Development of Phenothiazine Hybrids with Potential Medicinal Interest: A Review.

The molecular hybridization approach has been used to develop compounds with improved efficacy by combining two or more pharmacophores of bioactive scaffolds. In this context, hybridization of various relevant pharmacophores with phenothiazine derivatives has resulted in pertinent compounds with diverse biological activities, interacting with specific or multiple targets. In fact, the development of new drugs or drug candidates based on phenothiazine system has been a promising approach due to the diverse activities associated with this tricyclic system, traditionally present in compounds with antipsychotic, antihistaminic and antimuscarinic effects. Actually, the pharmacological actions of phenothiazine hybrids include promising antibacterial, antifungal, anticancer, anti-inflammatory, antimalarial, analgesic and multi-drug resistance reversal properties. The present review summarizes the progress in the development of phenothiazine hybrids and their biological activity.

Trends in microbial control techniques for poultry products.

Fresh poultry meat and poultry products are highly perishable foods and high potential sources of human infection due to the presence of several foodborne pathogens. Focusing on the microbial control of poultry products, the food industry generally implements numerous preventive measures based on the Hazard Analysis and Critical Control Points (HACCP) food safety management system certification together with technological steps, such as refrigeration coupled to modified atmosphere packaging that are able to control identified potential microbial hazards during food processing. However, in recent years, to meet the demand of consumers for minimally processed, high-quality, and additive-free foods, technologies are emerging associated with nonthermal microbial inactivation, such as high hydrostatic pressure, irradiation, and natural alternatives, such as biopreservation or the incorporation of natural preservatives in packaging materials. These technologies are discussed throughout this article, emphasizing their pros and cons regarding the control of poultry microbiota and their effects on poultry sensory properties. The discussion for each of the preservation techniques mentioned will be provided with as much detail as the data and studies provided in the literature for poultry meat and products allow. These new approaches, on their own, have proved to be effective against a wide range of microorganisms in poultry meat. However, since some of these emergent technologies still do not have full consumer's acceptability and, taking into consideration the hurdle technology concept for poultry processing, it is suggested that they will be used as combined treatments or, more frequently, in combination with modified atmosphere packaging.

Antimicrobial activity of coriander oil and its effectiveness as food preservative.

ABTRACT Foodborne illness represents a major economic burden worldwide and a serious public health threat, with around 48 million people affected and 3,000 death each year only in the USA. One of the possible strategies to reduce foodborne infections is the development of effective preservation strategies capable of eradicating microbial contamination of foods. Over the last years, new challenges for the food industry have arisen such as the increase of antimicrobial resistance of foodborne pathogens to common preservatives and consumers demand for naturally based products. In order to overcome this, new approaches using natural or bio-based products as food preservatives need to be investigated. Coriander (Coriandrum sativum L.) is a well-known herb widely used as spice, or in folk medicine, and in the pharmacy and food industries. Coriander seed oil is the world's second most relevant essential oil, exhibiting antimicrobial activity against Gram-positive and Gram-negative bacteria, some yeasts, dermatophytes and filamentous fungi. This review highlights coriander oil antimicrobial activity and possible mechanisms of action in microbial cells and discusses the ability of coriander oil usage as a food preservative, pointing out possible paths for the successful evolution for these strategies towards a successful development of a food preservation strategy using coriander oil.

Development of carboxymethyl xylan films with functional properties.

The aim of this work was focused on the development of carboxymethyl xylan (CMX) formulations with functional properties to produce edible films. Beechwood Xylan was firstly derivatized into carboxymethyl xylan and thereafter was blended with Agar (Ag), Ammonium zirconium carbonate (AZC) and linoleic acid (La) to produce CMX:Ag, CMX:AZC, CMX:Ag:La films. Mechanical, barrier, optical and thermal properties of the produced films and their antimicrobial activity against food pathogenic bacteria were evaluated. The obtained films were transparent and yellowish. Agar and AZC improved the tensile strength at break of the control CMX film from 4.79 to 27.67 and 20.95 MPa respectively, and the CMX:AZC film exhibited the greatest elastic modulus. Barrier properties of the films decreased when any of the components was incorporated into the CMX and all blended films were thermally more stable than control. The CMX:Ag:La film revealed a good antimicrobial activity against B. cereus and S. aureus.

