Wastewater surveillance for viral pathogens: A tool for public health.

A focus on water quality has intensified globally, considering its critical role in sustaining life and ecosystems. Wastewater, reflecting societal development, profoundly impacts public health. Wastewater-based epidemiology (WBE) has emerged as a surveillance tool for detecting outbreaks early, monitoring infectious disease trends, and providing real-time insights, particularly in vulnerable communities. WBE aids in tracking pathogens, including viruses, in sewage, offering a comprehensive understanding of community health and lifestyle habits. With the rise in global COVID-19 cases, WBE has gained prominence, aiding in monitoring SARS-CoV-2 levels worldwide. Despite advancements in water treatment, poorly treated wastewater discharge remains a threat, amplifying the spread of water-, sanitation-, and hygiene (WaSH)-related diseases. WBE, serving as complementary surveillance, is pivotal for monitoring community-level viral infections. However, there is untapped potential for WBE to expand its role in public health surveillance. This review emphasizes the importance of WBE in understanding the link between viral surveillance in wastewater and public health, highlighting the need for its further integration into public health management.

IL-27-engineered CAR.19-NK-92 cells exhibit enhanced therapeutic efficacy.

Chimeric antigen receptor (CAR) engineering of natural killer (NK) cells has shown promising results in early-phase clinical studies. However, advancing CAR-NK cell therapeutic efficacy is imperative. In this study, we investigated the impact of a fourth-generation CD19-targeted CAR (CAR.19) coexpressing IL-27 on NK-92 cells. We observed a significant improvement in NK-92 cell proliferation and cytotoxicity activity against B-cell cancer cell lines, both in vitro and in a xenograft mouse B-cell lymphoma model. Our systematic transcriptome analysis of the activated NK-92 CAR variants further supports the potential of IL-27 in fourth-generation CARs to overcome limitations of NK cell-based targeted tumor therapies by providing essential growth and activation signals. Integrating IL-27 into CAR-NK cells emerges as a promising strategy to enhance their therapeutic potential and elicit robust responses against cancer cells. These findings contribute substantially to the mounting evidence supporting the potential of fourth-generation CAR engineering in advancing NK cell-based immunotherapies.

Influence of crown-lengthening surgery on teeth rehabilitated with ceramic single crowns: A 12-month prospective clinical study.

OBJECTIVE: To compare the clinical periodontal parameters of teeth restored with a single ceramic crown, with and without crown lengthening procedure. METHODS: This prospective, longitudinal, controlled, and single-blinded clinical trial involved 22 patients with a total of forty-one teeth with ceramic crowns. The teeth were divided into two groups: test (n = 21), comprising teeth rehabilitated post crown-lengthening surgery, and control (n = 20), comprising teeth rehabilitated without crown-lengthening surgery. Plaque index (PI), gingival index (GI), probing depth (PD), bleeding on probing (BoP), and clinical attachment level (CAL) were compared between groups (surgically treated and non-surgically treated) and within each group for each type of site (treated -tt; adjacent - ad; and nonadjacent - nad). Additionally, gingival phenotype (GP), gingival recession (GR), and keratinized tissue width (KTW) were also assessed post- restoration. Statistical analyses used a significance level set at 5 %. RESULTS: PI, GI, and BoP were reduced, but no statistically significant differences were observed within each group or between groups for most follow-up periods. CAL of the TT sites was consistently higher in the test group, and PD was also higher in the test group (p < 0.05), except at T3. adPD, nadPD, adCAL, and nadCAL demonstrated no significant differences between groups and periods. A significant association was identified between GP and the occurrence of GR, with the thick-flat phenotype demonstrating less association with GR, regardless of whether crown lengthening was performed or not. CONCLUSION: Crown-lengthening surgery in rehabilitated teeth does not significantly affect PI and GI after 12 months. Although crown-lengthening surgery affected PD and CAL in TT sites, it did not affect adjacent and non-adjacent sites. CLINICAL RELEVANCE: These findings emphasize the importance of considering individual patient factors and the potential impact on periodontal tissues when planning crown-lengthening surgery. Clinicians must have a comprehensive understanding of the dynamics of the periodontal tissues involved in restorative treatments to optimize the procedure, increase success rates, and minimize potential complications.

