Effectiveness of photo-ozone therapy against equine Pythium insidiosum.

Cutaneous pythiosis is a life-threatening infectious disease. Low-level laser therapy (LLLT) and ozone (O3) have been used individually in the treatment of infected wounds. The goals of the study were a) to characterize the antimicrobial action of the photo-ozone therapy (LLLT-O3) against equine Pythium insidiosum, and b) to assess the cytotoxic potential of the LLLT-O3 in keratinocytes. Specimens of pathogen were isolated from 10 horses. After culturing, 120 hyphae plugs were distributed among four groups (n=30 hyphae plugs/group): LLLT (laser irradiation for 160 sec;), O3 (exposition to O3 for 15 min;), LLLT-O3 (LLLT and O3 treatments in sequence) and control (untreated plugs). The hyphae growth was measured during the first 14 days post-treatment. Where there was an absence of hyphae growth, the plug was recultured for an additional 7 days. The cytotoxic potential of the treatments against HaCaT keratinocytes was assessed by colorimetric assays. The LLLT-O3 and O3 treatments inactivated, respectively, 92.3% (28/30) and 30% (9/30) of the samples. No growth was detected after 7 days reculture of inactivated hyphae plugs on new media. Hyphae growth was visualized in 100% of the control and LLLT hyphae plugs. The viability of HaCaT cells was not affected by the isolated treatments (LLLT and O3), while the LLLT-O3 showed slight cytotoxic effect (20%) when compared to the control group (P<0.05). Photo-ozone therapy inactivated equine P. insidiosum hyphae with minimal cytotoxicity in skin cells in vitro.

Analysis of microbial communities of ocular prostheses and anophthalmic sockets using 16S rRNA gene sequencing.

Ocular prostheses are liable to host microbial adhesion, which can favor conjunctival inflammation. Knowing the microbiota of the ocular prosthesis and anophthalmic socket is important for predicting infection-related pathogens. This study evaluated the microbiota of the prostheses and anophthalmic sockets of six individuals by 16S rRNA sequencing. The microbial abundance and diversity were analyzed using the Operational Taxonomic Units at the genus level. Forty-seven phyla, capturing 1,258 named genera, were recorded in the sample set. In both sites, the most frequent genera were Fusobacterium, Staphylococcus, Prevotella, and Streptococcus. The microbial abundance was higher for the anophthalmic socket than for the prosthesis. Alpha diversity showed no significant differences in bacterial richness or diversity between sites. Although the microbial abundance in the anophthalmic socket was higher, both sites had similar microbiota with high diversity and low dominance among the genera.

Electrodialytic treatment of secondary mining resources for raw materials extraction: Reactor design assessment.

The sustainability of mining activities is compromised due to the high amounts of mining residues generated that have to be disposed of, often in open dams, that may cause environmental deterioration, e.g. release of toxic elements to water supplies. These residues are, however, secondary resources of raw materials. In the case of Panasqueira mine, they even are a source of tungsten, considered a critical raw material. The present work aims to assess the electrodialytic process efficiency for raw materials extraction from Panasqueira mine residues. Experiments were performed with 2 and 3-compartment electrodialytic reactors, applying current intensities between 50 and 100 mA, from 4 to 14 days, and sample suspensions enhanced with NaCl or effluent. Additionally, control experiments with no current application were carried out. The results showed that a 3-compartment reactor operating at 100 mA, with NaCl as supporting electrolyte, presented the highest extraction of copper (13%), tin (10%), tungsten (13%) and arsenic (63%).

Electrodialytic removal of tungsten and arsenic from secondary mine resources - Deep eutectic solvents enhancement.

Tungsten is a critical raw material for European and U.S. economies. Tungsten mine residues, usually considered an environmental burden due to e.g. arsenic content, are also secondary tungsten resources. The electrodialytic (ED) process and deep eutectic solvents (DES) have been successfully and independently applied for the extraction of metals from different complex environmental matrices. In this study a proof of concept demonstrates that coupling DES in a two-compartment ED set-up enhances the removal and separation of arsenic and tungsten from Panasqueira mine secondary resources. Choline chloride with malonic acid (1:2), and choline chloride with oxalic acid (1:1) were the DES that in batch extracted the average maximum contents of arsenic (16%) and tungsten (9%) from the residues. However, when ED was operated at a current intensity of 100 mA for 4 days, the extraction yields increased 22% for arsenic and 11% for tungsten, comparing to the tests with no current. From the total arsenic and tungsten extracted, 82% and 77% respectively were successfully removed from the matrix compartment, as they electromigrated to the anolyte compartment, from where these elements can be further separated. This achievement potentiates circular economy, as the final treated residue could be incorporated in construction materials production, mitigating current environmental problems in both mining and construction sectors.

