Repeated exposure to eucalyptus wood smoke alters pulmonary gene and metabolic profiles in male Long-Evans rats.

Exposure to wildfire smoke is associated with both acute and chronic cardiopulmonary illnesses, which are of special concern for wildland firefighters who experience repeated exposure to wood smoke. It is necessary to better understand the underlying pathophysiology by which wood smoke exposure increases pulmonary disease burdens in this population. We hypothesize that wood smoke exposure produces pulmonary dysfunction, lung inflammation, and gene expression profiles associated with future pulmonary complications. Male Long-Evans rats were intermittently exposed to smoldering eucalyptus wood smoke at two concentrations, low (11.0 ± 1.89 mg/m3) and high (23.7 ± 0.077 mg/m3), over a 2-week period. Whole body plethysmography was measured intermittently throughout. Lung tissue and lavage fluid were collected 24 hours after the final exposure for transcriptomics and metabolomics. Increasing smoke exposure upregulated neutrophils and select cytokines in the bronchoalveolar lavage fluid. In total, 3,446 genes were differentially expressed in the lungs of rats in the high smoke exposure and only one gene in the low smoke exposure (Cd151). Genes altered in the high smoke group reflected changes to the Eukaryotic Initiation Factor 2 (EIF2) stress and oxidative stress responses, which mirrored metabolomics analyses. xMWAS-integrated analysis revealed that smoke exposure significantly altered pathways associated with oxidative stress, lung morphogenesis, and tumor proliferation pathways. These results indicate that intermittent, 2-week exposure to eucalyptus wood smoke leads to transcriptomic and metabolic changes in the lung that may predict future lung disease development. Collectively, these findings provide insight into cellular signaling pathways that may contribute to the chronic pulmonary conditions observed in wildland firefighters.

Acute hepatic rupture causing hemoperitoneum in a dog with anaphylaxis.

A 2-year-old spayed female Siberian Husky was presented with a history of acute onset lethargy, collapse, haematochezia and vomiting. The patient was severely tachycardic and hypotensive. Point-of-care ultrasound revealed gallbladder wall thickening and peritoneal effusion consistent with haemorrhage on subsequent abdominocentesis. Despite attempted medical stabilization over the course of several hours, including blood products and multiple autotransfusions, the patient progressed to cardiopulmonary arrest. The dog was successfully resuscitated but was subsequently euthanized. Necropsy revealed a severe, acute hemoperitoneum secondary to rupture of the left lateral liver lobe. A tear in the hepatic capsule was identified along with a large hematoma. A single adult nematode, consistent with Dirofilaria immitis, was found in a pulmonary vessel in the right caudal lung lobe. The remaining necropsy findings were supportive of the clinical diagnosis of anaphylaxis. This report details a case, with necropsy findings, supporting a diagnosis of anaphylaxis and severe, refractory hemoperitoneum resulting from hepatic rupture. Acute hepatic rupture should be considered in cases of anaphylaxis-related hemoperitoneum.

Evaluation of hemostatic derangements associated with canine anaphylaxis and the relationship to syndrome severity.

OBJECTIVE: To describe hemostatic derangements associated with canine anaphylaxis and to assess for association with syndrome severity. DESIGN: Prospective observational study. SETTING: University teaching hospital. ANIMALS: Twenty-seven client-owned dogs, recruited from November 2018 to January 2022, diagnosed with anaphylaxis of varying severity were included. Study inclusion required presentation <6 hours after initiation of clinical signs, no medications or history of illness within the prior 2 weeks, lack of comorbidities expected to affect hemostasis, and lack of a disease state that could alternatively explain the clinical presentation. INTERVENTIONS: Blood samples were collected within the first hour of presentation for CBC, serum biochemistry, prothrombin time (PT), activated partial thromboplastin time (aPTT), and viscoelastic coagulation testing for use with a cartridge-based point-of-care device. MEASUREMENTS AND MAIN RESULTS: Clotting time and clot formation time were prolonged, alpha angle and maximum clot firmness were decreased, PT and aPTT were prolonged, and platelet counts were lower in severe cases compared to mild and moderate cases. There were no differences for any parameter between mild and moderate cases. The presence or absence of abdominal effusion was not associated with hemostatic status. CONCLUSIONS: Global hemostatic derangements consistent with hypocoagulability are a prominent feature of severe anaphylaxis in dogs and should be considered for routine evaluation.