Gametogenesis and seminatural reproduction of the Amazon twospot astyanax Astyanax bimaculatus (Linnaeus, 1758) cultivated in an enriched environment.

Environmental enrichment is used to provide well-being to the animals, such as fish, in captive conditions, mimicking their natural habitat. It may influence fish behavior, physiology, and survival. In terms of reproduction, however, the relationship between environment enrichment and successful reproduction in captivity is still poorly explored in fish species. Aiming to understand any possible benefits of structural enrichment on fish reproduction, 10-days-hatched larvae of the twospot astyanax Astyanax bimaculatus were raised for 18 weeks in tanks with different elements of structural environmental enrichment (PVC pipes, stones, and artificial plants). In the 5th month of life, those animals were hormonally induced to reproduce to assess gamete formation and offspring quality. Animals raised in a sterile-reared environment (non-enriched) showed earlier spawning than the enriched one, presenting significant quantities of Postovulatory follicle complexes (POCs) and cells in atresia in female ovaries, indicating possible reproductive dysfunction or stress, as well as a greater quantity of empty testicular lumen in males, indicating great release of sperm. On the contrary, animals cultivated in enriched environments showed gonads filled with semen in males and vitellogenic oocytes in females. Furthermore, offspring from the sterile-reared group presented significant rates of larval abnormality compared to the enriched group. In conclusion, the results of this study show that environmental enrichment can interfere with the reproduction of fish in captivity, mainly by preventing early maturation of gametes, which can result in low-quality offspring and, consequently, low production of fish species.

Negative impacts of social isolation on behavior and neuronal functions are recovered after short-term social reintroduction in zebrafish.

Recently, social isolation measures were crucial to prevent the spread of the coronavirus pandemic. However, the lack of social interactions affected the population mental health and may have long-term consequences on behavior and brain functions. Here, we evaluated the behavioral, physiological, and molecular effects of a social isolation (SI) in adult zebrafish, and whether the animals recover such changes after their reintroduction to the social environment. Fish were submitted to 12 days of SI, and then reintroduced to social context (SR). Behavioral analyses to evaluate locomotion, anxiety-like and social-related behaviors were performed after SI protocol, and 3 and 6 days after SR. Cortisol and transcript levels from genes involved in neuronal homeostasis (c-fos, egr, bdnf), and serotonergic (5-HT) and dopaminergic (DA) neurotransmission (thp, th) were also measured. SI altered social behaviors in zebrafish such as aggression, social preference, and shoaling. Fish submitted to SI also presented changes in the transcript levels of genes related to neural activity, and 5-HT/DA signaling. Interestingly, most of the behavioral and molecular changes induced by SI were not found again 6 days after SR. Thus, we highlight that SR of zebrafish to their conspecifics played a positive role in social behaviors and in the expression of genes involved in different neuronal signaling pathways that were altered after 12 days of SI. This study brings unprecedented data on the effects of SR in the recovery from SI neurobehavioral alterations, and reinforces the role of zebrafish as a translational model for understanding the neurobiological mechanisms adjacent to SI and resocialization.

Leptospira borgptersenii and Leptospira interrogans identified in wild mammals in Rio Grande do Sul, Brazil.

