Esophageal Hypervigilance and Visceral Anxiety Contribute to Symptom Severity of Laryngopharyngeal Reflux.

INTRODUCTION: Laryngopharyngeal reflux (LPR) is a clinical conundrum without a diagnostic gold standard. The Esophageal Hypervigilance and Anxiety Scale (EHAS) is a questionnaire designed for cognitive-affective evaluation of visceral sensitivity. We hypothesized that esophageal hypervigilance and symptom-specific anxiety have an etiopathological role in generation of LPR symptoms, especially when gastroesophageal reflux disease (GERD) cannot explain these symptoms. METHODS: Consecutive patients with LPR and/or GERD symptoms lasting >3 months were prospectively enrolled and characterized using the Reflux Symptom Index, GERD questionnaire, and EHAS. Eligible patients with negative endoscopy underwent 24-hour impedance-pH monitoring off acid suppression for phenotyping GERD and assessment of reflux burden, using conventional metrics (acid exposure time and number of reflux episodes) and novel metrics (mean nocturnal baseline impedance and postreflux swallow-induced peristaltic wave index). RESULTS: Of 269 enrolled patients (mean age 47.1 years, 21-65 years, 60.6% female), 90 patients were with concomitant GERD and LPR symptoms, 32 patients were with dominant LPR symptoms, 102 patients were with dominant GERD symptoms, and 45 were controls. Patients with concomitant GERD and LPR symptoms had higher EHAS than those with dominant GERD symptoms and controls ( P ≤ 0.001); patients with dominant LPR symptoms had higher EHAS than controls ( P = 0.007). On Pearson correlation, EHAS positively correlated with the Reflux Symptom Index. DISCUSSION: Esophageal hypervigilance and symptom-specific anxiety may be more important than reflux burden in LPR symptom perception.

Allergenicity reduction of the bio-elicited peanut sprout powder (BPSP) and toxicological acceptance of BPSP-supplemented diets assessed with ICR mice.

The allergenic and toxicological acceptances of the bio-elicited peanut sprout powder (BPSP) have not been assessed. BPSP was generated from peanut kernels germinated at 26-28 °C for 72 h (designated as 72 h-NGS). The 72 h-NGS were subsequently sliced, incubated, dried, defatted and pulverized to generate bio-elicited peanut sprout powder (BPSP). Protein solubility of BPSP increased 2.6-fold compared to 72 h-NGS. SDS-PAGE analysis revealed BPSP production triggered extensive degradation of the high-molecular weight peanut allergic proteins, mainly Ara h 1 and Ara h 3. Western blotting detected with peanut allergic patients' IgE indicated decreased in vitro reactivity. Food safety assessment of BPSP was performed with ICR mice fed with basal (control) and three doses of formulated BPSP-supplemented diets containing 0.11 g (normal), 2.5 g (high) and 25 g (super high) BPSP /kg BW. Animals appeared healthy with steady body weight gain in all groups during the entire 35-day dietary intervention. Hematological and serum biochemical analyses revealed no significant difference among groups. Histopathological examination on the tissue sections of primary organs further supported safety with no pathologies. The in vitro allergic reduction and toxicological safety in the BPSP-supplemented dietary intervention in the ICR mice study, support moving forward with BPSP-involved product development. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-022-05537-7.

Bovine Kobuvirus in Calves with Diarrhea, United States.

We detected bovine kobuvirus (BKV) in calves with diarrhea in the United States. The strain identified is related genetically to BKVs detected in other countries. Histopathologic findings also confirmed viral infection in 2 BKV cases. Our data show BKV is a potential causative agent for diarrhea in calves.

POSTNATAL MORTALITY IN NEONATE RATTLESNAKES ASSOCIATED WITH OPHIDIOMYCES OPHIODIICOLA.

Ophidiomycosis, historically referred to as snake fungal disease (SFD), caused by Ophidiomyces ophiodiicola, is a significant disease of snakes characterized by crusty scales, pustules, subcutaneous nodules, and death. Ophidiomycosis is a proposed threat to sustainability of free-ranging snake populations throughout the United States and Europe, but the clinical progression during periods of reproductive activity (gravid females, neonates) is unknown. In spring 2012, five apparently healthy gravid eastern massasauga (Sistrurus catenatus) rattlesnakes from Clinton County, Illinois, were brought into captivity to give birth and be returned into the population. While in captivity, one adult female and 21 neonates died. Five individuals were subsequently confirmed positive for O. ophiodiicola by using quantitative polymerase chain reaction (qPCR). In 2016, a gravid timber rattlesnake (Crotalus horridus) with ophidiomycosis from Jackson County, Illinois, gave birth in captivity to 13 neonates. Skin swabs were taken from all neonates immediately after birth and confirmed negative for O. ophiodiicola by using qPCR. The neonates remained housed with the positive female for 10 days before all animals were reswabbed and released back into the wild. One neonate was O. ophiodiicola positive at time of release. The initial negative result followed by a positive result several days postpartum suggests that the neonate was infected by the female after direct contact. Both case series represent natural infection of neonates after parturition and highlight the importance of this disease in a demographically important age class.

