Immunocytochemical evaluation of aquaporins and cell wall components and their influence on foliar water uptake in Andean Melastomataceae.

Andean ecosystems are characterized by high humidity, mainly from rain and fog events. Because of differences in altitude two Andean ecosystems - sub-Andean forest and Páramo -face different environmental pressures that affect leaf anatomy and cell wall composition and, consequently, species foliar water uptake (FWU) capacity. Here, FWU capacity of eight species in the Melastomataceae was evaluated and found to be related to proportions of cell wall components and aquaporins in the two ecosystems. Cellulose was labelled with Calcofluor white, and aquaporin and pectins were labelled with monoclonal antibodies. There were differences in plant FWU capacity in both ecosystems, with higher FWU capacity in sub-Andean forest species than in Páramo forest species. Cell wall components were positively related to FWU, with increased FWU related to pectin and aquaporin content of the plasma membrane. Differences in water availability in the two analysed environments led to differences in FWU capacity that are associated with leaf anatomical traits and cell wall composition. In these two environments, plants with similar traits are selected to respond to given environmental pressures. Traits that favour FWU in sub-Andean forest species may lead to further advances of these species in this environments.

In silico investigation of the role of miRNAs in a possible developmental origin of prostate cancer in F1 and F2 offspring of mothers exposed to a phthalate mixture.

A previous study using miRNA sequencing revealed that exposure to a mixture of phthalates during pregnancy and lactation dysregulated rno-miR-184 and rno-miR-141-3p in the ventral prostate (VP) of offspring. Here, rno-miR-184 and rno-miR-141-3 expressions were obtained by RT-qPCR in the VP of F1 males as well as in F2 offspring, aiming to establish a relationship with possible oncogenic targets through in silico analyses with multigenerational approach. Additionally, some targets were measured by western blots to highlight a possible relationship between the deregulated miRNAs and some of their targets. VP samples from rats exposed to a mixture of phthalates maternally during pregnancy and lactation (GD10 to PND21-F1) and VP from offspring (F2) were examined. The phthalate mixture at both concentrations (20 µg and 200 mg/kg/day) increased the expression of both miRNAs in the F1 (PND22 and 120) and F2 (descendants of F1-treated males) prostate. Target prediction analysis revealed that both microRNAs are responsible for modulating the expression and synthesis of 40 common targets. A phthalate target association analysis and the HPA database showed an interesting relationship among these possible miRNAs modulated targets with prostate adenocarcinoma and other oncogenic processes. Western blots showed alteration in P63, P53, WNT5, and STAT3 expression, which are targeted by the miRNAs, in the VP of F1/F2 males. The data draw attention to the epigenetic modulation in the prostate of descendants exposed to phthalates and adds to one of the few currently found in the literature to point to microRNAs signature as biomarkers of exposure to plasticizers.

Molecular identification of sharks from the genus Sphyrna (Elasmobranchii: Chondrichthyes) in Maranhão Coast (Brazil).

Sharks of the genus Sphyrna are under intense exploitation globally. In Brazil's northern coast, this genus represents a high proportion of fisheries landings and comprises four species. However, due to difficulty of specific identification when specimens are landed, most of the records are limited to the genus level. Here we analyzed the effectiveness of ITS2 (Internal Transcribed Spacer 2 of rDNA) fragment length protocol (Abercrombie et al., 2005) for identifying hammerhead shark species, comparing with the analysis of COI (Cytochrome oxidase subunit I) and ITS2 sequences. We evaluated samples of muscle tissue acquired in the main fishing ports of Maranhão: Carutapera, Raposa e Tutóia. Sampling was conducted between March 2017 to March 2018 and complemented with material deposited in collection (2015). COI results indicated the occurrence of endangered species which are prohibited to be landed. These include Sphyrna mokarran (67%), S. lewini (15%), S. tudes (3%), and S. tiburo (15%). For the ITS2 marker, we investigated the optimization of the protocol developed by Abercrombie (2005) for to improve the use in this geographical area througout design of a new primers.

Integrated transcriptome and proteome analysis indicates potential biomarkers of prostate cancer in offspring of pregnant rats exposed to a phthalate mixture during gestation and lactation.

