is essential in terms of the nutritional benefit and economics of wheat. It varies according to genotypes, environments, and their interaction. Ozturk, et al.  elucidat3 and P7 were recognized as promising parents for improving proline content under water deficit conditions. Moreover, P2, P3, P6, and P8 are good parents for most of the studied antioxidant enzymes, chlorophyll content, and relative water content. Additionally, the parents P2, P5, and P6 could be important sources of favorable genes for reducing plant height under both normal and stressed conditions. Furthermore, improving grain yield and its components could be achieved by exploiting P3, P4, and P8, which had constantly significant and positive GCA estimates under both conditions. Subsequently, these parents could inherit beneficial alleles to their progeny and improve grain yield under water-stressed conditions. Similarly, earlier reports stated the significance of employing parents with positive and high GCA estimates for improving grain yield and contributed traits under drought stress conditions [8 exhibited favorable GCA estimates for grain yield and was also an excellent combiner for chlorophyll b, total chlorophyll content, relative water content, CAT activity, spike length, and 1000-grain weight. In consequence, it can be employed for breeding drought-tolerant and high-yielding genotypes in the wheat breeding program. The parents P1, P2, and P5 were the best combiners for improving grain protein content, and P1 was the best for improving wet and dry glutens under both conditions. Subsequently, the valuable alleles of these genotypes could be employed in breeding programs for improving grain quality under well-watered and stressed conditions. Parental genotypes with desirable GCA effects for specific traits could be valuable for providing pure lines for breeding purposes.A successful breeding program depends on the selection of suitable parental genotypes ,99,100. nditions ,103,104.2 × P4, P3 × P5, P3 × P8, and P6 × P7 were identified as excellent combiners for developing high-yielding genotypes under stressed conditions. Out of the aforementioned crosses, three hybrids, P2 × P4, P3 × P5 and P3 × P8, exhibited desirable SCA and high grain yield. The relationship between high yield performance and desired SCA effects was also deduced by Semahegn, et al. [1 × P4, P2 × P3, P2 × P5, P2 × P6, P4 × P7, P5 × P7, P5 × P8, P6 × P8, and P7 × P8 were the most merit-specific combiners for protein content. Additionally, P1 × P6, P2 × P3, P2 × P4, P3 × P7, P4 × P6, P5 × P6, and P5 × P7 appeared as the best specific combiners for wet and dry gluten under drought and non-stressed conditions. These crosses could be used to develop new genotypes with higher protein and gluten levels for specific industrial applications, as suggested by Joshi, et al. [3 × P8 was a good combiner for chlorophyll b, total chlorophyll content, Fv/Fm, RWC, proline content, CAT, POD, spike length, number of grain per spike, 1000-grain weight, and grain yield per plant. Moreover, P3 × P5 and P6 × P7 were the best combiners for chlorophyll b, total chlorophyll content, RWC, spike length, number of grain per spike, grain yield per plant, and wet gluten content. Subsequently, these crosses could be effectively exploited in wheat breeding programs to ameliorate these traits under drought stress and normal conditions [Hybrids with significant SCA effects are great choices for selection. The result revealed that the crosses P, et al.  and Kama, et al. . Grain q, et al. . Grain p, et al. ,106. The, et al. . The majnditions .3, P8, P3 × P8, P4 × P8, P3 × P5, P6 × P7, and P5 × P6 were classified to be drought-tolerant  varied from highly tolerant to highly sensitive genotypes. The genotypes Ptolerant . These gnditions ,112,113.Both additive and non-additive gene actions were included in the inheritance of all evaluated traits under both treatments, as evidenced by the highly significant GCA and SCA effects for all the evaluated traits. However, the GCA/SCA ratio was greater than unity for the majority of the evaluated characteristics in both stressed and non-stressed conditions. This