Pregnancy-related hemodynamic biomarkers in relation to trimester-specific maternal per - and polyfluoroalkyl substances exposures and adverse birth outcomes.

The fate of environmental chemicals in maternal and fetal tissues might be affected by pregnancy-related hemodynamic changes that occur across gestation. Specifically, hemodilution and renal function are hypothesized to confound associations between per- and polyfluoroalkyl substances (PFAS) exposure measures in late pregnancy with gestational length and fetal growth. We sought to analyze two pregnancy-related hemodynamic biomarkers, creatinine and estimated glomerular filtration rate (eGFR), as confounders of the trimester-specific relationships between maternal serum PFAS concentrations and adverse birth outcomes. Participants were enrolled in the Atlanta African American Maternal-Child Cohort between 2014 and 2020. Biospecimens were collected at up to two timepoints, which were categorized into the 1st trimester (N = 278; 11 mean weeks gestation), 2nd trimester (N = 162; 24 mean weeks gestation), and 3rd trimester (N = 110; 29 mean weeks gestation). We quantified six PFAS in serum, creatinine in serum and urine, and eGFR using the Cockroft-Gault equation. Multivariable regression models estimated the associations between single PFAS and their sum with gestational age at delivery (weeks), preterm birth (PTB, <37 gestational weeks), birthweight z-scores, and small for gestational age (SGA). Primary models were adjusted for sociodemographics. We additionally adjusted for serum creatinine, urinary creatinine, or eGFR in the confounding assessments. An interquartile range increase in perfluorooctanoic acid (PFOA) produced a non-significant reduction in birthweight z-score during the 1st and 2nd trimesters (ß = -0.01 g [95% CI = -0.14, 0.12] and ß = -0.07 g [95% CI = -0.19, 0.06], respectively) whereas the relationship was significant and positive during the 3rd trimester (ß = 0.15 g; 95% CI = 0.01, 0.29). Trimester-specific effects were similar for the other PFAS and adverse birth outcomes, which persisted after adjusting for creatinine or eGFR. The relationships between prenatal PFAS exposure and adverse birth outcomes were not strongly confounded by renal function or hemodilution. However, 3rd trimester samples consistently exhibited different effects than those collected during the 1st and 2nd trimesters.

Metallomics in pulmonary arterial hypertension patients.

Pulmonary arterial hypertension (PAH) prevalence is increasing worldwide, and the prognosis is poor with 5-year survival < 50% in high risk patients. The relationship between metal exposure/essential metal dyshomeostasis and PAH/right ventricular dysfunction is less investigated. The aim of this study is to investigate vegetable consumptions and metal levels between PAH patients and controls. This was a prospective, single center pilot study. Questionnaires were completed by all study subjects (20 PAH patients and 10 healthy controls) on smoking, metal exposure risks, metal supplements, and vegetable consumptions. Blood and urine samples were collected to measure 25 metal levels in blood, plasma, and urine using an X Series II quadrupole inductively coupled plasma mass spectrometry. Statistical analysis was conducted using SAS 9.5 and results with p value < 0.05 were considered significant. Vegetables consumptions (broccoli risk ratio [RR] = 0.4, CI = (0.2, 0.9)], cabbage [RR = 0.2, CI = (0.1, 0.8)], and brussel sprouts [RR = 0.2, CI = (0.1, 0.5)]) are associated with less risks of PAH. In the plasma samples, silver (p < 0.001), and copper (p = 0.002) levels were significantly higher in PAH patients. There was significant positive correlation between cardiac output and cardiac index with plasma levels of silver (r = 0.665, p = 0.001 and r = 0.678 p = 0.001), respectively. There was significant correlation between mixed venous saturation, 6-min walk distance, and last BNP with plasma levels of chromium (r = -0.520, p = 0.022; r = -0.55, p = 0.014; r = 0.463, p = 0.039), respectively. In conclusion, there are significant differences between PAH and control groups in terms of vegetable consumptions and metal concentrations. Silver and chromium levels are correlated with clinical indicators of PAH severities.