Insights in the pathogenesis and resistance of Arcobacter: A review.

Arcobacter genus currently comprises 18 recognized species, among which Arcobacter butzleri, Arcobacter cryaerophilus and Arcobacter skirrowii have been associated with human and animal disease. Although these organisms, with special emphasis A. butzleri, are emerging as clinical pathogens, several aspects of their epidemiology and virulence are only starting to be clarified. In vitro human and animal cell culture assays have been used to show that several Arcobacter species can adhere to and invade eukaryotic cells, induce an immune response and produce toxins that damage host cells. In addition, data from genome sequencing highlighted several potential markers that may be helpful candidates for the study and understanding of these mechanisms; however, more work is necessary to clarify the molecular mechanisms involved in Arcobacter virulence. Arcobacter can be considered a relatively robust organism showing to be able to survive in adverse conditions, as the ones imposed by food processing and storage. Moreover, these bacteria have shown increased antibiotic resistance, along with high multidrug resistance. In this review, we seek to update the state-of-the-art concerning Arcobacter distribution, its interaction with the host, the trends of antibiotic resistance, its ability to survive, and finally the use of natural antimicrobials for control of Arcobacter.

Study of the major essential oil compounds of Coriandrum sativum against Acinetobacter baumannii and the effect of linalool on adhesion, biofilms and quorum sensing.

Acinetobacter baumannii is a pathogen that has the ability to adhere to surfaces in the hospital environment and to form biofilms which are increasingly resistant to antimicrobial agents. The aim of this work was to study the antimicrobial activity of the major oil compounds of Coriandrum sativum against A. baumannii. The effect of linalool on planktonic cells and biofilms of A. baumannii on different surfaces, as well as its effect on adhesion and quorum sensing was evaluated. From all the compounds evaluated, linalool was the compound with the best antibacterial activity, with minimum inhibitory concentration values between 2 and 8 µl ml(-1). Linalool also inhibited biofilm formation and dispersed established biofilms of A. baumannii, changed the adhesion of A. baumannii to surfaces and interfered with the quorum- sensing system. Thus, linalool could be a promising antimicrobial agent for controlling planktonic cells and biofilms of A. baumannii.

Genotypic and phenotypic features of Arcobacter butzleri pathogenicity.

Even though Arcobacter butzleri has been implicated in some human disease as diarrhoea and bacteraemia, much of its pathogenesis and virulence factors remain unclear. In this work we have compared pathogenic and genotypic properties of six A. butzleri isolates from human and non-human sources. The tested isolates showed to be susceptible to tetracyclines and aminoglycosides, however non-human isolates were all resistant to quinolones. The ability to form biofilms was variable among the tested strains, and all of them showed a weak haemolytic activity. The presence of nine putative virulence genes was determined, with cadF, ciaB, cj1349, mviN, pldA, tlyA being detected in all strains, while irgA (3/6), hecA (5/6), hecB (4/6) were detected only in some strains. High levels of adhesion were observed for A. butzleri on Caco-2 cells, with pre-existing inflammation showing no significant effect on the adherence ability; yet variable levels of invasion were observed. A. butzleri isolates were able to survive intracellularly in Caco-2 cells and to induce a significant up-regulation of interleukin-8 secretion and structural cell rearrangements. These data brings new insights on A. butzleri virulence and highlights its pathogenic potential.

Evaluation of the prebiotic potential of arabinoxylans from brewer's spent grain.

Arabinoxylans (AX) consumption has been related to the treatment and prevention of cardiovascular diseases, type II diabetes, colorectal cancer and obesity. The beneficial health effects are conferred through gut microbiota modulation, and therefore, they have been proposed as potential slowly fermentable prebiotic candidates. As the mechanisms are not yet well understood, the prebiotic potential of AX from brewer's spent grain (BSG) has been investigated. Two types of AX from BSG (AX1 and AX2) of different length and branching averages were fermented with human faecal inocula and compared to fermented cultures containing a commercial prebiotic (fructooligosaccharide (FOS)) and cultures with no added carbohydrate (control). Results demonstrated that the AX were extensively metabolised after 48 h of fermentation. The pH decreased along fermentation and the lowest value was achieved in AX1 cultures. The production of short chain fatty acids (SCFA) was higher in AX cultures than in cultures containing FOS and controls, with AX1 presenting the highest concentrations. The stimulatory effect of beneficial bacteria was higher in AX cultures, and AX2 presented the highest positive effect. Prebiotic potential of AX from BSG was confirmed by the production of SCFA and the modulation of gut microbiota, especially by the high increase in bifidobacteria populations.

