Toxic Injury Research Articles

Long-term complications and health-related quality of life outcomes after radical prostatectomy with or without subsequent radiation treatment for prostate cancer

BackgroundTo report objective long-term complications and health related quality of life (HRQOL) outcomes after radical prostatectomy (RP) with and without radiation therapy (RT) for prostate cancer (CaP). MethodsWe analyzed patients diagnosed with CaP who underwent RP from the UCSF Cancer of the Prostate Strategic Urologic Research Endeavor (CaPSURE) registry between 1995 and 2020. Cox proportional hazards were used to assess risk of postoperative complications which included cystitis, gastrointestinal (GI) toxicity, incontinence requiring a surgical procedure, ureteral injury and urinary stricture. Repeated measures mixed models were used to assess the effects of radiation and complications on patient-reported urinary, bowel, and sexual function after surgery. ResultsOf 6,258 men who underwent RP, cumulative incidence of EBRT was 9.1% at 5 years after surgery. Patients who received postoperative radiation were at increased risk for onset of cystitis (HR 5.60, 95% CI 3.40–9.22, P < 0.01). Receipt of RT was not associated with other complications. In repeated measures analysis, postoperative RT was associated with worsening general health scores, adjusting for complications of incontinence, urinary stricture, GI toxicity or ureteral injury, independent of whether patients had those complications. ConclusionsRT after RP was associated with an increase in the risk of cystitis and worse general health in the long term. Other complications and HRQOL outcomes did not demonstrate differences by whether patients had RT or not. While post-operative RT is the only curative option for CaP after RP, patients and providers should be aware of the increased risks when making treatment decisions.

An Analysis of Domestic Fire Smoke/Toxic Fumes Inhalation Injuries

Previous studies indicated that inhalation of smoke and toxic fumes is the most common form of domestic fire injury. In this article we examine the socio-demographic, contributory, and firefighting behaviour factors associated with accidental dwelling fire smoke / toxic fumes inhalation injuries. In particular, we examine age band, gender, occupancy level, deprivation, alcohol / drug consumption, and attempting to fight a domestic fire in the Merseyside area of North-West England covered by Merseyside Fire and Rescue Service over the period 2011 to 2022. Smoke / toxic fumes inhalation injuries occurred more in single occupancy as opposed to multiple occupancy dwellings by a ratio of 2 to 1 over the period studied. The majority of the accidental dwelling fire smoke / toxic fumes inhalation injuries occurred in areas with the highest level of deprivation within the area studied. Alcohol / drug consumption was a contributory factor in roughly 14% of the smoke / toxic fumes inhalation injuries. Smoke / toxic fumes inhalation injuries sustained attempting to fight the fire constituted roughly 12% of the total number of smoke / toxic fumes inhalation injuries over the period studied.

The etiology and mortality of altered level of consciousness in the emergency room: before and after coronavirus disease

Background: Coronavirus disease 2019 (COVID-19) has resulted in social, economic, medical, and psychological changes. New-onset altered level of consciousness (ALC) is a classical presentation in real-world medicine. This study investigated changes in ALC in the emergency room (ER) in the periods before (BC) and after (AC) COVID-19.Methods: This was a retrospective study of patients with ALCs who visited the ER of a tertiary referral center, and their medical records BC and AC were compared. A consortium allocated and analyzed the etiologies of ALC in a case-by-case discussion. The time point for etiological assessment was the time of discharge from the ER.Results: In total, 1,936 patients with ALCs (731 and 1,205 in BC and AC, respectively) were investigated. The most common etiology was systemic infection (25.9%), followed by metabolic causes (20.8%). Systemic infections (22.9% vs. 30.8%, P&lt;0.001) and stroke (14.6% vs. 18.2%, P=0.037) were lower in AC than in BC, respectively, whereas rates of toxicity (15.4% vs. 6.0%, P&lt;0.001) and traumatic brain injury (TBI; 5.9% vs. 0.8%, P&lt;0.001) were higher in AC than in BC. The overall mortality rate of ALC in the ER was 18.5%.Conclusion: This study demonstrated that the major etiologies of ALC in the ER were extra-cranial (58.5%). The mortality of ALC in the ER and the incidence of toxic cause and TBI increased in AC, suggesting a change in medical circumstances after the Pandemic.

