Hand-Powered Point-of-Care: Centrifugal Microfluidic Platform for Urine Routine Examination (µCUREX).

Urinalysis is one of the simplest and most common medical tests in modern cities. With the assistance of professional technicians and equipment, people in metropolitan areas can effortlessly acquire information about their physiological conditions from traditional clinical laboratories. However, the threshold, including precise benchtop equipment and well-trained personnel, still remains a considerable dilemma for residents in healthcare-poor areas. Hence, it is a crucial and urgent topic to develop a smart and affordable widget to address this challenge. To improve the healthcare rights of residents, we proposed a disposable centrifugal microfluidic urine routine examination platform (named µCUREX) actuated with a modified hand-powered fan. Two parts of urinalysis (sediment test and chemical strip test) were integrated into the µCUREX disc. The influence on sedimentation by variant hand-powered manipulation was simulated using COMSOL. As a result, more than 70% of the sediment can be collected. Moreover, the color change of chemical strip papers (indicators for glucose, pH, protein, and occult blood) was recorded with a 3D-printed studio and analyzed after reaction with chemical-spiked and pH-adjusted artificial and human urine specimens. The whole process can be completed within 10 min, with only 200 µL of urine needed. In conclusion, we successfully constructed an ultra-low-cost point-of-care platform for urinalysis in extremely resource-poor settings. The handy size, high affordability, and user-friendliness of the µCUREX disc provide strong potential and feasibility in solving problems in resource-poor settings. Furthermore, we highly expect the µCUREX platform to improve the level of healthcare in resource-limited areas.

Effect of the Brief Instructional Video Intervention on the Quality of Cardiopulmonary Resuscitation.

Background: Chest compressions are the basis of cardiopulmonary resuscitation (CPR), and high-quality chest compressions can improve survival rate in patients with out-of-hospital cardiac arrest. Although many efforts have been made to improve the quality of CPR in inexperienced adults, the results are still not high, especially during emergencies. The primary purpose of this study is to investigate whether a brief instructional chest compression-only CPR video could improve chest compression quality in inexperienced adults. Methods: One hundred adults with no CPR experience (age: 20.28 ± 2.28 years; women: 50, men: 50) participated in this study. Participants completed body composition and handgrip strength measurements, and performed two CPR quality tests on the Laerdal® Little Anne QCPR Manikin, namely without video-CPR (WV-CPR) and video-CPR (V-CPR). The WV-CPR quality test was performed first. After 2 minutes of continuous chest compression, the participants rested for 10 seconds and repeated 3 cycles (phase 1, phase 2, and phase 3). After resting for more than 72 hours, V-CPR quality test was conducted. During the V-CPR with video intervention, the participants also continued to compress the chest for 2 minutes, and then rested for 10 seconds, repeating 3 cycles. Results: In phase 1, compared with WV-CPR, the V-CPR has a significant increase (p < 0.001) in chest compression fraction (CCF) (56.31 ± 33.22% vs. 41.82 ± 32.30%) and percent of correct compression rate (PCCR) (96.17 ± 8.45% vs. 26.31 ± 37.55%). In addition, the V-CPR has significantly lower (p < 0.001) chest compression rate (CCR) (110.85 ± 2.40 cpm vs. 128.86 ± 24.52 cpm) and rating of perceived exertion (RPE) (11.89 ± 2.25 vs. 12.87 ± 2.25). For phases 2 through 3, V-CPR and WV-CPR achieved significant differences in CCF, CCD, CCR, PCCR, and RPE (p < 0.01). There were significant differences (p < 0.05) in CCF, CCD, chest compression rebound rate, and RPE among the different administration stages of both WV-CPR and V-CPR. Conclusions: The results of this study revealed that a brief instructional chest compression-only CPR video could improve chest compression quality for inexperienced adults by reducing fatigue and CCR, and increasing CCF and PCCR.

Synthesis of cinnamils and quinoxalines and their biological evaluation as anticancer agents.

