Frequency and characterization of the accessory heads of the biceps brachii: A multicentric cadaveric study.

PURPOSE: The biceps brachii muscle (BBM) is a large and thick muscle on the ventral portion of the upper arm. The scientific literature reports the existence of extra heads. OBJECTIVE: The present work investigated the frequency of occurrence of BBM accessory head(s) in four different Anatomy Laboratories at universities in Northeast Brazil. MATERIAL AND METHODS: Once the upper limbs with an anatomical variation of the BBM heads were identified, the accessory head was evaluated for morphometric characterization (the length, width, and thickness). RESULTS: The sample consisted of 249 upper limbs and the outcomes were 26 members with BBM accessory heads (10.44% of the total sample). Among the 26 members studied was found a single accessory head in 22 (84.62%), two accessory heads in 3 (11.54%), and three accessory heads in 1 (3.85%). BBMs with one, two, and three accessory heads were found, with different origins. The average length, width, and thickness of these accessory heads were 169.39mm, 10.25mm, and 3.39mm, respectively. CONCLUSION: The frequency of BBM accessory heads in a population of northeastern Brazil in this cadaveric study was 10.44%.

Temporal expression of pluripotency-associated transcription factors in sheep and cattle preimplantation embryos.

SummaryPluripotency-associated transcription factors (PATFs) modulate gene expression during early mammalian embryogenesis. Despite a strong understanding of PATFs during mouse embryogenesis, limited progress has been made in ruminants. This work aimed to describe the temporal expression of eight PATFs during both sheep and cattle preimplantation development. Transcript availability of PATFs was evaluated by reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) in eggs, cleavage-stage embryos, morulae, and blastocysts. Transcripts of five genes were detected in all developmental stages of both species (KLF5, OCT4, RONIN, ZFP281, and ZFX). Furthermore, CMYC was detected in all cattle samples but was found from cleavage-stage onwards in sheep. In contrast, NR0B1 was detected in all sheep samples but was not detected in cattle morulae. GLIS1 displayed the most significant variation in temporal expression between species, as this PATF was only detected in cattle eggs and sheep cleavage-stage embryos and blastocysts. In silico analysis suggested that cattle and sheep PATFs share similar size, isometric point and molecular weight. A phenetic analysis showed two patterns of PATF clustering between cattle and sheep, among several mammalian species. In conclusion, the temporal expression of pluripotency-associated transcription factors differs between sheep and cattle, suggesting species-specific regulation during preimplantation development.

The transcriptome of the mosquito Aedes fluviatilis (Diptera: Culicidae), and transcriptional changes associated with its native Wolbachia infection.

BACKGROUND: Wolbachia is a bacterial endosymbiont that naturally infects a wide range of insect species, and causes drastic changes to host biology. Stable infections of Wolbachia in mosquitoes can inhibit infection with medically important pathogens such as dengue virus and malaria-causing Plasmodium parasites. However, some native Wolbachia strains can enhance infection with certain pathogens, as is the case for the mosquito Aedes fluviatilis, where infection with Plasmodium gallinaceum is enhanced by the native wFlu Wolbachia strain. To better understand the biological interactions between mosquitoes and native Wolbachia infections, and to investigate the process of pathogen enhancement, we used RNA-Seq to generate the transcriptome of Ae. fluviatilis with and without Wolbachia infection. RESULTS: In total, we generated 22,280,160 Illumina paired-end reads from Wolbachia-infected and uninfected mosquitoes, and used these to make a de novo transcriptome assembly, resulting in 58,013 contigs with a median sequence length of 443 bp and an N50 of 2454 bp. Contigs were annotated through local alignments using BlastX, and associated with both gene ontology and KEGG orthology terms. Through baySeq, we identified 159 contigs that were significantly upregulated due to Wolbachia infection, and 98 that were downregulated. Critically, we saw no changes to Toll or IMD immune gene transcription, but did see evidence that wFlu infection altered the expression of several bacterial recognition genes, and immune-related genes that could influence Plasmodium infection. wFlu infection also had a widespread effect on a number of host physiological processes including protein, carbohydrate and lipid metabolism, and oxidative stress. We then compared our data set with transcriptomic data for other Wolbachia infections in Aedes aegypti, and identified a core set of 15 gene groups associated with Wolbachia infection in mosquitoes. CONCLUSIONS: While the scale of transcriptional changes associated with wFlu infection might be small, the scope is rather large, which confirms that native Wolbachia infections maintain intricate molecular relationships with their mosquito hosts even after lengthy periods of co-evolution. We have also identified several potential means through which wFlu infection might influence Plasmodium infection in Ae. fluviatilis, and these genes should form the basis of future investigation into the enhancement of Plasmodium by Wolbachia.

