Highly Enantioselective Lewis Acid Catalyzed Conjugate Addition of Imidazo[1,2-a]pyridines to α,ß-Unsaturated 2-Acylimidazoles under Mild Conditions.

A highly enantioselective protocol for the conjugate addition of 2-arylimidazo[1,2-a]pyridines and other imidazo derivatives to α,ß-unsaturated 2-acylimidazoles is described. The method uses a previously reported chiral-at-metal rhodium catalyst and provides the corresponding adducts in yields of 25-98% with enantioselectivities up to er > 99:1. Additionally, the transformation proceeds under mild conditions using ethanol as the solvent at room temperature.

Diffuse large B-cell lymphoma affecting the third eyelid of two young adult FeLV-infected cats.

Lymphoma is one of the most frequently diagnosed tumors in FeLV-infected cats. Extranodal lymphomas include lymphomas in ocular and periocular tissues, such as the third eyelid (TEL), which is an uncommonly diagnosed type of lymphoma in cats. This study aimed to describe the clinicopathological features of lymphoma in the TEL of two FeLV-infected cats. A retrospective study in two anatomic pathology laboratories was performed, and two cases met the inclusion criteria. A 4-year-old, female cat (case 1), positive for FIV gp40 antigen and FeLV gp70 antigen by immunohistochemistry (IHC), and a male cat of 22 months old (case 2), positive both serologically for FeLV p27 antigen and for FeLV gp70 antigen by IHC, were referred to veterinary clinics with unilateral swelling and mass in the TEL, which had rapid growth in case 2. Histological and immunohistochemical analysis established the final diagnosis of diffuse large B-cell lymphoma in both cases. The cat in case 2 died 4 months after the diagnosis, with clinical worsening prior to death. Necropsy was not performed in either case, which precluded the definition of a primary or secondary involvement. However, the ocular lesions were the reason for consultation in both cases and it may have been an initial manifestation of a multicentric disease. Thus, FeLV-infected cats with clinical finding of eyelid swelling or mass formation should have lymphoma as a differential diagnosis, and a complete physical examination should be performed to detect extraocular involvement.

Electrochemically Mediated Atom Transfer Radical Polymerization Driven by Alternating Current.

Alternating current (AC) and pulsed electrolysis are gaining traction in electro(organic) synthesis due to their advantageous characteristics. We employed AC electrolysis in electrochemically mediated Atom Transfer Radical Polymerization (eATRP) to facilitate the regeneration of the activator CuI complex on Cu0 electrodes. Additionally, Cu0 served as a slow supplemental activator and reducing agent (SARA ATRP), enabling the activation of alkyl halides and the regeneration of the CuI activator through a comproportionation reaction. We harnessed the distinct properties of Cu0 dual regeneration, both chemical and electrochemical, by employing sinusoidal, triangular, and square-wave AC electrolysis alongside some of the most active ATRP catalysts available. Compared to linear waveform (DC electrolysis) or SARA ATRP (without electrolysis), pulsed and AC electrolysis facilitated slightly faster and more controlled polymerizations of acrylates. The same AC electrolysis conditions could successfully polymerize eleven different monomers across different mediums, from water to bulk. Moreover, it proved effective across a spectrum of catalyst activity, from low-activity Cu/2,2-bipyridine to highly active Cu complexes with substituted tripodal amine ligands. Chain extension experiments confirmed the high chain-end fidelity of the produced polymers, yielding functional and high molecular-weight block copolymers. SEM analysis indicated the robustness of the Cu0 electrodes, sustaining at least 15 consecutive polymerizations.

Photopharmacological manipulation of amygdala metabotropic glutamate receptor mGlu4 alleviates neuropathic pain.

