Development and validation of an LC-ESI-QTOF-MS method to measure cefepime in the plasma and peritoneal fluid of rats using microdialysis: Application in a pilot pharmacokinetic study.

Cefepime (CEF) is a cephalosporin and can be administered in secondary peritonitis together with metronidazole to treat sepsis. This study aimed to develop and validate an LC-ESI-QTOF-MS method for the quantification of cefepime in the plasma and peritoneal microdialysate of healthy Wistar rats. Chromatographic separation was performed using a CLC-ODS C18 column (250 × 4.6 mm), a C18 pre-column (4 mm, 5 µm) and isocratic elution. Gallic acid was used as the internal standard. The mobile phase consisted of (A) ultrapure water (pH adjusted to 3.5) and (B) acetonitrile (80:20, v/v) at 0.8 ml/min. Quantification was performed using a mass spectrometer with electrospray ionization in positive mode to monitor ions with m/z 481.1322 (CEF) and m/z 171.0288 (IS). The method was validated for selectivity, precision, accuracy, linearity, stability, lower limit of quantification, carryover, recovery and matrix effect. Calibration was done in the ranges 1-40 and 1-100 µg/ml for the peritoneal microdialysate and plasma, respectively. Plasma extraction recovery ranged from 93.9 to 99.9%. The technique was validated and successfully applied in a pilot pharmacokinetic study for estimating the free concentration of CEF in the peritoneal microdialysate of rats for the first time.

Four almost unexplored species of Brazilian Connarus (Connaraceae): Chemical composition by ESI-QTof-MS/MS-GNPS and a pharmacologic potential.

INTRODUCTION: Species of Connaraceae are globally used in traditional medicines. However, several of these have not been studied regarding their chemical composition, and some are even at risk of extinction without proper studies. Therefore, the chemical composition and pharmacological potential of Connarus blanchetii Planch., Connarus nodosus Baker, Connarus regnellii G. Schellenb., and Connarus suberosus Planch., which were previously unknown, were analyzed. OBJECTIVE: This work aims to investigate the pharmacological potential of these four Connarus species. The chemical composition of different extracts was determined by high-resolution mass spectrometry (HRMS), with subsequent analysis by the GNPS platform and competitive fragmentation modeling (CFM). MATERIALS AND METHODS: Leaf extracts (C. blanchetii, C. nodosus, C. regnellii, and C. suberosus) and bark extracts (C. regnellii and C. suberosus) were obtained by decoction, infusion, and maceration. LC/HRMS data were submitted to the GNPS platform and evaluated using CFM in order to confirm the structures. RESULTS: The HRMS-GNPS/CFM analysis indicated the presence of 23 compounds that were mainly identified as phenolic derivatives from quercetin and myricetin, of which 21 are unedited in the Connarus genus. Thus, from the analyses performed, we can identify different compounds with pharmacological potential, as well as the most suitable forms of extraction. CONCLUSION: Using HRMS-GNPS/CFM, 21 unpublished compounds were identified in the studied species. Therefore, our combination of data analysis techniques can be used to determine their chemical composition.

Shockwave Therapy Plus Eccentric Exercises Versus Isolated Eccentric Exercises for Achilles Insertional Tendinopathy: A Double-Blinded Randomized Clinical Trial.

