Virtual platelet cross-matching as transfusion management for patients with immune platelet refractoriness.

BACKGROUND AND OBJECTIVES: This study describes the use of the Epvix platform for virtual cross-matching (VC) of human leucocyte antigen (HLA)-compatible platelets for patients with immune platelet refractoriness, and demonstrates effectiveness of the selected platelets. MATERIALS AND METHODS: A prospective cohort of haematological patients was evaluated from 2018 to 2022. HLA-typed donor bank profile was previously uploaded to the Epvix platform. Each patient's antibody reactivity panel (PRA) was included in the platform. Then, search, selection and VC were performed, and 24-h-corrected count increment (CCI) platelet transfusion was calculated (reference ≥2500). RESULTS: Six patients were included (four female, two male), with mean age of 61 years. HLA antibodies were detected as the cause of immunity for all patients, whereas four patients also had non-immune causes. High percentage of alloimmunization was detected in all studied patients (mean PRA: 85.7%). Thirty different donors were able to schedule and perform platelet donations. The mean 24-h CCI count was 9882. All platelet transfusions achieved a satisfactory CCI count except for two transfusion events. Presence of non-immune causes identified in these two cases could account for the unsatisfactory CCI. CONCLUSION: Epvix is a free application hosted on the Web and uses the HLAMatchmaker algorithm to generate histocompatibility reports. This study demonstrates the efficiency of VC performed by Epvix. However, physical cross-matching will still be necessary in some instances, as the platform does not support human platelet antigen polymorphism.

Synthetic antimicrobial peptides: activity against vancomycin-resistant Enterococcus faecalis and modulation of chloramphenicol antibacterial activity.

AIMS: Enterococcus faecalis (E. faecalis) is a leading cause of nosocomial infection and presents a wide spectrum of antibiotic resistance, being vancomycin-resistant Enterococcus (VRE) one of the most relevant. Synthetic antimicrobial peptides (SAMPs) are currently a promising option to overcome antimicrobial resistance. Thus, the purpose of this study was to assess the effect of eight SAMPs against vancomycin-resistant E. faecalis, as well as to investigate their mechanism of action and synergy with conventional antibiotics. METHODS AND RESULTS: Here, eight SAMPs, Mo-CBP3-PepI, Mo-CBP3-PepII, Mo-CBP3-PepIII, RcAlb-PepI, RcAlb-PepII, RcAlb-PepIII, PepGAT, and PepKAA, were tested for antibacterial activity in vitro against E. faecalis (ATCC® 51299) through broth microdilution. A maximum of 48% of E. faecalis growth inhibition was achieved by treatment with SAMPs alone. However, when these peptides were combined with the antibiotic chloramphenicol, assessed by checkerboard method, the inhibition increased to 55%-76% of inhibition, two to three-folds of increase if compared to the effects of the compounds alone. Microscopic analysis showed that E. faecalis cells treated with a combination of SAMPs and chloramphenicol resulted in bacterial membrane damage. The biofilm inhibition maximum was 22% for SAMPs alone, when combined with chloramphenicol, the maximum increased to 33%. CONCLUSIONS: SAMPs and their combination with chloramphenicol demonstrate antibacterial activity against E. faecalis, possibly by inducing bacterial membrane damage.

The synergic and addictive activity of biogenic silver nanoparticle associated with meropenem against carbapenem-resistant Acinetobacter baumannii.

