Extract of Cimicifuga racemosa (L.) Nutt protects ovarian follicle reserve of mice against in vitro deleterious effects of dexamethasone.

The present study aims to investigate if Cimicifuga racemosa (L.) Nutt extract (CIMI) reduces deleterious effects of dexamethasone (DEXA) in ovaries cultured in vitro. Mouse ovaries were collected and cultured in DMEM+ only or supplemented with 5 ng/mL of CIMI, or 4 ng/mL DEXA, or both CIMI and DEXA. The ovaries were cultured at 37.5°C in 5% CO2 for 6 days. Ovarian morphology, follicular ultrastructure, and the levels of mRNA for Bax, Bcl-2, and Caspase-3 were evaluated. The results showed that DEXA reduced the percentage of morphologically normal follicles, while CIMI prevented the deleterious effects caused by DEXA. In addition, DEXA negatively affected the stromal cellular density, while CIMI prevented these adverse effects. Ovaries cultured with DEXA and CIMI showed similar levels of mRNA for Bax, Bcl-2, and Caspase-3 compared to those cultured in control medium, while ovaries cultured with DEXA had increased expression of the above genes. Additionally, the ultrastructure of the ovaries cultured with CIMI was well preserved. Thus, the extract of CIMI was able to prevent the deleterious effects caused by DEXA on cultured mouse ovaries.

Mesh excision secondary to spondylodiscitis after colposacropexy graft rejection: a step by step video.

INTRODUCTION: Spondylodiscitis secondary to colposacropexy is an extremely rare entity. Infection and mesh rejection are the main causes. Removal of the mesh is essential for patient's recovery and it can be a very challenging surgical procedure. CASE: A 72-year-old woman presented with severe low back pain in the context of a recent colposacropexy. Magnetic resonance imaging was performed and spondylodiscitis secondary to prolapse correction surgery with mesh was suspected. In order to ensure an adequate recovery, removal of the mesh was required. CONCLUSIONS: Spondylodiscitis secondary to colposacropexy should be suspected when the patient starts with moderate lumbar pain and is not correctly controlled with first-level analgesia. Infection or mesh rejection should be considered. Mesh rejection should be suspected when the patient does not improve after antibiotics. Complete removal of the mesh is needed in order to ensure the patient's recovery.

One stage procedure for developmental dysplasia of the hip in patients aged eight years or older. Systematic review and meta-analysis.

The objective of the present study is to perform a systematic review with meta-analysis of the literature on treatment of developmental dysplasia of the hip (DDH) in patients over eight years of age to provide better understanding of therapeutic strategies and results. Authors carried out a systematic review and metanalysis of the literature on DDH treated in patients aged eight years of age or older. A literature search was carefully performed from June 2019 to June 2020. The articles had to report one stage reconstructive surgical treatment of DDH for patients eight years or older, presenting clinical and radiographic evaluation according to the Tonnis and Severin, and McKay systems. Meta-analysis was carried out using the software metanalyst to perform the pooled effect size Nine studies met the inclusion criteria. They assessed a total of 234 patients and 266 hips. Female patients accounted for 75.7% (eight unknown) and the follow-up ranged from 1 to 17.4 years. The majority of the procedures included an acetabular surgery (93.9%) while femoral shortening was performed in 78%. Acceptable outcomes were found in between 67% (Mckay system) and 91% (Severin system) of the cases. Femoral varus and derrotation shortening associated with redirectional osteotomy of the acetabulum (for those with closed triradiate cartilage) or acetabular redirection/reshaping were the most prevalent combined procedures; this strategy can lead to 60% of acceptable clinical results and 90% of radiographically acceptable results. Therefore, our findings give credence to the recommendation for the treatment of DDH in patients over eight years old.

Quantitative MRI evaluation of bone marrow in sickle cell disease: relationship with haemolysis and clinical severity.