Quantification and phenotypic characterization of peripheral blood Vδ1 + T cells in chronic lymphocytic leukemia and monoclonal B cell lymphocytosis.

BACKGROUND: Vδ1+ T cells, a subset of γδ T cells, are responsible for innate-like immune responses. Recently, an anti-tumor function mediated by MHC-unrestricted recognition of lipid and stress molecules, has also been described in these cells. This study aimed to quantify and phenotypically characterize circulating Vδ1+ T cells in B cell Chronic Lymphocytic Leukemia (CLL) and Monoclonal B cell lymphocytosis (MBL). METHODS: This study enrolled 58 individuals distributed in five groups: Binet B and C CLL (n = 9), Binet A CLL (n = 26), High count-MBL (n = 10), Low count-MBL (n = 5), and a control group (n = 8). The phenotypic characterization of Vδ1+ T cells, as well as the other T cell subpopulations (CD4+ , CD8+ , CD4+ /CD8+ , and Vδ1- γδT cells), were assessed by flow cytometry, evaluating the frequency of each subset expressing CD27, CD69, and cytoplasmic granzyme B. RESULTS: We observed an increasing percentage of Vδ1+ T cells belonging to CD27- compartment from controls to advanced stages of the disease, which was accompanied by an increasing percentage of these cells expressing granzyme B, a phenotypic pattern that was also observed in the other T cell subpopulations under study since earlier stages of the disease. Moreover, a striking expansion of Vδ1+ T cells in Binet B and C CLL was observed. CONCLUSIONS: These experiment findings point to an expansion of CD27- Vδ1+ T cells with a cytotoxic profile, from controls to advanced stages of the disease, which points to a role of Vδ1+ T cells in the host's anti-tumor responses against clonal B-cells in MBL and CLL. © 2018 Clinical Cytometry Society.

Sustainability of construction materials: Electrodialytic technology as a tool for mortars production.

The reduction of tap water consumption in all activity sectors, including the building industry, is crucial to the sustainability of water resources. Effluents from wastewater treatment plants have the potential to replace freshwater in the construction sector but they contain a critical mixture of impurities, which hampers their use in mortars production. In this work, the viability of using effluent as an alternative to potable water for the production of mortars, after electrodialytic treatment, was assessed. Electrodialytic technology (ED-T) is a proven technique for decontamination of porous and aqueous matrices. ED-T experiments were conducted with 500 mL of effluent for 6, 12 and 24 h, with a current intensity of 25 mA. The results showed that after ED-T 6 h, the removal efficiencies of critical components were above 85% of their initial concentrations. Mortar properties such as setting time, workability, flexural strength, compressive strength and morphology were obtained for 100% effluent and tap water/effluent mixtures (50:50) with and without ED-T pre-treatment. The mortars with the ED-T treated effluent showed similar initial setting times and workability, higher flexural and compressive strength compared to the mortars reference.

Mini-Implants for Mandibular Overdentures: Cost-Effectiveness Analysis alongside a Randomized Trial.

