Detection of linezolid resistance due to the optrA gene in Enterococcus faecalis from poultry meat from the American continent (Colombia).

Background: Three Enterococcus isolates obtained from retail chicken collected in 2010-11 as part of the Colombian Integrated Program for Antimicrobial Resistance Surveillance (COIPARS) showed reduced susceptibility towards linezolid (MIC 8 mg/L). Objectives: This study aimed at characterizing the isolates resistant to linezolid and detecting the resistance mechanism. Methods: Strains were analysed in 2011-12 without successful detection of the resistance mechanism. All isolates were found negative for the cfr gene and no 23S rRNA mutations were detected. In 2016, with the novel resistance gene optrA being described, the WGS data were re-analysed using in silico genomic tools for confirmation of species, detection of virulence and resistance genes, MLST and SNP analyses and comparison of the genetic environment with the previously published plasmid pE349. Results: : Three Enterococcus faecalis isolates were found positive for the optrA gene encoding resistance to linezolid and phenicols. Additional screening of 37 enterococci strains from the same study did not detect any further positives. Typing showed that two of the isolates belong to ST59, while the last belongs to ST489. All isolates carry genes encoding resistance to macrolide-lincosamide-streptogramin B, tetracycline and phenicols. In addition, the ST489 isolate also carries genes conferring aminoglycoside resistance and is resistant to quinolones, but no plasmid-mediated gene was detected. The optrA gene regions of the three plasmids showed high similarity to the originally reported optrA -carrying plasmid pE349. Conclusions: To the best of our knowledge, this is the first description of the optrA gene in E. faecalis isolated from poultry meat in the Americas.

Roxithromycin reduces cyclosporine-induced gingival hyperplasia in renal transplant patients.

Gingival overgrowth (GO) is a common side effect of chronic cyclosporine use. The average prevalence of GO is about 30%, ranging from 10% to 85% in various series, due to diverse aggravating risk factors: drug interactions with calcium channel blockers, age, cyclosporine dose, bacterial plaque, and genetic predisposition. Recent studies have demonstrated elevated levels of specific cytokines particularly transforming growth factor-beta (TGF-beta) in hyperplastic gingival tissue, suggesting that this growth factor plays a role in the accumulation of the extracellular matrix. Until recently treatment for this complication was only surgical. Nowadays, several studies have been performed to evaluate the effects of antibiotic treatment on the regression of GO. In the present study, we used roxithromycin, a macrolide antibiotic that has inhibitory effect on TGF-beta production by inflammatory cells. The results suggested that roxithromycin may be an important therapeutic tool to reduce cyclosporine-induced GO.

Observing bacterial activity interferometrically.

It is shown that bacterial activity, even of slowly growing species, can be detected by precise interferometric measurements of refractive index changes of the culture medium. The bacteria-containing sample is kept in an isothermal block together with a reference liquid without bacteria. The biological activity is obtained from the difference of the index changes of these samples. Experiments were performed with Bacilo Calmette-Guérin. The order of magnitude of the observed total refractive index change was compatible with theoretical estimates based on the amount of available oxygen. An unexpected positive index change during the lag phase was observed, which might permit fast diagnostics in medical applications. This technique may provide cheap and quick tests of bacterial susceptibility with respect to antibiotics.

Oral manifestation of sporotrichosis in AIDS patients.

A wide spectrum of oral lesions has been associated with human immunodeficiency viral infection (HIV), or AIDS. This report describes the case of an HIV-infected patient who developed a case of disseminated sporotrichosis whose first clinical sign was the presence of orofacial lesions. A histopathological study of this patient's biopsy specimens taken from the oropharyngeal lesions revealed a number of rounded and/or oval free-spore forms of Sporothrix schenkii, the identification of which was corroborated by culturing skin lesion exudate on Sabouraud's glucose agar. To the best of our knowledge to date, this is the first time a case of the oral manifestation of sporotrichosis in association with HIV infection has been described in the dental literature.

Experimental Mycobacterium leprae infection in BALB/c mice: effect of BCG administration on TNF-alpha production and granuloma development.