AIMS: Antibiotic management of infections caused by Acinetobacter baumannii often fails due to antibiotic resistance (especially to carbapenems) and biofilm-forming strains. Thus, the objective here was to evaluate in vitro the antibacterial and antibiofilm activity of biogenic silver nanoparticle (Bio-AgNP) combined with meropenem, against multidrug-resistant isolates of A. baumannii. METHODS AND RESULTS: In this study, A. baumannii ATCC® 19606™ and four carbapenem-resistant A. baumannii (Ab) strains were used. The antibacterial activity of Bio-AgNP and meropenem was evaluated through broth microdilution. The effect of the Bio-AgNP association with meropenem was determined by the checkboard method. Also, the time-kill assay and the integrity of the bacterial cell membrane were evaluated. Furthermore, the antibiofilm activity of Bio-AgNP and meropenem alone and in combination was determined. Bio-AgNP has antibacterial activity with minimum inhibitory concentration (MIC) and minimum bactericidal concentration ranging from 0.46 to 1.87 µg ml-1. The combination of Bio-AgNP and meropenem showed a synergistic and additive effect against Ab strains, and Bio-AgNP was able to reduce the MIC of meropenem from 4- to 8-fold. Considering the time-kill of the cell, meropenem and Bio-AgNP when used in combination reduced bacterial load to undetectable levels within 10 min to 24 h after treatment. Protein leakage was observed in all treatments evaluated. When combined, meropenem/Bio-AgNP presents biofilm inhibition for Ab2 isolate and ATCC® 19606™, with 21% and 19%, and disrupts the biofilm from 22% to 50%, respectively. The increase in nonviable cells in the biofilm can be observed after treatment with Bio-AgNP and meropenem in carbapenem-resistant A. baumannii strains. CONCLUSIONS: The combination of Bio-AgNP with meropenem can be a therapeutic option in the treatment of infections caused by carbapenem-resistant A. baumannii.

Chimeric lipoproteins for leptospirosis vaccine: immunogenicity and protective potential.

Leptospirosis, a neglected zoonotic disease, is caused by pathogenic spirochetes belonging to the genus Leptospira and has one of the highest morbidity and mortality rates worldwide. Vaccination stands out as one of the most effective preventive measures for susceptible populations. Within the outer membrane of Leptospira spp., we find the LIC12287, LIC11711, and LIC13259 lipoproteins. These are of interest due to their surface location and potential immunogenicity. Thorough examination revealed the conservation of these proteins among pathogenic Leptospira spp.; we mapped the distribution of T- and B-cell epitopes along their sequences and assessed the 3D structures of each protein. This information aided in selecting immunodominant regions for the development of a chimeric protein. Through gene synthesis, we successfully constructed a chimeric protein, which was subsequently expressed, purified, and characterized. Hamsters were immunized with the chimeric lipoprotein, formulated with adjuvants aluminum hydroxide, EMULSIGEN®-D, Sigma Adjuvant System®, and Montanide™ ISA206VG. Another group was vaccinated with an inactivated Escherichia coli bacterin expressing the chimeric protein. Following vaccination, hamsters were challenged with a virulent L. interrogans strain. Our evaluation of the humoral immune response revealed the production of IgG antibodies, detectable 28 days after the second dose, in contrast to pre-immune samples and control groups. This demonstrates the potential of the chimeric protein to elicit a robust humoral immune response; however, no protection against challenge was achieved. While this study provides valuable insights into the subject, further research is warranted to identify protective antigens that could be utilized in the development of a leptospirosis vaccine. KEY POINTS: • Several T- and B-cell epitopes were identified in all the three proteins. • Four different adjuvants were used in vaccine formulations. • Immunization stimulated significant levels of IgG2/3 in vaccinated animals.

Antibacterial and Antileishmanial Activity of 1,4-Dihydropyridine Derivatives.