Mandibular overdentures retained by 2 conventional implants have been considered the standard of care for complete edentulism, according to the McGill and York consensuses. However, many patients refuse this treatment modality due to the associated costs and postsurgical discomfort. Mini-implants have the chance to overcome these limitations due to their potentially lower costs and a relatively uncomplicated surgical technique. This study compared treatment costs and incremental cost-effectiveness following the insertion of mini-implants (2 or 4) or 2 standard-size implants for the retention of mandibular overdentures, by means of a randomized clinical trial. In total, 120 edentulous participants (mean age 59.5 ± 8.5 y) were randomly allocated into 3 groups according to treatment received: 4 mini-implants (group 1), 2 mini-implants (group 2), or 2 standard implants (group 3). Treatment costs and outcomes (Oral Health Impact Profile for Edentulous [OHIP-EDENT] and satisfaction with the dentures) were evaluated after 6 mo. Incremental cost-effectiveness ratios (ICERs) were calculated for each intervention in terms of cost per 1-point change in patient outcomes. A 1-way sensitivity analysis was performed considering a 95% confidence interval variation in cost and outcome parameters, represented in tornado diagrams. Overall treatment cost was the lowest for group 2 (average cost: US$318.08), followed by group 1 (US$510.75) and group 3 (US$566.13). Groups did not differ in terms of the length of unscheduled appointments and time spent by participants. In summary, our findings indicate that mandibular overdentures retained by 2 or 4 mini-implants are less costly compared to 2-implant overdentures. Despite the lower costs of overdentures retained by 2 mini-implants, those retained by 4 mini-implants showed further improvement in patient-reported outcomes and reduced costs compared to standard implants ( ClinicalTrials.gov NCT01411683). Knowledge Transfer Statement: This report shows that mini-implant retained overdentures are less costly than overdenture treatment on 2 standard-sized implants. Treatment with 2 mini-implants is an effective procedure to substantially save resources, whereas treatment with 4 mini-implants provides better results from a patient perspective combined with slightly reduced costs compared to the treatment with 2 standard implants. Therefore, mini-implant overdentures may be effective and more accessible than overdentures on 2 standard-size implants for those with limited incomes.

Pro-inflammatory cytokines, IL-1ß and TNF-α, produce persistent compromise in tonic immobility defensive behaviour in endotoxemia guinea-pigs.

AIM: Sepsis has been associated with acute behavioural changes in humans and rodents, which consists of a motivational state and an adaptive response that improve survival. However, the involvement of peripheral cytokines synthesized during systemic inflammation as modulators of the tonic immobility (TI) defensive behaviour remains a literature gap. Our purposes were to characterize the TI defensive behaviour in endotoxemia guinea-pigs at acute phase and after recovery from the initial inflammatory challenge. Furthermore, we investigated whether peri-aqueductal grey matter (PAG) exists as a brain structure related to this behaviour and also pro-inflammatory cytokines, tumour necrosis factor (TNF)-α and interleukin (IL)-1ß, act at this mesencephalic nucleus. METHODS: Endotoxemia was induced by lipopolysaccharide (LPS) administration in guinea-pigs. The parameters evaluated included TI defensive behaviour, survival, cytokines production, as well as neuronal activation and apoptosis in the PAG. RESULTS: Endotoxemia guinea-pigs exhibited a reduction in the duration of TI episodes, starting at 2 h after LPS administration and persisting throughout the experimental period evaluated over 7 days. Moreover, endotoxemia increased the c-FOS immunoreactivity of neurones in the ventrolateral PAG (vlPAG), as well as the caspase-3 expression. The LPS microinjection into vlPAG reproduces the same compromise, that is a decrease in the duration of TI defensive behaviour, observed after the peripheral administration. The immunoneutralization against IL-1ß and TNF-α into vlPAG reverts all the effects produced by peripheral LPS administration. CONCLUSION: Our findings confirm that vlPAG is an important brain structure involved in the behavioural alterations induced by endotoxemia, possibly changing the neuronal activity caused by pro-inflammatory cytokines produced peripherally.

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.

Mini vs. Standard Implants for Mandibular Overdentures: A Randomized Trial.

A mandibular implant-retained overdenture is considered a first-choice treatment for edentulism. However, some aspects limit the use of standard implants-for example, the width of edentulous ridges, chronic diseases, fear, or costs. This randomized trial compared mandibular overdentures retained by 2 or 4 mini-implants with standard implants, considering oral health-related quality of life (OHRQoL), patient satisfaction, and complications such as lost implant. In sum, 120 edentulous men and women (mean age, 59.5 ± 8.5 y) randomly received 4 mini-implants, 2 mini-implants, or 2 standard implants. Participants provided data regarding OHRQoL and satisfaction until 12 mo. Clinical parameters, including implant survival rate, were also recorded. Both 2 and 4 mini-implants led to better OHRQoL, compared with 2 standard implants. Treatment with 4 mini-implants was more satisfying than 2 standard implants, with 2 mini-implants presenting intermediate results. Implant survival rate was 89%, 82%, and 99% for 4 mini-implants, 2 mini-implants, or 2 standard implants, respectively. Overdentures retained by 4 or 2 mini-implants can achieve OHRQoL and satisfaction at least comparable with that of 2 standard implants. However, the survival rate of mini implants is not as high as that of standard implants (ClinicalTrials.gov NCT01411683).