Linalool Reduces Virulence and Tolerance to Adverse Conditions of Listeria monocytogenes.

Listeria monocytogenes, a foodborne pathogen causing listeriosis, poses substantial societal, economic, and public health challenges due to its resistance, persistence, and biofilm formation in the food industry. Exploring subinhibitory concentrations of compounds to target virulence inhibition and increase susceptibility to adverse conditions presents a promising strategy to mitigate its impact of L. monocytogenes and unveils new potential applications. Thus, this study aims to explore the effect of linalool on virulence factors of L. monocytogenes and potential use in the reduction in its tolerance to stressful conditions. This action was analysed considering the use of two sub-inhibitory concentrations of linalool, 0.312 and 0.625 mg/mL. We found that even with the lowest tested concentrations, a 65% inhibition of violacein production by Chromobacterium violaceum, 55% inhibition in biofilm formation by L. monocytogenes and 62% reduction on haemolysis caused by this bacterium were observed. In addition to its impact on virulence factors, linalool diminished the tolerance to osmotic stress (up to 4.3 log reduction after 24 h with 12% NaCl), as well as to high (up to 3.8 log reduction after 15 min at 55 °C) and low temperatures (up to 4.6 log reduction after 84 days with 12% NaCl at 4 °C). Thus, this study paves the way to further investigation into the potential utilization of linalool to mitigate the threat posed by L. monocytogenes in the field of food safety and public health.

Plasmid DNA fermentation strategies: influence on plasmid stability and cell physiology.

In order to provide sufficient pharmaceutical-grade plasmid DNA material, it is essential to gain a comprehensive knowledge of the bioprocesses involved; so, the development of protocols and techniques that allow a fast monitoring of process performance is a valuable tool for bioprocess design. Regarding plasmid DNA production, the metabolic stress of the host strain as well as plasmid stability have been identified as two of the key parameters that greatly influence plasmid DNA yields. The present work describes the impact of batch and fed-batch fermentations using different C/N ratios and different feeding profiles on cell physiology and plasmid stability, investigating the potential of these two monitoring techniques as valuable tools for bioprocess development and design. The results obtained in batch fermentations showed that plasmid copy number values suffered a pronounced increase at the end of almost all fermentation conditions tested. Regarding fed-batch fermentations, the strategies with exponential feeding profiles, in contrast with those with constant feeding, showed higher biomass and plasmid yields, the maximum values obtained for these two parameters being 95.64 OD(600) and 344.3 mg plasmid DNA (pDNA)/L, respectively, when using an exponential feed rate of 0.2 h(-1). Despite the results obtained, cell physiology and plasmid stability monitoring revealed that, although higher pDNA overall yields were obtained, this fermentation exhibited lower plasmid stability and percentage of viable cells. In conclusion, this study allowed clarifying the bioprocess performance based on cell physiology and plasmid stability assessment, allowing improvement of the overall process and not only plasmid DNA yield and cell growth.

Effect of soft tissue noise on trabecular bone score in postmenopausal women with diabetes: A cross sectional study.