Development and characterization of new microsatellites for Eugenia dysenterica DC (Myrtaceae).

Microsatellite markers were developed for population genetic analyses of the Neotropical tree Eugenia dysenterica DC (Myrtaceae), after construction of a shotgun genomic library for microsatellite discovery. Nine primers were designed, of which 5 yielded amplified product. These primers were polymorphic for 97 individuals collected in 3 distinct localities. The number of alleles per locus (primer) ranged from 3 to 11 and expected heterozygosities varied from 0.309 to 0.884. The probability of locus identity was ~1.88 x 10(-4) and the probability of paternity exclusion was ~0.9367. The 5 microsatellite primer pairs may be suitable for population genetic studies such as parentage and fine-scale genetic analyses of this species.

Effect of dentofacial deformity on maximum isometric tongue strength.

The objective of this study was to determine whether dentofacial deformities (class II and class III) influence maximum isometric tongue strength compared with a group without deformities. A total of 126 adult patients participated in the study. Of these, 45 had a class II diagnosis (14 men and 31 women) and 81 a class III diagnosis (35 men and 46 women), all of them with indication of orthognathic surgery. Fifty adult volunteers (17 men and 33 women) with no changes in dental occlusion (class I) and with no clinical signs of dysfunction of the temporomandibular joint represented the control group. Tongue strength (in N) was measured with a dynamometer. The maximum strength of the anterior portion was determined with the instrument positioned on the retroincisor region and the maximum strength of the dorsum with the instrument positioned in the region of the hard palate. Data were analysed statistically by analysis of variance (anova) and by the Pearson correlation test. No significant difference (P > 0·05) in tongue strength in the anterior or dorsal region was observed between the groups with dentofacial deformities or between these groups and the control. The tongue strength of the anterior and dorsal regions was not influenced by the dentofacial deformity.

Comparing resistome profiles from anthropogenically impacted and non-impacted areas of two South Shetland Islands - Maritime Antarctica.

Whalers Bay, in Deception Island, has one of the most anthropogenically impacted areas in Maritime Antarctica. However, considering the volcanic nature (high concentrations of heavy metals) of Deception Island's soils, this putative anthropogenic impact should be carefully investigated. In this context, the objective of this study was to compare resistome profiles of impacted and non-impacted areas in Deception Island (Whalers Bay, Crater Lake, and Fumarole Bay) and Livingston Island (Hannah Point) in order to investigate the microbiome tolerance/resistance mechanisms selected as a function of environmental drivers. Metagenomics was used to search for genes conferring resistance/tolerance to antibiotics, biocides, and heavy metals. Whalers Bay has a greater diversity of antibiotic, biocide, and heavy metal resistance classes found in its microbiomes. However, Hannah Point, at Livingston Island, has a greater abundance of antibiotic and biocide resistance/tolerance genes. The microbiome of Deception Island's non-impacted areas (Crater Lake and Fumarole Bay) showed resistance/tolerance genes almost entirely to heavy metals. Pb was found in higher concentrations in Whalers Bay soil in comparison to the other areas, indicating human contamination. The non-metric multidimensional scaling (NMDS) analysis revealed that Pb concentrations influenced resistome profiles in Whalers Bay soil. Despite the effect of Pb on the microbial communities of Whalers Bay, most heavy metal concentrations did not have a significant impact on resistome genes, suggesting that the volcanic soil heavy metal concentration of Deception Island has little biological influence.

Tongue strength of healthy young adults.

The measurement of tongue strength contributes to the study of oro-facial physiology. The objective of this study was to investigate the influence of gender on tongue strength in young adults and to determine the differences in this strength between tongue regions. This study was conducted on 50 healthy volunteers (17 men and 33 women) with a mean age of 23 years, with no oro-facial myofunctional alterations. The strength of the anterior portion and of the dorsum of the tongue was analysed with a calibrated electronic dynamometer. The subjects were asked to apply maximum force. Men showed higher tongue strength values than women both in the anterior portion and in the dorsum of the tongue (P<0·05), and the strength of the anterior portion of the tongue was lower than that of the dorsum in both genders (P<0·05). We conclude that gender and region influenced tongue strength.

