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.

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.

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.

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.

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.

Better outcomes of peritoneal dialysis in diabetic patients in spite of risk of loss of autonomy for home dialysis.

INTRODUCTION: Diabetes mellitus is a leading cause of chronic renal failure, challenging therapy strategies. Patients with diabetes may benefit from peritoneal dialysis (PD) but higher technique failure is feared. Our purpose was to critically evaluate clinical outcomes of this modality in diabetics, in order to find quality improvement strategies in these risk patients. METHODS: A registry-based study of 432 incident patients, 23% with diabetes, from a university hospital PD program was performed. Traditional methods (Kaplan-Meier, Cox models) and innovative survival analysis, taking competing risks into account, were performed, as well as exploring the trends in cohorts according to the decade of PD start. RESULTS: In spite of the detrimental effect of diabetes in patient survival compared to non-diabetics (77%, 52% vs 86%, 71%, at 2 and 4 years, respectively; p < 0.0001) the hazard ratio (HR) for death decreased in the more contemporary cohort (0.303, 95% confidence interval (CI) 0.150 - 0.614, p < 0.001). It is noteworthy that diabetes was not associated with lower technique survival: 74%, 51% vs 79%, 57%, at 2 and 4 years, respectively (p = not significant (NS)). On multivariate analysis, diabetes was an independent predictor for mortality, but not for technique failure. The hazard ratio (HR) for technique failure also decreased in the more recent cohort (0.566, 95% CI 0.348 - 0.919, p = 0.021). Among reasons for transfer to hemodialysis, proportion of ultrafiltration failure was similar between groups (26% vs 22%, p = NS), but drop-out due to loss of autonomy occurred more in the group with diabetes (23% vs 5%, p = 0.004), mainly due to ischemic stroke. The hospitalization rate was also higher in diabetic patients (1.39 vs 0.84 episodes per patient-year (E/PY), p = 0.004) but the peritonitis rate was not increased (0.53 vs 0.61 E/PY, p = NS). CONCLUSION: PD was an effective long-term renal replacement therapy in diabetics, without higher rates of technique failure, ultrafiltration failure or peritonitis. Better outcomes were achieved in the contemporary cohort. The menace of autonomy loss due to stroke and higher hospitalization rates enhance the need for assisted PD strategies and better control of comorbidities.

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.

Electre Tri-C, a multiple criteria decision aiding sorting model applied to assisted reproduction.

OBJECTIVE: The aim of this paper is to apply an informatics tool for dealing with a medical decision aiding problem to help infertile couples to become parents, when using assisted reproduction. METHODS: A multiple criteria decision aiding method for sorting or ordinal classification problems, called Electre Tri-C, was chosen in order to assign each couple to an embryo-transfer category. The set of categories puts in evidence a way for increasing the single pregnancy rate, while minimizing multiple pregnancies. The decision aiding sorting model was co-constructed through an interaction process between the decision aiding analysts and the medical experts. RESULTS: According to the sample used in this study, the Electre Tri-C method provides a unique category in 86% of the cases and it achieves a sorting accuracy of 61%. Retrospectively, the medical experts do agree that some of their judgments concerning the number of embryos to transfer back to the uterus of the woman could be different according to these results. The current ART methodology achieves a single pregnancy rate of 47% and a twin pregnancy rate of 14%. Thus, this informatics tools may play an important role for supporting ART medical decisions, aiming to increase the single pregnancy rate, while minimizing multiple pregnancies. LIMITATIONS: Building the set of criteria comprises a part of arbitrariness and imperfect knowledge, which require time and expertise to be refined. Among them, three criteria are modeled by means of a holistic classification procedure by the medical experts.

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.

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.

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.