implies the preponderance of additive gene effects in controlling the inheritance of these traits. These findings coincide with those of Sinolinding and Chowdhry , Farshada, chlorophyll b, Fv/Fm, relative water content, and proline content under drought stress. Accordingly, selection for improving these physiological traits under water deficit conditions will result in improving the grain yield. These findings coincide with previous reports that reflected the importance of physiological attributes as indicators for grain yield under drought stress [Understanding the associations among physiological, agronomic and quality traits could enhance breeding programs efficiency. The PC-biplot is a suitable statistical approach to assess the interrelationships among evaluated traits. The results displayed that grain yield was positively correlated with chlorophyll t stress ,118,119.t stress ,120. On t stress  Tari [12t stress , Thungo,t stress , and Šípt stress  depicted3 and P8 and their cross combination are proposed for breeding high-yielding and drought-tolerant wheat genotypes. Additionally, the parental genotypes P1, P2, and P5, as well as the hybrid combinations P1 × P4, P2 × P3, P2 × P5, P2 × P6, P4 × P7, P5 × P7, P5 × P8, P6 × P8, and P7 × P8, were the most promising genotypes for improving grain quality traits under drought-stressed conditions. The tolerance indices and cluster analysis provide valuable information on classifying the genotypes based on their tolerance to drought stress. Chlorophyll a, chlorophyll b, photosynthetic efficiency, proline content, and relative water content displayed highly positive associations with grain yield and its contributing traits under drought stress. These findings suggest the importance of these traits for indirect selection under water deficit conditions.Water-deficit stress substantially reduced all assessed traits except proline content, antioxidant enzyme activities , grain protein content, wet gluten content, and dry gluten content, which were significantly increased. The parental genotypes P


C. beijerinckii NCIMB 8052 for the fermentation of LB-derived hydrolysates to acetone–butanol–ethanol (ABE). The engineered microbial strain (C. beijerinckii_SDR) was produced by the integration of an additional copy of a short-chain dehydrogenase/reductase (SDR) gene (Cbei_3904) into the chromosome of C. beijerinckii NCIMB 8052 wildtype, where it is controlled by the constitutive thiolase promoter. The C. beijerinckii_SDR and C. beijerinckii NCIMB 8052 wildtype were used for comparative fermentation of non-detoxified and detoxified hydrothermolysis-pretreated switchgrass hydrolysates (SHs) with and without (NH4)2CO3 supplementation. In the absence of (NH4)2CO3, fermentation of non-detoxified SH with C. beijerinckii_SDR resulted in the production of 3.13- and 2.25-fold greater quantities of butanol (11.21 g/L) and total ABE (20.24 g/L), respectively, than the 3.58 g/L butanol and 8.98 g/L ABE produced by C. beijerinckii_wildtype. When the non-detoxified SH was supplemented with (NH4)2CO3, concentrations were similar for butanol (9.5 compared with 9.2 g/L) and ABE (14.2 compared with 13.5 g/L) produced by C. beijerinckii_SDR and C. beijerinckii_wildtype, respectively. Furthermore, when C. beijerinckii_SDR and C. beijerinckii_wildtype were cultured in detoxified SH medium, C. beijerinckii_SDR produced 1.11- and 1.18-fold greater quantities of butanol and ABE, respectively, than when there was culturing with C. beijerinckii_wildtype. When the combined results of the present study are considered, conclusions are that the microbial strain and medium modifications of the fermentation milieu resulted in greater production of fuels and chemicals from non-detoxified LB hydrolysates.The presence of lignocellulose-derived microbial inhibitory compounds (LDMICs) in lignocellulosic biomass (LB) hydrolysates is a barrier to efficient conversion of LB hydrolysates to fuels and chemicals by fermenting microorganisms. Results from this study provide convincing evidence regarding the effectiveness of metabolically engineered  Clostridium species can ferment a wide variety of