Magnetic Particle Spray Mass Spectrometry.

In this work, the concept of magnetic particle spray mass spectrometry (MPS-MS) is reported for the first time. Magnetic sorbent particles are used to extract the analytes from a liquid sample. The particles are magnetically attracted to the tip of a magnetic probe that is positioned at the entrance of the mass spectrometer. A solvent is dispensed on the particles, and a high voltage promotes the formation of the Taylor cone around the particles agglomerate. Analytes are desorbed by the solvent, ionized, and analyzed by mass spectrometry. MPS-MS is totally in consonance with the green chemistry principle. A minimal consumption of sample (100 µL), solvent (34 µL), and magnetic sorbent (500 µg) is needed per analysis for an excellent performance of MPS-MS in terms of sensitivity and selectivity. The determination of amitriptyline, citalopram, clomipramine, chlorpromazine, doxepin, haloperidol, nortriptyline, and venlafaxine in human plasma samples using magnetic restricted-access carbon nanotubes was carried out as a proof of principle. Limits of quantification of 10 µg L-1 and correlation coefficients higher than 0.98 were obtained for all of the analytes. Limits of detection ranged from 0.43 to 2.82 µg L-1. Precision (as relative standard deviation) and accuracies (as relative error) ranged from 3.6 to 23.6%, as well as -12.8 to 18.7%, respectively. MPS-MS opens a new line of developments in the association of sample preparation with ambient ionization. New sorbents, device configurations, and physical and chemical conditions can also be analyzed for the analysis of many other analytes in different samples.

Prenatal androgenization (2D:4D) predictions of tennis match-play success in junior players: A search for physiological explanations.

AIM: This study aims to investigate the possible association between digit ratio (2D:4D) and match-play success (MPS) in junior tennis players. In addition, we consider the possible explanatory pathways of these associations in relation to psychological, strength, power, and hormonal parameters. METHODS: We performed a cross-sectional study, with a sample comprised of 64 male junior tennis players (11-18 years old). Digit ratio was calculated from direct finger measurements. In addition, we measured the ratio of wins by number of matches played in 5 years of official competition (MPS), handgrip strength (HGS), standing long jump (SLJ), training (in weekly hours), and expertise (number of years in official competition). Salivary testosterone and cortisol levels were measured before and after physical "challenge" tests. RESULTS: The 2D:4D correlated negatively with HGS and SLJ. MPS was also negatively associated with 2D:4D, but was positively correlated to HGS, expertise, training, and self-confidence (SC). Multiple linear regression showed 2D:4D and expertise were associated with MPS (43%-54%). None of the physical, or hormonal variables tested explained the links between 2D:4D and MPS. CONCLUSION: Therefore, the specific fitness components influenced by prenatal androgenization that moderate sports success remain unknown. Future studies should explore the interaction of 2D:4D, with tennis exercises as a challenge to induce hormonal change, the effect of pubertal stage, and the influence of aerobic endurance in determining MPS.

Biofilm-producing Escherichia coli O104:H4 overcomes bile salts toxicity by expressing virulence and resistance proteins.

We investigated bile salts' ability to induce phenotypic changes in biofilm production and protein expression of pathogenic Escherichia coli strains. For this purpose, 82 pathogenic E. coli strains isolated from humans (n = 70), and animals (n = 12), were examined for their ability to form biofilms in the presence or absence of bile salts. We also identified bacterial proteins expressed in response to bile salts using sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-electrophoresis) and liquid chromatography-mass spectrometry (LC-MS/MS). Lastly, we evaluated the ability of these strains to adhere to Caco-2 epithelial cells in the presence of bile salts. Regarding biofilm formation, two strains isolated from an outbreak in Republic of Georgia in 2009 were the only ones that showed a high and moderate capacity to form biofilm in the presence of bile salts. Further, we observed that those isolates, when in the presence of bile salts, expressed different proteins identified as outer membrane proteins (i.e. OmpC), and resistance to adverse growth conditions (i.e. F0F1, HN-S, and L7/L12). We also found that these isolates exhibited high adhesion to epithelial cells in the presence of bile salts. Together, these results contribute to the phenotypic characterization of E. coli O104: H4 strains.