AIM: To evaluate bone marrow fat fraction using the Dixon technique (FFDix) of magnetic resonance imaging (MRI) as a potential biomarker of haemolysis and clinical severity in the overall assessment and follow-up of sickle cell disease (SCD) patients. MATERIAL AND METHODS: The present study was a cross-sectional study in which healthy individuals and SCD patients (matched for age, sex, and weight) were subjected to MRI of the lumbar spine and pelvis to quantify FFDix in the bone marrow using the Dixon technique. SCD severity was analysed by clinical and laboratory data, and an online calculator. A high degree of haemolysis was defined using the cut-off values haemoglobin (Hb) ≤10 g/dl, lactate dehydrogenase (LDH) ≥325 U/l, reticulocytes ≥3% and total bilirubin (TB) ≥1.2 mg/dl. Pearson's correlation, receiver operating characteristic (ROC) curve and binary logistic regression analysis were performed. RESULTS: Forty-eight SCD patients (26 homozygous: HbSS and 22 compound heterozygous: HbSC) and 48 healthy individuals participated in the study. FFDix was lower in SCD patients than in the control group, showing even lower values in the HbSS subtype and patients with a higher degree of haemolysis. HbSC patients with a higher degree of haemolysis using hydroxyurea (medium dosage 9.8 mg/kg/day) had lower FFDix. ROC curves and odds ratios for detecting patients with a higher degree of haemolysis at the different FFDix measurement sites demonstrated excellent performance: iliac bones (cut-off ≤16.75%, AUC = 0.824, p<0.001), femoral heads (cut-off ≤46.7%, AUC = 0.775, p=0.001), lumbar vertebrae (cut-off ≤7.8%, AUC = 0.755, p=0.002). CONCLUSION: Decreased FFDix is indicative of higher degree of haemolysis and SCD severity with great potential as a non-invasive biomarker contributing to the overall assessment and follow-up of SCD patients.

Extract of Cimicifuga racemosa (L.) Nutt protects ovarian follicle reserve of mice against in vitro deleterious effects of dexamethasone

The present study aims to investigate if Cimicifuga racemosa (L.) Nutt extract (CIMI) reduces deleterious effects of dexamethasone (DEXA) in ovaries cultured in vitro. Mouse ovaries were collected and cultured in DMEM+ only or supplemented with 5 ng/mL of CIMI, or 4 ng/mL DEXA, or both CIMI and DEXA. The ovaries were cultured at 37.5°C in 5% CO2 for 6 days. Ovarian morphology, follicular ultrastructure, and the levels of mRNA for Bax, Bcl-2, and Caspase-3 were evaluated. The results showed that DEXA reduced the percentage of morphologically normal follicles, while CIMI prevented the deleterious effects caused by DEXA. In addition, DEXA negatively affected the stromal cellular density, while CIMI prevented these adverse effects. Ovaries cultured with DEXA and CIMI showed similar levels of mRNA for Bax, Bcl-2, and Caspase-3 compared to those cultured in control medium, while ovaries cultured with DEXA had increased expression of the above genes. Additionally, the ultrastructure of the ovaries cultured with CIMI was well preserved. Thus, the extract of CIMI was able to prevent the deleterious effects caused by DEXA on cultured mouse ovaries.

Chemical composition and phytotoxicity of essential oils of Croton doctoris S. Moore (Euphorbiaceae) / Composição química e fitotoxicidade dos óleos essenciais de Croton doctoris S. Moore (Euphorbiaceae)

Essential oils from the stems and leaves of Croton doctoris were analyzed by gas chromatography and mass spectrometry, resulting in 22 identified compounds. The effects of these essential oils on the germination, root and shoot growth, total chlorophyll content, potential root respiration, peroxidase activity, catalase, superoxide dismutase, and mitotic index in lettuce and onion were determined. Antioxidant, antimicrobial, and cytotoxic activity were also investigated. The results revealed that the stem oil consisted of 15 compounds, of which caryophyllene oxide (24.5%) and E-caryophyllene (13.3%) were the major constituents. The leaf oil contained E-caryophyllene (39.6%) and α-humulene (13.2%) as major compounds. The oils inhibited the germination and growth of lettuce and onion seedlings and reduced chlorophyll content, root respiration, and cell division. They also caused oxidative stress, indicated by the increased activity of the evaluated antioxidant enzymes. These abnormal physiological processes contributed to the inhibition of plant growth. The most pronounced phytotoxic effects were observed in the stem oil. The cytotoxicity tests indicated that leaf oil was more active than stem oil, resulting from the presence of biologically active sesquiterpenes that inhibit the growth of cancer cells.