Melatonin protects against ketorolac induced gastric mucosal toxic injuries through molecular mechanism associated with the modulation of Arylakylamine N-Acetyltransferase (AANAT) activity

Ketorolac tromethamine (KT), is a widely used non-steroidal anti-inflammatory drug (NSAID) for treating moderate to severe pain. However, the use of KT has been restricted due to its highly toxic attributes that lead to severe gastric ulceration and bleeding. The protective effects of exogenous melatonin (MT) has been reported in conditions associated with gastro-intestinal disorders. This study aims at exploring the role of gastric endogenous MT level and it's metabolizing enzyme AANAT, at the onset of ketorolac mediated toxicities in the gastric mucosa. Gastric mucosal damage was induced in experimental rats by oral administration of graded doses of KT, where 50 mg/kg b.w. of KT was observed to incur maximum gastric lesions. However, gastric damages were found to be protected in rats, pre-treated with 60 mg/kg b.w. of MT. Post-sacrifice, mean ulcer index, oxidative status, total melatonin levels and enzyme activities associated with MT biosynthesis and catabolism were estimated. The results reveal that KT decreases AANAT activity with a concomitant decline in endogenous MT level which cumulatively aggravates gastric toxicity. Moreover, exogenous MT administration has been found to be protective in ameliorating this ulcerogenic process in rats, challenged with KT. Biochemical and histo-pathological observations revealed the reduction in oxidative stress level and replenishment of depleted gastric MT levels in MT pre-treated animals, which might be the causative factors in conferring protection to the gastric tissues and residing mitochondria. The results revealed a correlation between depleted gastric MT level and ulcer formation, which unveiled a novel ulcerogenic mechanism. This may bring forth future therapeutic relevance for treating patients suffering from KT mediated acute gastric toxicities.

#3971 INJURY INDUCED RENAL FIBROSIS PROMOTES CYSTOGENESIS AND CYST GROWTH IN ADULT MICE WITH AUTOSOMAL DOMINANT POLYCYSTIC KIDNEY DISEASE

Abstract Background and Aims Acute kidney injury (AKI) is not a merely transient event, which can lead to a subsequent progression to chronic kidney disease (CKD). Autosomal dominant polycystic kidney disease (ADPKD) is the most frequent inherited kidney disease caused by mutations in PKD1 or PKD2 gene. Previous studies showed that acute kidney injuries promote cystogenesis in adult mice with ADPKD. Fibrosis is a hallmark of AKI to CKD (AKI-CKD) transition, and positively correlated with renal cyst growth in humans. All these data imply a role of AKI-CKD transition in ADPKD.In this study, we aimed to determine the role of AKI-CKD transition in the ADPKD. Method We established AKI-CKD transition model by toxic or surgical injuries, and Pkd gene inactivation in adult mice was performed in different time point of AKI-CKD transition. The progression of renal fibrosis was shown by Western blotting analysis in toxic or surgical injury induced AKI-CKD transition model. Results We showed that renal injury before or after Pkd gene inactivation can both induce renal cyst formation in adult Pkd1 or Pkd2 mice, and the extent of cyst formation was correlated with the initial fibrosis level of three hits (injury and gene inactivation). Inactivation of Pkd1 gene at fibrosis recovery stage in surgery induced less cyst formation in adult Pkd1 mice. Enhanced renal fibrosis by repeated toxic injuries before gene inactivation accelerated renal cyst growth in Pkd1 mice. We further showed that the speed of cyst growth at the early stage in adult Pkd1 mice was decided by the baseline of renal fibrosis. Finally, we showed that conditional knockout of Ezh2 gene attenuated renal fibrosis and cyst growth in adult Pkd1 mice with established renal fibrosis. Conclusion Fibrotic responses in AKI-CKD transition is a driving force for renal cyst formation and growth in adult kidneys and inhibition of renal fibrosis through targeting EZH2 might be new therapeutic strategy for adult ADPKD.

#4369 MYELOID CELL-DERIVED HMGB1 AGGRAVATES THE PROGRESSION OF CHRONIC KIDNEY DISEASE INDUCED BY CHRONIC OXALATE EXPOSURE