BACKGROUND: There remains a lack of consensus regarding the treatment of Achilles insertional tendinopathy. The condition is typically treated with eccentric exercises despite the absence of satisfactory and sustained results. Shockwave therapy was presented as an alternative, but there is a paucity of literature, with good outcomes, supporting its use. The purpose of the present single-center, double-blinded, placebo-controlled, randomized trial was to determine if the use of shockwave therapy in combination with eccentric exercises improves pain and function in patients with Achilles insertional tendinopathy. METHODS: A total of 119 patients with Achilles insertional tendinopathy were evaluated and enrolled in the study from February 2017 to February 2019. Patients were allocated to 1 of 2 treatment groups, eccentric exercises with extracorporeal shockwave therapy (SWT group) and eccentric exercises with sham shockwave therapy (control group). Three sessions of radial shockwaves (or sham treatment) were performed every 2 weeks and eccentric exercises were undertaken for 3 months. The primary outcome was the Victorian Institute of Sport Assessment-Achilles questionnaire (VISA-A) at 24 weeks. Secondary outcomes included the visual analogue scale, algometry, the Foot and Ankle Outcome Score, and the 12-Item Short Form Health Survey. RESULTS: Both groups showed significant improvement during the study period; however, there were no between-group differences in any of the outcomes (all p >0.05). At the 24-week evaluation, the SWT group exhibited a mean VISA-A of 63.2 (95% confidence interval, 8.0) compared with 62.3 (95% confidence interval, 6.9) in the control group (p = 0.876). There was a higher rate of failure (38.3%) but a lower rate of recurrence (17.0%) in the SWT group compared with the control group (11.5% and 34.6%, respectively; p = 0.002 and p = 0.047). There were no complications reported for either group. CONCLUSIONS: Extracorporeal shockwave therapy does not potentiate the effects of eccentric strengthening in the management of Achilles insertional tendinopathy. LEVEL OF EVIDENCE: Therapeutic Level I. See Instructions for Authors for a complete description of levels of evidence.

Synthesis, chemical characterization and DNA interaction study of new diclofenac and ibuprofen zinc (II)-nicotinamide ternary complexes as cyclooxygenase inhibitor prototypes.

In the search for new drugs, strategies such as bioisosterism have been evidenced, in which the modification of molecules is already known to be active. Thus, metal complexes of known drugs have been highlighted, with examples of significant improvements in therapeutic efficacy. In this way, this work aimed at the synthesis of new zinc complexes with nonsteroidal anti-inflammatory drugs (NSAIDs), as well as the chemical characterization and the previous toxicity by cytotoxicity with Artemia salina, and evaluating the ability of these compounds to interact with DNA. As result, two new zinc II ternary complexes containing the NSAIDs diclofenac (Diclof) and ibuprofen (Ibup) and nicotinamide neutral linker (Nic) were obtained by the two-step solvent metal-ligand complexation method. Molecular structures were determined by NMR, FTIR, HR-MS, UV-Vis and X-ray diffraction, which demonstrated that both complexes are binuclear systems of general formula [Zn2(R-COO-)4(Nic)2]. Plasmidial DNA breakdown capacities were evaluated by producing single and double breaks (DNA FII and FIII) from plasmid incubation with complex solutions in the concentration range 0 to 400 µmol·L-1 in experiments with the presence and absence of light. Both experiments did not show significant differences (P ≤ 0.05) in induced DNA cleavage activity between the maximum study concentrations (400 µmol·L-1) and the negative controls for both complexes. The types of complex 1 and 2 interactions with the secondary DNA structure were determined by titrating a CT-DNA solution with complex solutions and monitored by circular dichroism spectrometry. The results showed that both complexes interact with the grooves of the secondary structure of CT-DNA by electrostatic attraction, but without evidence of alteration in the primary structure. Acute toxicity tests against Artemia salina showed that both complexes did not produce lethality >10% of the population up to a maximum concentration of 1200 µg·mL-1 within an exposure interval of 24 h. Thus, two new compounds were synthesized, characterized and had their previous toxicities determined. These compounds are promising new drugs, with the next step being evaluations of their activity.

Shockwave Therapy Associated With Eccentric Strengthening for Achilles Insertional Tendinopathy: A Prospective Study.