Can Extraintestinal Pathogenic Escherichia coli with Heat Resistance Profile Overcome Nonthermal Technologies?

Ultraviolet-C light-emitting diode (UVC-LED) and ultrasound (US) are two nonthermal technologies with the potential to destroy pathogens. However, little is known about their effectiveness in strains with a history of heat resistance. Thus, this study aimed to evaluate the phenotype and genotype of heat-resistant extraintestinal pathogenic Escherichia coli (ExPEC) with heat resistance genes after the application of US, UVC-LED, and UVC-LED+US. For this, two central composite rotatable designs were used to optimize the UVC-LED and US conditions in four ExPEC isolated from beef. From the genome of these isolates obtained in a previous study, possible genes for UVC resistance were analyzed. Results showed that US was ineffective in reducing >0.30 log colony-forming unit/mL, and that when used after UVC-LED, it showed a nonsynergic or antagonistic effect. Also, UVC-LED had the greatest effect at the maximum dose (4950 mJ/cm2 from 1.65 mW/cm2 for 50 min). However, the strains showed some recovery after that, which could be implicated in the expression of genes included in SOS system genes, some others present in the transmissible Locus of Stress Tolerance (trxBC and degP), and others (terC). Thus, ExPEC can overcome the conditions used in this study for US, UVC-LED, and UVC-LED+US, probably due to the history of resistance to other cellular damage. The result of this study will contribute to future studies that aim to find better treatment conditions for each food product.

Genomic features and heat resistance profiles of Escherichia coli isolated from Brazilian beef.

AIMS: Characterize Escherichia coli and E. coli -producing (STEC) isolates from Brazilian beef to determine heat resistance and the presence of the transmissible locus of stress tolerance (tLST). METHODS AND RESULTS: Twenty-two STEC previously isolated from beef and characterized as STEC by PCR were subjected to different heat survival challenges (60°C and 71°C). Furthermore, the three tLST-positive isolates and one tLST-negative isolate by PCR were selected for WGS analysis. Phenotypic results indicated that 3/22 (13.64%) were heat resistant, 12/22 (54.54%) were moderately resistant, and 7/22 (31.82%) were sensitive to heat treatments. WGS analyses showed that three isolates with heat resistance showed tLST with up to 80% and 42% of similarity by BLAST analysis, with the major tLST genes being responsible for the homeostasis module. However, WGS showed the absence of stx genes associated with tLST-positive isolates, albeit with virulence and resistance genes found in extraintestinal pathogenic E. coli (ExPEC). CONCLUSION: Our findings demonstrate the presence of heat-resistant E. coli as well as confirm some tLST genes in E. coli isolated from Brazilian beef.

Efficacy of Quaternary Ammonium Compounds for Control of Individual and Mixed Cultures of Escherichia coli with High- and Low-Quaternary Ammonium Compounds Resistance.

Escherichia coli is a well-characterized micro-organism in scientific literature. Similarly, quaternary ammonium compounds (QACs) are historical sanitizers in food processing. However, the use of QACs has been questioned due to bacterial resistance in some studies. Therefore, this study aimed to compare effects of single and mixed cultures of E. coli strains of different serogroups with either high (six strains) or low (five strains) resistance to QACs. Twenty-five combinations of strains with either high (H)- or low (L)-QAC resistance were analyzed (H + H vs. L + L). After exposure to QAC, combinations with statistical differences (p < 0.05) compared with individuals were selected and an inactivation model determined using GInaFit®. Only one combination of two strains (C23 and C20) with low-QAC resistance (mixture T18) had greater resistance (p < 0.05) than the individual isolates. The combination T18 and individual strain C23 presented a Weibull model, whereas the other isolated strain (C20) presented a biphasic inactivation model with a shoulder. Whole genome sequencing determined that unlike C20, C23 carried yehW, which may have led to Weibull inactivation. Possibly, very rapid interaction of C20 with the QAC favored increased survival of C23 and overall persistence of the T18 mixture. Consequently, our results indicate that individual E. coli with low-QAC resistance can synergistically interfere with QAC inactivation.

