Enrichment of Lactoferrin and Immunoglobulin G from Acid Whey by Cross-Flow Filtration.

The production of cream cheese, curd, high-protein yogurt, or caseinate results in large amounts of acid whey as a by-product. So far acid whey is often disposed as animal feed or organic fertilizer. However, these approaches ignore the valorization potential that arises from the unique composition of the whey protein fraction. Whey contains the biofunctional proteins lactoferrin and immunoglobulin G, which possess immune-supporting, antibacterial, antiviral, and numerous further health-promoting functions. However, the concentration of these proteins in bovine milk or whey is below a physiologically relevant level. Based on literature research we specified a daily intake of 200 mg lactoferrin as the minimal functional dose. By means of cross-flow ultrafiltration, an attempt was made to increase the concentration of biofunctional proteins. Therefore, a membrane for the selective retention of lactoferrin and immunoglobulin G was identified, and the process parameters were optimized. Finally, a concentration experiment was conducted, whereby the concentration of biofunctional proteins was increased up to factor 30. The biofunctionality was assessed in a microbiological assay. Surprisingly, the antimicrobial growth inhibition of the produced concentrate was even higher than in pure lactoferrin. The presented approach offers a strategy to convert an abundant but underutilized by-product into valuable products for human nutrition.

Dry-spun carbon nanotube ultrafiltration membranes tailored by anti-viral metal oxide coatings for human coronavirus 229E capture in water.

Although waterborne virus removal may be achieved using separation membrane technologies, such technologies remain largely inefficient at generating virus-free effluents due to the lack of anti-viral reactivity of conventional membrane materials required to deactivating viruses. Here, a stepwise approach towards simultaneous filtration and disinfection of Human Coronavirus 229E (HCoV-229E) in water effluents, is proposed by engineering dry-spun ultrafiltration carbon nanotube (CNT) membranes, coated with anti-viral SnO2 thin films via atomic layer deposition. The thickness and pore size of the engineered CNT membranes were fine-tuned by varying spinnable CNT sheets and their relative orientations on carbon nanofibre (CNF) porous supports to reach thicknesses less than 1 µm and pore size around 28 nm. The nanoscale SnO2 coatings were found to further reduce the pore size down to ∼21 nm and provide more functional groups on the membrane surface to capture the viruses via size exclusion and electrostatic attractions. The synthesized CNT and SnO2 coated CNT membranes were shown to attain a viral removal efficiency above 6.7 log10 against HCoV-229E virus with fast water permeance up to ∼4 × 103 and 3.5 × 103 L.m-2.h-1.bar-1, respectively. Such high performance was achieved by increasing the dry-spun CNT sheets up to 60 layers, orienting successive 30 CNT layers at 45°, and coating 40 nm SnO2 on the synthesized membranes. The current study provides an efficient scalable fabrication scheme to engineer flexible ultrafiltration CNT-based membranes for cost-effective filtration and inactivation of waterborne viruses to outperform the state-of-the-art ultrafiltration membranes.

Ace2 Substrate Cleavage Effect Associated with Covid-19 Patient Plasma Immunoglobulin