We synthesized multiple cinnamils and quinoxalines to evaluate their anticancer activity. Cinnamils were used as precursors for quinoxalines via condensation with 1,2-diaminobenzene. Among the 26 synthesized compounds reported in this article, we found that cinnamil 3l exhibited its inhibitory effect with an IC50 value of 1.45 ± 0.98 µM, significantly higher than doxorubicin (8.5 ± 0.85 µM) against pancreatic cancer cells (PANC-1). Additionally, cinnamil 3l (IC50 10.98 ± 3.63 µM) showed less cytotoxicity than doxorubicin to Hs68 cells (0.92 ± 1.11 µM). The colony formation assay demonstrated that 3l obviously decreased the PANC-1 cell viability, and Western blot assays confirmed that 3l markedly induced apoptosis of PANC-1 cells through Bax, Bcl-2, and caspase 3 signaling cascades. These results demonstrate that cinnamil 3l has great potential to be further developed as a promising chemotherapeutic agent for pancreatic cancer.

Evaluation of the Efficacy of Supplementation with Planox® Lemon Verbena Extract in Improving Oxidative Stress and Muscle Damage: A Randomized Double-Blind Controlled Trial.

Excessive exercise load can cause muscle soreness and fatigue, as well as inflammation and oxidative stress. Lemon verbena (Aloysia triphylla; Lippia citriodora) is often used as a spice in tea or beverages. Its leaves are rich in polyphenols, which have antioxidant and anti-inflammatory bioactivities. In the present study, we investigated whether supplementation with Planox® lemon verbena extract (LVE) could improve muscle damage and biochemical indicators after exhaustive exercise challenge. All subjects (30 males and 30 females) underwent a double-blind trial and were randomly divided into a placebo group (0 mg/human/day) and an LVE supplement group (400 mg/human/day), with gender-equal distribution. All subjects started supplementation 10 days before exhaustive exercise and continued it until all tests were completed. Before the intervention, after the exhaustive exercise, and on the following 3 days, the participants underwent 12-minute Cooper running/walking; blood collection; assessments of pain, muscle stiffness, maximum jump heights, and isometric maximum muscle strength. The results showed that supplementation with LVE effectively increased GPx and reduced CK, IL-6, 8-OHdG and muscle pain after the exhaustive exercise, but it had significant effect on strength recovery. In summary, LVE is a safe and edible natural plant extract that can reduce muscle damage and soreness after exercise. This trial was registered at clinicaltrials.gov as NCT04742244.

The feasibility of emergency medical technicians performing intermittent high-quality cardiopulmonary resuscitation.

Background: Whether intermittent chest compressions have an effect on the quality of CPR is worthy of discussion. The purpose of this study was to investigate differences in the chest compression quality of emergency medical technicians (EMTs) performing cardiopulmonary resuscitation (CPR) with different rest intervals. Methods: Seventy male firefighters with EMT licenses participated in this study. Participants completed body composition measurements and three CPR quality tests, as follows: (1) CPR-uninterrupted for 10 minutes; (2) after 2 days of rest, CPR 10s-intermittent (CPR-10s), for 2 minutes each time and 5 cycles; (3) after another 2 days of rest, CPR 20s-intermittent (CPR-20s), for 2 minutes each time and 5 cycles. Results: Body composition results showed that body mass (BM), body mass index (BMI), upper limb muscle mass (ULMM), core muscle mass (CMM), and upper limb-core muscle mass (UL+CMM) were positively correlated with chest compression depth (CCD) (p < 0.05). Analysis of the three different modes of CPR quality analysis indicated significant differences in the chest compression fraction (CCF, F = 6.801, p = 0.001), chest compression rebound rate (CCRR, F = 3.919, p = 0.021), and ratings of perceived exertion (RPE, F = 23.815, p < 0.001). Among the different performance cycles of CPR-10s, significant differences were found in CCF, CCD, CCR (chest compression rate), and RPE (p < 0.05). On the other hand, among the different performance cycles of CPR-20s, significant differences were found in CCD, CCR, and RPE (p < 0.05). Moreover, the CCF, CCD, and RPE scores of the two tests reached significant differences in specific phases (p < 0.05). Conclusions: This study confirmed that the upper limb muscle mass or the weight of the upper body of EMTs is positively correlated with the quality of CPR. In addition, intermittent chest compressions with safe interruption intervals can reduce fatigue caused by long-term chest compressions and maintain better chest compression quality.

