Impact of photobiomodulation for oral mucositis on body weight and BMI of patients with head and neck cancer.

PURPOSE: To evaluate the impact of photobiomodulation for oral mucositis on body weight and body mass index (BMI) in head and neck cancer patients. METHODS: Patients were separated into two groups: photobiomodulation (PBM) (n = 17) and SHAM (n = 19). Participants assigned to the PBM group received the photobiomodulation therapy on alternate days at 33 different sites of the oral mucosa (1 J for 10 s per application point) using a low-power aluminum gallium arsenide (AsGaAl) laser device with a maximum output power of 86.7 mW and a wavelength of 660 nm (red). The same protocol was performed in patients allocated in the SHAM group but with the device turned off. At the first and twenty-fourth radiotherapy sessions, participants from both groups underwent an intraoral examination to assess the presence or absence of mucositis using the WHO scale, and measurements of weight and height for BMI calculation were undertaken. RESULTS: All patients had significant differences in weight and BMI throughout the study period. Patients in the PBM group lost less weight (p < 0.01) and had a reduced BMI loss (p < 0.01) in comparison to patients in the SHAM group. Although PBM did not prevent oral mucositis, it decreased its severity (p < 0.01). CONCLUSION: Photobiomodulation for oral mucositis reduced weight loss and prevented a reduction in BMI in patients who underwent chemoradiotherapy for head and neck cancer. Retrospectively registered at https://data.mendeley.com//datasets/4kd7s49wk4/1 .

Real-world data on neoadjuvant endocrine therapy in ER-positive/HER2-negative breast cancer.

PURPOSE: Neoadjuvant endocrine therapy (NET) has been shown to be effective in ER-positive/HER2-negative breast cancer in clinical trials. However, adoption in clinical practice is still limited. Real-world data may provide useful insights into effectiveness, toxicities and quality of care, potentially rendering clinical trial results to the real-world setting. Our purpose was to report real-world data of a cohort of postmenopausal patients submitted to NET. METHODS: This prospective cohort study evaluated 146 postmenopausal female patients with ER-positive/HER2-negative breast cancer treated with NET at three tertiary hospitals between 2016 and 2018. Clinicopathological information were collected prospectively. Preoperative Endocrine Prognostic Index (PEPI) score was calculated for tumors submitted to at least 16 weeks of NET. RESULTS: Median age was 67 years old, and 87.8% had stage I-II disease. Most tumors had histological grade II (76.1%). Median pretreatment Ki67 expression was 10%. Aromatase inhibitor was used in 99.5% of patients, and median treatment duration was 21.0 weeks. No tumor progressed during NET. Breast-conserving surgery was performed in the majority of patients (63.0%), as well as sentinel lymph-node biopsy (76.7%). Pathological complete response rate was 1.0%. 43 patients (29.5%) had PEPI score 0, and 26% had PEPI scores 4-5. Posttreatment Ki67 median expression was 3.0%, and only five tumors (3.4%) showed marked increase in Ki67 expression during treatment. Seven patients (4.8%) had HER2-positive residual disease, and were treated with adjuvant chemotherapy plus trastuzumab. CONCLUSIONS: Our real-world data shows that NET is effective and safe in postmenopausal patients with ER-positive/HER2-negative breast cancer. Postmenopausal status and low-risk luminal tumor features (luminal A-like) should be used as selection criteria to ensure the best results with NET.

Maternal Exposure to Ethanol During Pregnancy and Lactation Affects Glutamatergic System and Induces Oxidative Stress in Offspring Hippocampus.