Background: Many mechanisms responsible for COVID-19 pathogenesis are well-known, but COVID-19 includes features with unclear pathogenesis, like autonomic dysregulation, coagulopathies, and high levels of inflammation. The SARS-CoV-2 spike protein receptor binding domain (RBD) receptor is angiotensin converting enzyme 2 (ACE2). We hypothesized that some COVID-19 patients may develop immunoglobulins (Igs) that have negative molecular image of RBD sufficiently similar to ACE2 to yield ACE2-like catalytic activity - ACE2-like 'abzymes'. Method(s): To explore this hypothesis, we studied 67 patients hospitalized with COVID-19 who had disodium ethylenediaminetetraacetate (EDTA) anticoagulated plasma samples available, obtained about 7 days after admission. We used commercially available fluorometric ACE2 assays (Abcam), and a SpectraMax M5 microplate reader (Molecular Devices), measuring Relative Fluorescent Unit (RFU, Ex/Em = 320/420 nm;RFU) in a kinetic mode every 20 min at 37C. ACE2 inhibitor provided in the assay kit was used for additional controls. In some control experiments, we added Zn2+ to plasma, or conducted serial dilutions to decrease Zn2+. To deplete Igs, we passed plasma samples through a 0.45 mum filter to remove large particles, then passed the material through 100kDa cut-off ultrafiltration membrane (PierceTM) columns, and finally used protein A/G Magnetic Beads (Life Technologies) to specifically deplete Ig, removing >99.99% of Ig as assessed with a human IgG ELISA Kit (Abcam). Result(s): ACE2 is a metalloprotease that requires Zn2+ for activity. However, we found that the plasma of 11 of the 67 patients could cleave a synthetic ACE2 peptide substrate, even though the plasma samples were collected using EDTA anticoagulant. When we spiked plasma with synthetic ACE2, no ACE2 substrate cleavage activity was observed unless Zn2+ was added, or the plasma was diluted to decrease EDTA concentration. After processing samples by size exclusion and protein A/G adsorption, the plasma samples did not cleave the ACE2 substrate peptide. Conclusion(s): The data suggest that some patients with COVID-19 develop Igs with activity capable of cleaving synthetic ACE2 substrate. Since abzymes can exhibit promiscuous substrate specificities compared to the enzyme whose active site image they resemble, and since proteolytic cascades regulate physiologic processes, anti-RBD abzymes may contribute to some otherwise obscure features of COVID-19 pathogenesis. (Figure Presented).

Evaluation of SARS-CoV-2 RNA Presence in Treated and Untreated Hospital Sewage.

Wastewater-based epidemiology (WBE) is a potential approach for determining the viral prevalence in a community. In the wake of the COVID-19 pandemic, researchers have begun to pay close attention to the presence of SARS-COV-2 RNA in various wastewaters. The potential for detecting SARS-CoV-2 RNA in hospital sewage could make it an invaluable resource for epidemiological studies. In this regard, two specialized hospitals dedicated to COVID-19 patients were chosen for this investigation. Both hospitals utilize the same wastewater treatment systems. The influent and effluents of the two hospitals were sampled in May and June of 2021, and the samples were evaluated for their chemical properties. According to the findings of this study, the wastewater qualities of the two studied hospitals were within the standard ranges. The sewage samples were concentrated using ultrafiltration and PEG precipitation techniques. The E and S genes were studied with RT-qPCR commercial kits. We found E gene of SARS-CoV-2 in 83.3% (5/6) and 66.6% (4/6) of wastewater samples from hospital 1 and hospital 2, respectively, using ultrafiltration concentration method. Wastewater samples taken after chlorine treatment accounted for 16.6% of all positive results. In addition, due to the small sample size, there was no significant correlation (p > 0.05) between the presence of SARS-CoV-2 in wastewater and the number of COVID-19 cases. Hospitals may be a source of SARS-CoV-2 pollution, thus it is important to monitor and enhance wastewater treatment systems to prevent the spread of the virus and safeguard the surrounding environment.

New generation washable PES membrane face mask for virus filtration

Membrane materials might be used for face protection because they can decontaminate the inhaled air from particle pollution and viruses like the SARS-Cov0-2 which damages our respiration system. In this study, plyethersulfone membranes (PES) were synthesized with green solvent at room temperature and its filtration effectiveness was investigated against nano-bacteria (size 0.05 to 0.2 µm) by measuring their Bacterial Filtration Efficiency (BFE) and micro aerosol size (0.3 µm), and Particulate Filtration Efficiency (PFE). The average SARS-CoV-2 diameters are between 50 nm to 160 nm. A series of experiments were performed to accomplish between 0.03 to 0.21 µm PES sponge like diameters so that can be used for SARS-CoV-2 filtration. Results showed that nanofiltration/ultrafiltration could filter 99.9% of bacteria and aerosol from contaminated air the size of the Covid-19 molecule.

