Clearance Method Research Articles

Determination of urine volume and glomerular filtration rate using d-serine and d-asparagine.

Measurement of glomerular filtration rate (GFR) is subject to inaccurate urine collection. Clearances of d-serine and d-asparagine, rare enantiomers of amino acids, are the measures of GFR since they are almost free of tubular secretion and reabsorption. We hypothesize that d-serine and d-asparagine can accurately determine urine volume and GFR. This cross-sectional observational study included 209 living kidney transplant donors and recipients for whom GFR was measured using the clearance of inulin. Assuming that urine excretions of d-serine and d-asparagine are constant and using urine levels of d-serine and d-asparagine from each urine collection, an equation for estimated urine volume (eUV) was established. Based on the eUV, the abnormal urine volume was replaced with an estimate with which the GFR was re-evaluated. Clearances of d-serine and d-asparagine were minor in proportional biases when compared with that of creatinine. Using 627 urine collection, the equation for eUV (mL/min) was established as 21.88/urine d-Ser (0.40+ 0.20 × log10(urine d-Asn)). Using eUV, we identified 20 instances where urine collection volumes varied significantly from the estimated values. After replacement with eUV, measured GFR (mGFR) was corrected to adjusted mGFR, which was within approximately 20mL/min/1.73m2 of the mGFR. d-Serine and d-asparagine are nearly completely excreted in urine after glomerular filtration, enabling the estimation of urine volume and correct mGFR. Besides reflecting GFR, d-serine and d-asparagine can be used to estimate urine volume. By applying the eUV method, mGFR determined using clearance methods becomes more accurate.

A review on the mucus dynamics in the human respiratory airway.

Research interest in the dynamics of respiratory flow and mucus has significantly increased in recent years with important contributions from various disciplines such as pulmonary and critical care medicine, surgery, physiology, environmental health sciences, biophysics, and engineering. Different areas of engineering, including mechanical, chemical, civil/environmental, aerospace, and biomedical engineering, have longstanding connections with respiratory research. This review draws on a wide range of scientific literature that reflects the diverse audience and interests in respiratory science. Its focus is on mucus dynamics in the respiratory airways, covering aspects such as mucins in fluidity and network formation, mucus production and function, response to external conditions, clearance methods, relationship with age, rheological properties, mucus surfactant, and mucoviscidosis. Each of these areas contains multiple subtopics that offer extensive depth and breadth for readers. We underscore the crucial importance of regulating and treating mucus for maintaining the health and functionality of the respiratory system, highlighting the ongoing need for further research to address respiratory disorders associated with mucus dynamics.

Revised Clinical Practice Guideline of Korean Pancreatobiliary Association for Common Bile Duct Stones: Endoscopic Management of Difficult Common Bile Duct Stones

Difficult common bile duct (CBD) stones are difficult to remove using conventional stone clearance methods such as endoscopic papillary sphincterotomy. In this guideline, we present several recommendations and relevant information on the endoscopic treatment of difficult CBD stones. For the primary treatment of large CBD stones in patients without distal bile duct stricture, we recommend limited endoscopic sphincterotomy followed by endoscopic papillary large balloon dilation. For large CBD stones that are difficult to remove with conventional endoscopic treatment methods, we recommend electrohydraulic or laser lithotripsy under cholangioscopy. Short-term biliary stenting is recommended in patients with CBD stones that are difficult to remove by these methods. Finally, endoscopic treatment via small bowel enteroscopy or percutaneous transhepatic cholangioscopy may be an option in patients with surgically altered anatomy.

Immunomodulatory Effects of Citrus Species Ethanol Extracts on Cellular Immune Responses in Wistar Rats and Mice

