Antimicrobial resistance of Salmonella serotypes isolated from slaughter-age pigs and environmental samples

The aim of this study was to determine the antimicrobial resistance patterns of Salmonella strains isolated from slaughter-age pigs and environmental samples collected at modern swine raising facilities in Brazil. Seventeen isolates of six serotypes of Salmonella enterica subsp. enterica were isolated out of 1,026 collected samples: Salmonella Typhimurium (1), Salmonella Agona (5), Salmonella Sandiego (5), Salmonella Rissen (1), Salmonella Senftenberg (4), and Salmonella Javiana (1). Resistance patterns were determined to extended-spectrum penicillin (ampicillin), broad-spectrum cephalosporins (cefotaxime and ceftriaxone), aminoglycosides (streptomycin, neomycin, gentamicin, amikacin, and tobramycin), narrow-spectrum quinolone (nalidixic acid), broad-spectrum quinolone (ciprofloxacin and norfloxacin), tetracycline, trimethoprim, and chloramphenicol. Antimicrobial resistance patterns varied among serotypes, but isolates from a single serotype consistently showed the same resistance profile. All isolates were resistant to tetracycline, streptomycin, and nalidixic acid. One isolate, Salmonella Rissen, was also resistant to cefotaxime and tobramycin. All serotypes were susceptible to ceftriaxone, norfloxacin, ciprofloxacin, ampicillin, gentamicin, and chloramphenicol. The high resistance to tetracycline and streptomycin may be linked to their common use as therapeutic drugs on the tested farms. No relation was seen between nalidixic acid and fluoroquinolone resistance.

In vitro study of the Nd : YAG laser effect on human dental enamel: Optical and scanning electron microscope analysis

Objective: the Nd:YAG laser irradiation of dental enamel was evaluated in enamel demineralization experiments in a Streptococcus mutans culture media. Summary Background Data: Previous studies had shown that a continuous wave Nd:YAG laser at an energy of approximately 67 mJ may induce an increased acid resistance in human dental enamel when exposed to severe demineralization conditions. Methods: Enamel windows of 3 x 4 cm in the buccal surface were irradiated with a continuous wave Nd:YAG laser at a wavelength of 1,064 Ecm using energy densities of from 83.75 to 187.50 J/cm(2), Enamel windows of 3 x 4 cm on the lingual surface served as control (without the laser irradiation). The enamel windows were then exposed to a Streptococcus mutans culture media at a temperature of 37 degrees C for 15 and 21 days. The laser effects and demineralization were examined both by optical microscopy and scanning electron microscopy (SEM), Results: A comparison between the lased and the unlased windows of enamel showed fusion and recrystalization of the enamel and increased acid-resistance in all groups irradiated with the Nd:YAG laser, on the other hand, the 3 x 4 delimited enamel surfaces from the control group (not irradiated with the Nd:YAG laser) showed 100% deminerization, Conclusions: These findings are consistent with the finding that laser irradiation of dental results in significant reduction of the effective solubility of enamel mineral.

Dentin bond strength: Influence of laser irradiation, acid etching, and hypermineralization