Leptospira spp. are bacteria responsible for leptospirosis, a zoonotic disease with considerable impacts on the economy, animal health, and public health. This disease has a global distribution and is particularly prevalent in Brazil. Both rural and urban environments are habitats for Leptospira spp., which are primarily transmitted through contact with the urine of infected animals. Consequently, domestic and wild species can harbor these prokaryotes and serve as infection sources for other hosts. In the context of wild animals, there is a dearth of molecular studies elucidating the roles of various animal and bacterial species in the epidemiology of leptospirosis. Therefore, this study aimed to evaluate the presence of Leptospira spp. DNA in different species of free-living and captive wild animals and to assess the phylogenetic relationships of the identified microorganisms in Rio Grande do Sul, Brazil. The samples were evaluated for the presence of the gene lipL32 by polymerase chain reaction (PCR) and sequencing of the amplified fragment after which phylogenetic analyzes were carried out. DNA from Leptospira spp. was extracted from kidney tissue from wild animals (Mammalia class). Pathogenic Leptospira spp. DNA was detected in 9.6% (11/114) of the samples, originating from nine species of wild animals, including the white-eared opossum (Didelphis albiventris), skunk (Conepatus chinga), geoffroy's cat (Leopardus geoffroyi), margay (Leopardus wiedii), pampas fox (Lycalopex gymnocercus), capybara (Hydrochoerus hydrochaeris), common marmoset (Callithrix jacchus), neotropical river otter (Lontra longicaudis), and european hare (Lepus europaeus). Phylogenetic analysis revealed the presence of Leptospira borgpetersenii and Leptospira interrogans in these animals. This research is the first study contributing to the epidemiology of leptospirosis by identifying L. borgpetersenii and L. interrogans in free-living and captive wild animals in Rio Grande do Sul, Brazil, potentially acting as bacterial reservoirs. Additionally, our findings can inform sanitary measures for controlling and preventing the disease, thereby safeguarding public health.

Precarious work and methodological challenges to study hard-to-reach populations.

OBJECTIVE: To describe the methodological challenges and strategies of a web survey on the working conditions and health among delivery workers. METHODS: The study population consisted of Brazilian delivery workers operating in the national territory. Procedures include building solid and ongoing collaboration with worker representatives and conducting a four-month data collection from February to May 2022, sharing the link to the online questionnaire on social media such as social networks (Facebook, Instagram) and messaging apps (WhatsApp, Telegram). RESULTS: The recruitment of 41 leaders or influencers of delivery workers increased the dissemination of the study, some of whom participated in the consensual validation of the questionnaire; the production of content for social media for the dissemination of the questionnaire link on social networks and applications, and the in-person dissemination of the study at the delivery workers' meeting points during the workday played a fundamental role, totaling around 132 hours in 45 shifts. The strategies adopted for data collection with a hybrid approach to dissemination made it possible to carry out the web survey. After four months of the web survey, 564 delivery workers, 543 men and 18 women, responded to the online questionnaire. CONCLUSION: The web survey presented methodological strategies to overcome the challenge of reaching workers, including hybrid work, to increase the participation of workers, on whom epidemiological research is still scarce.

Detection of pathogenic Leptospira spp. in unconventional pets.

Leptospirosis is a disease caused by Leptospira spp. responsible for considerable impacts on the public and animal health. In the past two decades, non-domesticated species of pets (unconventional pets) have become popular. However, the role of these unconventional pets on maintaining diseases still unclear. Therefore, the objective of this study was to survey the presence of Leptospira spp. DNA in unconventional pets. Samples of kidney tissues from 29 animals belonging to the Mammalia class (including Orders Carnivora, Lagomorpha and Rodentia) were analyzed for the presence of the gene lipL32. As a result, DNA of pathogenic Leptospira spp. from specie L. interrogans was detected in four (13,80%) of the analyzed samples: three from Oryctolagus cuniculus and one from Mesocricetus auratus. This study highlights the importance of epidemiological surveillance of leptospirosis, as it identified in species of unconventional pets, that may possibly act as reservoirs of Leptospira spp.

Antioxidant responses driven by Hesperetin and Hesperidin counteract Parkinson's disease-like phenotypes in Drosophila melanogaster.