Currenty, Citrus reticulata has been found to modulate RAW 264.7 macrophage cell line. In the present study, the immunomodulatory effects of Citrus species peel extracts, including Citrus microcarpa, C. limon L., C. sinensis L. and C. hystrix were investigated on cellular immune responses in Wistar rats and mice. Mice were treated with ethanol extracts of Citrus species orally at dosages of 25, 50 and 100 mg/kg bw for 7 days. The phagocytosis activity of the extracts was examined using the carbon clearance method. Meanwhile, oral administration of Citrus species ethanol extracts at dosages of 17.5, 35 and 70 mg/kg bw was administered to Wistar rats to assess the effect of extracts on the delayed-type hypersensitivity (DTH) response using paw edema-method. Thin Layer Chromatography (TLC)-densitometry analysis was also performed. In both animal models, all extracts enhanced the cellular immune responses. Amongst the samples, C. sinensis, at the dose of 100 mg/kg bw in mice and 70 mg/kb bw in rats, demonstrated the highest stimulation on phagocytosis activity and DTH response, respectively. Except C. microcarpa, rutin was found in all Citrus species. The results indicate that Citrus species, particularly C. sinensis, have the potential to be developed into a potent immunostimulatory agent. HIGHLIGHTS The immunomodulatory effects Citrus species were investigated using mice and rats as animal models. Citrus microcarpa, C. limon, C. sinensis L. and C. hystrix were able to stimulate phagocytosis in mice and delayed-type hypersensitivity response in rats. Citrus sinensis demonstrated the highest stimulation on phagocytosis activity and delayed-type hypersensitivity response. Rutin was found in all Citrus species, except in microcarpa peels extract. GRAPHICAL ABSTRACT

Antioxidant and immunomodulatory activities of ethanol extracts from Syzygium cumini L. Skeels and Pogostemon cablin Benth

Syzygium cumini and Pogostemon cablin are mostly cultivated in tropical climates for culinary and perfumery purposes, yet their potential medicinal properties remain underreported. The aim of this study was to examine the antioxidant and immunomodulatory activities of ethanol extracts from S. cumini (EESC) and P. cablin (EEPC). Reflux extraction was carried out using 96% ethanol on the collected plant specimens to produce EESC and EEPC. Secondary metabolites of each extract were identified using gas chromatography-mass spectrometry (GC-MS). The extracts were measured for total phenol and flavonoid levels and 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity. The immunomodulatory activity test was carried out in vivo by assessing several parameters, including the phagocytic index via the carbon clearance method, organ indices, antibody titers, and delayed-type hypersensitivity (DTH) response, using sheep red blood cells (SRBC) as antigens. The extracts were also examined for their anti-inflammatory activity in acute and chronic inflammation models. In the DPPH antioxidant test, EESC and EEPC had IC50 values of 12.33 µg/mL and 182.17 µg/mL, respectively. Both extracts showed immunosuppressant activity, marked by a phagocytic index of <1. EESC yielded lower organ indices for the liver (p=0.025 at 200 mg/kg BW), spleen (p=0.028 at 100 mg/kg BW), and thymus (p=0.032 at 200 mg/kg BW) compared to the control group. For EEPC, lower organ indices were observed in the liver at 100 mg/kg BW (p=0.005) and 200 mg/kg BW (p=0.031). In the primary antibody titer and DTH tests, both EESC and EEPC showed immunosuppressant activity at 200 and 400 mg/kg BW (p<0.05). The extracts suppressed both the innate and adaptive immune systems. Both EEPC (p=0.004) and EESC at 100 mg/kg BW (p=0.03) significantly reduced serum TNF-α levels. In conclusion, EESC and EEPC have the potential as immunosuppressive and anti-inflammatory agents.

Glomerular filtration rate determined by measuring serum clearance of iohexol in unanesthetized cheetahs (Acinonyx jubatus) with comparison to serum symmetric dimethylarginine.

One of the more common diseases affecting zoo-managed cheetahs (Acinonyx jubatus) is chronic renal disease, which can impact their welfare and ultimately shortens their lifespan. Early diagnosis, for which estimating Glomerular Filtration Rate (GFR) is one such tool, is imperative to help mitigate the negative impacts of this insidious disease. GFR was determined by measuring the serum clearance of iohexol in nine clinically normal, cheetahs managed under human care that presented for voluntary blood collection. A 2-sample iohexol clearance method was performed, along with serum symmetric dimethylarginine (SDMA) determination. SDMA has shown promise in humans, dogs, and cats, as an early biomarker of renal disease. Additionally, the relationship between GFR and SDMA, along with serum creatinine and BUN were analyzed. The mean values for the uncorrected GFR and corrected GFR were 2.08 ± 0.215 mL/min/kg body weight and 1.87 ± 0.173 mL/min/kg body weight, respectively. No significant correlations were observed between GFR, SDMA, serum creatinine, or BUN. Both the uncorrected and corrected iohexol-based GFR values were similar to an inulin-based GFR reference interval determined in zoo managed cheetahs and a reported domestic cat iohexol-based GFR reference interval. Serum SDMA values support previous research suggesting cheetahs have a separate reference interval from domestic cats (0-14 μg/dL). Measuring GFR by the serum clearance of iohexol shows promise as a readily available, cheap, and easily administered clearance marker that can be used in cheetahs trained for voluntary blood collection, thereby avoiding the need for anesthesia.

