Aqueous extract OF Moringa oleifera Lam leaves added to freezing extenders damages goat sperm membranes.

BACKGROUND: Semen cryopreservation is a biotechnology used frequently in animal production; however, there are some obstacles, such as those caused by high levels of reactive oxygen species (ROS). Moringa oleifera (MO) is known as a potent source of antioxidants and might be an important adjuvant. OBJECTIVE: The objective of this study was to determine the effect of different concentrations of MO extract supplementation on goat semen cryopreservation efficiency. MATERIALS AND METHODS: Ejaculates (n=6) from four goat breeders were pooled and diluted in skimmed milk (SM) or Tris-egg yolk (TEY)-based extenders and supplemented with different concentrations of MO extract (0, 1, 2 and 5 mg/mL). After the freeze-thaw cycle, sperm kinetics and viability were assessed. RESULTS: With the SM extender, straightness, wobble and plasma membrane integrity were lower than in the control group (P < 0.05). With the TEY extender, wobble was lower in with 5 mg/mL MO extract than in the control group (P < 0.05). As regards sperm ultrastructure, evaluated by SEM, the MO extract, regardless of the diluent used, damaged the membrane of sperm cells in a dose-dependent manner. CONCLUSION: The addition of aqueous extract of MO leaves in both diluents at all concentrations tested affects the parameters of sperm progressivity and damages the plasma membrane in a dose-dependent manner. DOI: 10.54680/fr23310110712.

Clinical study on the efficacy of LED phototherapy for pain control in an orthodontic procedure.

Pain is an unpleasant and emotional subjective sensory experience that occurs during orthodontic procedures. Currently, LED phototherapy is an alternative to the use of laser light as analgesic agent due to similarity of response and lower cost. This case-control, quantitative, qualitative, and longitudinal study aimed to investigate the effect of IR LED phototherapy (λ846 ± 20 nm) in pain during the process of tooth separation during orthodontic treatment. After approval by the Institution Ethics Committee, 40 patients (30 female/10 male, 20-30 years old, average age 24.5 ± 2.6 years old) fulfilling the inclusion criteria entered the study and received a set of four visual analog scales (VAS) for scoring pain immediately, 48 h, 72 h, and 7 days after the insertion of the separating elastics. The patients were randomly distributed into two groups (experimental and control). The patients of experimental group received LED phototherapy (180 mW, 22 s, 4 J, 8 J/cm2, 0.36 W/cm2, spot of 0.5 cm2, spot diameter 0.8 cm) at the same times in which VAS was performed, and control patients were not irradiated. It was found that, in both groups, there was an increase in pain 48 h after insertion of the elastic tooth separator, decreasing 72 h after its installation and reached the lowest level of pain after 7 days. Comparison between groups showed that pain level in the LED group was always statistically significantly lower (p < 0.05), except for the time of installation (T1). The use of LED light was effective in significantly reducing the level of pain after insertion of the elastic tooth separators when compared to the control group.

SETD5 gene variant associated with mild intellectual disability - a case report.

The recent advent of exome sequencing has allowed for the identification of pathogenic gene variants responsible for a variety of diseases that were previously clinically diagnosed, with no underlying molecular etiology. Among these conditions, intellectual disability is a prevalent heterogeneous condition, presenting itself in a large spectrum of intensity, in some cases associated with congenital malformations, behavioral and various other intellectual development alterations. Here we report on a 36-year-old male patient, with a mild intellectual disability that remained undiagnosed at the molecular level for all his life. Using Nextera Exome Sequencing, a Chr3:9.517.294 A>AC (c.3848_3849insC) SETD5 gene insertion was found. This rare variant was classified as likely pathogenic due to its frameshift nature in the gene, in which loss-of-function mutations have been previously reported to cause intellectual disability, as well as a 3p25.3 microdeletion phenotype. It is possible that this variant shows partial activity, due to its gene localization, which would explain the patient's mild phenotype when compared with other reports.