Effects of time delay and blood storage methods on analysis of canine venous blood samples with an Element point-of-care analyzer.

BACKGROUND: Manufacturers of point-of-care (POC) analyzers recommend immediate processing and anaerobic collection of blood samples. However, it is not uncommon for clinical scenarios to result in delayed sample processing or room air exposure that could impact the test results. OBJECTIVE: To investigate the effect of time delay and sample storage method on key POC analytes in canine venous blood samples processed with an Element POC analyzer. METHODS: Blood gas analysis was performed on venous blood samples at times 0 (T0), 15, 30, and 60 minutes after sampling using three different storage methods: preheparinized plastic syringes and two different lithium heparin tubes. To determine clinical relevance, results were compared with allowable total error of the respective parameter. Significance was set at P < 0.05. RESULTS: Significant differences between the three storage methods at baseline were found for partial pressure of carbon dioxide (PCO2 ), partial pressure of oxygen (PO2 ), base excess, and total hemoglobin. No significant differences up to T60 were found within collection methods for actual bicarbonate (HCO3 - ), base excess, sodium, potassium, chloride, ionized calcium (iCa), glucose, and BUN. Significant differences within collection methods were found after T0 for creatinine, after 15 minutes for lactate, and after 30 minutes for pH and hematocrit. No significant differences were found for PO2 in samples stored in preheparinized plastic syringes at any time point. CONCLUSIONS: These results suggest that HCO3 - , sodium, potassium, chloride, iCa, glucose, and BUN are comparable within the three storage methods for up to 60 minutes after sampling without resulting in clinically relevant changes.

Obesity reprograms the pulmonary polyunsaturated fatty acid-derived lipidome, transcriptome, and gene-oxylipin networks.

Obesity exacerbates inflammation upon lung injury; however, the mechanisms by which obesity primes pulmonary dysregulation prior to external injury are not well studied. Herein, we tested the hypothesis that obesity dysregulates pulmonary PUFA metabolism that is central to inflammation initiation and resolution. We first show that a high-fat diet (HFD) administered to C57BL/6J mice increased the relative abundance of pulmonary PUFA-containing triglycerides and the concentration of PUFA-derived oxylipins (particularly prostaglandins and hydroxyeicosatetraenoic acids), independent of an increase in total pulmonary PUFAs, prior to onset of pulmonary inflammation. Experiments with a genetic model of obesity (ob/ob) generally recapitulated the effects of the HFD on the pulmonary oxylipin signature. Subsequent pulmonary next-generation RNA sequencing identified complex and unique transcriptional regulation with the HFD. We found the HFD increased pathways related to glycerophospholipid metabolism and immunity, including a unique elevation in B cell differentiation and signaling. Furthermore, we conducted computational integration of lipidomic with transcriptomic data. These analyses identified novel HFD-driven networks between glycerophospholipid metabolism and B cell receptor signaling with specific PUFA-derived pulmonary oxylipins. Finally, we confirmed the hypothesis by demonstrating that the concentration of pulmonary oxylipins, in addition to inflammatory markers, were generally increased in mice consuming a HFD upon ozone-induced acute lung injury. Collectively, these data show that a HFD dysregulates pulmonary PUFA metabolism prior to external lung injury, which may be a mechanism by which obesity primes the lungs to respond poorly to infectious and/or inflammatory challenges.

Use of Citrated Whole Blood for Point-of-Care Viscoelastic Coagulation Testing in Dogs.