In the present study, the experimental model of Mycobacterium leprae infection in the foot pads of BALB/c mice was used to investigate the effects of BCG administration on tumor necrosis factor-alpha (TNF-alpha) production and granuloma development. It was observed that mice intravenously infected with BCG 7 months after M. leprae inoculation into the foot pads presented a more effective mycobacteria clearance, revealed by a significant reduction of BCG-colony forming units in the spleen and by the reduction of acid-fast bacilli (AFB) in the foot pads. BCG infection at the peak of M. leprae infection also modulated the granulomatous response to M. leprae by converting mononuclear granulomas into an epithelioid-cell granuloma. Furthermore, lower TNF-alpha serum levels were detected in M. leprae-infected mice when compared to mice infected with M. leprae + BCG. An analysis of the TNF-alpha gene expression in the spleen by semiquantitative reverse transcription-polymerase chain reactions (RT-PCR) demonstrated that co-infection with BCG induced an earlier expression of TNF-alpha mRNA than in M. leprae-infected mice. The numbers of TNF-alpha-positive cells and apoptotic cells were also enhanced in epithelioid versus non-epithelioid granulomas. As a whole, the data suggest that co-infection of M. leprae-infected mice with BCG modulates TNF-alpha synthesis which, in turn, leads to induction of protective epithelioid granuloma formation in the foot pads and subsequent mycobacterial clearance. Macrophage differentiation into epithelioid cells, in association with the enhancement of TNF-alpha production at the granuloma site, may represent a triggering signal that induced apoptosis in these cells, leading to mycobacterial elimination. Moreover, the rate of apoptosis in epithelioid granulomas may well be related to the extent of immunopathologically mediated tissue damage.

Effect of the injection of an extract of Ascaris suum on macrophage activation during the early phase of Mycobacterium bovis BCG infection in C57Bl/6 mice.

Injection of an Ascaris suum extract (Asc) affects both the humoral and cellular immune responses to unrelated antigens when it is co-administered with these antigens. In the present study we evaluated the effect of Asc on macrophage activation in the early phase of Mycobacterium bovis BCG (Pasteur strain TMCC 1173) infection in C57Bl/6 mice. C57Bl/6 mice were injected intraperitoneally (ip) with 0.1 mg BCG (BCG group) or BCG plus 1 mg Asc (BCG + Asc group). The peritoneal exudates were obtained at 2, 7 and 14 days after infection. The numbers of IFN-gamma-secreting cells were assessed by the ELISPOT assay. Nitric oxide (NO) production was measured by the Griess method and by the evaluation of NADPH diaphorase activity in the peritoneal exudates. The administration of Asc extract increased NADPH diaphorase activity (2 days: control = 0, BCG = 7%, BCG + Asc = 13%, and Asc = 4%; 7 days: control = 4, BCG = 13%, BCG + Asc = 21%, and Asc = 4.5%) and TNF-alpha levels (mean +/- SD; 2 days: control = 0, BCG = 169 +/- 13, BCG + Asc = 202 +/- 37, and Asc = 0; 7 days: control = 0, BCG = 545 +/- 15.5, BCG + Asc = 2206 +/- 160.6, and Asc = 126 +/- 26; 14 days: control = 10 +/- 1.45, BCG = 9 +/- 1.15, BCG + Asc = 126 +/- 18, and Asc = 880 +/- 47.67 pg/ml) in the early phase of BCG infection. Low levels of NO production were detected at 2 and 7 days after BCG infection, increasing at 14 days (mean +/- SD; 2 days: control = 0, BCG = 3.7 +/- 1.59, BCG + Asc = 0.82 +/- 0.005, Asc = 0.48 +/- 0.33; 7 days: control = 0, BCG = 2.78 +/- 1.54, BCG + Asc = 3.07 +/- 1.05, Asc = 0; 14 days: control = 0, BCG = 9.05 +/- 0.53, BCG + Asc = 9.61 +/- 0.81, Asc = 10.5 +/- 0.2 (2 x 10(6)) cells/ml). Furthermore, we also observed that Asc co-injection induced a decrease of BCG-colony-forming units (CFU) in the spleens of BCG-infected mice during the first week of infection (mean +/- SD; 2 days: BCG = 1.13 +/- 0.07 and BCG + Asc = 0.798 +/- 0.305; 7 days: BCG = 1.375 +/- 0. 194 and BCG + Asc = 0.548 +/- 0.0226; 14 days: BCG = 0.473 +/- 0.184 and BCG + Asc = 0.675 +/- 0.065 (x 10(2)) CFU). The present data suggest that Asc induces the enhancement of the immune response in the early phase of BCG infection.

Effect of the injection of an extract of Ascaris suum on macrophage activation during the early phase of Mycobacterium bovis BCG infection in C57Bl/6 mice

