Biologically synthesized silver nanoparticles as potent antibacterial effective against multidrug-resistant Pseudomonas aeruginosa.

Pseudomonas aeruginosa is one of the most worrisome infectious bacteria due to its intrinsic and acquired resistance against several antibiotics and the recalcitrance of its infections; hence, the development of novel antimicrobials effective against multidrug-resistant P. aeruginosa is mandatory. In this work, silver nanoparticles obtained by green synthesis using a leaf extract and fungi were tested against a battery of clinical strains from cystic fibrosis, pneumonia and burnt patients, some of them with multidrug resistance. Both nanoparticles showed a potent antibacterial effect, causing severe damage to the cell wall, membrane and DNA, and inducing the production of reactive oxygen species. Moreover, the nanoparticles derived from fungi showed synergistic antibacterial effects with the antibiotics meropenem and levofloxacin for some clinical strains and both kinds of nanoparticles were nontoxic for larvae of the moth Galleria mellonella, encouraging further research for their implementation in the treatment of P. aeruginosa infections.

Influence of minimally invasive endodontic access cavities on root canal shaping and filling ability, pulp chamber cleaning and fracture resistance of extracted human mandibular incisors.

AIM: To evaluate the influence of the location and design of endodontic access cavities on root canal shaping and filling ability, pulp chamber cleaning and fracture resistance of extracted human mandibular incisors. METHODOLOGY: After pre-selection using periapical radiographs, forty extracted intact human mandibular incisors were scanned in a micro-computed tomographic device. The teeth were matched based on similar anatomical features of the canals and assigned to four experimental groups (n = 10) according to the endodontic access cavity and root canal preparation protocol: traditional/TRUShape (T/TRU); traditional/MTwo (T/MT); minimally invasive/TRUShape (MI/TRU); and minimally invasive/MTwo (MI/MT). The samples were scanned after root canal instrumentation and filling procedures. The parameters evaluated were as follows: volume and area of the root canal, noninstrumented canal areas, canal transportation and centring ratio, accumulated hard tissue debris, voids in root canal fillings and remnants of root canal filling materials in the pulp chamber. After root canal filling and cavity restoration procedures, the samples were submitted to a fracture resistance test. Data were statistically analysed using Shapiro-Wilk, one-way anova and Bonferroni tests with a significance level of 5% (α = 0.05). RESULTS: There was no difference regarding all parameters evaluated before and after root canal preparation (volume and area of the root canal, noninstrumented canal areas, canal transportation and centring ratio, and accumulated hard tissue debris) amongst the groups (P > 0.05). MI/TRU and MI/MT groups were associated with significantly more voids in root canal fillings when compared to the T/TRU and T/MT groups (P < 0.05). Percentage of root canal filling material remnants in the pulp chamber after cleaning procedures and mean fracture resistance values were not significantly different amongst the four experimental groups (P > 0.05). CONCLUSIONS: The location and design of the endodontic access cavity did not impact on root canal preparation nor resistance to fracture of extracted mandibular incisors, regardless of the instrument used. Minimally invasive access cavities were associated with significantly more voids in root canal fillings.

Room-temperature, atmospheric-pressure microsputtering of dense, electrically conductive, sub-100 nm gold films.

In this study, we investigate the effects of deposition parameters (i.e. gas flow rates, bias voltages, and nozzle-to-substrate separation) on the microstructure of <100 nm thick gold deposits made with a room-temperature, atmospheric-pressure, ion-drag microsputterer. Without resorting to the use of vacuum or substrate heating, optimization of the printing process yields dense, continuous deposits (96.5% coverage) with low electrical resistivity (45 µΩ cm). Using statistical analysis, we developed a simple model that provides insight into the dynamics of such a printing method; based on this model, we identify electrostatic effects as the most important factor that influences the deposition process.

Cementum protein 1 transfection does not lead to ultrastructural changes in nucleolar organization of human gingival fibroblasts.