Oxidative stress and mitochondrial dysfunction play a role in myelodysplastic syndrome development, diagnosis, and prognosis: A pilot study.

The imbalance between reactive oxygen species (ROS) production and their elimination by antioxidants leads to oxidative stress. Depending on their concentration, ROS can trigger apoptosis or stimulate cell proliferation. We hypothesized that oxidative stress and mitochondrial dysfunction may participate not only in apoptosis detected in some myelodysplastic syndrome (MDS) patients, but also in increasing proliferation in other patients. We investigated the involvement of oxidative stress and mitochondrial dysfunction in MDS pathogenesis, as well as assessed their diagnostic and prognostic values. Intracellular peroxides, superoxide, superoxide/peroxides ratio, reduced glutathione (GSH), and mitochondrial membrane potential (Δψ(mit)) levels were analyzed in bone marrow cells from 27 MDS patients and 12 controls, by flow cytometry. We observed that all bone marrow cell types from MDS patients had increased intracellular peroxide levels and decreased GSH content, compared with control cells. Moreover, oxidative stress levels were MDS subtype- and risk group-dependent. Low-risk patients had the highest ROS levels, which can be related with their high apoptosis; and intermediate-2-risk patients had high Δψ(mit) that may be associated with their proliferative potential. GSH levels were negatively correlated with transfusion dependency, and peroxide levels were positively correlated with serum ferritin level. GSH content proved to be an accurate parameter to discriminate patients from controls. Finally, patients with high ROS or low GSH levels, as well as high superoxide/peroxides ratio had lower overall survival. Our results suggest that oxidative stress and mitochondrial dysfunction are involved in MDS development, and that oxidative stress parameters may constitute novel diagnosis and/or prognosis biomarkers for MDS.

Short-term post-operative pain and discomfort following insertion of mini-implants for retaining mandibular overdentures: a randomized controlled trial.

The retention of removable dentures by mini-implants is a relatively recent treatment modality and may lead to minimal post-operative trauma. This study compared post-operative pain and discomfort following the insertion of mini-implants (two or four) or two standard-size implants for the retention of mandibular overdentures. One hundred and twenty edentulous participants (mean age 59.5 ± 8.5 years) were randomly allocated into three groups according to received treatment: (GI) four mini-implants, (GII) two mini-implants or (GIII) two standard implants. Seven days after implant insertion, patients answered questions (100-mm VAS) relating to pain, swelling, and discomfort with chewing, speech and hygiene, considering their experiences during the 1st and 6th day. Groups were compared by two-way anova (α = 0.05). All participants (GI: 38; GII: 42; GIII: 40) were analysed after 7 days. At the 6th day, GI felt significantly higher pain than GII and GIII. GI also reported more difficulty in performing oral hygiene practices than GIII during the 1st day. There was no significant difference between groups for the other questions and periods. No participant suffered unexpected side effects. The use of four mini-implants induces more intense post-operative pain at the 6th day than the insertion of two mini- or conventional fixtures, as well as more difficult oral hygiene on the 1st day. Clinicaltrials.gov, NCT01411683; FAPESP, 2011/00688-7 and 2011/23347-0.

Kisspeptin regulates tuberoinfundibular dopaminergic neurones and prolactin secretion in an oestradiol-dependent manner in male and female rats.

Prolactin (PRL) secretion is inhibited by hypothalamic dopamine. Kisspeptin controls luteinising hormone (LH) secretion and is also involved in PRL regulation. We further investigated the effect of kisspeptin-10 (Kp-10) on the activity of tuberoinfundibular dopaminergic (TIDA) neurones and the role of oestradiol (E2 ) in this mechanism. Female and male rats were injected with i.c.v. Kp-10 and evaluated for PRL release and the activity of dopamine terminals in the median eminence (ME) and neurointermediate lobe of the pituitary (NIL). Kp-10 at the doses of 0.6 and 3 nmol increased plasma PRL and decreased 4-dihydroxyphenylacetic acid (DOPAC) levels in the ME and NIL of ovariectomised (OVX), E2 -treated rats but had no effect in OVX. In gonad-intact males, 3 nmol Kp-10 increased PRL secretion and decreased DOPAC levels in the ME but not in the NIL. Castrated males treated with either testosterone or E2 also displayed increased PRL secretion and reduced ME DOPAC in response to Kp-10, whereas castrated rats receiving oil or dihydrotestosterone were unresponsive. By contrast, the LH response to Kp-10 was not E2 -dependent in either females or males. Additionally, immunohistochemical double-labelling demonstrated that TIDA neurones of male rats contain oestrogen receptor (ER)-α, with a higher proportion of neurones expressing ERα than in dioestrous females. The dopaminergic neurones of periventricular hypothalamic nucleus displayed much lower ERα expression. Thus, TIDA neurones express ERα in male and female rats, and kisspeptin increases PRL secretion through inhibition of TIDA neurones in an E2 -dependent manner in both sexes. These findings provide new evidence about the role of kisspeptin in the regulation of dopamine and PRL.