Neuropathic pain is a common health problem resulting in exacerbated response to noxious and non noxious stimuli, as well as impaired emotional and cognitive responses. Unfortunately, neuropathic pain is also one of the most difficult pain syndromes to manage, highlighting the importance of better understanding the brain regions and neuromodulatory mechanisms involved in its regulation. Among the many interconnected brain areas which process pain, the amygdala is known to play an important role in the integration of sensory and emotional pain signals. Here we questioned the ability of a recently identified neuromodulatory mechanism associated with the metabotropic glutamate receptors mGlu4 in the amygdala to modulate neuropathic pain. In a murine model of peripheral mononeuropathy, we demonstrate that pharmacological activation of amygdala mGlu4 efficiently alleviates sensory and depressive-like symptoms in both male and female mice. Moreover, we reveal a differential modulation of these symptoms. Activating mGlu4 in the contralateral amygdala relative to the side of the mononeuropathy, is necessary and sufficient to relieve both sensory and depressive-like symptoms, while ipsilateral activation solely reduces depressive-like symptoms. Furthermore, using photopharmacology, a recent strategy allowing precise photocontrol of endogenous proteins, we further demonstrate the dynamic alleviation of neuropathic pain through light-dependent facilitation of mGlu4 by a photoswitchable positive allosteric modulator. Finally, coupling photopharmacology and analgesic conditioned place preference, we show a significant pain-reducing effect of mGlu4 activation. Taken together, these data highlight the analgesic potential of enhancing amygdala mGlu4 activity to counteract neuropathy reinforcing its therapeutic interest for the treatment of pathological pain.

Epidemiology of chronic kidney disease in older indigenous peoples of Brazil: findings from a cross-sectional survey.

BACKGROUND: Chronic kidney disease (CKD) is a prevalent disease worldwide, with increasing incidence particularly in low- and middle-income countries. Indigenous communities have poorer CKD outcomes due to limited access to healthcare. They are also experiencing a shift toward a sedentary lifestyle and urbanization-related dietary changes, increasing the risk of CKD-related risk factors. AIM: To determine the prevalence of CKD in older Brazilian indigenous and identify the main associated risk factors. METHODS: This cross-sectional study analyzed demographic and clinical data of 229 older indigenous individuals aged 60 years and above in 2022-2023. CKD was defined as an estimated glomerular filtration rate < 60 mL/min/1.73 m2 or a urinary albumin-creatinine ratio > 30 mg/g. Data were presented categorically and analyzed using the Chi-square test or Fisher's exact test. RESULTS: The prevalence of CKD in the population was 26.6%, with higher prevalence in women and increasing with age. The prevalence of hypertension and diabetes was 67.7% and 24.0%, respectively, and these comorbidities were associated with CKD: hypertension (OR = 5.12; 95% CI 2.2-11.9) and diabetes (OR = 5.5; 95% CI 3.7-8.2). No association was found between the prevalence of CKD and obesity, dyslipidemia, cardiovascular disease, or smoking. DISCUSSION: The study found a higher prevalence of CKD among older indigenous populations in Brazil compared to non-indigenous populations, which is exacerbated by risk factors, such as aging, hypertension, diabetes, and lifestyle changes, emphasizing the importance of early detection and intervention in these communities. CONCLUSION: Older persons' indigenous individuals have a high prevalence of CKD, which is correlated with factors, such as sex, age, diabetes mellitus, and hypertension.

Nanofiltration combined with ozone-based processes for the removal of antineoplastic drugs from wastewater effluents.

Over the past years, there has been an increasing concern about the occurrence of antineoplastic drugs in water bodies. The incomplete removal of these pharmaceuticals from wastewaters has been confirmed by several scientists, making it urgent to find a reliable technique or a combination of techniques capable to produce clean and safe water. In this work, the combination of nanofiltration and ozone (O3)-based processes (NF + O3, NF + O3/H2O2 and NF + O3/H2O2/UVA) was studied aiming to produce clean water from wastewater treatment plant (WWTP) secondary effluents to be safely discharged into water bodies, reused in daily practices such as aquaculture activities or for recharging aquifers used as abstraction sources for drinking water production. Nanofiltration was performed in a pilot-scale unit and O3-based processes in a continuous-flow column. The peroxone process (O3/H2O2) was considered the most promising technology to be coupled to nanofiltration, all the target pharmaceuticals being removed at an extent higher than 98% from WWTP secondary effluents, with a DOC reduction up to 92%. The applicability of the clean water stream for recharging aquifers used as abstraction sources for drinking water production was supported by a risk assessment approach, regarding the final concentrations of the target pharmaceuticals. Moreover, the toxicity of the nanofiltration retentate, a polluted stream generated from the nanofiltration system, was greatly decreased after the application of the peroxone process, which evidences the positive impact on the environment of implementing a NF + O3/H2O2 process.