Antioxidant, Antibacterial and Immunomodulatory Properties of Algerian Phoenix dactylifera

The aim of the current study is to evaluate the antioxidant, antibacterial, and phagocytic activities of the hydroethanolic extract of Phoenix dactylifera fruit variety Mech degla. The antioxidant activity was determined through DPPH assay, ferrous ions chelating assay, and CUPRAC assay. The antibacterial activity against four bacteria (Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus pneumoniae), was evaluated using the disc diffusion method, while the in vivo phagocytic activity was measured using carbon clearance method. The results showed that the extract exhibited good antioxidant activity with the IC50 values of 0.89 ± 0.3 g/l and 0.56 ± 0.24 g/l for DPPH and ferrous ions chelating activities, respectively, and the A0.5 value of 1.45 ± 0.15 g/l in the CUPRAC assay. Moreover, the extract showed notable antibacterial activity against all tested strains with the highest zone of inhibition (23.3 ± 0.3 mm) against S. pneumonia. Besides the extract did not show any significant toxicity, it was able to stimulate the phagocytic activity with a significantly faster carbon clearance rate at the dose of up to 100 mg/kg compared to the control group. Our results suggested that the Phoenix dactylifera fruit represents a promising source for the development of safe and effective immunomodulators as remedies to prevent and treat various microbial and immune-related diseases.

Abstract P178: Sex Differences in the Role of the Immune ET-1/ET A Axis in Controlling Blood Pressure during a Hypertension Challenge

Immune cells like dendritic cells (DCs) and T cells are essential for the development of hypertension. Upregulation of the vasoactive peptide endothelin-1 (ET-1) is key in the progression of hypertension. ET-1 acting via its ET A receptor raises blood pressure and promotes end-organ damage and inflammation. DCs and T cells express ET-1 receptors and respond to changing levels of ET-1. However, it is unknown whether activation of the ET-1/ET A receptor axis on DCs or T cells is critical in driving hypertension, or if a differential activation of this axis in immune cells leads to the sex differences that exist in the pathophysiology of this disease. We hypothesized that activation of the ET-1/ ET A axis on immune cells is crucial in the development of hypertension, with a more prominent role in males than females. Male and female mice lacking ET A receptor on DCs (DC ET A KO) or on T cells (T cell ET A KO) and floxed ET A control mice were given the nitric oxide synthase inhibitor L-NAME (0.5 mg/mL) in drinking water for 3 wks to induce hypertension. Systolic blood pressure (SBP) was measured 3 times/wk via tail cuff plethysmography. Glomerular filtration rate (GFR) was evaluated at baseline and after L-NAME treatment via the transcutaneous FITC-sinistrin clearance method. Absence of ET A receptor on DCs resulted in a ~10 mmHg decrease in SBP at baseline in both sexes, and this difference increased throughout the protocol in males (Floxed ET A vs. DC ET A KO: L-NAME: 150.8±6.7 vs. 130.8±2.6 mmHg, n=4/group, p=0.002), but not in females (137.4±5.7 vs. 137.2±2.4 mmHg, n=3/group). Lack of the ET A receptor on T cells led to a similar decrease of ~10 mmHg in SBP at baseline. However, the difference in SBP widened when males were challenged with L-NAME (Floxed ET A vs. T cell ET A KO: 135.2±2.0 vs. 118.0±4.1, n=11/group, p=0.003), whereas the group difference in SBP disappeared in females (131.5±4.0 vs. 132.4±4.7, n=8-9/group). Following L-NAME, GFR trended to decline in control male mice, while it increased in T cell ET A KO males. Our data suggest that activation of ET A receptors on DCs and T cells modulates SBP in response to a hypertensive insult in males, but not in females. Additionally, absence of ET-1/ ET A axis activation on T cells suggests improved renal function during hypertension. Our results suggest that the use of ET A antagonism in the clinic may be more protective during hypertension in males than females.