Effect of Ambulance Stretcher Bed Height Adjustment on CPR Quality and Rescuer Fatigue in a Laboratory Environment.

Background: The quality of cardiopulmonary resuscitation (CPR) is closely related to the survival rate of a patient, and it is crucial to maintain the quality of CPR during the ambulance journey to the receiving hospital. The purpose of this study was to investigate the effects of different stretcher bed heights on operator CPR quality. Methods: In this randomized crossover trial, 16 male emergency medical technicians-paramedics (EMT-Ps) performed continuous chest compressions on a hemimorphic mannequin for 5 minutes, alternating between the current height of the stretcher bed on the ambulance (38 ± 1 cm) (S-38) and the height of the participant's midpoint of the patella (S-knee), where the stretcher bed surface is. Results: According to the analysis of the quality of CPR exercises with two different stretcher bed heights at 5 minutes of continuous chest compression, the mean chest compression depth (CCD) of the S-38 position (53.81 ± 1.91 cm) was significantly lower than that of the S-knee (55.12 ± 2.03 cm; p < 0.001). The mean chest compression rate (CCR) of the S-38 position (111.44 ± 3.44 beats/min) was significantly higher than that of the S-knee (109.63 ± 4.46 beats/min; p = 0.027). The mean of total chest compressions (TCC) of the S-38 position (557.44 ± 16.81 times) was significantly higher than that of the S-knee (548.24 ± 19.40 times; p = 0.029). The rating of perceived exertion (RPE) of the S-38 position was significantly higher than that of the S-knee (12.75 ± 1.91 %; p = 0.015). Only the chest compression rebound rate (CCRR) (S-38: 97.56 ± 4.63 % vs. S-knee: 98.31 ± 1.89 %, p = 0.401) and the chest compression fraction (CCF) (S-38: 98.44 ± 0.81 % vs. S-knee: 98.44 ± 0.96 %, p = 1.000) did not reach a significant difference. Conclusion: When a resuscitator is performing chest compressions in a standing position in an ambulance, the excessive downward leaning of the resuscitator's upper body affects CPR quality and increases fatigue. This study has verified that setting the stretcher bed of the ambulance at the knee height of the EMTs provides better CPR quality and lower fatigue.

Anti-Cancer and Anti-Inflammatory Activities of Three New Chromone Derivatives from the Marine-Derived Penicillium citrinum.

Three new and uncommon chromone analogs, epiremisporine F (1), epiremisporine G (2), and epiremisporine H (3), were isolated from marine-origin Penicillium citrinum. Among the isolated compounds, compounds 2-3 remarkably suppressed fMLP-induced superoxide anion generation by human neutrophils, with IC50 values of 31.68 ± 2.53, and 33.52 ± 0.42 µM, respectively. Compound 3 exhibited cytotoxic activities against human colon carcinoma (HT-29) and non-small lung cancer cell (A549) with IC50 values of 21.17 ± 4.89 and 31.43 ± 3.01 µM, respectively, and Western blot assay confirmed that compound 3 obviously induced apoptosis of HT-29 cells, via Bcl-2, Bax, and caspase 3 signaling cascades.

Rare Chromone Derivatives from the Marine-Derived Penicillium citrinum with Anti-Cancer and Anti-Inflammatory Activities.

Three new and rare chromone derivatives, epiremisporine C (1), epiremisporine D (2), and epiremisporine E (3), were isolated from marine-derived Penicillium citrinum, together with four known compounds, epiremisporine B (4), penicitrinone A (5), 8-hydroxy-1-methoxycarbonyl-6-methylxanthone (6), and isoconiochaetone C (7). Among the isolated compounds, compounds 2-5 significantly decreased fMLP-induced superoxide anion generation by human neutrophils, with IC50 values of 6.39 ± 0.40, 8.28 ± 0.29, 3.62 ± 0.61, and 2.67 ± 0.10 µM, respectively. Compounds 3 and 4 exhibited cytotoxic activities with IC50 values of 43.82 ± 6.33 and 32.29 ± 4.83 µM, respectively, against non-small lung cancer cell (A549), and Western blot assay confirmed that compounds 3 and 4 markedly induced apoptosis of A549 cells, through Bcl-2, Bax, and caspase 3 signaling cascades.

