OPTICAL COHERENCE TOMOGRAPHY ANGIOGRAPHY BIOMARKERS AND MICROPERIMETRY FEATURES IN BEHÇET'S UVEITIS.

PURPOSE: To evaluate the parafoveal macular microvasculature and the macular function in patients with retinal vasculitis associated with Behçet's uveitis. METHODS: In 14 patients with inactive Behçet's uveitis and 26 control individuals (13 with nonocular Behçet's syndrome and 13 healthy subjects), we analyzed the retinal nerve fiber layer, ganglion cell layer, full retinal thickness, foveal avascular zone area and sectorial parafoveal vascular density in the superficial vascular plexus, intermediate capillary plexus, and deep capillary plexus using SPECTRALIS optical coherence tomography (OCT) 2 and OCT angiography. Macular sensitivity was analyzed using an MP-3 microperimeter. RESULTS: Eighteen eyes (78%) had a best-corrected visual acuity ≥ 20/25. Significant differences were found in Behçet's uveitis in comparison with the controls on the OCT and OCT angiography: 14.8%, 22.4%, and 14.9% ganglion cell layer thinning in the global, nasal, and inferior sectors, respectively; 6%, 13.2%, and 7.5% full retinal thickness thinning in the superior, nasal, and inferior sectors; and 16.8%, 14.9%, 23.6%, 15.8%, and 12.6% mean deep capillary plexus density reduction in the global, superior, nasal, inferior, and temporal sectors. Microperimetry data demonstrated significant mean reductions of 21% and 23.6% in central and average macular sensitivities and 28.8%, 40.4%, 27.7%, and 24.2% in the superior, nasal, inferior, and temporal sectors, respectively. Outer plexiform layer elevations were observed in Behçet's uveitis (69.6%). CONCLUSION: Behçet's uveitis presented structural and functional macular damage despite good best-corrected visual acuity, mainly affecting the nasal sector and the deep capillary plexus. On OCT and OCT angiography, quantitative and qualitative changes can be valuable biomarkers of ocular involvement in Behçet's syndrome.

Antibiotic resistance in wastewater treatment plants: understanding the problem and future perspectives.

Antibiotics residues (AR), antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARG) are a new class of water contaminants, due to their adverse effects on aquatic ecosystems and human health. Contamination of water bodies occurs mainly by the excretion of antibiotics incompletely metabolized by humans and animals and is considered the main source of contamination of antibiotics in the environment. Given the imminent threat, the World Health Organization (WHO) has categorized the spread of antibiotics as one of the top three threats to public health in the twenty-first century. The Urban Wastewater Treatment Plants (UWWTP) bring together AR, ARB, ARG, making the understanding of this peculiar environment fundamental for the investigation of technologies aimed at combating the spread of bacterial resistance. Several methodologies have been employed focusing on reducing the ARB and ARG loads of the effluents, however the reactivation of these microorganisms after the treatment is widely reported. This work aims to elucidate the role of UWWTPs in the spread of bacterial resistance, as well as to report the efforts that have been made so far and future perspectives to combat this important global problem.

Self-perception, quality of life and ease of catheterization in patients with continent urinary diversion with the mitrofanoff principle

ABSTRACT Purpose: Continent urinary diversion (CUD) with the Mitrofanoff principle stands as an alternative to urethral catheterization by a route other than the urethra. The aim of the study was to determine self-perception of health-related quality of life (HRQoL), ease of catheterization and global and cosmetic outcomes in patient's dependent on Mitrofanoff catheterization. Materials and methods: Records of all patients who underwent CUD with the Mitrofanoff principle between 2012 to 2018 were reviewed. Data were collected and analysed retrospectively from medical charts. We assessed HRQoL with the EuroQol EQ-5D-3L questionnaire, cosmetic and global satisfaction with a questionnaire designed by the reconstructive urology board and ease of catheterization with a Likert questionnaire adapted from the Intermittent Catheterization Difficulty Questionnaire (ICDQ) validated in patients reliant on retrograde CIC. Results: A total of 25 patients requiring CUD with the Mitrofanoff principle between 2012 and 2018 were assessed, the group was composed mainly of: appendiceal conduits 18 patients (72%) and 7 ileal conduits (Yang-Monti) and three of those requiring Casale (Monti Spiral) and 1 a double Monti technique. Median follow-up was 57 months, median age was 30 years. Visual Analogue Scale (VAS) of the EQ-5D-3L reported a Global health score of 86.5%. Fifty nine percent of the patients had no pain or bleeding with catheterizations. Regarding global satisfaction and cosmetic perception 91% were satisfied with their CUD. Conclusions: CUD is associated with good HRQoL, global satisfaction, ease and painless catheterization, adequate self-perception of cosmetic outcomes and a low complication rate, remaining a safe and viable option.