Background: A new, portable bedside coagulation monitor (VCM Vet) has provided a user-friendly, cartridge-based method to perform viscoelastic testing. However, the use of native whole blood limits the time to analyze the sample to minutes. The objective of this study is to assess whether citrated whole blood can be utilized with the cartridge-based system and whether the results are comparable to those of native whole blood. A secondary objective is to assess the viability of citrated whole blood results after up to 4 hours of resting. Methods: The study population consisted of 10 healthy mixed breed dogs. Whole blood samples were collected via jugular venipuncture. Blood was immediately transferred to the VCM test cartridge for native whole blood control group analysis per the manufacturer's instructions, and the remainder was used to fill two 3.2% sodium citrate vacutainer tubes. Test group analysis was performed on samples from each tube concurrently after a rest period of 30 min (baseline), 2 h, and 4 h. Citrated whole blood samples were recalcified for analysis immediately prior to introduction into the test cartridge. Data was recorded for all reported parameters. Results from the citrate groups were compared to the control group and to the citrated baseline to assess for differences. Overall results were compared using mixed ANOVA models. Where found, specific differences were evaluated using Tukey's test. Within-sample variation was investigated and reported as median (range). A p < 0.05 was considered significant. Results: Samples were obtained for a total of 10 control runs and 20 citrated whole blood runs. Comparison of controls to the citrated test groups revealed significant differences in CT (p < 0.001) and MCF (p < 0.002). There were no significant differences between test groups compared to citrated baselines for any parameter. Selected median coefficients of variation were 6.8% (0-68.8%) for CT, 2.4% (0-19.46%) for alpha angle, 3.2% (0-27.4%) for MCF, and 0% (0-16.3%) for 45-min LY45. Conclusion: Citrated whole blood samples can be used with the VCM Vet device; however, new reference intervals for use with citrated whole blood will be required. Results using citrated whole blood samples are not significantly different from baseline after up to 4 h of resting.

Plasma Metabolic Phenotypes of HPV-Associated versus Smoking-Associated Head and Neck Cancer and Patient Survival.

BACKGROUND: Metabolic differences between human papillomavirus (HPV)-associated head and neck squamous cell carcinoma (HNSCC) and smoking-associated HNSCC may partially explain differences in prognosis. The former relies on mitochondrial oxidative phosphorylation (OXPHOS) while the latter relies on glycolysis. These differences have not been studied in blood. METHODS: We extracted metabolites using untargeted liquid chromatography high-resolution mass spectrometry from pretreatment plasma in a cohort of 55 HPV-associated and 82 smoking-associated HNSCC subjects. Metabolic pathway enrichment analysis of differentially expressed metabolites produced pathway-based signatures. Significant pathways (P < 0.05) were reduced via principal component analysis and assessed with overall survival via Cox models. We classified each subject as glycolytic or OXPHOS phenotype and assessed it with survival. RESULTS: Of 2,410 analyzed metabolites, 191 were differentially expressed. Relative to smoking-associated HNSCC, bile acid biosynthesis (P < 0.0001) and octadecatrienoic acid beta-oxidation (P = 0.01), were upregulated in HPV-associated HNSCC, while galactose metabolism (P = 0.001) and vitamin B6 metabolism (P = 0.01) were downregulated; the first two suggest an OXPHOS phenotype while the latter two suggest glycolytic. First principal components of bile acid biosynthesis [HR = 0.52 per SD; 95% confidence interval (CI), 0.38-0.72; P < 0.001] and octadecatrienoic acid beta-oxidation (HR = 0.54 per SD; 95% CI, 0.38-0.78; P < 0.001) were significantly associated with overall survival independent of HPV and smoking. The glycolytic versus OXPHOS phenotype was also independently associated with survival (HR = 3.17; 95% CI, 1.07-9.35; P = 0.04). CONCLUSIONS: Plasma metabolites related to glycolysis and mitochondrial OXPHOS may be biomarkers of HNSCC patient prognosis independent of HPV or smoking. Future investigations should determine whether they predict treatment efficacy. IMPACT: Blood metabolomics may be a useful marker to aid HNSCC patient prognosis.

Multiomics Analysis of Structural Magnetic Resonance Imaging of the Brain and Cerebrospinal Fluid Metabolomics in Cognitively Normal and Impaired Adults.