Application of urinary proteomics for biomarker discovery in respiratory diseases

Background: To search for molecular biomarkers of pulmonary pathologies using non-invasive samples, such as urine, is of high clinical relevance. However, there are almost no proteomic studies using urine applied to respiratory diseases. Aim(s): To develop a biomarker discovery strategy using non-targeted proteomics in urine with applicability to different pulmonary diseases. Method(s): Urine samples were centrifuged and DTT treated to decrease uromodulin (THP). Low-THP samples were concentrated (ultrafiltration), ultracentrifugated, and exosome free urine was analysed using LC-MS/MS. GO terms/Pathway analyses were performed using STRING database. Result(s): Urine proteome (765 proteins) was enriched (FDR < 0.05) in proteins from different tissues, including respiratory system (N = 124), lung (N = 107), and immune system (N = 88). We detected an enrichment of relevant pathways for respiratory diseases, including several innate (e.g., TLR and NFkB pathways, complement system), and adaptive (e.g., interleukin signalling) immune system pathways. Some of these proteins have been previously studied in respiratory system disease (e.g., MPO, NAPSA, CHL1, FREM2, PLG), lower respiratory tract disease (e.g., NCAM1, MTOR, SERPINA1), viral infectious disease (e.g., ITIH4, CD209, CLEC4M, CD55), or specific pathologies such as coronavirus infection (e.g., ACE2, TMPRSS2), bronchiectasis (e.g., SAA1, SAA2, ELANE) or asthma (e.g., IGFALS, IGFBP7, HSPG2, DPP4, CD44, IL6R, MASP1). Conclusion(s): We have developed a protocol for the detection of proteomic biomarkers in urine. This proteome is enriched in proteins from the immune and respiratory systems, with a potential clinical and translational relevance.

The role of reactive nitrogen species as biomarkers of disease severity in COVID-19

Reactive nitrogen species (RNS) such as nitric oxide (NO) can be produced by local pulmonary cells and inflammatory cells, and they play a role in the pathogenesis of chronic pulmonary diseases and pulmonary infections. NO is an unstable compound turning to a more stable nitrite and nitrate rapidly. The objective of our study was to investigate the association between plasma nitrate+nitrite, nitrate, and nitrite levels and the severity of coronavirus disease (COVID)-19. Blood plasma samples of mild, moderate, and severe COVID-19 patients were collected from 2 different hospitals. Plasma of healthy subjects, who had never COVID-19 was used as controls (n=20 for each group). Samples were isolated by centrifugation following ultrafiltration prior to the commercial nitrate/nitrite colorimetric assay kit. Plasma nitrate+nitrite and nitrate levels, respectively, were significantly increased in severe patients (medians=34.4muM and 33.4muM), as compared to mild groups (medians=22.3muM and 20.6muM;p<0.05). In contrast, nitrite levels were significantly lower in severe patients (median= 0.8muM) than mild patients (median= 1.6muM;p<0.0001). Patients with severe disease were older (64.9 years) than the mild patients (50.6 years;p<0.05). In severe patients, age was positively correlated with nitrate+nitrite and nitrate levels, respectively (r=0.502, p=0.024;r=0.489, p=0.029). Our findings suggest that RNS may play a role in the pathogenesis of COVID-19 and that it may be considered as a marker of severity.

