A typical enzyme activity for glutathione conjugation indicates exposure of pacu to pollutants

Functional enzyme assays to detect sublethal poisoning of Neotropical fish are paramount. Accordingly, we assayed a glutathione S-transferase (GST) activity in liver and kidney cytosols from Piaractus mesopotamicus injected with methyl parathion or benzo[a]pyrene using the substrate 1-chloro-2,4-dinitrobenzene (CDNB), which is the usual substrate for assaying a known general activity of GST. Since the most reactive substrate is required to reveal specific changes in enzyme activity, we also used two alternative substrates, 1,2-dichloro-4-nitrobenzene (DCNB) and ethacrynic acid (ETHA). Activities with CDNB or ETHA did not change. However, assays with DCNB showed that methyl parathion caused a decrease in GST activity in the liver on the 24th, 48th and 96th hour after the injection. DCNB also revealed that GST activity in the liver increased seven days after benzo[a]pyrene injection, coming down to normal after fourteen days. Benzo[a]pyrene, but not methyl parathion, increased the activities with DCNB in cytosol from the kidney seven and fourteen days after the injection. Thus, a decreased liver GST activity assayed with DCNB corresponded to contamination of P. mesopotamicus with methyl parathion. The increase of this GST activity in the liver and the kidney correlates to pacu contamination with polycyclic aromatic hydrocarbons.

Ensaios práticos de enzimas para detectar contaminação subletal de peixes neotropicais são da maior importância. Assim, ensaiamos a atividade da glutationa S-transferase (GST) em citosóis de fígado e de rim de Piaractus mesopotamicus injetados com metilparation ou benzo[a]pireno usando 1-cloro-2,4-dinitrobenzeno (CDNB), o substrato usual para ensaiar uma denominada atividade geral de GST. Desde que para indicar alterações na atividade de uma isoenzima, é necessário o substrato mais reativo, também usamos dois substratos alternativos, o 1,2-dicloro-4-nitrobenzeno (DCNB) e o ácido etacrínico (ETHA). As atividades com CDNB ou ETHA não mudaram. Entretanto, ensaios com DCNB mostraram que metilparation decresceu a atividade de GST no fígado em 24, 48 e 96 horas depois da injeção. O DCNB também revelou que sete dias depois da injeção de benzo[a]pireno a atividade da GST aumentou no fígado, normalizando depois de 14 dias. Benzo[a]pireno, mas não metilparation, aumentou as atividades com DCNB no citosol dos rins sete e 14 dias depois da injeção. Assim, uma atividade de GST ensaiada com DCNB diminuída no fígado correspondeu à contaminação de P. mesopotamicus com metilparation. O aumento dessa atividade de GST no fígado e nos rins está correlacionada com contaminação do pacu por hidrocarbonetos policíclicos aromáticos.

Infrared Emission from Photodissociation of Methyl Formate [HC(O)OCH3] at 248 and 193 nm: Absence of Roaming Signature.

Following photodissociation at 248 nm of gaseous methyl formate (HC(O)OCH3, 0.73 Torr) and Ar (0.14 Torr), temporally resolved vibration-rotational emission spectra of highly internally excited CO (ν ≤ 11, J ≤ 27) in the 1850-2250 cm-1 region were recorded with a step-scan Fourier-transform spectrometer. The vibration-rotational distribution of CO is almost Boltzmann, with a nascent average rotational energy (ER0) of 3 ± 1 kJ mol-1 and a vibrational energy (EV0) of 76 ± 9 kJ mol-1. With 3 Torr of Ar added to the system, the average vibrational energy was decreased to EV0 = 61 ± 7 kJ mol-1. We observed no distinct evidence of a bimodal rotational distribution for ν = 1 and 2, as reported previously [Lombardi et al., J. Phys. Chem. A 2016, 129, 5155], as evidence of a roaming mechanism. The vibrational distribution with a temperature of ∼13000 ± 1000 K, however, agrees satisfactorily with trajectory calculations of these authors, who took into account conical intersections from the S1 state. Highly internally excited CH3OH that is expected to be produced from a roaming mechanism was unobserved. Following photodissociation at 193 nm of gaseous HC(O)OCH3 (0.42 Torr) and Ar (0.09 Torr), vibration-rotational emission spectra of CO (ν ≤ 4, J ≤ 38) and CO2 (with two components of varied internal distributions) were observed, indicating that new channels are open. Quantum-chemical calculations, computed at varied levels of theory, on the ground electronic potential-energy schemes provide a possible explanation for some of our observations. At 193 nm, the CO2 was produced from secondary dissociation of the products HC(O)O and CH3OCO, and CO was produced primarily from secondary dissociation of the product HCO produced on the S1 surface or the decomposition to CH3OH + CO on the S0 surface.