Online restricted access molecularly imprinted solid-phase extraction coupled with liquid chromatography-mass spectrometry for the selective determination of serum bile acids.

A rapid and sensitive online restricted access molecularly imprinted solid-phase extraction method coupled to a liquid chromatography-mass spectrometry (LC-MS) system for simultaneous determination of serum bile acids as well as their taurine and glycine conjugates was developed. Reversible liver damage of workers exposed to volatile organic solvents can be investigated based on the level of the analyzed molecules. A restricted access molecularly imprinted polymer coated with bovine serum albumin (RAMIP-BSA) was synthesized and used as the extraction phase. The column switching liquid chromatography system was able to exclude about 100% of the macromolecules and extract/separate nine bile acids from blood human serum samples, in a total time of 40 minutes. The developed method was validated based on the Food and Drug Administration (FDA) guidelines, being linear for all the analytes in their respective analytical ranges (coefficients of determination higher than 0.99), with limits of detection (LOD) and quantification (LOQ) ranging from 2.0 to 5.7 µg L-1 and from 10.0 to 25.0 µg L-1, respectively. Suitable results for precision (relative standard deviation ranged from 3.2 to 14.5% and 0.7 to 14.8%, respectively for intra and inter-assay) and accuracy (relative error ranged from -14.8 to 14.2%; -13.8 to 14.3%) were obtained. The validated analytical method was applied to determine bile acids in serum samples of five workers occupationally exposed to volatile organic solvent, demonstrating its applicability in the assessment of these toxicants.

Antimicrobial Resistance of Listeria monocytogenes from Animal Foods to First- and Second-Line Drugs in the Treatment of Listeriosis from 2008 to 2021: A Systematic Review and Meta-Analysis.

First-line drugs for the treatment of listeriosis are the same around the world, but particular conditions might reduce their efficacy, including antimicrobial resistance. Therefore, this study aimed to verify, based on a systematic review and meta-analysis, whether the prevalence of antimicrobial resistance in Listeria monocytogenes from animal foods is higher for first- or second-line antimicrobials. From the total of 302 identified studies, 16 met all the eligibility criteria from 2008 to 2021 and were included in this meta-analysis. They comprised a dataset of 1152 L. monocytogenes isolates, obtained from different animal food products, food processing environment, and live animals. The included studies were developed in South America (n = 5), Europe (n = 4), Asia (n = 3), Africa (n = 2), and North America (n = 2), testing a total of 35 different antimicrobials, 11 of them classified as first-line drugs. Complete lack of antimicrobial resistance across the studies (all L. monocytogenes isolates tested as susceptible) was only observed for linezolid, while widespread antimicrobial resistance (all L. monocytogenes isolates tested resistant) was described for amoxicillin, benzylpenicillin, cefoxitin, fusidic acid, imipenem, sulfamethoxazole, and vancomycin. Overall, the meta-analysis results indicated no evidence that antimicrobial resistance in L. monocytogenes isolated from animal-based food is higher for first-line antimicrobials compared to second-line compounds (p=0.37). A greater volume of publication, together with better characterization of the isolates, is still needed for a more precise estimate of the real prevalence of antimicrobial resistance in L. monocytogenes.

Shiga toxin-producing Escherichia coli isolated from pasteurized dairy products from Bahia, Brazil.