PHARMACOKINETICS, EFFICACY, AND SAFETY OF VORICONAZOLE AND ITRACONAZOLE IN HEALTHY COTTONMOUTHS (AGKISTRODON PISCIVORUS) AND MASSASAUGA RATTLESNAKES (SISTRURUS CATENATUS) WITH SNAKE FUNGAL DISEASE.

Snake fungal disease (SFD; Ophidiomyces ophiodiicola) is posing a significant threat to several free-ranging populations of pitvipers. Triazole antifungals have been proposed for the treatment of mycoses in reptiles; however, data are lacking about their safety and efficacy in snakes with SFD. Study 1 investigated in vitro susceptibility, and identified that plasma concentrations >250 ng/ml (voriconazole) and >1,000 ng/ml (itraconazole) may be effective in vivo for SFD. In Study 2, the pharmacokinetics after a single subcutaneous voriconazole injection were assessed in apparently healthy free-ranging cottonmouths (Agkistrodon piscivorus). Based on pilot-study results, four snakes were administered a single injection of voriconazole (5 mg/kg). One pilot snake and three full-study snakes died within 12 hr of voriconazole administration. All surviving snakes maintained plasma concentrations >250 ng/ml for 12-24 hr. In Study 3, two Eastern massasaugas (Sistrurus catenatus) and a timber rattlesnake (Crotalus horridus horridus) diagnosed with SFD were treated with voriconazole delivered by subcutaneous osmotic pumps. The timber rattlesnake (12.1-17.5 mg/kg/hr) reached therapeutic concentrations, whereas the massasaugas (1.02-1.6 mg/kg/hr) did not. In Study 4, the pharmacokinetics of a single 10-mg/kg per-cloaca dose of itraconazole (Sporanox®) was evaluated in seven apparently healthy free-ranging cottonmouths. Similarly, the plasma and tissue concentrations did not meet therapeutic concentrations based on in vitro data. The data presented in this report serve as an initial step toward understanding the pharmacokinetics, efficacy, and safety of triazole antifungals in pitviper species with SFD. Further study is needed to determine the appropriate dose and route of administration of triazole antifungals in pitviper species.

Predicting three-month and 12-month post-fitting real-world hearing-aid outcome using pre-fitting acceptable noise level (ANL).

OBJECTIVE: Determine the extent to which pre-fitting acceptable noise level (ANL), with or without other predictors such as hearing-aid experience, can predict real-world hearing-aid outcomes at three and 12 months post-fitting. DESIGN: ANLs were measured before hearing-aid fitting. Post-fitting outcome was assessed using the international outcome inventory for hearing aids (IOI-HA) and a hearing-aid use questionnaire. Models that predicted outcomes (successful vs. unsuccessful) were built using logistic regression and several machine learning algorithms, and were evaluated using the cross-validation technique. STUDY SAMPLE: A total of 132 adults with hearing impairment. RESULTS: The prediction accuracy of the models ranged from 61% to 68% (IOI-HA) and from 55% to 61% (hearing-aid use questionnaire). The models performed more poorly in predicting 12-month than three-month outcomes. The ANL cutoff between successful and unsuccessful users was higher for experienced (∼18 dB) than first-time hearing-aid users (∼10 dB), indicating that most experienced users will be predicted as successful users regardless of their ANLs. CONCLUSIONS: Pre-fitting ANL is more useful in predicting short-term (three months) hearing-aid outcomes for first-time users, as measured by the IOI-HA. The prediction accuracy was lower than the accuracy reported by some previous research that used a cross-sectional design.

SYSTEMIC BLASTOMYCOSIS IN A CAPTIVE RED RUFFED LEMUR (VARECIA RUBRA).