Complete mitochondrial genome sequences of Atlantic representatives of the invasive Pacific coral species Tubastraea coccinea and T. tagusensis (Scleractinia, Dendrophylliidae): Implications for species identification.

Members of the azooxanthellate coral genus Tubastraea are invasive species with particular concern because they have become established and are fierce competitors in the invaded areas in many parts of the world. Pacific Tubastraea species are spreading fast throughout the Atlantic Ocean, occupying over 95% of the available substrate in some areas and out-competing native endemic species. Approximately half of all known coral species are azooxanthellate but these are seriously under-represented compared to zooxanthellate corals in terms of the availability of mitochondrial (mt) genome data. In the present study, the complete mt DNA sequences of Atlantic individuals of the invasive scleractinian species Tubastraea coccinea and Tubastraea tagusensis were determined and compared to the GenBank reference sequence available for a Pacific "T. coccinea" individual. At 19,094bp (compared to 19,070bp for the GenBank specimen), the mt genomes assembled for the Atlantic T. coccinea and T. tagusensis were among the longest sequence determined to date for "Complex" scleractinians. Comparisons of genomes data showed that the "T. coccinea" sequence deposited on GenBank was more closely related to that from Dendrophyllia arbuscula than to the Atlantic Tubastraea spp., in terms of genome length and base pair similarities. This was confirmed by phylogenetic analysis, suggesting that the former was misidentified and might actually be a member from the genus Dendrophyllia. In addition, although in general the COX1 locus has a slow evolutionary rate in Scleractinia, it was the most variable region of the Tubastraea mt genome and can be used as markers for genus or species identification. Given the limited data available for azooxanthellate corals, the results presented here represent an important contribution to our understanding of phylogenetic relationships and the evolutionary history of the Scleractinia.

Two New Species of the Genus Criotettix Bolivar (Orthoptera: Tetrigidae), with a Key to the Species of the Genus from China.

Two new species of the tetrigid genus Criotettix Bolivar from China, namely Criotettix interruptaoides Deng & Zheng n. sp. and Criotettix shiwanshanensis Deng & Zheng n. sp. are described. An updated key to all known species of the genus from China is given.

Comparison of extraction methods of total microbial DNA from freshwater.

The demand for molecular analysis of aquatic microbial communities in freshwater has highlighted the need for efficient methods of DNA extraction. The centrifugation method and filtration-membrane method are 2 widely used methods for extracting DNA. The objective of this study was to compare the extraction efficiency of 3 methods, including the centrifugation method, filtration-membrane method, and modified filtration-membrane method, by evaluating the quantity and purity of DNA extracts obtained from water. DNA extraction was analyzed by agarose gel electrophoresis, ultraviolet-spectroscopy, restriction enzyme digestion, and polymerase chain reaction. The results showed that the modified filtration-membrane method was the most efficient for extracting microbial DNA from freshwater with high integrity and purity and is suitable for molecular applications.

A forced running wheel system with a microcontroller that provides high-intensity exercise training in an animal ischemic stroke model

We developed a forced non-electric-shock running wheel (FNESRW) system that provides rats with high-intensity exercise training using automatic exercise training patterns that are controlled by a microcontroller. The proposed system successfully makes a breakthrough in the traditional motorized running wheel to allow rats to perform high-intensity training and to enable comparisons with the treadmill at the same exercise intensity without any electric shock. A polyvinyl chloride runway with a rough rubber surface was coated on the periphery of the wheel so as to permit automatic acceleration training, and which allowed the rats to run consistently at high speeds (30 m/min for 1 h). An animal ischemic stroke model was used to validate the proposed system. FNESRW, treadmill, control, and sham groups were studied. The FNESRW and treadmill groups underwent 3 weeks of endurance running training. After 3 weeks, the experiments of middle cerebral artery occlusion, the modified neurological severity score (mNSS), an inclined plane test, and triphenyltetrazolium chloride were performed to evaluate the effectiveness of the proposed platform. The proposed platform showed that enhancement of motor function, mNSS, and infarct volumes was significantly stronger in the FNESRW group than the control group (P<0.05) and similar to the treadmill group. The experimental data demonstrated that the proposed platform can be applied to test the benefit of exercise-preconditioning-induced neuroprotection using the animal stroke model. Additional advantages of the FNESRW system include stand-alone capability, independence of subjective human adjustment, and ease of use.