The presence of pathogenic Shiga toxin-producing Escherichia coli (STEC) in dairy products represents a public health concern because of its ability to produce the toxins Stx1 and Stx2, which cause intestinal diseases. Monitoring the stages of milk production and checking dairy products for contamination are crucial steps to ensure dairy safety. This study aimed to report the occurrence of thermotolerant coliforms, E. coli, and STEC strains in pasteurized dairy products and to evaluate the antibiotic resistance profiles, serotypes, and characterizations of the STEC isolates by pulsed-field gel electrophoresis. We obtained a total of 138 pasteurized dairy products from 15 processing plants in Bahia, Brazil, to examine coliforms, E. coli, and STEC strains. We found that 43% of samples (59/138) contained thermotolerant coliforms, and 30% (42/138) did not comply with Brazilian regulations. Overall, 6% (9/138) were positive for E. coli and 4% (5/138) were positive for STEC. We recovered 9 STEC isolates from pasteurized cream (2/9), Minas Padrão cheese (2/9), Minas Frescal cheese (4/9), and ricotta (1/9). All isolates were stx2-positive, and 2 were eae-positive. All isolates were negative for the "big 6" STEC serogroups, belonging instead to serotypes ONT:HNT, ONT:H12, O148:H-, OR:H40, OR:HNT, and O148:HNT. Pulsed-field gel electrophoresis revealed 100% genetic similarity among 3 isolates from 2 different samples produced in the same production facility, which may suggest cross-contamination. As well, we found isolates that were 98% similar but in samples produced in different production facilities, suggesting a mutual source of contamination or a circulating strain. Two STEC strains exhibited resistance to streptomycin. Although the isolates presented a low resistance profile and no strain belonged to the "big 6" pathogenic group, the circulation of stx2-positive STEC strains in ready-to-eat products highlights the importance of epidemiological surveillance inside the Brazilian dairy chain.

Salmonella in the processing line of farmed Tambatinga (Colossoma macropomum x Piaractus brachypomus) in Mato Grosso, Brazil: serotypes of occurrence and antimicrobial profile.

The objective of this study was to evaluate the dispersion dynamics and antimicrobial resistance profiles of Salmonella in the processing of Tambatinga (Colossoma macropomum x Piaractus brachypomus). Thirty fish were monitored during four processing stages (reception, first wash, evisceration, and prepackage area) in a fish slaughterhouse. One hundred and twenty fish surface samples were collected and tested through bacteriological analysis, PCR, serotyping, and antimicrobial resistance profile (disk-diffusion). Of these samples, 7.5% (9/120) were positive for Salmonella, with 0.83% being observed in the pre-packaging phase, indicating a low occurrence at this stage. All the analyzed stages were positive for Salmonella, with the prevalent serovars being Ndolo, Mbandaka, Typhimurium, Rough, and O:16. All strains were sensitive to various antimicrobials. Improvements in microbiological control during all processing stages should be implemented to ensure a Salmonella-free product.

The impact of exercise training on calf pump function, muscle strength, ankle range of motion, and health-related quality of life in patients with chronic venous insufficiency at different stages of severity: a systematic review.

Exercise training (ET) is an important tool in the management of patients with chronic venous insufficiency (CVI). The objective of this article was to discuss the effects of ET on the calf pump, functional parameters, and quality of life of patients with mild and advanced CVI. A systematic review was conducted and eleven studies were included. In patients with mild CVI, ET was effective for improving venous reflux, muscle strength, ankle range of motion, and quality of life. In advanced CVI patients, ET increased ejection fraction, reduced residual volume fraction, and improved muscle strength and ankle range of motion, but did not change venous reflux indices or quality of life. It is concluded that ET is effective for improving calf pump function, muscle strength, and ankle range of motion in CVI. In patients with mild CVI, additional benefits were observed in quality of life.