Abstract Background and Aims HMGB1, long form High Mobility Group Box 1, a highly conserved DNA chaperone stabilizes chromatin in the nucleus, regulates gene transcription, and supports DNA repair. In the cytosol, it regulates mitochondrial function, autophagy, and apoptosis. In addition, when secreted or passively released into the extracellular space during cell necrosis, it acts as danger signal (DAMP). As such HMGB1 amplifies necroinflammation in ischemic and toxic acute kidney injury. In contrast, little is known about the contribution of HMGB1 release from infiltrating immune cells in this context. We hypothesized that HMGB1 from resident and infiltrating myeloid cells would contribute to the progression of CKD by driving tubular atrophy and tissue remodeling. Method We generated myeloid deletion of HMGB1 by crossing Hmgb1 floxed mice (Hmgb1f/f) with endogenous Lyz2 promoter (Lyz2Cre/+) to investigate the role of myeloid cell HMGB1 in a model of chronic kidney disease induced by sodium oxalate-rich diet (50 μmol/g), i.e., calcium oxalate nephropathy. The Lyz2Cre/+ mice were on a C57BL/6J background and crossed with mice homozygous for the floxed Hmgb1 gene to generate mice with a constitutive deletion of the Hmgb1 gene in all myeloid cells. We used littermates of 6- or 8-week-old males for all experiments. Mice with no Lyz2Cre/+ but the homozygous of floxed Hmgb1 gene served as wildtype controls (Lyz2Cre/−Hmgb1f/f WT) and mice with Lyz2Cre/+ and the homozygous of floxed Hmgb1 gene were used as Lyz2Cre/+Hmgb1f/f KO. The primary endpoint for the comparison between Lyz2Cre/+Hmgb1f/f KO and WT mice was glomerular filtration rate at 21 days of oxalate feeding, assessed by FITC-sinistrin clearance. Plasma samples were collected only on day 21 before sacrifice by cervical dislocation. Urine samples were collected as well as GFR was measured from all experimental groups on day 0, day7, day14 and before sacrifice by cervical dislocation on day 21. Kidneys were harvested after sacrifice. The kidney was divided into two equal parts. One part was kept in RNA later solution at −80°C for RNA isolation and the second part was kept in 4% formalin to be embedded in paraffin for histology analysis. Results KO animals did not reveal any abnormalities within an observation phase of 6 months. Healthy mice showed normal kidney function parameters in urine and blood. Observation is continued and will be supplemented by histopathological analyzes at the age of 12 months. Primary endpoint: Deletion of HMGB1 from myeloid cells attenuated the decline of GFR as compared to WT control mice. Secondary endpoints: The difference in GFR was consistent with the respective levels of serum creatinine at day 21 upon oxalate-rich diet compared with controls. The ablation of HMGB1 in myeloid cells was associated with less tubulointerstitial fibrosis at day 21 after oxalate-rich diet Conclusion Our data suggest that HMGB1 release from myeloid cells promotes crystal-induced chronic kidney injury, which is consistent with the role of HMGB1 as an extracellular DAMP and persistent inflammation in CKD. However, this effect may also relate to the intracellular role of HMGB1 as a transcriptional regulator.

Toxic and infectious lung injury differential diagnosis specifics in oncohematological patients

Background. Assessment of lung injury in oncohematological patients is a relevant problem, since the spectrum of pathological changes is wide and includes pulmonary infections, tumor cell infiltration, cardiogenic and non-cardiogenic pulmonary edema, bronchiolitis obliterans, interstitial pneumonitis, post-radiation and post-inflammatory pneumofibrosis, pulmonary vasculopathy and pleural effusion. At the moment there are no approved recommendations with criteria of differential diagnosis for these conditions, in particular, with differences between the most common therapy complication represented by pulmonary infections and poorly explored drug-induced toxic lesions.Aim. Identification of criteria for pneumotoxicity, allowing for differential diagnosis with pulmonary infections developing during chemotherapy, according to data routinely obtained in real clinical practice.Materials and methods. The study group included 38 patients with cytotoxic and autoimmune lung injury caused by specific therapy (group 1); the comparison group included 38 patients with infectious lesions receiving the same antitumor drugs (group 2). The data of the anamnesis, clinical course, instrumental studies and standard laboratory tests was studied retrospectively. For statistical analysis, the Mann–Whitney, χ2, Kruskal–Wallis tests were used. ROC analysis was performed to assess the sensitivity and specificity of various factors in relation to toxic damage.Results. Patients with lymphomas predominated in group of toxic lung injury (63 %). In patients who underwent allogeneic hematopoietic stem cells transplantation, toxic complications developed in the period from 35 to 1289 days, infectious – from 4 to 43 days. Statistically significant differences were obtained in the presence of a concomitant state of an altered immune response: 32 % of patients in the toxic lesion group versus 5 % in the infectious group had a history of allergy, and, in contrast to the infectious lesion group, in the toxic lesion group autoimmune diseases were detected. The main symptom in patients of the first group was shortness of breath, which was observed in 68 % of cases, of the second – an increased body temperature, observed in 92 % of cases; cough was also a common symptom – in 19 % and 13 % of patients respectively. In 58 % of patients of the second group, concomitant mucositis was detected, while in the first group this complication did not occur in any of them. The most common radiological pattern (71 % of cases in each group) was ground-glass opacities, in patients of the second group often combined with infiltrative changes and thickening of the bronchial walls (in 53 and 42 % of cases respectively). Among laboratory results, the largest differences between groups were observed in the leukocyte levels (with an average level of 2.5 . 109 / L in the infectious group versus 6 . 109 / L in the toxic group), eosinophils (with an average of 3.6 % in the toxic group versus 1.75 % in the infectious group), C-reactive protein (with an average level of 146.7 mg / L in the infectious group versus 52.4 mg / L in the toxic group), and creatinine (with an average of 0.085 mmol / L in the toxic group versus 0.071 mmol / L in the infectious group).Conclusion. The data obtained in this research indicates the value of taking an anamnesis and the importance of performing additional studies in patients with suspected drug-induced lung injury, as well as identifies risk groups. Based on the revealed differences, a scale for the differential diagnosis of drug-induced toxic and infectious lung damage, which includes the results of publicly available research methods, with high sensitivity and specificity, was proposed. Further research for more specific, but, at the same time, universal for various drugs, criteria for toxic lung damage is relevant.