BACKGROUND: Alcohol abuse during pregnancy leads to intellectual disability and morphological defects in the offspring. The aim of this study was to determine the effect of chronic maternal ethanol (EtOH) consumption during pregnancy and lactation on glutamatergic transmission regulation, energy deficit, and oxidative stress in the hippocampus of the offspring. METHODS: EtOH was administered to dams in drinking water at increasing doses (2 to 20%) from the gestation day 5 to lactation day 21. EtOH and tap water intake by treated and control groups, respectively, were measured daily. RESULTS: Results showed that EtOH exposure does not affect fluid intake over the course of pregnancy and lactation. The toxicity of maternal exposure to EtOH was demonstrated by decreased offspring body weight at experimental age, on postnatal day 21. Moreover, maternal EtOH exposure decreased (45) Ca(2+) influx in the offspring's hippocampus. Corroborating this finding, EtOH increased both Na(+) -dependent and Na(+) -independent glial [(14) C]-glutamate uptake in hippocampus of immature rats. Also, maternal EtOH exposure decreased glutamine synthetase activity and induced aspartate aminotransferase enzymatic activity, suggesting that in EtOH-exposed offspring hippocampus, glutamate is preferentially used as a fuel in tricarboxylic acid cycle instead of being converted into glutamine. In addition, EtOH exposure decreased [U-14C]-2-deoxy-D-glucose uptake in offspring hippocampus. CONCLUSIONS: The decline in glucose transport coincided with increased lactate dehydrogenase activity, suggesting an adaptative response in EtOH-exposed offspring hippocampus, using lactate as an alternative fuel. These events were associated with oxidative damage, as demonstrated by changes in the enzymatic antioxidant defense system and lipid peroxidation. Taken together, the results demonstrate that maternal exposure to EtOH during pregnancy and lactation impairs glutamatergic transmission, as well as inducing oxidative stress and energy deficit in immature rat hippocampus.

Diagnosis and Prevalence of Prominent Lobules in Otoplasty: Analysis of 120 Patients with Prominent Ears.

BACKGROUND: The prominence of the ear lobule is considered an anomaly of secondary importance and receives correspondingly less attention in literature. We reviewed a case series of otoplasty patients and analyzed the prevalence of lobule prominence. METHODS: Records of otoplasties between 2007 and 2013 were reviewed. Inclusion criteria were: (1) primary otoplasties; (2) prominence of both ears; and (3) otoplasties conducted by the main author. Patients were divided into two groups: (1) A general group (GG) containing all the patients, and (2) a lobule correction group (LG)-a subgroup of GG containing only the patients who needed lobule correction. RESULTS: From a total of 291 patients, 120 patients were included in GG and 27 patients in LG. There was no statistical difference between groups GG and LG in terms of age and gender. Preoperative diagnosis of lobule prominence was correctly established in 14 patients; 13 patients were diagnosed during surgery. CONCLUSIONS: Lobule prominence should not be underestimated since its diagnosis may be missed in the preoperative period-nearly 50 % in our case. Also its prevalence (22.5 %) is not as infrequent as some may think. Knowledge of lobule correction techniques before surgery is important to avoid less than optimal results in otoplasty. LEVEL OF EVIDENCE V: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.

Erythrocyte alterations in specimens of Danio rerio caused by exposure to metformin.

The antidiabetic drug metformin is widely prescribed around the world. However, its permanence in different environmental concentrations has been associated with adverse toxicological effects in organisms that do not target its therapeutic action. In the aquatic environment, fish such as the Zebrafish (Danio rerio) have been considered potential bioindicators of environmental impacts and used as experimental models in toxicological studies due to the sensitivity of these species to different types of contaminants, including pharmaceuticals. Thus, this study aimed to analyze metformin's cytotoxic effects on Danio rerio erythrocytes. The animals were submitted to different concentrations of the drug (50 µg/L, 100 µg/L, 150 µg/L, and 10000 µg/L) for 365 days and subsequently observed employing light microscopy and scanning electron microscopy (SEM) to evaluate the alterations that occurred. Exposure of animals to metformin led to significant erythrocyte cell abnormalities across all tested concentrations, with a particularly pronounced effect at the higher concentration previously defined as the NOEC (No Observed Effect Concentration). Remarkable abnormalities included cytoplasmic vacuoles, echinocytes, and vesicle-like cytoplasmic fragments. These findings suggest that metformin, even at concentrations similar to those found in nature and at the NOEC level, exhibits cytotoxic potential in D. rerio, raising concerns about its potential health impacts.

Complete genome sequences and characteristics of mycobacteriophages Diminimus, Dulcita, Glaske16, and Koreni.

Complete genome sequences of four novel mycobacteriophages, Diminimus, Dulcita, Glaske16, and Koreni, isolated from soil are presented. All these bacteriophages belong to subcluster M1, except Koreni that belongs to subcluster A4. Moreover, all have siphovirus morphologies, with genome sizes ranging from 51,055 to 81,156 bp.

Evaluation of Subtle Auditory Impairments with Multiple Audiological Assessments in Normal Hearing Workers Exposed to Occupational Noise.