Chemotherapeutical effects of the herbal medicine Uncaria tomentosa (Willd.) DC.

The use of medicinal plants dates back to the beginning of humanity, and today their application as complementary therapy has been widely disseminated as an alternative to conventional therapy. The medicinal plant named Uncaria tomentosa (Willd.) DC. (known as cat's claw) is a common woody vine of the Amazon forest that has traditionally been used in the treatment of arthritis because of its anti-inflammatory properties. This study aimed to evaluate the cytotoxic, mutagenic, and antimutagenic potentials of this medicinal plant. The biological activities of U. tomentosa were determined on bone marrow cells of Wistar rats that were treated in vivo. For the cytotoxic and mutagenic analyses, aqueous plant extract solutions were administered by gavage (1, 2, or 3 mg/mL) for 24 h (an acute treatment) or 7 days (a subchronic treatment). For the antimutagenic analyses, aqueous plant extract solutions (1 mg/mL) were administered by gavage before (pretreatment), simultaneous to (simultaneous treatment), or after (post-treatment), the administration of cyclophosphamide (1.5 mg/mL). U. tomentosa did not show any cytotoxic or mutagenic effects in any of the cytological or chromosomal analyses. Besides, the antimutagenic tests showed that the plant extracts displayed antimutagenic activities, which significantly reduced the percentages of the chromosomal aberrations that were induced by cyclophosphamide at 53.91, 58.60, and 57.03%, respectively, for the simultaneous treatment, pretreatment, and post-treatment. The results suggested a safe use of this herbal medicine that is available free of charge from the Brazilian Public Health System for the treatment of arthritis. This medicinal plant can also effectively contribute to improving the quality of life and the recovery of people undergoing chemotherapeutical treatments.

Vulvar and gastric involvement in plasmablastic lymphoma.

Plasmablastic lymphoma is highly malignant and invasive. It is most commonly found in gastrointestinal tract and is strongly asso- ciated with human immunodeficiency virus (HIV). The authors report a case of a 37-year-old HIV-positive African American woman with a vulvar mass diagnosed as plasmablastic lymphoma. Involvement of the upper gastrointestinal tract had been detected by endoscopic biopsy, suggesting that any patient diagnosed with plasmablastic lymphoma in unusual locations should be investigated for primary lesions in the gastrointestinal tract.

Copper and zinc affect the activity of plasma membrane H+-ATPase and thiol content in aquatic fungi.

Aquatic hyphomycetes are the major microbial decomposers of plant litter in streams. We selected three aquatic hyphomycete species with different abilities to tolerate, adsorb and accumulate copper and zinc, and we investigated the effects of these metals on H+-ATPase activity as well as on the levels of thiol (SH)-containing compounds. Before metal exposure, the species isolated from a metal-polluted stream (Heliscus submersus and Flagellospora curta) had higher levels of thiol compounds than the species isolated from a clean stream (Varicosporium elodeae). However, V. elodeae rapidly increased the levels of thiols after metal exposure, emphasizing the importance of these compounds in fungal survival under metal stress. The highest amounts of metals adsorbed to fungal mycelia were found in the most tolerant species to each metal, i.e. in H. submersus exposed to copper and in V. elodeae exposed to zinc. Short-term (10 min) exposure to copper completely inhibited the activity of H+-ATPase of H. submersus and V. elodeae, whilst zinc only led to a similar effect on H. submersus. However, at longer exposure times (8 days) the most metal-tolerant species exhibited increased H+-ATPase activities, suggesting that the plasma membrane proton pump may be involved in the acclimation of aquatic hyphomycetes to metals.

Fish composition and species richness in eastern South American coastal lagoons: additional support for the freshwater ecoregions of the world.