A 5-yr-old, intact male red ruffed lemur ( Varecia rubra ) presented for evaluation as the result of a 1-wk history of lethargy and hyporexia. Physical examination findings included thin body condition, muffled heart sounds, harsh lung sounds, and liquid brown diarrhea. Complete blood count and serum biochemistry showed an inflammatory leukogram, mild hyponatremia, and mild hypochloremia. Orthogonal trunk radiographs revealed a severe alveolar pattern in the right cranial lung lobes with cardiac silhouette effacement. Thoracic ultrasound confirmed a large, hypoechoic mass in the right lung lobes. Fine-needle aspiration of the lung mass and cytology revealed fungal yeast organisms, consistent with Blastomyces dermatitidis. Blastomyces Quantitative EIA Test on urine was positive. Postmortem examination confirmed systemic blastomycosis involving the lung, tracheobronchial lymph nodes, spleen, kidney, liver, cerebrum, and eye. To the authors' knowledge, this is the first report of blastomycosis in a prosimian species.

A neonatal piglet model reveals interactions between nasal microbiota and influenza A virus pathogenesis.

While vaccination and therapeutics for prevention/treatment of influenza are available, new strategies are needed to combat influenza disease in susceptible populations, particularly young children and newborns. Host associated microbiota play an important role in modulating the virulence of numerous pathogens, including the influenza A virus. In this study, we examined microbiome-influenza interactions in a neonatal piglet model system. The nasal microbiome of newborn piglets was longitudinally sampled before and after intranasal infection with recombinant viruses expressing hemagglutinins (HAs) derived from distinct zoonotic H1 subtypes. We found that viruses expressing different parental HAs manifested unique patterns of pathogenicity, and varied impacts on microbial community diversity. Despite these virus specific differences, a consistent microbial signature of viral infection was detected. Our results indicate that influenza A virus infection associates with the restructuring of nasal microbiome and such shifts in microbial diversity may contribute to outcomes of viral infection in neonatal piglets.

Human exposures to Brucella canis from a pregnant dog during an international flight: Public health risks, diagnostic challenges and future considerations.

AIMS: This report documents the exposure of passengers and crew of a commercial international flight to the zoonotic pathogen Brucella canis after an infected dog aborted in the passenger cabin of the aircraft. This case demonstrates the challenges associated with brucellosis screening and the risks that airline personnel, airport employees and travellers face when animals with unrecognized zoonotic infections are transported. METHODS/RESULTS: The public health investigation of this case was conducted by the Centers for Disease Control, the Illinois Department of Health and the Illinois Department of Agriculture, in collaboration with a local veterinary clinic and several academic and federal diagnostic laboratories. It included an extensive diagnostic evaluation of the dam and aborted foetuses to confirm a diagnosis of canine brucellosis. Passengers, airline personnel and staff from the veterinary clinic where the dogs were treated underwent risk assessments, and clinic staff also received detailed guidance regarding infection prevention practices. CONCLUSIONS: Animal shelters and breeding programs are recommended to screen dogs routinely for brucellosis, but it is not unusual for domestic or imported animals to have unknown health histories, including the dog's brucellosis status, at the time of purchase, adoption, or re-homing. Testing recommendations and requirements vary by state, making it challenging for state public health and animal health agencies to monitor and respond appropriately. This case highlights the importance of Brucella spp. screening in sexually intact dogs prior to breeding, purchase, or domestic or international transportation of the dogs. The transportation of pregnant dogs may present a previously unrecognized public health threat in addition to contributing to unnecessary stress and health risks for pregnant animals.

Association of acute phase protein-haptoglobin, and epithelial-mesenchymal transition in buccal cancer: a preliminary report.

BACKGROUND: The aim of this study was to determine the influence of inflammation on acute phase protein and epithelial-mesenchymal transition (EMT) in buccal cancer. METHODS: Western blotting was carried out to investigate the expression of haptoglobin and epithelial-mesenchymal transition in oral cancer cell lines with or without IL-6 stimulation. We studied patients with buccal cancer patients without distant metastasis at diagnosis. Correlation between cellular haptoglobin, EMT, and clinical characteristics of buccal cancer was analyzed to assess the prognostic value of cellular haptoglobin level and EMT. The relationship of haptoglobin, and EMT expression with survival was assessed using Cox proportional hazard models. RESULTS: Western blotting analysis showed that increased haptoglobin protein was associated with overexpression of vimentin. Under IL-6 stimulation, overexpression of haptoglobin, EMT-associated motile phenotype was noted in OC2 cell lines. Overexpression of haptoglobin was also associated with an increased risk for locoregional recurrence [hazard ratio (HR) 1.04; p=0.011] after adjusting for age, gender, disease site, stage, and treatment modality. CONCLUSIONS: Increased cellular expression of haptoglobin is associated with EMT in oral cancer cell lines and this phenomenon could be exaggerated with IL-6. Cellular expression of haptoglobin is related to locoregional recurrence rate in buccal cancer patients.