The safety of not implementing endoscopic nasobiliary drainage after elective clearance of choledocholithiasis: a systematic review and meta-analysis

BackgroundEndoscopic nasobiliary drainage (ENBD) is used as a drainage technique in patients with choledocholithiasis after stone removal. However, ENBD can cause discomfort, displacement, and other complications. This study aims to evaluate the safety of not using ENBD following elective clearance of choledocholithiasis.MethodsRelevant studies were identified by searching PubMed, Web of Science, EMBASE, EBSCO, and Cochrane Library from their inception until August 2023. The main outcomes assessed were postoperative complications and postoperative outcomes. Subgroup analyses were conducted based on study design types and treatment procedures.ResultsSix studies, including three randomized controlled trials (RCTs) and three cohort studies, were analyzed. Among these, four studies utilized endoscopic techniques, and two employed surgical methods for choledocholithiasis clearance. The statistical analysis showed no significant difference in postoperative complications between the no-ENBD and ENBD groups, including pancreatitis (RR: 1.55, p = 0.36), cholangitis (RR: 1.81, p = 0.09), and overall complications (RR: 1.25, p = 0.38). Regarding postoperative outcomes, the subgroup analysis indicated that the bilirubin normalization time was longer in the no-ENBD group compared to the ENBD group in RCTs (WMD: 0.24, p = 0.07) and endoscopy studies (WMD: 0.23, p = 0.005), although the former did not reach statistical difference. There was also no significant difference in the length of postoperative hospital stay between the groups (WMD: -0.30, p = 0.60).ConclusionIt appears safe to no- ENBD after elective clearance of choledocholithiasis.

Extracellular Vesicles in Ovarian Cancer: From Chemoresistance Mediators to Therapeutic Vectors.

Ovarian cancer (OC) remains the deadliest gynecological malignancy, with alarming projections indicating a 42% increase in new cases and a 51% rise in mortality by 2040. This review explores the challenges in OC treatment, focusing on chemoresistance mechanisms and the potential of extracellular vesicles (EVs) as drug delivery agents. Despite advancements in treatment strategies, including cytoreductive surgery, platinum-based chemotherapy, and targeted therapies, the high recurrence rate underscores the need for innovative approaches. Key resistance mechanisms include drug efflux, apoptosis disruption, enhanced DNA repair, cancer stem cells, immune evasion, and the complex tumor microenvironment. Cancer-associated fibroblasts and extracellular vesicles play crucial roles in modulating the tumor microenvironment and facilitating chemoresistance. EVs, naturally occurring nanovesicles, emerge as promising drug carriers due to their low toxicity, high biocompatibility, and inherent targeting capabilities. They have shown potential in delivering chemotherapeutics like doxorubicin, cisplatin, and paclitaxel, as well as natural compounds such as curcumin and berry anthocyanidins, enhancing therapeutic efficacy while reducing systemic toxicity in OC models. However, challenges such as low production yields, heterogeneity, rapid clearance, and inefficient drug loading methods need to be addressed for clinical application. Ongoing research aims to optimize EV production, loading efficiency, and targeting, paving the way for novel and more effective therapeutic strategies in OC treatment. Overcoming these obstacles is crucial to unlocking the full potential of EV-based therapies and improving outcomes for OC patients.

GFR measurement in patients with CKD: Performance and feasibility of simplified iohexol plasma clearance techniques.