Dibenzofuran, 4-Chromanone, Acetophenone, and Dithiecine Derivatives: Cytotoxic Constituents from Eupatorium fortunei.

Five new compounds, eupatodibenzofuran A (1), eupatodibenzofuran B (2), 6-acetyl-8-methoxy-2,2-dimethylchroman-4-one (3), eupatofortunone (4), and eupatodithiecine (5), have been isolated from the aerial part of Eupatorium fortunei, together with 11 known compounds (6‒16). Compounds 1 and 2 featured a new carbon skeleton with an unprecedented 1-(9-(4-methylphenyl)-6-methyldibe nzo[b,d]furan-2-yl)ethenone. Among the isolates, compound 1 exhibited potent inhibitory activity with IC50 values of 5.95 ± 0.89 and 5.55 ± 0.23 µM, respectively, against A549 and MCF-7 cells. The colony-formation assay demonstrated that compound 1 (5 µM) obviously decreased A549 and MCF-7 cell proliferation, and Western blot test confirmed that compound 1 markedly induced apoptosis of A549 and MCF-7 cells through mitochondrial- and caspase-3-dependent pathways.

The Relationship between the Glenohumeral Joint Internal Rotation Deficit and the Trunk Compensation Movement in Baseball Pitchers.

BACKGROUND AND OBJECTIVES: Glenohumeral joint internal rotation deficit (GIRD) is commonly observed in the dominant arm of baseball pitchers and is limited by horizontal adduction motions. We inferred that when pitchers' generation of internal shoulder rotation and horizontal adduction activity is limited, they may generate compensation movements in other body parts. This study aims to investigate whether pitchers with GIRD generates trunk compensation during pitching where pitching targets were on the lower corner of their non-dominant side. DESIGN: Case-control study. SETTING: Elite senior high school baseball. PARTICIPANTS: Twenty-five senior high school baseball pitchers participated in this study. Twelve pitchers with GIRD were assigned to the experiment group, and the remaining 13 participants to the control group. MAIN OUTCOME MEASURES: Glenohumeral internal/external rotation of both arms and internal/external rotation of the bilateral hip joints were measured. The kinematic values of the trunk when pitching to a target were measured using high-speed infrared cameras. RESULTS: Pitchers with GIRD exhibited significantly greater upper trunk rotation toward the non-dominant side when a baseball was released from their hand (27.39 ± 6.62 degrees), compared with non-GIRD pitchers (20.42 ± 5.97 degrees) (p < 0.05). The total rotation of the pivot leg of pitchers with GIRD (67.54 ± 7.84 degrees) was significantly smaller than that of pitchers without GIRD (74.00 ± 7.07 degrees) (p < 0.05). CONCLUSIONS: GIRD in the dominant arm affected upper trunk rotation during pitching and was associated with the hip range of motion. Future studies could conduct a longitudinal study regarding the relationship between GIRD and other joint injuries of the lower limbs.

Congenital exercise ability ameliorates muscle atrophy but not spinal cord recovery in spinal cord injury mouse model.

Spinal cord injury (SCI) can cause loss of mobility in the limbs, and no drugs, surgical procedures, or rehabilitation strategies provide a complete cure. Exercise capacity is thought to be associated with the causes of many diseases. However, no studies to date have assessed whether congenital exercise ability is related to the recovery of spinal cord injury. High congenital exercise ability (HE) and low congenital exercise ability (LE) mice were artificially bred from the same founder ICR mice. The HE and LE groups still exhibited differences in exercise ability after 13 generations of breeding. Histological staining and immunohistochemistry staining indicated no significant differences between the HE and LE groups on recovery of the spinal cord. In contrast, after SCI, the HE group exhibited better mobility in gait analysis and longer endurance times in the exhaustive swimming test than the LE group. In addition, after SCI, the HE group also exhibited less atrophy than the LE group, and no inflammatory cells appeared. In conclusion, we found that high congenital exercise ability may reduce the rate of muscle atrophy. This result can be applied to sports science and rehabilitation science as a reference for preventive medicine research.