BACKGROUND AND OBJECTIVE: Transfection of cementum protein 1 (CEMP1) into human gingival fibroblasts (HGFs) notably increases cell metabolism and results in overexpression of molecules related to biomineralization at transcriptional and protein levels. Therefore, HGF-CEMP1 cells are considered as putative cementoblasts. This represents a significant advance in periodontal research because cementum neoformation is a key event in periodontal regeneration. In addition, it is well known that important changes in cell metabolism and protein expression are related to nucleolar structure and the function of this organelle, which is implicated in ribosome biogenesis. The aim of this study was to determine the effect of transfecting CEMP1 gene in human HGF on the ultrastructure of the nucleolus. MATERIAL AND METHODS: Cells were processed using the conventional technique for transmission electron microscopy, fixed with glutaraldehyde, postfixed with osmium tetraoxide, and embedded in epoxy resin. Semi-thin sections were stained with Toluidine blue and observed by light microscopy. Thin sections were stained with uranyl acetate and lead citrate. For ribonucleoprotein detection, the staining method based on the regressive effect of EDTA was used. In addition, the osmium ammine technique was used for specific staining of DNA. RESULTS: The results obtained in this study suggest that transfection of CEMP1 into HGFs does not produce changes in the general nucleolar ultrastructure because the different components of the organelle are present as fibrillary centers, and dense fibrillar and granular components compared with the control. CONCLUSION: The transfection of CEMP1 into HGFs allows these cells to perform cementoblast-like functions without alteration of the ultrastructure of the nucleolus, evaluated by the presence of the different compartments of this organelle involved in ribosomal biogenesis.

Retracted: Micro-computed tomography evaluation of apical transportation and centring ability of Reciproc and WaveOne systems in severely curved root canals.

The following article from International Endodontic Journal, 'Micro-computed tomography evaluation of apical transportation and centring ability of Reciproc and WaveOne systems in severely curved root canals' by D. A. de Meireles, T. C. C. A. de Brito, A. A. F. Marques, A. D. B. Garrido, L. F. R. Garcia & E. C. Sponchiado Jr, published online on 5 February 2015 in Wiley Online Library (wileyonlinelibrary.com), has been retracted by agreement between the authors, the journal Editor in Chief, Prof. Paul Dummer, and John Wiley & Sons Ltd. The retraction has been agreed due to the use of techniques for crucial measurements in canal shaping and a lack of clarity regarding the measuring methodology. The use of inadequate measuring methodologies makes the findings of the paper invalid.

Evaluation of Toll-like receptor and adaptor molecule polymorphisms for susceptibility to tuberculosis in a Colombian population.

Immunological studies have supported the idea that innate immunity is critical for the control of Mycobacterium tuberculosis (Mtb) infection in humans. Despite the overwhelming evidence showing the critical role of Toll-like receptors (TLRs) in the in vitro recognition of Mtb, the in vivo significance of individual TLRs has been more difficult to demonstrate consistently. We were interested in examining the role of genes of TLRs and molecules involved in their signalling cascades, and a case-control study was designed to test the association of polymorphisms of these innate immune genes with pulmonary tuberculosis (TB) in a Colombian population. In this study, we did not find an association with TLR2, TLR4, TLR9, MyD88 or MAL/TIRAP polymorphic variants. These findings suggest that those genes are not involved as risk factors for pulmonary TB in our population.

Association between serum calcium levels and the presentation of postpartum endometritis in housed dairy cows.