A forced running wheel system with a microcontroller that provides high-intensity exercise training in an animal ischemic stroke model.

We developed a forced non-electric-shock running wheel (FNESRW) system that provides rats with high-intensity exercise training using automatic exercise training patterns that are controlled by a microcontroller. The proposed system successfully makes a breakthrough in the traditional motorized running wheel to allow rats to perform high-intensity training and to enable comparisons with the treadmill at the same exercise intensity without any electric shock. A polyvinyl chloride runway with a rough rubber surface was coated on the periphery of the wheel so as to permit automatic acceleration training, and which allowed the rats to run consistently at high speeds (30 m/min for 1 h). An animal ischemic stroke model was used to validate the proposed system. FNESRW, treadmill, control, and sham groups were studied. The FNESRW and treadmill groups underwent 3 weeks of endurance running training. After 3 weeks, the experiments of middle cerebral artery occlusion, the modified neurological severity score (mNSS), an inclined plane test, and triphenyltetrazolium chloride were performed to evaluate the effectiveness of the proposed platform. The proposed platform showed that enhancement of motor function, mNSS, and infarct volumes was significantly stronger in the FNESRW group than the control group (P<0.05) and similar to the treadmill group. The experimental data demonstrated that the proposed platform can be applied to test the benefit of exercise-preconditioning-induced neuroprotection using the animal stroke model. Additional advantages of the FNESRW system include stand-alone capability, independence of subjective human adjustment, and ease of use.

Expression of RUNX3 gene in pancreatic adenocarcinoma and its clinical significance.

We investigated the clinical significance of RUNX3 gene expression in human pancreatic carcinoma. Five samples of pancreatic tissues and 30 samples of pancreatic cancer tissues and paracancerous tissues were collected. RUNX3 expression was detected by real-time PCR and immunohistochemistry. The relationships between clinicopathological findings and the expression of RUNX3 were analyzed. The relative quantification level of RUNX3 mRNA expression in human pancreatic carcinoma tissues and paracancerous tissues was 2.60 (0.42-12.82) and 1.02 (0.19-3.58), respectively (P < 0.05). The percentage of positive cells expressing RUNX3 protein in human pancreatic tissues and paracancerous tissues was 45.5 ± 26.2 and 6.9 ± 6.0%, respectively (P < 0.01). The high RUNX3 group (N = 9) with 45.5% or more of the cancer cells staining for RUNX3 and the low RUNX3 group (N = 21) with less than 45.5% cancer cells staining for RUNX3. Low expression of RUNX3 correlated significantly with an advanced TNM stage (χ(2) = 6.897, P = 0.045), lymph node metastasis (χ(2) = 4.739, P = 0.029) and neural invasion (χ(2) = 5.44, P = 0.020). On the other hand, no association could be found between RUNX3 expression and clinicopathological variables including age, gender, tumor location, tumor size, tumor differentiation or the serum concentration of CEA and CA199. The expression of RUNX3 in pancreatic cancer tissues was obviously higher than that in the paracancerous tissues. Low expression of RUNX3 may have an important role in aggressiveness, lymph node metastasis and neural invasion in pancreatic cancer. In pancreatic carcinoma tissues, low expression of RUNX3 may indicate a poor prognosis.

Diagnostic value of cytological and microbiological methods in cryptococcal meningitis.

The aim of this study was to investigate diagnostic methods for cryptococcal meningitis (CM). A retrospective analysis was conducted for 31 patients with CM confirmed by etiologic detection of cerebrospinal fluid in our hospital in the past 5 years. Nineteen cases in 31 patients were confirmed with CM in the first diagnosis, with a misdiagnosis rate of 38.7%. The positive rates of cryptococcus detection in cerebrospinal fluid with May-Grünwald-Giemsa (MGG)-, ink-, and Alcian blue-staining methods were 86.9, 70.9, and 80.6%, respectively. The misdiagnosis rate of CM is high during the early stage of disease. The total positive rate of cryptococcus diagnosis using the MGG-staining method was significantly higher than that using the ink-staining method. These results are important for diagnosing CM.