Os óleos essenciais do caule e da folha de Croton doctoris foram analisados por cromatografia gasosa (GC) e espectrometria de massa (GC-MS) resultando em 22 compostos identificados. Os efeitos dos óleos essenciais na germinação, crescimento de raízes e parte aérea, teor total de clorofila, respiração radicular, atividade de peroxidase, catalase e superóxido de dimetase e índice mitótico foram determinados em alface e cebola. Atividade antioxidante, antimicrobiana e citotóxica também foram investigadas. Os resultados revelaram que o óleo do caule é constituído por 15 compostos, dos quais os principais são o óxido de cariofileno (24,5%) e E-cariofileno (13,3%). O óleo foliar apresentou E-cariofileno (39,6%) seguido de α-humuleno (13,2%) como compostos majoritários. Os óleos inibiram a germinação e o crescimento das plântulas de alface e cebola e reduziram o conteúdo de clorofila, a respiração radicular e a divisão celular. Eles também causaram estresse oxidativo, indicado pelo aumento da atividade das enzimas antioxidantes avaliadas. Esses processos fisiológicos anormais contribuem para a inibição do crescimento das plantas. Os efeitos fitotóxicos mais pronunciados foram observados no óleo do caule. Nos testes de citotoxicidade observou-se que o óleo das folhas foi mais ativo, resultante da presença de sesquiterpenos biologicamente ativos que atuam inibindo o crescimento das células cancerígenas.

Chemical composition and phytotoxicity of essential oils of Croton doctoris S. Moore (Euphorbiaceae) / Composição química e fitotoxicidade dos óleos essenciais de Croton doctoris S. Moore (Euphorbiaceae)

Essential oils from the stems and leaves of Croton doctoris were analyzed by gas chromatography and mass spectrometry, resulting in 22 identified compounds. The effects of these essential oils on the germination, root and shoot growth, total chlorophyll content, potential root respiration, peroxidase activity, catalase, superoxide dismutase, and mitotic index in lettuce and onion were determined. Antioxidant, antimicrobial, and cytotoxic activity were also investigated. The results revealed that the stem oil consisted of 15 compounds, of which caryophyllene oxide (24.5%) and E-caryophyllene (13.3%) were the major constituents. The leaf oil contained E-caryophyllene (39.6%) and α-humulene (13.2%) as major compounds. The oils inhibited the germination and growth of lettuce and onion seedlings and reduced chlorophyll content, root respiration, and cell division. They also caused oxidative stress, indicated by the increased activity of the evaluated antioxidant enzymes. These abnormal physiological processes contributed to the inhibition of plant growth. The most pronounced phytotoxic effects were observed in the stem oil. The cytotoxicity tests indicated that leaf oil was more active than stem oil, resulting from the presence of biologically active sesquiterpenes that inhibit the growth of cancer cells.

Os óleos essenciais do caule e da folha de Croton doctoris foram analisados por cromatografia gasosa (GC) e espectrometria de massa (GC-MS) resultando em 22 compostos identificados. Os efeitos dos óleos essenciais na germinação, crescimento de raízes e parte aérea, teor total de clorofila, respiração radicular, atividade de peroxidase, catalase e superóxido de dimetase e índice mitótico foram determinados em alface e cebola. Atividade antioxidante, antimicrobiana e citotóxica também foram investigadas. Os resultados revelaram que o óleo do caule é constituído por 15 compostos, dos quais os principais são o óxido de cariofileno (24,5%) e E-cariofileno (13,3%). O óleo foliar apresentou E-cariofileno (39,6%) seguido de α-humuleno (13,2%) como compostos majoritários. Os óleos inibiram a germinação e o crescimento das plântulas de alface e cebola e reduziram o conteúdo de clorofila, a respiração radicular e a divisão celular. Eles também causaram estresse oxidativo, indicado pelo aumento da atividade das enzimas antioxidantes avaliadas. Esses processos fisiológicos anormais contribuem para a inibição do crescimento das plantas. Os efeitos fitotóxicos mais pronunciados foram observados no óleo do caule. Nos testes de citotoxicidade observou-se que o óleo das folhas foi mais ativo, resultante da presença de sesquiterpenos biologicamente ativos que atuam inibindo o crescimento das células cancerígenas.