Background. The usual initial treatment for insertional Achilles tendinopathy is nonsurgical. Yet there is no standard conservative treatment for Achilles insertional tendinopathy. Shockwave therapy (SWT) has become a reliable option for the management of this illness over the past years. The aim of this study is to report the effectiveness of low-energy SWT associated with an eccentric strengthening protocol in 19 consecutive patients. Methods. This is a prospective study with 19 patients aged between 26 and 72 years diagnosed with insertional Achilles tendinopathy. The protocol consisted of SWT associated with eccentric exercises for 12 weeks. All patients were evaluated on the first day and after 24 weeks (final follow-up) with the Victorian Institute of Sports Assessment-Achilles (VISA-A) score, visual analogue scale (VAS), American Orthopaedic Foot and Ankle Society (AOFAS) questionnaire, and by algometry. At the last follow-up, patients were also assessed for adherence to the protocol, complications and final outcome (in their perception as success or fail). Results and Conclusion. Fifteen (79%) patients were fully adherent to the Alfredson protocol, and 13 (68%) patients considered the treatment protocol successful. At the last evaluation, patients demanded higher pressure on calcaneus to trigger pain (algometry 1), reported less pain when the algometer was applied with 3 kg (algometry 2), had less global pain (VAS), and had higher AOFAS and VISA-A scores. This study evidences that eccentric loading associated with SWT can dramatically improve patients' symptoms. We can conclude that eccentric loading associated with SWT is an effective treatment for Achilles insertional tendinopathy. Levels of Evidence: Therapeutic, Level III: Prospective cohort.

How do seasonality and host traits influence the distribution patterns of parasites on juveniles and adults of Columba livia?

Parasites may influence host fitness and consequently exert a selective pressure on distinct phenotypes of the host population. This pressure can result in an evolutionary response, maintaining only individuals with certain traits in the population. The present study was aimed at identifying the morphological characteristics of juveniles and adults of Columba livia that may influence the distribution patterns of lice, Pseudolynchia canariensis and Haemoproteus columbae and how the populations of these parasites vary throughout the seasons of the year. Between July 2012 and July 2014, 377 specimens of C. livia were captured. We observed a significant increase in the mean intensities of infestation by pigeon flies and lice, as well as in species richness of ectoparasites during the warmest seasons, suggesting a reproductive synchrony between ectoparasites and host species. Bill length, body mass, and body length did not affect the infestation levels of ectoparasites on adults and juveniles of C. livia with three distinct plumage colors. In juveniles, plumage color affected only the mean intensity of infestation by lice, with Spread individuals as the most infested. This indicates that melanin in feathers was not an effective barrier against ectoparasites.

Scorpionism by Tityus silvestris in eastern Brazilian Amazon.

BACKGROUND: Scorpionism is a serious public health problem in Brazil. Although cases of envenomation by scorpions are frequent in Brazil, Tityus silvestris - found throughout the Amazon region - is considered of minor medical significance and with only a few descriptions in the literature. This article aims to describe for the first time the epidemiological characteristics and clinical manifestations of scorpion stings by T. silvestris that occurred in eastern Brazilian Amazon. METHODS: A prospective and observational study was carried out on 13 confirmed cases of T. silvestris envenomation registered from 2007 to 2011 in the cities of Belém and Ananindeua, Pará state, Brazil. RESULTS: The stings occurred mainly during daytime, at domiciliary environment, and the scorpions were found in clothing, fruits or vegetables. Envenomation was more frequent in the age group between 21 and 30 years old, upper limbs were more affected and medical aid was usually provided within two hours. Men and women were equally affected. Regarding severity, ten patients were classified as Class I and three patients as Class II according to the Scorpion Consensus Expert Group. Local manifestations were present in all patients, being pain the most common symptom. Mild systemic manifestations including nausea, vomiting, somnolence, malaise and prostration were observed in three victims. Symptomatic treatment of pain was offered to all patients, and only one received specific antivenom. All victims had a favorable outcome. CONCLUSIONS: To the best of our knowledge, this study is the first to report the systemic symptomatology of envenomation by T. silvestris in the Brazilian Amazon, highlighting the medical relevance of the species in this region. Further research on the venom and clinical manifestations of envenomation by T. silvestris should be conducted in order to verify the relevance of this species to public health.