The relationships between fish composition, connectivity and morphometry of 103 lagoons in nine freshwater ecoregions (FEOW) between 2·83° S and 37·64° S were evaluated in order to detect possible congruence between the gradient of species richness and similarities of assemblage composition. Most lagoons included in the study were <2 km(2) , with a maximum of 3975 km(2) in surface area. Combined surface area of all lagoons included in the study was 5411 km(2) . Number of species varied locally from one to 76. A multiple regression revealed that latitude, attributes of morphometry and connectivity, and sampling effort explained a large amount of variability in species richness. Lagoon area was a good predictor of species richness except in low latitude ecoregions, where lagoons are typically small-sized and not affected by marine immigrants, and where non-native fish species accounted for a significant portion of species richness. Relationships between species and area in small-sized lagoons (<2 km(2) ) is highly similar to the expected number in each ecoregion, with systems located between 18·27° S and 30·15° S attaining higher levels of species richness. Similarities in species composition within the primary, secondary and peripheral or marine divisions revealed strong continental biogeographic patterns only for species less tolerant or intolerant to salinity. Further support for the FEOW scheme in the eastern border of South America is therefore provided, and now includes ecotonal systems inhabited simultaneously by freshwater and marine species of fishes.

Description of a model to optimise the feeding of amino acids to growing pullets.

1. A nutrition model is described that may be used to optimise the amino acid nutrition of laying-type pullets prior to the onset of lay. It is not a method of optimising lifetime laying performance. 2. The potential growth and composition of the body, feathers, ovary and oviduct are described from hatching to the age at which sexual maturity is attained, from which the daily amino acid and energy requirements for the average individual in the population can be calculated. 3. There are two parts to the approach used, the first being a description of the model itself and the second being a description of how the required information was gathered. A number of assumptions made in developing the model are discussed. 4. The rates of maturing of the body, feather-free body and body protein of the DeKalb pullets used were shown to be constant at 0.017/d and those of feathers at 0.02/d. These are considerably slower than those of the oviduct and ovary (0.139 and 0.084/d respectively). The ovary attained a higher mature weight (78.4 vs. 58.7 g) than the oviduct. 5. The age at which the growth of the reproductive organs is initiated in the model is defined by the user. The daily amount of threonine required to meet the requirements for maintenance and potential growth of the developing tissues and organs is calculated for each day of the growing period from hatching to the age at which the first egg is laid. A method of calculating the required daily concentration of threonine in the feed is described, from which a feeding programme may be derived. 6. For the model to produce an optimum economic feeding programme for a population of laying-type pullets, more information than is available from this study is required. These issues are discussed in the paper.

In vitro evaluation of enamel demineralization after several overlapping CO2 laser applications.

This study aimed to evaluate the effects of repeated CO2 laser applications on the inhibition of enamel demineralization. Sixty-five human dental enamel slabs were randomly assigned to the following groups (n = 13): control (C), one application of the CO2 laser (L1), two applications of the CO2 laser (L2), three applications of the CO2 laser (L3), and four applications of the CO2 laser (L4). Enamel slabs were irradiated by a 10.6-µm CO2 laser operating at 5 J/cm(2). The slabs were subjected to a pH-cycling regimen and then analyzed by FT-Raman spectroscopy, energy-dispersive X-ray fluorescence spectrometry (EDXRF), cross-sectional micro-hardness, and scanning electron microscopy (SEM). Statistical analysis was performed using ANOVA and Tukey tests (p < 0.05). FT-Raman spectroscopy showed a reduced carbonate content for L1, L3, and L4 groups when compared to C (p < 0.05). The EDXRF data showed no statistical differences between the control and irradiated groups for calcium and phosphorus components (p > 0.05). Cross-sectional micro-hardness data showed a statistically significant difference between the control and all irradiated groups (p < 0.05), but no difference was found among the irradiated groups (p > 0.05) up to 30-µm depth. A tendency of lower demineralization occurred in deeper depths for L3 and L4 groups. The SEM results showed that with repeated applications of the CO2 laser, a progressive melting and recrystallization of the enamel surface occurred. Repeated irradiations of dental enamel may enhance the inhibition of enamel demineralization.

Tris-egg yolk-glycerol (TEY) extender developed for freezing dog semen is a good option to cryopreserve bovine epididymal sperm cells.