The study investigated the protective effects of Hesperetin (HSP) and Hesperidin (HSD) on 1 methyl, 4 phenyl, 1,2,3,6 tetrahydropyridine hydrochloride (MPTP)-induced Parkinsonism in Drosophila melanogaster (D. melanogaster). After a lifespan study to select exposure time and concentrations, flies were co-exposed to MPTP (0.4 mg/g diet), Hesperetin (0.2 and 0.4 mg/g diet), and Hesperidin (0.1 and 0.4 mg/g) for 7 days. In addition to in vivo parameters, we assayed some markers of oxidative stress and antioxidant status (lipid peroxidation, protein carbonylation, thiol content, hydrogen peroxide, and nitrate/nitrite levels, mRNA expression of Keap-1 (Kelch-like ECH associated protein 1), /Nrf2 (Nuclear factor erythroid 2 related factor 2), catalase, and glutathione-S-transferase (GST) activities), and cholinergic (acetyl cholinesterase activity (AChE) and dopaminergic signaling content and the mRNA expression of tyrosine hydroxylase (TH), monoamine oxidase (MAO-like) activity). In addition to increasing the lifespan of flies, we found that both flavonoids counteracted the adverse effects of MPTP on survival, offspring emergence, and climbing ability of flies. Both flavonoids also reduced the oxidative damage on lipids and proteins and reestablished the basal levels of pro-oxidant species and activities of antioxidant enzymes in MPTP-exposed flies. These responses were accompanied by the normalization of the mRNA expression of Keap1/Nrf2 disrupted in flies exposed to MPTP. MPTP exposure also elicited changes in mRNA expression and content of TH as well as in MAO and AChE activity, which were reversed by HST and HSD. By efficiently hindering the oxidative stress in MPTP-exposed flies, our findings support the promising role of Hesperetin and Hesperidin as adjuvant therapy to manage Parkinsonism induced by chemicals such as MPTP.

JM-20 potently prevents the onset of caffeine-induced anxiogenic phenotypes in zebrafish (Danio rerio).

Anxiety is among the most prevalent mental disorders present in the general population. Benzodiazepines are the most commonly prescribed drugs for the treatment of anxiety. Using zebrafish as a model organism, we investigated the anxiolytic activity of JM-20, a novel hybrid molecule with a 1,5-benzodiazepine ring fused to a dihydropyridine moiety. Firstly, we carried out some assays to analyze the possible toxicity mediated by JM-20. For this, zebrafish were exposed to different JM-20 concentrations (0-5 µM) for 96 h. Then, using the novel tank test, we evaluated both locomotor and anxiety-like behavior of the animals. Furthermore, brain, liver and plasma were removed to assess toxicity parameters. JM-20 exposure did not cause changes on novel tank, and also did not alter brain viability, hepatic LDH and plasma ALT levels. Afterward, we investigated whether a pre-exposure to JM-20 would prevent the anxiogenic effect evoked by caffeine. In the novel tank test, caffeine significantly decreased the time spent at the top, as well as the number of transitions to the top area. Moreover, caffeine decreased both the total and average time spent in the lit area, as well as increased the number of risk episodes evaluated by the light-dark test. Whole-body cortisol levels were also increased by caffeine exposure. Interestingly, pre-treatment with JM-20 abolished all alterations induced by caffeine. The anxiolytic effect profile of JM-20 was similar to those found for diazepam (positive control). Our findings show, for the first time, the anxiolytic effect of JM-20 in zebrafish, and its relationship with cortisol regulation.

Precarious work and methodological challenges to study hard-to-reach populations

ABSTRACT OBJECTIVE To describe the methodological challenges and strategies of a web survey on the working conditions and health among delivery workers. METHODS The study population consisted of Brazilian delivery workers operating in the national territory. Procedures include building solid and ongoing collaboration with worker representatives and conducting a four-month data collection from February to May 2022, sharing the link to the online questionnaire on social media such as social networks (Facebook, Instagram) and messaging apps (WhatsApp, Telegram). RESULTS The recruitment of 41 leaders or influencers of delivery workers increased the dissemination of the study, some of whom participated in the consensual validation of the questionnaire; the production of content for social media for the dissemination of the questionnaire link on social networks and applications, and the in-person dissemination of the study at the delivery workers' meeting points during the workday played a fundamental role, totaling around 132 hours in 45 shifts. The strategies adopted for data collection with a hybrid approach to dissemination made it possible to carry out the web survey. After four months of the web survey, 564 delivery workers, 543 men and 18 women, responded to the online questionnaire. CONCLUSION The web survey presented methodological strategies to overcome the challenge of reaching workers, including hybrid work, to increase the participation of workers, on whom epidemiological research is still scarce.