: We have synthesized twenty-three 1,4-dihydropyridine derivatives (1,4-DHPs) by using a microwave-assisted one-pot multicomponent Hantzsch reaction and evaluated their antibacterial activity against a representative panel of cariogenic bacteria and their in vitro antileishmanial activity against Leishmania (L.) amazonensis promastigotes and amastigotes. Thirteen compounds were moderately active against Streptococcus sanguinis, Streptococcus mitis, and Lactobacillus paracasei. Compound 22 (diethyl 4-(3-methoxy-4-hydroxyphenyl)-2,6-dimethyl-1,4-dihydropyridine-3,5-dicarboxylate) displayed moderate antibacterial activity against S. mitis and S. sanguinis, with a Minimum Inhibitory Concentration (MIC) of 500 µg/mL); compounds 8 (ethyl 2,7,7-trimethyl-4-(3-chlorophenyl)-5-oxo-1,4,5,6,7,8-hexahydroquinoline-3-carboxylate) and 10 (ethyl 2,7,7-trimethyl-4-(3-nitrophenyl)-5-oxo-1,4,5,6,7,8-hexahydroquinoline-3-carboxylate) were moderately active against S. sanguinis (MIC = 500 µg/mL) and very active against L. amazonensis promastigotes (IC50 = 43.08 and 34.28 µM, respectively). Among the eight 1,4-DHPs that were active (IC50 < 50 µM) against L. amazonensis promastigotes, compound 13 (ethyl 2,7,7-trimethyl-4-(3,4,5-trimethoxyphenyl)-5-oxo-1,4,5,6,7,8-hexahydroquinoline-3-carboxylate) was the most active (IC50 = 24.62 µM) and had a Selectivity Index (SI) higher than 4 compared to GM07492A cells. On the other hand, compound 9 (ethyl 2,7,7-trimethyl-4-(2-nitrophenyl)-5-oxo-1,4,5,6,7,8-hexahydroquinoline-3-carboxylate) was the most active against L. amazonensis amastigotes (IC50 = 16.27 µM and SI = 6.1) after 24 h of treatment. Based on our results, asymmetric 1,4-DHPs derived from dimedone exhibit antileishmanial potential.

Clinical Study of Photobiomodulation Effect at Postoperative Clefted Alveolar Ridge Bone Grafting.

This study aimed to evaluate photobiomodulation (PBM) adjunct effect on pain, swelling, and face temperature in individuals with oral cleft submitted to alveolar ridge bone grafting (ABG). The sample consisted of 46 patients with either incomplete or complete unilateral cleft lip and palate aged from 9 to 18 years, both sexes, who underwent ABG with chin donor area. Subjects were randomly divided into 3 groups: (1) Control (C): ABG (n=17); (2) PBM: ABG with PBM (n=14); (3) Simulated photobiomodulation (SPBM): ABG with SPBM (n=15). Aluminum Gallium Arsenide (GaAlAs) infrared laser was used at 808±10 nm and power of 100 mW±20%. GaAlAs infrared laser was applied on 14 points in the subject's face at 4J per point, at 2 times: Immediate postoperative period and 24 hours postoperative. The authors evaluated pain, swelling, and face temperature at 24 hours postoperative; at 7-day postoperative, the authors evaluated pain, medication use, and swelling decreasing perception. No statistically significant difference occurred between groups C, PBM, and SPBM. At 24 hours postoperative, pain values showed statistically significant difference (P=0.04) between group C and SPBM. PBM at the tested parameters did not reduce pain and swelling and did not significantly change face temperature of individuals submitted to ARBG.

Hormone replacement therapy did not alleviate temporomandibular joint inflammatory pain in ovariectomized rats.

This study had the aim of examining the relationships between variations in estrogen levels resulting from ovariectomy, and estrogen hormone replacement therapy (HRT) in rats subjected to an orofacial inflammatory pain model. Eighty adult female Wistar rats were initially divided into 2 groups: Sham or ovariectomy (OVX-D1). Seven days later (D7), the rats were subjected to an unilateral infiltration of Freund's Complete Adjuvant (CFA) or saline solution into the right temporomandibular joint (TMJ). Then, rats received 17ß-estradiol (28 µg/kg/day) or placebo for 21 days (D10-D31). Nociception was evaluated by the von Frey (VF) and the Hot Plate (HP) tests, and depressive-like behavior by the Forced Swimming (FS) test. On D32 all rats were euthanized and serum, hippocampus and brainstem were collected. The CFA groups presented a mechanical hyperalgesia until day 21 (p ≤ 0.05). No differences were observed among groups in the HP (p = 0.735), and in the immobility and swimming time of the FS (p = 0.800; p = 0.998, respectively). In the brainstem, there was a significant difference in the TNF-ɑ levels (p = 0.043), and a marginal significant difference in BDNF levels (p = 0.054), without differences among groups in the hippocampal BDNF and TNF-ɑ levels (p = 0.232; p = 0.081, respectively). In conclusion, the hormone replacement therapy did not alleviate orofacial pain in ovariectomized rats. However, there is a decrease in brainstem TNF-ɑ levels in the animals submitted to both models, which was partially reverted by HRT.

