Antiviral Profiling of C-18- or C-19-Functionalized Semisynthetic Abietane Diterpenoids.

Viral infections affect several million patients annually. Although hundreds of viruses are known to be pathogenic, only a few can be treated in the clinic with available antiviral drugs. Naturally based pharmacotherapy may be a proper alternative for treating viral diseases. Several natural and semisynthetic abietane-type diterpenoids have shown important antiviral activities. In this study, a biological evaluation of a number of either C-18- or C-19-functionalized known semisynthetic abietanes against Zika virus, Dengue virus, Herpes virus simplex type 1, and Chikungunya virus are reported. Semisynthetic abietane ferruginol and its analogue 18-(phthalimid-2-yl)ferruginol displayed broad-spectrum antiviral properties. The scale-up synthesis of this analogue has been optimized for further studies and development. This molecule displayed an EC50 between 5.0 and 10.0 µM against Colombian Zika virus strains and EC50 = 9.8 µM against Chikungunya virus. Knowing that this ferruginol analogue is also active against Dengue virus type 2 (EC50 = 1.4 µM, DENV-2), we can conclude that this compound is a promising broad-spectrum antiviral agent paving the way for the development of novel antivirals.

Dual modification of achira (Canna indica L) starch and the effect on its physicochemical properties for possible food applications.

The aim of this study was to evaluate the effect of acid hydrolysis and succination upon single and a combination of both of them as a dual modification on the morphological, structural, thermal, and pasting profile of the achira starch in order to expand its potential food applications. The surface of achira starch granules was eroded with acid hydrolysis, while the succination resulted in the formation of pores or cavities, having a slight impact on the crystallinity and the gelatinization enthalpy. Succinated starch presented the lowest transition temperatures (To = 60.29 °C, Tp = 65.03 °C and Te = 69.86 °C) compared to other starches in this study. The succination increased the final viscosity (3808 cp) when compared with the native starch (3114 cp), while acid hydrolysis resulted in a decreased value (735 cp). These are desirable properties for its possible use as an additive in bakery industry processes.

Ectopic Papillary Thyroid Cancer with Distant Metastasis.

Ectopic thyroid tissue is a rare clinical entity wherein malignant lesions may arise, the most common one being papillary carcinoma. We present a case of a 68-year-old female who presented with a growing mass in the right clavicle. An MR of the shoulder showed a soft tissue mass arising from the anterior margin of the right distal clavicle. A fine needle aspiration of the mass showed papillary thyroid carcinoma. PET/CT confirmed the clavicular and mediastinal mass. Excision of the clavicular mass and total thyroidectomy with modified right neck dissection were performed. Pathology revealed no evidence of malignancy in the thyroid; lymph nodes were positive for metastatic thyroid carcinoma. Postsurgery CT showed the superior mediastinal mass with surrounding adenopathy; radioiodine (RAI) treatment with dose of 142.1 mCi [5257.7 MBq] was recommended. Posttherapy whole-body scan (WBS) showed RAI avid tissue in the neck and superior mediastinum. Follow-up chest CT revealed pulmonary nodules that increased in number and size; a second RAI therapeutic dose was given. The posttherapy RAI WBS was negative. Repeat PET/CT showed multiple hypermetabolic lesions in the mediastinum, neck, lymph nodes, pulmonary nodes, and right shoulder. The FDG avid lesions with no RAI uptake suggested tumor dedifferentiation.

Attachment of 13 Types of Foodborne Bacteria to Jalapeño and Serrano Peppers and Antibacterial Effect of Roselle Calyx Extracts, Sodium Hypochlorite, Colloidal Silver, and Acetic Acid against These Foodborne Bacteria on Peppers.