Emergencies in Oncological Practice

Oncological emergencies are defined as an acute lifethreatening event in a patient with a tumor occurring as part of their complex treatment regimen or secondarily to their underlying malignancy. These events can occur at any time from the initial diagnosis of their cancer to endstage disease; and can be encountered in any clinical setting, ranging from primary care physician and emergency department visits to a variety of subspecialty environments. Oncologic emergencies are clinical situations that can lead to death in a short time (24-48 hours) if not quickly faced. In the clinical practice of the medical oncologist, such situations do not infrequently occur. The onset of oncologic emergencies may depend on the presence of cancer itself, the therapies carried out to counteract cancer, or the patient’s predisposition to develop such events. It is essential to recognize the aforementioned situations early in order to treat them promptly, thus avoiding serious consequences. BJM Vol. 34 No. 2(1) Suppliment 2023 Scientific Presentation 186 Therefore, it is critically important that all physicians have a working knowledge of the potential oncological emergencies that may present in their practice and how to provide the most effective care without delay. Nervous system emergencies include spinal cord compression, raised ICP, leptomeningeal disease, seizures and altered mental status whereas superior venacaval syndrome, hyperviscocity syndrome, hyperleukocytosis, venous thromboembolism, Hemorrhage and DIC are the vascular and haematologic emergencies. Several classic metabolic oncologic emergencies include syndrome of inappropriate antidiuretic hormone secretion, tumor lysis syndrome and hypercalcemia of malignancy. Among the pulmonary problems airway obstruction, massive haemoptysis, toxic lung injuries, pneumonitis and pulmonary fibrosis can be caused by cancer and cancer treatment. Urologic emergencies such as hemorrhagic cystitis and obstructive uropathy are also seen. Gastrointestinal bleeding in patients with cancer and typlitis in patients with neutropenic fever are potentially serius complications also. Immune check point inhibitors may cause irAEs in practically any organ system ranging from SJS, TEN, thyroiditis, hypophysitis, adrenilitis, diabetic ketoacidosis to life threatening pneumonitis and myocarditis which may be associated with poor overall survival. Oncologic emergencies can threaten the well-being of almost any patient with a malignancy. Although some of these conditions are related to cancer therapy, they are by no means confined to the period of initial diagnosis and active treatment. In the setting of recurrent malignancy, these events can occur years after the surveillance of a cancer patient has been appropriately transferred from a medical oncologist to a primary care provider. As such, awareness of a patient’s cancer history and its possible complications forms an important part of any clinician’s knowledge base. Prompt identification of and intervention in these emergencies can prolong survival and improve quality of life, even in the setting of terminal illness. Bangladesh J Medicine 2023; Vol. 34, No. 2(1) Supplement: 186-187

Label-Free Imaging Techniques to Evaluate Metabolic Changes Caused by Toxic Liver Injury in PCLS.

Abuse with hepatotoxic agents is a major cause of acute liver failure. The search for new criteria indicating the acute or chronic pathological processes is still a challenging issue that requires the selection of effective tools and research models. Multiphoton microscopy with second harmonic generation (SHG) and fluorescence lifetime imaging microscopy (FLIM) are modern label-free methods of optical biomedical imaging for assessing the metabolic state of hepatocytes, therefore reflecting the functional state of the liver tissue. The aim of this work was to identify characteristic changes in the metabolic state of hepatocytes in precision-cut liver slices (PCLSs) under toxic damage by some of the most common toxins: ethanol, carbon tetrachloride (CCl4) and acetaminophen (APAP), commonly known as paracetamol. We have determined characteristic optical criteria for toxic liver damage, and these turn out to be specific for each toxic agent, reflecting the underlying pathological mechanisms of toxicity. The results obtained are consistent with standard methods of molecular and morphological analysis. Thus, our approach, based on optical biomedical imaging, is effective for intravital monitoring of the state of liver tissue in the case of toxic damage or even in cases of acute liver injury.