Preventive therapy compliance in pediatric tuberculosis - A single center experience.

INTRODUCTION: Despite its importance, there are some barriers to patient compliance in preventive therapy (PT) of tuberculosis (TB). The purpose of this study was to evaluate the compliance to appointments, PT and follow-up in a pediatric population after TB exposure, followed in a single TB outpatient center, and the subsequent identification of compliance determinants. METHODS: Retrospective analysis of all pediatric patients who underwent PT in Gaia TB outpatient center from January 2015 to June 2016. Patients were divided into two groups: compliant and non-compliant, according to adherence to screening, visits and medication. The data collection was based on review of medical records. RESULTS: A total of 72 patients were enrolled, 33 (45.8%) on chemoprophylaxis and 39 (54.2%) on latent tuberculosis infection (LTBI) treatment. The majority of patients were compliant (63.9%, n=46). Non-compliance was found in 36.1% (n=26): in 12 patients to contact screening, in 11 patients to PT and 22 patients did not attend medical appointments in the first place. In 10 patients, non-compliance was related to social problems/family dysfunction (low socioeconomic status and parent's unemployment). After putting in place several strategies, such as telephone contact, activating social services and direct observation of therapy, a compliance of 98.6% was achieved. Isoniazid was the main drug used (91.7%), during 9 months for LBTI. CONCLUSION: PT compliance in TB can be challenging, probably related to the lack of risk perception and caregiver's reluctance to undergo a prolonged treatment to an asymptomatic condition. We conclude that implementing interventions can considerably improve treatment compliance and reduce the risk of future tuberculosis development. We emphasize the success in compliance to a 9 month regimen of isoniazid in the vast majority of patients with LTBI.

Magnetic studies of CoFe2O4/SiO2 aerogel and xerogel nanocomposites.

The evolution of the structural and magnetic properties of nanocomposites formed by cobalt ferrite particles dispersed in xerogel and aerogel silica matrices (CoFe2O4/SiO2) have been studied as a function of the temperature of preparation and the amount of ferrite dispersed in the matrix. Wet samples with different amounts of CoFe2O4 in SiO2 matrix were prepared by sol-gel process in monolithic form. Xerogel and aerogel samples were prepared by controlled and hypercritical drying, respectively, and heated at various temperatures between 300 and 1100 degrees C. Superparamagnetic behavior has been observed by magnetization studies at room temperature for xerogels prepared at low temperature. Aerogel samples showed significant superparamagnetic fractions for all thermal treatment temperatures as determined by Mössbauer spectroscopy. Magnetization of the nanocomposites at 10 KOe applied field varied from 1 to 19 emu/g and the coercivity from 90 to 2320 Oe, respectively, for the different morphologies and textures of the analyzed material. The results show that besides the magnetization and coercivity depend on crystallite size, parameters such as ferrite content, porosity and drying conditions greatly influence the nanocomposite magnetic behavior.

Degradation of trans-polyisoprene after root filling with thermoplasticized techniques.

AIM: To evaluate ex vivo degradation of gutta-percha following six thermoplastic obturation techniques. METHODOLOGY: Ninety human-extracted mandibular premolars were selected and divided randomly into nine groups for filling. Group 1: thermomechanical compaction for 3 s with Konne gutta-percha points (Konne Ind. e Com. de Mat. Odontol., Belo Horizonte, MG, Brazil); Group 2: thermomechanical compaction for 3 s with Dentsply TP gutta-percha points (Dentsply Indústria e Comércio Ltda, Petrópolis, R.J. Brazil); Group 3: thermomechanical compaction for 10 s with Konne; Group 4: thermomechanical compaction for 10 s with Dentsply TP; Group 5: warm vertical condensation using System B (EIE/Analytic, Richmond, WA, USA) with Konne; Group 6: warm vertical condensation using System B with Dentsply TP; Group 7: vertical condensation with Konne; Group 8: vertical condensation with Dentsply TP; Group 9: Microseal cone (Analytic Endodontics, Glendora, CA, USA). A further four groups were assessed without using teeth, Group 10: Microseal microflow (Analytic Endodontics); Group 11: Obtura (Obtura Corporation, Penton, MO, USA); Group 12: Obtura flow (Obtura Corporation); Group 13: Thermafil (Dentsply Maillefer, Tulsa, OK, USA). The filling material was removed from the root canal and trans-1,4-polyisoprene isolated by solubilization of the root filling remnants in chloroform followed by filtration and centrifugation. By gel permeation chromatography and infrared spectroscopy, the occurrence and degree of degradation were assessed. The results were analysed statistically using the Kruskal-Wallis test. With differential scanning calorimetry, the thermal behaviour of the gutta-percha was determined. RESULTS: A significant decrease in polymer molar mass and the production of carboxyl and hydroxyl groups in the polymer were observed with thermomechanical compaction used for 10 s and vertical condensation filling techniques (P = 0.0001 and P = 0.0005, respectively). Other techniques caused no polymer degradation. CONCLUSION: Polyisoprene degrades with high temperature. Thermomechanical compaction for 10 s and vertical condensation were associated with the greatest degradative process.