BACKGROUND: Type 2 diabetes (T2D) is associated with increased fracture risk, despite similar or greater BMD compared to nondiabetics. TBS predicts fracture risk in T2D and nondiabetics. However, increased abdominal thickness, a common feature in T2D, may reduce TBS values. AIM: To study the relationship among glycemic status, BMD and TBS, considering abdominal soft tissue thickness (STT) interference. METHODS: Cross-sectional analysis of 493 women ≥65 years, with simultaneous DXA scans and HbA1c measures. STT and TBS (iNsight Software, v3.0) were derived from lumbar spine (LS) scans. Subjects were divided according to HbA1c levels: 1 (≥6.5%; n = 116), 2 (5.7-6.4%; n = 217) and 3 (≤5.6%; n = 160). Group 1 was further divided based on HbA1c and/or disease duration: 1a (HbA1c ≥ 7.5%; n = 42), 1b (HbA1c ≥ 6.5% and disease duration ≥5 years; n = 63) and 1c (HbA1c ≥ 7.5% and disease duration ≥5 years; n = 30). FINDINGS: For the entire cohort, mean age, TBS, BMI and STT were 71.8 ± 6.0 years, 1.299 ± 0.101, 26.9 ± 4.1 kg/m2, and 21.4 ± 2.9 cm, respectively. LS-BMD was similar among groups. BMD in hip sites and STT were higher in group 1. TBS was lower in patients with higher HbA1c (P = 0.020), with a mean TBS in groups 1, 2, and 3 of 1.280, 1.299 and 1.314, respectively. This difference remained after adjusting for age, LS-BMD and BMI (P = 0.010). After replacing BMI with STT, TBS differences were no longer significant (P = 0.270). The same was observed when subgroups 1a and 1b were compared to group 3. However, for subgroup 1c, TBS remained lower compared to group 3, even after adjusting for age, LS-BMD and STT, with a borderline P-value (1.275 vs. 1.308; P = 0.047). CONCLUSION: Higher HbA1c levels were associated with greater BMD in hip sites, higher abdominal STT and lower TBS values. However, after including the STT in the adjustment, TBS differences among groups disappeared, except in women with higher HbA1c levels and longer disease duration.

Influence of growth conditions on plasmid DNA production.

The obtention of high yields of purified plasmid DNA is viewed as an essential issue to be considered towards efficient production of DNA vaccines and therapeutic plasmids. In this work, Escherichia coli DH5alpha bearing the pVAX1-LacZ plasmid was grown in a developed semi-defined medium at different temperatures and tryptone concentrations. Analysis of pDNA yields and E. coli morphology revealed that at higher temperatures (37 and 40 degrees C), higher specific yields and E. coli filamentation were obtained. However, the best results were achieved when a lower tryptone concentration was used. This approach was shown to be a powerful tool to promote plasmid amplification, keeping the desirable plasmid structure, and favoring the attainment of quality. Our results suggest that by using tryptone alone as an amino acid source, pDNA amplification was improved and a specific yield of 20.43 mg pDNA/g dcw was achieved, proving that this strategy can improve pDNA yield even at a small scale.

Encapsulation of cinnamon oil in cyclodextrin nanosponges and their potential use for antimicrobial food packaging.

The main goal of this work is the encapsulation of cinnamon essential oil in cyclodextrin nanosponges and the assessment of their antimicrobial activity against foodborne pathogens. After nanosponge synthesis, a headspace-solid phase microextraction coupled to gas chromatography-mass spectrometry (HS-SPME-GC-MS) method was validated to quantify essential oil major compounds. Results showed that essential oil was successfully encapsulated in cyclodextrin nanosponges with α-NS and ß-NS being able to encapsulate higher essential oil amounts. Cinnamon essential oil, alone and encapsulated in nanosponges, proved to have antimicrobial activity against foodborne bacteria. Time-kill assays proved that the essential oil, alone or encapsulated, had a bacteriostatic effect against all bacteria tested, with the exception of Y. enterocolitica where a bactericidal action was observed. Furthermore, the controlled release achieved by its encapsulation, allowed cinnamon essential oil to be effective at a much lower concentration in culture medium than when solely dissolved in culture medium. Thus, the results described herein encourage the use of cyclodextrin nanosponges as encapsulating agents for active food packaging applications.

A meta-analytic perspective on Arcobacter spp. antibiotic resistance.

OBJECTIVES: Over the years, an increased prevalence of resistant strains of Arcobacter has been observed, which may be due to Arcobacter exposure to antibiotics used both in animal production and human medicine. A systematic review was performed with the objective of summarising the results of the rates of antimicrobial resistance of Arcobacter isolates. METHODS: The systematic review was performed according to PRISMA (Preferred Reported Items for Systematic Reviews and Meta-Analysis) recommendations, followed by meta-analysis. RESULTS: It was observed that the resistance rate ranged between 69.3-99.2% for penicillins and 30.5-97.4% for cephalosporins. The overall percentage of resistance to fluoroquinolones ranged from 4.3% to 14.0%, with the highest resistance percentage observed for levofloxacin. Resistance rates ranged between 10.7-39.8% for macrolides, 1.8-12.9% for aminoglycosides and 0.8-7.1% for tetracyclines. CONCLUSIONS: These results show that Arcobacter spp. present resistance to various antibiotics commonly used and advocate further studies of the associated resistance mechanisms.

