Identification of tetracycline- and erythromycin-resistant Gram-positive cocci within the fermenting microflora of an Italian dairy food product.

AIMS: Microbiological and molecular analysis of antibiotic resistance in Gram-positive cocci derived from the Italian PDO (Protected Designation of Origin) dairy food product Mozzarella di Bufala Campana. METHODS AND RESULTS: One hundred and seven coccal colonies were assigned to Enterococcus faecalis, Lactococcus lactis and Streptococcus bovis genera by ARDRA analysis (amplified ribosomal DNA restriction analysis). Among them, 16 Ent. faecalis, 26 L. lactis and 39 Strep. bovis displayed high minimum inhibitory concentration (MIC) values for tetracycline, while 17 L. lactis showed high MIC values for both tetracycline and erythromycin. Strain typing and molecular analysis of the phenotypically resistant isolates demonstrated the presence of the tet(M) gene in the tetracycline-resistant strains and of tet(S) and erm(B) in the double-resistant strains. Southern blot analysis revealed plasmid localization of L. lactis tet(M), as well as of the erm(B) and tet(S) genes. Genetic linkage of erm(B) and tet(S) was also demonstrated by PCR amplification. Conjugation experiments demonstrated horizontal transfer to Ent. faecalis strain JH2-2 only for the plasmid-borne L. lactis tet(M) gene. CONCLUSIONS: We characterized tetracycline-and erythromycin-resistance genes in coccal species, representing the fermenting microflora of a typical Italian dairy product. SIGNIFICANCE AND IMPACT OF THE STUDY: These results are of particular relevance from the food safety viewpoint, especially in the light of the potential risk of horizontal transfer of antibiotic-resistance genes among foodborne commensal bacteria.

Triamcinolone and PDT to treat exudative age-related macular degeneration and submacular hemorrhage.

PURPOSE: To evaluate the efficacy of photodynamic therapy (PDT) with verteporfin and intravitreal injection of triamcinolone to treat choroidal neovascularization (CNV) with flat sub-macular hemorrhage in age-related macular degeneration (ARMD). METHODS: A prospective, consecutive, noncomparative, interventional case series study was performed at the Instituto Oftalmologico de Alicante, Spain. Ten consecutive eyes from 10 patients with flat submacular hemorrhage secondary to ARMD were treated by PDT followed by intravitreal injection of 19.4+/-2.1 mg/0.1 mL triamcinolone 5 days later. PDT was repeated if leakage from the CNV appeared on fluorescein angiography (FA) at 3 months follow-up intervals. Main outcome measures were best-corrected visual acuity (BCVA) before and after treatment, post-treatment FA, results, and complications. RESULTS: Stable or improved BCVA was achieved in seven eyes at 6 months follow-up. Complete absence of leakage in FA was observed in five and in eight eyes at 3 and 6 months follow-up, respectively. Intraocular pressure rose in seven eyes. CONCLUSIONS: PDT followed by intravitreal triamcinolone seems useful to treat CNV with flat submacular hemorrhage in ARMD. However, further studies with longer follow-up and randomized controlled trials are necessary to assess its efficacy in the management of this difficult clinical problem.

Vascular endothelial growth factor gene polymorphisms increase the risk to develop psoriasis.