GXMR-CAR containing distinct GXM-specific single-chain variable fragment (scFv) mediated the cell activation against Cryptococcus spp. And had difference in the strength of tonic signaling.

Cryptococcus spp. has a polysaccharide capsule composed of glucuronoxylomannan-GXM, a major virulence factor that can prevent the recognition of fungi by immune cells. Chimeric Antigen Receptor (CAR) redirects T cells to target Cryptococcus spp. as previously demonstrated by a CAR specific to GXM, GXMR-CAR. The current study evaluated the strength of the signal transduction triggered by GXMR-CAR, composed of a distinct antigen-binding domain sourced from a single-chain variable fragment (scFv). GXM-specific scFv derived from mAbs 2H1 and 18B7, 2H1-GXMR-CAR and 18B7-GXMR-CAR, respectively, were designed to express CD8 molecule as hinge/transmembrane, and the costimulatory molecule CD137 (4-1BB) coupled to CD3ζ. The 2H1-GXMR-CAR or 18B7-GXMR-CAR Jurkat cells recognized soluble GXM from C. gattii and C. neoformans, and the levels of IL-2 released by the modified cells did not differ between the GXMR-CAR constructs after exposure to Cryptococcus spp. 18B7-GXMR-CAR triggered tonic signaling was more pronounced in modified Jurkat cells, and a protein kinase inhibitor of the Src family (dasatinib) significantly reduced GXMR-CAR tonic signaling and inhibited cell activation against ligands. 18B7 scFv showed a structural modification of the variable heavy (VH) chain that clarified the difference in the strength of tonic signaling and the level of cell activation between 2H1-GXMR-CAR and 18B7-GXMR-CAR. GXMR-CAR constructs induced T-cell activation against clinical isolates of Cryptococcus spp. and serum from patients with cryptococcosis induced high levels of IL-2, mainly in cells modified with 18B7-GXMR-CAR. Thus, 18B7-GXMR-CAR and 2H1-GXMR-CAR mediated T cell activation against Cryptococcus spp. and 18B7 and 2H1 scFv influenced the strength of tonic signaling.

2H1-GXMR-CAR and 18B7-GXMR-CAR are efficiently expressed on the cell surface;2H1-GXMR-CAR and 18B7-GXMR-CAR redirected T cells toward the ligands;18B7-GXMR-CAR provided highest levels of tonic signaling;Binding pocket of 18B7 scFv favored the tonic signaling triggered by GXMR-CAR;Binding pocket of 18B7 scFv favored the tonic signaling triggered by GXMR-CAR.

Long-term monitoring of COVID-19 prevalence in raw and treated wastewater in Salvador, the largest capital of the Brazilian Northeast.

Wastewater-based epidemiology (WBE) becomes an interesting epidemiological approach to monitoring the prevalence of SARS-CoV-2 broadly and non-invasively. Herein, we employ for the first time WBE, associated or not with the PEG 8000 precipitation method, for the detection of SARS-CoV-2 in samples of raw or treated wastewater from 22 municipal wastewater treatment stations (WWTPs) located in Salvador, the fourth most populous city in Brazil. Our results demonstrate the success of the application of WBE for detecting SARS-CoV-2 in both types of evaluated samples, regardless of the usage of PEG 8000 concentration procedure. Further, an increase in SARS-CoV-2 positivity rate was observed in samples collected in months that presented the highest number of confirmed COVID-19 cases (May/2021, June/2021 and January/2022). While PEG 8000 concentration step was found to significantly increase the positivity rate in treated wastewater samples (p < 0.005), a strong positive correlation (r: 0.84; p < 0.002) between non-concentrated raw wastewater samples with the number of new cases of COVID-19 (April/2021-February/2022) was observed. In general, the present results reinforce the efficiency of WBE approach to monitoring the presence of SARS-CoV-2 in either low- or high-capacity WWTPs. The successful usage of WBE even in raw wastewater samples makes it an interesting low-cost tool for epidemiological surveillance.

