Oncofertility case report: live birth 10 years after oocyte in vitro maturation and zygote cryopreservation.

PURPOSE: This study aims to report a case of urgent fertility preservation in an oncological patient with collection of immature oocytes in the absence of ovarian stimulation that, through in vitro maturation (IVM), followed by ICSI and cryopreservation of zygotes resulted, 10 years later, in the live birth of a healthy baby. METHODS: In September 2008, our clinic performed IVM in a 32-year-old woman diagnosed with a ductal invasive carcinoma with positive estradiol receptors, negative progesterone receptors and positive human epidermal growth factor receptor 2. The retrieval of immature oocytes was performed in the absence of ovarian stimulation after a simple mastectomy and prior to any chemotherapy treatment. The compact cumulus-oocyte complexes (COCs) collected were placed in Lag medium for 2 h, followed by incubation in IVM medium, supplemented with heat inactivated patient serum, recombinant FSH, and recombinant LH. After 30 h in culture, cumulus cells were removed, the metaphase II oocytes were microinjected, and the zygotes obtained were cryopreserved. In 2017, the zygotes were thawed and cultured until day 3. One embryo was transferred and the other cryopreserved. RESULTS: Four compact COCs were collected and subjected to IVM. Two oocytes reached metaphase II and were microinjected. Two zygotes were obtained and were cryopreserved at the two pronuclear stage. Approximately 9 years later, the two zygotes were thawed and cultured until day 3. An embryo with 10 cells was transferred and implanted, resulting in the birth of a healthy baby. CONCLUSIONS: In cases where urgency to start adjuvant therapy requires immediate oocyte collection, IVM may be the only option to obtain fully competent mature oocytes allowing for effective preservation of the reproductive potential.

Improving crunchiness and crispness of fried squid rings through innovative tempura coatings: addition of alcohol and CO2 incubation.

This study aimed to determine the effect of different uncommon tempura formulations (incubated with CO2 and with added ethanol) on physical, chemical and sensory characteristics of fried coated squids, immediately after frying and also after 48 h of refrigeration storage and subsequent oven reheating. Ethanol addition led to lower levels of moisture and higher of fat in the fried coating, regardless the use of CO2. There were no difference in instrumental colour parameters among all four battered samples. Ethanol added tempuras showed higher crispness and lower maximum force than their water counterparts in an instrumental texture evaluation. Fried squids coated with ethanol added tempuras were rated as crispier and crunchier, even after 2 days of storage and further reheating. Overall, partial substitution of water by ethanol appears as an interesting strategy to increase crispness in tempura coated fried products, especially when the products are stored and reheated before consumption.

Cleaning cassava genotypes infected with cassava frogskin disease via in vitro shoot tip culture.

This study aimed to develop a methodology for eliminating cassava frogskin disease (CFSD) from in vitro shoot tip culture by associating thermotherapy and tetracycline. Cuttings from different accessions (BGM0232, BGM0315, BGM0464, BGM584, BGM0841, and BGM1342), infected with CFSD according to visual inspection of the disease symptoms, were used for cleaning. To verify the absence of other diseases, the plants were indexed for Cassava common mosaic virus - CsCMV (by ELISA) and Cassava vein mosaic virus - CsVMV (by polymerase chain reaction, PCR), proving that the accessions were free of these viruses, except for BGM0315 and BGM0464, which were infected with CsVMV. Subsequently, the cuttings were submitted to different tetracycline concentrations for 3 min, and then subjected to thermotherapy under different temperatures (35°, 38°, 40°, 45°, and 55°C). Shoots of 2 cm were harvested, and their surfaces were sterilized in a laminar flow chamber. Subsequently, the shoot tips of different sizes were removed (0.2, 0.4, 0.5, and 1.0 mm) for inoculation in a culture medium with tetracycline at the same concentrations in which the cuttings were dipped. After 60 days of cultivation, the explants were transferred to a multiplication medium without antibiotics. Thirty days after the transfer, the viability of the regenerated plants was evaluated, which were then acclimatized for 70 days in a greenhouse and transferred to the field. After 7 months, a visual analysis of the symptomatic roots and a PCR analysis were held to prove the elimination of CFSD and CsVMV from the accessions infected with these viruses (BGM0315 and BGM0464), respectively. Most of the treatments resulted in 100% cleaning of CFSD-infected plants. From accessions that were also infected with CsVMV, only 2% of the plants remained infected, also demonstrating the cleaning efficiency of this protocol for this disease.