We investigated the relationship between eight polymorphisms in the gene encoding for vascular endothelial growth factor (VEGF) (-1540C > A, -1512Ins18, -1451C > T, -460T > C, -160C > T, -152G > A, -116G > A and +405G > C) and plaque-type psoriasis stratified for age at onset, gender and family history of dermatosis. For this purpose, 117 patients with chronic plaque-type psoriasis and 215 healthy subjects were enrolled. We found that being homozygous -1540AA, -1512InsIns, -1451TT, -460CC and -152AA conferred a significant risk in developing psoriasis compared with heterozygous (-1540CA, -1512 + Ins, -1451CT, -460CT and -152AG) and homozygous genotypes (-1540CC, -1512 + +-1451CC, -460TT and -152GG) grouped together [odds ratio (ORs) = 1.73, 1.73, 1.73, 1.77 and 1.87, respectively]. Conversely, having the -116AA or +405GG genotype did not significantly increase the risk of disease expression compared with other genotypes of the same loci. Interestingly, we found that -1540AA, -1512InsIns, -1451TT, -460CC and -152AA homozygous genotypes have a significant two-fold increased risk in developing psoriasis after the age of 40 years (late-onset psoriasis) (ORs = 2.19, 2.19, 2.19, 2.05 and 2.26; P = 0.02, 0.02, 0.02, 0.04 and 0.02, respectively) as compared with controls. On the contrary, we found no phenotype-genotype association of the same magnitude among the patients in whom psoriasis developed at or before the age of 40 years (early-onset psoriasis) compared with controls. Genotype distributions were not significantly different when cases and controls were stratified either by gender or family history of psoriasis. Finally, VEGF plasma concentration was not significantly different between patients and controls and was not correlated with the severity of the disease.

Phacoemulsification in the vitreous cavity for retained nuclear lens fragments.

PURPOSE: To evaluate anatomic and functional results after surgery of retained lens fragments in the vitreous cavity after previous phacoemulsification. METHODS: The authors studied retrospectively 18 patients who underwent pars plana vitrectomy (PPV) for retained nuclear lens fragments. Patients having only cortical material in the vitreous cavity were excluded. In all cases the nucleus or nuclear fragments were removed after a complete vitrectomy using perfluorocarbon injection in the vitreous cavity, associated with phacoemulsification in the vitreous cavity. The authors used a conventional phaco probe devoid of the silicone sleeve. Time lapse between cataract surgery and vitrectomy varied between 0 and 24 days (mean 8.2 +/- 7.4). Follow-up was 33.9 +/- 20.6 months (range 4 to 53). RESULTS: The mean final best-corrected visual acuity (BCVA) was 20/45 (range 20/400 to 20/20). It was 20/40 or better in 33% of patients, reaching 40% when patients with previous macular disease were excluded. A total of 61% of patients reached a final BCVA ranging from 20/50 to 20/200. Retinal detachment occurred in one eye and topical medications were necessary to manage intraocular pressure in four cases. CONCLUSIONS: PPV with intravitreous phacoemulsification is the technique of choice for dislocated nuclei or nuclear fragments in the vitreous cavity. (

[Diabetic retinopathy]. / La rétinopathie diabétique.

Diabetic retinopathy remains a serious cause of blindness in the Western World. Its major clinical signs and management are reviewed, with particular focus on advances in the diagnosis and treatment of diabetic macular edema.

[Current understanding of diabetic retinopathy]. / Principales notions actuelles sur la rétinopathie diabétique.

The incidence of diabetes continues to increase and it is estimated that the world diabetic population will have doubled in 2010. Diabetic retinopathy, one of the most frequent and precocious diabetes' complications, is increasing too and it represents a major cause of blindness in industrialized Countries. The purpose of this article is to describe the relations existing between the risk factors and the physiopathology of diabetic retinopathy, which may be helpful in taking therapeutic and prevention decisions, for the management of diabetic patients.

Inhibition of IL-2 production by Nil-2-a in murine T cells.

The loss of IL-2 production is the main defect accounting for age-related immunodeficiencies. We have investigated the molecular mechanisms involved in the decrease of IL-2 production in CD4+ T cells from aging mice. Our results demonstrate that the stability of IL-2 mRNA increases in T cells from young mice, whereas it declines in T cells from old mice with the time of stimulation, suggesting the existence of different mechanisms of post-transcriptional regulation in young and old mice. We found that the IL-2 mRNA level in T cells from young but not from old mice increased up to 6- to 10-fold by addition of cycloheximide (CHX) while the stability of IL-2 mRNA is not affected. We then looked for IL-2 inducible inhibitory factors in T cells from young and old mice and demonstrated the presence of Nil-2-a, a zinc finger protein which negatively controls IL-2 gene transcription in human cells. This protein could be detected in T cells from both young and old mice, yet, in the presence of CHX, its binding activity was reduced by 75% in T cells from young but not from old mice. These findings show that Nil-2-a accounts for the negative control of IL-2 production in the mouse and explain the reduced IL-2 production in aging.