Anti-Leishmania spp. antibody detection in domestic cats from a visceral leishmaniasis transmission area.

Feline leishmanial infection is reported worldwide, but the epidemiological role of domestic cats in the leishmaniasis cycle remains unclear, and cats might act as cryptic reservoir hosts in endemic areas with no feline leishmaniosis cases. Considering that, a serological screening for anti-Leishmania spp. antibodies was performed by indirect immunofluorescence antibody test (IFAT) in 389 necropsied cats' serum samples from a new visceral leishmaniasis transmission area with no feline leishmanial infection reported to unveil if the cats are being exposed to the parasite. The overall seroprevalence for Leishmania spp. was 11.05% (43/389). No association was found between sex, neutering status, age group, breed, coat length, feline immunodeficiency virus (FIV) infection, and Leishmania spp. antibody detection. A positive association was found with coat color (cats within the orange spectrum with white [particolor]) (OR = 2.47, CI 95% 1 - 6.13, P = 0.044) and a negative association (OR = 0.38, CI 95% 0.18 - 0.79, P = 0.01) between feline leukemia virus (FeLV) infection and IFAT positivity for Leishmania spp. Therefore, it is concluded that the seroprevalence found was greater than 10%, indicating contact of the protozoan with cats in the region served.

EtHg is more toxic than MeHg to human peripheral blood mononuclear cells: Involvement of apoptotic, mitochondrial, oxidative and proliferative parameters.

BACKGROUND: Methylmercury (MeHg) and ethylmercury (EtHg) are potent toxicants affecting the environment and human healthy. In this way, the present study aimed to investigate and compare the effects of MeHg and EtHg exposure on human peripheral blood mononuclear cells (PBMCs), which are critical components of the mammalian immune system. METHODS: PBMCs were exposed to 2.5 µM MeHg or 2.5 µM EtHg. The number of cells and incubation times varied according to each assay. After exposures, the PBMCs were subjected to different evaluations, including cell viability, morphological aspects, cell cycle phases, indices of apoptosis and necrosis, reactive species (RS) production, and mitochondrial functionality. RESULTS: PBMCs exposed to EtHg were characterized by decreased viability and size, increased granularity, RS production, and apoptotic indexes accompanied by an intensification of Sub-G1 and reduction in G0-G1 cell cycle phases. Preceding these effects, we found mitochondrial dysfunctions, namely a reduction in the electron transport system related to mitochondrial complex I. In contrast, PBMCs exposed to MeHg showed only reduced viability. By ICP-MS, we found that PBMCs treated with EtHg accumulated Hg + levels ∼1.8-fold greater than MeHg-exposed cells. CONCLUSIONS AND SIGNIFICANCE: Taken together, our findings provide important insights about mercury immunotoxicity, showing that EtHg is more immunotoxic to human PBMCs than MeHg.

Injuries and complaints in the Brazilian national volleyball male team: a case study.

BACKGROUND: The literature reports a vast amount of epidemiological information on injuries in volleyball athletes. However, little is known about the incidence of injuries in elite athletes of international level participating in major competitions, such as world championships and Olympic games. The objective of the study was to analyse the incidence of injuries in elite professional volleyball athletes, and the prevalence of complaints reported by athletes. METHODS: This is a case study in which data were collected between April 2018 and August 2021. All the athletes called to play for the Brazilian national male volleyball team during the analysis period participated. From the athletes' medical records, the occurrence of injuries (injurious events that lead to a time off from activities) and complaints (discomforts that did not lead to a time off from activities) were analysed. Frequency data were used to calculate incidence, prevalence and ratios. RESULTS: From 41 athletes who played for the team during the analysed period, 12 athletes had 28 injuries and 38 athletes reported 402 complaints. For injuries, an incidence of seven injuries/1,000 h of competition and two injuries/1,000 h of training was observed. The average recovery time of the athletes was 10 days. The regions with the highest prevalence of injuries were the knee (111/1,000 athletes) and ankle (69/1,000 athletes). For complaints, 402 complaints required 1,085 treatment sessions, with the regions with the highest prevalence of complaints being the knee (261/1,000 complaints) followed by the shoulders (236/1,000 complaints). Athletes aged above 23 years and those playing as middle blockers and outside hitters presented a higher prevalence of injuries and complaints. CONCLUSIONS: Almost one-third of the athletes had injuries and almost all athletes reported complaints during the study period. Injuries and complaints were more prevalent in the knees. Complaints caused a high demand for the healthcare team. To manage risk of injuries for overload, specific injury prevention strategies are needed and should be included as an essential component of the training plan for elite volleyball players.