Composition of the essential oils from leaves of Piper lepturum Kunth (C.DC.) var. lepturum and Piper lepturum var. angustifolium (Miq.) Yunck. from Brasil / Composición de los aceites esenciales de hojas de Piper lepturum Kunth (C.DC.) var. lepturum y Pip

The essential oils of Brazilian Piper lepturum var. lepturum and Piper lepturum var. angustifolium (Piperaceae) were obtained by hydrodistillation and analyzed by flame-detector gas chromatography (GC) and gas chromatography coupled to mass spectrometry (GC/MS). According to GC and GC/MS analysis, the essential oils are mostly composed by sesquiterpenes hydrocarbons. beta-Guaiene (29.96 percent) was the principal component in the essential oil of P. lepturum var. lepturum and beta-Bisabolene (17.72 percent) was the principal components in the essential oil of P. lepturum var. angustifolium.

Los aceites esenciales de las especies brasileñas Piper lepturum var. lepturum y Piper lepturum var. angustifolium fueron obtenidos por hidrodestilación y analizados utilizando cromatografía gas líquido con detector de ionización de llama (CG) y cromatografía gas líquido acoplada a un detector de masas (CG/EM). De acuerdo con los análisis de CG y CG/EM, los aceites esenciales muestran como componente principal beta-Guaieno (29,96 por ciento) en el aceite esencial de P. lepturum var. lepturum y beta-Bisaboleno (17,71 por ciento) en el aceite esencial de P. lepturum var. angustifolium.

Effects of shock wave therapy on glycosaminoglycan expression during bone healing.

BACKGROUND: Several cases of delayed bone consolidation have been treated with extracorporeal shock wave therapy (ESWT) to improve bone healing and a key role of the extracellular matrix glycosaminoglycans in osteogenesis has been suggested. OBJECTIVE: In this study, we aimed to identify and quantify the amount of sulfated glycosaminoglycans (GAG) and hyaluronic acid (HA) within rat femurs following bone drilling and treatment with shock waves. METHODS: To identify and quantify the sulfated glycosaminoglycans (GAG) and hyaluronic acid (HA) within rat femurs following bone drilling and ESWT, 50 male Wistar rats were evaluated. The animals were divided into two groups, both of which were subjected to bone drilling. One of the groups was treated with ESWT. The rats were sacrificed on the 3rd, 7th, 14th, 21st, and 28th day. GAG presence was analyzed by agarose gel electrophoresis with subsequent densitometry and ELISA. RESULTS AND DISCUSSION: The content of sulfated GAGs increased significantly from the 3rd to the 28th day (p = 0.002). Chondroitin sulfate was expressed more highly than the other GAGs. HA content increased significantly at the 3rd day in animals treated with ESWT compared to the control group (p = 0.003). CONCLUSION: ESWT stimulates of sulfated glycosaminoglycans during bone healing and enhanced early expression of HA compared to the control group.

Characterization of Tn3000, a Transposon Responsible for blaNDM-1 Dissemination among Enterobacteriaceae in Brazil, Nepal, Morocco, and India.

In Enterobacteriaceae, the blaNDM genes have been found in many different genetic contexts, and a wide diversity of plasmid scaffolds bearing those genes has been found. In August 2013, we identified NDM-1-producing Escherichia coli and Enterobacter hormaechei strains from a single rectal swab sample from a patient hospitalized in Rio de Janeiro, Brazil, who had no history of travel abroad. Complete DNA sequencing using the Illumina platform and annotation of the two plasmids harboring the blaNDM-1 gene, one from each strain, showed that they belonged to incompatibility groups IncFIIK and IncX3 and harbored a novel transposon named Tn3000. Similar genetic structures have been identified among other isolates in Brazil but also on plasmids from other continents. Our findings suggest that the blaNDM-1 gene may be transmitted by Tn3000 in different parts of the world.

