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The spread of infections from multi-resistant bacteria in hospitals around the world is raising at an alarming rate. With the increasing capacity of bacteria to develop resistance to traditional antibiotics that target a particular metabolic pathway inside the cell, the use of nanoparticles as therapeutic agents is gaining importance because of their ability to attack bacterial membranes without evoking resistance. We have synthesized the catanionic surfactants Coum12–Coum18, based on fluorescent lactose-derivative organic salts using low-cost starting materials. In water, they self-assemble spontaneously to form stable aggregates at a physiological pH. The antibacterial properties of Coum12–Coum18 were investigated towards multi-drug-resistant Gram-positive bacteria (Staphylococcus aureus and Enterococcus faecalis) and Gram-negative bacteria (Escherichia coli and Pseudomonas aeruginosa). Compound Coum18 was found to have both a bacteriostatic and a bactericidal activity towards Gram-positive bacteria, although the values of both the MIC and MBC (32 μg mL−1) suggest only a topical use of the molecule. The valuable results that were found provide a challenging task for further investigation aimed at the development of this class of antibacterial drugs. In vitro fluorescence microscopy gave insight into the interaction between the aggregates and the cellular membranes on HeLa and CHO cells.

To the Editor: Streptococcus suis, a major swine pathogen worldwide, is emerging as a zoonotic agent capable of causing a variety of serious infections in swine as well as in persons exposed to pigs or to pork products. These infections include meningitis, septicemia, pneumonia, endocarditis, arthritis, and septic shock (1,2). Despite recent outbreaks among persons in China, S. suis disease in humans is a rare, probably underdiagnosed infection that usually occurs as sporadic cases (1,2). Persons in close occupational or accidental contact with pigs or pork products and those who eat uncooked or undercooked pork may be at higher risk than others. However, most infected persons are likely healthy carriers, and S. suis is believed to induce overt disease (especially meningitis) in only some circumstances (2). We describe a case of S. suis meningitis in a 68-year-old man from Sardinia, Italy, who had no reported contact with swine, other animals, or any animal products; the patient also had cancer, which was discovered incidentally during the workup. In November 2007, the patient was hospitalized with a 48-hour history of fever, headache, nausea, and general malaise. Physical examination showed impaired consciousness, nuchal rigidity, and a temperature of 39.5°C. Laboratory findings were 20,700 leukocytes/mm3 with 92% neutrophils, glucose 95 mg/dL, and C-reactive protein 375 mg/L. Cerebrospinal fluid (CSF) analysis demonstrated 240 leukocytes/μL with 80% polymorphonuclear cells, glucose 24 mg/dL, and protein 277 mg/dL. A computed tomography scan of the head showed no abnormal findings. Gram stain of CSF showed gram-positive cocci, mostly in pairs (Figure). Figure Gram-positive cocci, mostly in pairs, in cerebrospinal fluid from a 68-year-old man with Streptococcus suis meningitis. Magnification ×1,000. Empirical therapy consisted of intravenous ceftriaxone (2 g twice a day) and oral chloramphenicol (2 g once a day). On day 5, α-hemolytic streptococci were isolated from CSF on sheep blood agar and identified as S. suis by using APIStrep (bioMerieux, Marcy l’Etoile, France). Serotyping, performed by slide agglutination with specific antiserum (Statens Serum Institute, Copenhagen, Denmark), identified the isolate as serotype 2. Antimicrobial drug­­–susceptibility testing, performed according to guidelines of the Clinical and Laboratory Standards Institute (www.clsi.org), indicated susceptibility to penicillin, ceftriaxone, chloramphenicol, levofloxacin, and vancomycin and resistance to erythromycin (MIC >128 mg/L) and tetracycline (MIC 16 mg/L). Erythromycin resistance was constitutive and was mediated by the erm(B) determinant; tetracycline resistance was mediated by tet(W). Multilocus sequence typing (http://ssuis.mlst.net) assigned the S. suis isolate to sequence type (ST) 1. The patient, a retired welder, denied any recent occupational or even occasional contact with swine or other animals and had no history of eating raw or undercooked pork. The patient’s condition improved; chloramphenicol was discontinued on day 10, but the 14-day course of ceftriaxone was completed. On day 6, the patient became afebrile but had dizziness and deafness; a formal audiology evaluation on day 9 showed severe bilateral sensorineural high-frequency hearing loss (–80 dB) that improved after a short course of dexamethasone. However, the patient was not discharged because of the lung mass found on initial chest radiograph. Computed tomography scan, bronchoscopy, and histopathologic findings led to diagnosis of the mass as an advanced-stage squamous cell carcinoma. The meningitis had common and uncommon features. The common features were hearing loss, a typical outcome of S. suis meningitis independent of early antimicrobial drug administration (1,2); serotype 2, the most frequent and virulent serotype in swine and in humans (1,2); ST1, belonging to the ST1 complex, strongly associated with S. suis meningitis isolates (2,3); and erm(B)-mediated erythromycin resistance, widespread in this species (4). The uncommon features were tetracycline resistance mediated by tet(W), increasingly detected in gram-positive and in gram-negative bacteria (5) but never previously reported in S. suis or in other major streptococcal pathogens, where common determinants are tet(M) and tet(O); and lack of evidence for recent contact with swine, other animals, or swine (pork) products. Two previous cases of human S. suis meningitis in Italy (6,7) and other recent cases from Europe (8,9) were related to occupational exposure. However, the patient reported here also had cancer, and malignancy has been indicated as a predisposing factor for the development of severe S. suis disease in humans (2). These findings appear to be consistent with the recent suggestion of new epidemiologic patterns of infection caused by this organism (2). S. suis may become an opportunistic pathogen in persons who are under stress or who have immunodeficiency, and it has been increasingly isolated from mammalian species other than pigs, from birds, and from the environment. As also discussed in a recent survey (10), the possibility cannot be excluded that a patient with S. suis infection may be unaware or have no memory of previous exposure to animals. Alternatively, because asymptomatic carriage of S. suis has been documented in humans (2) and is believed to contribute to its transmission (10), the possibility should also be considered that the infection may be a reactivation, possibly favored by malignancy, of latently colonizing S. suis.