O treinamento físico é uma importante ferramenta no tratamento de pacientes com insuficiência venosa crônica. O objetivo foi discutir os efeitos do tratamento físico na bomba da panturrilha, os parâmetros funcionais e a qualidade de vida de pacientes com insuficiência venosa crônica leve e avançada. Uma revisão sistemática foi realizada, e 11 estudos foram incluídos. Na insuficiência venosa crônica leve, o treinamento físico foi eficaz na melhora do refluxo venoso, da força muscular, da amplitude de movimento do tornozelo e da qualidade de vida. Na insuficiência venosa crônica avançada, o treinamento físico aumentou a fração de ejeção, reduziu a fração de volume residual e melhorou a força muscular e amplitude de movimento do tornozelo, sem alterações nos índices de refluxo venoso e na qualidade de vida. Conclui-se que o treinamento físico é eficaz na melhoria da bomba da panturrilha, da força muscular e da amplitude de movimento do tornozelo na insuficiência venosa crônica. Em pacientes com insuficiência venosa crônica leve, foram encontrados benefícios adicionais na qualidade de vida.

Restricted access carbon nanotube for microextraction by packed sorbent to determine antipsychotics in plasma samples by high-performance liquid chromatography-tandem mass spectrometry.

This manuscript describes the development of the restricted access carbon nanotube (RACNT) as a selective stationary phase for microextraction by packed sorbent (MEPS) to determine antipsychotics (chlorpromazine, clozapine, olanzapine, and quetiapine) in untreated plasma samples from schizophrenic patients by ultra-high liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). The synthesis was achieved by chemically covering commercial multi-walled carbon nanotubes with bovine serum albumin (BSA) to subsequently pack the material in a polyethylene conical tube (1000 µL). The RACNTs' sorbents were able to exclude about 97% of the plasma proteins, maintaining the same performance for about 100 assays. The MEPS variables (sample pH, draw-eject cycles, desorption and phase cleanup) were evaluated to improve sensibility and selectivity. The MEPS/UHPLC-MS/MS method was linear at concentrations ranging from the lower limit of quantification (10.0 ng mL-1) to the upper limit of quantification (200-700 ng mL-1) with coefficients of determinations higher than 0.99. The precision assays presented relative standard deviation (RSD) values lower than 13%, and the accuracy assays presented relative error (RE) values that ranged from - 8.01 to 11.53%. Neither significant matrix effects nor carryover was observed. The developed method was successfully applied to determine antipsychotics drugs for therapeutic drug monitoring of schizophrenic patients.

Worldwide Epidemiology of Salmonella Serovars in Animal-Based Foods: a Meta-analysis.

Salmonella spp. are among the most important foodborne pathogens and the third leading cause of human death among diarrheal diseases worldwide. Animals are the primary source of this pathogen, and animal-based foods are the main transmission route to humans. Thus, understanding the global epidemiology of Salmonella serovars is key to controlling and monitoring this bacterium. In this context, this study aimed to evaluate the prevalence and diversity of Salmonella enterica serovars in animal-based foods (beef, pork, poultry, and seafood) throughout the five continents (Africa, the Americas [North and Latin America], Asia, Europe, and Oceania). The meta-analysis consisted of a chemometric assessment (hierarchical cluster analysis and principal component analysis) to identify the main epidemiological findings, including the prevalence and diversity of the Salmonella serovars in each matrix. Regarding the serovar distribution, S Typhimurium presented a cosmopolitan distribution, reported in all four assessed matrices and continents; poultry continues to play a central role in the dissemination of the Enteritidis serovar to humans, and Anatum and Weltevreden were the most frequently found in beef and seafood, respectively. Additionally, we recommended careful monitoring of certain serovars, such as Derby, Agona, Infantis, and Kentucky. Finally, given the scientific data regarding the most frequently reported serovars and which matrices constitute the main vehicles for the transmission of this pathogen, control programs may be improved, and specific interventions may be implemented in an attempt to reduce the risk of this pathogen reaching humans.IMPORTANCE Salmonellosis is caused by Salmonella spp. and is the third leading cause of death among food-transmitted diseases. This pathogen is commonly disseminated in domestic and wild animals, and the infection's symptoms are characterized by acute fever, nausea, abdominal pain, and diarrhea. The animals are the primary source of salmonellae, and animal-based foods are the main transmission route to humans. Therefore, data collected from these sources could contribute to future global interventions for effective control and surveillance of Salmonella along the food chain. In light of this, the importance of our research is in identifying the prevalence of Salmonella serovars in four animal-based food matrices (pork, poultry, beef, and seafood) and to evaluate the importance that each matrix has as the primary source of this pathogen to humans.