Implementing shortened one-compartment iohexol plasma clearance models for GFR measurement is crucial since the gold standard inulin renal clearance technique and the reference two-compartment, 10-hour, 16-samplings iohexol plasma clearance method are clinically unfeasible. Inulin may precipitate anaphylactic shock. Four-hour and 8-hour one-compartment iohexol plasma clearance models with Bröchner-Mortensen correction provide accurate GFR measurements in patients with estimated GFR (eGFR) > or ≤40 mL/min/1.73m2, respectively. We compared the performance of the simplified 5-hour, 4-samplings, two-compartment population pharmacokinetic model (popPK) with the performance of the reference two-compartment 10-hour iohexol method in 16 patients with GFR 15.2 to 56.5 mL/min/1.73 m2. We also compared the performance of shortened (5, 6 and 7-hour) one-compartment models with the performance of the standard 8-hour one-compartment model in 101 patients with eGFR ≤40 mL/min/1.73 m2. The performance of popPK and shortened methods versus reference methods was evaluated by total deviation index (TDI), concordance correlation coefficient (CCC) and coverage probability (CP). TDI <10%, CCC ≥0.9 and CP >90% indicated adequate performance. TDI, CCC and CP of popPK were 11.11%, 0.809 and 54.10%, respectively. All shortened, one-compartment models overestimated the GFR (p <0.0001 for all) as compared to the 8-hour model. TDI, CCC and CP were 7.02%, 0.815, and 75.80% for the 7-hour model, 7.26%, 0.803, and 74.20% for the 6-hour model, and 8.85%, 0.729 and 64.70% for the 5-hour model. The agreement of popPK model was comparable to that obtained with the Chronic-Kidney-Disease-Collaboration-Epidemiology (CKD-Epi) and the Modification-of-Diet-in-Renal-Disease (MDRD) serum-creatinine based equations for GFR estimation. PopPK model is remarkably unreliable for GFR measurement in stage III-IV CKD patients. In patients with eGFR ≤40 mL/min/1.73m2, shortened one-compartment models, in particular the 5-hour model, are less performant than the reference 8-hour model. For accurate GFR measurements, the iohexol plasma clearance should be measured with appropriate protocols. Over-simplified procedures should be avoided.

Comparing effects of inspiratory muscle trainer and lung flute on sputum clearance in chronic obstructive pulmonary disease: A randomized controlled trial

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a preventable and treatable disease. Mucus accumulation, a common impairment in COPD affects lung function, exacerbations and quality of life. Physiotherapy measures including positive expiratory pressure devices like lung flute form the non-pharmacologic treatment options for the same. Although not a primary airway clearance device, inspiratory muscle trainer (IMT) plays a role in it. The aim of this study was to compare the effectiveness of lung flute and IMT as methods for sputum clearance in COPD. METHOD: Participants with COPD were recruited from the Department of Pulmonology and screened for eligibility. Block randomization was used to divide the participants into three groups: Inspiratory muscle training (IMT), lung flute (LF), and routine physiotherapy (RP). Supervised 30 minutes of in-patient intervention was delivered to all the groups for 6 days, once a day. Total sputum volume analysis (TSV) was conducted before and after 6 days of intervention. Descriptive statistics were presented as median and quartiles. Kruskal Wallis H test and Wilcoxon signed rank test were applied. RESULTS: 60 participants aged 55 to 65 years with mild to moderate COPD were included in the study. Median TSV for RP, LF and IMT were 8.50, 20.50 and 25.75 ml respectively. Pair-wised comparisons for TSV revealed significant differences for the means of the rank scores between RP and LF (LF dominant) (p &lt; 0.001), RP and IMT (IMT dominant) (p &lt; 0.001) and LF and IMT (IMT dominant) (p = 0.002). CONCLUSION: IMT performed superiorly in terms of sputum clearance in patients with COPD.

Percutaneous nephrolithotomy for adult renal transplant de novo graft lithiasis: a single center analysis and systematic review of the literature.

Graft stones in renal transplant recipients pose a unique challenge, finding effective interventions to ensure optimal graft function and patient well-being. Various methods of stone clearance have been described for graft stones, including percutaneous nephrolithotomy (PCNL). While PCNL is a promising approach for managing graft stones, specific outcomes and associated characteristics for this approach have not been comprehensively evaluated before. This study aims to evaluate the safety and efficacy of the use of PCNL as the primary intervention of graft stones by assessing stone-free rates (SFR), treatment impact on graft function, and perioperative complications. A retrospective clinical audit was performed for all transplants performed in a single center from 2007 to 2022, which included all graft lithiasis patients who were treated with PCNL. Both perioperative parameters and post-operative outcomes were collected. In addition, a systematic review including articles from MEDLINE, Embase, Web of Science yielded 18 full-text articles published between 1/1/2000 and 15/11/2023. The results pertaining to patients who underwent PCNLs for graft stones were cross-referenced and thoroughly evaluated. The review encompassed a comprehensive analysis of clinical data, postoperative outcomes, and procedural details. The protocol for the systematic review was prospectively registered on PROSPERO (CRD42023486825). In our center, 6 graft lithiasis patients were treated with PCNL. The initial SFR was 83.3%. SFR at 3months and 1year were both 100.0%. SFR at 3years was 66.7%. Other centers reported initial SFR of 82.6-100.0% (interquartile range). SFR at 3months, 1year, 3years was not well reported across the included studies. Incidence of graft lithiasis ranged from 0.44%-2.41%. Most common presentations at diagnosis were oliguria/anuria/acute kidney injury and asymptomatic. Reported complications included blood loss, transient hematuria, high urine output, sepsis, and damage to surrounding structures. The most commonly reported metabolic abnormalities in transplant lithiasis patients included hyperuricemia and hyperparathyroidism. PCNL is a practical and efficient choice for addressing graft lithiasis, demonstrating excellent stone clearance and minimal perioperative complications. These findings show the importance of PCNL as a primary intervention in this complex patient population.