A New Light Trap Model as an Alternative for Controlling Pests in Eucalyptus Plantations.

Eucalyptus plantations can be affected by species of defoliating caterpillars. The integrated management of this group primarily involves a monitoring system, natural enemies, and biological products. Alternative control methods, including the use of conventional light traps, have not been adopted, mostly because of their low efficiency. Therefore, a more efficient light trap model was developed. The new model allowed the capture of 3.6 times as many insects as the conventional model, with a 261% gain in control efficiency. The use of this new model represents another integrated management alternative for lepidopteran pests of eucalyptus plantations and other cultured plants.

New Radiation Dosimetry Estimates for [18F]FLT based on Voxelized Phantoms.

3'-Deoxy-3-[18F]fluorothymidine, or [18F]FLT, is a positron emission tomography (PET) tracer used in clinical studies for noninvasive assessment of proliferation activity in several types of cancer. Although the use of this PET tracer is expanding, to date, few studies concerning its dosimetry have been published. In this work, new [18F]FLT dosimetry estimates are determined for human and mice using Monte Carlo simulations. Modern voxelized male and female phantoms and [18F]FLT biokinetic data, both published by the ICRP, were used for simulations of human cases. For most human organs/tissues the absorbed doses were higher than those reported in ICRP Publication 128. An effective dose of 1.70E-02 mSv/MBq to the whole body was determined, which is 13.5% higher than the ICRP reference value. These new human dosimetry estimates obtained using more realistic human phantoms represent an advance in the knowledge of [18F]FLT dosimetry. In addition, mice biokinetic data were obtained experimentally. These data and a previously developed voxelized mouse phantom were used for simulations of animal cases. Concerning animal dosimetry, absorbed doses for organs/tissues ranged from 4.47 ± 0.75 to 155.74 ± 59.36 mGy/MBq. The obtained set of organ/tissue radiation doses for healthy Swiss mice is a useful tool for application in animal experiment design.

Resistance of mice to Leptospira infection and correlation with chemokine response.

Leptospirosis is globally widespread neglected disease, affecting most mammalian species. Clinical signs can be confused with other diseases which make the diagnosis and treatment difficult. Chemokines and cytokines are known for their role in the inflammatory and immune response to infections. The profile determination of chemokines' expressions in the course of infection may elucidate the defense mechanisms of the host and support the search for effective treatment strategies. We investigated the mechanisms of innate immunity through the comparison of chemokines induced during infection with L. interrogans in mice with different levels of susceptibility. We used lung and spleen tissues samples of mice from C3H/HeJ, C3H/HePas and Balb/c, respectively sensitive, intermediate susceptibility and resistant to the pathogen. The inoculation of L. interrogans in C3H/HeJ mice led a comparatively smaller change in chemokines expression in both spleen and lung tissues. In samples from spleens and lungs of C3H/HePas and Balb/c the higher increases occurred on CXCL9, CXCL16, CXCL5, CCL8 and CCL5 in Balb/c. Given the same genetic background, the differences in the responses of C3H/HePas compared to C3H/HeJ mice strongly suggest the role of chemokines for the survival of parental strain. Therefore, the greatest increase in CXC chemokines appears to be efficient to induce migration of cells to the secondary lymphoid organs and affected tissues, which is important to control infection. Overall, CXC chemokines are important for the activation and attraction of T cell and may influence the course and control of the infection in resistant Balb/c mice.