Facial Analysis of Patients with Unilateral or Bilateral Cleft Lip and Palate Using 3D Stereophotogrammetry.

OBJECTIVES: To evaluate the quantitative values of linear and angular facial anthropometrics in patients with unilateral cleft lip and palate (UCLP) and compare them with those of patients with bilateral cleft lip and palate (BCLP) using three-dimensional (3D) facial imaging. DESIGN: Retrospective, observational, and cross-sectional study. SETTING: Hospital for Rehabilitation of Craniofacial Anomalies/USP (HRAC/USP). PATIENTS/ PARTICIPANTS: In total, 61 non-syndromic patients with CLP who underwent multidisciplinary treatment and rehabilitated with a prosthesis were enrolled and divided into those with UCLP (G1; n = 31) and those with BCLP (G2; n = 30). INTERVENTION: Facial images were captured using a 3D camera after landmarks were marked on each patient's face. The software evaluated linear and angular parameters. Statistical tests were applied. Significance was determined as P < 0.05. MAIN OUTCOME MEASUREMENTS: Overall, 22 linear and 13 angular measurements were evaluated. RESULTS: The nasal length (P = 0.08), middle third of the face (P = 0.06), base nose width (P < 0.001), nasal root width (P < 0.001), nasal tip angle (P = 0.018), philtrum width (P < 0.001), lower face width (P = 0.039) and midfacial depth (P = 0.040) were significantly higher in G2; the upper cutaneous lip height was significantly higher in G1. Sexual dimorphism was observed except for linear measurements (linear distance between the labiale superius and labiale inferius landmarks, nasal root width, and upper cutaneous lip length) and angular measurements. CONCLUSIONS: G2 had a greater length and width of nose and nasal root, nasal tip angle, philtrum width, and lower face width, midfacial depth, and midface third than G1. These findings also revealed the presence of sexual dimorphism.

Saltiness perception in gel-based food systems (gels and emulsion-filled gels).

Reducing salt in food without compromising its quality is a huge challenge. Some review articles have been recently published on saltiness perception in some colloidal systems such as emulsions. However, no published reviews are available on saltiness perceptions of gel-based matrices, even though salt release and perception in these systems have been extensively studied. This article reviews the recent advances in salt perception in gel-based systems and provides a detailed analysis of the main factors affecting salt release. Strategies to enhance saltiness perception in gels and emulsion-filled gels are also reviewed. Saltiness perception can be improved through addition of biopolymers (proteins and polysaccharides) due to their ability to modulate texture and/or to adhere to or penetrate through the mucosal membrane on the tongue to prolong sodium retention. The composition of the product and the distribution of salt within the matrix are the two main factors affecting the perception of salty taste. Food structure re-design can lead to control the level of interaction between the salt and other components and change the structure, which in turn affects the mobility and release of the salt. The change of ingredients/matrix can affect the texture of the product, highlighting the importance of sensory evaluation.

What Are the Effects on Palate of Early Lip Surgery in Children With Cleft Lip and Palate? Cross-Sectional Evaluation From 5-Year-Old.

This study aimed to evaluate the postsurgical effects from 5 years on the palate after surgical repair of the lip at 3 or 9 months of age in children with cleft lip and palate. Eighty-four digitized dental impressions were divided into the following groups: group 1 (G1): lip surgery at 3 months of life; group 2 (G2): lip surgery at 9 months of life; group 3 (G3): without orofacial cleft. Five angular (C'IC, ICM, IC'M', CMM', and C'M'M) and 3 linear parameters (C-C', c-c', and M-M') were evaluated. Statistical analysis was applied with α=5%. Intraclass Correlation Coefficient was significantly smaller in G1 than in G3 ( P =0.005), while IC'M' was significantly smaller in G3 than in G1 ( P <0.001). C'M'M was significantly smaller in G1 than in G2 and G3 ( P <0.001). The distances C-C' and c-c' were significantly smaller in G1 than in G2 and G3 ( P <0.001). There was a statistically significant difference in both G1 and G2 ( P <0.001, in all) in the analysis of palatal symmetry. Linear regression analysis showed that the, 11.2% of outcomes determined by c-c' distance can be explained by the age of lip repair ( P =0.013). In conclusion, lip surgery at 3 months of life showed a tendency toward more restriction in 5-year postsurgery palate development. The age of cheiloplasty is one of the factors that can influence palatal development; however, other factors may be associated and should be studied.