INTRODUCTION: Integrating brain imaging with large scale omics data may identify novel mechanisms of mild cognitive impairment (MCI) and early Alzheimer's disease (AD). We integrated and analyzed brain magnetic resonance imaging (MRI) with cerebrospinal fluid (CSF) metabolomics to elucidate metabolic mechanisms and create a "metabolic map" of the brain in prodromal AD. METHODS: In 145 subjects (85 cognitively normal controls and 60 with MCI), we derived voxel-wise gray matter volume via whole-brain structural MRI and conducted high-resolution untargeted metabolomics on CSF. Using a data-driven approach consisting of partial least squares discriminant analysis, a multiomics network clustering algorithm, and metabolic pathway analysis, we described dysregulated metabolic pathways in CSF mapped to brain regions associated with MCI in our cohort. RESULTS: The multiomics network algorithm clustered metabolites with contiguous imaging voxels into seven distinct communities corresponding to the following brain regions: hippocampus/parahippocampal gyrus (three distinct clusters), thalamus, posterior thalamus, parietal cortex, and occipital lobe. Metabolic pathway analysis indicated dysregulated metabolic activity in the urea cycle, and many amino acids (arginine, histidine, lysine, glycine, tryptophan, methionine, valine, glutamate, beta-alanine, and purine) was significantly associated with those regions (P < 0.05). CONCLUSION: By integrating CSF metabolomics data with structural MRI data, we linked specific AD-susceptible brain regions to disrupted metabolic pathways involving nitrogen excretion and amino acid metabolism critical for cognitive function. Our findings and analytical approach may extend drug and biomarker research toward more multiomics approaches.

Plasma acylcarnitine levels increase with healthy aging.

Acylcarnitines transport fatty acids into mitochondria and are essential for ß-oxidation and energy metabolism. Decreased mitochondrial activity results in increased plasma acylcarnitines, and increased acylcarnitines activate proinflammatory signaling and associate with age-related disease. Changes in acylcarnitines associated with healthy aging, however, are not well characterized. In the present study, we examined the associations of plasma acylcarnitines with age (range: 20-90) in 163 healthy, non-diseased individuals from the predictive medicine research cohort (NCT00336570) and tested for gender-specific differences. The results show that long-chain and very long-chain acylcarnitines increased with age, while many odd-chain acylcarnitines decreased with age. Gender-specific differences were observed for several acylcarnitines, e.g., eicosadienoylcarnitine varied with age in males, and hydroxystearoylcarnitine varied in females. Metabolome-wide association study (MWAS) of age-associated acylcarnitines with all untargeted metabolic features showed little overlap between genders. These results show that plasma concentrations of acylcarnitines vary with age and gender in individuals selected for criteria of health. Whether these variations reflect mitochondrial dysfunction with aging, mitochondrial reprogramming in response to chronic environmental exposures, early pre-disease change, or an adaptive response to healthy aging, is unclear. The results highlight a potential utility for untargeted metabolomics research to elucidate gender-specific mechanisms of aging and age-related disease.

Mortality rate and prognostic factors for dogs with severe anaphylaxis: 67 cases (2016-2018).

OBJECTIVE: To determine mortality rates for dogs with severe anaphylaxis and identify potential prognostic factors. ANIMALS: 67 dogs with suspected anaphylaxis graded as severe. PROCEDURES: Dogs were classified on the basis of outcome as survivors and nonsurvivors. Medical records were reviewed, and data were extracted including signalment, examination findings, time to hospital admission from onset of clinical signs, CBC results, serum biochemical analysis results, coagulation testing results, and findings on abdominal ultrasonography. Initial treatment within the first 6 hours after hospital admission was recorded for analysis, specifically including the use of epinephrine, diphenhydramine, corticosteroids, antimicrobials, fresh-frozen plasma, and supplemental dextrose. RESULTS: The overall mortality rate was 14.9% (10/67) for dogs with anaphylaxis graded as severe. Serum phosphorus concentration and prothrombin time (PT) were significantly higher in nonsurvivors, compared with survivors. Nonsurvivors had lower presenting body temperatures than survivors. Serum phosphorus concentration ≥ 12.0 mmol/L, hypoglycemia within 6 hours after hospital admission, high PT value, concurrently high PT and partial thromboplastin time (PTT) values > 50% above the reference range limit, and the need for supplemental dextrose were associated with death. The incidences of coagulopathy and peritoneal effusion were unexpectedly high (85.2% and 65.5% of dogs, respectively) but were not indicative of survival. CONCLUSIONS AND CLINICAL RELEVANCE: Despite the poor presenting clinical condition seen in dogs with severe anaphylaxis, the rate of survival with treatment was fairly high. Coagulopathy and the presence of peritoneal effusion were common findings in dogs with severe anaphylaxis. Serum phosphorus concentration ≥ 12.0 mmol/L, high PT value, concurrent increases of PT and PTT values > 50% above reference range limits, hypoglycemia within 6 hours after hospital admission, and the need for supplemental dextrose were associated with death.