Chemical composition and phytotoxicity of essential oils of Croton doctoris S. Moore (Euphorbiaceae)

Abstract Essential oils from the stems and leaves of Croton doctoris were analyzed by gas chromatography and mass spectrometry, resulting in 22 identified compounds. The effects of these essential oils on the germination, root and shoot growth, total chlorophyll content, potential root respiration, peroxidase activity, catalase, superoxide dismutase, and mitotic index in lettuce and onion were determined. Antioxidant, antimicrobial, and cytotoxic activity were also investigated. The results revealed that the stem oil consisted of 15 compounds, of which caryophyllene oxide (24.5%) and E-caryophyllene (13.3%) were the major constituents. The leaf oil contained E-caryophyllene (39.6%) and -humulene (13.2%) as major compounds. The oils inhibited the germination and growth of lettuce and onion seedlings and reduced chlorophyll content, root respiration, and cell division. They also caused oxidative stress, indicated by the increased activity of the evaluated antioxidant enzymes. These abnormal physiological processes contributed to the inhibition of plant growth. The most pronounced phytotoxic effects were observed in the stem oil. The cytotoxicity tests indicated that leaf oil was more active than stem oil, resulting from the presence of biologically active sesquiterpenes that inhibit the growth of cancer cells.

Resumo Os óleos essenciais do caule e da folha de Croton doctoris foram analisados por cromatografia gasosa (GC) e espectrometria de massa (GC-MS) resultando em 22 compostos identificados. Os efeitos dos óleos essenciais na germinação, crescimento de raízes e parte aérea, teor total de clorofila, respiração radicular, atividade de peroxidase, catalase e superóxido de dimetase e índice mitótico foram determinados em alface e cebola. Atividade antioxidante, antimicrobiana e citotóxica também foram investigadas. Os resultados revelaram que o óleo do caule é constituído por 15 compostos, dos quais os principais são o óxido de cariofileno (24,5%) e E-cariofileno (13,3%). O óleo foliar apresentou E-cariofileno (39,6%) seguido de -humuleno (13,2%) como compostos majoritários. Os óleos inibiram a germinação e o crescimento das plântulas de alface e cebola e reduziram o conteúdo de clorofila, a respiração radicular e a divisão celular. Eles também causaram estresse oxidativo, indicado pelo aumento da atividade das enzimas antioxidantes avaliadas. Esses processos fisiológicos anormais contribuem para a inibição do crescimento das plantas. Os efeitos fitotóxicos mais pronunciados foram observados no óleo do caule. Nos testes de citotoxicidade observou-se que o óleo das folhas foi mais ativo, resultante da presença de sesquiterpenos biologicamente ativos que atuam inibindo o crescimento das células cancerígenas.

Rutin promotes activation and reduces apoptosis of primordial follicles by regulating Akt phosphorylation after in vitro culture of ovine ovarian tissue.

The aims of this study were to analyze the effects of different concentrations of rutin on primordial follicle survival and development after in vitro culture of sheep ovarian tissue, and to verify the possible involvement of the phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt) pathway in the rutin actions. Ovarian fragments were fixed for histological analysis (fresh control) or cultured in α-minimum essential medium alone (α-MEM+: control medium) or in α-MEM+supplemented with different concentrations of rutin (0.1; 1 or 10 µg/mL) for 7 days. Inhibition of the PI3K activity was performed in fragments cultured with 50 µM LY294002. Thereafter, immunohistochemistry was performed to evaluate the expression of cleaved caspase-3 (apoptosis) and Akt phosphorylation (p-Akt). The results showed that 1 µg/mL rutin has a greater percentage of normal follicles (P < 0.05) than those of α-MEM+ and other rutin treatments. In addition, 1 µg/mL rutin maintained the follicular apoptosis similar (P > 0.05) to that of the fresh control and lower than α-MEM+ and 10 µg/mL rutin. All rutin concentrations increased (P < 0.05) follicular activation compared to fresh control and α-MEM+. Furthermore, follicular and oocyte diameters increased (P < 0.05) only after culture with 1 µg/mL rutin. After PI3K inhibition, there was a reduction (P < 0.05) of rutin follicular effects. In conclusion, rutin at 1 µg/mL reduces apoptosis, promotes activation and growth of sheep primordial follicles through the modulation of the PI3K/Akt signaling pathway after in vitro culture of ovine ovarian tissue.