Cognitive and Cellular Effects of Combined Organophosphate Toxicity and Mild Traumatic Brain Injury.

Traumatic brain injury (TBI) is considered the most common neurological disorder among people under the age of 50. In modern combat zones, a combination of TBI and organophosphates (OP) can cause both fatal and long-term effects on the brain. We utilized a mouse closed-head TBI model induced by a weight drop device, along with OP exposure to paraoxon. Spatial and visual memory as well as neuron loss and reactive astrocytosis were measured 30 days after exposure to mild TBI (mTBI) and/or paraoxon. Molecular and cellular changes were assessed in the temporal cortex and hippocampus. Cognitive and behavioral deficits were most pronounced in animals that received a combination of paraoxon exposure and mTBI, suggesting an additive effect of the insults. Neuron survival was reduced in proximity to the injury site after exposure to paraoxon with or without mTBI, whereas in the dentate gyrus hilus, cell survival was only reduced in mice exposed to paraoxon prior to sustaining a mTBI. Neuroinflammation was increased in the dentate gyrus in all groups exposed to mTBI and/or to paraoxon. Astrocyte morphology was significantly changed in mice exposed to paraoxon prior to sustaining an mTBI. These results provide further support for assumptions concerning the effects of OP exposure following the Gulf War. This study reveals additional insights into the potentially additive effects of OP exposure and mTBI, which may result in more severe brain damage on the modern battlefield.

IRG1 prevents excessive inflammatory responses and cardiac dysfunction after myocardial injury

Acute myocardial infarction (MI) and chemotherapeutic drug administration can induce myocardial damage and cardiomyocyte cell death, and trigger the release of damage-associated molecular patterns (DAMPs) that initiate the aseptic inflammatory response. The moderate inflammatory response is beneficial for repairing damaged myocardium, while an excessive inflammatory response exacerbates myocardial injury, promotes scar formation, and results in a poor prognosis of cardiac diseases. Immune responsive gene 1 (IRG1) is specifically highly expressed in activated macrophages and mediates the production of tricarboxylic acid (TCA) cycle metabolite itaconate. However, the role of IRG1 in the inflammation and myocardial injury of cardiac stress-related diseases remains unknown. Here, we found that IRG1 knockout mice exhibited increased cardiac tissue inflammation and infarct size, aggravated myocardial fibrosis, and impaired cardiac function after MI and in vivo doxorubicin (Dox) administration. Mechanically, IRG1 deficiency enhanced the production of IL-6 and IL-1β by suppressing the nuclear factor red lineage 2-related factor 2 (NRF2) and activating transcription factor 3 (ATF3) pathway in cardiac macrophages. Importantly, 4-octyl itaconate (4-OI), a cell-permeable derivative of itaconate, reversed the inhibited expression of NRF2 and ATF3 caused by IRG1 deficiency. Moreover, in vivo 4-OI administration inhibited the cardiac inflammation and fibrosis, and prevented adverse ventricle remodeling in IRG1 knockout mice with MI or Dox-induced myocardial injury. Our study uncovers the critical protective role of IRG1 in suppressing inflammation and preventing cardiac dysfunction under ischemic or toxic injury conditions, providing a potential target for the treatment of myocardial injury.

Red blood cell trapping in the kidney vasculature promotes mitochondrial dysfunction independent of ischemia time