The relationship between dialysis adequacy and COVID-19 onset and mortality risk: Single center experience

Aim: We primarily aimed in this study was to evaluate risk factors for COVID-19 infection and if any association between dialysis inadequacy in COVID-19 infection in maintenance hemodialysis (MHD) patients. Secondly aimed to describe prevalence and risk factors associated with long-lasting symptoms of non-deceased COVID-19 MHD patients before vaccination. Material(s) and Method(s): One hundred one MHD patients infected with COVID-19 and 100 MHD patients without the infection were enrolled in this retrospective cross-sectional study. Risk factors for mortality, need to intensive care unit (ICU) stay and long-lasting symptoms were analyzed. Result(s): The mean age of patients was 59.13+/-13.58 years. COVID-19 infected patients had significantly higher number of patients with DM, COPD, CHF. The need for ICU was found to be statistically significantly higher in patients with COPD and DM. In our results, the patients who had lower Kt/V at admission hospital had more than 5 fold higher rate of COVID-19 those have higher Kt/V. We analyzed risk factors for mortality at, one year included higher age, higher CRP and lower base-line Kt/V were diagnostic criteria. Older MHD patients had a high frequent of long-lasting symptoms. Low Kt/V, low hemoglobin level and high CRP level associated with a higher risk of long-lasting symptoms (p=0.00, p=0.001, p=0.02) Discussion: We conclude that DM, CHF, COPD, older age, obesity were poor prognostic factors for in infected with COVID-19. Dialysis adequacy parameters of Kt/V, serum albumin level, hemoglobin level were significantly lower in need to ICU and deceased patients.Copyright © 2023, Derman Medical Publishing. All rights reserved.

MALIGNANT HYPERTENSION AFTER COVD-19 VACCINE INJECTION

Case;40 y/o male. Clinical course;The patient was transferred to our university hospital because of DOE and severe headache. He had been well and had no history of hypertension or obesity. He had experienced the COVID-19 vaccine injection two week before this visit. After the injection he had been experienced high fever and general fatigue as well as 7 kg of weight loss. On examnation, it was found that he had severe hypertension (190/110 mmHg) and hypertensive optic fundi. On chest X-ray, cardiomegaly and bilateral lung infiltrations was evident and biochemical data indicated renal dysfunction (serum creatinine 2.35 mg/dl), high levels of plasma renin activity (39.1 ng/ml/hour normal;0.6-3.9) and aldosterone concentration (176 pg/ml normal;4.0-82.1), and inflammatory changes (CRP = 23 mg/dl). We also found that increased levels of LDH and decreased levels of hemoglobin which indicated hemolytic anemia and thrombotic microangiopathy. After the control of high blood pressure by intravenous administration of Calcium channel blockades, We performed renal biopsy, which had a finding of diffuse findings of onion skin lesion and global glomerular sclerosis compatible with the diagnosis of malignant hypertension. Any secondary etiologies including renal artery disease or collagen disease had not been identified. Seven days after the admission, we started hemodialysis for this patient because of the renal failure was not resolved. We also had startred ACE inhibitors. We stopped the diuretics and minimized the ultrafiltration. Twenty-five days after the admission the patients was withdrawn from dialysis with the urine volume around 2000 ml/day and the serum creatinine concentration 5.29 mg/dl. He was discharged without any aid of dialysis and with small number of anti-hypertensives. Four months after the discharge, his serum creatinine concentration was 3.36 mg/dl and his blood pressure was 139/85 mmHg with the ACE inhibitor and calcium channel blockades. Conclusions;The case suggested that the malignant hypertension might be triggered by COVID-19 vaccine injection, which is of clinical importance.

Detection of human adenovirus F41 in wastewater and its relationship to clinical cases of acute hepatitis of unknown aetiology

As of 8 July 2022, the World Health Organization (WHO) have reported 1010 probable cases of acute hepatitis of unknown aetiology in children worldwide, including approximately 250 cases in the United Kingdom (UK). Clinical presentations have often been severe, with liver transplantation a frequent clinical outcome. Human adenovirus F41 (HAdV-F41) has been detected in most children with acute hepatitis, but its role in the pathogenesis of this infection has yet to be established. Wastewater-based epidemiology (WBE) has become a well-established tool for monitoring the community spread of SARS-CoV-2, as well as other pathogens and chemicals. In this study, we adopted a WBE approach to monitoring levels of HAdV-F40/41 in wastewater before and during an acute hepatitis outbreak in Northern Ireland. We report increasing detection of HAdV-F40/41 in wastewater, concomitant with increasing numbers of clinical cases. Amplicon whole genome sequencing further classified the wastewater-derived HAdV as belonging to the F41 genotype which in turn was homologous to clinically derived sequences. We propose that WBE has the potential to inform community surveillance of HAdV-F41 and can further contribute to the ongoing global discussion supporting HAdV-F41 involvement in acute hepatitis cases. © 2022 The Authors

Prevalence, clinical course and outcomes of COVID-19 in peritoneal dialysis (PD) patients: a single-center experience.

