Sulfated polysaccharides from seaweed as a supplement to Prochilodus brevis sperm freezing medium.

BACKGROUND: Using sulfated polysaccharides (SP) in fish sperm freezing medium promotes cell maintenance. OBJECTIVE: To evaluate the effect of different SP concentrations, extracted from two seaweeds (Gracilaria domingensis and Ulva fasciata), as a supplement to the sperm freezing medium of Prochilodus brevis. MATERIALS AND METHODS: Five semen pools were diluted in a solution composed of 5% glucose, 10 % dimethyl sulfoxide (DMSO) and different SP concentrations (0, 0.5, 1.0, 1.5, 2.0, 2.5 or 3.0 mg/mL). The samples were cryopreserved and, after 7 days, rewarmed and analyzed for morphology, plasma membrane integrity, DNA integrity, mitochondrial activity and sperm kinetics [total motility, progressive motility, curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), linearity (LIN), and wobble (WOB)]. RESULTS: There was no interaction between seaweed and SP concentrations. Similar effects were observed with SP extracted from the two seaweeds, regardless of concentration. When comparing the SP concentrations, regardless of the seaweed, 1.0 mg/mL SP showed better results for VCL and VSL. For VAP and WOB, 1.0 mg/mL SP showed better results, but differed from 3.0 mg/mL. LIN followed the same pattern, but differed from SP at 2.5 and 3.0 mg/mL. For progressive motility, 1.0 mg/mL G. domingensis showed superior results compared to the control. For mitochondrial activity, G. domingensis was superior to U. fasciata, regardless of concentration. The lowest concentrations (0.5 and 1.0 mg/mL) showed the best results, regardless of the seaweed. However, the control was superior to all treatments tested. CONCLUSION: G. domingensis SP at the lowest concentrations might be a potential supplement to the P. brevis freezing medium. doi.org/10.54680/fr22210110412.

The effect of glycosaminoglycans, extracted from the skin of tilapia, in the sperm freezing medium of Colossoma macropomum.

BACKGROUND: Sulfated polysaccharides from the skin of Nile tilapia (Oreochromis niloticus), added to the tambaqui (Colossoma macropomum) semen diluting medium, can be potential antioxidants and promote the maintenance of sperm quality. OBJECTIVE: To evaluate the use of different concentrations of glycosaminoglycans (GAGs) from the skin of Nile tilapia as a supplement in two cryogenic media for tambaqui semen. MATERIALS AND METHODS: Tambaqui males received a single dose of pituitary carp extract. The semen was collected for pool analysis and, later, cryopreserved in liquid nitrogen. The pools were diluted and frozen in a solution containing fish-specific powdered coconut water (ACP-104) and 10% DMSO or 5% Glucose and 10% DMSO and supplemented with different concentrations of GAGs. The controls had no GAGs addition. After 45 days, the samples were thawed by immersion in a water bath and evaluated for membrane and DNA integrity, morphology and sperm kinetics. RESULTS: The parameters of linearity (LIN), straightness (STR) and DNA integrity of sperm frozen in 5% Glucose showed better results than ACP-104. For membrane integrity, concentrations of 0 and 1.0 mg/mL were better than 5 mg/mL. Semen motility in 5% Glucose showed superior results at concentrations lower than 5 mg/mL of GAGs. For VCL and VAP, in ACP-104, 3.0 mg/mL exceeded the other treatments. In 5% Glucose, for VCL, 4.0 mg/mL showed the lowest results compared to concentrations of <3.5 mg/mL and, for VAP, it also differed from 4.5 mg/mL CONCLUSION: Therefore, the skin of Nile tilapia has GAGs, in low concentrations, capable of improving the post-thawed sperm quality of tambaqui, especially in 5% Glucose medium.

Avaliação da técnica de vitrificação de ovários de camundongos da linhagem B6D2F1, pertencentes ao Instituto de Ciência e Tecnologia em Biomodelos - ICTB/Fiocruz-RJ / Evaluation of ovary vitrification technique from B6D2F1 strain belonging to the Institute of Biomodels Science and Technology ICTB / Fiocruz-RJ

Objetivou-se testar a vitrificação de ovários de camundongos do ICTB/Fiocruz. Inicialmente, fez-se coleta e maturação in vitro dos oócitos de ovários a fresco e vitrificados, bem como avaliação de estruturas no cultivo embrionário, pós-fertilização in vitro. Fêmeas B6D2F1 foram eutanasiadas para remoção dos ovários (n=60) e divididas em três grupos: grupo 1 (n=30 animais) - oócito de ovários vitrificados, maturados e fertilizados in vitro (120 fragmentos); grupo 2 (n=15) (controle 1) - oócitos coletados a fresco, maturados e fertilizados in vitro; e grupo 3 (n=15) (controle 2) - oócitos maturados in vivo e fertilizados in vitro. A técnica foi verificada no desenvolvimento embrionário in vitro, que foi avaliado pelo teste de qui-quadrado (BioStat 5.0). Recuperaram-se 123, 224 e 328 oócitos nos G1, G2 e G3, respectivamente. Observaram-se diferenças significativas nas taxas de clivagem às 24 horas (embriões ≥ 2 células) entre G1 (8%) e G2 (32%) (P<0,1) e G1 e G3 (49%) (P<0,05), mas não entre G2 e G3 (P>0,05). Para blastocistos, às 96 horas, os grupos G1, G2 e G3 apresentaram, respectivamente, 6%, 11% e 46%, diferindo significativamente entre eles (P<0,05). A vitrificação de ovários, a maturação oocitária e a fertilização in vitro são alternativas para a produção de embriões de camundongos in vitro.(AU)