Neuronal cholinergic signaling constrains norepinephrine activity in the heart.

It is well known that cholinergic hypofunction contributes to cardiac pathology, yet, the mechanisms involved remain unclear. Our previous study has shown that genetically engineered model of cholinergic deficit, the vesicular acetylcholine transporter knockdown homozygous (VAChT KDHOM) mice, exhibit pathological cardiac remodeling and a gradual increase in cardiac mass with aging. Given that an increase in cardiac mass is often caused by adrenergic hyperactivity, we hypothesized that VAChT KDHOM mice might have an increase in cardiac norepinephrine (NE) levels. We thus investigated the temporal changes in NE content in the heart from 3-, 6-, and 12-mo-old VAChT mutants. Interestingly, mice with cholinergic hypofunction showed a gradual elevation in cardiac NE content, which was already increased at 6 mo of age. Consistent with this finding, 6-mo-old VAChT KDHOM mice showed enhanced sympathetic activity and a greater abundance of tyrosine hydroxylase positive sympathetic nerves in the heart. VAChT mutants exhibited an increase in peak calcium transient, and mitochondrial oxidative stress in cardiomyocytes along with enhanced G protein-coupled receptor kinase 5 (GRK5) and nuclear factor of activated T-cells (NFAT) staining in the heart. These are known targets of adrenergic signaling in the cell. Moreover, vagotomized-mice displayed an increase in cardiac NE content confirming the data obtained in VAChT KDHOM mice. Establishing a causal relationship between acetylcholine and NE, VAChT KDHOM mice treated with pyridostigmine, a cholinesterase inhibitor, showed reduced cardiac NE content, rescuing the phenotype. Our findings unveil a yet unrecognized role of cholinergic signaling as a modulator of cardiac NE, providing novel insights into the mechanisms that drive autonomic imbalance.

Randomized trial of a photography-aided behavioural intervention to reduce risk factors for caries and malocclusion in high-risk infants.

OBJECTIVE: To assess the efficacy of a photography-aided behavioural intervention in reducing risk factors for dental caries and malocclusion in high-risk infants. MATERIALS AND METHODS: In this randomized trial conducted in a maternity hospital, 55 mothers of recently born infants at high risk of developing oral diseases were allocated to either the intervention (n = 28) or usual care (n = 27). The intervention arm received the same usual care plus an enhanced, behaviour-oriented, photography-aided, two-stage (0 and +6 months) educational programme addressing nutritional, behavioural, lifestyle and familial factors that affect child's oral health. The primary outcome was the proportion of children classified as being at a 'low risk' of developing dental caries at the age of 12 months using a modified score based on the Caries-risk Assessment Form of the American Academy of Pediatric Dentistry. Secondary outcomes included risk factors for malocclusion, such as duration of exclusive breastfeeding, pacifier use and bottle-feeding and/or sippy cup usage. RESULTS: At 12 months, the proportion of children considered at low risk for dental caries was significantly higher in the intervention group compared to usual care (71% vs 15%, respectively, relative risk = 4.82, 95% confidence interval = 1.89-12.3, p < 0.001). The median duration of exclusive breastfeeding in the intervention group was 1.7 times higher than in the control arm (5 months vs 3 months, p = 0.03). CONCLUSION: Altogether, our findings provide evidence that a low-cost, two-stage preventive strategy using photographs to deliver a stronger visual impact might significantly reduce the incidence of risk factors for dental caries and malocclusion in 12-month-old children.

The Class I HDAC Inhibitor, MS-275, Prevents Oxaliplatin-Induced Chronic Neuropathy and Potentiates Its Antiproliferative Activity in Mice.