Cryopreservation of epididymal spermatozoa is often performed after shipping the excised testis-epididymis complexes, under refrigeration, to a specialized laboratory. However, epididymal spermatozoa can be collected immediately after excision of the epididymis and sent extended and refrigerated to a laboratory for cryopreservation. In this experiment, we evaluated the effect of both methods of cold storage bovine epididymal spermatozoa as well as of two different extenders on spermatozoa characteristics after freeze-thawing. For that, spermatozoa collected from the caudae epididymis of 19 bulls were extended and cryopreserved in either AndroMed(®) or a Tris-egg yolk (TEY)-based extender. Cryopreservation of sperm cells was performed immediately after castration (Group A, n = 9) or after cold storage for 24 h diluted in the two extenders and (Group B, n = 9) and also after cold storage for 24 h within the whole epididymis (Group C, n = 10). Sperm subjective progressive motility (light microscopy), plasma membrane integrity (hypoosmotic swelling test) and sperm viability (eosin-nigrosin) were evaluated. In vitro fertilization and culture (IVF) was performed to assess the blastocyst rate. No differences (p > 0.05) were observed on post-thaw sperm parameters between samples from Group A, B and C. TEY extended samples presented a higher (p < 0.01) percentage of progressive motile and live sperm, than those extended in AndroMed(®) . Blastocyst rate after IVF differed only (p < 0.05) between the reference group (IVF performed with frozen semen with known in vitro fertility) and Group A extended in AndroMed(®) . We conclude that when cryopreservation facilities are distant from the collection site, bovine epididymal sperm can be shipped chilled overnight either within the epididymal tail or after dilution without deleterious effect on post-thaw sperm quality. TEY extender was more suitable for cold storage and freezing bovine epididymal sperm, than the commercial extender AndroMed®.

A new species of Centromochlus (Siluriformes, Auchenipteridae, Centromochlinae) from the middle Rio Tocantins basin, Brazil.

A new species of the catfish genus Centromochlus (Auchenipteridae, Centromochlinae) is described. The new species is diagnosed by having numerous dark rounded blotches over the body and fins, dorsal-fin spine with serrations anteriorly and smooth posteriorly, anal fin of mature males with three unbranched and seven branched rays, anterior nuchal plate absent and posterior nuchal plate not extended ventrally. The new species is described from a small stream in the Estação Ecológica Serra Geral de Tocantins, a natural reserve in the centre of the Brazilian Cerrado, close to the watershed between the Rio Tocantins and the Rio São Francisco basins. The new species is possibly the sister taxon to the recently described Centromochlus meridionalis from the upper Rio Tapajós. Those two species share with Centromochlus perugiae, from the upper Amazon and upper Paraguay, derived features associated with the modified anal fin in sexually mature males.

Detection of rotavirus in children with acute gastroenteritis in Porto Velho, Rondonia, Brazil.

Rotavirus group A (RVA) infection was ascertained in 591 fecal samples from children 0 to 6 years old with acute gastroenteritis. The vaccination status was also verified in all 591 subjects, with 302 (51.0 % ± 4.0 %) participants fully vaccinated against rotavirus. Forty-two of the vaccinated children (13.9 % ± 3.9 %) tested positive for RVA infection. Of the 289 unvaccinated children (49.0 % ± 4.0 %), 61 (21.1 % ± 4.7 %) had stools positive for RVA. This study suggests that the proportion of acute diarrhea cases caused by rotavirus was low and that the incidence of rotavirus diarrhea decreased over the study period in both vaccinated and unvaccinated children.

Trastuzumab deruxtecan in human epidermal growth factor receptor 2-positive breast cancer brain metastases: a systematic review and meta-analysis.