Acceptable noise level (ANL) and real-world hearing-aid success in Taiwanese listeners.

OBJECTIVE: The objective of this prospective study was to investigate the relationship between acceptable noise level (ANL), which was measured using Taiwanese and the international speech test signal (ISTS), and real-world hearing-aid success for listeners who were representative of the population commonly seen in clinics. DESIGN: Unaided ANLs were measured pre-hearing-aid fitting. Hearing-aid success was assessed three months post-fitting using the international outcome inventory for hearing aids (IOI-HA) and a hearing-aid use questionnaire. STUDY SAMPLE: Eighty adults with hearing impairment completed the study. RESULTS: Both Taiwanese and ISTS ANLs were significantly associated with hearing-aid success, with higher ANLs suggesting poorer outcomes. However, the ANL's prediction accuracy for the probability of hearing-aid success was either much lower than that suggested by some literature, or was not much different from that of simply predicting all listeners as successful users. CONCLUSIONS: The current study suggested the possibility of using ANL to predict hearing-aid success. However, the usefulness of ANL as a clinical tool is unlikely to be as great as indicated by the literature.

The equivalence of acceptable noise level (ANL) with English, Mandarin, and non-semantic speech: a study across the U.S. and Taiwan.

OBJECTIVE: Acceptable noise level (ANL) determines the maximum noise level that a listener is willing to accept while listening to speech. The objective of this study was to determine the equivalence of ANL measured using different speech stimuli for native speakers who lived in the U.S. and Taiwan. DESIGN: ANLs were measured using English, Mandarin, and the international speech test signal (ISTS) at each site. The same babble noise was used across speech stimuli. The ANLs were considered equivalent if the difference was unlikely to be greater than 3 dB. STUDY SAMPLE: Thirty adults with normal hearing were recruited at each site. RESULTS: For each site, the equivalence test suggested that the native-language and foreign-language ANLs were equivalent. Between the two sites, ANLs measured using the listener's native language were also equivalent. Although the ISTS ANL obtained within each site was equivalent to, and highly correlated to, the native-language ANL, the data were unable to confirm the equivalence of the ISTS ANLs obtained from the two sites. CONCLUSIONS: The results suggested the possibility of directly comparing ANL measures carried out in different countries using different languages. However, it remains unclear if the ISTS can serve as an international ANL stimulus.

The nuclear receptor TLX (NR2E1) inhibits growth and progression of triple- negative breast cancer.

Development of targeted therapies will be a critical step towards reducing the mortality associated with triple-negative breast cancer (TNBC). To achieve this, we searched for targets that met three criteria: (1) pharmacologically targetable, (2) expressed in TNBC, and (3) expression is prognostic in TNBC patients. Since nuclear receptors have a well-defined ligand-binding domain and are thus highly amenable to small-molecule intervention, we focused on this class of protein. Our analysis identified TLX (NR2E1) as a candidate. Specifically, elevated tumoral TLX expression was associated with prolonged recurrence-free survival and overall survival for breast cancer patients with either estrogen receptor alpha (ERα)-negative or basal-like tumors. Using two TNBC cell lines, we found that stable overexpression of TLX impairs in vitro proliferation. RNA-Seq analysis revealed that TLX reduced the expression of genes implicated in epithelial-mesenchymal transition (EMT), a cellular program known to drive metastatic progression. Indeed, TLX overexpression significantly decreased cell migration and invasion, and robustly decreased the metastatic capacity of TNBC cells in murine models. We identify SERPINB2 as a likely mediator of these effects. Taken together, our work indicates that TLX impedes the progression of TNBC. Several ligands have been shown to regulate the transcriptional activity of TLX, providing a framework for the future development of this receptor for therapeutic intervention.

Function-adaptive clustered nanoparticles reverse Streptococcus mutans dental biofilm and maintain microbiota balance.