Self-perception, quality of life and ease of catheterization in patients with continent urinary diversion with the mitrofanoff principle.

PURPOSE: Continent urinary diversion (CUD) with the Mitrofanoff principle stands as an alternative to urethral catheterization by a route other than the urethra. The aim of the study was to determine self-perception of health-related quality of life (HRQoL), ease of catheterization and global and cosmetic outcomes in patient's dependent on Mitrofanoff catheterization. MATERIALS AND METHODS: Records of all patients who underwent CUD with the Mitrofanoff principle between 2012 to 2018 were reviewed. Data were collected and analysed retrospectively from medical charts. We assessed HRQoL with the EuroQol EQ-5D-3L questionnaire, cosmetic and global satisfaction with a questionnaire designed by the reconstructive urology board and ease of catheterization with a Likert questionnaire adapted from the Intermittent Catheterization Difficulty Questionnaire (ICDQ) validated in patients reliant on retrograde CIC. RESULTS: A total of 25 patients requiring CUD with the Mitrofanoff principle between 2012 and 2018 were assessed, the group was composed mainly of: appendiceal conduits 18 patients (72%) and 7 ileal conduits (Yang-Monti) and three of those requiring Casale (Monti Spiral) and 1 a double Monti technique. Median follow-up was 57 months, median age was 30 years. Visual Analogue Scale (VAS) of the EQ-5D-3L reported a Global health score of 86.5%. Fifty nine percent of the patients had no pain or bleeding with catheterizations. Regarding global satisfaction and cosmetic perception 91% were satisfied with their CUD. CONCLUSIONS: CUD is associated with good HRQoL, global satisfaction, ease and painless catheterization, adequate self-perception of cosmetic outcomes and a low complication rate, remaining a safe and viable option.

Removal of microorganisms and antibiotic resistance genes from treated urban wastewater: A comparison between aluminium sulphate and tannin coagulants.

The presence of antibiotic resistant-bacteria (ARB) and antibiotic resistance genes (ARG) in treated effluents of urban wastewater treatment plants (WWTP) may represent a threat to the environment and public health. Therefore, cost-effective technologies contributing to minimize loads of these contaminants in the final effluents of WWTP are required. This study aimed at assessing the capacity of coagulation to reduce the ARB&ARG load in secondary treated urban wastewater (STWW), as well as the impact of the process on the structure and diversity of the bacterial community. Coagulation performance using aluminium sulphate, a synthetic substance, and tannins, a biowaste, was compared. Samples were analysed immediately before (STWW) and after the coagulation treatment (Alu, Tan), as well as after 3-days storage in the dark at room temperature (RSTWW, RAlu, RTan), to assess possible reactivation events. Both coagulants decreased the turbidity and colour and reduced the bacterial load (16S rRNA gene copy number, total heterotrophs (HET), and ARB (faecal coliforms resistant to amoxicillin (FC/AMX) or ciprofloxacin (FC/CIP) up to 1-2 log immediately after the treatment. Both coagulants reduced the load of intl1, but in average, aluminium sulphate was able to decrease the content of the analysed ARGs (blaTEM and qnrS) to lower levels than tannin. Reactivation after storage was observed mainly in RTan. In these samples the load of the culturable populations and qnrS gene prevalence increased, sometimes to values higher than those found in the initial wastewater. Reactivation was also characterized by an increment in Gammaproteobacteria relative abundance in the bacterial community, although with distinct patterns for RTan and RAlu. Curvibacter, Undibacterium and Aquaspirillum were among the most abundant genera in RAlu and Aeromonas, Pseudomonas and Stenotrophomonas in RTan. These bacterial community shifts were in agreement with the variations in the culturable bacterial counts of HET for RTan and FC/CIP for RAlu. In summary, the overall performance of aluminium sulphate was better than that of tannins in the treatment of treated urban wastewater.

Action of sisal (Agave sisalana, Perrine) extract in the in vitro development of sheep and goat gastrointestinal nematodes.