BACKGROUND: Trastuzumab deruxtecan (T-DXd) has shown promising results in patients with breast cancer brain metastases (BCBMs). We conducted a systematic review and meta-analysis to evaluate the effectiveness and safety of T-DXd in the human epidermal growth factor receptor 2 (HER2)-positive BCBM population. PATIENTS AND METHODS: We searched PubMed, Embase, and Cochrane Library databases as well as American Society of Clinical Oncology (ASCO), European Society for Medical Oncology (ESMO), and San Antonio Breast Cancer Symposium (SABCS) websites for clinical trials (CTs) and observational studies evaluating T-DXd in patients with HER2-positive BCBM. Heterogeneity was assessed with I2 statistics. Random effects models were used for all statistical analyses, which were carried out using R software (version 4.2.2). RESULTS: Ten studies were included, six CTs (n = 189) and four observational studies (n = 130), with a total of 319 patients. The median progression-free survival was 15 months [95% confidence interval (CI) 13.9-16.1 months]. The objective response rate (ORR) was 61% (95% CI 52% to 70%), and the intracranial (IC)-ORR was 61% (95% CI 54% to 69%). No significant differences in ORR and IC-ORR were observed between CTs and observational studies (P = 0.31 and 0.58, respectively). The clinical benefit rate (CBR) was 80% (95% CI 52% to 94%), and the IC-CBR was 70% (95% CI 54% to 82%). The ORR was 68% (95% CI 57% to 77%) in the subgroup of patients with stable BMs and 60% (95% CI 48%-72%) in patients with active BM, with no significant difference between groups (P = 0.35). CONCLUSIONS: Our systematic review and meta-analysis supports the IC activity of T-DXd in patients with stable BM and active BM. TRIAL REGISTRATION: International Prospective Register of Systematic Reviews (PROSPERO) under the protocol number CRD42023422589.

Incidence of interstitial lung disease and cardiotoxicity with trastuzumab deruxtecan in breast cancer patients: a systematic review and single-arm meta-analysis.

BACKGROUND: Trastuzumab deruxtecan (T-DXd) has been shown to benefit progression-free survival and overall survival in patients with metastatic breast cancer (mBC) after progression on ≥1 human epidermal growth factor receptor 2 (HER2)-targeted therapies. However, interstitial lung disease (ILD) and cardiotoxicity are the most significant toxicities associated with T-DXd. Therefore, we conducted a systematic review and meta-analysis to assess the incidence and severity of these toxicities in mBC patients treated with T-DXd. MATERIALS AND METHODS: We searched PubMed, Cochrane, and Scopus databases, and conferences websites for randomized clinical trials and nonrandomized studies of intervention including HER2-low or HER2-positive mBC patients who received at least one dose of T-DXd. Statistical analysis was carried out using R software. RESULTS: We included 15 studies comprising 1970 patients with a mean follow-up of 13.3 months. Median age ranged from 53 to 59 years, 61.9% were non-Asian, and 67.4% had hormone receptor-positive mBC. In a pooled analysis, the incidence of ILD was 11.7% [222 patients; 95% confidence interval (CI) 9.1% to 15.0%]. Patients receiving T-DXd dose of 6.4 mg/kg developed a significantly higher rate of ILD (22.7%) compared to those receiving a dose of 5.4 mg/kg (9.3%) (P < 0.01). Most cases of ILD (80.2%; 174/217 patients) were mild (grade 1 or 2). Grade 3 or 4 ILD was reported in 29 patients (13.4%), and grade 5 in 14 patients (6.4%). The incidence of decreased left ventricular ejection fraction (LVEF) was 1.95% (95% CI 0.65% to 3.73%), and the QT interval (QTi) prolongation was 7.77% (95% CI 2.74% to 20.11%). Most patients were asymptomatic, but four had LV dysfunction and heart failure (0.26%). CONCLUSIONS: In this meta-analysis of 1970 patients with mBC, treatment with T-DXd was associated with a 11.7% incidence of ILD, 7.7% incidence of prolonged QTi, and 1.9% incidence of reduced LVEF. Early detection and management of T-DXd-related toxicity by a multidisciplinary team may ultimately improve patient outcomes.