A new restricted access molecularly imprinted fiber for direct solid phase microextraction of benzodiazepines from plasma samples.

Restricted access molecularly imprinted polymers (RAMIPs) are hybrid materials that present selective binding sites for a template (or similar molecules), and an external hydrophilic layer that avoids the binding of proteins to the material, making them appropriate for the sample preparation of protein fluids. RAMIPs have been used successfully in online and offline solid phase extractions, but there is no application as a fiber in solid phase microextraction (SPME), to the best of our knowledge. In this paper, molecularly imprinted fibers were synthesized inside glass capillary tubes (0.53 mm i.d.), using diazepam and methacrylic acid as template and functional monomer, respectively. The MIP fibers were coated with a cross-linked bovine serum albumin (BSA) layer, resulting in RAMIP fibers that were used in the SPME of benzodiazepines directly from biological fluids. The BSA layer acts as a protective barrier that avoids the binding of proteins from the sample by an electrostatic repulsion mechanism. The protein exclusion capacity of the RAMIP fiber was about 98%, which is selective to benzodiazepines in comparison with other drugs (citalopram and fluoxetine). The SPME was optimized and the extraction conditions were set as follows: 1000 µL of the sample diluted with water (1 : 0.5, v : v), no pH adjustment, an extraction time of 20 min at 500 rpm, and elution with 200 µL of acetonitrile for 5 min at 500 rpm. The fibers were used in the SPME of benzodiazepines directly from plasma samples, followed by HPLC-DAD analyses. The method was linear for bromazepam (50-750 µg L-1), clonazepam (15-250 µg L-1), alprazolam (15-350 µg L-1), nordiazepam (100-2100 µg L-1) and diazepam (100-2600 µg L-1), with correlation coefficients higher than 0.97. Relative standard deviations (precision) and relative errors (accuracy) ranged from 0.5 to 20.0%, and -15.6 to 21.6%, respectively.

Biological sample preparation by using restricted-access nanoparticles prepared from bovine serum albumin: application to liquid chromatographic determination of ß-blockers.

Restricted-access nanoparticles (RANPs) were prepared from bovine serum albumin by coacervation. They have an average sized of 311 nm. They were characterized and used to capture the ß-blockers atenolol, metoprolol and propranolol from untreated biological samples. It is shown that both high protein affinity drugs (propranolol) and low protein affinity drugs (atenolol) could be rapidly extracted from plasma. This is revealed by kinetic and isothermal adsorption studies. On the other hand, almost all proteins from the sample were excluded. This demonstrates the efficiency of RANPs as restricted-access material. Sample preparation was carried out by solid phase microextraction using a probe obtained by the fixation of the RANPs at the end of a glass capillary. Atenolol (in concentrations from 100 to 1200 µg L-1), metoprolol (from 80 to 1000 µg L-1) and propranolol (from 15 to 200 µg L-1) were extracted from spiked plasma samples and analyzed by LC MS/MS without using a separation column. Correlation coefficients >0.99, good precision, accuracy, robustness, and lack of memory effects were observed for all of the analytes. The detection limits (at an S/N of 3) are 25.6, 14.6, and 3.8 µg L-1 for atenolol, metoprolol and propranolol, respectively. Ten samples can be simultaneously extracted within ∼15 min. Plasma samples of patients undergoing medical treatment were successfully analyzed with the method. Graphical abstract Schematic representation of a bovine serum albumin-based restricted access nanoparticle that exclude proteins from a human plasma sample but capture the small analytes.