The present work aims to identify the minimum threshold of serum calcium (SC) values in cows during the first week postpartum and evaluate their relationship with the presentation of endometritis in housed dairy cows. In this prospective longitudinal observational study, 467 cows from 3 farms in Lima-Peru were studied. Blood samples were collected from cows during the first week postpartum. Endometritis was diagnosed on day 35 ± 3 days postpartum by evaluation of vaginal discharge samples. The samples were obtained using the MetricheckTM device (Simcro, New Zealand). Two cut-off points were used to diagnose endometritis: a) endometritis metricheck score ≥ 3, and b) endometritis metricheck score ≥ 2. In the univariate model that considers a SC threshold of 5.25 to 8.75 mg/dL (1.31 to 2.18 mmol/L), a significant relationship (p < 0.05) was found for endometritis metricheck score ≥ 3, while no significant relationship was found (p = 0.12) with endometritis metricheck score ≥ 2. In both cases, the optimal SC threshold for the appearance of endometritis was determined to be a value ≥7.0 mg/dL (≥1.75 mmol/L). In the logistic regression models (parity, calving season, calcium level, and their interaction), only two variables were retained, parity and calcium level (p < 0.10). It was found that the probability of having endometritis metricheck score ≥ 3 was 1.9 (95% CI: 1.1 to 3.1), whereas the probability of having endometritis metricheck score ≥ 2 was 1.5 (95% CI: 1.0 to 2.5) in cows with calcium levels <7.0 mg/dL (<1.75 mmol/L). In conclusion, cows with calcium levels <7.0 mg/dL (<1.75 mmol/L) have a 1.9-fold greater risk of developing endometritis metricheck score ≥ 3 in the first week postpartum.

Cross-Cultural Measurement Invariance in the Personality Inventory for DSM-5✰.

The validity of cross-cultural comparisons of test scores requires that scores have the same meaning across cultures, which is usually tested by checking the invariance of the measurement model across groups. In the last decade, a large number of studies were conducted to verify the equivalence across cultures of the dimensional Alternative Model of Personality Disorders (DSM-5 Section III). These studies have provided information on configural invariance (i.e., the facets that compose the domains are the same) and metric invariance (i.e., facet-domain relationships are equal across groups), but not on the stricter scalar invariance (i.e., the baseline levels of the facets are the same), which is a prerequisite for meaningfully comparing group means. The present study aims to address this gap. The Personality Inventory for DSM-5 (PID-5) was administered to five samples differing on country and language (Belgium, Catalonia, France, Spain, and Switzerland), with a total of 4,380 participants. Configural and metric invariance were supported, denoting that the model structure was stable across samples. Partial scalar invariance was supported, being minimal the influence of non-invariant facets. This allowed cross-cultural mean comparisons. Results are discussed in light of the sample composition and a possible impact of culture on development of psychopathology.

Effects of several storage media on viability and proliferation capacity of periodontal ligament cells.

AIM: The purpose of this study was to investigate the effect of different storage media on viability and proliferation capacity of periodontal ligament cells. METHODS: Plates with periodontal ligament fibroblasts (PDLF) cells were incubated in skimmed and whole milk, recently prepared Hank's balanced salt solution (HBSS), Save-A-Tooth system's, coconut water, propylene glycol with 20% propolis, egg white, tap water (negative control) at 5 °C and 20 °C, for 24 h. In one of the plates of each temperature, cell viability was determined by MTT assay. In the remaining plates, the wells were filled and incubated with Minimum Essential Medium (MEM) at 37 °C for 24, 48, 72, 96, and 120 h. The proliferation capacity of PDFL cells was also evaluated by MTT assay. Data were statistically analyzed by Kruskal-Wallis, Scheffé and Mann-Whitney tests (α = 5%). RESULTS: At 5 °C, milk maintained more viable cells than other storage media immediately after exposure (0 h) and allowed greater proliferation capacity. At 20 °C, milk and HBSS had similar and allowed similar proliferation ability at 24 and 48 h. From 72 h onwards, capacity to maintain cell viability the proliferation rate of cells incubated in HBSS was superior than milk. At both temperature and experiments, Save-A-Tooth system was similar to tap water. CONCLUSION: Milk and HBSS were more effective in maintaining cellular viability and proliferation capacity than any other storage media. At 5 °C, the most viable alternative was milk. At 20 °C, HBSS had better results.

Trichoderma virens Gl006 and Bacillus velezensis Bs006: a compatible interaction controlling Fusarium wilt of cape gooseberry.