Chili peppers are a very important crop in Mexico. However, these peppers have been associated with Salmonella infection outbreaks in the United States, and Salmonella and diarrheagenic Escherichia coli pathotypes have been isolated from jalapeño and serrano peppers in Mexico. To decrease microbial contamination of fruits and vegetables, chemical agents are commonly used; however, chemical agents used to eliminate pathogenic bacteria on vegetables have a limited antimicrobial effect. Roselle ( Hibiscus sabdariffa ) calyces have been reported to have an antimicrobial effect on pathogenic bacteria. In the present study, the antibacterial effect of four roselle calyx extracts (water, methanol, acetone, and ethyl acetate), sodium hypochlorite, colloidal silver, and acetic acid against foodborne bacteria was evaluated on contaminated jalapeño and serrano peppers. The 13 types of foodborne bacteria evaluated were Listeria monocytogenes , Shigella flexneri , Salmonella Typhimurium, Salmonella Typhi, Salmonella Montevideo, Staphylococcus aureus , E. coli O157:H7, five E. coli pathotypes (Shiga toxin producing, enteropathogenic, enterotoxigenic, enteroinvasive, and enteroaggregative), and Vibrio cholerae O1. All 13 types attached to both pepper types, with no significant differences in attachment between jalapeño and serrano peppers. Roselle calyx extract treatment resulted in a greater reduction in levels of all foodborne bacteria than did treatment with sodium hypochlorite, colloidal silver, and acetic acid on both pepper types. Roselle calyx extracts may be a useful for disinfection of chili peppers in the field, processing plants, restaurants, and homes.

Nanocomposites of rice and banana flours blend with montmorillonite: partial characterization.

Rice and banana flours are inexpensive starchy materials that can form films with more improved properties than those made with their starch because flour and starch present different hydrophobicity. Montmorillonite (MMT) can be used to further improve the properties of starch-based films, which has not received much research attention for starchy flours. The aim of this work was to evaluate the mechanical and barrier properties of nanocomposite films of banana and rice flours as matrix material with addition of MMT as a nanofiller. MMT was modified using citric acid to produce intercalated structures, as verified by the X-ray diffraction pattern. The intercalated MMT was blended with flour slurries, and films were prepared by casting. Nanocomposite films of banana and rice flours presented an increase in the tensile at break and elongation percentage, respectively, more than their respective control films without MMT. This study showed that banana and rice flours could be alternative raw materials to use in making nanocomposite films.

Cold storage of human milk: effect on its bacterial composition.

OBJECTIVES: In the last few years, it has been proved that human milk contains bacteria that constitute an important factor in the initiation and development of the neonatal gut microbiota. In this context, the objective of this study was to evaluate the effect of cold storage on the natural bacterial composition of breast milk. MATERIALS AND METHODS: Breast milk samples provided by 34 healthy women and collected either by manual expression (n = 27) or breast pump (n = 7), were plated onto several culture media immediately after arrival at the laboratory (day 0) and after storage at -20 degrees C for 6 weeks. A high number of isolates from 8 of the women were identified at the species level. RESULTS: No statistically significant differences were observed between the counts obtained at both sampling times in those media in which growth was detected. In all of the culture media, bacterial counts in pump-collected samples were higher than in those obtained by manual expression. Staphylococci and streptococci were the predominant bacteria in both fresh and frozen samples, Staphylococcus epidermidis being the most abundant species at both sampling times. Lactic acid bacteria and bifidobacteria were also present in fresh and frozen breast milk samples, but among them, only 1 species (Lactobacillus gasseri) could be isolated at both sampling times. CONCLUSIONS: The results of this study suggest that cold storage of milk at -20 degrees C for 6 weeks does not significantly affect either the quantitative or the qualitative bacterial composition of breast milk.

Staphylococcus epidermidis strains isolated from breast milk of women suffering infectious mastitis: potential virulence traits and resistance to antibiotics.