Recent studies involving guinea pigs have shown that noise can damage the synapses between the inner hair cells and spiral ganglion neurons, even with normal hearing thresholds-which makes it important to investigate this kind of impairment in humans. The aim was to investigate, with multiple audiological assessments, the auditory function of normal hearing workers exposed to occupational noise. Altogether, 60 workers were assessed (30 in the noise-exposure group [NEG], who were exposed to occupational noise, and 30 in the control group [CG], who were not exposed to occupational noise); the workers were matched according to age. The following procedures were used: complete audiological assessment; speech recognition threshold in noise (SRTN); speech in noise (SN) in an acoustic field; gaps-in-noise (GIN); transient evoked otoacoustic emissions (TEOAE) and inhibitory effect of the efferent auditory pathway; auditory brainstem response (ABR); and long-latency auditory evoked potentials (LLAEP). No significant difference was found between the groups in SRTN. In SN, the NEG performed worse than the CG in signal-to-noise ratio (SNR) 0 (p-value 0.023). In GIN, the NEG had a significantly lower percentage of correct answers (p-value 0.042). In TEOAE, the NEG had smaller amplitude values bilaterally (RE p-value 0.048; LE p-value 0.045) and a smaller inhibitory effect of the efferent pathway (p-value 0.009). In ABR, the NEG had greater latencies of wave V (p-value 0.017) and interpeak intervals III-V and I-V in the LE (respective p-values: 0.005 and 0.04). In LLAEP, the NEG had a smaller P3 amplitude bilaterally (RE p-value 0.001; LE p-value 0.002). The NEG performed worse than the CG in most of the assessments, suggesting that the auditory function in individuals exposed to occupational noise is impaired, even with normal audiometric thresholds.

Cerebrospinal fluid fistula as a complication of reverse transcriptase-polymerase chain reaction collection for the detection of coronavirus disease 2019: illustrative cases.

BACKGROUND: The most used method to detect coronavirus disease 2019 during the pandemic is reverse transcriptase-polymerase chain reaction with nasal swab. Despite being highly effective, the test does not leave the patient risk-free and can lead to serious complications. These can be traumatic nasal cerebrospinal fluid (CSF) fistula and its consequences, such as meningitis. OBSERVATIONS: In this article, the authors present 4 case reports and a literature review. The following MeSH terms in the research were used: "CSF leak case report and covid 19." Six results were found and after searching the references and keywords 16 articles were identified. By using them, the authors tried to clarify the etiology of the fistula, its influences, and complications. LESSONS: The authors conclude that professionals must receive training, since CSF fistula originates from technical failure and lack of anatomical knowledge. The diagnosis cannot be neglected because it can bring complications to the patient's health.

Cannabis sativa L. from Seized Drug Material: In Vitro Germination and Establishment.

Background: With the expansion of the cannabis-derived product market, there is a growing need for seedling development to produce raw material for pharmaceutical applications and medicinal research. However, cannabis cultivation is illegal in many countries, and legal producers do not sell cannabis seeds in these countries. In Brazil, cannabis is still illegal, and the only way to obtain access to cannabis plants for research or as medicine is through importation, which is costly and requires authorization from the National Health Surveillance Agency (ANVISA), or from material seized by the police from drug trafficking. Methods: Therefore, since cannabis seeds obtained from drug trafficking have never been tested regarding their viability and use in in vitro cultivation, the aim of this study was to analyze the in vitro establishment of cannabis from seeds derived from Brazilian drug trafficking seizures that were provided by the police to investigate seed disinfestation procedures and further multiplication of nodal segments, with the purpose of obtaining material for medicinal research in the country. Seeds were subjected to four disinfestation treatments. Results: The best disinfestation treatment consisted in submerging the seeds in a 2 g·L-1 Captan® solution for 30 min before following the standard procedure with 70% ethanol for 30 sec and then 20 min in 2.5% sodium hypochlorite. The in vitro establishment of cannabis from seeds originating from Brazilian drug trafficking seizures was successful. The germination rate ranged from 10% to 90% according to the sample material. Non-brick weed, which consisted of dry leaves, stalks, and flowers, was more suitable for seed extraction and germination. Clones originating from BW4b showed the best development results compared with others. Conclusions: This is the first report of in vitro cannabis use in Brazil and opens great prospects for future work on its cultivation and research for medicinal applications in the country without relying on seed importation.