The combination of Trichoderma virens Gl006 and B. velezensis Bs006 as a consortium has high potential to control Fusarium wilt (FW) of cape gooseberry (Physalis peruviana) caused by Fusarium oxysporum f. sp. physali (Foph). However, the interactions between these two microorganisms that influence the biocontrol activity as a consortium have not been studied. Here, we studied the interactions between Gl006 and Bs006 that keep their compatibility under in vitro and greenhouse conditions. Antagonism tests between Gl006 and Bs006 inoculated both individually and in consortium against Foph strain Map5 was carried out on several solid media. The effect of supernatant of each selected microorganism on growth, conidia germination, biofilm formation and antagonistic activity on its partner was also studied. Biocontrol activity by different combinations of cells and supernatants from both microorganisms against Fusarium wilt was evaluated under greenhouse conditions. In vitro antagonism of the consortium against Foph showed a differential response among culture media and showed compatibility among BCA under nutritional conditions close to those of the rhizosphere. The supernatant of Bs006 did not affect the antagonistic activity of Gl006 and vice versa. However, the supernatant of Bs006 promoted the biocontrol activity of Gl006 in a synergistic way under greenhouse, reducing the disease severity by 71%. These results prove the compatibility between T. virens Gl006 and B. velezensis Bs006 as a potential tool to control Fusarium wilt of cape gooseberry.

The effect of paternal exposure to immunosuppressive drugs on sexual function, reproductive hormones, fertility, pregnancy and offspring outcomes: a systematic review.

BACKGROUND: Information regarding the possible influence of immunosuppressive drugs on male sexual function and reproductive outcomes is scarce. Men diagnosed with immune-mediated diseases and a wish to become a father represent an important neglected population since they lack vital information to make balanced decisions about their treatment. OBJECTIVE AND RATIONALE: The aim of this research was to systematically review the literature for the influence of paternal immunosuppressive drug use on many aspects of male sexual health, such as sexual function, fertility, pregnancy outcomes and offspring health outcomes. SEARCH METHODS: A systematic literature search was performed in the bibliographic databases: Embase (via Elsevier embase.com), MEDLINE ALL via Ovid, Cochrane Central Register of Trials (via Wiley) and Web of Science Core Collection. Additionally, Google Scholar and the Clinical trial registries of Europe and the USA were searched. The databases were searched from inception until 31 August 2019. The searches combined keywords regarding male sexual function and fertility, pregnancy outcomes and offspring health with a list of immunosuppressive drugs. Studies were included if they were published in English and if they included original data on male human exposure to immunosuppressive drugs. A meta-analysis was not possible to perform due to the heterogeneity of the data. OUTCOMES: A total of 5867 references were identified, amongst which we identified 161 articles fulfilling the eligibility criteria. Amongst these articles, 50 included pregnancy and offspring outcomes and 130 included sexual health outcomes. Except for large Scandinavian cohorts, most of the identified articles included a small number of participants. While a clear negative effect on sperm quality was evident for sulfasalazine and cyclophosphamide, a dubious effect was identified for colchicine, methotrexate and sirolimus. In three articles, exposure to tumour necrosis factor-α inhibitors in patients diagnosed with ankylosing spondylitis resulted in improved sperm quality. The information regarding pregnancy and offspring outcomes was scant but no large negative effect associated with paternal immunosuppressive drug exposure was reported. WIDER IMPLICATIONS: Evidence regarding the safety of immunosuppressive drugs in men with a wish to become a father is inconclusive. The lack of standardisation on how to evaluate and report male sexual function, fertility and reproduction as study outcomes in men exposed to immunosuppressive drugs is an important contributor to this result. Future research on this topic is needed and should be preferably done using standardised methods.

Sexual function and reproduction can be impaired in men with rheumatic diseases: A systematic review.