Therapeutic efficacy of enrofloxacin in treatment of Francisella orientalis infections in juvenile Nile tilapia (Oreochromis niloticus L.).

Outbreaks of infections by Francisella orientalis represent one of the main obstacles to Nile tilapia (Oreochromis niloticus L.) farming. It is responsible for acute mortality in fingerlings and juveniles. The main control measure available is oral antibiotic therapy. This study compared the therapeutic efficacy of the antibiotics enrofloxacin and oxytetracycline, the most commonly used antimicrobial, against francisellosis in juvenile Nile tilapia (O. niloticus). Fish were challenged with a virulent isolate of F. orientalis and treated with medicated feed containing one of two doses of oxytetracycline (100 or 300 mg/kg of live weight (LW)) or 10 mg/kg of LW of enrofloxacin. The positive and negative control groups received feed without antibiotics; the negative control group was unchallenged. The results showed that enrofloxacin at a dose of 10 mg/kg of LW is effective against francisellosis in juvenile Nile tilapia (O. niloticus). Treatment with oxytetracycline did not eliminate the pathogen from the infected host, and the surviving fish became carriers. Enrofloxacin was able to cure the fish of infection with F. orientalis. This study suggests that enrofloxacin is a better option for treating francisellosis in Nile tilapia (O. niloticus L.). It controls mortality and avoids the carrier state in the fish, thus reducing the possibility of recurrence in the affected batches.

Customized Tray for Impression Taking in Children With Cleft Lip and Palate.

The assessment of rehabilitation outcomes requires a patient documentation protocol, including records obtained at standardized ages, to compare different types of surgeries, their effects, as well as between different rehabilitation centers. The aim of this paper was to present proper trays for babies with different types of cleft lip and palate, which are used in the outpatient routine at Hospital of Rehabilitation of Craniofacial Anomalies/USP (HRAC/USP). The customized trays are made with self-curing acrylic resin. The tray must have suitable depth to copy the buccal sulcus, and wax is usually applied to contour the tray edge, and the adjustment of the tray to the fornix, making the tray specific for each child. The impression precludes the utilization of dental casts for diagnosis, treatment plan, and research measurements. In the clinical practice at HRAC-USP, it was observed that customized trays increased the quality of impression, accurately reproducing anatomical features of dental arches of babies with oral clefts.

Arch Symmetry in Patients Without and With Cleft Lip and Palate After Orthodontic/Rehabilitative Treatment-A Stereophotogrammetry Study.

To evaluate and compare the dental arch symmetry of individuals with and without cleft lip and palate after orthodontic/rehabilitation treatment.Cross-sectional study.Tertiary cleft center in Brazil.Fifty-five participants aged between 18 and 30 years were divided into 3 groups according to treatment. Patients that received either a fixed partial denture (FPD) or implant-supported crown (ISC) in the cleft area or only orthodontic treatment, noncleft patients (NC).An analysis was performed using digitized dental casts scanned by laser and software. The following linear measurements were evaluated: incisor-canine; canine-molar; incisor-molar; surface and volume of the palatal region.Three-way ANOVA was used to compare the study factors: group (FPD/ISC/NC) and side (right/left) followed by the Tukey test to verify their interaction (α = .05).The results showed statistically significant differences among groups for the maxillary linear measurements canine-molar and incisor-molar, but not for incisor-canine. No statistically significant differences were found regarding the side for the maxillary measurements, while the factor interaction showed similarity only for incisor-canine. The mandibular measurements showed no statistical differences among groups, sides, or factor interactions. In surface and volume, all values in patients with cleft presented lesser than in without cleft patients.Regardless of the rehabilitation, arch symmetry can be achieved in the incisor-canine dimension in the cleft area.