Effect of amniotic membrane proteins in human cancer cell lines: an exploratory study.

Human amniotic membrane (hAM) has recently drawn attention as an upcoming anti-cancer therapy. Regarding the strategies which have already investigated, little is known about hAM protein extracts (hAMPE) effect on cancer. So, this work aims to study the effect of hAMPE in metabolic activity of several human cancer cell lines. hAMPE were mechanically obtained, thus avoiding the effect of detergents and other reagents commonly used in protein extraction under the cell lines studied. After quantification of proteins in hAMPE, their effect on the metabolic activity of 21 human cancer cell lines was assessed by 3-(4,5-dimethylthia-zolyl-2)2,5-diphenyltetrazolium bromide (MTT) assay. Our results indicate that there is an inhibition of metabolic activity until 25 and 50% in two and seven cell lines, respectively. Five cell lines proved to be very sensitive to hAMPE, being its metabolic activity more than 50% inhibited. Our results show that hAMPE can inhibit the metabolic activity of some human cancer cell lines. However, research about this cell line-dependent response to hAMPE becomes indispensable.

Characterisation of colistin resistance in Gram-negative microbiota of pregnant women and neonates in Nigeria.

A mobile colistin resistance gene mcr was first reported in 2016 in China and has since been found with increasing prevalence across South-East Asia. Here we survey the presence of mcr genes in 4907 rectal swabs from mothers and neonates from three hospital sites across Nigeria; a country with limited availability or history of colistin use clinically. Forty mother and seven neonatal swabs carried mcr genes in a range of bacterial species: 46 Enterobacter spp. and single isolates of; Shigella, E. coli and Klebsiella quasipneumoniae. Ninety percent of the genes were mcr-10 (n = 45) we also found mcr-1 (n = 3) and mcr-9 (n = 1). While the prevalence during this collection (2015-2016) was low, the widespread diversity of mcr-gene type and range of bacterial species in this sentinel population sampling is concerning. It suggests that agricultural colistin use was likely encouraging sustainment of mcr-positive isolates in the community and implementation of medical colistin use will rapidly select and expand resistant isolates.

Amniotic membrane: from structure and functions to clinical applications.

Amniotic membrane (AM) or amnion is a thin membrane on the inner side of the fetal placenta; it completely surrounds the embryo and delimits the amniotic cavity, which is filled by amniotic liquid. In recent years, the structure and function of the amnion have been investigated, particularly the pluripotent properties of AM cells, which are an attractive source for tissue transplantation. AM has anti-inflammatory, anti-bacterial, anti-viral and immunological characteristics, as well as anti-angiogenic and pro-apoptotic features. AM is a promoter of epithelialization and is a non-tumorigenic tissue and its use has no ethical problems. Because of its attractive properties, AM has been applied in several surgical procedures related to ocular surface reconstruction and the genito-urinary tract, skin, head and neck, among others. So far, the best known and most auspicious applications of AM are ocular surface reconstruction, skin applications and tissue engineering. However, AM can also be applied in oncology. In this area, AM can prevent the delivery of nutrients and oxygen to cancer cells and consequently interfere with tumour angiogenesis, growth and metastasis.

Antibiotic resistance genes in the gut microbiota of mothers and linked neonates with or without sepsis from low- and middle-income countries.