BACKGROUND: Although Staphylococcus aureus is considered the main etiological agent of infectious mastitis, recent studies have suggested that coagulase-negative staphylococci (CNS) may also play an important role in such infections. The aims of this work were to isolate staphylococci from milk of women with lactational mastitis, to select and characterize the CNS isolates, and to compare such properties with those displayed by CNS strains isolated from milk of healthy women. RESULTS: The milk of 30 women was collected and bacterial growth was noted in 27 of them, of which Staphylococcus epidermidis was isolated from 26 patients and S. aureus from 8. Among the 270 staphylococcal isolates recovered from milk of women with mastitis, 200 were identified as Staphylococcus epidermidis by phenotypic assays, species-specific PCR and PCR sequencing. They were typified by pulsed field gel electrophoresis (PFGE) genotyping. The PFGE profiles of the S. epidermidis strains were compared with those of 105 isolates from milk of healthy women. A representative of the 76 different PFGE profiles was selected to study the incidence of virulence factors and antibiotic resistance. The number of strains that contained the biofilm-related icaD gene and that showed resistance to oxacillin, erythromycin, clindamycin and mupirocin was significantly higher among the strains isolated from mastitic milk. CONCLUSION: S. epidermidis may be a frequent but largely underrated cause of infectious mastitis in lactating women. The resistance to diverse antibiotics and a higher ability to form biofilms found among the strains isolated from milk of women suffering mastitis may explain the chronic and/or recurrent nature of this infectious condition.

Isolation of bifidobacteria from breast milk and assessment of the bifidobacterial population by PCR-denaturing gradient gel electrophoresis and quantitative real-time PCR.

The objective of this work was to elucidate if breast milk contains bifidobacteria and whether they can be transmitted to the infant gut through breastfeeding. Twenty-three women and their respective infants provided samples of breast milk and feces, respectively, at days 4 to 7 after birth. Gram-positive and catalase-negative isolates from specific media with typical bifidobacterial shapes were identified to the genus level by F6PPK (fructose-6-phosphate phosphoketolase) assays and to the species level by 16S rRNA gene sequencing. Bifidobacterial communities in breast milk were assessed by PCR-denaturing gradient gel electrophoresis (PCR-DGGE), and their levels were estimated by quantitative real-time PCR (qRTi-PCR). Bifidobacteria were present in 8 milk samples and 21 fecal samples. Bifidobacterium breve, B. adolescentis, and B. bifidum were isolated from milk samples, while infant feces also contained B. longum and B. pseudocatenulatum. PCR-DGGE revealed the presence of one to four dominant bifidobacterial bands in 22 milk samples. Sequences with similarities above 98% were identified as Bifidobacterium breve, B. adolescentis, B. longum, B. bifidum, and B. dentium. Bifidobacterial DNA was detected by qRTi-PCR in the same 22 milk samples at a range between 40 and 10,000 16S rRNA gene copies per ml. In conclusion, human milk seems to be a source of living bifidobacteria for the infant gut.

Is meconium from healthy newborns actually sterile?

In a previous study, bacteria were able to be isolated from umbilical cord blood of healthy neonates and from murine amniotic fluid obtained by caesarean section. This suggested that term fetuses are not completely sterile and that a prenatal mother-to-child efflux of commensal bacteria may exist. Therefore, the presence of such bacteria in meconium of 21 healthy neonates was investigated. The identified isolates belonged predominantly to the genuses Enterococcus and Staphylococcus. Later, a group of pregnant mice were orally inoculated with a genetically labelled E. fecium strain previously isolated from breast milk of a healthy woman. The labelled strain could be isolated and PCR-detected from meconium of the inoculated animals obtained by caesarean section one day before the predicted date of labor. In contrast, it could not be detected in samples obtained from a non-inoculated control group.

Isolation of commensal bacteria from umbilical cord blood of healthy neonates born by cesarean section.

In a previous study, lactic acid bacteria were isolated from meconium obtained from healthy neonates born by cesarean section. Such a finding suggested that term fetuses are not completely sterile, and that a mother-to-child efflux of commensal bacteria may exist. Therefore, presence of such bacteria in umbilical cord blood of healthy neonates born by elective cesarean section was investigated. The blood samples were submitted to an enrichment step and then inoculated onto agar plates. All the identified isolates belonged to the genus Enterococcus, Streptococcus, Staphylococcus, or Propionibacterium. Later, a group of pregnant mice were orally inoculated with a genetically labeled E. faecium strain previously isolated from breast milk of a healthy woman. The labeled strain could be isolated and polymerase chain reaction detected from the amniotic fluid of the inoculated animals. In contrast, it could not be detected in the samples obtained from a noninoculated control group.

Probiotic potential of 3 Lactobacilli strains isolated from breast milk.