Beyond the limits of oxygen: effects of hypoxia in a hormone-independent prostate cancer cell line.

Prostate cancer (PCa) has a high incidence worldwide. One of the major causes of PCa resistance is intratumoral hypoxia. In solid tumors, hypoxia is strongly associated with malignant progression and resistance to therapy, which is an indicator of poor prognosis. The antiproliferative effect and induced death caused by doxorubicin, epirubicin, cisplatin, and flutamide in a hormone-independent PCa cell line will be evaluated. The hypoxia effect on drug resistance to these drugs, as well as cell proliferation and migration, will be also analyzed. All drugs induced an antiproliferative effect and also cell death in the cell line under study. Hypoxia made the cells more resistant to all drugs. Moreover, our results reveal that long time cell exposure to hypoxia decreases cellular proliferation and migration. Hypoxia can influence cellular resistance, proliferation, and migration. This study shows that hypoxia may be a key factor in the regulation of PCa.

A randomised trial of a simplified method for complete denture fabrication: patient perception and quality.

Complete denture fabrication involves a series of complex technical procedures. Nevertheless, simplified methods may be as effective as conventional ones albeit the lesser use of time and resources, without disadvantage for the patient. This study compared a simplified method for complete denture fabrication to a conventional protocol in terms of oral health-related quality of life (OHRQoL), patient satisfaction and denture quality. Forty-two edentulous patients requesting treatment with complete dentures were randomly allocated into two study groups. Group S received dentures fabricated by a simplified method and Group C received conventionally fabricated dentures. Before interventions and after three and 6 months following insertion, OHRQoL and patient satisfaction were analysed by specific instruments. A prosthodontist assessed denture quality 3 months after delivery. Groups presented no difference for OHRQoL, denture quality and general satisfaction. Differences regarding patient satisfaction with some aspects of the dentures were found after 3 months (S > C), but were insignificant at 6 months. It was concluded that the simplified method is able to produce dentures of a quality comparable to those produced by the conventional method, influencing OHRQoL and patient satisfaction similarly.

A randomised trial on simplified and conventional methods for complete denture fabrication: masticatory performance and ability.

OBJECTIVES: To compare a simplified method to a conventional protocol for complete denture fabrication regarding masticatory performance and ability. METHODS: A sample was formed by edentulous patients requesting treatment with maxillary and mandibular complete dentures. Participants were randomly divided into two groups: Group S, which received dentures fabricated by a simplified method, and Group C (n=21 each), which received conventionally fabricated dentures. After three months following insertion, masticatory performance was evaluated by a colorimetric assay based on chewing two capsules as test food during twenty and forty cycles. Masticatory ability was assessed by a questionnaire with binary answers and a single question answered by means of a 0-10 scale. A third group (DN) formed by seventeen dentate volunteers served as an external comparator. Groups were compared by statistical tests suitable for data distribution (α=0.05). RESULTS: Thirty-nine participants were assessed for three months (twenty from Group C and nineteen from Group S). Groups C and S presented similar masticatory performance which corresponded to approximately 30% of Group DN. Results for masticatory ability showed similarity between S and C, regardless of the assessment method, although an isolate questionnaire item showed more favourable results for the first group. CONCLUSIONS: The simplified method for complete denture fabrication is able to restore masticatory function to a level comparable to a conventional protocol, both physiologically and according to patient's perceptions. CLINICAL SIGNIFICANCE: Although masticatory function is impaired by the loss of natural teeth and dentures can restore only a fraction of such function, patients can benefit from a simplified protocol for complete denture fabrication to the same extent they would by conventional techniques.