substrates such as starch, disaccharides, hexoses, pentoses, glycerol, cellulose, and syngas into industrially important chemicals and environmentally compatible fuels  production in the exponential phase (acidogenesis) and solvent production  in the second (stationary phase). During acidogenesis, there is the production of H2, CO2, acetate, and butyrate, resulting in a decrease in culture pH. During solventogenesis, there is a marked change in metabolism resulting in the uptake of acids produced during the acidogenic phase along with sugar substrates, which are metabolized into acetone, butanol, and ethanol  such as energy crops, agricultural residues, and farm wastes—for biofuel production is being considered a “panacea” for preventing or reducing prospective competition between food sources and industrial raw materials for biofuels production . LB is cClostridium species for increased tolerance to LDMICs is viewed as a possible strategy to markedly improve efficacy and efficiency, and consequently economics of fermenting undetoxified LBH into butanol (C. beijerinckii was engineered to overexpress a short-chain dehydrogenase/reductase (SDR) H-dependent SDR when furfural was supplemented in the growth medium  in C. beijerinckii (C. beijerinckii (C. beijerinckii_SDR) however, has not been evaluated for tolerance to LDMICs in undetoxified LBH. The present study, therefore, was conducted to evaluate the capacity of C. beijerinckii_SDR to ferment undetoxified switchgrass hydrolysates (SH) to butanol.To further reduce or eliminate cost associated with detoxification of LBH prior to fermentation, metabolic engineering of solventogenic  butanol . Toward se (SDR) . This aph medium . The SDRerinckii . The resPanicum virgatum L. (Alamo switchgrass) was collected from the Gasification llaboratory at Oklahoma State University and processed using a hammer mill equipped with a 2 mm sieve. The comminuted switchgrass was pretreated in a 1 L Parr reactor  at a loading rate of 10% solids at 200°C for 10 min  using the treatment regimen described by Clostridium beijerinckii NCIMB 8052 was purchased from the American Type Culture Collection  as C. beijerinckii ATCC 51743. In an earlier study, C. beijerinckii_SDR was constructed by integrating the open reading frame Cbei_3904 (which encodes an SDR) into the chromosome of C. beijerinckii NCIMB 8052 to obtain the LDMIC-tolerant C. beijerinckii_SDR (SDR gene was constitutively expressed by placing it under the control of thiolase promoter from C. acetobutylicum ATC 824. Clostridium strains (C. beijerinckii_SDR and C. beijerinckii NCIMB 8052) were stored as spore suspensions in ddH2O at 4°C  was used to inoculate 18 ml of fresh anoxic TGY medium to increase the quantity of the preculture. These new cultures were incubated at 35°C until OD ∼ 0.9 to 1.1 was attained (3–4 h), after which they were used to inoculate the fermentation medium. All inoculations, handling, and incubation processes were performed in an anaerobic chamber  with a modified atmosphere of 82% N2, 15% CO2, and 3% H2. The TGY medium in loosely capped bottles and test tubes was maintained overnight in the anaerobic chamber to allow for the removal of residual oxygen before fermentation was initiated  was transferred into pre-sterilized 150-ml screw-cap bottles which were then supplemented with 1% (v/v) of each of the P2 buffer, mineral, and vitamin stock solutions . Sterile4)2CO3 supplementation], 44.0 ml of non-detoxified SH was transferred into another set of 150-ml Pyrex screw-cap bottles. Modified P2 buffer, mineral, and vitamin stock solutions  were added (4)2CO3 (500 µl of 200 g/L stock) was added resulting in a final concentration of 2 g/L in the fermentation medium.For fermentations using the modified medium [with  were separately inoculated into the detoxified SH fermentation medium as described previously for the non-detoxified SH fermentation medium. As with the undetoxified SH medium, another set of fermentation cultures was established with the addition of (NH4)2CO3 to a final concentration of 2 g/L. P2 medium  was used as a positive control. For the positive control, 45.5 ml of P2 medium was transferred