BACKGROUND: Information about the possible effect of rheumatic diseases on male sexual function and reproduction (sexual health) is scarce and difficult to summarize. Factors known to impair sexual health, such as inflammation, medication use and hypogonadism can be present in a significant proportion of male patients with rheumatic diseases. OBJECTIVES: The objective of our study was to systematically review the literature for the influence of paternal rheumatic disease on sexual health, such as sexual function, reproductive hormones, male fertility, pregnancy and offspring outcomes. DATA SOURCES: English language articles identified through Embase, MEDLINE, Cochrane Central Register of Controlled Trials (CENTRAL), Web of Science, Google Scholar and the Clinical trial registries of Europe and the USA published until February 2019. STUDY APPRAISAL AND SYNTHESIS METHODS: Literature was synthesized in narrative form and in summary tables. Outcomes were categorized as: sexual function, reproductive hormones, fertility and pregnancy and offspring outcomes. Results are presented per category and per disease. RESULTS: 9735 articles were identified with our search strategy. After removal of duplicates, excluding articles by screening titles and abstracts and assessing eligibility by reading 289 fulltext articles, 87 articles fulfilled the eligibility criteria. All included studies enrolled patients diagnosed with a rheumatic disease and had results at least on one of the outcome categories. Sexual function was the most common category, followed by reproductive hormones, fertility and pregnancy and offspring outcomes. Sexual function is impaired in a high proportion of patients with rheumatic diseases. This was statistically significant in most of the studies where a control group was available. Clinically relevant abnormalities in reproductive hormones were mainly identified in patients with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) and a positive correlation with disease activity were reported. Semen quality in men with rheumatic diseases can be impaired in patients with SLE, SpA, sarcoidosis, BD and MWS. Sperm count and motility were the most common semen quality parameters affected. No negative effect of paternal RA and vasculitis on pregnancy outcomes were reported in 3 studies. No studies reporting the effect of paternal disease on offspring outcomes were identified. LIMITATIONS: Most of the studies included in this review suffer from an inconsistent methodological quality, definitions of outcomes varied in several studies, a wide variety of screening questionnaires and/or diagnostic tools were used and results might only apply to the specific populations that were studied. CONCLUSIONS: This systematic review suggests that sexual health is impaired in men with rheumatic diseases. The degree and extent of sexual health impairment vary per disease. More research is needed to fully understand the link between rheumatic diseases and impaired male sexual health. Meanwhile, rheumatologists should be aware of this association and discuss it with their patients. IMPLICATIONS OF KEY FINDINGS: Sexual health of men with rheumatic diseases can be impaired by the disease itself. Especially in men trying to conceive, information on sexual function, reproductive hormones and sperm quality are needed to identify these problems. Treatment resulting in lower disease activity can improve overall sexual health in man with rheumatic diseases and facilitate their journey to fatherhood. SYSTEMATIC REVIEW REGISTRATION NUMBER: PROSPERO 2018 CRD42018099845.

Human spleen contains different subsets of dendritic cells and regulatory T lymphocytes.

Most knowledge about dendritic cells (DCs) and regulatory T cells in humans has been gathered from circulating cells but little is known about their frequency and distribution in lymphoid organs. This report shows the frequency, phenotype and location of DCs and regulatory T cells in deceased organ donors' spleens. As determined by flow cytometry, conventional/myeloid DCs (cDCs) CD11c(high)HLA-DR(+)CD123(-/low) were 2.3 +/- 0.9% and LIN(-) HLA-DR(+)CD11c(high) 2.1 +/- 0.3% of total spleen cells. Mature CD11c(high)HLA-DR(+)CD83(+) were 1.5 +/- 0.8% and 1.0 +/- 1.6% immature CD11c(high)HLA-DR(+)CD83(-) cDC. There were 0.3 +/- 0.3% plasmacytoid DCs (pDC) CD11c(-/low)HLA-DR(+)CD123(high) and 0.3 +/- 0.1% LIN(-)HLA-DR(+)CD123(high). Cells expressing cDCs markers, BDCA-1 and BDCA-3, and pDCs markers BDCA-2 and BDCA-4 were observed in higher frequencies than DCs with other phenotypes evaluated. CD11c(+), CD123(+) and CD83(+) cells were located in subcapsular zone, T cells areas and B-cell follicles. CD4(+)CD25(high) Tregs were 0.2 +/- 0.2% and CD8(+)CD28(-) comprised 11.5 +/- 8.1% of spleen lymphocytes. FOXP3(+) cells were found in T- and B-cell areas. The improvement in cell separation, manipulation and expansion techniques, will facilitate the manipulation of donor spleen cells as a part of protocols for induction and maintenance of allograft tolerance or treatment of autoimmune diseases.