Oxaliplatin, the first-line chemotherapeutic agent against colorectal cancer (CRC), induces peripheral neuropathies, which can lead to dose limitation and treatment discontinuation. Downregulation of potassium channels, which involves histone deacetylase (HDAC) activity, has been identified as an important tuner of acute oxaliplatin-induced hypersensitivity. MS-275, a class I histone deacetylase inhibitor (HDACi), prevents acute oxaliplatin-induced peripheral neuropathy (OIPN). Moreover, MS-275 exerts anti-tumor activity in several types of cancers, including CRC. We thus hypothesized that MS-275 could exert both a preventive effect against OIPN and potentially a synergistic effect combined with oxaliplatin against CRC development. We first used RNAseq to assess transcriptional changes occurring in DRG neurons from mice treated by repeated injection of oxaliplatin. Moreover, we assessed the effects of MS-275 on chronic oxaliplatin-induced peripheral neuropathy development in vivo on APCMin/+ mice and on cancer progression when combined with oxaliplatin, both in vivo on APCMin/+ mice and in a mouse model of an orthotopic allograft of the CT26 cell line as well as in vitro in T84 and HT29 human CRC cell lines. We found 741 differentially expressed genes (DEGs) between oxaliplatin- and vehicle-treated animals. While acute OIPN is known as a channelopathy involving HDAC activity, chronic OIPN exerts weak ion channel transcriptional changes and no HDAC expression changes in peripheral neurons from OIPN mice. However, MS-275 prevents the development of sensory neuropathic symptoms induced by repeated oxaliplatin administration in APCMin/+ mice. Moreover, combined with oxaliplatin, MS-275 also exerts synergistic antiproliferative and increased survival effects in CT26-bearing mice. Consistently, combined drug associations exert synergic apoptotic and cell death effects in both T84 and HT29 human CRC cell lines. Our results strongly suggest combining oxaliplatin and MS-275 administration in CRC patients in order to potentiate the antiproliferative action of chemotherapy, while preventing its neurotoxic effect.

Degradation of Neonicotinoids and Caffeine from Surface Water by Photolysis.

Along with rapid social development, the use of insecticides and caffeine-containing products increases, a trend that is also reflected in the composition of surface waters. This study is focused on the phototreatment of a surface water containing three neonicotinoids (imidacloprid, thiamethoxam, and clothianidin) and caffeine. Firstly, the radiation absorption of the target pollutants and the effect of the water matrix components were evaluated. It was observed that the maximum absorption peaks appear at wavelengths ranging from 246 to 274 nm, and that the water matrix did not affect the efficiency of the removal of the target pollutants. It was found that the insecticides were efficiently removed after a very short exposure to UV irradiation, while the addition of hydrogen peroxide was needed for an efficient caffeine depletion. The electrical energy per order was estimated, being the lowest energy required (9.5 kWh m-3 order-1) for the depletion of thiamethoxan by indirect photolysis, and a concentration of hydrogen peroxide of 5 mg dm-3. Finally, a preliminary evaluation on the formation of by-products reveals that these compounds play a key role in the evolution of the ecotoxicity of the samples, and that the application of direct photolysis reduces the concentration of these intermediates.

Attenuation of intestinal inflammation in IL-10 deficient mice by a plasmid carrying Lactococcus lactis strain.

BACKGROUND: Inflammatory bowel diseases (IBD) are intestinal disorders characterized by inflammation in the gastrointestinal tract (GIT) and to date, no efficient treatments exist. Interleukin-10 (IL-10), one of the most important anti-inflammatory cytokines of the immune response, has been under study due to its potential for IBD therapy; however, systemic treatments lead to undesirable side effects and oral administration is limited due to its quick degradation. To avoid these bottlenecks, we previously engineered an invasive Lactococcus lactis (L. lactis) strain capable of delivering, directly to host cells, a eukaryotic DNA expression vector coding for IL-10 of Mus musculus (pValac:il-10) that diminished inflammation in two induced mouse models of intestinal inflammation. Thus, the aim of this study was to analyze its therapeutic effect in the IL-10-deficient mouse model (IL-10-/-) that spontaneously and gradually develops an inflammation that modifies the immune system and resembles Crohn's disease (CD) in humans, and evaluate if it would also diminish and/or prevent the onset of this disease. RESULTS: Oral administration of L. lactis MG1363 FnBPA+ (pValac:il-10) to IL-10-/- mice not only led to IL-10 production by these, but consequently also diminished the severe development of the disease, with animals showing lower macroscopic scores and histological damages, increased IL-10 levels and tendency to lower pro-inflammatory cytokine levels. CONCLUSIONS: The results of this study, together with the previously published ones using this DNA delivery-based strategy, show that it is capable of creating and maintaining an anti-inflammatory environment in the GIT and thus effectively diminish the onset of inflammation in various mouse models.