Injection of an Ascaris suum extract (Asc) affects both the humoral and cellular immune responses to unrelated antigens when it is co-administered with these antigens. In the present study we evaluated the effect of Asc on macrophage activation in the early phase of Mycobacterium bovis BCG (Pasteur strain TMCC 1173) infection in C57Bl/6 mice. C57Bl/6 mice were injected intraperitoneally (ip) with 0.1 mg BCG (BCG group) or BCG plus 1 mg Asc (BCG + Asc group). The peritoneal exudates were obtained at 2, 7 and 14 days after infection. The numbers of IFN-g-secreting cells were assessed by the ELISPOT assay. Nitric oxide (NO) production was measured by the Griess method and by the evaluation of NADPH diaphorase activity in the peritoneal exudates. The administration of Asc extract increased NADPH diaphorase activity (2 days: control = 0, BCG = 7 per cent, BCG + Asc = 13 per cent, and Asc = 4 per cent; 7 days: control = 4, BCG = 13 per cent, BCG + Asc = 21 per cent, and Asc = 4.5 per cent) and TNF-a levels (mean + or - SD; 2 days: control = 0, BCG = 169 + or - 13, BCG + Asc = 202 + or - 37, and Asc = 0; 7 days: control = 0, BCG = 545 + or - 15.5, BCG + Asc = 2206 + or - 160.6, and Asc = 126 + or - 26; 14 days: control = 10 + or - 1.45, BCG = 9 + or - 1.15, BCG + Asc = 126 + or - 18, and Asc = 880 + or - 47.67 pg/ml) in the early phase of BCG infection. Low levels of NO production were detected at 2 and 7 days after BCG infection, increasing at 14 days (mean + or - SD; 2 days: control = 0, BCG = 3.7 + or - 1.59, BCG + Asc = 0.82 + or - 0.005, Asc = 0.48 + or - 0.33; 7 days: control = 0, BCG = 2.78 + or - 1.54, BCG + Asc = 3.07 + or - 1.05, Asc = 0; 14 days: control = 0, BCG = 9.05 + or - 0.53, BCG + Asc = 9.61 + or - 0.81, Asc = 10.5 + or - 0.2 (2 x 106) cells/ml). Furthermore, we also observed that Asc co-injection induced a decrease of BCG-colony-forming units (CFU) in the spleens of BCG-infected mice during the first week of infection (mean + or - SD; 2 days: BCG = 1.13 + or - 0.07 and BCG + Asc = 0.798 + or - 0.305; 7 days: BCG = 1.375 + or - 0.194 and BCG + Asc = 0.548 + or - 0.0226; 14 days: BCG = 0.473 + or - 0.184 and BCG + Asc = 0.675 + or - 0.065 (x 102) CFU). The present data suggest that Asc induces the enhancement of the immune response in the early phase of BCG infection.

Oral manifestation of tertiary syphilis: case report.

This paper describes a case of benign tertiary syphilis represented by a solitary hypertrophic lesion on the dorsum surface of the tongue. The diagnosis was confirmed by serologic tests (VDRL and FTA-ABS). Histopathological analysis of biopsy specimens revealed, in the lamina propria, the presence of well-developed granulomas associated with necrotic areas (gummatous lesion). Currently, tertiary syphilis is rarely seen; however, this case emphasizes that it still exists and must be considered in the differential diagnosis of inflammatory oral lesions.

The effect of thalidomide on BCG-induced granulomas in mice.

Granuloma proliferation is the result of a series of complex biological events in which a variety of cell types and cytokines are involved. Tumor necrosis factor alpha (TNF-alpha) plays a central role. In the present study, we investigated the effect of thalidomide (alpha-N-pthalimidoglutarimide), a selective inhibitor of TNF-alpha synthesis, on granuloma formation during BCG infection in Oncins France 1 (OF-1) mice. Subcutaneous injections of 30 mg/kg body weight of thalidomide daily for 14, 21 or 28 days into the mice resulted in the reduction of the size and total number of liver granulomas. The most striking effect of thalidomide was observed after 28 days, when the total number of liver granulomas was reduced by as much as 40% (P < 0.05). Serum TNF-alpha levels of thalidomide-treated mice were significantly lower (85%) than control mice on day 14 and remained lower (55%) on days 21 and 28. Positive immunohistochemical staining specific for TNF-alpha were demonstrable only in well-developed granulomas in which central mononuclear cells presented extensive differentiation into epithelioid cells. Daily administration of thalidomide for 21 to 28 days to the BCG-infected mice inhibited local TNF-alpha expression in well-developed granulomas. The mechanisms by which thalidomide modulates the granuloma proliferation are discussed.

The effect of thalidomide on BCG-induced granulomas in mice

Granuloma proliferation is the result of a series of complex biological events in wich a variety of cell types and cytokines are involved. Tumor necrosis factor alpha (TNF-Ó) plays a central role. In the present study, we investigated the effect of thalidomide (Ó-N-pthalimidoglutarimide), a sective inhibitor of TNF-Ó synthesis, on granuloma formation during BCG infection in Oncins France 1 (OF-1) mice. Subcutaneous injections of 30 mg/kg body weight of thalidomide daily for 14,21 or 28 days into the mice resulted in the reduction of the size and total number of liver granulomas. The most strikinf effect of thalidomide was observed after 28 days, when the total number of liver granulomas was reduced by as much as 40 percent (P<0.05). Serum TNF-Ó levels of thalidomide-treatment mice were significantly lower (85 percent) than control mice on day 14 and remained lower...