5-Hydrazinylethylidenepyrimidines effective against multidrug-resistant Acinetobacter baumannii: Synthesis and in vitro biological evaluation of antibacterial, radical scavenging and cytotoxic activities.

Acinetobacter baumannii has emerged as an important nosocomial pathogen in recent years, with infectious outbreaks caused by multidrug-resistant strains increasing worldwide. Thus, new antibacterial treatments for multidrug-resistant A. baumannii strains are needed. In this work, a series of 5-hydrazinylethylidenepyrimidines were synthesized and in vitro evaluated against two multidrug-resistant A. baumannii strains (AcB 13/10 and AcB 73/10). Minimum inhibitory concentration results demonstrated that generally the compounds in study presented values in a low micromolar range. In the determination of in vitro bacterial growth at 24 h, it was observed that the pyrimidines 3a and 3c, with an unsubstituted hydrazinylphenyl, have bacteriostatic activity in both multidrug-resistant A. baumannii strains, with a concentration-dependent action. In general, an additive effect occurred in the combination of these compounds with gentamicin, rifampicin and polymyxin B, for both strains. Furthermore, all 5-hydrazinylethylidenepyrimidines under study presented a good 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity, generally low xanthine oxidase inhibition and low cytotoxicity in normal human dermal fibroblasts as well as potential favorable drug-likeness properties. Thus, these molecules can be considered attractive for the future development of antibacterial agents against multidrug-resistant A. baumannii.

Serum homocysteine: relationship with circulating levels of cortisol and ascorbate.

BACKGROUND/AIM: High serum levels of homocysteine and cortisol are independent risk factors for several pathologies and their levels can be regulated by some vitamins. Since the relationship between serum concentrations of homocysteine, cortisol and ascorbate has not been assessed in healthy individuals to date, it was the topic of the present work. METHODS: The study group was composed of 20 men and 40 women aged >50 years. Blood samples were collected and serum concentrations of the analytes were quantified. RESULTS: Serum homocysteine levels correlate positively with cortisol (r = 0.36, p <0.01) and age (r = 0.49, p < 0.001), and negatively with ascorbate (r = -0.30, p < 0.05) and folate (r = -0.31, p < 0.05). A negative correlation between serum levels of cortisol and ascorbate (r = -0.30, p < 0.05) was also observed. Multiple linear regression analysis showed that the best independent predictors of serum homocysteine concentration were cortisol (beta = 0.319, p < 0.003), age (beta = 0.529, p < 0.001) and folate (beta = -0.338, p < 0.001). When subjects were divided into tertiles according to their homocysteine concentration, the highest tertile of homocysteine concentration has also higher cortisol (p < 0.005) and lower ascorbate (p < 0.05) concentrations compared with the lowest tertile of homocysteine concentration. CONCLUSION: The association between serum concentrations of homocysteine, cortisol and ascorbate suggests interaction between circulating levels of these molecules.

Arcobacter butzleri Ciprofloxacin Resistance: Point Mutations in DNA Gyrase A and Role on Fitness Cost.

Arcobacter butzleri is a widely distributed emerging pathogen resistant to various classes of antimicrobial agents, namely fluoroquinolones. A. butzleri resistance to fluoroquinolones is conferred by point mutations at the antibiotic target. The aim of this study was to evaluate mutations at gyrA associated with ciprofloxacin resistance and evaluate whether acquisition of resistance impacts on fitness and stress tolerance of A. butzleri. A. butzleri ciprofloxacin mutants were generated by laboratory induction. Identification of mutations associated with ciprofloxacin resistance was performed by gyrA sequencing. Growth kinetics, cost of fitness, biofilm formation ability, and stress tolerance were assessed. Two amino acid substitutions in the quinolone resistance-determining region of GyrA were identified in the mutant strains, one previously described (Thr-85-Ile) and a new substitution (Asp-89-Tyr). No differences in growth kinetics were recorded between parental and mutant strains; however, fitness cost was variable, according to the genetic background of the strains, and independently of ciprofloxacin resistance. Overall, the ciprofloxacin resistance development did not significantly affect stress tolerance, motility, or biofilm-forming ability. In conclusion, acquisition of ciprofloxacin resistance in A. butzleri is associated with mutations in gyrA and is likely well compensated, with cost of fitness reflecting the diversity in genetic background of this bacterium.