Determination of the ideal volume of activating solution and the optimal ratio of spermatozoa per oocyte for Prochilodus brevis fertilization.

Prochilodus brevis is a rheophilic species with a threatened natural population that promotes studies aimed at optimizing reproduction in captivity. The correct quantity of inseminating dose and activating solution volume significantly improves fertilization rates, thereby increasing productivity in captivity. The objective of this study was to determine the proportion of sperm per oocyte and the ideal volume of activating solution to be used in the assisted fertilization of P. brevis. Gametes were collected and fertilization performed in two steps. In step 1, the ideal proportion of spermatozoa was determined based on the fertilization rate:oocyte by testing six doses of semen: D1 = 30 × 103, D2 = 150 × 103, D3 = 300 × 103, D4 = 3 × 106, D5 = 5 × 106, and D6 = 10 × 106. In step 2, the fertilization and hatching rates were evaluated using different volumes of activating solution (V1 - 25 ml, V2 - 50 ml, V3 - 75 ml,V4 - 100 ml, V5 - 125 ml, and V6 - 150 ml). A linear regression equation was estimated from steps 1 and 2. The Student-Newman-Keuls test was used to compare the means. In step 1, the percentage of fertilization increased linearly, reaching a plateau of 51.69%. In step 2, the best fertilization rates were obtained with an estimated ideal volume of 75.64 ml per 2 ml of oocytes. Therefore, the proportion of 928,410.29 sperm:oocyte, associated with the volume of 75.64 ml of water per 2 ml of oocytes, provided the maximum reproductive performance for P. brevis.

Analysis of motor performance and histomorphometry of skeletal muscles of rats exercised after cerebral ischemia.

INTRODUCTION: Cerebral ischemia causes muscle atrophy and motor incoordination in animals, impairing motor performance. OBJECTIVE: Thus, the objective of the present study was to analyze the motor performance and histomorphometry of the biceps brachii, soleus, and anterior tibialis muscles of rats submitted to a treadmill training program after induction of cerebral ischemia by the transient occlusion of the middle cerebral artery (MCAO). MATERIALS AND METHODS: To do so, twenty-four 30-day-old Wistar rats were utilized. The exercises were performed for 6 weeks on a leveled treadmill, for 30 min/day, 5 days a week, at a controlled speed of 8 m/min, and the MCAO surgery took place for 60 min. The animals were divided into the following experimental groups: SC (n = 6): control animals that underwent false surgery (operation without the induction of cerebral ischemia) and did not exercise (sedentary); CEA (n = 6): control animals exercised on the treadmill after the false surgery; SI (n = 6): sedentary ischemic animals; IEA (n = 6): animals exercised on the treadmill after the induction of cerebral ischemia. RESULTS: The type-II fibers atrophy in muscle soleo was observed in groups CEA (5125.63 ± 158.83) and SI (4982.38 ± 248.16) when compared to the SC (5927.98 ± 106.17) and IEA (6526.73 ± 195.08) groups. CONCLUSION: It is concluded that the low intensity physical exercise performed on the treadmill for six weeks after the induction of cerebral ischemia by OMCA, promoted a decrease in the atrophy of muscles important of the ischemic animals, benefiting their motor performance.

Changes in DNA Methylation and Gene Expression of Insulin and Obesity-Related Gene PIK3R1 after Roux-en-Y Gastric Bypass.