Objective: the purpose of the present study was to investigate the effects of ND:YLF laser irradiation (1.31 J/cm(2); 250 mJ per pulse), acid etching, and hypermineralization on the shear bond strength (SBS) of the Scotchbond Multi-Purpose Plus (3M Dental Products) bonding system. Summary Background Data: Previous studies had shown that the pretreatment of the dentin substrate with laser irradiation can influence the SBS, Methods: Sixty bovine incisors were selected and stored at -18 degrees C, Dentinal buccal surface was exposed and radiographs were taken to control dentin thickness, the specimens were separated into 2 groups: (1) the control, which was kept in distilled water at 4 degrees C; (2) the hypermineralized, which was kept in hypermineralizing solution at 4 degrees C for 14 days, Each group was divided into 3 subgroups according to the type of dentin pretreatment used: M (acid etching + primer + bond); AL (acid etching + primer + bond + laser); and LA (laser + acid etching + primer + bond). A standard composite resin cylinder (Z100-3M) was bonded to the dentinal surface and the SBS performed on an Instron machine (500 Kg load cell at 0.5 mm/min), followed by scanning electron microscopy (SEM) and x-ray diffraction analysis. Results: Analysis of variance (ANOVA) determined that the pretreatments influenced the SBS values (p < 0.05): AL (9.96 MPa), M (7.28 MPa), and LA (4.87 MPa), the interaction between the group and pretreatment factors also influenced the SBS (p < 0.05). The highest values were obtained for the interaction control/AL (11.64 MPa), Conclusion: the results suggested that dentin treatment with laser after the application of the adhesive system is efficient in achieving higher bond strength and is promising as a possible new adhesive substrate.

A histologic assessment of the influence of low-intensity laser therapy on wound healing in steroid-treated animals

Objective: the aim of the present study was to evaluate the effect of low-intensity laser therapy on the wound healing process treated with steroid. Background Data: Various biological effects have been associated with low-level laser therapy (LLLT). Materials and Methods: Forty-eight rats were used, and after execution of a wound on the dorsal region of each animal, they were divided into 4 groups (n = 12), receiving the following treatments: G1 (control), wounds and animals received no treatment; G2, wounds were treated with LLLT; G3, animals received an intraperitoneal injection of steroid dosage (2 mg/kg of body weight); G4, animals received steroid and wounds were treated with LLLT. The laser emission device used was a GaAIAs (904 nm), in a contact mode, with 2.75 mW gated with 2.900 Hz during 120 sec (33 J/cm(2)). After the period of 3, 7, and 14 days, the animals were sacrificed and the parts sent to histological processing and dyed using hematoxylin and eosin (HE) and Masson trichromium (MT) techniques. Results: the results have shown that the wounds treated with steroid had a delay in healing, while LLLT accelerated the wound healing process. Also, wounds treated with laser in the animals treated with steroid presented a differentiated healing process with a larger collagen deposition and also a decrease in both the inflamatory infiltrated and the delay on the wound healing process. Conclusion: LLLT accelerated healing, caused by the steroid, acting as a biostimulative coadjutant agent, balancing the undesirable effects of cortisone (in the tissue healing process.

Study of microleakage at class V cavities prepared by Er : YAG laser using rewetting surface treatment

Objective: This study was conducted to analyze microleakage in Class V cavity preparation, using rewetting (or not) just after burr or Er:YAG laser preparation of enamel and dentin walls in permanent teeth. Background Data: Several studies reported microleakage around composite restorations when cavity preparation was done or treated by Er:YAG laser. As the hybridized laser is removed when this laser is used to cut dental hard tissue, there is a need for new materials or techniques to minimize gaps and microleakage. Results: Primer solution showed significant effect in enamel and dentin, at the level of 5%, when Er:YAG laser was used as a cutting tool. Using primer solution after phosphoric acid in preparations with the laser, microleakage was similar in degree to when cavities were prepared with the burr. Conclusion: Re-wetting surface just after Er:YAG irradiation and chemical treatment with phosphoric acid using HEMA aqueous solution seems to improve the quality of bioattachment between the adhesive system and enamel/dentin, showing similarities between restoration behaviors independently of the cutting tool, whether burr or laser.