into another set of 150-ml Pyrex screw-cap bottles. Subsequently, the P2 buffer, mineral, and vitamin stock solutions  were added using the methods described previously. The compositions of the P2 buffer, minerals, and stock solutions are presented in 600 nm, sugar, butanol, acetone, ethanol, acetic, and butyric acid concentrations.Fermentation was conducted in 150-ml Pyrex screw-cap bottles containing a 50 ml working volume of fermentation medium. ® 800 spectrophotometer  to estimate the changes in the growth of the C. beijerinckii strains during fermentation. Gas chromatography was conducted to quantify the concentrations of acetone, butanol, ethanol, acetic, and butyric acids using a 7890A system , according to a previously described method  procedure of SAS OnDemand for Academics 3.1.0 . The least square difference (LSD) test procedure was used for evaluation of treatment means. There was the determination of values for maximum optical density readings, sugar utilization during the fermentation periods, as well as maximum concentrations, yield, and production of butanol and ABE. Mean differences in these values because of using different The hydrothermolysis pretreatment method was efficacious for deconstructing switchgrass biomass. The composition of the pretreated solids has been reported by C. beijerinckii_wildtype and C. beijerinckii_SDR was rapid, attaining maximum OD600 nm of 5.7 and 5.9, respectively, in the glucose P2 medium 2CO3, there was a discernible rapid increase in the growth of C. beijerinckii_wildtype and C. beijerinckii_SDR, attaining a maximum OD600 nm of 6.8 and 7.7, respectively 2CO3 reaching OD600 nm of 5.4 and 6.2, respectively 2CO3 supplementation, the pH of the fermentation medium decreased to less than 5.4  at 12 h with a concomitant increase in acetic 2CO3, the pH for both C. beijerinckii_wildtype and C. beijerinckii_SDR increased after 12 h of fermentation (C. beijerinckii_wildtype during fermentation of non-detoxified SH negatively affected the cells (in addition to the effects of the LDMICs), the SH was detoxified by utilizing activated carbon to reduce the concentrations of LDMICs 2CO3 supplementation. Similarly, when C. beijerinckii_wildtype and C. beijerinckii_SDR were grown in detoxified SH medium supplemented with (NH4)2CO3, the pH of the cultures decreased and later increased as fermentation progressed 2CO3 supplementation, detoxified SH, and detoxified SH with (NH4)2CO3 supplementation. While the LDMIC-tolerant C. beijerinckii_SDR readily utilized the glucose in non-detoxified SH as a carbon source for energy metabolism and ABE production, such that greater than 57 g/L glucose was utilized in 84 h, the C. beijerinckii_wildtype was not efficient in utilizing glucose in non-detoxified SH  to 46 g/L 2CO3 supplementation, both microbial strains utilized similar quantities of glucose at a similar rate (C. beijerinckii_SDR was greater than that of C. beijerinckii_wildtype (C. beijerinckii_SDR utilized 42.4 g/L glucose in non-detoxified SH with (NH4)2CO3 supplementation, resulting in 1.3-fold less glucose utilized in non-detoxified SH without (NH4)2CO3 supplementation. The rate of glucose utilization by C. beijerinckii_SDR in a non-detoxified SH medium with (NH4)2CO3 supplementation was similar (0.59 g/L/h) to that of C. beijerinckii_wildtype (4)2CO3, the overall glucose utilization by C. beijerinckii_SDR and C. beijerinckii_wildtype was also similar. C. beijerinckii_SDR and C. beijerinckii_wildtype utilized approximately 48 g/L and 47 g/L glucose, respectively, at the rates of ∼0.68 and ∼0.66 g/L/h, respectively 2CO3 supplementation, detoxified SH, and detoxified SH with (NH4)2CO3 supplementation). Notably, the quantities of acetone produced by C. beijerinckii_SDR were greater than that produced by C. beijerinckii_wildtype. Acetone concentration in the medium peaked at 8.0 g/L during fermentation of non-detoxified SH without (NH4)2CO3 supplementation by C. beijerinckii_SDR, which was 2.5-fold greater than that (3.24 g/L) produced by C. beijerinckii_wildtype (4)2CO3, the concentration of acetone in the medium was similar for both fermentations with C. beijerinckii_SDR