A new experimental model to study shrimp allergy.

Shrimp is among the most sensitizing food allergens and has been associated with many anaphylaxis reactions. However, there is still a shortage of studies that enable a systematic understanding of this disease and the investigation of new therapeutic approaches. This study aimed to develop a new experimental model of shrimp allergy that could enable the evaluation of new prophylactic treatments. BALB/c mice were subcutaneously sensitized with 100 µg of shrimp proteins of Litopenaeus vannamei adsorbed in 1 mg of aluminum hydroxide on day 0, and a booster (100 µg of shrimp proteins only) on day 14. The oral challenge protocol was based on the addition of 5 mg/ml of shrimp proteins to water from day 21 to day 35. Analysis of shrimp extract content detected at least 4 of the major allergens reported to L. vannamei. In response to the sensitization, allergic mice showed significantly enhanced IL-4 and IL-10 production in restimulated cervical draining lymph node cells. High detection of serum anti-shrimp IgE and IgG1 suggested the development of allergies to shrimp while Passive Cutaneous Anaphylaxis assay revealed an IgE-mediated response. Immunoblotting analysis revealed that Allergic mice developed antibodies to multiple antigens present in the shrimp extract. These observations were supported by the detection of anti-shrimp IgA production in intestinal lavage samples and morphometric intestinal mucosal changes. Therefore, this experimental protocol can be a tool to evaluate prophylactic and therapeutic approaches.

Negligible Effects of Nutraceuticals from Beetroot Extract on Cardiovascular and Autonomic Recovery Response following Submaximal Aerobic Exercise in Physically Active Healthy Males: A Randomized Trial.

BACKGROUND: There is little evidence that nutraceuticals from beetroot extract are beneficial with regards to recovery of the cardiovascular parameters and the autonomic nervous system (ANS) after submaximal aerobic exercise, though this formulation is employed widely for this purpose. OBJECTIVE: To study the effects of beetroot extract supplementation on the recovery of cardiorespiratory and autonomic parameters after a session of submaximal aerobic exercise. METHODS: Sixteen healthy male adults commenced a cross-over, randomized, double-blind and placebo-controlled trial. Beetroot extract (600 mg) or placebo (600 mg) were ingested 120 min prior to evaluation on randomized days. We assessed systolic blood pressure (SBP), diastolic blood pressure (DBP), pulse pressure (PP), mean arterial pressure (MAP), heart rate (HR) and HR variability (HRV) indexes at Rest and during 60 min of recovery from submaximal aerobic exercise. RESULTS: Beetroot extract ingestion slightly accelerated HR, SBP, DBP and MAP reduction following exercise associated to the placebo protocol (vs. rest). Yet no group effect (p = 0.99) was identified between the beetroot and placebo protocols on HR mean, in addition to interaction (group vs. time) (p = 0.60). No group effect was attained between the SBP (p = 0.90), DBP (p = 0.88), MAP (p = 0.73) and PP (p = 0.99) protocols and no significant differences (group vs. time) were observed between the values of SBP (p = 0.75), DBP (p = 0.79), MAP (p = 0.93) and PP (p = 0.63) between placebo and beetroot protocols. Similarly, the reoccurrence of cardiac vagal modulation after exercise via the HF (ms2) was enhanced, but not with regards to the RMSSD index. No group effect (p = 0.99) was identified for the HF (p = 0.90) and RMSSD (p = 0.67) indices. Likewise, we observed no significant differences (group vs. time) amongst the values of HF (p = 0.69) and RMSSD (p = 0.95) between the placebo and beetroot protocols. CONCLUSION: Whilst beetroot extract might assist the recovery of the cardiovascular and autonomic systems following submaximal aerobic exercise in healthy males, these results seem insignificant owing to minor differences between interventions and are weak clinically.

Improving the selection of focal species exposed to pesticides to support ecological risk assessments.