As the second leading cause of death for cancer among men worldwide, prostate cancer (PCa) prevention and detection remain a critical challenge. One aspect of PCa research is the identification of common environmental agents that may increase the risk of initiation and progression of PCa. Endocrine disrupting chemicals (EDCs) are strong candidates for risk factors, partially because they alter essential pathways for prostate gland development and oncogenesis. Phthalates correspond to a set of commercially used plasticizers that humans are exposed to ubiquitously. Here, we show that maternal exposure to a phthalate mixture interferes with the expression profile of mRNA and proteins in the ventral prostate of offspring and increases the susceptibility to prostate adenocarcinomas in aged animals. The data highlight Ubxn11, Aldoc, Kif5c, Tubb4a, Tubb3, Tubb2, Rab6b and Rab3b as differentially expressed targets in young and adult offspring descendants (PND22 and PND120). These phthalate-induced targets were enriched for pathways such as: dysregulation in post-translational protein modification (PTPM), cell homeostasis, HSP90 chaperone activity, gap junctions, and kinases. In addition, the Kif5c, Tubb3, Tubb2b and Tubb4a targets were enriched for impairment in cell cycle and GTPase activity. Furthermore, these targets showed strong relationships with 12 transcriptional factors (TF), which regulate the phosphorylation of eight protein kinases. The correlation of TF-kinases is associated with alterations in immune system, RAS/ErbB/VEGF/estrogen/HIF-1 signaling pathways, cellular senescence, cell cycle, autophagy, and apoptosis. Downregulation of KIF5C, TUBB3 and RAB6B targets is associated with poor prognosis in patients diagnosed with adenocarcinoma. Collectively, this integrative investigation establishes the post-transcriptional mechanisms in the prostate that are modulated by maternal exposure to phthalate mixture during gestation and lactation.

Effect of plasma and heat treatments on orange juice quality.

Heat treatment is used in the orange juice industry to neutralize the action of pathogenic microorganisms. However, it can reduce the nutritional value of the juice. Thus, our study assessed the cold plasma treatment as an alternative method against Escherichia coli and Candida albicans in 'Lima' orange juice. Both, plasma and heat treatments, reduced the amount of E. coli in the juice, inactivating 16.72 and 100%, respectively. Plasma did not inactivate C. albicans, but heat treatment inactivated 100%. Plasma and heat treatment increased Hue angle and luminosity (more yellowish juice). Plasma reduced vitamin C, carotenoids, and polyphenols content, while increased flavonoids. Heat treatment reduced the carotenoid content. However, neither heat nor plasma treatment altered the antioxidant activity. The plasma treatment reduced the intensity of color (chroma), the soluble solids content and the acidity ratio, total sugars, and the vitamin C content of juice compared to the heat-treated and control juices. Plasma-treated juice showed increased levels of yellow flavonoids, total phenolics and antioxidant activity until the 12th day of storage.

Prevalence of negative behaviour in the dental setting and association with sociodemographic, oral health-related and psychosocial factors amongst Brazilian preschool children.

PURPOSE: To investigate the prevalence of negative behaviour in preschool children attending dental clinics and its association with sociodemographic, oral health-related and parental psychosocial factors. METHODS: A cross-sectional study was carried out with 145 parents/guardians and their children aged 4-6 years attending paediatric dentistry training programmes in a capital city of Midwest Brazil. Data were obtained from children's dental records, interviews, and questionnaires for parents/guardians. The outcome was negative child behaviour, based on the dentists' use or indication of behavioural control measures during the dental appointments, as registered in the children's dental records. Covariates were sociodemographic, clinical and parent/guardian psychosocial factors religiosity (DUREL index) and Sense of Coherence (SOC-13 scale). Bivariate analyses and Poisson regression with robust variance were performed. RESULTS: The prevalence of negative behaviour was 24.1% (95% CI = 17.9-31.7). In the bivariate analyses, the variables initially selected for the regression models (p < 0.25) were the parent/guardians' number of children and religiosity, and the children's dental pain and caries status in deciduous teeth. After adjustment, the prevalence of negative behaviour was 2.12 higher in children with teeth extracted due to caries. CONCLUSION: The prevalence of negative behaviour was high and associated with the presence of missing teeth due to caries, regardless of sociodemographic, psychosocial, and other oral health factors.

Nociception associated with pain/distress in young children sedated for dental treatment: a clinical study with objective and subjective measures.