and C. beijerinckii_wildtype (4)2CO3 supplementation, there were greater acetone concentrations in the medium inoculated with C. beijerinckii_SDR than in C. beijerinckii_wildtype cultures. With C. beijerinckii_SDR, acetone concentrations were 4.4 g/L (4)2CO3 supplementation, respectively. With C. beijerinckii_wildtype, acetone concentrations in the detoxified SH with and without (NH4)2CO3 supplementation media were 3.4 g/L (4)2CO3 supplementation and in P2 medium (4)2CO3 supplementation, both strains of C. beijerinckii exhibited similar ethanol profiles until 48 h. At the 60-h time point in cultures of C. beijerinckii_SDR, there was a decrease in ethanol concentration and subsequently, an increase at 72 h, while in cultures of C. beijerinckii_wildtype there was a sustained increase in ethanol concentration up to 96 h of fermentation 2CO3 supplementation and detoxified SH with (NH4)2CO3 supplementation were similar , when the substrate was non-detoxified SH with no (NH4)2CO3 supplementation, which is 3.1- and 2.2-fold greater than the maximum butanol and ABE concentrations observed with C. beijerinckii_wildtype (C. beijerinckii_SDR in non-detoxified SH medium is greater than that it produced in P2 medium control (16.1 g/L ABE), the associated ABE productivity of 0.24 g/L/h (C. beijerinckii_SDR 84 h to produce a maximum ABE of 20.23 g/L in non-detoxified SH medium without (NH4)2CO3 supplementation, it took C. beijerinckii_SDR only 48 h to achieve maximum ABE concentration of 16.1 g/L in P2 medium control. When the non-detoxified SH medium was supplemented with (NH4)2CO3, however, C. beijerinckii_SDR produced 9.5 g/L butanol and 14.2 g/L ABE, which are 1.03- and 1.06- fold greater than the butanol (9.2 g/L) and ABE (13.35 g/L) produced by C. beijerinckii_wildtype (4)2CO3 supplementation in non-detoxified SH medium, C. beijerinckii_SDR produced 1.2- and 1.42-fold greater butanol and ABE (4)2CO3 supplementation.Fermentation with wildtype ; Table 3 and ABE ; Table 34)2CO3 supplementation. While C. beijerinckii_wildtype produced 11.05 and 15.09 g/L butanol and ABE, respectively, during fermentation of detoxified SH with no (NH4)2CO3 supplementation, C. beijerinckii_SDR produced 12.32 and 17.86 g/L butanol and ABE, respectively, which were 1.11- and 1.18-fold, respectively, greater than the concentrations produced with C. beijerinckii_wildtype (4)2CO3, C. beijerinckii_SDR produced 17.13 and 10.73 g/L ABE and butanol, respectively, which were 1.1- and 1.05-fold greater than the respective concentrations produced with C. beijerinckii_wildtype (15.57 g/L ABE and 10.25 g/L butanol). Furthermore, C. beijerinckii_SDR produced 9.99 and 16.1 g/L butanol and ABE, respectively, while C. beijerinckii_wildtype produced 9.66 and 14.08 g/L butanol and ABE, respectively, during the fermentation of the control P2 medium  to convert non-detoxified SH to butanol and ABE. Additionally, we assessed whether modification of the fermentation medium by (NH4)2CO3 supplementation results in a greater metabolism of the SH-derived glucose to butanol. The SDR superfamily of proteins consists of NAD(P)H-dependent oxidoreductases involved in a single-step reduction of aldehydes such as furans to less toxic alcohols 2CO3 supplementation. The lower cell density in cultures of C. beijerinckii_wildtype is an indication that there was marked inhibition of growth by the LDMICs present in the undetoxified SH. This contrasted with the profiles of C. beijerinckii_SDR grown in undetoxified SH  during fermentation of non-detoxified SH with (NH4)2CO3 supplementation further underscored its potential.There were 2.3-, 2.2- and 3.1-fold greater cell population, total ABE, and butanol concentrations, respectively, by the ified SH . The extn the SH . This isf LDMICs  during gf LDMICs . It is pentation . This isckii_SDR , 8A. AltC. beijerinckii_SDR and C. beijerinckii_wildtype in non-detoxified SH, with and without (NH4)2CO3 supplementation alongside the acid production profiles, the inhibitory actions of LDMICs (C. beijerinckii_SDR (9.74 g/L) or C. beijerinckii_wildtype (7.92 g/L) were greater than the acetic acid concentrations (∼3–4 g/L) typically