Early development of the microbiome has been shown to affect general health and physical development of the infant and, although some studies have been undertaken in high-income countries, there are few studies from low- and middle-income countries. As part of the BARNARDS study, we examined the rectal microbiota of 2,931 neonates (term used up to 60 d) with clinical signs of sepsis and of 15,217 mothers screening for blaCTX-M-15, blaNDM, blaKPC and blaOXA-48-like genes, which were detected in 56.1%, 18.5%, 0% and 4.1% of neonates' rectal swabs and 47.1%, 4.6%, 0% and 1.6% of mothers' rectal swabs, respectively. Carbapenemase-positive bacteria were identified by MALDI-TOF MS and showed a high diversity of bacterial species (57 distinct species/genera) which exhibited resistance to most of the antibiotics tested. Escherichia coli, Klebsiella pneumoniae and Enterobacter cloacae/E. cloacae complex, the most commonly found isolates, were subjected to whole-genome sequencing analysis and revealed close relationships between isolates from different samples, suggesting transmission of bacteria between neonates, and between neonates and mothers. Associations between the carriage of antimicrobial resistance genes (ARGs) and healthcare/environmental factors were identified, and the presence of ARGs was a predictor of neonatal sepsis and adverse birth outcomes.

Training and detraining effects on functional fitness after a multicomponent training in older women.

BACKGROUND: Several studies have been carried out in order to evaluate the potential influence of increased physical activity on the health, biological ageing and functional ability of the elderly. However, only limited information is available on the effects of multicomponent training and detraining on functional performance. OBJECTIVE: The purpose of the present study was to investigate the effect of 8-month multicomponent training and 3-month detraining on the functional fitness of older women. METHODS: Fifty-seven women were randomly assigned to an exercise (n = 32; 68.4 +/- 2.93 years) or a control group (n = 25; 69.6 +/- 4.20 years). The training program consisted of 2 sessions per week of aerobic, strength, balance and flexibility exercises. The functional fitness test battery was performed to assess the physical parameters associated with independent functioning in older adults. RESULTS: No significant changes were observed in body mass index and cardiovascular endurance as a result of the exercise training. Training induced significant (p < 0.05) improvements in chair stand (27.3%), arm curl (17.4%), chair sit-and-reach (17.4%), up-and-go (11%) and back scratch (14.5%) tests. However, both upper and lower body strength and upper and lower flexibility declined significantly after detraining in the exercise group. CONCLUSION: The results of this study highlight the negative effects of interrupting exercise on several physical parameters of functional fitness.

Novel nanocomposite coatings with dispersed organic nanoparticles for solar absorbers.

A natural pigment (Anthocyanin extracted from Sambucus nigra L. fruit) was incorporated by dipping sputtered titanium oxide films into a pigment solution. The pigment solution has red colour and an absorption peak located in the wavelength range of 520 to 540 nm. Titanium oxide thin films were prepared by reactive DC magnetron sputtering and reactive pulsing DC magnetron sputtering from a Titanium target in O2 + Ar atmosphere onto glass slides. Pigment incorporation in thin films was found dependent of both morphology and composition, which depends on deposition parameters, like pulsing frequency and oxygen flow rate. The incorporation of this pigment in TiOx thin films increases the final solar absorption and decreases the final average roughness of these films. The effect in solar absorption is higher in films prepared by DC and pulsed DC at pulsing frequency of 200 kHz.

Breast cancer patients with micrometastases in sentinel lymph nodes: differences considering additional metastatic lymph nodes.

AIMS: Characterization of breast cancer patients with micrometastases in sentinel lymph node (SLN) and establish differences between micrometastatic breast cancers with additional metastatic lymph nodes (LNS) versus no other lymph node invasion. METHODS: Analysis of 30 breast cancers, N1mi or pN0(i+), diagnosed and treated in our department from July 2000 to July 2008. RESULTS: Micrometastases in SLNs were found in 30 patients. Complete axillary dissection revealed other metastatic LNs in 24%. Concerning breast cancers with additional LN invasion versus no other LN invasion, tumors located in the superior-external quadrant were more frequent in the former group. Other characteristics as clinical presentation, histological subtype, focality, cytonuclear grade, hormone receptors and Her2 expression were not significantly different in either group. Regarding SLN invasion, the presence of at least two micrometastatic foci were significantly more relevant in patients with other metastatic LN invasion (p < 0.01). Micrometastases diagnosed only after immunohistochemistry (IHC) were exclusively found in patients without other LN invasion, reaching statistical significance (p < 0.05). CONCLUSIONS: Complete axillary dissection revealed additional LN invasion in 24% of patients with micrometastases in the SLN. Tumors with additional LN invasion were more frequently found in the superior external quadrant and SLNs harbored at least two micrometastatic foci. Micrometastases diagnosed exclusively by IHC techniques were more relevant in cases without additional lymph node invasion.