INTRODUCTION: There are limited data on the effects of COVID-19 on peritoneal dialysis (PD) patients. This study aimed to describe the impact of COVID-19 on the PD population. METHODS: A monocentric retrospective observational study was conducted on 146 consecutive PD patients followed from January 2020 to March 2022 at the University Hospital of Modena, Italy. RESULTS: Twenty-seven (18.4%) PD patients experienced 29 episodes of SARS-CoV-2 infection, corresponding to an incidence rate of 0.16 episodes/patient-year. Median age of COVID-19 patients was 60.4 (interquartile range [IQR] 50.2-66.5) years. In unvaccinated patients (n. 9), COVID-19 was always symptomatic and manifested with fever (100%) and cough (77.7%). COVID-19 caused hospital admission of three (33.3%) patients and two (22.2%) died of septic shock. COVID-19 was symptomatic in 83.3% of vaccinated subjects (n.18) and manifested with fever (61.1%) and cough (55.6%). Hospital admission occurred in 27.8% of the subjects but all were discharged home. Median SARS-CoV-2 shedding was 32 and 26 days in the unvaccinated and vaccinated groups, respectively. At the end of the follow-up, COVID-19 triggered the shift from PD to HD in two subjects without affecting the residual renal function of the remaining patients. Overall, COVID-19 caused an excess death of 22.2%. COVID-19 vaccination refusal accounted for only 1.6% in this cohort of patients. CONCLUSION: COVID-19 incident rate was 0.16 episodes/patient-year in the PD population. About one-third of the patients were hospitalized for severe infection. Fatal outcome occurred in two (7.4%) unvaccinated patients. A low vaccination refusal rate was observed in this population.

Filtration in Pharmaceutical Industries and Role of Textile

Filtration is considered the keystone for clarification and control of contamination in pharmaceutical and bi-opharmaceutical manufacturing. From production to in-process to chemical and research laboratories to the purification of water for sterile and nonsterile products, all of which involve some form of filtration in order to achieve a good manufacturing practice (GMP). Textile materials possess a significant contribution to the pharmaceutical filtration system. Textile material in pharmaceutical filtration is used in the form of filter media or medium. Flexible in nature, large pore distribution and non-metallic properties of textile materials have led to widespread use as filter media for many years. In filtration processes, a proper selection of filter media/ membrane material is usually the most critical aspect for ensuring efficient separation. Generally, solid-liquid and solid-gas separation is done by the filter media. This paper emphasises solid-liquid filtration. Moreover, this paper reviews the water requirement, filtration processes and the role of textile in the filtration system of pharmaceutical industries. This paper also offers insight into the current market trend and COVID-19 impact on the pharmaceutical filtration industry. Furthermore, gathered information may be helpful to those studying and working in pharmaceutical engineering, filtration technology, and wastewater treatment and can get knowledge about filtration systems.