Standardization of a rapid quadruplex PCR method for the simultaneous detection of bovine, buffalo, Salmonella spp, and Listeria monocytogenes DNA in milk / [Padronização de um método rápido de PCR quadriplex para a detecção simultânea de DNA de bovino, bubalino, Salmonella spp. e Listeria monocytogenes em leite]

The objective of the present study was to Standardize a Polymerase Chain Reaction (PCR) protocol for the authentication of bovine and buffalo milk, and to detect the presence of Salmonella spp. and Listeria monocytogenes. For this, the target DNA was extracted, mixed, and subjected to a PCR assay. Milk samples were defrauded and experimentally contaminated with microorganisms to assess the detection of target DNA at different times of cultivation, bacterial titers, and concentration of genetic material. In addition, the protocol was tested with DNA extracted directly from food, without a pre-enrichment step. The proposed quadruplex PCR showed good accuracy in identifying target DNA sequences. It was possible to simultaneously identify all DNA sequences at the time of inoculation (0h), when the samples were contaminated with 2 CFU/250mL and with 6h of culture when the initial inoculum was 1 CFU/250mL. It was also possible to directly detect DNA sequences from the food when it was inoculated with 3 CFU/mL bacteria. Thus, the proposed methodology showed satisfactory performance, optimization of the analysis time, and a potential for the detection of microorganisms at low titers, which can be used for the detection of fraud and contamination.

O objetivo do presente estudo foi padronizar um protocolo de reação em cadeia da polimerase (PCR) para a autenticação de leite bovino e bubalino e a detecção da presença de Salmonella spp. e Listeria monocytogenes. Para isso, o DNA-alvo foi extraído, misturado e submetido ao ensaio de PCR. Amostras de leite foram fraudadas e contaminadas experimentalmente com os micro-organismos, para se avaliar a detecção do DNA-alvo em diferentes tempos de cultivo, os títulos bacterianos e a concentração de material genético. Além disso, o protocolo foi testado com DNA extraído diretamente do alimento, sem a etapa de pré-enriquecimento. A PCR quadriplex proposta mostrou boa precisão na identificação de sequências de DNA-alvo. Foi possível identificar simultaneamente todas as sequências de DNA no momento da inoculação (0h), quando as amostras estavam contaminadas com 2 UFC/250mL, e com seis horas de cultura, quando o inóculo inicial foi de 1 UFC/250mL. Também foi possível detectar diretamente as sequências de DNA do alimento quando este foi inoculado com 3 UFC/mL de bactérias. Dessa forma, a metodologia proposta apresentou desempenho satisfatório, otimização do tempo de análise e potencial para detecção de micro-organismos em baixos títulos, podendo ser utilizada para detecção de fraude e contaminação.

Hylax bahiensis Bechyné (Chrysomelidae: Eumolpinae): a New Potential Pest of Eucalyptus and Species Used for Atlantic Rainforest Restoration.

Hylax bahiensis Bechyné (Coleoptera: Chrysomelidae), a new pest of forest species, including eucalyptus (hybrid Eucalyptus urophylla x Eucalyptus grandis), Joannesia princeps, Mimosa artemisiana, Croton urucurana, Croton floribundus, and Senna multijuga is recorded. The insect attack in clonal eucalyptus plantations and in forest restoration areas between 2010 and 2013 in the states of Espírito Santo, Bahia and Minas Gerais, Brasil, was observed for the first time. The outbreaks generally occurred from September to March. This new potential pest can affect the growth, productivity, and quality of the trees. We recommended monitoring this leaf-eating beetle especially during the critical period of its occurrence.

Prebiotics and butyric acid can replace colistin as a growth promoter for nursery piglets / [Prebióticos e ácido butírico podem substituir a colistina como promotor de crescimento para leitões em fase de creche]

This study aimed to assess different prebiotic concentrations and principles, in addition to calcium butyrate, aiming to replace colistin as a growth promoter. The sample consisted of 120 piglets weaned at 22 days old with mean initial weight of 5.475 ± 0.719kg. The animals were assigned to random blocks in six treatments corresponding to the use of the following dietary additives: T1) colistin (40 ppm); T2) ß-glucan/mannan-oligosaccharides (0.2%); T3) calcium butyrate (0.1%); T4) ß-glucan/mannan-oligosaccharides (0.1%) + fructooligosaccharides (0.01%) + galactooligosaccharides (0.09%); T5) ß-glucan/mannan-oligosaccharides (0.1%) + fructooligosaccharides (0.03%) + galactooligosaccharides (0.07%); and T6) ß-glucan/mannan-oligosaccharides (0.1%) + fructooligosaccharides (0.05%) + galactooligosaccharides (0.05%). The results showed no difference among treatments for the performance parameters in any of the phases evaluated. For diarrhea incidence and intensity, the results indicated that the treatments with alternative additives had similar effects as the group treated with colistin. A significant difference was found for the profile of propionic acid (0.23% colistin and 0.32%, 0.36%, 0.37% additives) and total fatty acids (0.67% colistin and 0.97% additives) values in the caecum. The supplementation with different compositions and concentrations of prebiotics and butyric acid may viably replace colistin in controlling diarrhea and modulating volatile fatty acid production in the caecum.(AU)