EXTRACORPOREAL SHOCKWAVE TERAPY TO TREAT CHRONIC MUSCLE INJURY.

OBJECTIVE: To evaluate the low energy extracorporeal shock waves therapy (ESWT) associated with physical therapy in the treatment of chronic muscle injuries classified as grades 2 and 3 in the lower limbs of amateur athletes. METHODS: Eight athletes presenting with lower limb muscle injury for more than three weeks were treated with physiotherapy and ESWT. We evaluated the following parameters during treatment: palpable gap, muscle strength, pain, and Tegner score, as well as ultrasound image features and the ability to return to sports practice. RESULTS: The average time of the first evaluation of the injury was 8.75 weeks. All patients presented muscle strength grade V after eight weeks. The pain score evolved from 5.75 to 0.5 points of the visual analogue scale (VAS), at the end of the treatment. The Tegner score after treatment was six points on average. Patients returned to sports practice after 8.14 weeks. CONCLUSION: ESWT associated with physical therapy proved to be effective to treat long-term muscle injury, with good performance and the ability to return to sport practice for all patients. Level of Evidence IV, Case Series, Prospective Study.

Adverse events following yellow fever immunization: Report and analysis of 67 neurological cases in Brazil.

Neurological adverse events following administration of the 17DD substrain of yellow fever vaccine (YEL-AND) in the Brazilian population are described and analyzed. Based on information obtained from the National Immunization Program through passive surveillance or intensified passive surveillance, from 2007 to 2012, descriptive analysis, national and regional rates of YFV associated neurotropic, neurological autoimmune disease, and reporting rate ratios with their respective 95% confidence intervals were calculated for first time vaccinees stratified on age and year. Sixty-seven neurological cases were found, with the highest rate of neurological adverse events in the age group from 5 to 9 years (2.66 per 100,000 vaccine doses in Rio Grande do Sul state, and 0.83 per 100,000 doses in national analysis). Two cases had a combination of neurotropic and autoimmune features. This is the largest sample of YEL-AND already analyzed. Rates are similar to other recent studies, but on this study the age group from 5 to 9 years of age had the highest risk. As neurological adverse events have in general a good prognosis, they should not contraindicate the use of yellow fever vaccine in face of risk of infection by yellow fever virus.

Community of arthropod ectoparasites of two species of Turdus Linnaeus, 1758 (Passeriformes: Turdidae) in southern Rio Grande do Sul, Brazil.

This study was aimed at describing the community of arthropod ectoparasites associated with sympatric populations of Turdus amaurochalinus and Turdus rufiventris and analyzing the aggregation patterns of the chewing lice species, during reproductive and nonreproductive periods, of both Turdus species in three areas of the Atlantic forest in southern Rio Grande do Sul state (RS), Brazil. Altogether, we captured 36 specimens of T. amaurochalinus and 53 specimens of T. rufiventris. We identified two families of chewing lice, Menoponidae and Philopteridae, with Myrsidea and Brueelia as the most prevalent and abundant on both host birds. The lowest aggregation levels of chewing lice Myrsidea and Brueelia occurred during the reproductive period of both host species, suggesting a reproductive synchronization and a dispersion period. The most prevalent feather mite on T. amaurochalinus was Proctophyllodes weigoldi, and on T. rufiventris, Trouessartia serrana. Analges sp. and Pteronyssoides sp. were not observed on T. rufiventris. We identified three species of ticks; Ixodes auritulus was the most prevalent and abundant on the birds. Ornithoica vicina was the only hippoboscid fly collected, and only on T. amaurochalinus. The richness of ectoparasites was greater on T. amaurochalinus than on T. rufiventris. For T. amaurochalinus, the mean richness was lesser in winter compared to spring and autumn; however, we observed no variation in the mean richness of ectoparasites for T. rufiventris during the same seasons.