Occurrence and antimicrobial resistance of E. coli non-O157 isolated from beef in Mato Grosso, Brazil.

Shiga toxin-producing Escherichia coli (STEC) is an important public health concern pathogen, as it produces two toxins, Stx1 and Stx2, with cytotoxic capacity. In addition, STEC strains are frequently involved in food outbreaks worldwide, leading to public health challenges and economic losses. In this context, the occurrence and antimicrobial resistance profile of the STEC isolated from fresh beef produced in Mato Grosso, Brazil, were estimated. One hundred seven retail beef under vacuum-packaged produced by 13 different slaughterhouses were submitted to microbiological, molecular, and antimicrobial resistance analyses. STEC occurrence in beef was of 4.67%, and five strains presented the stx2 gene. The O111 serogroup, reported in several outbreak cases worldwide, was detected, and other serotypes (O8:H20, O22:H16, and O141:H49) were also isolated. All isolated strains displayed sensitivity to 12 antibiotics, except for two strains, which where streptomycin-resistant. The presence of STEC in retail beef samples indicates public health risks with significant economic impact throughout the retail beef chain.

Acetic Acid Increased the Inactivation of Multi-drug Resistant Non-typhoidal Salmonella by Large-Scaffold Antibiotic.

Salmonella is a gram-negative bacterium with intrinsic resistance to large-scaffold antibiotics due to the presence of an outer membrane. Based on the mode of action of the organic acids in outer membrane disintegration, and consequently, an enhancement in cell permeability, a combination of acetic acid and a large-scaffold antibiotic is it evaluated. Therefore, the aim of this study is to assess the combination of different levels of acetic acid with vancomycin, in order to determine whether or not the organic acid may overcome the cell wall and the intrinsic resistance in multi-drug resistant Salmonella. Screening of five wild-type Salmonella strains and one clinical strain was performed to select the strain more resistance to acid inhibition. Acetic acid was tested at 2.0, 1.75, 1.50, and 1.25% levels, separated or combined with 8 µg/mL vancomycin dose. An aliquot was collected after exposure and inoculated into the brain and heart infusion agar. The plates were counted and the data analyzed by ANOVA and a posthoc Tukey test (p < 0.05). The results indicate that 1.25 and 1.50% levels did not affect the vancomycin inactivation of multi-drug resistant Salmonella. However, at levels of 1.75 and 2.0%, an increase in microbial reduction is observed. Also, 2% level acetic acid and vancomycin had a threefold increase compared to vancomycin alone. Therefore, the use of acetic acid as prior treatment for Salmonella increased the inactivation rate of vancomycin. The combination of organic acid and antibiotics is a potential tool to overcome cases of antimicrobial resistance.

Escherichia coli O26 and O113:H21 on Carcasses and Beef from a Slaughterhouse Located in Mato Grosso, Brazil.

Shiga toxin-producing Escherichia coli (STEC) is a group of emerging pathogens that can cause human diseases, including hemolytic uremic syndrome (HUS) and hemorrhagic colitis (HC). Monitoring slaughtering stages and checking contamination points are crucial for the production of safe food. In this context, the aim of this study was to verify contamination by STEC strains, to determine the contamination points and evaluate the resistance profile to 12 antimicrobials used in both veterinary and human medicine. A total of 80 samples were obtained from eight collection points (pen floor, rectum, hide, carcass swabs and esophagus, diaphragm, masseter, and retail beef tissue samples). The isolates were collected by dilution plating on MacConkey agar with sorbitol, cefixime, and tellurite and analyzed by multiplex polymerase chain reaction for virulence genes. Serotyping of non-O157 was performed, and testing for 12 antibiotics by disk diffusion was carried out. A total of 18 STEC strains were isolated, presenting different virulence profiles. Contamination by STEC was observed in the rectum (5/18), carcass surface (5/18), hide (3/18), diaphragm (2/18), retail beef (2/18), and masseter muscle (1/18). Pen floor swabs and esophagus tissues showed no STEC contamination. Moreover, three strains were identified as O26 and three as O113:H21 strains, which have been linked to HUS and HC outbreak cases in Brazil. All STEC isolates were susceptible to all evaluated antimicrobials, except streptomycin. The presence of STEC strains is a direct risk to the consumer, especially when isolated from retail beef, and contamination can occur during different slaughter stages. However, antimicrobial resistance profiles did not identify multidrug-resistant strains, limiting potential antimicrobial resistance transmission to other pathogens.