Innovative treatment of wastewater containing of triclosan – SBR followed by ultrafiltration/adsorption/advanced oxidation processes

The COVID-19 pandemic has resulted in an increased use of disinfectants worldwide. One of the most popular active ingredients is triclosan (TRC), which is added to personal care products such as soap, wet wipes and dish soap. Negative aspect of TRC in wastewater is its release into the environment, since TRC is not completely removed in WWTPs. In the context of the rapidly increasing concentration of disinfectants in the wastewater influent, the aim of the study was to develop a method for treating wastewater containing TRC. Biological treatment in a sequential batch reactor (SBR) was used, and then the SBR effluent was treated in one of the tertiary processes i.e. ultrafiltration/adsorption/ozonation/ozonation+UV. In the ultrafiltration step, innovative membranes were used, such as membranes prepared from recycled polystyrene (rec-PS) or polyethersulfone mixed with single-walled carbon nanotubes (PES-SWCNT). Adsorption was conducted in fixed-bed columns packed with novel composite adsorbent containing halloysite and single-walled carbon nanotubes. TRC impaired the removal of phosphate and caused the degradation of activated sludge. Phosphate removal in the SBR-2 reactor (wastewater with TRC) decreased throughout the 6 weeks from 48.1 % to 38.1 %;in contrast, for SBR-1 (wastewater without TRC), it increased from 34.3 % to 71.8 %. The phosphate concentration of the effluent exceeded the permissible values, necessitating further treatment. Flocs from SBR-2 were smaller and looser compared to those from the control SBR. Biodegradation of TRC was noted in the 5th week of the operation as methyl TRC was detected in the effluent (0.07 mg/L). In ultrafiltration with PES-SWCNT membrane, phosphate, TRC, and methyl TRC were reduced by 33.7 %, 27.8 %, 36.8 %, respectively. On the other hand, adsorption, ozonation and ozonation+UV processes were highly suitable to treat SBR effluent. The phosphate, TRC and TRC byproducts removal degrees exceeded 90 % both for adsorption in the fixed-bed column as well as the ozonation and ozonation+UV processes.

Assessment of two volumetrically different concentration approaches to improve sensitivities for SARS-CoV-2 detection during wastewater monitoring.

Wastewater monitoring for severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2), the virus responsible for the global coronavirus disease 2019 (COVID-19) pandemic, has highlighted the need for methodologies capable of assessing viral prevalence during periods of low population infection. To address this need, two volumetrically different, methodologically similar concentration approaches were compared for their abilities to detect viral nucleic acid and infectious SARS-CoV-2 signal from primary influent samples. For Method 1, 2 L of SARS-CoV-2 seeded wastewater was evaluated using a dead-end hollow fiber ultrafilter (D-HFUF) for primary concentration, followed by the CP Select™ for secondary concentration. For Method 2, 100 mL of SARS-CoV-2 seeded wastewater was evaluated using the CP Select™ procedure. Following D-HFUF concentration (Method 1), significantly lower levels of infectious SARS-CoV-2 were lost (P value range: 0.0398-0.0027) compared to viral gene copy (GC) levels detected by the US Centers for Disease Control (CDC) N1 and N2 reverse-transcriptase quantitative polymerase chain reaction (RT-qPCR) assays. Subsamples at different steps in the concentration process were also taken to better characterize the losses of SARS-CoV-2 during the concentration process. During the centrifugation step (prior to CP Select™ concentration), significantly higher losses (P value range: 0.0003 to <0.0001) occurred for SARS-CoV-2 GC levels compared to infectious virus for Method 1, while between the methods, significantly higher infectious viral losses were observed for Method 2 (P = 0.0002). When analyzing overall recovery of endogenous SARS-CoV-2 in wastewater samples, application of Method 1 improved assay sensitivities (P = <0.0001) compared with Method 2; this was especially evident during periods of lower COVID-19 case rates within the sewershed. This study describes a method which can successfully concentrate infectious SARS-CoV-2 and viral RNA from wastewater. Moreover, we demonstrated that large volume wastewater concentration provides additional sensitivity needed to improve SARS-CoV-2 detection, especially during low levels of community disease prevalence.

Evaluation of a simple ultrafiltration method for concentration of infective canine parvovirus and feline coronavirus from cell culture supernatants.