Active compounds from Agave sisalana with antiparasitic action against gastrointestinal nematodes (GINs) could be an alternative to diversify the range of parasite management methods in the livestock sector. The objective of this study was to evaluate the in vitro action of A. sisalana extract on the development of sheep and goat GINs. The extract, obtained from shredded sisal leaves, was utilized at various concentrations in the egg hatch test (EHT), larval development test (LDT), larval feeding inhibition test (LFIT) and adult motility test (AMT). The LC(50) and LC(95) in the EHT were 6.90 and 24.79 mg/mL, in the LDT were 0.041 and 0.067 mg/mL and in the LFIT were 0.053 and 0.24 mg/mL, respectively, showing a dose-dependent relationship. The development and feeding inhibition on L(1) were both 100% at a dose of 0.12 mg/mL. In the AMT there was 100% inhibition at 75 mg/mL after 24h of exposure. The extract of A. sisalana therefore demonstrated significant action on L(1) at 0.12 mg/mL. So, if part of the A. sisalana extract passes through the animal's gastrointestinal system, this material can have a significant effect on the parasites in the feces. This is an interesting approach because it can drastically reduce the pasture contamination as well as the infection of herds.

Systemic leukocyte alterations are associated with invasive uterine cervical cancer.

OBJECTIVE: The aim of the study was to evaluate blood leukocyte counts in patients with uterine cervical neoplasia. METHODS: Patients treated at a university hospital were reviewed retrospectively. Disease progression was monitored, beginning in 1990 to 2002, for at least 5 years. Blood count parameters included absolute leukocyte, neutrophil and lymphocyte counts, leukocytosis (white blood cells > 10³/µL), neutrophilia (neutrophils ≥ 70% of leukocytes), lymphopenia (lymphocytes ≤ 15% of leukocytes), and the neutrophil-lymphocyte ratio (NLR), categorized as less than 5 or 5 or greater. RESULTS: A total of 315 patients were enrolled: 182 (57.8%) with preinvasive neoplasia (cervical intraepithelial neoplasia [CIN] group), 95 (30.1%) with stages I to II (early group), and 38 patients (12.1%) with stages III to IV neoplasia (advanced group). Neutrophil and lymphocyte counts were elevated and reduced, respectively, at advanced stages compared with the CIN group (P < 0.05). Leukocytosis, neutrophilia, lymphopenia, and an NLR of 5 or greater were more frequent at advanced stages compared with the CIN and early-stage groups (P < 0.05). Moreover, neutrophilia was also significantly more frequent at early stage compared with the CIN group. The advanced group with neutrophilia had increased frequency of recidivism and metastasis than patients in the CIN group with neutrophilia (P < 0.05). CONCLUSIONS: Patients with advanced cervical cancer had significantly higher frequency of leukocyte alterations, although they may occur apart from the preinvasive stages. Overall, neutrophilia was the best indicator of cancer invasiveness.

Antimicrobial activity of Artemisia douglasiana and dehydroleucodine against Helicobacter pylori.

ETHNOPHARMACOLOGICAL RELEVANCE: The increasing resistance of Helicobacter pylori to antibiotics demands the search for novel compounds from plant based sources. Artemisia douglasiana Besser is widely used in Cuyo region (Argentina) as folk medicine for the treatment of gastric ailments. AIM OF STUDY: Based on our previous studies that Artemisia douglasiana exert cytoprotective actions against ethanol-induced gastric mucosal injury we assayed the anti-Helicobacter pylori effect of the Artemisia douglasiana extract and its active compound, dehydroleucodine. MATERIALS AND METHODS: The in vitro anti-bacterial activity of Artemisia douglasiana extract and its active compound, dehydroleucodine were determined against one standard strain and six clinical isolates of Helicobacter pylori by using the agar dilution methods. RESULTS: The results showed that both dehydroleucodine and Artemisia douglasiana extract had activity against the microorganism with MICs between 1-8 and 60-120 mg/L, respectively. CONCLUSIONS: Artemisia douglasiana may be a useful alternative treatment strategy principally in eradication of metronidazole and clarithromycin-resistant strain.

Antidiarrheal activity of dehydroleucodine isolated from Artemisia douglasiana.

Dehydroleucodine (DhL), a sesquiterpene lactone obtained from Artemisia douglasiana, was screened for antidiarrheal effects. DhL inhibited castor oil-induced diarrhea in mice by judged by a decrease in the number of wet faeces in the DhL-treatment groups. DhL significantly reduced intestinal transit in mice. Yohimbine and phentolamine counteracted the inhibitory effect of DhL. It is suggested that alpha2-adrenergic receptors mediate the effect of DhL in intestinal motility. DhL reduced also intraluminal accumulation of fluid. Thus, the antidiarrheal activity of DhL is possibly related, at least in part, to its inhibitory action against gastrointestinal motility and the inhibition of enteropooling property.