Severe Rhabdomyolysis Associated with Acute Amphetamine Toxicosis in a Dog.

A 3-year-old female spayed rat terrier presented for hyperactivity and repetitive circling to the right of less than one-hour duration. On examination, the patient was dehydrated, hyperactive, and dysphoric. Laboratory tests initially revealed elevations in creatine kinase (CK) and aspartate aminotransferase (AST). Serial chemistries indicated significant progression of CK elevation to a maximum of 181,900 U/L on day 3 along with the development of profuse myoglobinuria. A urine drug screening test was positive for amphetamine metabolites. This patient was treated with sedatives, aggressive fluid diuresis, and antioxidants. The dog recovered uneventfully with no indicators of renal dysfunction based on serial blood chemistries and was discharged five days after presentation. Follow-up blood chemistries taken four days after discharge revealed near normalization of CK and resolution of myoglobinuria. This case report describes a particularly severe case of rhabdomyolysis associated with amphetamine toxicity and its successful treatment.

Sex Hormone Regulation of Proteins Modulating Mitochondrial Metabolism, Dynamics and Inter-Organellar Cross Talk in Cardiovascular Disease.

Cardiovascular disease (CVD) is the leading cause of death in the U.S. and worldwide. Sex-related disparities have been identified in the presentation and incidence rate of CVD. Mitochondrial dysfunction plays a role in both the etiology and pathology of CVD. Recent work has suggested that the sex hormones play a role in regulating mitochondrial dynamics, metabolism, and cross talk with other organelles. Specifically, the female sex hormone, estrogen, has both a direct and an indirect role in regulating mitochondrial biogenesis via PGC-1α, dynamics through Opa1, Mfn1, Mfn2, and Drp1, as well as metabolism and redox signaling through the antioxidant response element. Furthermore, data suggests that testosterone is cardioprotective in males and may regulate mitochondrial biogenesis through PGC-1α and dynamics via Mfn1 and Drp1. These cell-signaling hubs are essential in maintaining mitochondrial integrity and cell viability, ultimately impacting CVD survival. PGC-1α also plays a crucial role in inter-organellar cross talk between the mitochondria and other organelles such as the peroxisome. This inter-organellar signaling is an avenue for ameliorating rampant ROS produced by dysregulated mitochondria and for regulating intrinsic apoptosis by modulating intracellular Ca2+ levels through interactions with the endoplasmic reticulum. There is a need for future research on the regulatory role of the sex hormones, particularly testosterone, and their cardioprotective effects. This review hopes to highlight the regulatory role of sex hormones on mitochondrial signaling and their function in the underlying disparities between men and women in CVD.

Environmental Cadmium Enhances Lung Injury by Respiratory Syncytial Virus Infection.

Cadmium (Cd) is a naturally occurring environmental toxicant that disrupts mitochondrial function at occupational exposure levels. The impacts of Cd exposure at low levels through dietary intake remain largely uncharacterized. Human respiratory syncytial virus (RSV) causes severe morbidity, which can require hospitalization and result in death in young children and elderly populations. The impacts of environmental Cd exposure on the severity of RSV disease are unknown. Herein, we used a mouse model to examine whether Cd pre-exposure at a level of dietary intake potentiates pulmonary inflammation on subsequent infection with RSV. Mice were given Cd or saline in drinking water for 28 days. Subsets of these mice were infected with RSV at 5 days before the end of the study. Cd pre-exposure caused relatively subtle changes in lung; however, it elevated the IL-4 level and altered metabolites associated with fatty acid metabolism. After RSV infection, mice pre-exposed to Cd had elevated lung RSV titer and increased inflammation, as measured by histopathology, immune cell infiltration, cytokines, and chemokines. RSV infection after Cd pre-exposure also caused widespread perturbation in metabolism of glycerophospholipids and amino acids (Trp, Met, and Cys, branched-chain amino acids), as well as carnitine shuttle associated with mitochondrial energy metabolism. The results show that Cd burden by dietary intake potentiates RSV infection and severe disease with associated mitochondrial metabolic disruption.