PlantTalk: A Smartphone-Based Intelligent Hydroponic Plant Box.

This paper proposes an IoT-based intelligent hydroponic plant factory solution called PlantTalk. The novelty of our approach is that the PlantTalk intelligence can be built through an arbitrary smartphone. We show that PlantTalk can flexibly configure the connections of various plant sensors and actuators through a smartphone. One can also conveniently write Python programs for plant-care intelligence through the smart phone. The developed plant-care intelligence includes automatic LED lighting, water spray, water pump and so on. As an example, we show that the PlantTalk intelligence effectively lowers the CO2 concentration, and the reduction speed is 53% faster than a traditional plant system. PlantTalk has been extended for a plant factory called AgriTalk.

Accuracy of the energy expenditure during uphill exercise measured by the Waist-worn ActiGraph.

BACKGROUND/OBJECTIVE: The application of Micro-Electro-Mechanical Sensors (MEMS) as measurements of energy expenditure (EE) has certain disadvantages. For example, the inertial sensors cannot easily distinguish changes in ground slope during walking/running conditions, so the accuracy of EE calculation is biased. To resolve this issue, heart rate (HR) and heart rate reserve (HRR) were used as compensatory factors respectively to correct the classical empirical formula of the accelerometer analyzer for EE in this study. METHODS: To explore the improvement of the accuracy of EE during uphill exercise and compare the correction levels between HR and HRR, oxygen uptake was used as a criterion measure (CM). Thirty healthy adult males wore an ActiGraph GT3X with the Polar HR monitor and Vmax indirect calorimeter during twelve treadmill activities (3 gradients and 4 speeds). RESULTS: When the slopes were increased by 0%, 3%, and 6%, the measurement accuracy of the accelerometers, calculated by intraclass correlation coefficient (ICC), decreased by 0.877, 0.755, and 0.504, respectively (p < 0.05). The HR and HRR parameters of linear regression were used to correct the classical formula. The results showed that HR had higher coefficients of determination (R2) (0.801, 0.700, and 0.642 respectively) and ICCs (0.887, 0.825, and 0.785 respectively) than did the accelerometer outputs. HRR showed the highest coefficients of determination (R2) (0.821, 0.728, and 0.656 respectively) and ICCs (0.901, 0.844, and 0.795 respectively). CONCLUSIONS: Through adding HRR parameters, the accuracy of the classical prediction formula EE was significantly improved during walking/running on sloping ground.

MAGIC: an automated N-linked glycoprotein identification tool using a Y1-ion pattern matching algorithm and in silico MS² approach.

Glycosylation is a highly complex modification influencing the functions and activities of proteins. Interpretation of intact glycopeptide spectra is crucial but challenging. In this paper, we present a mass spectrometry-based automated glycopeptide identification platform (MAGIC) to identify peptide sequences and glycan compositions directly from intact N-linked glycopeptide collision-induced-dissociation spectra. The identification of the Y1 (peptideY0 + GlcNAc) ion is critical for the correct analysis of unknown glycoproteins, especially without prior knowledge of the proteins and glycans present in the sample. To ensure accurate Y1-ion assignment, we propose a novel algorithm called Trident that detects a triplet pattern corresponding to [Y0, Y1, Y2] or [Y0-NH3, Y0, Y1] from the fragmentation of the common trimannosyl core of N-linked glycopeptides. To facilitate the subsequent peptide sequence identification by common database search engines, MAGIC generates in silico spectra by overwriting the original precursor with the naked peptide m/z and removing all of the glycan-related ions. Finally, MAGIC computes the glycan compositions and ranks them. For the model glycoprotein horseradish peroxidase (HRP) and a 5-glycoprotein mixture, a 2- to 31-fold increase in the relative intensities of the peptide fragments was achieved, which led to the identification of 7 tryptic glycopeptides from HRP and 16 glycopeptides from the mixture via Mascot. In the HeLa cell proteome data set, MAGIC processed over a thousand MS(2) spectra in 3 min on a PC and reported 36 glycopeptides from 26 glycoproteins. Finally, a remarkable false discovery rate of 0 was achieved on the N-glycosylation-free Escherichia coli data set. MAGIC is available at http://ms.iis.sinica.edu.tw/COmics/Software_MAGIC.html .