99m Tc-DTPA dynamic SPECT/CT renogram in adults: feasibility and diagnostic benefit.

The objective of this study is to investigate the feasibility and potential advantages of 99m Tc-DTPA dynamic single photon emission computed tomography/computed tomography (SPECT/CT) renogram in adults. Fifty-five patients aged 19-80 years (mean 56.3) were enrolled. The imaging protocol included: day 1: 99m Tc-DTPA planar renogram, followed by planar 99m Tc-DMSA scan. Day 3: attenuation-corrected dynamic 99m Tc-DTPA SPECT renogram [DSPECT(AC)] and Cr-51 ethylenediamine tetraacetic acid (EDTA) glomerular filtration rate (GFR) calculation. DSPECT(AC) included an initial CT scan followed by 12 consecutive SPECT sessions acquired via continuous-mode acquisition for a total of 24 min. Fast SPECT sequences (1-2 s/projection, 60 projections, every 6°) were obtained for the first 8 min, followed by slower acquisitions (3-4 s/projection) during the rest of the study. Renal activity was measured in the total kidney volume by regions of interest drawn on consecutive transaxial slices of the third SPECT, which were then copied on the whole 12-SPECT series. Corresponding time-activity curves were created. DSPECT(AC) parameters were compared with those of planar renogram. The reference method for split renal function was 99m Tc-DMSA (geometrical mean of anterior and posterior projection counts) and for GFR the Cr-51 EDTA 2-blood sample clearance method. DSPECT(AC) images were of good quality. There was good correlation between renogram parameters (time to peak activity and NORA20) comparing the two techniques ( r = 0.959 and 0.933, respectively). In 21 cases with >30% absolute difference between the two kidneys, spilt renal function calculation by DSPECT(AC) correlated perfectly ( r = 0.968) with the reference method, whereas planar renogram was less accurate ( r = 0.843). Anatomic information provided by nonenhanced CT offered an integrated structural-functional view valuable for final diagnosis. DSPECT(AC) early kidney uptake as a fraction of injected dose correlated better with reference GFR ( r = 0.789) than the Gates' method ( r = 0.642). 99m Tc-DTPA dynamic SPECT/CT renogram is feasible with conventional SPECT/CT systems. It allows accurate split renal function measurement, offers additional anatomical information and can be used for closer approximation of GFR compared with Gates' method.

Fate control engagement augments NK cell responses in LV/hu-IL-12 transduced sarcoma