O objetivo deste trabalho foi avaliar as diferentes concentrações e princípios de prebióticos e do butirato de sódio, visando substituir a colistina como promotor de crescimento. Foram utilizados 120 leitões, desmamados aos 22 dias de idade, com peso médio inicial de 5,475 ± 0,719kg. Os animais foram distribuídos em blocos ao acaso, em seis tratamentos, que corresponderam ao uso dos seguintes aditivos dietéticos: T1) colistina (40ppm); T2) ß-glucanos/mananoligossacarídeos (0,2%); T3) butirato de cálcio (0,1%); T4) ß-glucanos/mananoligossacarídeos (0,1%) + frutoligossacarídeos (0,01%) + galactoligossacarídeos (0,09%); T5) ß-glucanos/mananoligossacarídeos (0,1%) + frutoligossacarídeos (0,03%) + galactoligossacarídeos (0,07%); e T6) ß-glucanos/mananoligossacarídeos (0,1%) + frutoligossacarídeos (0,05%) + galactoligossacarídeos (0,05%). Os resultados mostraram que não houve diferença entre os tratamentos para nenhum dos parâmetros de desempenho em nenhuma das fases avaliadas. Para a incidência e a intensidade de diarreia, os resultados apontam que os tratamentos com os aditivos alternativos apresentaram efeitos semelhantes aos do grupo tratado com colistina. Foi encontrada diferença significativa para perfil dos ácidos graxos propiônicos (0,23% colistina e 0,32%, 0,36%, 0,37% aditivos) e ácidos totais (0,67% colistina e 0,97% aditivos) no ceco. A suplementação com diferentes composições e concentrações de prebióticos e do ácido butírico pode substituir a colistina de forma viável no controle da diarreia e na modulação da produção volátil de ácidos graxos no ceco.(AU)

Correlation of pathologic features with clinical outcome in pediatric adrenocortical neoplasia. A study of a Brazilian population. Brazilian Group for Treatment of Childhood Adrenocortical Tumors.

Although the relationship between pathologic features and clinical outcome is well established in adult adrenocortical neoplasms, the prognostic value of these features in pediatric adrenocortical neoplasms (PACN) is unclear. In a series of PACNs from 54 Brazilian children, the authors retrospectively investigated the prognostic value of histologic classification, ploidy, proliferative index, and size (as tumor weight or greater diameter). Histologic classification was most predictive of clinical behavior: there were no failures in 11 adenomas, 5 failures in 27 low-grade carcinomas, and 9 failures in 16 high-grade carcinomas (P = .0003). Tumor weight was predictive of failure in tumors weighing > or = 100 versus < 100 g (P = .04), and a trend was found toward failure among tumors measuring > or = 5 cm, as opposed to those < 5 cm (P = .07). Proliferative index was marginally related to failure (P = .05 at < 11% vs. > or = 11% and .07 at < 10% vs. > or = 10%), and ploidy was not significantly predictive of outcome (P = .25). Histologic type and tumor weight were the most reliable predictors of outcome in PACN.

Stability-indicating colorimetric assay for indicine N-oxide using TLC.

Aliquots of aqueous solutions in which indicine N-oxide may be degraded were mixed with 0.5 M formic acid (1:3) to adjust the pH to approximately 2-4 to quench the reaction and to ensure adequate TLC resolution. Silica-coated aluminum sheets were used to isolate indicine N-oxide by cutting the appropriate region from the chromatogram. By a modification of a known procedure, the silica gel then was treated with an acetic anhydride-diglyme mixture, and the mixture was heated to convert the drug to a pyrrole, which was then coupled with 4-dimethylaminobenzaldehyde to produce a color. The absorbance of the resulting solution was determined at 566 nm, and the apparent molar absorptivity, epsilon, based on the final indicine N-oxide concentration was 6.13 x 10(4). The recovery was approximately 92%, and the assays were readily reproducible with a coefficient of variation of 4.4%.