Two New Species of the Genus Xistra Bolivar (Orthoptera: Tetrigidae) from China.

Two new species of the tetrigid genus Xistra Bolivar from China, namely Xistra oculata Li, Deng et Zheng n. sp. and Xistra brachynota Li, Deng et Zheng n. sp. are described. An updated key to all known species of the genus is given.

Description of a New Species of the Genus Macromotettixoides Zheng (Orthoptera: Tetrigoidea: Metrodorinae) from China.

A new species, Macromotettixoides lativertex Deng et al n. sp. is described with detailed illustrations of external morphology. Comparison between the species M. lativertex n. sp. and allied species is compiled into a table for identification. The distribution and an updated identification key to all known species of the genus is given.

WE-G-BRB-09: CyberKnife Patient Specific QA Using a 4D Cylindrical Diode Array System.

PURPOSE: To examine and facilitate the feasibility of the ArcCheck cylindrical diode array system as a patient specific QA device for CyberKnife radiosurgery delivery. METHODS: There is an obvious necessity for CyberKnife robotic radiosurgery patient QA procedures for hypofractionated treatment of larger planned treatment volumes (PTV), e.g. prostate. This need will increase when the future CyberKnife MLC is introduced. The small unflattened CyberKnife fields, along with the variation of beam-to-detector spatial angles, pose a significant detection challenge for dosimetric systems. The feasibility of the ArcCheck (Sun Nuclear Inc.) cylindrical diode array system for patient-specific QA on the CyberKnife is demonstrated using a beam-to-diode specific angular correction that was developed and has been applied. For localization and tracking, four gold seed fiducial markers were embedded in the system's central plug. We used a Monte Carlo 1% uncertainty for the dose calculation. RESULTS: By disabling the Linac based corrections and applying the custom CyberKnife correction that we developed, the passing rate increased from 39.6% to 99.8% using a 3%3mm gamma criteria for a given lung case. An additional lung case passed 98.5%. In both cases, a 10% dose threshold was used. In addition, brain, trigeminal nerve and lung cases with synchrony tracking are being investigated. CONCLUSIONS: We demonstrated the ArcCheck feasibility for CyberKnife patient specific QA performance. The custom CK angular correction that we developed and applied showed a high passing rate for the lung cases. A verification of the polar angle response should be conducted, in addition to the azimuthal angle that was verified for Linacs. Any data that is being retrieved is additional data to the current chamber point measurement procedures.

SU-E-T-383: Pulsed Low Dose Rate Radiotherapy Using Advanced Treatment Methods: A Novel Technique for Patient Re-Irradiation.

PURPOSE: Pulsed low dose-rate radiotherapy (PLDR) re-irradiation has the potential to reduce late normal tissue toxicity while still providing significant tumor control for recurrent cancers. In contrast to conventional treatments delivered at dose-rates of 400-600cGy/min, PLDR treatments deliver 20cGy pulses separated by 3-minute intervals to achieve an effective-dose-rate of 6.7cGy/min. This work aims to investigate the planning strategy and delivery quality of PLDR treatment using IMRT and RapidArc techniques. METHODS: Twenty cases (10 treated with PLDR IMRT, 10 for evaluation purposes) were recruited in this study including prostate, pancreas, lung, head-and-neck, breast and pelvis. IMRT and the RapidArc treatment plans were generated using the Eclipse TPS. For IMRT treatment, each plan consisted of 10 fields to achieve a daily dose of 200cGy. The breast IMRT and the RapidArc plans consisted of two fields/arcs, respectively (40cGy/plan) and were delivered 5 times. The dose contribution from each field to the planning target volume (PTV) was analyzed to evaluate the feasibility for PLDR treatment. Machine-operation- dose-rate and plan quality was also investigated. Dose delivery accuracy was assessed using a cylindrical diode array. RESULTS: Throughout the six treatment sites, the mean PTV dose ranged from 16.1 to 26.1cGy/arc for RapidArc plans and 10.3 to 36.7cGy/field for IMRT plans. For IMRT, the PTV dose contribution from each field strongly depends on the beam arrangement and optimization parameters. With very low dose for a full rotation (∼ 20cGy/arc), the machine-operation-dose-rate of RapidArc plans significantly affects plan quality and deliverability. A machine-operation-dose-rate of 100 MU/min results in superior delivery accuracy (>97.7% gamma-passing-rate for 3%/3mm criteria) for both IMRT and RapidArc plans. CONCLUSIONS: PLDR radiotherapy using IMRT and RapidArc techniques Result in both dosimetric and radiobiological benefits, which may have great potential for those previously-irradiated patients who have historically done poorly.