PURPOSE: To evaluate the association between children's pain/distress levels and patient characteristics (age, sex, history of dental pain), sedation type (level of sedation, sedative regimen), nociception and pain intensity reported by the parents in sedated children undergoing minimally invasive dental treatment. METHODS: This clinical study evaluated secondary outcomes of an RCT that evaluated the efficacy of dexmedetomidine sedation with or without ketamine. Only children who had nociception levels recorded using the Analgesia Nociception Index (ANI) were included (0-100; 0 = highest nociception). FLACC scale (Face, Legs, Activity, Cry and Consolability) was used to assess children's pain/distress (0-10; 0 = no pain/distress). Parental proxy report of the child's pain intensity during the treatment was performed using the Visual Analogue Scale (VAS; 0-100; 0 = no pain). The association between ANI, children's age and sex, dental pain history, sedation level, sedative regimen, and VAS with the FLACC was verified using the generalised estimation equation analysis (alpha 0.05). RESULTS: Participants were 31 children (51.6% boys) and their parents (90.3% mothers). Median FLACC was 3.3 (25th-75th percentiles 1.7-8.2), ANI 80.9 (74.7-85.8), and VAS 10 (1-23). FLACC associated with ANI (OR 0.96; 95% CI 0.93-0.99; p = 0.02), moderate sedation (0.13; 0.03-0.50; p = 0.003), and VAS (1.05; 1.01-1.10; p = 0.01). CONCLUSION: Pain/distress was generally low and more prominent with minimal (versus moderate) sedation and higher nociception.

Effect of plasma and heat treatments on orange juice quality / Efeito de plasma e tratamento térmico na qualidade do suco de laranja

Heat treatment is used in the orange juice industry to neutralize the action of pathogenic microorganisms. However, it can reduce the nutritional value of the juice. Thus, our study assessed the cold plasma treatment as an alternative method against Escherichia coli and Candida albicans in 'Lima' orange juice. Both, plasma and heat treatments, reduced the amount of E. coli in the juice, inactivating 16.72 and 100%, respectively. Plasma did not inactivate C. albicans, but heat treatment inactivated 100%. Plasma and heat treatment increased Hue angle and luminosity (more yellowish juice). Plasma reduced vitamin C, carotenoids, and polyphenols content, while increased flavonoids. Heat treatment reduced the carotenoid content. However, neither heat nor plasma treatment altered the antioxidant activity. The plasma treatment reduced the intensity of color (chroma), the soluble solids content and the acidity ratio, total sugars, and the vitamin C content of juice compared to the heat-treated and control juices. Plasma-treated juice showed increased levels of yellow flavonoids, total phenolics and antioxidant activity until the 12th day of storage.

O tratamento térmico é utilizado na indústria de suco de laranja para neutralizar a ação de microrganismos patogênicos. No entanto, pode reduzir o valor nutricional do suco. Assim, o estudo avaliou o tratamento com plasma frio como método alternativo contra Escherichia coli e Candida albicans em suco de laranja 'Lima'. Ambos os tratamentos, plasma e calor, reduziram a quantidade de E. coli no suco, inativando 16.72 e 100%, respectivamente. O plasma não inativou C. albicans, mas o tratamento térmico inativou 100%. Plasma e tratamento térmico aumentaram o ângulo Hue e a luminosidade do suco (tom mais amarelo). O plasma reduziu o conteúdo de vitamina C, carotenóides e polifenóis, enquanto aumentou os flavonóides. O tratamento térmico reduziu o teor de carotenóides. No entanto, nem o calor nem o tratamento com plasma alteraram a atividade antioxidante. O tratamento com plasma reduziu a intensidade da cor (croma), o teor de sólidos solúveis e a razão de acidez, açúcares totais e o teor de vitamina C do suco em comparação com os sucos tratados termicamente e controle. O suco tratado com plasma apresentou maiores teores de flavonóides amarelos, fenólicos totais e atividade antioxidante até o 12º dia de armazenamento.

Efficacy of hydrogen peroxide wipes for decontamination of AZD1222 adenovirus COVID-19 vaccine strain on pharmaceutical industry materials.