Dental plaques are biofilms that cause dental caries by demineralization with acidogenic bacteria. These bacteria reside inside a protective sheath which makes any curative treatment challenging. We propose an antibiotic-free strategy to disrupt the biofilm by engineered clustered carbon dot nanoparticles that function in the acidic environment of the biofilms. In vitro and ex vivo studies on the mature biofilms of Streptococcus mutans revealed >90% biofilm inhibition associated with the contact-mediated interaction of nanoparticles with the bacterial membrane, excessive reactive oxygen species generation, and DNA fragmentation. An in vivo examination showed that these nanoparticles could effectively suppress the growth of S. mutans. Importantly, 16S rRNA analysis of the dental microbiota showed that the diversity and richness of bacterial species did not substantially change with nanoparticle treatment. Overall, this study presents a safe and effective approach to decrease the dental biofilm formation without disrupting the ecological balance of the oral cavity.

Sylvatic Canine Morbillivirus in Captive Panthera Highlights Viral Promiscuity and the Need for Better Prevention Strategies.

Canine Distemper Virus (CDV) is a multi-host morbillivirus that infects virtually all Carnivora and a few non-human primates. Here we describe a CDV outbreak in an exotic felid rescue center that led to the death of eight felids in the genus Panthera. Similar to domestic dogs and in contrast to previously described CDV cases in Panthera, severe pneumonia was the primary lesion and no viral antigens or CDV-like lesions were detected in the central nervous system. Four tigers succumbed to opportunistic infections. Viral hemagglutinin (H)-gene sequence was up to 99% similar to strains circulating contemporaneously in regional wildlife. CDV lesions in raccoons and skunk were primarily encephalitis. A few affected felids had at least one previous vaccination for CDV, while most felids at the center were vaccinated during the outbreak. Panthera sharing a fence or enclosure with infected conspecifics had significantly higher chances of getting sick or dying, suggesting tiger-tiger spread was more likely than recurrent spillover. Prior vaccination was incomplete and likely not protective. This outbreak highlights the need for further understanding of CDV epidemiology for species conservation and public health.

Treatment results and prognostic factors in locally advanced hypopharyngeal cancer.

CONCLUSIONS: We suggest that concurrent chemoradiation (CCRT) is an effective definitive treatment for patients with advanced hypopharyngeal carcinoma who are unfit for or refuse surgery. A high dose of radiation (> 70 Gy) should be given to achieve acceptable local control rates and survival. OBJECTIVES: The purpose of this retrospective study was to compare the treatment results of locally advanced hypopharyngeal carcinoma with two different protocols. PATIENTS AND METHODS: From December 1995 to December 2004, 74 patients with locally advanced hypopharyngeal cancer were treated with CCRT or surgery plus postoperative radiotherapy (SRT). Their treatment results were reviewed by retrospective analysis. The study points included outcome, toxicity, and prognostic factors. RESULTS: There was no significant difference in T and N status between the two treatment groups, nor were there significant differences in overall or disease-free survival or the incidence of distant metastasis (p >0.05). In the CCRT group and SRT group, the estimated 3-year overall survival was 39% and 44%, respectively. The SRT group had better local control than the CCRT group (p <0.05). Relatively, 27% patients retained their larynx function for more than 2 years in the CCRT group. Radiation doses >70 Gy yielded significantly better survival and local control than doses <70 Gy (p <0.05).

A preliminary result of concurrent chemoradiation with weekly cisplatin in elderly nasopharyngeal carcinoma patients.

CONCLUSION: A weekly regimen of cisplatin 30 mg/m2 as concurrent chemoradiation (CCRT) was effective in elderly nasopharyngeal carcinoma (NPC) patients, with acceptable and reversible acute toxicity following CCRT therapy. OBJECTIVES: The purpose of this retrospective study was to evaluate the efficacy, toxicity, and tolerability of a multi-modal treatment strategy in elderly NPC patients. Subsequent systemic adjuvant chemotherapy was administered to achieve systemic control. PATIENTS AND METHODS: From December 2002 to December 2006, 26 NPC patients over 60 years of age who had stage IIB to IV NPC were evaluated in this retrospective analysis. The CCRT chemotherapy protocol consisted of eight weekly doses of cisplatin 30 mg/m2 administered in an outpatient setting. Adjuvant systemic chemotherapy consisted of four monthly cycles of cisplatin (20 mg/m2/day) plus 5-fluorouracil (1000 mg/m2/day) for 5 consecutive days. Study end points included treatment outcome, compliance, and toxicity. RESULTS: The 2-year overall survival, disease-free survival, local control, and distant metastasis-free rate were 87%, 73%, 92%, and 76%, respectively. Over 80% of patients were able to take more than six doses of weekly cisplatin during CCRT; however, nearly half of the patients had grade 3 hematological toxicity during adjuvant therapy requiring treatment modification or cessation of further adjuvant therapy.

Prognostic influence of parapharyngeal extension in nasopharyngeal carcinoma.