Occupational infection to Brucella abortus B19 vaccine despite antimicrobial prophylaxis / Infecção ocupacional pela cepa vacinal de Brucella abortus B19 apesar de profilaxia antimicrobiana

ABSTRACT: The authors report the case of a veterinarian who acquired brucellosis infection by accidental exposure to Brucella abortus vaccine (BRUCEL-VET B19) while performing animal vaccination. Antibiotic prophylaxis with doxycycline and rifampin for six weeks was indicated, but rifampin was discontinued after 10 days due to gastrointestinal in-tolerance. Despite prophylaxis, the patient seroconverted after 30 days, but was asymptomatic and did not require additional antibiotic therapy. Post-exposure prophylaxis of Brucella is not free from side effects and asymptomatic seroconversion can occur despite prophylaxis. (AU)

RESUMO: Os autores relatam o caso de um veterinário que adquiriu infecção por brucelose por exposição acidental à vacina Brucellaabortus (BRUCEL-VET B19) durante a vacinação animal. A profilaxia antibiótica com doxiciclina e rifampici-na por seis semanas foi indicada, mas a rifampicina foi descontinuada após 10 dias devido à intolerância gastroin-testinal. A profilaxia pós-exposição de Brucella não está isenta de efeitos colaterais e a soroconversãoassintomática pode ocorrer apesar da profilaxia. (AU)

Chemical composition and phytotoxicity of essential oils of Croton doctoris S. Moore (Euphorbiaceae).

Essential oils from the stems and leaves of Croton doctoris were analyzed by gas chromatography and mass spectrometry, resulting in 22 identified compounds. The effects of these essential oils on the germination, root and shoot growth, total chlorophyll content, potential root respiration, peroxidase activity, catalase, superoxide dismutase, and mitotic index in lettuce and onion were determined. Antioxidant, antimicrobial, and cytotoxic activity were also investigated. The results revealed that the stem oil consisted of 15 compounds, of which caryophyllene oxide (24.5%) and E-caryophyllene (13.3%) were the major constituents. The leaf oil contained E-caryophyllene (39.6%) and α-humulene (13.2%) as major compounds. The oils inhibited the germination and growth of lettuce and onion seedlings and reduced chlorophyll content, root respiration, and cell division. They also caused oxidative stress, indicated by the increased activity of the evaluated antioxidant enzymes. These abnormal physiological processes contributed to the inhibition of plant growth. The most pronounced phytotoxic effects were observed in the stem oil. The cytotoxicity tests indicated that leaf oil was more active than stem oil, resulting from the presence of biologically active sesquiterpenes that inhibit the growth of cancer cells.

Leptin decreases apoptosis and promotes the activation of primordial follicles through the phosphatidylinositol-3-kinase/protein kinase B pathway in cultured ovine ovarian tissue.

This study evaluated the effects of leptin on primordial follicle survival and activation after in vitro culture of ovine ovarian tissue and if leptin acts through the phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt) pathway. Ovarian fragments were fixed for histology (fresh control) or cultured for 7 days in control medium (α-MEM+) alone or supplemented with leptin (1, 5, 10, 25 or 50 ng/ml). Follicle morphology, activation and apoptosis were analyzed. Next, the fragments were cultured in the medium that showed the best results in the absence or the presence of the PI3K inhibitor (LY294002), and immunohistostaining of p-Akt protein was assessed. After culture, the percentage of normal follicles decreased (P < 0.05) in all treatments compared with the fresh control. Moreover, control medium and 1 ng/ml leptin had similar (P > 0.05) percentages of normal follicles, which were significantly higher than those in other treatments. However, culture with 1 ng/ml leptin maintained apoptosis similarly (P > 0.05) to that of the fresh control and lower (P < 0.05) than that in α-MEM+. Leptin did not influence follicle activation (P > 0.05) compared with the control medium (α-MEM+). Culture in 1 ng/ml leptin with LY294002 decreased the normal follicles and increased apoptosis, inhibited follicle activation (P < 0.05), and reduced p-Akt immunostaining, compared with the medium containing 1 ng/ml leptin without PI3K inhibitor. In conclusion, leptin at 1 ng/ml reduces apoptosis and promotes the activation of primordial follicles compared with the fresh control after in vitro culture of ovine ovarian tissue possibly through the PI3K/Akt pathway.