Trisubstituted barbiturates and thiobarbiturates: Synthesis and biological evaluation as xanthine oxidase inhibitors, antioxidants, antibacterial and anti-proliferative agents.

Barbituric and thiobarbituric acid derivatives have become progressively attractive to medicinal chemists due to their wide range of biological activities. Herein, different series of 1,3,5-trisubstituted barbiturates and thiobarbiturates were prepared in moderate to excellent yields and their activity as xanthine oxidase inhibitors, antioxidants, antibacterial agents and as anti-proliferative compounds was evaluated in vitro. Interesting bioactive barbiturates were found namely, 1,3-dimethyl-5-[1-(2-phenylhydrazinyl)ethylidene]pyrimidine-2,4,6(1H,3H,5H)-trione (6c) and 1,3-dimethyl-5-[1-[2-(4-nitrophenyl)hydrazinyl]ethylidene]pyrimidine-2,4,6(1H,3H,5H)-trione (6e), which showed concomitant xanthine oxidase inhibitory effect (IC50 values of 24.3 and 27.9 µM, respectively), and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity (IC50 values of 18.8 and 23.8 µM, respectively). In addition, 5-[1-(2-phenylhydrazinyl)ethylidene]pyrimidine-2,4,6(1H,3H,5H)-trione (6d) also revealed DPPH radical scavenger effect, with an IC50 value of 20.4 µM. Moreover, relevant cytotoxicity against MCF-7 cells (IC50 = 13.3 µM) was observed with 5-[[(2-chloro-4-nitrophenyl)amino]methylene]-2-thioxodihydropyrimidine-4,6(1H,5H)-dione (7d). Finally, different 5-hydrazinylethylidenepyrimidines revealed antibacterial activity against Acinetobacter baumannii (MIC values between 12.5 and 25.0 µM) which paves the way for developing new treatments for infections caused by this Gram-negative coccobacillus bacterium, known to be an opportunistic pathogen in humans with high relevance in multidrug-resistant nosocomial infections. The most promising bioactive barbiturates were studied in silico with emphasis on compliance with the Lipinski's rule of five as well as several pharmacokinetics and toxicity parameters.

[Brown tumor of the sternum--a potential cause of false-positive result on parathyroid scintigraphy with Tc-99m sestamibi]. / Tumor marrom em esterno--uma causa potencial de resultado falso-positivo na cintilografia das paratireóides com sestamibi-99mTc.

Parathyroid scintigraphies have been used to detect pathological parathyroid glands either before as well as after the parathyroid resection surgery in patients with hyperparathyroidism. Although this test presents high specificity for detection of increased parathyroid glands, there exist causes of false positive results. In the present article, we report a case of a renal transplanted patient, with multiple lytic lesions on pelvic bones reported as brown tumors, who presented a focal uptake in the anterior portion of the superior mediastinum on Tc-99m sestamibi scintigraphy. This focal uptake, initially thought to be an ectopic parathyroid gland, after a more detailed analysis and the performance of other imaging diagnostic tests was demonstrated to be a brown tumor of the sternum.

Discordant results in Tc-99m tetrofosmin and Tc-99m sestamibi parathyroid scintigraphies.

Parathyroid scintigraphies have been used to detect pathological parathyroid glands either before as well as after the parathyroid resection surgery in patients with hyperparathyroidism. One of the most utilized techniques to perform the studies is the double-phase images with Tc-99m sestamibi, which has been shown to be very accurate in the localization of enlarged parathyroid glands. Similar to Tc-99m sestamibi, Tc-99m tetrofosmin is a radiopharmaceutical initially developed to perform myocardial perfusion study that has been used to perform parathyroid scintigraphies. Although most of the papers suggest that the overall sensitivities of both radiopharmaceuticals are similar, there are some papers questioning the accuracy of Tc-99m tetrofosmin to detect abnormal parathyroid glands. In the present article, we report a case with discordant results by both methods.