Breast milk is an important factor in the initiation, development, and composition of the neonatal gut microbiota. In a previous study, the authors isolated lactic acid bacteria from milk of healthy mothers. Since some of the identified isolates belonged to the genus Lactobacillus, the objective of this work was to evaluate the probiotic potential of 2 Lactobacillus gasseri and 1 Lactobacillus fermentum strains. Different assays, including survival to conditions simulating those existing in the gastrointestinal tract, production of antimicrobial compounds, adherence to intestinal cells, production of biogenic amines, degradation of mucin, enzymatic profile, and pattern of antibiotic resistance, were performed. Globally, the results showed that the probiotic potential of lactobacilli isolated from milk of healthy mothers is, at least, similar to that of the strains commonly used in commercial probiotic products. This fact, together with the presence of prebiotic substances, indicates that breast milk is a natural synbiotic food.

Human milk is a source of lactic acid bacteria for the infant gut.

OBJECTIVES: To investigate whether human breast milk contains potentially probiotic lactic acid bacteria, and therefore, whether it can be considered a synbiotic food. Study design Lactic acid bacteria were isolated from milk, mammary areola, and breast skin of eight healthy mothers and oral swabs and feces of their respective breast-fed infants. Some isolates (178 from each mother and newborn pair) were randomly selected and submitted to randomly amplified polymorphic DNA (RAPD) polymerase chain reaction analysis, and those that displayed identical RAPD patterns were identified by 16S rDNA sequencing. RESULTS: Within each mother and newborn pair, some rod-shaped lactic acid bacteria isolated from mammary areola, breast milk, and infant oral swabs and feces displayed identical RAPD profiles. All of them, independently from the mother and child pair, were identified as Lactobacillus gasseri. Similarly, among coccoid lactic acid bacteria from these different sources, some shared an identical RAPD pattern and were identified as Enterococcus faecium. In contrast, none of the lactic acid bacteria isolated from breast skin shared RAPD profiles with lactic acid bacteria of the other sources. CONCLUSIONS: Breast-feeding can be a significant source of lactic acid bacteria to the infant gut. Lactic acid bacteria present in milk may have an endogenous origin and may not be the result of contamination from the surrounding breast skin.

Effects of Lactobacillus spp. on Cytokine Production by RAW 264.7 Macrophage and EL-4 Thymoma Cell Lines.

We have hypothesized that lactobacilli used in fermented dairy products can stimulate immune function via enhancing cytokine secretion by leukocytes. To test the effects of lactobacilli on cytokine production, RAW 264.7 cells (macrophage model) and EL4.IL-2 thymoma cells (T helper cell model) were cultured in the presence of 16 representative strains of heat-killed Lactobacillus spp. cells. In unstimulated RAW 264.7 cells, most lactobacilli, when present at concentrations from 106 to 108 bacterial cells per ml, induced marked tumor necrosis factor alpha (TNF-α) production (up to 411-fold) compared to the negligible TNF-α in controls. A strain-dependent increase in interleukin 6 (IL-6) production (up to 88-fold) was also observed without stimulation at concentrations of 108 bacteria per ml. Upon concurrent stimulation of RAW 264.7 cells with lipopolysaccharide, both IL-6 and TNF-α production were enhanced between 4.2- and 10.6-fold and 1.8- and 8.7-fold, respectively, when cultured with 108 lactobacilli per ml. In unstimulated EL4.IL-2 cells, lactobacilli had no effect on interleukin 2 (IL-2) or interleukin 5 (IL-5) production. Upon stimulation of EL4.IL-2 cells with phorbol 12-myristate-13-acetate, IL-2 secretion increased up to 1.9-fold in the presence of 106 L. bulgaricus Lr 79 cells per ml whereas this cytokine decreased in the presence of 107 or 108 lactobacilli per ml. In contrast, IL-5 secretion increased in the presence of increasing concentrations of lactobacilli (up to 3.1-fold with 108 L. bulgaricus NCK 231 cells per ml). The results indicated that direct interaction of most lactobacilli with macrophages resulted in a concentration-dependent enhancement of cytokine production, whereas the effects on cytokine production by the T-cell model were smaller and strain dependent. The in vitro approaches employed here should be useful in further characterization of the effects of lactobacilli on the gut and systemic immune systems.