Risk assessment investigates the potential impacts of chemicals on non-target organisms. To assess the risk, ecotoxicologists study the responses of a panel of species to different substance exposure. Among the different parameters used to select indicator species (i.e. focal species), their frequency of occurrence is considered as the key parameter. Although species occurrence within a given habitat is easy to determine, we argue that it does not totally reflect the dependence of a species on a given habitat or its potential exposure to chemicals. In this study, we combined the occurrence of species with their habitat-specificity to identify focal species for risk assessment in cereals. We showed that ranking species by occurrence or by habitat-specificity produced different results, with generalist species ranking high in the occurrence list, and species with specialised habitats ranking high in the abundance list. Integrating frequency and abundance of species into one single indicator (the "Indicator Value") allows us to rank species with specialised habitats as high as generalist species in the top rank species list. Although habitat-specificity is an ecologically meaningful concept, it is largely overlooked in eco-toxicological risk assessment, despite the fact that specialists are good indicators of various environmental pressures. This method could be used extensively at different scales and could contribute to studies on risk assessment issue by (re)introducing ecological and population-level concepts and opening up new trait-based approaches.

Computerized photogrammetry used to calculate the brow position index.

BACKGROUND: The orbital region is of vital importance to facial expression. Brow ptosis, besides having an impact on facial harmony, is a sign of aging. Various surgical techniques have been developed to increase the efficacy of brow-lift surgery. However, no consensus method exists for an objective measurement of the eyebrow position due to the curvature of the face. Therefore, this study aimed to establish a method for measuring the eyebrow position using computerized photogrammetry. METHODS: For this study, 20 orbital regions of 10 volunteers were measured by direct anthropometry using a digital caliper and by indirect anthropometry (computerized photogrammetry) using standardized digital photographs. Lines, points, and distances were defined based on the position of the anthropometric landmarks endocanthion and exocanthion and then used to calculate the brow position index (BPI). Statistical analysis was performed using Student's t test with a significance level of 5 %. RESULTS: The BPI values obtained by computerized photogrammetric measurements did not differ significantly from those obtained by direct anthropometric measurements (p > 0.05). The mean BPI was 84.89 ± 10.30 for the computerized photogrammetric measurements and 85.27 ± 10.67 for the direct anthropometric measurements. CONCLUSION: The BPI defined in this study and obtained by computerized photogrammetry is a reproducible and efficient method for measuring the eyebrow position. LEVEL OF EVIDENCE III: This journal requires that authors assign a level of evidence to each article.

Determination of the Ca, Mn, Mg and Fe in honey from multiple species of stingless bee produced in Brazil.

Stingless Bees (SB) honey is a product used for different purposes, and it is worth highlighting the presence of metallic species, which can have a nutritional or toxic effect, depending on the metal present and/or the amount ingested. Therefore, the objective of this work was to quickly determine Fe, Mn, Mg and Ca in SB honey, using slurry sampling and FAAS of 88 honey samples from different regions of Brazil. The concentrations varied from < LOD to 364 µg g-1 (Ca). The order for average concentration in the samples was Ca > Mg > Mn > Fe. Through the principal component analysis, it was verified that the SB honey analyzed in the present study has higher contents of the evaluated metals than the honey of the species Apis mellifera and SB honey from other Brazilian states. Theoretical calculation demonstrated that there is a little contribution of SB honey to human diet in relation to the metals, being more significative for Mn.

The anti-cancer potential of crotoxin in estrogen receptor-positive breast cancer: Its effects and mechanism of action.

Estrogen receptor-positive (ER+) breast cancer is the most diagnosed subtype of breast cancer. Currently, aromatase inhibitors (AIs) are used as first-line treatment option in this type of tumors, however they cause several side effects, which is why new therapeutic approaches are demanding. The South American rattlesnake Crotalus durissus terrificus produces a venom enriched in several bioactive substances, like phospholipases A2 (PLA2). One of those is crotoxin, a ß-neurotoxin, that has already been reported for its anti-cancer properties in different cancers. Recently, its clinical interest has emerged and, in fact, a clinical trial in patients with advanced cancer is underway. Considering this, in this work, we studied the biological mechanisms behind the anti-cancer effects of crotoxin B (CTX) in an ER+ aromatase-overexpressing breast cancer cell line (MCF-7aro cells). Results revealed that CTX impairs MCF-7aro cells growth, through a cell cycle arrest at G2/M phase, inhibition of ERK1/2 pathway and by apoptosis through activation of caspase-8. In addition, it can be considered a safe natural compound as did not affect non-cancerous cells and only showed anti-growth effects in breast cancer cells. Therefore, this study represents an important landmark to better understand the effects and mechanisms of action of crotoxin in ER+ breast cancer.