Red blood cell (RBC) trapping presents as the expansion of- and obstruction, of the outer-medullary capillaries of the kidney with tightly packed RBCs. RBC trapping has been thought to promote hypoxic tubular injury by extending ischemia time to the outer-medulla. Recent evidence from our laboratory; however, indicates that RBC trapping may promote toxic tubular injury, secondary to the mitochondrial uptake of iron from hemoglobin released from damaged RBCs in nearby congested capillaries. In the current study, we tested the hypothesis that ‘RBC trapping promotes tubular mitochondrial dysfunction independent of ischemia’. We tested our hypothesis using 9–10-week-old male Wistar-Kyoto rats. In an initial study, rats underwent bilateral warm arterial ischemia for 45 minutes with 2 hours of reperfusion before harvesting the kidneys for analysis by transmission electron microscopy (n=3). In a second study, the left kidney from each of n=10 rats underwent either arterial, venous or sham control clamping for 45 minutes. An alternate procedure (either arterial, venous or sham clamping) was performed on the right kidney. To examine the effect of ischemia alone on mitochondrial function, renal arteries were clamped, and the kidney excised without releasing the clamp. To examine the effect of ischemia with RBC trapping on mitochondrial function, the renal vein was clamped. Renal venous clamping mimics the obstruction of the medullary venous vasculature that occurs during reperfusion from arterial clamping, resulting in both kidney ischemia and RBC trapping. In all animals body temperature was maintained using a servo-controlled heating table and heat lamp. At harvest the kidney medulla and cortex separated and mitochondria isolated from each region. Mitochondrial membrane potential (TMRM fluorescence) and hydrogen peroxide production (Amplex red) was then determined from n=6-7 kidneys for each intervention. Data were compared using ANOVA. Consistent with iron uptake into mitochondria, when viewed using transmission electron microscopy, following 2 hours of reperfusion from arterial clamping, tubular mitochondria near RBC congested vessels often appeared darkened. Both ischemia alone and ischemia with RBC trapping, resulted in a significant increase in mitochondrial H 2 0 2 production compared to control (7063±562 and 7519±803 vs 5688±389 A.U per mg of protein, respectively, p&lt;0.001). In contrast, ischemia alone resulted in a lower mitochondrial membrane potential (228±18 A.U) compared to control (253±18), whereas, mitochondrial membrane potential following ischemia with RBC trapping (308±26) was significantly greater than both ischemia alone and control (p&lt;0.001). Our data supports the hypothesis that congestion of the kidney vasculature with RBCs directly promotes mitochondrial dysfunction independent of ischemia. This likely occurs secondary to mitochondrial uptake of iron from hemoglobin released from damaged RBCs. NIH NIDDK ISAC 21AU4231, NIH NHLBI P01 HL134604 and an American Heart Association Transformational Project Award 970585 to P.O and NIH NIDDK F31 DK127683 to S.R.M. NIH R01 HL148114 D.I. This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Ataxia Telangiestasia Mutated (ATM) Inhibition Disrupts Nuclear And Mitochondrial Homeostasis In Pediatric Nonalcoholic Fatty Liver Disease (PNAFLD)

BACKGROUND: The pathophysiology of PNAFLD is important to understand in order to improve outcomes. Integration of critical components including metabolic stress, inflammation, organelle function, and DNA damage and repair should be particularly critical for this purpose. To emphasize the contribution of DNA damage response and cell survival mechanisms on PNAFLD outcomes during the period of active liver growth, we focused on the ATM pathway. In toxic liver injury settings, replenishment of ATM overcomes restrictions in liver homeostasis and regeneration. METHODS and RESULTS: We undertook a study of PNAFLD in a cohort of children under 12 years of age to represent the period of active liver growth (N=47). Demographic details were noted including age, gender, ethnic origin and duration of obesity as defined by BMI greater than 85th percentile. B iopsies were examined for NAFLD activity score (NAS) components steatosis, ballooning and inflammation as well as fibrosis. Mitochondrial DNA content was quantitated by qPCR for the mitochondrial gene, TL1, vs the nuclear gene, RNAseP. Ploidy class distributions was evaluated by cytofluorimetric analysis of nuclei stained with Hoechst dye. Cell growth-arrest was examined by immunostaining of p21. ATM expression was quantitated by qPCR for ATM relative to GAPDH. Mitochondrial homeostasis in mice with PNAFLD was evaluated by electron microscopy and Western Blot of fission and fusion proteins. Of the 47 children in the cohort, 74% were male, 26% female; 85% hispanic, 11% white, black or asian, and 4% unspecified. Mean BMI of the cohort was 94±7th percentile. The duration of obesity until time of liver biopsy was 4±2 years. 61% displayed simple steatosis or fatty liver (FL) with NAS less than 5, and 39% had steatohepatitis (NASH), with NAS equal to or greater than 5. Significantly, 50% of children in the cohort had fibrosis at the time of liver biopsy. Disease progression was associated with dyregulated ATM expression as evidenced by less ATM mRNA expression in FL compared to healthy controls (obtained from NIDDK supported biorepository LTPADS from the University of Minnesota) which further declined with NASH. This was associated with greater degree of mitochondrial DNA depletion in NASH vs FL, compared to healthy controls, which is dependent on ATM. Consequences of mitochondrial depletion extended to increased p21 mediated cell growth arrest and pathological polyploidy. In modelling of PNAFLD in mice, exposure of weanling mice to high fat diet plus low dose CCl4 plus ATM kinase antagonism for 4 weeks reproduced hepatic steatosis, inflammation ballooning and fibrogenesis. Moreover, there was disruption of mitochondrial homeostatis including fission and fusion. CONCLUSION: Disruption of ATM is particularly critical for PNAFLD in early liver growth stage by increasing susceptibility to nuclear and mitochondrial injury. These insights should advance translational and clinical studies in this area. 1K08DK125881-01A1 This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Repurposing of sevelamer as a novel antidote against aluminum phosphide poisoning: An in vivo evaluation