Type 2 diabetes mellitus in obesity promotes prolongation of cardiomyocyte contractile function, impaired Ca2+ handling and protein carbonylation damage.

AIMS: To investigate whether the obesity associated to T2DM presented cardiomyocyte myocardial contractility dysfunction due to damage in Ca2+ handling, concomitantly with increased biomarkers of oxidative stress. METHODS: Male Wistar rats were randomized into two groups: control (C): fed with standard diet; and obese (Ob) that fed a saturated high-fat. After the characterization of obesity (12 weeks), the Ob animals were submitted to T2DM induction with a single dose of intraperitoneal (i.p.) injection of streptozotocin (30 mg/kg). Thus, remained Ob rats that were characterized as to the presence (T2DMOb; n = 8) and/or absence (Ob; n = 10) of T2DM. Cardiac remodeling was measured by post-mortem morphological, isolated cardiomyocyte contractile function, as well as by intracellular Ca2+-handling analysis. RESULTS: T2DMOb presented a significant reduction of all fat pads, total body fat and adiposity index. T2DMOb group presented a significant increase in protein carbonylation and superoxide dismutase (SOD) activity, respectively. T2DMOb promoted elevations in fractional shortening (15.6 %) and time to 50 % shortening (5.8 %), respectively. Time to 50 % Ca2+ decay was prolonged in T2DMOb, suggesting a possible impairment in Ca2+recapture and/or removal. CONCLUSION: Type 2 diabetes mellitus in obesity promotes prolongation of cardiomyocyte contractile function with protein carbonylation damage and impaired Ca2+ handling.

On the effect of social cue valence in contextual memory persistence.

Social cues are valuable sensorial stimuli to the acquisition and retrieval of contextual memories. Here, we asked whether the valence of social cues would impact the formation of contextual memories. Adult male C57/BL6 mice were exposed to either conditioned place preference (CPP) or avoidance (CPA). As positive stimuli we used social interaction with a female (IF), while interaction with a male CD1 mice (IM) was used as negative stimulus. Contextual memory was tested 24 h and 7 days after conditioning. Aggressive behavior of CD1, as well as interaction with the female were quantified along the conditioning sessions. IM, but not IF, was salient enough to induce contextual memory estimated by the difference between the time in the conditioned context during test and habituation. Next, we chose two odors with innate behavioral responses and opposite valence to narrow down the sociability to one of its sensorial sources of information - the olfaction. We used urine from females in proestrus (U) and 2,4,5-trimethyl thiazoline (TMT), a predator odor. TMT decreased and U increased the time in the conditioned context during the test performed 24 h and 7 days after conditioning. Taken together, our results suggest that contextual memories conditioned to social encounters are difficult to stablish in mice, specially the one with positive valence. On the other hand, using odors with ecological relevance is a promising strategy to study long-term contextual memories with opposite valences. Ultimately, the behavioral protocol proposed here offers the advantage of studying contextual memories with opposite valences using unconditioned stimulus from the same sensorial category such as olfaction.

Effect of Different Frequencies of Electroacupuncture on Chronic Low Back Pain in Older Adults: A Triple-blind, Placebo-controlled, Randomized Clinical Trial.