SU-E-T-464: Impact of the Treatment Margin on Tumor Control and Normal Tissue Complication for Prostate Treatment.

PURPOSE: To investigate the consequence of treatment margin reduction on normal tissue complication probability (NTCP) and tumor control probability (TCP) of prostate external beam treatment. METHODS: Intensity modulated rotational radiotherapy plans were generated for 10 prostate patients with 6 different posterior margin sizes from 5mm to 0. The prescription dose is 80Gy for 40 treatment fractions. The dose distributions were recalculated with consideration of the intrafractional motion and the localization error. The statistical uncertainties of the intrafractional motion and the localization error were derived based on the motion tracking data recorded by the Calypso 4D localization system for a large patient population. The TCP and NTCP were calculated based on the dose volume histograms (DVH) of prostate and rectum for plans with different margins using an equivalent uniform dose (EUD) based biological model. The 50% tumor control dose (TCD50) of 60Gy for prostate and the median toxic dose (TD50) of 55Gy for rectum were used in the calculation. RESULTS: The minimum dose of the prostate and the mean dose of the rectum dropped with the decrease of the treatment margin. When the posterior treatment margin was reduced from 5mm to zero, the EUD of prostate decreased from 83Gy (±0.5Gy) to 81Gy (±0.5Gy) and the TCP dropped from 93.2% (±0.1%) to 91.7% (±0.1%), the EUD of the rectum decreased more significantly from 48.9Gy (±0.4Gy) to 32.5Gy (±0.5Gy) and the NTCP dropped from 13.3% (±1.5%) to 0.03% (± 0.01%). CONCLUSIONS: The treatment margin size affects the dose to the target and the nearby critical structure. More significant impact on NTCP has been observed than on TCP. This gives us some room to consider the quality of the patient's after-treatment life. A wise choice of treatment margin can be made based on physician's opinion and patient's preference on the tumor control and the quality of life.

TU-E-BRB-06: Best in Physics (Therapy) - Development and Experimental Validation of EPID-Based 4D Dose Reconstruction.

PURPOSE: To develop and validate an EPID-based 4D patient dose reconstruction framework accounting for linac delivery uncertainties, interfractional and intrafractional motions, and interplay effect. METHODS: Patients with fiducial markers were scanned with 4D-CT for SBRT planning. Before treatment, in-room 4D-CT was performed. Both the MLC and the tumor movements were tracked by continuously acquiring EPID images during treatment. Instead of directly using the heterogeneous transit photon fluence measured by the EPID, this method reconstructed the incident beam fluence based on the MLC apertures measured by the EPID and the delivered MU recorded by the linac. To account for the time-dependent-geometry, the incident fluence distributions were sorted into their corresponding phases based on the tumor motion pattern detected by the EPID and accumulated as the incident fluence map for each phase. Together with 4D-CT, it was then used for Monte Carlo dose calculation. Deformable registration was performed to sum up the phase doses for treatment assessment. The feasibility of using the transit EPID images for incident fluence reconstruction was evaluated against EPID in-air measurements. The accuracy of 3D- and 4D-dose reconstruction was validated by a motordriven cylindrical diode array for six clinical SBRT plans. RESULTS: The average difference between the measured and reconstructed fluence maps is within 0.16%. The reconstructed 3D-dose shows 1.4% agreement in the CAX-dose and >98.5% gamma-passing-rate (2%/2mm) in the peripheral-dose. A distorted dose distribution is observed in the measurement for the moving ArcCheck-phantom. The comparison between the measured and the reconstructed 4D-dose without considering interplay fails the gammaevaluation (59%-88.9% gamma-passing-rate). In contrast, when the interplay is considered, the dose distortion phenomena is successfully represented in the reconstructed dose (>97.6% gamma-passing-rate). CONCLUSIONS: The experimental validation demonstrates that the proposed method provides a practical way to reconstruct the fractional 4D-doses received by the patient and enables adaptive SBRT strategy.

Carnitine and carnitine orotate affect the expression of the prolactin-releasing peptide gene.