Oocyte structure and ultrastructure in the Mexican silverside fish Chirostoma humboldtianum (Atheriniforme: Atherinopsidae).

The structural and ultrastructural features of gonads from endemic Mexican fish have received scarce attention. This study describes the histological and ultrastructural characteristics of oocyte from Chirostoma humboldtianum. The ovary is asynchronic, and as such, most phases of oocyte development are found in the same ovary. The complete process of oogenesis was divided in five stages: oogonium and folliculogenesis, primary growth, cortical alveoli and lipid inclusions, vitellogenesis, and maturation. The presence of big filaments, which appear at the end of primary growth, induces some common follicular adaptation. During primary growth, abundant ribosomes, the rough endoplasmic reticulum, and mitochondria are grouped in the cytoplasm. At the end of this stage, the Z1 layer of the chorion is developed, while microvilli start to be evident. In the cortical alveoli and lipid droplets phase, intense PAS positive vesicles, some of them containing nucleoid material, are observed in the peripheral cytoplasm and the lipid droplets take a more central position. In vitellogenesis, the proteic yolk accumulates in a centripetal way while the chorion is completely formed. During maturation, the germinal vesicle migrates to the animal pole, meiosis is restored, and there is nuclear breakdown. The oocyte increases its size and holds some oil droplets and a big fluid mass of yolk. On the outside, filaments completely surround the oocyte.

Nucleologenesis: U3 snRNA-containing prenucleolar bodies move to sites of active pre-rRNA transcription after mitosis.

We have investigated the distribution of U3 snRNA and rRNA in HeLa cells and normal rat kidney cells during interphase and mitosis. U3 snRNA, known to be involved in pre-rRNA processing, was detected in nucleoli and coiled bodies during interphase, whereas rRNA was distributed in the nucleoli and throughout the cytoplasm. By comparison, ribosomal protein S6 was detected in nucleoli, coiled bodies, and in the cytoplasm. During nucleologenesis, pre-rRNA was observed in newly forming nucleoli during late telophase but not in prenucleolar bodies (PNBs), whereas U3 snRNA was detected in forming nucleoli and PNBs. Similar findings to those reported here for the localization of U3 snRNA have been reported previously for the U3 small nuclear ribonucleoprotein fibrillarin. These results suggest that components involved in pre-rRNA processing localize to discrete PNBs at the end of mitosis. The nucleolus is formed at specific telophase domains (nucleolar organizing regions) and the PNBs, containing factors essential for pre-rRNA processing, are recruited to these sites of rRNA transcription and processing.

Additively manufactured MEMS multiplexed coaxial electrospray sources for high-throughput, uniform generation of core-shell microparticles.

This study reports the first MEMS multiplexed coaxial electrospray sources in the literature. Coaxial electrospraying is a microencapsulation technology based on electrohydrodynamic jetting of two immiscible liquids, which allows precise control with low size variation of the geometry of the core-shell particles it generates, which is of great importance in numerous biomedical and engineering applications, e.g., drug delivery and self-healing composites. By implementing monolithic planar arrays of miniaturized coaxial electrospray emitters that work uniformly in parallel, the throughput of the compound microdroplet source is greatly increased, making the microencapsulation technology compatible with low-cost commercial applications. Miniaturized core-shell particle generators with up to 25 coaxial electrospray emitters (25 emitters cm-2) were fabricated via stereolithography, which is an additive manufacturing process that can create complex microfluidic devices at a small fraction of the cost per device and fabrication time associated with silicon-based counterparts. The characterization of devices with the same emitter structure but different array sizes demonstrates uniform array operation. Moreover, the data demonstrate that the per-emitter current is approximately proportional to the square root of the flow rate of the driving liquid, and it is independent of the flow rate of the driven liquid, as predicted by the theory. The core/shell diameters and the size distribution of the generated compound microparticles can be modulated by controlling the flow rates fed to the emitters.