The Mycobacterium fortuitum group of rapidly growing nontuberculous mycobacteria is an uncommon cause of renal infection, particularly in otherwise healthy hosts. We describe a case of nephritis due to M. fortuitum in an immunocompetent woman with a clinical and radiological diagnosis of renal tuberculosis.

The synthesis and biological activity of selenophenfurin (5-beta-D-ribofuranosylselenophene-3-carboxamide, 1), the selenophene analogue of selenazofurin, are described. Glycosylation of ethyl selenophene-3-carboxylate (6) under stannic chloride-catalyzed conditions gave 2- and 5-glycosylated regioisomers, as a mixture of alpha- and beta-anomers, and the beta-2,5-diglycosylated derivative. Deprotected ethyl 5-beta-D-ribofuranosylselenophene-3-carboxylate (12 beta) was converted into selenophenfurin by ammonolysis. The structure of 12 beta was determined by 1H- and 13C-NMR, crystallographic, and computational studies. Selenophenfurin proved to be antiproliferative against a number of leukemia, lymphoma, and solid tumor cell lines at concentrations similar to those of selenazofurin but was more potent than the thiophene and thiazole analogues thiophenfurin and tiazofurin. Incubation of K562 cells with selenophenfurin resulted in inhibition of IMP dehydrogenase (IMPDH) (76%) and an increase in IMP pools (14.5-fold) with a concurrent decrease in GTP levels (58%). The results obtained confirm the hypothesis that the presence of heteroatoms such as S or Se in the heterocycle in position 2 with respect to the glycosidic bond is essential for both cytotoxicity and IMP dehydrogenase inhibitory activity in this type of C-nucleosides.

Among a series of unmodified phosphodiester (PO)-oligodeoxynucleotides (PO-ODNs) complementary to some of the human immunodeficiency virus type 1 (HIV-1) regulatory genes, several PO-ODN sequences complementary to the vpr gene (PO-ODNs-a-vpr, where a-vpr is the antisense vpr sequence) emerged as potent inhibitors (at concentrations of 0.8 to 3.3 microM) of HIV-1 multiplication in de novo infected MT-4 cells, while they showed no cytotoxicity for uninfected cells at concentrations up to 100 microM. Unlike phosphorothioate counterparts, PO-ODN-a-vpr sequences were not inhibitory to HIV-2 multiplication in de novo infected C8166 cells and neither prevented the fusion between chronically infected and bystander CD4+ cells nor inhibited the activity of the HIV-1 reverse transcriptase in enzyme assays. Moreover, they were not inhibitory to HIV-1 multiplication in chronically infected cells. Delayed addition experiments showed that PO-ODNs-a-vpr inhibit an event in the HIV-1 replication cycle following adsorption to the host cell, but preceding reverse transcription. Structure-activity relationship studies indicated that the antiviral activity of the test PO-ODN-a-vpr sequences is not related to an antisense mechanism but to the presence, within the active sequences, of contiguous guanine residues. Physical characterization of the test PO-ODNs suggested that the active structure is a tetramer stabilized by G quartets (i.e., four G residues connected by eight hydrogen bonds).