Aluminum phosphide (AlP) is widely used for protecting grains from pests. AlP releases toxic phosphine gas (PH3) while exposed to humidity. Poisoning with these tablets is dangerous and can cause death or serious injuries. Up to now, no definite antidote has been introduced for specific treatment of this poisoning. Sevelamer carbonate or sevelamer hydrochloride (Renagel) is a polymeric pharmaceutical prescribed for treating hyperphosphatemia in patients with chronic kidney disease. Sevelamer can bind with phosphate groups and act as an anion exchanger. Herein, sevelamer is repurposed as a potent antidote agent in phosphine gas poisoning. In vivo evaluation was conducted on male Sprague Dawley rats. The evaluation was conducted on three groups of animals: control, AlP-poisoned, and AlP-poisoned treated with sevelamer. Survival percentage, serum biomarkers level of organ injury, and ATP level were recorded. The results indicate a high survival rate in sevelamer-treated animals compared with the AlP-poisoned group (75% vs. 0% respectively, 48 h after poisoning). The analysis of serum markers of organ injury also showed that sevelamer could reduce toxicity and organ injury in poisoned animals. ATP level of separate organs showed that sevelamer treated groups were recovered. The results showed that sevelamer could be a potent antidote for managing aluminum phosphide poisoning. Moreover, a mechanism is suggested for the interaction of sevelamer with phosphine gas.

Bioenergetics: the evolutionary basis of progressive kidney disease.

Chronic kidney disease (CKD) affects >10% of the world population, with increasing prevalence in middle age. The risk for CKD is dependent on the number of functioning nephrons through the life cycle, and 50% of nephrons are lost through normal aging, revealing their vulnerability to internal and external stressors. Factors responsible for CKD remain poorly understood, with limited availability of biomarkers or effective therapy to slow progression. This review draws on the disciplines of evolutionary medicine and bioenergetics to account for the heterogeneous nephron injury that characterizes progressive CKD following episodes of acute kidney injury with incomplete recovery. The evolution of symbiosis in eukaryotes led to the efficiencies of oxidative phosphorylation and the rise of metazoa. Adaptations to ancestral environments are the products of natural selection that have shaped the mammalian nephron with its vulnerabilities to ischemic, hypoxic, and toxic injury. Reproductive fitness rather than longevity has served as the driver of evolution, constrained by available energy and its allocation to homeostatic responses through the life cycle. Metabolic plasticity has evolved in parallel with robustness necessary to preserve complex developmental programs, and adaptations that optimize survival through reproductive years can become maladaptive with aging, reflecting antagonistic pleiotropy. Consequently, environmental stresses promote trade-offs and mismatches that result in cell fate decisions that ultimately lead to nephron loss. Elucidation of the bioenergetic adaptations by the nephron to ancestral and contemporary environments may lead to the development of new biomarkers of kidney disease and new therapies to reduce the global burden of progressive CKD.

Acute Toxicity of 6PPD-Quinone to Early Life Stage Juvenile Chinook (Oncorhynchus tshawytscha) and Coho (Oncorhynchus kisutch) Salmon.

The breakdown product of the rubber tire antioxidant N-(1,3-dimethylbutyl)-N'-phenyl-p-phenylenediamine-quinone (6PPD)-6-PPD-quinone has been strongly implicated in toxic injury and death in coho salmon (Oncorhynchus kisutch) in urban waterways. Whereas recent studies have reported a wide range of sensitivity to 6PPD-quinone in several fish species, little is known about the risks to Chinook salmon (Oncorhynchus tshawytscha), the primary prey of endangered Southern Resident killer whales (Orcinus orca) and the subject of much concern. Chinook face numerous conservation threats in Canada and the United States, with many populations assessed as either endangered or threatened. We evaluated the acute toxicity of 6PPD-quinone to newly feeding (~3 weeks post swim-up) juvenile Chinook and coho. Juvenile Chinook and coho were exposed for 24 h under static conditions to five concentrations of 6PPD-quinone. Juvenile coho were 3 orders of magnitude more sensitive to 6PPD-quinone compared with juvenile Chinook, with 24-h median lethal concentration (LC50) estimates of 41.0 and more than 67 307 ng/L, respectively. The coho LC50 was 2.3-fold lower than what was previously reported for 1+-year-old coho (95 ng/L), highlighting the value of evaluating age-related differences in sensitivity to this toxic tire-related chemical. Both fish species exhibited typical 6PPD-quinone symptomology (gasping, increased ventilation, loss of equilibrium, erratic swimming), with fish that were symptomatic generally exhibiting mortality. The LC50 values derived from our study for coho are below concentrations that have been measured in salmon-bearing waterways, suggesting the potential for population-level consequences in urban waters. The higher relative LC50 values for Chinook compared with coho merits further investigation, including for the potential for population-relevant sublethal effects. Environ Toxicol Chem 2023;42:815-822. © 2023 His Majesty the King in Right of Canada and The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. Reproduced with the permission of the Minister of Fisheries and Oceans Canada.