Eugenia uniflora: a promising natural alternative against multidrug-resistant bacteria.

This work aimed to evaluate the chemical composition, antioxidant and antimicrobial activities from crude extract and fractions from leaves of Eugenia uniflora Linn. The crude extract was obtained by turbo extraction and their fractions by partitioning. Chromatographic analysis were performed, and the antioxidant capacity was verified by two methods (DPPH• and ABTS•+). The Minimal Inhibitory/Bactericidal Concentration were conducted against twenty-two bacteria, selecting five strains susceptible to extract/fractions and resistant to the antibiotics tested. Ampicillin, azithromycin, ciprofloxacin, and gentamicin were associated with Ethyl Acetate Fraction (EAF) against multidrug-resistant strains in modulatory and checkerboard tests. The chromatographic data showed gallic acid, ellagic acid, and myricitrin in crude extract, with enrichment in the EAF. The electron transfer activity demonstrated in the antioxidant tests is related to the presence of flavonoids. The Gram-positive strains were more susceptible to EAF, and their action spectra were improved by association, comprising Gram-negative bacilli. Synergisms were observed to ciprofloxacin and gentamicin against Pseudomonas aeruginosa colistin-resistant. The results demonstrate that the extract and enriched fraction obtained from the leaves of E. uniflora act as a promising natural alternative against multidrug-resistant bacteria.

Salicylic acid concentrations and forms of application mitigate water stress in sour passion fruit seedlings.

The present study aimed to evaluate concentrations and forms of application of salicylic acid used for water stress mitigation on the gas exchange and growth of yellow passion fruit. The experimental design was arranged in randomized blocks in a 4 × 4 × 2 factorial scheme, with four concentrations of salicylic acid (SA) via foliar application (0.0, 0.7, 1.4, and 2.1 mM), four SA concentrations via fertigation (0.0, 0.7, 1.4, and 2.1 mM), and two irrigation depths estimated based on the actual evapotranspiration - ETr (50 and 100% of ETr), with three replications. Water stress negatively affected the physiology and growth of yellow passion fruit seedlings at 75 days after sowing (DAS). The application of salicylic acid, regardless of the form of application, attenuates the effects of water stress on gas exchange and growth of yellow passion fruit, with the best results obtained when applying a concentration of 1.30 mM via leaf or 0.90 mM via fertirrigation. The combination of foliar application of AS and fertigation contributed to improve photosynthetic and growth parameters under water conditions of 50 and 100% of ETr. The foliar application of AS presents superior responses to the application via fertigation. These results reinforce the hypothesis that the attenuation of water stress by salicylic acid is related to the maintenance of gas exchange, which depends on the concentration and form of application, and studies testing combinations throughout the crop cycle become promising for advances in knowledge from the action of this phytohormone on abiotic stress.

Mineral composition and production of guava under salt stress and salicylic acid.

The limitation in the quality of water sources for irrigation in the semi-arid region of northeastern Brazil is increasingly present, so it is necessary to use water with high concentrations of salts for agricultural production, which makes the use of elicitors essential to mitigate the harmful effects of salinity on plants. Given the above, the objective of this study was to evaluate the effects of foliar application of salicylic acid on the mineral composition and production of guava plants under salt stress conditions in the post-grafting phase. The experiment was carried out under greenhouse conditions, in a randomized block design, in a 2 × 4 factorial scheme, with two levels of electrical conductivity of irrigation water (0.6 and 3.2 dS m-1) and four concentrations of salicylic acid (0, 1.2, 2.4, and 3.6 mM), with three replicates. During the flowering stage of guava, N, P, and K contents accumulated in the leaves according to the following order of concentration: N > K > P. Foliar application of 1.2 mM of salicylic acid increases the leaf contents of N, P, and K in guava plants grown under irrigation with water of 0.6 dS m-1. Water salinity of 3.2 dS m-1 reduces the growth and production components of guava plants.

Morphophysiology and production of bell pepper grown under salt stress and salicylic acid foliar application.