This study aimed to evaluate the performance of accelerated hydrogen peroxide® wipes (HPW) for decontamination of the chimpanzee adenovirus AZD1222 vaccine strain used in the production of recombinant COVID-19 vaccine in a pharmaceutical industry. Two matrices were tested on stainless-steel (SS) and low-density-polyethylene (LDP) surfaces: formulated recombinant COVID-19 vaccine (FCV) and active pharmaceutical ingredient (API). The samples were spiked, dried and the initial inoculum, possible residue effect (RE) and titre reduction after disinfection with HPW were determined. No RE was observed. The disinfection procedure with HPW resulted in complete decontamination the of AZD1222 adenovirus strain in FCV (≥7·46 and ≥7·49 log10 infectious unit [IFU] ml-1 for SS and LDP carriers respectively) and API (≥8·79 and ≥8·78 log10 IFU ml-1 for SS and LDP carriers respectively). In conclusion, virucidal activity of HPW was satisfactory against the AZD1222 adenovirus strain and can be a good option for disinfection processes of SS and LPD surfaces in pharmaceutical industry facilities during recombinant COVID-19 vaccine production. This procedure is simple and can be also applied on safety unit cabins and sampling bags made of LDP as well.

Atypical chronic copper poisoning in a sheep secondary to copper wire ingestion: case report / Intoxicação crônica por cobre atípica em ovino pelo consumo de fios de cobre: relato de caso

A 14-month-old female Texel sheep that came from a herd made up of 19 animals showed haemoglobinuria, apathy, and anorexia, and died two days after the start of the clinical signals. The sheep remained in a natural grassland, where trailers were repaired, and multiple copper wires were deposited on the pasture. The animal had tachycardia, tachypnoea, pale mucous membranes, groaning pain on abdominal palpation, circling, head pressing, intensely hemolyzed plasma, and intense azotaemia. The necropsy showed focally extensive oedema in the inguinal and medial region of pelvic limbs, kidneys dark brown, and liver diffusely yellow with an evident moderate diffuse lobular pattern. The abomasum had a considerable amount of enameled material of thickness, firm to the cut, with 1-5 mm (copper wires). Histopathological examination showed marked diffuse tubular and glomerular coagulative necrosis in the kidneys, in addition to neutrophils, macrophages, lymphocytes, and plasma cells with moderate multifocal nephritis. The liver showed centrilobular necrosis, moderate hepatocellular edema, multifocal cholestasis, and in the lungs and brain mild to moderate diffuse edema. Copper content in the frozen liver (in natura) reached 1,598 mg/kg. Copper mesh ingestion led to sheep poisoning, which in this case was considered an atypical form of chronic primary copper poisoning.

Um ovino Texel de 14 meses de idade, que fazia parte de um rebanho de 19 animais, apresentou hemoglobinúria, apatia, anorexia e morreu dois dias após o início dos sinais clínicos. Os ovinos permaneciam em campo nativo, onde eram realizados consertos de trailers, e múltiplos fios de cobre ficavam depositados na pastagem. O animal apresentou taquicardia, taquipneia, mucosas pálidas, gemido de dor à palpação abdominal, além de andar em círculo, e pressão da cabeça contra obstáculos, plasma intensamente hemolisado e azotemia intensa. Na necropsia, havia edema na região inguinal e medial de membros pélvicos focalmente extenso, rins enegrecidos, e o fígado estava difusamente amarelado, com padrão lobular evidente difuso moderado. No abomaso, havia grande quantidade de material esmaltado, com 1-5mm de espessura, firme, que rangia ao ser cortado (fios de cobre). No exame histopatológico nos rins, havia necrose tubular e glomerular hemoglobinúrica difusa acentuada, além de nefrite de neutrófilios, macrófagos, linfócitos e plasmócitos multifocal moderada. No fígado, havia necrose centrolobular, tumefação hepatocelular e colestase multifocais moderadas; nos pulmões e no cérebro, edema difuso discreto a moderado. A dosagem de Cu no fígado revelou a presença de 1598mg/kg. A ingestão de malhas de cobre levou à intoxicação do ovino que, nesse caso, foi considerada uma forma atípica de intoxicação primária crônica por cobre.

Case report of Curtobacterium isolated from a catheter tip sample misidentified as Cronobacter.

The Curtobacterium genus is a member of the family Microbacteriaceae, and Curtobacterium species are recognized as plant pathogens. The aim of this study was to investigate a dubious result of species identification for an infection located on a catheter tip of a patient with Covid-19. A strain isolated from a catheter tip sample, identified by VITEK® 2 as Cronobacter spp., was submitted to polyphasic analysis: Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF MS) using VITEK® MS, real-time polymerase chain reaction targeting dnaG gene, and 16S rRNA full gene Sanger sequencing analysis for confirmation. The strain presented negative result using qPCR and could not identified by MALDI-TOF MS. 16S rRNA full gene Sanger sequencing analysis identified the strain as Curtobacterium spp. The Gram-variable characteristic (Gram-negative instead of Gram-positive) of the isolated strain was the responsible for the misidentification by VITEK® 2 and VITEK® MS did not identify the strain. 16S rRNA full gene sequencing analysis identified the strain as Curtobacterium genus, but other complementary techniques are necessary to identify at species level.