Enrichment of viral infectious titers following its propagation by cell culture is desirable for various experimental studies. The performance of an ultrafiltration (UF) process to concentrate infectious titers of non-enveloped Canine parvovirus 2 (CPV-2) and enveloped Feline coronavirus (FCoV) obtained from cell culture supernatants was evaluated in this study, and compared with ultracentrifugation (UC) process. A mean gain of > 1.0 log10 TCID50/mL was obtained for CPV-2 with UF, which was comparable with the gain obtained by UC. On the other hand, the gain was lower (0.7-1.0 log10 TCID50/mL) for FCoV with UF in contrast to UC (> 2.0 log10 TCID50/mL). However, the lower retentate volume following UC (∼120 fold) compared to that following UF (∼10 fold) for either of the viruses suggests a trend of increased infectious titer retention in UF concentrates relative to UC concentrates. The simplistic UF process evaluated here thus has the potential for use in applications requiring increased infectious titers of CPV-2 and FCoV.

Society for the Advancement of Patient Blood Management 2022 Scientific Abstracts

The proceedings contain 46 papers. The topics discussed include: anemia assessment with benchmarking of red blood cell transfusion risk in cardiac surgery;clinical outcomes and therapeutic strategies for gastrointestinal bleeding in patients who decline transfusion;effect of patient blood management program on outcomes of the elderly with femur fracture who underwent orthopedic surgery;effect of ultrafiltration during cardiopulmonary bypass on viscoelastic profiles in cardiac surgery: retrospective analysis;hemoglobin based oxygen carrier treatment and clinical outcomes in severe anemia when blood is not an option;hemoglobin, lactate dehydrogenase, and FACIT-fatigue normalization in pegcetacoplan-treated patients with paroxysmal nocturnal hemoglobinuria;implementing three key blood management measures during COVID-19 related inventory shortages;and managing preoperative anemia using a novel algorithm to determine the preoperative target hemoglobin.

A Case of Acute Decompensated Heart Failure Complicated by Cardiorenal Syndrome Treated With Ultrafiltration.

The standard of care of treatment for acute decompensated heart failure (ADHF) complicated by cardiorenal syndrome (CRS) is diuretics. This case report is an example of how the institution of ultrafiltration early in the course of ADHF complicated by CRS for volume removal can be an alternative approach rather than escalation of the diuretic regimen when the initial diuretic regimen is ineffective and, in turn, yielding a good clinical course.

Fluid balance neutralization secured by hemodynamic monitoring versus protocolized standard of care in critically ill patients requiring continuous renal replacement therapy: study protocol of the GO NEUTRAL randomized controlled trial.

BACKGROUND: Fluid overload is associated with worse outcome in critically ill patients requiring continuous renal replacement therapy (CRRT). Net ultrafiltration (UFNET) allows precise control of the fluid removal but is frequently ceased due to hemodynamic instability episodes. However, approximately 50% of the hemodynamic instability episodes in ICU patients treated with CRRT are not associated with preload dependence (i.e., are not related to a decrease in cardiac preload), suggesting that volume removal is not responsible for these episodes of hemodynamic impairment. The use of advanced hemodynamic monitoring, comprising continuous cardiac output monitoring to repeatedly assess preload dependency, could allow securing UFNET to allow fluid balance control and prevent fluid overload. METHODS: The GO NEUTRAL trial is a multicenter, open-labeled, randomized, controlled, superiority trial with parallel groups and balanced randomization with a 1:1 ratio. The trial will enroll adult patients with acute circulatory failure treated with vasopressors and severe acute kidney injury requiring CRRT who already have been equipped with a continuous cardiac output monitoring device. After informed consent, patients will be randomized into two groups. The control group will receive protocolized fluid removal with an UFNET rate set to 0-25 ml h-1 between inclusion and H72 of inclusion. The intervention group will be treated with an UFNET rate set on the CRRT of at least 100 ml h-1 between inclusion and H72 of inclusion if hemodynamically tolerated based on a protocolized hemodynamic protocol aiming to adjust UFNET based on cardiac output, arterial lactate concentration, and preload dependence assessment by postural maneuvers, performed regularly during nursing rounds, and in case of a hemodynamic instability episode. The primary outcome of the study will be the cumulative fluid balance between inclusion and H72 of inclusion. Randomization will be generated using random block sizes and stratified based on fluid overload status at inclusion. The main outcome will be analyzed in the modified intention-to-treat population, defined as all alive patients at H72 of inclusion, based on their initial allocation group. DISCUSSION: We present in the present protocol all study procedures in regard to the achievement of the GO NEUTRAL trial, to prevent biased analysis of trial outcomes and improve the transparency of the trial result report. Enrollment of patients in the GO NEUTRAL trial has started on June 31, 2021, and is ongoing. TRIAL REGISTRATION: ClinicalTrials.gov NCT04801784. Registered on March 12, 2021, before the start of inclusion.