BACKGROUND: Acupuncture is one of the most widely used therapies to treat chronic low back pain, whose analgesic effect seems to be potentiated by the addition of electric current (electroacupuncture). However, we are not aware of any clinical trial that has evaluated the effectiveness of this technique in adults > 65 years. OBJECTIVE: To evaluate the effect of electroacupuncture in the treatment of chronic low back pain in the elderly. STUDY DESIGN: Triple-blind, controlled, and randomized clinical trial. SETTING: Faculty of Medicine, University of São Paulo (USP); Sector of Biological Sciences - Physiotherapy Course, Federal University of Paraná (UFPR). METHODS: The study included 125 elderly people with chronic nonspecific low back pain who were randomized to one of 5 study groups: 3 of electroacupuncture; one control; and one placebo, all of them treated for 5 weeks. The primary endpoint was pain intensity, and secondary endpoints included the qualitative aspect of pain, functional disability (Roland-Morris and sit and stand test), emotional functioning (depression and anxiety), and psychosocial factors. Data analysis followed the intention-to-treat principle. The confidence interval was set at 95% and the significance level at 5%. RESULTS: All groups achieved a reduction in pain intensity; however, a significant difference was only detected between electroacupuncture and placebo, where the latter showed greater pain reduction. Regarding secondary outcomes, all groups showed good posttreatment results for all assessments but without statistical significance. Among the groups, the placebo was the one that obtained the best results between the pre- and post-treatment for depression, qualitative aspect of pain, and functional disability, but only for the qualitative aspect of pain and for the sit-and-stand test was a significant reduction found in the intergroup comparison. The analysis of the overall effect perceived by the participants in relation to low back pain revealed that individuals from all groups felt close to full recovery. LIMITATIONS: Absence of follow-up and a relatively small number of patients. CONCLUSIONS: This study provides evidence that there is no one frequency of electroacupuncture that is most effective in treating chronic low back pain in the elderly and that electroacupuncture is not superior to manual acupuncture or placebo treatment.

Kaempferol counteracts toxicity induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine in D. melanogaster: An implication of its mitoprotective activity.

The current study aimed to investigate whether kaempferol (KMP), the major bioactive component of green leafy vegetables, could counteract the toxicity elicited by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in Drosophila melanogaster or not. First, we performed a dose-response curve, where adult wild-type flies were fed on diet-containing different concentrations of KMP throughout their lifespan. Afterward, flies were fed on a diet containing MPTP (500 µM) and KMP (20 and 40 µM) for 7 days. The MPTP- fed flies presented a higher mortality rate, lower emergence rate, locomotor deficits, and disruption in circadian rhythm when compared to the control. MPTP exposure induced severe oxidative stress, which was marked by reduction in thiol content, overproduction of reactive species, lipid and protein oxidation, and disruption of enzymes of antioxidant and neurotransmission pathways. MPTP also compromised the mitochondrial dynamics and respiration of flies, affecting the electron transport chain, oxidative phosphorylation, and fusion/fission processes. Besides extending per se the lifespan of flies, KMP counteracted the toxic effects of MPTP on the circadian cycle, survival, climbing, and hatching rates. KMP was also effective in restoring the activities of acetylcholinesterase (AChE) and monoamine oxidase (MAO) enzymes, as well as in normalizing the levels of all oxidant/antioxidant markers disrupted in MPTP-fed flies. Indeed, KMP reestablished the mitochondrial functionality in MPTP- fed flies, restoring the electron transport system linked to mitochondrial complex I and II, and rescuing the mRNA transcription of genes associated with mitochondrial fusion and fission, namely OPA-1 (Optic atrophy 1) and DRP-1 (Dynamin related protein 1). Our results showed the efficacy of KMP in hindering the toxicity induced by MPTP in D. melanogaster and suggest that the mitoprotective action of flavonoid may be boosting its anti-parkinsonism activity in the model. Besides, the study showed that wild-type strains of D. melanogaster proved to be reproducible in vivo model to mimic parkinsonian phenotypes through exposure to the neurotoxin MPTP.