Nursing Interventions for Patients With Neurogenic Bowel Dysfunction Arising From Complete Spinal Cord Injury: A Scoping Review.

PURPOSE: We mapped key concepts and identified 4 fundamental nursing interventions for patients with neurogenic bowel dysfunction due to complete spinal cord injury (SCI). METHODS: A scoping review was conducted according to the recommendations of the Joanna Briggs Institute. SEARCH STRATEGY: Searches were performed in PubMed, LILACS, CINAHL, COCHRANE, and SCOPUS electronic databases. We use searched the gray literature using the Google Scholar search engine. We formulated a question to guide the search, based on the participants, concept, and context format: "What are the key manual nursing interventions performed in patients with neurogenic bowel dysfunction resulting from complete spinal cord injury?" We included nursing intervention strategies that may be performed by health professionals, patients, or caregivers. Two reviewers independently participated in the selection; disagreements were resolved by a third reviewer and 5 experts. FINDINGS: Thirteen studies conducted between 1998 and 2019 were selected; 5 were randomized clinical trials. Four main interventions were identified for conservative management of neurogenic bowel dysfunction in patients with complete SCI. They were digital-anal stimulation, manual extraction of feces, abdominal massage, and strategies used to stimulate the gastrocolic reflex. CONCLUSIONS: Research suggests that each of these interventions, administered alone or in combination, supports bowel evacuation in patients with a complete SCI. Each of these interventions may be performed by a nurse, and taught to the patient and/or lay caregiver. IMPLICATIONS FOR PRACTICE: An individualized bowel management program for patients with neurogenic bowel dysfunction due to SCI is necessary to ensure regular bowel evacuation, preserve fecal continence, and support dignity and health-related quality of life. The conservative interventions identified in this scoping review should be incorporated in protocols or guidelines for management of neurogenic bowel dysfunction in this vulnerable population.

Ectopic expression of an expansin-like B gene from wild Arachis enhances tolerance to both abiotic and biotic stresses.

Plant expansins are structural cell wall-loosening proteins implicated in several developmental processes and responses to environmental constraints and pathogen infection. To date, there is limited information about the biological function of expansins-like B (EXLBs), one of the smallest and less-studied subfamilies of plant expansins. In the present study, we conducted a functional analysis of the wild Arachis AdEXLB8 gene in transgenic tobacco (Nicotiana tabacum) plants to clarify its putative role in mediating defense responses to abiotic and biotic stresses. First, its cell wall localization was confirmed in plants expressing an AdEXLB8:eGFP fusion protein, while nanomechanical assays indicated cell wall reorganization and reassembly due to AdEXLB8 overexpression without compromising the phenotype. We further demonstrated that AdEXLB8 increased tolerance not only to isolated abiotic (drought) and biotic (Sclerotinia sclerotiorum and Meloidogyne incognita) stresses but also to their combination. The jasmonate and abscisic acid signaling pathways were clearly favored in transgenic plants, showing an activated antioxidative defense system. In addition to modifications in the biomechanical properties of the cell wall, we propose that AdEXLB8 overexpression interferes with phytohormone dynamics leading to a defense primed state, which culminates in plant defense responses against isolated and combined abiotic and biotic stresses.

Development of an inactivated whole cell vaccine through immersion for immunoprophylaxis of Francisella orientalis infections in Nile tilapia (Oreochromis niloticus L.) fingerlings and juveniles.