IntroductionNK cells are an untapped resource for cancer therapy. Sarcomas transduced with lentiviruses to express human IL-12 are only cleared in mice bearing mature human NK cells. However, systemic inflammation limits IL-12 utilization. Fate control a.k.a. “suicide mechanisms” regulate unchecked systemic inflammation caused by cellular immunotherapies. Despite increasing utilization, there remains limited data on immune consequences or tumor-directed effects of fate control. ObjectivesWe sought to engage the mutant thymidylate kinase (mTMPK) metabolic fate control system to regulate systemic inflammation and assess the impact on NK cell effector functions. MethodsPrimary human sarcoma short-passage samples and cell lines were transduced with LV/hu-IL-12_mTMPK engineering expression of IL-12 and an AZT-associated fate control enzyme. We assessed transduced sarcoma responses to AZT engagement and subsequent modulation of NK cell functions as measured by inflammatory cytokine production and cytotoxicity. ResultsAZT administration to transduced (LV/hu-IL-12_mTMPK) short-passage primary human sarcomas and human Ewing sarcoma, osteosarcoma, and rhabdomyosarcoma cell lines, abrogated the robust expression of human IL-12. Fate control activation elicited a specific dose-dependent cytotoxic effect measured by metabolic activity (WST-1) and cell death (Incucyte). NK effector functions of IFN-γ and cytotoxic granule release were significantly augmented despite IL-12 abrogation. This correlated with preferentially induced expression of NK cell activation ligands. ConclusionsmTMPK fate control engagement terminates transduced sarcoma IL-12 production and triggers cell death, but also augments an NK cell-mediated response coinciding with metabolic stress activating surface ligand induction. Fate control engagement could offer a novel immune activation method for NK cell-mediated cancer clearance.

Identification of Bacterial Species Capable of Degrading LDPE

Abstract: The Plastic is polymeric material having capability of being molded or shaped by the application of heat and pressure. The low-density polyethylene(LDPE) is the type of plastic which is used in various purpose of our daily life. It is one of the major environmental problems because of its slow degradation rate .To control the pollution caused by low-density polyethylene, microbes are isolated from the dumping site of the Durg region of Chhattisgarh. These microbes are screened by the zone of clearance method for degradation of low-density polyethylene using PEG as substrate. The screened microbes were identified by morphological and biochemical characteristics.Nine bacterial isolates was selected for biodegradation studies.Confirmation of bacterial biodegradation of low-density polyethylene was performed by using weight loss measurement and pH change due to production of acidic product.One potential isolate were further identified through 16S rRNA sequences Alcaligenes faecalis was shown the high degradation rate of low-density polyethylene

NURSING CARE ANALYSIS OF ARTIFICIAL RESPIRATORY MANAGEMENT IN STROKE-ASSOCIATED PNEUMONIA (SAP) PATIENTS: A CASE STUDY FROM ICU RSPAL DR. RAMELAN SURABAYA

Introduction: Stroke-associated pneumonia (SAP) is a common complication of stroke, leading to ineffective airway clearance and potentially severe outcomes such as respiratory failure and death. Objective: This case study analyzes the nursing care provided to a SAP patient in the ICU of RSPAL Dr. Ramelan Surabaya, focusing on the application of artificial respiratory management to improve airway clearance Methods: Over five days, the nursing team implemented interventions, including Endotracheal Tube (ETT) care, which partially resolved the ineffective airway clearance. Results: This improvement was evident in the enhanced respiratory parameters and reduced secretion accumulation. Conclusions: The findings highlight the importance of diligent airway management in preventing respiratory complications in SAP patients.

Evaluation of Various Approaches to Estimate Transplacental Clearance of Vancomycin for Predicting Fetal Concentrations using a Maternal-Fetal Physiologically Based Pharmacokinetic Model.

Evaluating drug transplacental clearance is vital for forecasting fetal drug exposure. Ex vivo human placenta perfusion experiments are the most suitable approach for this assessment. Various in silico methods are also proposed. This study aims to compare these prediction methods for drug transplacental clearance, focusing on the large molecular weight drug vancomycin (1449.3g/mol), using maternal-fetal physiologically based pharmacokinetic (m-f PBPK) modeling. Ex vivo human placenta perfusion experiments, in silico approaches using intestinal permeability as a substitute (quantitative structure property relationship (QSPR) model and Caco-2 permeability in vitro-in vivo correlation model) and midazolam calibration model with Caco-2 scaling were assessed for determining the transplacental clearance (CLPD) of vancomycin. The m-f PBPK model was developed stepwise using Simcyp, incorporating the determined CLPD values as a crucial input parameter for transplacental kinetics. The developed PBPK model of vancomycin for non-pregnant adults demonstrated excellent predictive performance. By incorporating the CLPD parameterization derived from ex vivo human placenta perfusion experiments, the extrapolated m-f PBPK model consistently predicted maternal and fetal concentrations of vancomycin across diverse doses and distinct gestational ages. However, when the CLPD parameter was derived from alternative prediction methods, none of the extrapolated maternal-fetal PBPK models produced fetal predictions in line with the observed data. Our study showcased that combination of ex vivo human placenta perfusion experiments and m-f PBPK model has the capability to predict fetal exposure for the large molecular weight drug vancomycin, whereas other in silico approaches failed to achieve the same level of accuracy.