Comparison between indirect enzyme-linked immunosorbent assays for Anaplasma marginale antibodies with recombinant major surface protein 5 and initial body antigens.

Indirect enzyme-linked immunosorbent assays (ELISAs) based on recombinant major surface protein 5 (rMSP5) and initial body (IB) antigens from a Brazilian isolate of Anaplasma marginale were developed to detect antibodies against this rickettsia in cattle. Both tests showed the same sensitivity (98.2%) and specificities (100% for rMSP5 and 93.8% for IB ELISA) which did not differ statistically. No cross-reactions were detected with Babesia bigemina antibodies, but 5 (rMSP5 ELISA) to 15% (IB ELISA) of cross-reactions were detected with B. bovis antibodies. However, such difference was not statistically significant. Prevalences of seropositive crossbred beef cattle raised extensively in Miranda county, state of Mato Grosso do Sul, Brazil, were 78.1% by rMSP5 ELISA and 79.7% by IB ELISA. In the analysis of sera from dairy calves naturally-infected with A. marginale, the dynamics of antibody production was very similar between both tests, with maternal antibodies reaching the lowest levels at 15-30 days, followed by an increase in the mean optical densities in both ELISAs, suggesting the development of active immunity against A. marginale. Results showed that all calves were seropositive by one-year old, characterizing a situation of enzootic stability. The similar performances of the ELISAs suggest that both tests can be used in epidemiological surveys for detection of antibodies to A. marginale in cattle.

Determination and assay validation of the bioactive sesquiterpene lactone xanthatin isolated from Xanthium cavanillesii using capillary electrophoresis.

This study describes the development and validation of a method for quantification of the antiulcer experimental drug xanthatin in tablets by capillary electrophoresis (CE). Solid oral dosage forms based on xanthatin were designed and assayed on rats. A CE methodology was developed; the parameters evaluated were: background electrolyte composition, concentration and pH, applied voltage and sample preparation. The method was validated in terms of range of linearity, limits of detection (LOD) and quantification (LOQ), accuracy, precision and selectivity and then applied to the pharmaceutical dosage forms. Xanthatin determination was carried out in less than 3 min with a 20 mM sodium tetraborate buffer, pH 9.20. Drug concentration per tablet found was 2.97 +/- 0.2 mg. Calibration plots were linear over at least three orders of magnitude of analyte concentrations, LOD and LOQ were 7.6 and 26 microg mL(-1) respectively. For accuracy evaluation a recovery test was performed, the values being better than 98.6%. With respect to precision, the results obtained were better than 1.02 RSD% (repeatability) and 1.54% (intermediate precision). After the manufacturing process the resulting tablets were biologically active. The methodology developed is useful, simple and rapid for xanthatin determination in tablets.

Relationships in subtribe Diocleinae (Leguminosae; Papilionoideae) inferred from internal transcribed spacer sequences from nuclear ribosomal DNA.

The complete sequences of nuclear ribosomal DNA (nrDNA) internal transcribed spacer regions (ITS/5.8S) were determined for species belonging to six genera from the subtribe Diocleinae as well as for the anomalous genera Calopogonium and Pachyrhizus. Phylogenetic trees constructed by distance matrix, maximum parsimony and maximum likelihood methods showed that Calopogonium and Pachyrhizus were outside the clade Diocleinae (Canavalia, Camptosema, Cratylia, Dioclea, Cymbosema, and Galactia). This finding supports previous morphological, phytochemical, and molecular evidence that Calopogonium and Pachyrhizus do not belong to the subtribe Diocleinae. Within the true Diocleinae clade, the clustering of genera and species were congruent with morphology-based classifications, suggesting that ITS/5.8S sequences can provide enough informative sites to allow resolution below the genus level. This is the first evidence of the phylogeny of subtribe Diocleinae based on nuclear DNA sequences.

Evaluation of diuretic activity of the constituents of Clematis montevidensis Spreng. (Ranunculaceae) in rats.

The effects of Clematis montevidensis Spreng. (Ranunculaceae) on urinary excretion of water, sodium and potassium were investigated in rats loaded with isotonic saline solution. The data reported in the present work indicate that the infusions of the root and aerial part of Clematis montevidensis showed a moderate diuretic activity. This effect could be due, at least in part, to the presence of oleanolic acid isolated from this plant.

Cytoprotective effects of chalcones from Zuccagnia punctata and melatonin on the gastroduodenal tract in rats.