Francisella orientalis infections, known as francisellosis, are one of the most important diseases affecting the production of Nile tilapia, causing high mortality rates in the most susceptible fish stages: fingerlings and juveniles. Antibiotic therapy is the method of choice for treating the disease, as there are no commercially available vaccines. In this study, we developed an inactivated whole-cell vaccine using an isolate of F. orientalis in combination with the aqueous adjuvant Montanide IMS 1312 VG, which was administered to Nile tilapia through immersion. Two immunization trials (1 and 2) were conducted with fish at the fingerling and juvenile stages. For each trial, five different experimental groups were established: a complete vaccine (bacterin in combination with aqueous adjuvant), bacterin, aqueous adjuvant, and positive and negative controls. Thirty days after vaccination, an experimental challenge was performed through intraperitoneal injection of the same F. orientalis isolate. As a result, the vaccinated fingerlings were the only group in which mortality and progression of clinical signs of francisellosis were statistically significantly reduced, although relative percentage of survival (RPS) was low at 50%. In the juvenile group, RPS was higher at 63%, but not statistically significant. Nevertheless, an RPS of only 50% is acceptable for using vaccines in the field. The bacterin and adjuvant treatments alone were not effective, showing an RPS of 37% and 0%, respectively. Post-vaccination mortality was observed in the group exposed only to the adjuvant, which may indicate excessive immune stimulation at this stage. Interestingly, the immune response elicited by the vaccine was unable to eliminate the pathogen from the host; therefore, the surviving animals became carriers. Although the immune response elicited by the vaccine was unable to eliminate the pathogen from the host, this vaccine formulation could be a viable alternative for use in the field and serve as another means of controlling the mortality caused by the pathogen. Our study provides the first report of vaccination, using immersion, against francisellosis at the most susceptible stages of farmed Nile tilapia. Future studies should address the efficiency of immersion vaccines under field conditions.

TonB-dependent receptor epitopes expressed in M. bovis BCG induced significant protection in the hamster model of leptospirosis.

Leptospirosis is an emerging infectious disease caused by pathogenic Leptospira spp. A universal vaccine against leptospirosis is likely to require highly conserved epitopes from pathogenic leptospires that are exposed on the bacterial surface and that generate a protective and sterilizing immune response. Our group recently identified several genes predicted to encode TonB-dependent receptors (TBDR) in Leptospira interrogans using a reverse vaccinology approach. Three leptospiral TBDRs were previously described and partially characterized as ferric-citrate, hemin, and cobalamin transporters. In the current study, we designed a fusion protein composed of predicted surface-exposed epitopes from three conserved leptospiral TBDRs. Based on their three-dimensional structural models and the prediction of immunogenic regions, nine putative surface-exposed fragments were selected to compose a recombinant chimeric protein. A Mycobacterium bovis BCG strain expressing this chimeric antigen encoded in the pUP500/PpAN mycobacterial expression vector was used to immunize Syrian hamsters. All animals (20/20) vaccinated with recombinant BCG survived infection with an endpoint dose of L. interrogans (p < 0.001). No animal survived in the negative control group. Immunization with our recombinant BCG elicited a humoral immune response against leptospiral TBDRs, as demonstrated by ELISA and immunoblot. No leptospiral DNA was detected by lipL32 qPCR in the kidneys of vaccinated hamsters. Similarly, no growth was observed in macerated kidney cultures from the same animals, suggesting the induction of a sterilizing immune response. Design of new vaccine antigens based on the structure of outer membrane proteins is a promising approach to overcome the impact of leptospirosis by vaccination. KEY POINTS: • Predicted surface-exposed epitopes were identified in three leptospiral TBDRs. • An M. bovis BCG strain expressing a chimeric protein (rTBDRchi) was constructed. • Hamsters vaccinated with rBCG:TBDRchi were protected from lethal leptospirosis.

Digital surveillance: The interests in toothache-related information after the outbreak of COVID-19.

OBJECTIVES: The aim of this ecological study was to analyze the relationship of social restriction measures and people's interest in searching toothache-related information on the Internet. MATERIALS AND METHODS: The following indicators Stringency Index (SI), Years Lived with Disability (YLDs) for untreated caries in permanent teeth, Google market share, Internet penetration, and relative search volume (RSV) for the Topic "Toothache" in Google Trends were determined for 21 countries with available data, from April 2018 to May 2020. Statistical analysis was performed using Mann-Whitney U test, t-Student test, Pearson's Chi-square test, and Spearman and Pearson's cross-correlation tests (α = 0.05). RESULTS: Relative search volume values increased significantly after restriction measures in all countries, except for Japan. Higher RSV values were found among 11 countries that presented a cross-correlation between RSV and SI with r≥│0.6│. The most common queries were linked to the treatment and self-resolution of toothache, with a sudden increase of searches that combined the symptom with COVID-19. CONCLUSIONS: The interests in toothache-related digital information were associated with social restriction measures. These results allow the identification of dental demands of distinct populations, contributing to the planning of specific public health policies during and after the pandemic period.