Molecularly imprinted probe for solid-phase extraction of hippuric and 4-methylhippuric acids directly from human urine samples followed by MEKC analysis.

Hippuric acid (HA) and 4-methylhippuric acid (4-MHA) are metabolites as well as biological indicators for toluene and xylenes, respectively, and their determination in urine samples is very important, in order to monitor the occupational exposition to these solvents, ensuring a safe working environment. Thus, this paper describes the synthesis and characterization of a probe impregnated with molecularly imprinted polymers (MIPs) for the solid-phase extraction of HA and 4-MHA directly from untreated urine samples followed by micellar electrokinetic chromatography (MEKC) analyses. The MIP probe selectivity was compared to the non-imprinted polymer probe. The MEKC separations were carried out in 50 mmol/L sodium tetraborate pH 10.0/0.5 mmol/L cetyltrimethylammonium bromide aqueous solution, with a constant voltage of -15 kV. The system variables were optimized to provide ideal conditions for the extraction and desorption of the analytes, as well as for the MEKC analyses. The method was linear from 0.5 to 5.0 g/L for both analytes, with correlation coefficients > 0.994. Precisions and accuracies, expressed as relative standard deviation and relative error, were < 20.0 and within -15.4 to 16.6%, respectively, in accordance with the United States Food and Drug Administration recommendation. The MIP probe has proven to be simple, cheap, resistant, and synthetically reproducible, being successfully used to analyze both HA and 4-MHA from real samples.

Differential expression of extracellular matrix genes in glenohumeral capsule of shoulder instability patients.

Anterior shoulder instability is a common orthopedic problem. After a traumatic shoulder dislocation, patients present a plastic deformation of the capsule. The shoulder instability biology remains poorly understood. We evaluated the expression of genes that encode the cartilage oligomeric matrix protein (COMP), fibronectin 1 (FN1), tenascin C (TNC) and tenascin XB (TNXB) in the glenohumeral capsule of anterior shoulder instability patients and controls. Moreover, we investigated the associations between gene expression and clinical parameters. The gene expression was evaluated by quantitative reverse transcription-polymerase chain reaction in the antero-inferior (macroscopically injured region), antero-superior and posterior regions of the capsule of 29 patients with shoulder instability and 8 controls. COMP expression was reduced and FN1 and TNC expression was increased in the antero-inferior capsule region of cases compared to controls (p < 0.05). TNC expression was increased in the posterior capsule portion of shoulder instability patients (p = 0.022). COMP expression was reduced in the antero-inferior region compared to the posterior region of shoulder instability patients (p = 0.007). In the antero-inferior region, FN1 expression was increased in the capsule of patients with more than one year of symptoms (p = 0.003) and with recurrent dislocations (p = 0.004) compared with controls. FN1 and TNXB expression was correlated with the duration of symptoms in the posterior region (p < 0.05). Thus, COMP, FN1, TNC and TNXB expression was altered across the capsule of shoulder instability patients. Dislocation episodes modify FN1, TNC and TNXB expression in the injured tissue. COMP altered expression may be associated with capsule integrity after shoulder dislocation, particularly in the macroscopically injured portion.