Role of mRNA stability in the different patterns of cytokine production by CD4+ cells from young and old mice.

CD4+ cells from young (3 months) and old (19 months) mice were stimulated by plate-bound anti-CD3 monoclonal antibody (mAb) alone or also by soluble anti-CD28 mAb. Supernatants were analysed by enzyme-linked immunosorbent assay (ELISA) to determine cytokine concentrations. Total RNA was extracted from cells, reverse transcribed and the cDNA amplified by polymerase chain reaction (PCR) to evaluate the amount of specific mRNA. The results indicate that anti-CD3 alone is not sufficient to induce interleukin-2 (IL-2) production in CD4+ cells from both young and old mice. However, anti-CD28, together with anti-CD3 mAb, induces a much higher production of IL-2 in CD4+ cells from young as compared with old mice. Conversely, interferon-gamma (IFN-gamma) production is also induced by anti-CD3 alone and is higher in CD4+ cells from old as compared with young mice. Upon addition of anti-CD28 mAb, IFN-gamma production increases in both groups, but it remains much higher in old than in young mice. Also the production of IL-4 and IL-10 is induced by anti-CD3 mAb but it is increased by the addition of anti-CD28 mAb. CD4+ cells from old mice produce more IL-4 and IL-10 as compared with cells from young mice. The amounts of cytokine specific mRNA in CD4+ cells from young and old mice parallel the cytokine levels in culture supernatants. Results on the mRNA turnover indicate that when CD4+ cells are stimulated by anti-CD3 or costimulated also by anti-CD28 mAb, the IFN-gamma, IL-4 and IL-10 specific mRNAs are more stable in old than in young mice, suggesting that mRNA stability has a relevant role in the different patterns of cytokine production.

Regulation of cytokine production in aging: use of recombinant cytokines to upregulate mitogen-stimulated spleen cells.

We investigated the production of IL-2 and IFN-gamma (Th1 type) and IL-4 (Th2 type) cytokines by mitogen-activated spleen cells from young, adult and old mice. Cytokine production was evaluated in culture supernatants by CTLL proliferation (IL-2), ELISA (IFN-gamma), CT4.S proliferation (IL-4) and in mRNA extracted from activated CD4+ cells by RT-PCR (IL-2, IFN-gamma and IL-4). Results show that the production of IL-2, as protein and mRNA, is profoundly depressed by aging, whereas that of IFN-gamma, as protein and mRNA, firstly declines and then increases with age. The production of IL-4, as protein, monotonically declines with aging whereas, as mRNA, firstly decreases and then increases above the level in young mice. Spleen cells in culture were also incubated with mitogens and with a recombinant cytokine (IL-1 beta, IL-2, IL-3, IL-4, IL-12 or IFN-gamma) at various concentrations. It was found that recombinant cytokines by and large enhance cytokine production when the level induced by mitogens only is low. This conclusion applies to IL-2 and IFN-gamma production as protein and mRNA. The addition of recombinant cytokines also increases the production of IL-4 at the protein level in spleen cells from old mice but, at the mRNA level, only in spleen cells from young mice. This finding suggests age-related changes in IL-4-specific mRNA transcription rate and post-transcriptional half-life as well as translation kinetics.

[Hormonal evaluation in the diagnosis of obesity associated with "primary empty sella"]. / Valutazione ormonale nella diagnosi di obesità associata a "primary empty sella".

To determine whether obesity associated with "primary empty sella" (PES) had a characteristic hormonal pattern, we evaluated the hormonal profile in 24 simple obese and 19 obese-PES women. The latter group showed a significant delta-GH and LH impaired levels as well as plasma beta-EP significantly higher. Hence, the beta-EP measurement could be used to predict the hormonal response in these women.