What is the diagnostic value of nipple discharge cytology and galactography in detecting duct pathology?

The present study aimed to evaluate the diagnostic value of nipple discharge (ND) cytology and galactography. Ninety-four patients submitted to duct excision, representing a total of 98 duct excisions, were retrospectively analyzed from January 1997 to May 2007. Histology of ducts excised revealed 35% duct ectasia (DE), 31% duct papilloma (DP), 20% potential malignant transforming lesions (PMTL), 6% breast cancer (BC), 1% adenoma and 6% normal breast tissue. Cytology had a sensibility and specificity in detecting duct pathology of, respectively, 40% and 61.3%, a positive predictive value (PPV) of 53.8% and a negative predictive value (NPV) of 47.5%. Concerning malignant and PMTL, cytology had a sensibility and specificity of, respectively, 46.2% and 62.3%, a PPV of 25% and a NPV of 82.5%. Breast cancer was never suggested by positive cytology. Galactography had a sensibility and specificity in detecting duct pathology of, respectively, 77.4% and 29.2%, a PPV of 58.5% and a NPV of 50%. For malignant and PMTL, galactography had a sensibility and specificity of, respectively, 80% and 26.7%, a PPV of 19.5% and a NPV of 85.7%. However, galactography never suggested a diagnosis of cancer. Cytology and galactography performed together in 49% showed a low Kappa Index (KI < 1), allowing us to conclude that cytology and galactography detect different pathologies. Cytology showed a poor performance in predicting histological diagnosis in face of ND. Galactography had a good sensibility in excluding malignant lesions and PMTL. Galactography was significantly more sensitive for duct pathology but cytology was more specific for malignant lesions and PMTL.

Oxidative Stress, DNA, Cell Cycle/Cell Cycle Associated Proteins and Multidrug Resistance Proteins: Targets of Human Amniotic Membrane in Hepatocellular Carcinoma.

The anticancer effects of human amniotic membrane (hAM) have been studied over the last decade. However, the action mechanisms responsible for these effects are not fully understood until now. Previously results reported by our team proved that hAM is able to induce cytotoxicity and cell death in hepatocellular carcinoma (HCC), a worldwide high incident and mortal cancer. Therefore, this experimental study aimed to investigate the cellular targets of hAM protein extracts (hAMPE) in HCC through in vitro studies. Our results showed that hAMPE is able to modify oxidative stress environment in all HCC cell lines, as well as its cell cycle. hAMPE differently targets deoxyribonucleic acid (DNA), P21, P53, ß-catenin and multidrug resistance (MDR) proteins in HCC cell lines. In conclusion, hAMPE has several targets in HCC, being clear that the success of this treatment depends of a personalized therapy based on the biological and genetic characteristics of the tumor.

Selective cytotoxicity and cell death induced by human amniotic membrane in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) has a worldwide high incidence and mortality. For this reason, it is essential to invest in new therapies for this type of cancer. Our team already proved that human amniotic membrane (hAM) is able to inhibit the metabolic activity of several human cancer cell lines, including HCC cell lines. Taking into account the previously performed work, this experimental study aimed to investigate the pathways by which hAM protein extracts (hAMPEs) act on HCC. Our results showed that hAMPE reduce the metabolic activity, protein content and DNA content in a dose- and time-dependent manner in all HCC cell lines. This therapy presents selective cytotoxicity, since it was not able to inhibit a non-tumorigenic human cell line. In addition, hAMPE induced cell morphology alterations in all HCC cell lines, but death type is cell line dependent, as proved by in vitro and in vivo studies. In conclusion, hAMPE have a promising role in HCC therapy, since it is capable of inducing HCC cytotoxicity and cell death.