Unraveling the mystery of efficacy in Chinese medicine formula: New approaches and technologies for research on pharmacodynamic substances.

Traditional Chinese medicine (TCM) is the key to unlock treasures of Chinese civilization. TCM and its compound play a beneficial role in medical activities to cure diseases, especially in major public health events such as novel coronavirus epidemics across the globe. The chemical composition in Chinese medicine formula is complex and diverse, but their effective substances resemble "mystery boxes". Revealing their active ingredients and their mechanisms of action has become focal point and difficulty of research for herbalists. Although the existing research methods are numerous and constantly updated iteratively, there is remain a lack of prospective reviews. Hence, this paper provides a comprehensive account of existing new approaches and technologies based on previous studies with an in vitro to in vivo perspective. In addition, the bottlenecks of studies on Chinese medicine formula effective substances are also revealed. Especially, we look ahead to new perspectives, technologies and applications for its future development. This work reviews based on new perspectives to open horizons for the future research. Consequently, herbal compounding pharmaceutical substances study should carry on the essence of TCM while pursuing innovations in the field.

Potential Multifunctional Bioactive Compounds from Dysosma versipellis Explored by Bioaffinity Ultrafiltration-HPLC/MS with Topo I, Topo II, COX-2 and ACE2.

Background: Dysosma versipellis (D. versipellis) has been traditionally used as a folk medicine for ages. However, the specific phytochemicals responsible for their correlated anti-inflammatory, anti-proliferative and antiviral activities remain unknown. Purpose: This study aimed to explore the specific active components in D. versipellis responsible for its potential anti-inflammatory, anti-proliferative, and antiviral effects, and further elucidate the corresponding mechanisms of action. Methods: Bioaffinity ultrafiltration coupled to liquid chromatography-mass spectrometry (UF-LC/MS) was firstly hired to fast screen for the anti-inflammatory, anti-proliferative and antiviral compounds from rhizomes of D. versipellis, and then further validation was conducted using in vitro inhibition assays and molecular docking. Results: A total of 12, 12, 9 and 12 phytochemicals with considerable affinities to Topo I, Topo II, COX-2 and ACE2 were fished out, respectively. The anti-proliferative assay in vitro indicated that podophyllotoxin and quercetin exhibited comparably strong inhibitory rates on A549 and HT-29 cells compared with 5-FU and etoposide. Meanwhile, kaempferol displayed prominent dose-dependent inhibition against COX-2 with IC50 value at 0.36 ± 0.02 µM lower than indomethacin at 0.73 ± 0.07 µM. Furthermore, quercetin exerted stronger inhibitory effect against ACE2 with IC50 value at 104.79 ± 8.26 µM comparable to quercetin 3-O-glucoside at 135.25 ± 6.54 µM. Conclusion: We firstly showcased an experimental investigation on the correlations between bioactive phytochemicals of D. versipellis and their multiple drug targets reflecting its potential pharmacological activities, and further constructed a multi-target and multi-component network to decipher its empirical traditional applications. It could not only offer a reliable and valuable experimental basis to better comprehend the curative effects of D. versipellis but also provide more new insights and strategies for other traditional medicinal plants.