Pattern-Reversal Visual Evoked Potential in Children With Congenital Zika Syndrome.

PURPOSE: To objectively evaluate the visual function in children with congenital Zika syndrome using pattern-reversal visual evoked potential (PR-VEP). METHODS: This was a cross-sectional study composed of two patient groups: children aged between 18 and 24 months who were positive for the Zika virus (congenital Zika syndrome group) and age-matched healthy children (control group). All patients underwent a comprehensive ophthalmologic examination and PR-VEP was performed binocularly in a room with constant dim illumination. The P100 peak time in the 60' arc stimuli was used, and the results were correlated with visual acuity, cephalic perimeter at birth, and funduscopic findings. RESULTS: Thirty-seven children were included in the congenital Zika syndrome group and 15 in the control group. The mean age was 18.5 ± 0.9 months (range: 17 to 20 months) in the congenital Zika syndrome group and 24.3 ± 1.6 months (range: 21 to 28 months) in the control group. The P100 response was normal in 7 patients (18.9%) with congenital Zika syndrome, borderline in 2 (5.4%), abnormal with prolonged latency in 18 (48.6%), and abnormal with no response in 10 (27.0%). A significant correlation between the visual acuity and P100 peak time was observed (P < .001). The P100 values were not correlated significantly with the cephalic perimeter at birth (P = .412) or the funduscopic findings (P = .510). PR-VEP in children with congenital Zika syndrome and no funduscopic findings was significantly worse than in the control group (P = .001). CONCLUSIONS: Children with congenital Zika syndrome have characteristically abnormal PR-VEPs regardless of the funduscopic findings and severe microcephaly. The PR-VEP findings supported the diagnosis of cortical visual impairment in these children. [J Pediatr Ophthalmol Strabismus. 2021;58(2):78-83.].

A real-world implementation of a nationwide, long-term monitoring program to assess the impact of agrochemicals and agricultural practices on biodiversity.

Biodiversity has undergone a major decline throughout recent decades, particularly in farmland. Agricultural practices are recognized to be an important pressure on farmland biodiversity, and pesticides are suspected to be one of the main causes of this decline in biodiversity. As part of the national plan for reduction of pesticides use (Ecophyto), the French ministry of agriculture launched the 500 ENI (nonintended effects) monitoring program in 2012 in order to assess the unintended effects of agricultural practices, including pesticide use, on biodiversity represented by several taxonomic groups of interest for farmers. This long-term program monitors the biodiversity of nontargeted species (earthworms, plants, coleoptera, and birds), together with a wide range of annual data on agricultural practices (crop rotation, soil tillage, weed control, fertilizers, chemical treatments, etc.). Other parameters (e.g., landscape and climatic characteristics) are also integrated as covariates during the analyses. This monitoring program is expected to improve our understanding of the relative contribution of the different drivers of population and community trends. Here, we present the experience of setting up the 500 ENI network for this ambitious and highly complex monitoring program, as well as the type of data it collects. The issue of data quality control and some first results are discussed. With the aim of being useful to readers who would like to set up similar monitoring schemes, we also address some questions that have arisen following the first five years of the implementation phase of the program.

Efficient photodynamic inactivation of Leishmania parasites mediated by lipophilic water-soluble Zn(II) porphyrin ZnTnHex-2-PyP4.