Hematological and plasma profiles and ticks and tick-borne pathogens in wild Formosan black bears (Ursus thibetanus formosanus).

BACKGROUND: The endangered Formosan black bear (Ursus thibetanus formosanus) is the largest native carnivorous mammal in Taiwan. Diseases, poor management, illegal hunting, and habitat destruction are serious threats to the survival of bear populations. However, studies on the impact of diseases on bear populations are limited. Therefore, this study aimed to establish a database of the hematological and plasma profiles of free-ranging Formosan black bears and investigate the occurrence of ectoparasites, blood parasites, and vector-borne pathogens. METHODS: Formosan black bears were captured in Yushan National Park (YNP) and Daxueshan Forest Recreation Area (DSY) in Taiwan. Blood samples were collected from each bear for hematological analysis and plasma biochemistry using a hematology analyzer. Parasites and pathogens were detected using a thin blood smear with Wright-Giemsa staining and polymerase chain reaction (PCR) assay. Additionally, macroscopic ectoparasites were collected from bears to detect blood parasites and other pathogens. Moreover, the relationships between the bear variables (sex, age, and occurrence of parasites or pathogens), ectoparasites, and infectious agents were also analyzed. RESULTS: In all, 21 wild bears (14 in YNP and 7 in DSY) were captured and released during the satellite tracking studies. Hematological analysis and plasma biochemistry indicated significant differences in white blood cells (WBC), segments, creatine kinase (CK), and lactate dehydrogenase (LDH) levels between foot snare and culvert-captured bears. Additionally, there were significant differences in total plasma protein (TPP), creatinine, Ca2+, Mg2+, and K+ levels between male and female bears. Moreover, pathogen-infected bears had significantly higher erythrocyte sedimentation rate (ESR; 30 min and 1 h) and globulin levels than uninfected bears. In total, 240 ticks were collected from 13 bears, among which eight adult tick species were identified, including Haemaphysalis flava, Haemaphysalis hystricis, Amblyomma testudinarium, Ixodes ovatus, Dermacentor taiwanensis, Haemaphysalis longicornis, Ixodes acutitarsus, Amblyomma javanense, and nymphs belonging to Haemaphysalis spp. PCR revealed that 13 (61.90%) and 8 (38.10%) bears harbored Hepatozoon ursi and Babesia DNA, respectively. Among the ticks examined, 157 (65.41%) and 128 (53.33%) samples were positive for H. ursi and Babesia, respectively. CONCLUSIONS: To the best of our knowledge, this is the first study to establish a database of the hematological and plasma profiles of wild Formosan black bears and investigate ectoparasite infestation and Hepatozoon and Babesia spp. INFECTION: In conclusion, these findings may serve as a reference for monitoring the health and population of locally endangered bears.

Acute effects of muscle mechanical properties after 2000-m rowing in young male rowers.

Background: The mechanical properties of muscles, such as changes in muscle tone and stiffness, are related to sports performance and injuries. Rowers are at increased risk of muscle fatigue and injury during high-repetition and heavy-load cyclic muscle actions. In view of this, the aim of the present study was to investigate the acute effect on muscle tone and stiffness, as well as bilateral muscle asymmetry, in high school rowers after a 2000-meter rowing ergometer test. Methods: Twelve young male rowers (age = 17.1 ± 0.9 years, body weight = 73.5 ± 9.7 kg) were included in the study. The data of muscle tone (frequency) and stiffness of the posterior deltoids (PD), latissimus dorsi (LD), and rectus femoris (RF) (dominant and non-dominant side) before and after a 2000-m rowing ergometer test were collected using a handheld MyotonPRO device. Results: After the rowing ergometer test, the muscle tone of dominant side PD, LD, and RF were significantly increased (p < 0.05). On the other hand, the muscle stiffness of the non-dominant side LD and RF, as well as the dominant side PD, LD, and RF were significantly increased after the rowing ergometer test (p < 0.05). The muscle tone and stiffness results showed that the dominant side PD, LD, and RF were all significantly higher than the non-dominant side after the rowing ergometer test (p < 0.05), where bilateral PD and RF exhibits moderate asymmetry (5% < symmetry index < 10%). Conclusions: After a high-intensity and high-load 2000-m rowing ergometer test, PD, LD, and RF showed increases in muscle tone and stiffness, as well as changes in the symmetry of bilateral muscle mechanical properties.