Magnesium supplementation alleviates premenstrual symptoms of fluid retention

We investigated the effect of a daily supplement of 200 mg of magnesium (as MgO) for two menstrual cycles on the severity of premenstrual symptoms in a randomized, double-blind, placebo-controlled, crossover study. A daily supplement of 200 mg of Mg (as MgO) or placebo was administered for two menstrual cycles to each volunteer, who kept a daily record of her symptoms, using a 4-point scale in a menstrual diary of 22 items. Symptoms were grouped into six categories: PMS-A (anxiety), PMS-C (craving), PMS-D (depression), PMS-H (hydration), PMS-O (other), and PMS-T (total overall symptoms). Urinary Mg output/24 hours was estimated from spot samples using the Mg/creatinine ratio. Analysis of variance for 38 women showed no effect of Mg supplementation compared with placebo in any category in the first month of supplementation. In the second month there was a greater reduction (p = 0.009) of symptoms of PMS-H (weight gain, swelling of extremities, breast tenderness, abdominal bloating) with Mg supplementation compared with placebo. Compliance to supplementation was confirmed by the greater mean estimated 24-hour urinary output of Mg (p = 0.013) during Mg supplementation (100.8 mg) compared with placebo (74.1 mg). A daily supplement of 200 mg of Mg (as MgO) reduced mild premenstrual symptoms of fluid retention in the second cycle of administration.

CYTOGENETIC EVIDENCE OF INVOLVEMENT OF CHROMOSOME REGION-15Q12 AND REGION-12Q15 IN CONDITIONS WITH ASSOCIATED OVERGROWTH

Syndromes with associated overgrowth are poorly understood. Besides their mode of inheritance, nothing is known regarding the basic genetic alterations that lead to their abnormal phenotypic manifestations. The chromosome localization of the genes involved remains unknown for this group of syndromes, with the only exception being the Wiedemann-Beckwith syndrome.

Effect of the clinical application of the GaAlAs laser in the treatment of dentine hypersensitivity

Objective: the aim of this study was to evaluate the effectiveness of the clinical use of the gallium-aluminum-arsenium (GaAlAs) laser at the maximum and minimum energies recommended by the manufacturer for the treatment of dentine hypersensitivity.Background Data: Dentine hypersensitivity (DH) is a response to a stimulus that would not usually cause pain in a healthy tooth. It is characterized by sharp pain of short duration from the denuded dentin. Its etiology is unknown. The dentin only begins to show sensitivity when exposed to the buccal environment. This exposure can result after removal of the enamel and/or dental cement, or after root denudation. Different treatments are proposed for this disorder.Materials and Methods: In this study, 25 patients, with a total number of 106 cases of DH, were treated with GaAlAs low-level laser therapy (LLLT). 65% of the teeth were premolars; 14% were incisors and molars; 6.6% were canines. The teeth were irradiated with 3 and 5 J/cm(2) for up to six sessions, with an interval of 72 It between each application, and they were evaluated initially, after each application, and at 15 and 60 days follow-up post-treatment.Results: the treatment was effective in 86.53% and 88.88% of the irradiated teeth, respectively, with the minimum and maximum energy recommended by the manufacturer. There was a statistically significant difference between DH and after a follow-up of 60 days for both groups. The difference among the energy maximum and minimum was not significant.Conclusion: the GaAlAs low-level laser was effective in reducing initial DH. A significant difference was found between initial values of hypersensitivity and after 60 days follow-up post-treatment. No significant difference was found between minimum (3 J/cm(2)) and maximum (5 J/cm(2)) applied energy.

Granulocyte colony-stimulating factor expression from transduced vascular smooth muscle cells provides sustained neutrophil increases in rats

Granulocyte colony-stimulating factor (G-CSF) regulates granulocyte precursor cell proliferation, neutrophil survival, and activation. Cyclic hematopoiesis, a disease that occurs both in humans and grey collie dogs is characterized by cyclical variations in blood neutrophils. Although the underlying molecular defect is not known, long-term daily administration of recombinant G-CSF eliminates the severe recurrent neutropenia, indicating that expression of G-CSF by gene therapy would be beneficial. As a prelude to preclinical studies in affected collie dogs, we monitored hematopoiesis in rats receiving vascular smooth muscle cells transduced to express G-CSF. Cells transduced with LrGSN, a retrovirus expressing rat G-CSF, were implanted in the carotid artery and control animals received cells transduced with LASN, a retrovirus expressing human adenosine deaminase (ADA). Test animals showed significant increases in neutrophil counts for at least 7 weeks, with mean values of 3,670 +/- 740 cells/mu l in comparison to 1,870 +/- 460 cells/mu l in controls (p < 0.001). Thus, in rats G-CSF gene transfer targeted at vascular smooth muscle cells initiated sustained production of 1,800 neutrophils/mu l, a cell number that would provide clinical benefit to patients. Lymphocytes, red cells and platelets were not different between control and test animals (p > 0.05). These studies indicate that retrovirally transduced vascular smooth muscle cells can provide sustained clinically useful levels of neutrophils in vivo.