This work aimed test ovarian vitrification of hybrid mouse from ICTB/Fiocruz. Protocol collection and oocyte in vitro maturation from fresh and vitrified ovaries was established and embryos were evaluated after fertilization. B6D2F1 females were euthanized for ovarian removal (n= 60) and divided into 3 groups: G1 (n= 30) - ovaries fragmented (n= 120), vitrified, matured and fertilized; G2 (n= 15) - in vitro fertilization of oocytes matured in vitro from fresh ovaries; G3 (n= 15) - ampulla region oocytes in vitro fertilizated. Viability was verified by thawing, oocyte in vitro maturation and fertilization. In vitro embryo development of each group was evaluated by Chi-square test (BioStat 5.0). 123, 224 and 328 oocytes were recovered from G1, G2 and G3, respectively. Significant differences were observed in cleavage rates at 24 hours (embryos with 2 cells or more) between G1 (8%) and G2 (32%) (P< 0.1) and G1 and G3 (49%) (P< 0.05) but not between G2 and G3 (P> 0.05). Blastocysts at 96 hours presented 6%, 11% and 46%, respectively for G1, G2 and G3, differing significantly (P< 0.05). Ovary vitrification, oocyte in vitro maturation and in vitro fertilization were available for the production of in vitro mouse embryos.(AU)

Extraction of protease from Aspergillus tamarii URM 4634 in aqueous two-phase system under continuous and discontinuous process.

Aqueous two-phase systems have been studied for almost a century to separate biomolecules in harmless conditions. Proteases produced by Aspergillus tamarii URM 4634 were extracted in polyethylene glycol (PEG)/phosphate aqueous two-phase system under discontinuous and continuous (perforated rotative discs column) process. On the discontinuous process, it was evaluated the effect of operational conditions (PEG molar mass and its concentration, phosphate concentration and pH) over the partition coefficient, activity yield and purification factor. Protease partitioned to PEG-phase with partition coefficients up to 55.73. The best process parameters were 17.5% of PEG, with molar mass 8000 g·mol-1, 15% of phosphate salt at pH 6, with 113.15% of activity yield and purification factor of 2.62. Under continuous extraction, hold up data showed that 57.1% of the discontinuous phase was available for protein extraction. Further, separation achieved 90.0% of efficiency. The yields surpassed 100% in almost all runs, and the best purification factor was 1.84, with both flows of 2 mL·min-1. Thus, the best operational conditions reached an activity yield of 95.3% and 90.0% of separation efficiency. Hence, aqueous two-phase system PEG/phosphate extraction is an efficient process for separation of proteases produced by Aspergillus tamarii URM 4634, under continuous extraction likewise under discontinuous process.

Identification, characterisation and molecular modelling of two AP endonucleases from base excision repair pathway in sugarcane provide insights on the early evolution of green plants.

Unlike bacteria and mammals, plant DNA repair pathways are not well characterised, especially in monocots. The understanding of these processes in the plant cell is of major importance, since they may be directly involved in plant acclimation and adaptation to stressful environments. Hence, two sugarcane ESTs were identified as homologues of AP endonuclease from the base-excision repair pathway: ScARP1 and ScARP3. In order to understand their probable function and evolutionary origin, structural and phylogenetic studies were performed using bioinformatics approaches. The two predicted proteins present a considerable amino acid sequence similarity, and molecular modelling procedures indicate that both are functional, since the main structural motifs remain conserved. However, inspection of the sort signal regions on the full-length cDNAs indicated that these proteins have a distinct organelle target. Furthermore, variances in their promoter cis-element motifs were also found. Although the mRNA expression pattern was similar, there were significant differences in their expression levels. Taken together, these data raise the hypothesis that the ScARP is an example of a probable gene duplication event that occurred before monocotyledon/dicotyledon segregation, followed by a sub-functionalisation event in the Poaceae, leading to new intracellular targeting and different expression levels.

Resource utilisation and cost of ankylosing spondylitis in Brazil.

OBJECTIVES: The present study describes resource utilisation in patients with ankylosing spondylitis (AS) treated at a tertiary public health facility over a one-year period. It also investigates the direct and indirect costs for society associated with the treatment of AS. METHODS: Ninety AS patients were selected consecutively, and data was analysed retrospectively for one year. Resource utilisation was evaluated through systematic interviews with all patients. Demographic, socioeconomic, and clinical variables were recorded, and questionnaires evaluating quality of life, function, and disease activity were also applied (ASQoL, SF-36, HAQ-S, BASFI, and BASDAI). Estimates of indirect costs were performed using the human-capital approach based on the society perspective. RESULTS: Most of the patients were men (79%), with a mean age and disease duration of 40 and 16 years, respectively. The mean HAQ-S, BASFI and BASDAI scores were 1, 5, and 4, respectively. The mean ASQoL score for the sample was 8, and the mean Short-Form-36 scores were between 48 (body pain and general health) and 81 (emotional role). The average monthly household income for the group was US$ 520. The patients had an average of 6 outpatient visits, 6 physical therapy visits and 30 laboratory exams per patient, per year. The average total cost for society was US$ 4,597 per patient per year, of which 45% were direct costs and 55% were indirect costs. CONCLUSIONS: Ankylosing spondylitis is a disease that represents a considerable burden to Brazil. It is extremely important to carry out studies that assess the costs of chronic diseases, especially in developing nations, in order to determine the best manner of allocating the already scanty resources in such regions.