Weight regulation and the magnitude of weight loss after a Roux-en-Y gastric bypass (RYGB) can be genetically determined. DNA methylation patterns and the expression of some genes can be altered after weight loss interventions, including RYGB. The present study aimed to evaluate how the gene expression and DNA methylation of PIK3R1, an obesity and insulin-related gene, change after RYGB. Blood samples were obtained from 13 women (35.9 ± 9.2 years) with severe obesity before and six months after surgical procedure. Whole blood transcriptome and epigenomic patterns were assessed by microarray-based, genome-wide technologies. A total of 1966 differentially expressed genes were identified in the pre- and postoperative periods of RYGB. From these, we observed that genes involved in obesity and insulin pathways were upregulated after surgery. Then, the PIK3R1 gene was selected for further RT-qPCR analysis and cytosine-guanine nucleotide (CpG) sites methylation evaluation. We observed that the PI3KR1 gene was upregulated, and six DNA methylation CpG sites were differently methylated after bariatric surgery. In conclusion, we found that RYGB upregulates genes involved in obesity and insulin pathways.

Mucosal delivery of Lactococcus lactis carrying an anti-TNF scFv expression vector ameliorates experimental colitis in mice.

BACKGROUND: Anti-Tumor Necrosis Factor-alpha therapy has become clinically important for treating inflammatory bowel disease. However, the use of conventional immunotherapy requires a systemic exposure of patients and collateral side effects. Lactic acid bacteria have been shown to be effective as mucosal delivering system for cytokine and single domain antibodies, and it is amenable to clinical purposes. Therefore, lactic acid bacteria may function as vehicles for delivery of therapeutic antibodies molecules to the gastrointestinal tract restricting the pharmacological effect towards the gut. Here, we use the mucosal delivery of Lactococcus lactis carrying an anti-TNFα scFv expression plasmid on a DSS-induced colitis model in mice. RESULTS: Experimental colitis was induced with DSS administered in drinking water. L. lactis carrying the scFv expression vector was introduced by gavage. After four days of treatment, animals showed a significant improvement in histological score and disease activity index compared to those of untreated animals. Moreover, treated mice display IL-6, IL17A, IL1ß, IL10 and FOXP3 mRNA levels similar to health control mice. Therefore, morphological and molecular markers suggest amelioration of the experimentally induced colitis. CONCLUSION: These results provide evidence for the use of this alternative system for delivering therapeutic biopharmaceuticals in loco for treating inflammatory bowel disease, paving the way for a novel low-cost and site-specific biotechnological route for the treatment of inflammatory disorders.

A novel human pain insensitivity disorder caused by a point mutation in ZFHX2.

Chronic pain is a major global public health issue causing a severe impact on both the quality of life for sufferers and the wider economy. Despite the significant clinical burden, little progress has been made in terms of therapeutic development. A unique approach to identifying new human-validated analgesic drug targets is to study rare families with inherited pain insensitivity. Here we have analysed an otherwise normal family where six affected individuals display a pain insensitive phenotype that is characterized by hyposensitivity to noxious heat and painless bone fractures. This autosomal dominant disorder is found in three generations and is not associated with a peripheral neuropathy. A novel point mutation in ZFHX2, encoding a putative transcription factor expressed in small diameter sensory neurons, was identified by whole exome sequencing that segregates with the pain insensitivity. The mutation is predicted to change an evolutionarily highly conserved arginine residue 1913 to a lysine within a homeodomain. Bacterial artificial chromosome (BAC) transgenic mice bearing the orthologous murine p.R1907K mutation, as well as Zfhx2 null mutant mice, have significant deficits in pain sensitivity. Gene expression analyses in dorsal root ganglia from mutant and wild-type mice show altered expression of genes implicated in peripheral pain mechanisms. The ZFHX2 variant and downstream regulated genes associated with a human pain-insensitive phenotype are therefore potential novel targets for the development of new analgesic drugs.awx326media15680039660001.

Characterization of tomato processing by-product for use as a potential functional food ingredient: nutritional composition, antioxidant activity and bioactive compounds.