Diurnal blood pressure variation in progressive autonomic failure.

To investigate the role of the autonomic nervous system (ANS) in the generation of the circadian blood pressure (BP) variation, the degree of impairment of the ANS was related to the results of ambulatory BP recordings in 212 patients with progressive autonomic failure due to familial amyloid polyneuropathy. On the basis of BP and/or heart rate (HR) responses to the Valsalva maneuver, 60 degrees head-up tilting, deep-breathing tests, and plasma norepinephrine levels, 4 groups of patients were distinguished. In all patients and in 38 age-matched control subjects, ambulatory BP was monitored. Patients of group I (n=40, aged 32+/-3 y), with no evidence yet of impairment of their ANS, had circadian BP and HR variations indistinguishable from controls. Patients of group II (n=41, aged 34+/-5 y) had a variable degree of impairment of their parasympathetic ANS, but their sympathetic ANS was still intact. Twenty-four-hour HR was higher in these patients than in controls (88+/-11 versus 78+/-7 bpm, P<0.01). Their circadian HR variation was maintained, but their circadian BP variation was diminished (10+/-6/11+/-4 versus 17+/-6/16+/-4 mm Hg in controls, P<0.01) because of an attenuation of the nocturnal BP decline. Patients of group III (n=69, aged 36+/-6 y), with parasympathetic failure and intermediate sympathetic dysfunction, had a blunted diurnal BP variation, whereas patients of group IV (n=62, aged 38+/-6 y), with parasympathetic failure and severe sympathetic dysfunction, had an absent diurnal BP variation. In patients of groups III and IV, a decrease in daytime BP accounted for the blunted circadian BP variation. This extensive study in progressive autonomic failure confirms the important role of the ANS in the generation of circadian BP variation. For the maintenance of a normal circadian BP pattern, not only an intact sympathetic but also an intact afferent parasympathetic ANS is a prerequisite.

Role of sympathetic nervous system in cyclosporine-induced rise in blood pressure.

To clarify the role of the sympathetic nervous system in the development of cyclosporine A (CsA)-induced rise in blood pressure (BP), the effects of CsA on 24-hour ambulatory BP (ABP) were studied in patients with familial amyloid polyneuropathy (FAP) who underwent a liver transplantation. On the basis of autonomic function tests, patients with absent or mild-to-moderate sympathetic damage (Group A, n=11, age 29 to 43 years, disease duration 2 to 6 years) and patients with severe sympathetic damage (Group B, n=9, age 27 to 38 years, disease duration 3 to 9 years) were identified. Both groups were followed for 1 year. The daily doses of CsA and the CsA whole blood trough levels between the groups did not differ. Pretransplantation values of daytime and nighttime ABP were, respectively, 117+/-8/76+/-7 mm Hg and 108+/-12/68+/-9 mm Hg in group A and 107+/-6/66+/-4 mm Hg (P<0.05 group A versus group B) and 102+/-6/62+/-4 mm Hg in group B. In response to CsA, BP increased in all patients, but more so in patients of group B than in patients of group A. One year after transplantation, daytime and nighttime ABP had increased by 6+/-9/3+/-11% and 12+/-10/14+/-14% in group A and by 12+/-6/13+/-10% (P<0.05) and 21+/-11/27+/-21% (P<0.01) in group B. In both groups, the increase in nighttime ABP was greater than the increase in daytime ABP, which resulted in an attenuation or, even, a reversal of the diurnal BP rhythm. Because the rise in BP was greater in patients with more advanced sympathetic dysfunction, the sympathetic nervous system appears to counteract the CsA-induced rise in BP rather than causing it. This implies involvement of factors other than sympathetic activation in the pathogenesis of CsA-induced rise in BP in patients with familial amyloid polyneuropathy.

High altitude-related neurocardiogenic syncope.