Rapid detection of live bacteria in water using nylon filter membrane-integrated centrifugal microfluidics.

Water is one of the most indispensable elements for human beings. People can live without food for a couple of weeks but cannot live without water for a couple of days. Unfortunately, drinking water is not always safe around the world; in many areas, the water for drinking could be contaminated with various microbes. However, the total viable microbe count in water still relies on culture-based methods in laboratories. Therefore, in this work, we report a novel, simple, and highly efficient strategy to detect live bacteria in water via a nylon membrane-integrated centrifugal microfluidic device. A handheld fan and a rechargeable hand warmer were utilized as the centrifugal rotor and the heat resource for reactions, respectively. The bacteria in water can be rapidly concentrated >500-fold by our centrifugation system. After incubation with water-soluble tetrazolium-8 (WST-8), the color change of the nylon membranes can be visually interpreted directly by the naked eye or recorded with a smartphone camera. The whole process can be finished in 3 h, and the detection limit can reach 102 CFU/mL. The detection range ranges from 102 CFU/mL to 105 CFU/mL. The cell counting results of our platform are highly positively correlated with the results of cell counting by the conventional lysogeny broth (LB) agar plate approach or the commercial 3 M Petrifilm™ cell counting plate. Our platform provides a convenient and sensitive strategy for rapid monitoring. We highly anticipate that this platform can improve water quality monitoring in resource-poor countries in the near future.

Transporter modulation of molnupiravir and its metabolite ß-D-N4-hydroxycytidine across the blood-brain barrier in a rat.

BACKGROUND: The antiviral drug molnupiravir is an orally bioavailable prodrug of the nucleoside analog ß-D-N4-hydroxycytidine (NHC), which is used to treat coronavirus disease 2019 (COVID-19). However, there is very little information on the barrier distribution of molnupiravir. Our hypothesis is that molnupiravir and NHC can penetrate the blood‒brain barrier (BBB) into brain tissue and that nucleoside transporters (equilibrative nucleoside transporters; ENT and concentrative nucleoside transporters; CNT) can modulate this process. METHODS: To investigate the mechanism of molnupiravir transport through the BBB, multiple microdialyses coupled to a validated ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC‒MS/MS) was developed to monitor dialysates, and nitrobenzylthioinosine (NBMPR; an inhibitor of ENT) was administered concomitantly with molnupiravir (100 mg/kg, i.v.) in the male rat. RESULTS: Here, we show that molnupiravir is rapidly metabolized to NHC in the blood and crossed the BBB in 20 min. Furthermore, when NBMPR is concomitantly administered to inhibit efflux, the concentrations of molnupiravir and NHC in the brain increased significantly. CONCLUSIONS: In summary, molnupiravir rapidly transforms into NHC and crosses the BBB and reaches the brain at approximately 0.3-0.8% of the blood‒brain ratio. The maximum concentration of NHC in the blood and brain is above the average half maximal inhibitory concentration (IC50) of the drug required to treat severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, suggesting a therapeutic effect. The penetration of NHC is modulated by NBMPR. These findings provide constructive information on brain disorders in clinical patients with COVID-19.

Due to the global pandemic of coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome (SARS-CoV-2), molnupiravir is used orally to treat COVID-19 with emergency use authorization. However, it is not well understood whether molnupiravir and its active component can cross the blood­brain barrier. The aim of the study was to develop an experimental mouse model to monitor the journey of molnupiravir and its active component through the bloodstream and eventually into the brain. Our experimental data suggest that a therapeutically useful amount of molnupiravir crosses from the bloodstream into the brain.