Core stabilisation exercises reduce chronic low back pain in Air Force fighter pilots: a randomised controlled trial.

BACKGROUND: There is evidence that core stabilisation, strength and endurance training contribute to reduce low back pain in the general population. However, we are not aware of the effect of these exercises on fighter aviation pilots. Therefore, the present study aims to investigate the effects of an exercise protocol on chronic low back pain in Brazilian Air Force fighter pilots. Changes in neck pain, lumbar disability, range of motion, strength and resistance were also investigated. METHODS: Fourteen participants with chronic low back pain were randomised into two groups: stabilisation exercise group (SEG-n=7), exercise protocol twice a week for 12 weeks) and the regular exercise group (REG-n=7), which performed their own usual exercise routine. The evaluations were carried out before and after the training period. The primary outcome was the intensity of low back pain and the secondary outcomes were cervical pain, functional disability, range of motion, maximum isometric strength and trunk muscle resistance. RESULTS: The SEG had a significant reduction in low back pain compared with the REG (difference of 2.3 points, p=0.04) and a lower rate of cervical pain (difference of 2.5 points, p=0.01) at the end of the protocol. Maintenance of trunk muscle strength was also observed in the SEG over the period, while the REG presented a decrease in flexural strength to the right side (difference between groups: -3.7%, p=0.04). There were no differences in the rates of disability, range of motion or resistance to fatigue between groups. CONCLUSIONS: The stabilisation exercise programme focused on the core muscles was successful in decreasing the fighter pilots' chronic lumbar pain. Pilots are normally active; however, our results indicate that specific trunk training programmes should be offered considering the work needs of these subjects. CLINICAL TRIALS: 'Exercise protocol for pilots with back pain' (registered: 09 October 2018)-NCT03713814 (https://clinicaltrials.gov/ct2/show/NCT03713814).

Longevity of Resin Composite and Amalgam Posterior Restorations: A Systematic Review.

OBJECTIVES: The objective of this systematic review was to compare the longevity of direct amalgam and composite resin restorations, in posterior teeth, through clinical, prospective or retrospective studies, with at least 5 years of follow-up. MATERIALS AND METHODS: Studies published in the last 15 years (from 2006 to 2021) were collected using the PubMed and Medline databases. RESULTS: The search strategy associated with the established inclusion and exclusion criteria resulted in a total of 17 articles. Factors related to failures in the performance of restorations were analyzed together with the clinical performance results of each material over the years of study, according to the methodology of each article. CONCLUSIONS: Regardless of the restorative material, the successful results over more than 5 years are due much more to the correct application of the technique, the operator's skill/knowledge and factors related to the patient, such as the type of tooth, number of faces involved in the restoration and oral hygiene.

Polycystic ovary syndrome: clinical and laboratory variables related to new phenotypes using machine-learning models.

PURPOSE: Polycystic Ovary Syndrome (PCOS) is the most frequent endocrinopathy in women of reproductive age. Machine learning (ML) is the area of artificial intelligence with a focus on predictive computing algorithms. We aimed to define the most relevant clinical and laboratory variables related to PCOS diagnosis, and to stratify patients into different phenotypic groups (clusters) using ML algorithms. METHODS: Variables from a database comparing 72 patients with PCOS and 73 healthy women were included. The BorutaShap method, followed by the Random Forest algorithm, was applied to prediction and clustering of PCOS. RESULTS: Among the 58 variables investigated, the algorithm selected in decreasing order of importance: lipid accumulation product (LAP); abdominal circumference; thrombin activatable fibrinolysis inhibitor (TAFI) levels; body mass index (BMI); C-reactive protein (CRP), high-density lipoprotein cholesterol (HDL-c), follicle-stimulating hormone (FSH) and insulin levels; HOMA-IR value; age; prolactin, 17-OH progesterone and triglycerides levels; and family history of diabetes mellitus in first-degree relative as the variables associated to PCOS diagnosis. The combined use of these variables by the algorithm showed an accuracy of 86% and area under the ROC curve of 97%. Next, PCOS patients were gathered into two clusters in the first, the patients had higher BMI, abdominal circumference, LAP and HOMA-IR index, as well as CRP and insulin levels compared to the other cluster. CONCLUSION: The developed algorithm could be applied to select more important clinical and biochemical variables related to PCOS and to classify into phenotypically different clusters. These results could guide more personalized and effective approaches to the treatment of PCOS.