CONCLUSIONS: Advanced parapharyngeal tumor involvement in nasopharyngeal carcinoma (NPC) had significant predictive value associated with poorer treatment outcome. Further subclassification of parapharyngeal invasion may be considered in the TNM staging system. OBJECTIVES: We conducted a retrospective study to elucidate the effect of parapharyngeal extension on treatment outcomes in patients with NPC who were treated with radiotherapy or concurrent chemoradiotherapy (CCRT). PATIENTS AND METHODS: A total of 99 patients with newly diagnosed NPC were enrolled in this study. The parapharyngeal space invasion examined by CT scan was identified and graded according to Sham and Choy's classification. Potentially significant parameters were analyzed by both univariate and multivariate methods using SPSS software. RESULTS: The overall survival, recurrence-free survival, locoregional control survival and distant metastasis-free survival rates were affected by the presence of parapharyngeal space involvement (p<0.0001, p<0.0001, p<0.0001, p=0.002, respectively). In multivariate analysis accounting for all previously known prognostic factors, parapharyngeal invasion was associated with increased risk for any recurrence, locoregional recurrence, distant metastasis, and overall survival. After adjusting with TNM classification, parapharyngeal invasion was still an independent prognostic factor in NPC.

Primary tumor volume of nasopharyngeal carcinoma: significance for survival.

OBJECTIVES: To elucidate the effect of primary tumor volume (PTV) on treatment outcomes in patients with nasopharyngeal carcinoma (NPC) who were treated with radiotherapy or concurrent chemoradiotherapy. The TNM staging system developed by American Joint Committee of Cancer (AJCC) is universally used and accepted but its prediction of prognosis in NPC receives a lot of challenge. Primary tumor volume had been reported to have close relationship with prognosis of head and neck cancer. We may predict prognosis of NPC with PTV. METHODS: From 1999 to 2006, 91 patients with newly diagnosed NPC who were treated with radiotherapy or CCRT were enrolled in the study. Computed tomography-derived or magnetic resonance-derived primary tumor volume was calculated. The correlation between AJCC disease stage, primary tumor volume and disease-specific survival were analyzed. Multivariate analyses using the Cox proportional hazard model was performed. RESULTS: The median primary tumor volume for the whole series was 11.39 ml (range 1.25-166.58 ml). The median primary tumor volume was 2.69 ml in T1 disease, 10.14 ml in T2 disease, 15.41 ml in T3 disease, and 26.69 ml in T4 disease. Hazard ratio increased with tumor volume, ranging from 5.91 (95% confidence interval (95% CI) 1.13-26.43) for tumor volumes between 20 ml and 40 ml, and 15.72 (95% CI, 3.82-61.05) for tumor volumes >40 ml. With both tumor volume and T classification in the same Cox regression model, only tumor volume remained statistically significant in the prognosis of NPC. CONCLUSION: Primary tumor volume has closer relationship with survival rates of patients with NPC. Calculation of primary tumor volume may be further considered to improve the current staging system.

Use of formalin-fixed tissues to determine fumonisin B1-induced sphingolipid alterations in swine.

Fumonisin B1 is a mycotoxin that causes lethal pulmonary edema in swine. Sphinganine, sphingosine, and the sphinganine to sphingosine ratio are important biomarkers for fumonisin B1 exposure. Currently, tissues selected for sphinganine and sphingosine analyses are frozen at -80 degrees C until analyses take place. However, for diagnostics and some research projects, formalin is used more routinely as a preservative for long-term storage of tissues. To determine whether formalin-fixed tissues could be used for sphinganine and sphingosine analyses, sphinganine and sphingosine concentrations were quantified in both frozen and formalin-fixed lung, liver, kidney, and heart from fumonisin B1-treated and control pigs. Tissues were evaluated 3 months after freezing and 3, 6, and 12 months after formalin fixation. Sphinganine, sphingosine, and the sphinganine to sphingosine ratio of both frozen and formalin-fixed lung and liver from fumonisin B1-treated pigs were elevated. Formalin-fixed tissues had lower sphinganine and sphingosine concentrations but higher sphinganine to sphingosine ratios than the corresponding frozen tissues. Storage in formalin for up to 12 months did not affect the results. Sphingosine analysis could not be performed in formalin-fixed heart and kidney because of noninterpretable chromatograms. Therefore, formalin-fixed lung and liver can be used to determine fumonisin B1-induced sphinganine and sphingosine alterations in swine, with the sphinganine to sphingosine ratio being the most useful.