Involvement of Phosphorylated Akt and FOXO3a in the Effects of Growth and Differentiation Factor-9 (GDF-9) on Inhibition of Follicular Apoptosis and Induction of Granulosa Cell Proliferation After In Vitro Culture of Sheep Ovarian Tissue.

This study aimed to evaluate the effects of growth and differentiation factor-9 (GDF-9) on the morphology, activation, apoptosis, and granulosa cell proliferation of ovine preantral follicles cultured within ovarian tissue slices and to verify whether GDF-9 could influence follicular activation through the phosphatidylinositol 3-kinase/protein kinase B/forkhead box O3a (PI3K/Akt/FOXO3a) pathway. Ovine ovarian fragments were cultured in α-MEM+ or α-MEM+ with GDF-9 (1, 50, 100, 200, or 400 ng/ml) for 7 days. Apoptosis and cell proliferation were analyzed. Next, the activation of the PI3K was inhibited with LY294002, and immunostaining for p-Akt and p-FOXO3a proteins was assessed. The concentration of 50 ng/ml GDF-9 had (P < 0.05) more morphologically normal follicles compared to all treatments, except 1 ng/ml GDF-9. Moreover, 50 ng/ml GDF-9 increased primordial follicle activation compared to all treatments, except α-MEM+ and 1 ng/ml GDF-9. However, the concentration of 50 ng/ml GDF-9 showed higher cell proliferation and lower apoptosis than α-MEM+ and 1 ng/ml GDF-9 treatments. Culture of the ovarian tissue with LY294002 inhibited the activation of primordial follicles and reduced p-Akt immunostaining in both α-MEM+ and 50 ng/ml GDF-9 treatments. In addition, after culture with LY294002, the percentage of oocytes with nuclear p-FOXO3 was higher in 50 ng/ml GDF-9 than in the control medium (α-MEM+). In conclusion, after culture of ovine ovarian cortical slices, the addition of 50 ng/ml GDF-9 reduces follicular apoptosis and promotes granulosa cell proliferation likely through the involvement of phosphorylated Akt and FOXO3a.

Synthesis of controlled size starch nanoparticles (SNPs).

Starch nanoparticles (SNPs) are a promising choice for the strategic development of new renewable and biodegradable nanomaterials for novel biomedical and pharmaceutical applications when loaded with antibiotics or with anticancer agents as target drug delivery systems. The final properties of the SNPs are strongly influenced by the synthesis method and conditions being a controlled and monodispersed size crucial for these applications. The aim of this work was to synthesize controlled size SNPs through nanoprecipitation and microemulsion methods by modifying main operating parameters regarding the effect of amylose and amylopectin ratio in maize starches. SNPs were characterized by size and shape. SNPs from 59 to 118 nm were obtained by the nanoprecipitation method, registering the higer values when surfactant was added to the aqueous phase. Microemulsion method led to 35-147 nm sizes observing a higher particle formation capacity. The composition of the maize used influenced the final particle size and shape.

Epigallocatechin-3-gallate (EGCG) reduces apoptosis of preantral follicles through the phosphatidylinositol-3-kinase/protein kinase B (PI3K/AKT) signaling pathway after in vitro culture of sheep ovarian tissue.

The aims of this study were to analyze the effects of different concentrations of epigallocatechin-3-gallate (EGCG) on the primordial follicle survival and development after in vitro culture of ovarian tissue, and to verify the possible involvement of the phosphatidylinositol-3-kinase/protein kinase B (PI3K/AKT) pathway in the EGCG actions in the sheep ovary. Ovarian fragments were fixed for histological analysis (fresh control) or cultured in α-minimum essential medium alone (α-MEM+: control medium) or with different concentrations of EGCG (0.01; 0.1; 1; 10 or 100 µg/mL) for 7 days. Inhibition of PI3K activity was performed in fragments cultured with 1 µg/mL EGCG plus LY294002. Thereafter, immunohistochemistry was performed to evaluate the expression of cleaved caspase-3 and AKT phosphorylation (p-AKT). The results showed that 1 µg/mL EGCG maintained the follicular survival similar (P > 0.05) to that of the fresh control and higher (P < 0.05) than that of the α-MEM+ and other EGCG treatments. No difference (P > 0.05) in the follicular activation was observed. However, both follicle and oocyte diameters increased after in vitro culture with 1 µg/mL EGCG compared to other treatments (P < 0.05), except for 10 µg/mL EGCG (P > 0.05). After PI3K inhibition, there was an increase (P < 0.05) of the follicular apoptosis and a reduction of p-AKT immunolocalization. In conclusion, EGCG at 1 µg/mL reduces apoptosis of preantral follicles through the PI3K/AKT pathway after in vitro culture of sheep ovarian tissue.