Altitudinal gradient drives regional and local diversity and composition patterns of epiphyllous bryophytes in ecological refuges.

'Brejos de altitude' is an ecosystem that has been subjected to severe exploitation, leading to an intense reduction in Brazil. Understanding the patterns of bryophyte diversity and composition, particularly of specialized species such as the epiphylls, to anthropic and abiotic variables is crucial for implementing protection measures. We investigated the relationship between composition and diversity of epiphyllous bryophytes and anthropic (edge effects) and abiotic (altitude) variables at local (within each 'Brejo') and regional (set of three 'Brejos') scale. Abiotic and anthropic variables were correlated with alpha and beta diversity (decomposed into species replacement and richness differences) using GLM. Their effects on species composition were evaluated using PERMANOVA. The localities at higher elevations harboured a richer bryoflora and overall beta diversity patterns were associated with altitude and locality, which acted over different scales. Regionally, the contribution of richness differences was limited with increasing altitude. Yet, dissimilarities among localities were associated with richness differences and replacement, denoting the importance of local factors. The composition was explained by local factors interacting with the regional altitudinal gradient. Anthropic activity was significant only when interacting with local factors and altitudinal gradient. Environmental filtering associated with altitude played a more important role in shaping the diversity and composition of epiphyllous bryophytes, at both regional and local scales. Each 'Brejo' acts as an ecological refuge, harbouring part of the whole environmental gradient, and presents a unique floristic composition. Moreover, partitioning beta diversity highlighted the interplay of local and regional forces shaping diversity patterns.

Factors associated with low back pain in air force fighter pilots: a cross-sectional study.

INTRODUCTION: Low back pain in military pilots is a frequent condition which constantly leads to absences from work, decreased concentration and performance during flight, as well as changes in work functions. METHODS: This is a cross-sectional analytical study including 28 fighter pilots who underwent an evaluation comprising muscle strength and fatigue resistance, trunk mobility and application of questionnaires to identify associated clinical factors. RESULTS: It was observed that 68% of the pilots reported low back pain with an average pain intensity of 3.7 at numerical pain scale in the last week. No significant differences were observed regarding the range of motion and trunk muscle strength when pilots with low back pain were compared with asymptomatic pilots. However, lateral right trunk muscle (mean difference=16, 95% CI 0.6 to 33.0]) and lateral left trunk muscle (mean difference=22, 95% CI 1 to 44) fatigue sooner in pilots with low back pain when the two groups were compared (p=0.04 for both). CONCLUSION: There was a high rate of low back pain complaints among fighter pilots. There was also a significant reduction in fatigue resistance of the lateral trunk muscles in symptomatic pilots when compared with asymptomatic pilots. These factors must be considered in the physical training of this population.

Obtainment and characterization of digestive aspartic proteases from the fish Caranx hippos (Linnaeus, 1766) / Obtenção e caracterização de proteases aspárticas digestórias do peixe Caranx hippos (Linnaeus, 1766)

This work aimed to obtain aspartic proteases of industrial and biotechnological interest from the stomach of the crevalle jack fish (Caranx hippos). In order to do so, a crude extract (CE) of the stomach was obtained and subjected to a partial purification by salting-out, which resulted in the enzyme extract (EE) obtainment. EE proteases were characterized physicochemically and by means of zymogram. In addition, the effect of chemical agents on their activity was also assessed. By means of salting-out it was possible to obtain a purification of 1.6 times with a yield of 49.4%. Two acid proteases present in the EE were observed in zymogram. The optimum temperature and thermal stability for EE acidic proteases were 55 ºC and 45 °C, respectively. The optimum pH and pH stability found for these enzymes were pH 1.5 and 7.0, respectively. Total inhibition of EE acid proteolytic activity was observed in the presence of pepstatin A. dithiothreitol (DTT) and Ca2+ did not promote a significant effect on enzyme activity. In the presence of heavy metals, such as Al3+, Cd2+ and Hg2+, EE acidic proteases showed more than 70% of their enzymatic activity. The results show that it is possible to obtain, from the stomach of C. hippos, aspartic proteases with high proteolytic activity and characteristics that demonstrate potential for industrial and biotechnological applications.