The Brazilian Longitudinal Study of Adult Health (ELSA-Brasil): Factors Related to Smoking Cessation.

BACKGROUND: Smoking cessation is not an easy accomplishment. However, the benefits are several for those who do it, such as cardiovascular risk reduction 1 year after quitting smoking. This study aimed to verify the factors related to smoking cessation in civil servants of The Brazilian Longitudinal Study of Adult Health (ELSA-Brasil). METHODS: This study had a longitudinal design using data from a prospective cohort of civil servants (ELSA-Brasil). Our variable of interest was smoking cessation. The relationship between socio-demographic characteristics, job stress, health-related variables, legislation, and smoking cessation was analyzed by Cox proportional hazard models. The analyses were stratified by gender. Second-hand smoke exposure, age, education, excessive alcohol consumption, common mental disorder, and smoking control law were the variables considered in the final model. RESULTS: Information of 2020 women and 2429 men was analyzed. Individuals without second-hand smoke exposure, with up to 49 years of age, with higher education, without excessive alcohol consumption, without common mental disorders, and who initiated smoking in 1989 or after the smoking control law had a higher risk of stopping smoking. The risks magnitudes were higher for women. CONCLUSIONS: Our study reinforces the necessity of alcohol consumption regulation, the relevance of Public Health Policies, and the need for more smoking cessation measures focused on men, on people with mental disorders, alcoholism, and older adults. Also, our results did not show significant risks regarding the psychosocial working environment.

Cost analysis of acute care resource utilization among individuals with sickle cell disease in a middle-income country.

BACKGROUND: The costs associated with the treatment of sickle cell disease (SCD) are understudied in low and middle-income countries (LMIC). We evaluated the cost of treating SCD-related acute complications and the potential cost-savings of hydroxyurea in a specialized hematology center in Brazil. METHODS: The costs (US dollars) of emergency department (ED) and hospitalizations from SCD-related complications between 01.01.2018 and 06.30.2018 were ascertained using absorption and micro-costing approaches. The reasons for acute hospital visits were grouped as: 1) vaso-occlusive (VOC) pain, 2) infection, 3) anemia exacerbation, and 4) chronic organ damage complications. Hydroxyurea adherence was estimated by medication possession ratio (MPR) during the study period. RESULTS: In total, 1144 patients, median age 17 years (range 0-70), 903 (78.9%) with HbSS/HbSß0-thalassemia, 441 (38.5%) prescribed hydroxyurea, visited the ED, of whom 381 (33%) were admitted. VOC accounted for 64% of all ED visits and 60% of all admissions. Anemia exacerbation was the most expensive reason for ED visit ($321.87/visit), while chronic organ damage carried the highest admission cost ($2176.40/visit). Compared with other genotypes, individuals with HbSS/HbSß0-thalassemia were admitted more often (79% versus 21%, p < 0.0001), and their admission costs were higher ($1677.18 versus $1224.47/visit, p = 0.0001). Antibiotics and analgesics accounted for 43% and 42% of the total ED costs, respectively, while housing accounted for 46% of the total admission costs. Costs of ED visits not resulting in admissions were lower among HbSS/HbSß0-thalassemia individuals with hydroxyurea MPR ≥65% compared with visits by patients with MPR <65% ($98.16/visit versus $182.46/visit, p = 0.0007). No difference in admission costs were observed relative to hydroxyurea use. DISCUSSION: In a LMIC hematology-specialized center, VOCs accounted for most acute visits from patients with SCD, but costs were highest due to anemia exacerbation. Analgesics, antibiotics, and housing drove most expenses. Hydroxyurea may reduce ED costs among individuals with HbSS/HbSß0-thalassemia but is dependent on adherence level.