BACKGROUND: Photodynamic inactivation (PDI) is emerging as a promising alternative for cutaneous leishmaniasis (CL). The chemotherapy currently used presents adverse effects and cases of drug resistance have been reported. ZnTnHex-2-PyP4+ is a porphyrin with a high potential as a photosensitizer (PS) for PDI, due to its photophysical properties, structural stability, and cationic/amphiphilic character that can enhance interaction with cells. This study aimed to investigate the photodynamic effects mediated by ZnTnHex-2-PyP4+ on Leishmania parasites. METHODS: ZnTnHex-2-PyP4+ stability was evaluated using accelerated solvolysis conditions. The photodynamic action on promastigotes was assessed by (i) viability assays, (ii) mitochondrial membrane potential evaluation, and (iii) morphological analysis. The PS-promastigote interaction was studied. PDI on amastigotes and the cytotoxicity on macrophages were also analyzed. RESULTS: ZnTnHex-2-PyP4+, under submicromolar concentration, led to immediate inactivation of more than 95% of promastigotes. PDI promoted intense mitochondrial depolarization, loss of the fusiform shape, and plasma membrane wrinkling in promastigotes. Fluorescence microscopy revealed a punctate PS labeling in the parasite cytoplasm. PDI also led to reductions of ca. 64% in the number of amastigotes/macrophage and 70% in the infection index after a single treatment session. No noteworthy toxicity was observed on mammalian cells. CONCLUSIONS: ZnTnHex-2-PyP4+ is stable against demetallation and more efficient as PS than the ethyl analogue ZnTE-2-PyP4+, indicating readiness for evaluation in in vivo studies as an alternative approach to CL. GENERAL SIGNIFICANCE: This report highlighted promising photodynamic effects mediated by ZnTnHex-2-PyP4+ on Leishmania parasites, opening up perspectives for applications in CL pre-clinical assays and PDI of other microorganisms.

Oral Tolerance Induced by Heat Shock Protein 65-Producing Lactococcus lactis Mitigates Inflammation in Leishmania braziliensis Infection.

Cutaneous leishmaniasis caused by L. braziliensis induces a pronounced Th1 inflammatory response characterized by IFN-γ production. Even in the absence of parasites, lesions result from a severe inflammatory response in which inflammatory cytokines play an important role. Different approaches have been used to evaluate the therapeutic potential of orally administrated heat shock proteins (Hsp). These proteins are evolutionarily preserved from bacteria to humans, highly expressed under inflammatory conditions and described as immunodominant antigens. Tolerance induced by the oral administration of Hsp65 is capable of suppressing inflammation and inducing differentiation in regulatory cells, and has been successfully demonstrated in several experimental models of autoimmune and inflammatory diseases. We initially administered recombinant Lactococcus lactis (L. lactis) prior to infection as a proof of concept, in order to verify its immunomodulatory potential in the inflammatory response arising from L. braziliensis. Using this experimental approach, we demonstrated that the oral administration of a recombinant L. lactis strain, which produces and secretes Hsp65 from Mycobacterium leprae directly into the gut, mitigated the effects of inflammation caused by L. braziliensis infection in association or not with PAM 3CSK4 (N-α-Palmitoyl-S-[2,3-bis(palmitoyloxy)-(2RS)-propyl]-L-cysteine, a TLR2 agonist). This was evidenced by the production of anti-inflammatory cytokines and the expansion of regulatory T cells in the draining lymph nodes of BALB/c mice. Our in vitro experimental results suggest that IL-10, TLR-2 and LAP are important immunomodulators in L. braziliensis infection. In addition, recombinant L. lactis administered 4 weeks after infection was observed to decrease lesion size, as well as the number of parasites, and produced a higher IL-10 production and decrease IFN-γ secretion. Together, these results indicate that Hsp65-producing L. lactis can be considered as an alternative candidate for treatment in both autoimmune diseases, as well as in chronic infections that cause inflammatory disease.

Evaluation of microbial contamination in multi-dose fluorescein eyedrops in a reference eye center.

PURPOSE: To analyze the presence of microorganisms in fluorescein eyedrops used in a reference eye center in Recife-PE. METHODS: This real-life and masked study evaluated fluorescein eyedrops used at the Altino Ventura Foundation in May 2019. Cultures were performed according to exposure times; I) three eyedrop bottles were analyzed after one day of use, II) three eyedrop bottles after 4 d of use, III) three eyedrop bottles after 8 d of use, and IV) three unopened bottles used as control. Samples were collected from the bottle's tip, instilled drop, and residual fluid. After incubation, all colonies were analyzed and identified through biochemical tests. RESULTS: The contamination rate of the fluorescein eyedrop bottles in this study was 55.5% (5/9 vials). There was no contamination in the control group. The highest contamination was seen in one day exposed eyedrops, in 100% of the bottles. The bottle's tip had a higher rate of contamination compared to the drop and residual fluid. Gram-positive bacteria were isolated in 7/27 (25.9%) samples. Growth of fungi or gram-negative bacteria was not observed. CONCLUSION: The identification of gram-positive bacteria predominantly on the tip of the fluorescein eyedrop bottles suggests inadequate handling as the main cause of contamination.