MRI findings of the cervical spine in patients with mucopolysaccharidosis type VI: relationship with neurological physical examination.

AIM: To describe the magnetic resonance imaging (MRI) findings of the cervical spine of patients with mucopolysaccharidosis type VI (MPS VI) and correlate them with clinical manifestations. MATERIALS AND METHODS: This is a cross-sectional study involving 12 patients with MPS VI. A limited neurological examination was undertaken in each patient including Tinel's test, assessment of muscle tone, and the evaluation of deep tendon reflexes. Additionally, each patient underwent cervical spine MRI to evaluate platybasia, odontoid dysplasia, periodontoid soft-tissue thickening, spinal canal stenosis, myelopathy, basilar invagination, platyspondyly, and reduction of nasopharyngeal airway. RESULTS: Nine patients were male (75%). The average age was 12.5 (±3.5 years). Tinel's test was negative in all patients. No muscle tone abnormalities were observed. Approximately 48% of the tested reflexes were considered abnormal, 10 of which (8.3%) were pathological occurring in five different patients (41.6%). At MRI, all patients showed periodontoid soft-tissue thickening and cervical spinal stenosis; six showed spinal cord compression and two showed myelopathy. Odontoid hypoplasia and basilar invagination were observed in nine patients. All patients with cervical stenosis on MRI had abnormal reflexes; however, only two of the six patients with evidence of cord compression on MRI had abnormal reflexes on clinical examination. CONCLUSIONS: The present study of 12 patients with MPS VI demonstrated that a normal neurological examination cannot confidently exclude potential cord compression in patients with this condition. MRI may aid in the timely identification of cervical spine abnormalities, and potentially play a role in lessening morbidity and mortality in patients with MPS.

Angiotensin II receptor agonist antibodies are associated with microvascular damage in lupus nephritis.

Angiotensin II type 1 receptor agonist antibodies (AT1R-AAs) have been associated with hypertension, atherosclerosis and vascular inflammation in human diseases. The aim of the study was to evaluate the prevalence of AT1R-AAs in active lupus nephritis (LN) patients and their association with vascular damage. One hundred and seven active LN patients underwent a complete clinical examination, measurement of AT1R-AAs, ambulatory blood pressure monitoring, carotid intima-media thickness measurement and morphometric analysis of subintimal fibrosis and medial hyperplasia of the vessels in the kidney tissue. Plasma AT1R-AAs were positive in 58 (54.2%) patients. The Systemic Lupus Erythematosus Disease Activity Index 2000 (SLEDAI-2K) score, complement C3 and C4 levels and titers of anti-dsDNA antibodies were higher in the group with positive AT1R-AAs compared with those with negative AT1R-AAs. The AT1R-AA titers correlated with anti-dsDNA antibody titers and with complement C3 and C4 serum levels. In the kidney biopsy, the percentage of subintimal fibrosis and the area of medial hyperplasia were greater in the AT1R-AA-positive patients. No differences in arterial pressure, carotid intima-media thickness and response to therapy were detected. In conclusion, AT1R-AAs are prevalent in active LN patients and are associated with histologic features of microvascular damage.

Melatonin improves development, mitochondrial function and promotes the meiotic resumption of sheep oocytes from in vitro grown secondary follicles.