Tomato pomace, a by-product generated during tomato processing, was collected at a large tomato processing industry. The by-product was mainly constituted of tomato skin (61.5%), and presented high moisture content (66.58 g.100g-1 wet basis). Among the nutrients, the highest content was of dietary fibre, followed by proteins and fat (50.74, 20.91, 14.14 g.100g-1 d.w., respectively). The pomace has high in vitro antioxidation capacity, especially when measured with the TEAC assay (224.81 µmol Trolox equivalent 100g-1 d.w.). This is due especially to the high amount of lycopene remaining in the by-product after processing (446.9 µg.g-1 d.w). The waste was fractioned into skin and seed fractions by sedimentation, resulting in the increase of lycopene yield by 55%, when using skin fraction as the source material in place of the whole pomace. This by-product shows great potential for being used as a source of the ingredients of high nutritional value, especially dietary fibre and lycopene.

Solvent-Free Process for the Development of Photocatalytic Membranes.

This work described a new sustainable method for the fabrication of ceramic membranes with high photocatalytic activity, through a simple sol-gel route. The photocatalytic surfaces, prepared at low temperature and under solvent-free conditions, exhibited a narrow pore size distribution and homogeneity without cracks. These surfaces have shown a highly efficient and reproducible behavior for the degradation of methylene blue. Given their characterization results, the microfiltration photocatalytic membranes produced in this study using solvent-free conditions are expected to effectively retain microorganisms, such as bacteria and fungi that could then be inactivated by photocatalysis.

Detection of Leishmania infantum DNA in conjunctival swabs of cats by quantitative real-time PCR.

Although some studies have investigated the potential role of cats as a reservoir for Leishmania, their role in the epidemiology of visceral leishmaniasis (VL) is still poorly understood. Molecular diagnostic techniques are an important tool in VL diagnosis, and PCR shows high sensitivity and specificity for Leishmania spp. detection. Quantitative real-time PCR (qPCR) is a method that permits quantitative analysis of a large number of samples, resulting in more sensitive, accurate, and reproducible measurements of specific DNA present in the sample. This study compared real-time PCR (qPCR) and conventional PCR (cPCR) for detection of Leishmania spp. in blood and conjunctival swab (CS) samples of healthy cats from a non-endemic area in the state of São Paulo, Brazil. Of all CS samples, 1.85% (2/108) were positive for Leishmania spp. by both cPCR as qPCR (kappa index = 1), indicating excellent agreement between the two methods. The DNA from the two CS-cPCR- and CS-qPCR-positive samples was further tested with a PCR test amplifying the Leishmania spp. discriminative rRNA internal transcribed spacer 1 (ITS 1), of which one sample generated a 300-350-bp DNA fragment whose size varies according to the Leishmania species. Following sequencing, the fragment showed 100% similarity to a GenBank L. infantum sequence obtained from a cat in Italy. In conclusion, the association of qPCR and CS proved to be effective for detection of Leishmania in cats. Conjunctival swab samples were shown to be a practical and better alternative to blood samples and may be useful in the diagnosis and studies of feline leishmaniasis.

Direct Determination of Nimesulide in Natural Waters and Wastewater by Cathodic Stripping Voltammetry.

Pharmaceuticals are included in a group of compounds considered as contaminants of emerging concern (CECs) in environmental matrices. The quantification of CECs is typically accomplished using chromatographic methods that require several sample pretreatment steps. The current study proposes a voltammetric method to quantify directly nimesulide (NIM) in aqueous samples. The voltammetric parameters were optimized and chosen based on the profile of the voltammograms and peak current intensity. The adequacy of the analytical method was evaluated using validation criteria, such as accuracy, selectivity, linearity, detection, and quantification limits. Linearity was assessed by a standard addition curve at a concentration range of 0.5-130 µg L-1 of NIM. The limit of quantification was 0.50 µg L-1, with 60 s of preconcentration time. Accuracy was expressed as recovery percentages of NIM and ranged from 111.4 to 119.8%. The voltammetric method herein proposed has several advantages over others already used to determine NIM, such as fewer sample pretreatment steps, faster, and cheaper analyses. NIM was detected in wastewater samples at concentration levels ranging from 101.7 to 385.0 µg L-1. This result is the first evidence about NIM occurrences in environmental matrices in the area surrounding the Itaipu Lake reservoir in Brazil.