present at the initiation time point of ABE fermentation. These relatively lesser acetic acid concentrations at 0 h are desired to maintain a pH range that supports microbial population growth and ABE production. In the present study, the acetic acid produced during pretreatment of switchgrass and that from the buffer component of the P2 medium resulted in a greater acetic acid concentration of the SH fermentation medium at 0 h 2CO3 may have contributed to the enhanced growth observed for C. beijerinckii_SDR and C. beijerinckii_wildtype during fermentation of non-detoxified SH 2CO3 had a negative effect on ABE production in cultures inoculated with C. beijerinckii_SDR. Consequently, there was 1.2- and 1.4-fold less butanol and ABE production (C. beijerinckii_SDR in non-detoxified SH with (NH4)2CO3 supplementation than in non-detoxified SH medium without (NH4)2CO3 supplementation 2CO3 supplementation 2CO3 supplementation. These conditions resulted in the maximum ABE production of 20.24 g/L (C. beijerinckii_SDR maintained pH above 5.5 in the non-detoxified SH medium supplemented with (NH4)2CO3 during solventogenic phase of the fermentation (4)2CO3 resulted in a sustained pH greater than 5.5, which led to enhanced growth of C. beijerinckii_SDR that was, in turn, sub-optimal for ABE production. Hence, there was a lesser ABE production with C. beijerinckii_SDR in non-detoxified SH medium with (NH4)2CO3 supplementation.While these larger cell populations led to 2.6- and 1.5-fold increase in butanol and ABE production, respectively, for oduction , respectentation . These fentation  where suentation . For ABEoduction . It shouoduction . During oduction . The pH 0.24 g/L . Conversentation . It is pC. beijerinckii_SDR and C. beijerinckii_wildtype produced higher butanol and ABE concentrations in detoxified SH without (NH4)2CO3 supplementation than in the P2 medium  in C. beijerinckii conferred some resiliency on the strain, in conditions that were inhibitory to C. beijerinckii_wildtype. Although the protein product of Cbei_3904 has been re-annotated as a tri/tetra-hydroxynaphthalene reductase-like enzyme, it is still an oxidoreductase and SDR superfamily domains . The tri/tetra-hydroxynaphthalene reductases are specifically involved in fatty acid biosynthesis, co-factor (biotin) metabolism, and reduction of alternate phenolic compounds and cyclic ketones in bacteria H for butanol production. Integration of the Cbei_3904 into the genome of C. beijerinckii under the control of a constitutive promoter (thiolase) ensures continued expression of the associated enzyme during both the acidogenic and solventogenic phases of growth  metabolism, and NAD(P)H-consuming reduction of phenolic compounds H utilization for different processes, with potential beneficial effect on non-NAD(P)H-dependent acetone production, which facilitates the ability of the cell to reabsorb and convert acetic acid to a neutral product (acetone).The large quantities of acetone produced by  control  are notef growth . As a NAompounds , may be  and CO2 , and incerinckii  due to tycolysis . In the ycolysis  and to sC. beijerinckii NCIMB 8052 (C. beijerinckii_SDR) to tolerate the deleterious effects of LDMICs and produce ABE with hydrothermolysis-pretreated SH was evaluated. Additionally, the effect of fermentation medium modification by (NH4)2CO3 supplementation on the fermentation profile of C. beijerinckii_SDR was investigated. Supplementation of SH with (NH4)2CO3 led to improved growth of C. beijerinckii strains and ABE production. Use of non-detoxified SH along with metabolically engineered C. beijerinckii_SDR to produce ABE is a promising combination as it produced 1.26-fold (20.24 g/L ABE) more ABE than the positive control (16.1 g/L ABE). This result underscores the robustness of C. beijerinckii_SDR for ABE production using undetoxified hydrothermolysis-pretreated SH. Moreover, our results make a case for metabolic engineering as a tool for rewiring metabolic networks in fermenting microorganisms toward enhanced production of fuels and chemicals using cheap biomass substrates.In the present study, the capacity of a genetically engineered strain of