Secretion of metalloendopeptidase 24.15 (EC 3.4.24.15)

The metalloendopeptidase EP24.15 (EC3.4.24.15) is a neuropeptide-metabolizing enzyme present in neural and endocrine tissues, presumably functioning extracellularly, Because the majority of the EP24.15 activity is identified in the soluble fraction of cellular homogenates, suggesting that the enzyme is primarily an intracellular protein, we addressed the issue of how EP24.15 arrives in the extracellular environment, We utilized a model system of neuroendocrine secretion, the AtT20 cell, According to both enzymatic activity and immunologic assays, EP24.15 was synthesized in and released from AtT20 cells. Under basal conditions and after stimulation by corticotropin-releasing hormone or the calcium ionophore A23187, EP24.15 activity accumulated in the culture medium. This secretion was not attributable to cell damage, as judged by the absence of release of cytosolic enzyme markers and the ability to exclude trypan blue dye. Pulse-chase analysis and subcellular fractionation of AtT20 cell extracts suggested that the mechanism of EP24.15 secretion is not solely via classical secretory pathways, Additionally, drugs which disrupt the classical secretory pathway, such as Brefeldin A and nocodazole, blocked A23187-stimulated EP24.15 release yet had no effect on basal EP24.15 release, suggesting differences in the basal and stimulated pathways of secretion for EP24.15. In summary, EP24.15 appears to be secreted from AtT20 pituitary cells into the extracellular milieu, where the enzyme can participate in the physiologic metabolism of neuropeptides.

Clinical comparison between conventional suture and vaporization with carbon dioxide laser in rat's skin

Objective: This study compares wound healing efficiency on a rat's skin when the incision was closed with a conventional suture versus vaporized with a CO2 laser. Materials and Methods: In this study, 24 rats were used, and two longitudinal incisions were made with a conventional scalpel in the dorsum of each rat. The left incision was sutured with nylon thread, and the right incision was closed by vaporization with a defocused CO2 laser in continuous mode with an 8-watt power density. Clinical photographs were taken immediately after the procedure, 24 h later, and after 3, 7, 14, and 21 days, documenting the healing of the incision. Results: the results showed that there was an initial delay in wound repair in the vaporized incision as compared to the scalpel incision, but after 21 days, both incisions showed the same clinical characteristics. However, the vaporized incision showed no trauma of the tissue, as opposed to the sutured incision, and no hemorrhagic complications. Conclusion: These results suggest that the CO2 laser can eventually replace the use of sutures.

Monoclonal antibodies against the 43,000 da glycoprotein from Paracoccidioides brasiliensis modulate laminin-mediated fungal adhesion to epithelial cells and pathogenesis