Pathomorphological changes in the organs of laboratory animals with experimental liver cirrhosis

Objective. Develop an experimental model of liver cirrhosis and describe pathomorphological changes in organs and tissues of laboratory animals.Materials and methods. Liver cirrhosis was simulated on 11 sexually mature Wistar male white rats. The control group consisted of 12 healthy laboratory animals. Acute toxic liver injury resulting in cirrhosis was induced by intraperitoneal injection of 50% tetrachloromethane (CCl4) solution on olive oil on the first day of the experiment in the dosage of 0.1 ml CCl4 + 0.4 ml olive oil per 100 g of the animal body weight, on the second day of the experiment 0.3 ml CCl4 + 0.2 ml olive oil per 100 g of the animal body weight. To synergize and potentiate the hepatotoxic effect of CCl4, animals had 10% ethanol solution freely available daily. The duration of the experiment was 65 days. Clinical and laboratory parameters were evaluated, histological evaluation of the preparations was performed. The obtained data were compared with the control group of animals.Results. Reproducibility of the model was 82% (9 animals). The indicator of lymphocyte blabbing index showed the development of liver cirrhosis in animals of the experimental group. Comprehensive morphological confirmation of the developed liver cirrhosis was obtained.Conclusion. The proposed methodology of liver cirrhosis formation correlates with pathomorphological changes in lymphocyte cell wall, changes in liver tissue and internal organs in human liver cirrhosis.

15 Ghost Toxicity and Acute Kidney Injury

15 Ghost Toxicity and Acute Kidney Injury

Appraisal of the Pre-Emptive Effect of Lactoferrin Against Chromium-Induced Testicular Toxicity in Male Rats

Lactoferrin (LCF), a potent naturally occurring antioxidant, is a crucial component in preventing potassium dichromate (PDC) toxicity. The goal of the current work was to study the potential efficacy of LCF in preventing PDC(CrVI)-induced testicular toxicity and oxidative injury in rats. Six groups of male rats of Wistar stain were randomly categorized into: group 1, which served as the control; group 2 and 3 received LCF (200 and 300 mg/kg orally, respectively); group 4 received PDC (2 mg/kg i.p.); group 5 and 6 pretreated with LCF, followed by PDC as in group 4 with 90 min apart for 28 days. PDC-intoxicated rats showed a significantly altered spermogram with abnormal sperm morphology. PDC significantly upregulated serum FSH and downregulated testosterone levels. Additionally, PDC decreased the levels of testicular key antioxidant biomarkers (catalase (CAT), superoxide dismutase (SOD), and glutathione (GSH)) with elevated lipid peroxidation marker (TBARS) and testicular chromium content. Moreover, it upregulated testicular proinflammatory cytokines, IL-1, IL-6, IL-10, and TNF-α, induced histopathological changes in testes with significant immunohistochemical expression of FasL and moderate expression of Nrf2. Pretreatment with LCF significantly mitigated PDC-induced testicular toxicity by enhancing spermogram, improving hormonal levels, restoring testicular oxidant/antioxidant balance, and decreasing testicular IL-1, IL6, IL-10, and TNFα levels, and amending both FasL and Nrf2 immunohistochemical-expression. Additionally, LCF improved testicular histopathological picture and spermatogenesis. Our results highlight the importance of LCF as a superior protective modulator of PDC-induced testicular injury.

Role of Mitochondria and Mitochondrial Transplantation in Drug-induced Toxic Organ Injury

Mitochondria, which are one of the main organelles of the cell, have vital importance for the body. Mitochondrial mechanisms, which have critical roles in many physiological processes, are active in drug-induced toxic tissue damage as well as in diseases related to mitochondrial dysfunction. Mitochondrial dysfunction is a major mechanism by which various drugs can cause adverse effects in various tissues such as the liver, kidney and heart. Inhibiting respiratory complexes of the electron chain; disrupting cell bioenergetic mechanisms; inducing mitochondrial oxidative stress; inhibiting DNA replication, transcription, or translation; and reduction of protein synthesis are the most common ways drugs harm mitochondria. Mitochondrial transplantation has emerged as a promising area that has been studied more frequently in recent years. The importance of mitochondrial transplantation in a variety of mitochondrial dysfunction-related diseases such as cardiovascular diseases, neurodegenerative diseases, and ischemia has been emphasized. The purpose of this review article is to present current information on the role of mitochondria in toxic drug damage and the possible effects of mitochondrial transplantation on toxic damage.