The aim of this study was to evaluate follicular survival and development of ovine isolated secondary follicles cultured in medium containing fixed or sequential concentrations of melatonin and further oocyte maturation. Isolated secondary follicles were cultured for 18 days in α-MEM+ alone (control) or with different concentrations of melatonin (100, 500 or 1000 pg/mL) or sequential concentrations of melatonin (Mel Seq: Day 6 = 100; Day 12 = 500; Day 18 = 1000 pg/mL). The percentages of morphologically normal follicles and antral cavity formation increased significantly in 1000 pg/mL melatonin compared to the other treatments. After 18 days, 1000 pg/mL melatonin (Mel 100) showed a greater (P < 0.05) follicular diameter than α-MEM+, 100 and 500 pg/mL melatonin. In addition, the concentration of 500 pg/mL melatonin showed a higher (P < 0.05) percentage of fully grown oocytes than α-MEM+, Mel 100 and Mel Seq treatments. After oocyte maturation, the levels of ROS were lower (P < 0.05) in 1000 pg/mL melatonin (Mel 1000) than in other treatments. Both Mel 1000 and Mel Seq treatments showed significantly higher levels of mitochondrial activity than other treatments. There were no significant differences between 500 and 1000 pg/mL melatonin regarding meiotic stages. In conclusion, the concentration of 1000 pg/mL melatonin maintains survival, promotes follicular development and increases the levels of active mitochondria after in vitro culture of sheep secondary follicles. Moreover, this concentration promotes the meiotic competence of oocytes and decreases the production of ROS during oocyte maturation.

Effects of melatonin on the in vitro growth of early antral follicles and maturation of ovine oocytes.

This study aimed to evaluate the effect of melatonin on the in vitro culture and maturation of isolated sheep early antral follicles. Isolated early antral follicles were cultured for 12 d in α-minimum essential medium (MEM+) alone (control) or α-MEM+ added with fixed different concentrations (100, 500, or 1,000 pg/mL) or a sequential concentration of melatonin (MelSeq; day 6 = 100; day 12 = 500 pg/mL). The percentage of morphologically normal follicles was higher (P < 0.05) in 500 pg/mL melatonin than the other treatments at 6 d. Mel 500 also showed a higher rate of fully grown oocytes (P < 0.05) than other treatments. After in vitro culture, reactive oxygen species (ROS) levels in oocytes were similar between Mel 500 and MelSeq, with both being lower (P < 0.05) than other treatments. Oocytes cultured in both Mel 500 and Mel 1000 showed glutathione peroxidase levels similar (P > 0.05) to the control group and higher (P < 0.05) than other treatments. Mitochondrial activity was similar (P > 0.05) among control, Mel 500, and Mel 1000 treatments. Mel 500 treatment presented a higher percentage of germinal vesicle breakdown oocytes than the control group and similar percentages to the other treatments. Follicles cultured in melatonin followed by oocyte maturation with the addition of 500 pg/mL melatonin in maturation medium showed increased (P < 0.05) levels of mitochondrial activity compared to α-MEM+ alone. In conclusion, the concentration of 500 pg/mL of melatonin promotes development and decreases ROS levels of ovine oocytes from in vitro grown early antral follicles. Moreover, melatonin increases mitochondrial activity and promotes the acquisition of meiotic competence of these oocytes.

Immunohistochemical localization of insulin-like growth factor-1 (IGF-1) in the sheep ovary and the synergistic effect of IGF-1 and FSH on follicular development in vitro and LH receptor immunostaining.

This study analyzed IGF-1 protein immunostaining in sheep ovaries, the effect of IGF-1 alone or associated with FSH on the culture of secondary follicles, and the immunostaining of LHR protein in antral follicles before and after culture. Ovaries were collected for IGF-1 protein analysis. In experiment 1, secondary follicles were cultured in α-MEM+ (control) or α-MEM+ supplemented with IGF-1 (10, 50 or 100 ng/mL). In experiment 2, follicles were cultured in the same media of experiment 1 plus 750 ng/mL FSH. Moreover, LHR immunostaining was analyzed in fresh antral follicles and after culture in 50 ng/mL IGF-1 + FSH. The IGF-1 protein was immunolocalized in oocytes from all stages of follicle development and in the granulosa cells from secondary and antral follicles. IGF-1 did not influence (P > 0.05) follicular viability and growth (experiment 1). However, in experiment 2, 50 ng/mL IGF-1 + FSH stimulated oocyte growth (P < 0.05) and LHR immunostaining in antral follicles. Control medium, 10 or 50 ng/mL IGF-1 + FSH showed similar levels of reactive oxygen species, glutathione and active mitochondria (P > 0.05). In conclusion, the IGF-1 protein is present in all ovarian follicle stages in sheep. Moreover, the association between 50 ng/mL IGF-1 and FSH has a synergistic effect in vitro, increasing the percentage of fully grown oocytes and the intensity of immunostaining of LHR protein in oocytes and granulosa cells of cultured antral follicles.