The surface glycoprotein gp43, a highly immunogenic component of Paracoccidioides brasiliensis, is used in the serodiagnosis of paracoccidioidomycosis (PCM) and has recently been shown to specifically bind the extracellular matrix protein laminin, Binding to laminin induces the increased adhesion of the fungus to epithelial cells; a hamster testicle infection model has shown that the gp43-dependent binding of fungal cells to laminin enhances their pathogenicity in vivo. We report on the production and characterization of 12 monoclonal antibodies against the gp43 that recognize peptide sequences in the molecule detecting at least three different epitopes as well as different isoforms of this antigen. MAbs interfered in the fungal pathogenicity in vivo either by inhibiting or enhancing granuloma formation and tissue destruction, Results suggest that P. brasiliensis propagules may start infection in man by strongly adhering to human lung cells, Thus, laminin-mediated fungal adhesion to human lung carcinoma (A549) cells was much more intense than to Madin-Darby canine kidney cells (MDCK), indicating differences in binding affinity, Subsequent growth of fungi bound to the lung cells could induce the granulomatous inflammatory reaction characteristic of PCM. Both steps are greatly stimulated by laminin binding in infective cells expressing gp43.

Serological evidence of Infection by Leishmania (Leishmania) infantum (Synonym: Leishmania (Leishmania) chagasi) in free-ranging wild mammals in a nonendemic region of the state of São Paulo, Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

In Vitro and In Vivo Validation of Human and Goat Chondrocyte Labeling by Green Fluorescent Protein Lentivirus Transduction

We investigated whether human articular chondrocytes can be labeled efficiently and for long-term with a green fluorescent protein (GFP) lentivirus and whether the viral transduction would influence cell proliferation and tissue-forming capacity. The method was then applied to track goat articular chondrocytes after autologous implantation in cartilage defects. Expression of GFP in transduced chondrocytes was detected cytofluorimetrically and immunohistochemically. Chondrogenic capacity of chondrocytes was assessed by Safranin-O staining, immunostaining for type II collagen, and glycosaminoglycan content. Human articular chondrocytes were efficiently transduced with GFP lentivirus (73.4 +/- 0.5% at passage 1) and maintained the expression of GFP up to 22 weeks of in vitro culture after transduction. Upon implantation in nude mice, 12 weeks after transduction, the percentage of labeled cells (73.6 +/- 3.3%) was similar to the initial one. Importantly, viral transduction of chondrocytes did not affect the cell proliferation rate, chondrogenic differentiation, or tissue-forming capacity, either in vitro or in vivo. Goat articular chondrocytes were also efficiently transduced with GFP lentivirus (78.3 +/- 3.2%) and maintained the expression of GFP in the reparative tissue after orthotopic implantation. This study demonstrates the feasibility of efficient and relatively long-term labeling of human chondrocytes for co-culture on integration studies, and indicates the potential of this stable labeling technique for tracking animal chondrocytes for in cartilage repair studies.

Toward cell therapy using placenta-derived cells: disease mechanisms, cell biology, preclinical studies, and regulatory aspects at the round table

Among the many cell types that may prove useful to regenerative medicine, mounting evidence suggests that human term placenta-derived cells will join the list of significant contributors. In making new cell therapy-based strategies a clinical reality, it is fundamental that no a priori claims are made regarding which cell source is preferable for a particular therapeutic application. Rather, ongoing comparisons of the potentiality and characteristics of cells from different sources should be made to promote constant improvement in cell therapies, and such comparisons will likely show that individually tailored cells can address disease-specific clinical needs. The principle underlying such an approach is resistance to the notion that comprehensive characterization of any cell type has been achieved, neither in terms of phenotype nor risks-to-benefits ratio. Tailoring cell therapy approaches to specific conditions also requires an understanding of basic disease mechanisms and close collaboration between translational researchers and clinicians, to identify current needs and shortcomings in existing treatments. To this end, the international workshop entitled "Placenta-derived stem cells for treatment of inflammatory diseases: moving toward clinical application" was held in Brescia, Italy, in March 2009, and aimed to harness an understanding of basic inflammatory mechanisms inherent in human diseases with updated findings regarding biological and therapeutic properties of human placenta-derived cells, with particular emphasis on their potential for treating inflammatory diseases. Finally, steps required to allow their future clinical application according to regulatory aspects including good manufacturing practice (GMP) were also considered. In September 2009, the International Placenta Stem Cell Society (IPLASS) was founded to help strengthen the research network in this field.