Transfer and biotransformation of the COVID-19 prodrug molnupiravir and its metabolite ß-D-N4-hydroxycytidine across the blood-placenta barrier.

BACKGROUND: Molnupiravir is an orally bioavailable prodrug of the nucleoside analogue ß-D-N4-hydroxycytidine (NHC) and is used to treat coronavirus disease 2019 (COVID-19). However, the pharmacokinetics and transplacental transfer of molnupiravir in pregnant women are still not well understood. In the present study, we investigated the hypothesis that molnupiravir and NHC cross the blood-placenta barrier into the fetus. METHODS: A multisite microdialysis coupled with a validated ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC‒MS/MS) system was developed to monitor the dialysate levels of molnupiravir and NHC in maternal rat blood and conceptus (the collective term for the fetus, placenta, and amniotic fluid). Molnupiravir was administered intravenously (100 mg/kg, i.v.) on gestational day 16. To investigate the mechanism of transport of molnupiravir across the blood-placenta barrier, we coadministered nitrobenzylthioinosine (NBMPR, 10 mg/kg, i.v.) to inhibit equilibrative nucleoside transporter (ENT). FINDINGS: We report that molnupiravir is rapidly metabolized to NHC and then rapidly transformed in the fetus, placenta, amniotic fluid, and maternal blood. Our pharmacokinetics analysis revealed that the area under the concentration curve (AUC) for the mother-to-fetus ratio (AUCfetus/AUCblood) of NHC was 0.29 ± 0.11. Further, we demonstrated that the transport of NHC in the placenta may not be subject to modulation by the ENT. INTERPRETATION: Our results show that NHC is the predominant bioactive metabolite of molnupiravir and rapidly crosses the blood-placenta barrier in pregnant rats. The NHC concentration in maternal blood and conceptus was above the average median inhibitory concentration (IC50) for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), suggesting a therapeutic effect. These findings support the use of molnupiravir in pregnant patients infected with COVID. FUNDING: This study was supported in part by research grants from the National Science and Technology Council of Taiwan (NSTC 111-2113-M-A49-018 and NSTC 112-2321-B-A49-005).

Biotransformation and brain distribution of the anti-COVID-19 drug molnupiravir and herb-drug pharmacokinetic interactions between the herbal extract Scutellaria formula-NRICM101.

The aim of this study was to explore the effects of herbal drug pharmacokinetic interactions on the biotransformation of molnupiravir and its metabolite ß-D-N4-hydroxycytidine (NHC) in the blood and brain. To investigate the biotransformation mechanism, a carboxylesterase inhibitor, bis(4-nitrophenyl)phosphate (BNPP), was administered. Not only molnupiravir but also the herbal medicine Scutellaria formula-NRICM101 is potentially affected by coadministration with molnupiravir. However, the herb-drug interaction between molnupiravir and the Scutellaria formula-NRICM101 has not yet been investigated. We hypothesized that the complex bioactive herbal ingredients in the extract of the Scutellaria formula-NRICM101, the biotransformation and penetration of the bloodbrain barrier of molnupiravir are altered by inhibition of carboxylesterase. To monitor the analytes, ultrahigh-performance liquid chromatography tandem mass spectrometry (UHPLCMS/MS) coupled with the microdialysis method was developed. Based on the dose transfer from humans to rats, a dose of molnupiravir (100 mg/kg, i.v.), molnupiravir (100 mg/kg, i.v.) + BNPP (50 mg/kg, i.v.), and molnupiravir (100 mg/kg, i.v.) + the Scutellaria formula-NRICM101 extract (1.27 g/kg, per day, for 5 consecutive days) were administered. The results showed that molnupiravir was rapidly metabolized to NHC and penetrated into the brain striatum. However, when concomitant with BNPP, NHC was suppressed, and molnupiravir was enhanced. The blood-to-brain penetration ratios were 2% and 6%, respectively. In summary, the extract of the Scutellaria formula-NRICM101 provides a pharmacological effect similar to that of the carboxylesterase inhibitor to suppress NHC in the blood, and the brain penetration ratio was increased, but the concentration is also higher than the effective concentration in the blood and brain.