Carnitine is involved in fatty acid metabolism in mammals and is widely used as a nutritional supplement; carnitine orotate is a more absorbable form of carnitine. We investigated the effects of carnitine and carnitine orotate on mouse prolactin-releasing peptide (PrRP) mRNA expression. Twenty-four female mice were randomly divided into four groups of six; control mice were orally drenched with physiological saline solution (250 mg/kg body weight) and treatment mice were orally drenched with carnitine (250 mg/kg) or carnitine orotate (250 or 750 mg/kg), once a day, for 20 days from parturition. The carnitine or carnitine orotate was dissolved in saline solution before administration. The hypothalamus, pituitary and ovary were sampled on day 21 after parturition, and PrRP mRNA levels in these tissues were measured by semi-quantitative PCR, with glyceraldehyde 3-phosphate dehydrogenase as a control. Expression of PrRP in mice treated with carnitine and carnitine orotate was significantly increased in the ovary and significantly reduced in the pituitary gland. Compared with the control, hypothalamus PrRP mRNA increased significantly in the carnitine and low-dose carnitine orotate groups and decreased significantly in the high-dose carnitine orotate group. We conclude that carnitine and carnitine orotate regulate expression of PrRP in the pituitary gland and ovaries.

Polymorphisms within the tumor necrosis factor and lymphotoxin-alpha genes and endemic pemphigus foliaceus--are there any associations?

The purpose of this study was to analyze the possible influence of the TNF and LTA loci polymorphisms on the susceptibility/resistance to endemic pemphigus foliaceus, also named fogo selvagem (FS), an autoimmune disease characterized by blisters due to acantholysis of the superficial-most epidermal cells. Autoantibodies, mainly of the IgG4 subclass, are directed against a desmosomal glycoprotein known as desmoglein 1. FS shares clinical, histological and immunological features with nonendemic pemphigus foliaceus. Most residents of the endemic regions do not develop the disease, and familial clustering has been documented, suggesting that host factors play a role in susceptibility. In fact, strong positive and negative associations with HLA class II genes have been reported. The TNF and LTA genes are located in the class III region of the Human Major Histocompatibility Complex. Their location, the function of their products, which are cytokines and pluripotent immunomodulators, as well as their genetic variability make them candidate genes for complex diseases with an altered immune response. A total of 162 patients and 191 controls were enrolled in this study. No significant associations were found with any one of the three LTA single nucleotide polymorphisms (SNP) analyzed (at nucleotides 249, 365, 720), nor with the TNF SNP located at positions -863 and -308. The frequency of allele TNF*238A was slightly decreased in patients (OR = 0.45). In conclusion, the results of this study indicate that genetic variability of the TNF and LTA genes does not play a major role in susceptibility/resistance to pemphigus foliaceus.

The prevalence of antibodies against desmoglein 1 in endemic pemphigus foliaceus in Brazil. Cooperative Group on Fogo Selvagem Research.

BACKGROUND: Pemphigus foliaceus is an autoimmune skin disease mediated by autoantibodies against desmoglein 1. The endemic form is thought to have an environmental cause. The Terena reservation of Limão Verde in Mato Grosso do Sul, Brazil, is a recently identified focus of the disease, with a prevalence of 3.4 percent in the population. We tested the hypothesis that normal subjects living in an endemic area have antibodies against desmoglein 1. METHODS: We used an enzyme-linked immunosorbent assay to detect antibodies against desmoglein 1 in serum samples from 60 patients with endemic pemphigus foliaceus (fogo selvagem) who lived in Limão Verde or elsewhere in Brazil, 372 normal subjects (without pemphigus foliaceus) from Limão Verde and surrounding locations, and 126 normal subjects from the United States and Japan. RESULTS: Antibodies against desmoglein 1 were detected in 59 of the 60 patients with fogo selvagem (98 percent) but in only 3 of the 126 normal subjects from the United States and Japan (2 percent). Antibodies were also detected in 51 of the 93 normal subjects from Limão Verde (55 percent) and in 54 of the 279 normal subjects from surrounding areas (19 percent). Serum samples obtained one to four years before the onset of disease were available for five patients; all five had antibodies in the initial serum samples, and the onset of disease was associated with a marked increase in antibody values. CONCLUSIONS: The prevalence of antibodies against desmoglein 1 is high among normal subjects living in an area among where fogo selvagem is endemic, and the onset of the disease is preceded by a sustained antibody response. These findings support the concept that the production of antibodies against desmoglein 1 is initiated by exposure to an unknown environmental agent.