Considering the relevance of bell pepper and the limitations imposed by the deleterious effects of salt stress, especially in semi-arid regions, it is extremely important to establish strategies that can facilitate the use of saline water in vegetable production. In this scenario, this study aimed to evaluate the effect of the frequency of foliar application of salicylic acid on the morphophysiology and production of the bell pepper cv. "All Big" irrigated with saline water. The study was conducted at a greenhouse in Campina Grande - PB. The treatments were distributed in a completely randomized design and set up in a 4 × 4 factorial arrangement with three replications, corresponding to four application frequencies of salicylic acid (F1- No application of salicylic acid, F2 - Weekly application, F3- fortnightly application, and F4- monthly application) and four levels of electrical conductivity of irrigation water - ECw (0.8, 1.6, 2.4 and 3.2 dS m-1). The fortnightly application of salicylic acid at a concentration of 1.0 mM mitigated the effects of salt stress on the morphophysiology and production components of bell pepper cv. All Big cultivated with ECw of up to 2.4 dS m-1, which reinforces the hypothesis that salicylic acid can act as a signaling molecule and reduce the effects of saline stress in bell pepper, enabling the use of brackish water in agricultural activity, mainly in semi-arid regions of northeastern Brazil, which have a shortage of fresh water.

Physiology and production of sugar-apple under water stress and application of proline.

The objective of this study was to evaluate the physiology and production of sugar-apple as a function of irrigation intervals and foliar application of proline under the conditions of Paraíba's semi-arid region. A randomized block design was laid out in a 4 × 2 factorial scheme, with treatments resulting from the combination of four irrigation intervals (1, 4, 8 and 12 days) and two concentrations of proline (0 and 10 mM), with four replicates, and the plot consisted of four usable plants. Increase in irrigation intervals reduced the gas exchange of sugar-apple plants at 298 days after transplanting. Exogenous application of proline at concentration of 10 mM increased contents of chlorophyll a, chlorophyll b, total chlorophyll and carotenoids and fruit fresh mass in plants grown under 12-day irrigation intervals.

Production of mycelial biomass, proteases and protease inhibitors by Ganoderma lucidum under different submerged fermentation conditions.

Ganoderma lucidum is a medicinal mushroom widely recognized as a source of biomolecules with pharmacological properties, however, little is known about the factors that influence the synthesis of bioactive proteins by this fungus when cultivated under submerged fermentation. The objective of this work was to evaluate the production of mycelial biomass and intracellular proteases and protease inhibitors by G. lucidum cultivated under different submerged fermentation conditions. The cultivation was carried out in a medium composed of glucose (10 or 20 g.L-1), soy peptone (2.5 or 5 g.L-1) and yeast extract (5 g.L-1), with incubation under agitation (120 rpm) and non-agitation, totaling 8 experimental conditions. Biomass production was determined from the dry weight, while glucose consumption was estimated by quantification of reducing sugars. The proteins were extracted in NaCl (0.15 M), and the protein extracts were submitted to protein quantification by the Bradford method, total proteolytic activity using azocasein, caseinolytic and fibrinolytic activity in Petri dishes, activity of serine (trypsin and chymotrypsin) and cysteine (papain) protease inhibitors. Cultivation in agitated condition showed higher biomass production with a maximum value of 7 g.L-1, in addition to higher activities of trypsin, chymotrypsin and papain inhibitors, with 154 IU.mg-1, 153 IU.mg-1 e 343 IU.mg-1 of protein, respectively. The non-agitated condition showed a greater potential for obtaining proteins, total proteases, caseinolytic and fibrinolytic enzymes, with maximum values of 433 mg.g-1 of extract, 71 U.mL-1 of extract, 63.62 mm2 and 50.27 mm2, respectively. Thus, a medium composed of soy peptone, yest extract and glucose in a 1:2:4 proportion is recommended, under agitation to produce protease inhibitors, and the non-agitated condition when the target is, mainly caseinolytic and fibrinolytic enzymes.