Low-dose cadmium potentiates lung inflammatory response to 2009 pandemic H1N1 influenza virus in mice.

Cadmium (Cd) is a toxic, pro-inflammatory metal ubiquitous in the diet that accumulates in body organs due to inefficient elimination. Responses to influenza virus infection are variable, particularly severity of pneumonia. We used a murine model of chronic low-dose oral exposure to Cd to test if increased lung tissue Cd worsened inflammation in response to sub-lethal H1N1 infection. The results show that Cd-treated mice had increased lung tissue inflammatory cells, including neutrophils, monocytes, T lymphocytes and dendritic cells, following H1N1 infection. Lung genetic responses to infection (increasing TNF-α, interferon and complement, and decreasing myogenesis) were also exacerbated. To reveal the organization of a network structure, pinpointing molecules critical to Cd-altered lung function, global correlations were made for immune cell counts, leading edge gene transcripts and metabolites. This revealed that Cd increased correlation of myeloid immune cells with pro-inflammatory genes, particularly interferon-γ and metabolites. Together, the results show that Cd burden in mice increased inflammation in response to sub-lethal H1N1 challenge, which was coordinated by genetic and metabolic responses, and could provide new targets for intervention against lethal inflammatory pathology of clinical H1N1 infection.

Low-dose cadmium disrupts mitochondrial citric acid cycle and lipid metabolism in mouse lung.

Cadmium (Cd) causes acute and chronic lung toxicities at occupational exposure levels, yet the impacts of Cd exposure at low levels through dietary intake remain largely uncharacterized. Health concerns arise because humans do not have an effective Cd elimination mechanism, resulting in a long (10- to 35-y) biological half-life. Previous studies showed increased mitochondrial oxidative stress and cell death by Cd yet the details of mitochondrial alterations by low levels of Cd remain unexplored. In the current study, we examined the impacts of Cd burden at a low environmental level on lung metabolome, redox proteome, and inflammation in mice given Cd at low levels by drinking water (0, 0.2, 0.6 and 2.0 mg Cd/L) for 16 weeks. The results showed that mice accumulated lung Cd comparable to non-smoking humans and showed inflammation in lung by histopathology at 2 mg Cd/L. The results of high resolution metabolomics combined with bioinformatics showed that mice treated with 2 mg Cd/L increased levels of lipids in the lung, accompanied by disruption in mitochondrial energy metabolism. In addition, targeted metabolomic analysis showed that these mice had increased accumulation of mitochondrial carnitine and citric acid cycle intermediates. The results of redox proteomics showed that Cd at 2 mg/L stimulated oxidation of isocitrate dehydrogenase, malate dehydrogenase and ATP synthase. Taken together, the results showed impaired mitochondrial function and accumulation of lipids in the lung with a Cd dose response relevant to non-smokers without occupational exposures. These findings suggest that dietary Cd intake could be an important variable contributing to human pulmonary disorders.

Mitochondrial thiol modification by a targeted electrophile inhibits metabolism in breast adenocarcinoma cells by inhibiting enzyme activity and protein levels.