Este trabalho objetivou obter proteases aspárticas de interesse industrial e biotecnológico a partir do estômago do peixe xaréu (Caranx hippos). Para isso, foi obtido um extrato bruto do estômago, o qual foi submetido a uma purificação parcial por salting-out onde se obteve o extrato enzimático (EE). As proteases do EE foram caracterizadas físico-quimicamente e através de zimograma. Além disso, o efeito de agentes químicos sobre sua atividade também foi avaliado. Através de salting-out foi possível obter uma purificação de 1,6 vezes com rendimento de 49,4%. Foram observadas duas proteases ácidas presentes no EE através de zimograma. A temperatura ótima e a estabilidade térmica para as proteases ácidas do EE foram de 55 ºC e 45 °C, respectivamente. O pH ótimo e a estabilidade ao pH encontrados para estas enzimas foram o pH 1,5 e 7,0, respectivamente. Observou-se a inibição total da atividade proteolítica ácida do EE na presença de pepstatina A. O ditiotreitol (DTT) e o Ca2+ não promoveram efeito significativo na atividade enzimática. Na presença de metais pesados, como Al3+, Cd2+ e Hg2+, o EE manteve mais de 70% de atividade enzimática do EE. Os resultados mostram que é possível obter, a partir do estômago de C. hippos, proteases aspárticas com alta atividade proteolítica e características que demonstram potencial para aplicações industriais e biotecnológicas.

Carangoides bartholomaei (Cuvier, 1833) stomach: a source of aspartic proteases for industrial and biotechnological applications / Estômago de Carangoides bartholomaei (Cuvier, 1833): uma fonte de proteases aspárticas para aplicações industriais e biotecnológicas

The viscera and other residues from fish processing are commonly discarded by the fishing industry. These byproducts can be a source of digestive enzymes with industrial and biotechnological potential. In this study, we aimed at the extraction, characterization, and application of acidic proteases from the stomach of Carangoides bartholomaei (Cuvier, 1833). A crude extract from the stomachs was obtained and submitted to a partial purification process by salting-out, which obtained a Purified Extract (PE) with a specific proteolytic activity of 54.0 U·mg-1. A purification of 1.9 fold and a yield of 41% were obtained. The PE presents two isoforms of acidic proteases and a maximum proteolytic activity at 45 °C and pH 2.0. The PE acidic proteolytic activity was stable in the pH range of 1.5 to 7.0 and temperature from 25 °C to 50 °C. Purified Extract kept 35% of its proteolytic activity at the presence of NaCl 15% (m/v) but was totally inhibited by pepstatin A. Purified Extract aspartic proteases presented high activity in the presence of heavy metals such as Cd2+, Hg2+, Pb2+, Al3+, and Cu2+. The utilization of PE as an enzymatic addictive in the collagen extraction from Nile tilapia scales has doubled the process yield. The results indicate the potential of these aspartic proteases for industrial and biotechnological applications.

As vísceras e outros resíduos do processamento de peixes são geralmente descartados pela indústria pesqueira. Esses resíduos podem ser uma fonte de enzimas digestivas com potencial industrial e biotecnológico. Neste estudo, objetivamos a extração, caracterização e aplicação de proteases aspárticas do estômago de Carangoides bartholomaei (Cuvier, 1833). Um extrato bruto do estômago foi obtido e submetido a um processo de purificação parcial, que obteve um Extrato Purificado (EP) com uma atividade proteolítica específica de 54,0 U·mg-1. Foi obtida uma purificação de 1,9 vezes e um rendimento de 41%. O EP apresenta duas isoformas de proteases ácidas e atividade proteolítica máxima a 45 °C e pH 2,0. A atividade proteolítica do EP foi estável na faixa de pH de 1,5 a 7,0 e temperatura de 25 °C a 50 °C. O EP manteve 35% de sua atividade proteolítica na presença de NaCl a 15% (m/v), mas foi totalmente inibida pela pepstatina A. As proteases ácidas do EP apresentaram alta atividade na presença de metais pesados como o Cd2+, Hg2+, Pb2+, Al3+ e Cu2+. A utilização de EP como aditivo enzimático na extração de colágeno a partir de escamas de tilápia do Nilo dobrou o rendimento do processo. Os resultados indicam um potencial dessas proteases para aplicações industriais e biotecnológicas.