The effects of 2',4'-dihydroxychalcone and 2',4'-dihydroxy-3'-methoxychalcone from Zuccagnia punctata Cav. (Fabaceae) and melatonin administration on ethanol-induced gastroduodenal injury were investigated in rats. Both chalcones showed significant preventive effects in treatment with melatonin previous to the necrotising agent. These effects could be due, in part, to the radical scavenging activity of the melatonin.

Effects of Zuccagnia punctata on the gastrointestinal tract in rats and mice.

A pharmacological evaluation of Zuccagnia punctata Cav. (Fabaceae) on the gastrointestinal tract was made in rats and mice. 2',4'-dihydroxychalcone and 2',4'-dihydroxy-3'-methoxychalcone were isolated from Zuccagnia punctata. The data reported in the present work indicate that the acetone extract and infusion of Zuccagnia punctata reduced intestinal transit in rats and mice and offered protection against ethanol-induced ulceration in rats. The Z. punctata effect could be due, in part, to the presence of 2',4'-dihydroxychalcone and 2',4'-dihydroxy-3'-methoxychalcone in this plant.

Expression and purification of the recombinant ConBr (Canavalia brasiliensis lectin) produced in Escherichia coli cells.

ConBr, a D-glucose/D-mannose-specific lectin from Canavalia brasiliensis seeds, was produced in Escherichia coli from a (c)DNA clone subcloned to pET15b expression vector. The recombinant lectin (rConBr) was purified by one-step immobilized metal-affinity chromatography using an amino-terminal hexahistidine tag. By SDS-PAGE and Western blot, rConBr was highly pure with an apparent molecular mass of 37 kDa. N-terminal sequence analysis revealed a single sequence, confirming the identity of the expressed protein as the pre-pro-ConBr.

Diuretic activity of Fabiana patagonica in rats.

A pharmacological evaluation of the diuretic activity of Fabiana patagonica Speg. (Solanaceae) was carried out in isotonic saline loaded rats. The data reported in the present work indicate that the acetone extract and infusion of the aerial part of Fabiana patagonica showed a moderate diuretic activity. This activity could be due, in part, to the presence of oleanolic acid isolated as the major metabolite of F. patagonica.

Antioxidant activity of Artemisia douglasiana besser extract and dehydroleucodine.

The charge (relative concentration) of antioxidants in aqueous extract and dehydroleucodine (DhL) from Artemisia douglasiana Besser was determined employing a procedure based on the quenching of luminol-enhanced chemiluminescence. Total reactive antioxidant potential (TRAP) and total antioxidant reactivity (TAR) values were determined. The data reported in the present work indicate that the extract of Artemisia douglasiana Besser and DhL showed antioxidant capacity. This activity is more pronounced in the extract, suggesting the presence of several antioxidants in Artemisia douglasiana.

Gastric anti-ulcer activity of several alpha,beta-unsaturated carbonyl compounds in rats.

The gastric cytoprotective activity of several molecules containing an alpha,beta-unsaturated carbonyl system is reported. We attributed this gastroprotective activity to the presence of a non-hindered Michael acceptor in the molecules assayed and suggested that the mechanism of protection would involve, at least in part, a nucleophilic attack of the sulphydryl group of the gastric mucosa to the beta carbon of the Michael acceptors of the compounds assayed.

Effect of dehydroleucodine in experimental colitis in rats and mice.

Dehydroleucodine (DhL), a sesquiterpene lactone (SQL) of the guaianolide type isolated from Artemisia douglasiana Besser, shows a pharmacological cytoprotective effect and significantly prevents the formation of gastric and duodenal lesions induced by various necrotising agents in rodents. The effects of DhL, on two models of experimental colitis were examined. Colitis was produced in male Wistar rats by rectal instillation of 5 and 10% acetic acid, following the methods of Eliakim et al. and Le Duc et al., respectively. In mice colitis was produced by rectal instillation of 0.1 ml of 2,4,6-trinitrobenzene sulphonic acid (5 mg in 50% ethanol) (TNB) as previously described by Chin et al. In this study, the administration of DhL 40 mg kg(-1)(1 h before the induction of colitis) significantly decreased mucosal damage. This effect was consistent in both models. The protection provided by DhL was accompanied by significant decreases in diarrhoea and colon weight; and histologically normal mucosa without ulceration and mucus production were observed. This study shows that both TNB and acetic acid colitis can be pharmacologically controlled by DhL. Our results suggest that the protective activity of DhL in experimental colitis is mediated, at least in part, through the increase of glycoprotein synthesis, anti-inflammatory effect and inhibition of COX-2 induction, and by inhibiting the degranulation of cells containing monoamines.