Pharmacokinetic parameters of D-glucitol in laboratory animals in the context of assessing the functional state of the liver

One of the most developing methods for assessing the state of the hepatobiliary system is the dynamic clearance method (assessment of clearance by studying pharmacokinetic parameters). Clearance is a measure of the rate at which a specific (usually most pharmacodynamically related to the system being studied) substance is eliminated from the body. This method allows not only to diagnose liver diseases, but also to determine their severity and the dynamics of changes in the process of further pharmacotherapy. The purpose of this study is to conduct an experimental study aimed at studying changes in the pharmacokinetics of exogenous sorbitol in the blood plasma of laboratory animals, which will establish starting points for further assessment of changes in its elimination properties in hepatobiliary pathologies. The level of Dglucitol was determined by the Corcoran and Page method with modification for 12 hours (step - 2 hours) in blood serum obtained by venipuncture of the tail vein, after administration of an experimental composition based on D-glucitol. Time intervals were selected based on the reported half-life of the drug. Based on the results of the study, we noted the following patterns in the biotransformation of D-glucitol: a linear increase in concentration up to 6 hours after administration (with a peak at 6 hours), a linear decrease in the period from 6 to 8 hours after administration, then a sharp decrease in concentration. Thus, the type of elimination of D-glucitol in clinically healthy animals can be assessed as “linear”. In the process of further research, it is planned to develop special “D-glucitol elimination curves”. These graphs will show the dynamics of exogenous D-glucitol removal over time in various hepatobiliary disorders.

Abstract 2061: A proven activity-based workflow for the identification and characterization of time-dependent kinase inhibitors using a continuous assay format

Abstract Time-dependent inhibitors (TDIs) of enzyme targets offer distinct advantages for the development of potent and selective compounds with favorable pharmacokinetic and pharmacodynamic properties. Such inhibitors are characterized by non-linear progress curves: after an initial inhibited velocity, a rate constant governs the transition to a final steady-state reaction rate of the inhibited enzyme. A final rate of zero indicates irreversible inhibition, whereas a non-zero final rate indicates slow-binding inhibition. Characterizing these inhibitory modes of action is enabled with a continuous assay format that avoids the common pitfalls and misleading results seen with end-point assays. A continuous assay format enables efficient and robust determination of the kinetic parameters required to drive structure-activity relationship optimization to streamline the development of more effective drugs. It is important to note that simple IC50s for TDIs will not suffice, and can, indeed, also be misleading. We have developed a robust three-step workflow based on kinetic catalytic activity measurements to quickly identify and characterize TDIs. First, dose-response experiments are conducted with and without an enzyme-inhibitor preincubation step. The curvature of the reaction progress curve in the non-preincubated experiment and a shift in IC50 from the preincubated experiment are indicative of TDI. In the absence of TDI, simple IC50s are reported with, if possible, Ki values. If TDI is present, a second experiment is conducted to assess compound reversibility using either a jump-dilution protocol or a novel free-compound clearance method that uses gel filtration spin columns or spin plates. In either protocol, forward progress curve analysis is used to monitor the recovery of enzymatic activity after dilution of inhibitor in solution. Lastly, the potency of the inhibitor is evaluated using kinetic experiments tailored to the nature of the inhibition – either reversible or irreversible. If reversible, then the rate constant from the reversibility experiment is used to determine the residence time of the molecule. If irreversible, then a 24-point dose-response experiment with serial 1.5-fold dilutions is performed, and all the progress curves are globally fit to determine kinact/KI, and, if possible, kinact and KI separately. The method will be fully described through the characterization of known EGFR inhibitors of three inhibition types: fast-off (Gefitinib), slow binding (Lapatinib), and irreversible (Osimertinib). Citation Format: Earl May, Daniel Urul, Khanh Huynh, Susan Cornell-Kennon, Venkatesh Nemmara, Zhibing Lu, Samuel Hoare, Michelle Lyles, Erik Schaefer. A proven activity-based workflow for the identification and characterization of time-dependent kinase inhibitors using a continuous assay format [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2061.