Many cancer cells follow an aberrant metabolic program to maintain energy for rapid cell proliferation. Metabolic reprogramming often involves the upregulation of glutaminolysis to generate reducing equivalents for the electron transport chain and amino acids for protein synthesis. Critical enzymes involved in metabolism possess a reactive thiolate group, which can be modified by certain oxidants. In the current study, we show that modification of mitochondrial protein thiols by a model compound, iodobutyl triphenylphosphonium (IBTP), decreased mitochondrial metabolism and ATP in MDA-MB 231 (MB231) breast adenocarcinoma cells up to 6 days after an initial 24h treatment. Mitochondrial thiol modification also depressed oxygen consumption rates (OCR) in a dose-dependent manner to a greater extent than a non-thiol modifying analog, suggesting that thiol reactivity is an important factor in the inhibition of cancer cell metabolism. In non-tumorigenic MCF-10A cells, IBTP also decreased OCR; however the extracellular acidification rate was significantly increased at all but the highest concentration (10µM) of IBTP indicating that thiol modification can have significantly different effects on bioenergetics in tumorigenic versus non-tumorigenic cells. ATP and other adenonucleotide levels were also decreased by thiol modification up to 6 days post-treatment, indicating a decreased overall energetic state in MB231 cells. Cellular proliferation of MB231 cells was also inhibited up to 6 days post-treatment with little change to cell viability. Targeted metabolomic analyses revealed that thiol modification caused depletion of both Krebs cycle and glutaminolysis intermediates. Further experiments revealed that the activity of the Krebs cycle enzyme, aconitase, was attenuated in response to thiol modification. Additionally, the inhibition of glutaminolysis corresponded to decreased glutaminase C (GAC) protein levels, although other protein levels were unaffected. This study demonstrates for the first time that mitochondrial thiol modification inhibits metabolism via inhibition of both aconitase and GAC in a breast cancer cell model.

Pilot study on the effect of subcutaneous administration of lactated Ringer's solution on biochemistry parameters in healthy euvolemic cats.

Subcutaneous fluid administration is a commonly used therapy in veterinary practice. Its safety and efficacy have been demonstrated in human clinical studies, but have only rarely been discussed in the veterinary literature. This prospective observational study was performed to evaluate changes during a 24 h period in serum biochemistries associated with administration of lactated Ringer's solution subcutaneously to healthy cats. Lactated Ringer's solution was administered subcutaneously once to ten healthy, euvolemic cats at a dose of 22 mL kg-1. Blood biochemistry analytes were sampled at baseline and at serial time points for a total of 24 h. Changes in biochemical analytes at each time point were compared to baseline and evaluated for statistical significance. Serum blood urea nitrogen (BUN) was significantly less than baseline at 4, 6, 12, 18, and 24 h post-infusion. Serum creatinine was significantly less than baseline at 2, 4 and 6 h. Packed cell volume (PCV) was significantly less than baseline at 6, 12, 18, and 24 h. Total plasma proteins were significantly less than baseline at all time points. Serum electrolytes did not change from baseline at any time point. Urine specific gravity was significantly increased from baseline only at 6 h post-Lactated Ringer's solution (LRS) administration. Subcutaneous administration of lactated Ringer's solution appears to result in haemodilution with minimal change to serum electrolyte concentrations in clinically normal, euvolemic cats.

A novel class of mitochondria-targeted soft electrophiles modifies mitochondrial proteins and inhibits mitochondrial metabolism in breast cancer cells through redox mechanisms.

Despite advances in screening and treatment over the past several years, breast cancer remains a leading cause of cancer-related death among women in the United States. A major goal in breast cancer treatment is to develop safe and clinically useful therapeutic agents that will prevent the recurrence of breast cancers after front-line therapeutics have failed. Ideally, these agents would have relatively low toxicity against normal cells, and will specifically inhibit the growth and proliferation of cancer cells. Our group and others have previously demonstrated that breast cancer cells exhibit increased mitochondrial oxygen consumption compared with non-tumorigenic breast epithelial cells. This suggests that it may be possible to deliver redox active compounds to the mitochondria to selectively inhibit cancer cell metabolism. To demonstrate proof-of-principle, a series of mitochondria-targeted soft electrophiles (MTSEs) has been designed which selectively accumulate within the mitochondria of highly energetic breast cancer cells and modify mitochondrial proteins. A prototype MTSE, IBTP, significantly inhibits mitochondrial oxidative phosphorylation, resulting in decreased breast cancer cell proliferation, cell attachment, and migration in vitro. These results suggest MTSEs may represent a novel class of anti-cancer agents that prevent cancer cell growth by modification of specific mitochondrial proteins.