This report presents a well-documented link between vasodepressor syncope, a marked increase of LF/HF ratio response to orthostasis (sympathetic dominance), and the efficacy of beta blockers in preventing orthostatic symptoms and absence of tilt-induced syncope, with normalization of the LF/HF ratio response.

Nocardial keratitis following myopic keratomileusis.

PURPOSE/METHODS: Interface opacities were detected in a patient who underwent uncomplicated myopic keratomileusis. RESULTS AND DISCUSSION: Nocardia asteroides keratitis was confirmed by microbiologic work-up and guided the correct treatment. The eye recovered 20/60 spectacle-corrected visual acuity and had a residual stromal scar.

Cyclosporine versus tacrolimus in kidney transplantation: are there differences in nephrotoxicity?

Cyclosporine and tacrolimus, two calcineurin inhibitors, show different side effects and toxicities. The data concerning their nephrotoxicity are few and conflicting. A retrospective study was performed in 2 groups of renal transplant recipients treated with cyclosporine or tacrolimus to evaluate graft function and side effects. All patients had completed at least 6 months of follow-up before inclusion in the study. Group I included 10 patients who were converted from cyclosporine to tacrolimus, due to cosmetic problems or due to chronic graft dysfunction with creatinine values <3 mg/dL. After conversion, there was a significant reduction in creatinine values (from 2.43 +/- 1.21 to 1.86 +/- 0.72 mg/dL; P =.023) and an improvement in creatinine clearance (from 47.5 +/- 19.2 to 56.1 +/- 18.9 mL/min; P =.047). The lipid profile did not change, but there was a trend to better blood pressure control with less antihypertensive drugs. Group II compared 2 subgroups of patients receiving kidneys from the same donor, one treated with cyclosporine and the other with tacrolimus. Tacrolimus patients showed better renal function; namely, creatinine was 1.15 +/- 0.27 versus 1.44 +/- 0.33 mg/dL (P =.029) and creatinine clearance was 87.7 +/- 27.1 versus 60.3 +/- 25.9 mL/min (P =.043). Lipid and blood pressure values were not different between the 2 subgroups, but tacrolimus patients tended to need a lower number of antihypertensive medications. The incidence of de novo diabetes mellitus was approximately 20% among patients using tacrolimus. We concluded that tacrolimus may be less nephrotoxic than cyclosporine. Tacrolimus patients showed better graft function and easier blood pressure control, but a high incidence of posttransplantation diabetes mellitus.

Spectral analysis of heart rate variability in athletes.

The objectives of the study were to characterize power spectrum pattern of the heart rate variability and assessment of the relative contributions of sympathetic and parasympathetic cardiac nervous system control in athletes. Thirty-three male athletes, swimming (1), canoeing (10), cycling (6), athletics (4), football (3), roller-skating (2) and volleyball (7) aged 23.4 +/- 5.5 years, with a mean athletic level of 18 hours/week (8-45) and 33 sedentary healthy control subjects were included. Ecg signals were recorded after a period of 15 minutes in supine rest with controlled breathing at 15 cycles/min. Signal acquisition was done at 300 samples/sec. From 512 consecutive heart beats, we calculated mean average, standard deviation, maximum and minimum R-R intervals and, after computing the fast Fourier transform, total spectrum power, low frequency (LF), high frequency (HF) components and its ratio (LF/HF). The average R-R interval was 987.7 +/- 168.8 ms and 762.7 +/- 125.3 ms, the variance was 5.44 and 2.51 ms2 and ratio of R-R interval maximum/minimum (E/I ratio) 1.53 +/- 0.16 and 1.41 +/- 0.16, respectively for athletes and control group. Differences between groups were significant (p < 0.01) for all parameters, with higher variability in the athletes. Both spectral bands (LF and HF) had higher power in athletes (LF = 925 +/- 920 and HF = 2258 +/- 2349 ms2) than in the control group (LF = 442 +/- 446 and HF = 1179 +/- 1542 ms2) (p < 0.01). There were no significant differences for LF/HF ratio, or normalized LF (LF%) or normalized HF (HF%) between groups.(ABSTRACT TRUNCATED AT 250 WORDS)