The study evaluated the performances of a commercial multiplex PCR assay, the Seegene Seeplex STI Master Panel 3, for Candida spp. identification. Eighty clinical strains of Candida spp. were identified with this system and a homemade multiplex PCR assay. The results were also compared with those obtained with two phenotypic methods. The study provided a preliminary evaluation of a multiplex assay from Seegene that uses capillary electrophoresis as the detection of amplified products. The Seeplex assay was found to be a rapid and useful method for identifying large numbers of yeast isolates in the clinical laboratory context.

Earlier investigations on laboratory and pilot plant scale have shown the resort to microorganisms to be a practicable approach to the problem of purifying mineral dressing plant reject waters from residual flotation reagents and/or metal ions. In spite of the proven effectiveness of this method, one major drawback, namely the pathogenicity of some microorganisms, has so far hampered its application on a commercial scale. A research programme, aimed at developing a microbial reject water purification technique utilizing non-pathogenic strains was thus drawn up and is currently being implemented. Strains such as Bacillus subtilis, Saccharomyces cerevisiiae, Bifidobacterium bifidum, Lactobacillus acidophilus and Acinetobacter calcoaceticus , which are not harmful to human health, some of them being commonly found in the human intestine, have been successfully tested for removing alkylsulphates, alkylamines and fany acids from solutions simulating flotation plant tailings waters. Removals as high as 90% in less than 48 h can be easily achieved with no nutrient requirements, since in most cases the flotation reagent residue to be removed is metabolized by the microorganisms themselves.

21184 Background: The prognostic value of flow cytometry DNA ploidy and Ki-67 expression in colorectal carcinomas has not been defined yet. The study tries to correlate tumoral DNA ploidy and proliferative activity (Ki-67) with therapy response, Overall Survival (OS), Disesase Specific Survival (DSS) and Disease Free Survival (DFS). Methods: From 01/09/02 to 30/06/05, 3 samples of cancer tissue and 1 sample of normal mucosa has been collected from all operating pieces of colorectal cancer. These samples were frosen and later disgregated, treated and coloured with Propidium Iodide. DNA ploidy was evaluated by FACSCalibur cytofluorimetry. Normal mucosa tissue was our internal control. Ki-67 was evaluated by immuno-histochemistry in all tumoral samples. All the patients were cured with chemo- and/or radiotherapy in our divisions. Results: 67 patients (M/F 35/32); median age was 70; staging: 19% I, 33% II, 30% III, 18% IV. We found aneuploidy in 65,7% of carcinoma and Ki-67 median expression was 55%. DNA tumoral heterogeneity was present in 27% of patients. DNA aneuploidy correlates with advanced disease stage at diagnosis (p55%) do not show evident disease versus 100% of patients with DNA diploidy and lower Ki-67. Tumoral aneuploidy correlates in a significative way with lower OS (48% vs 89% of diploid patients), lower DSS (tumor death happened just in patients with aneuploid DNA in every disease stages), with lower DFS (18% vs 86% of diploid patients). Univariated analysis stated that aneuploidy determinates an Odd Ratio=5,7 to develop disease progression (p=0,033). At the moment Ki-67 expression with 55% cut-off does not seem to correlate with OS, DSS and DFS. Conclusions: Preliminar results (the study is still in progress) seem to suggest that cytofluorimetric DNA-ploidy has a prognostic and predictive significance in colo-rectal carcinomas. Ki-67 expression (immuno-histochemistry) has an uncertain significance. The small number of patients and the short follow-up do not allow us to reach any definitive conclusions, but the study is worth to go on. No significant financial relationships to disclose.

The globin CACCC boxes are absolutely required for the appropriate regulation of the β-like globin genes. While the β-globin CACCC box binds EKLF/KLF1, a likely adult switching factor, analogous factors, interacting with the γ-globin gene and predicted to regulate the fetal stage of hemoglobin switching, have so far been elusive. By using yeast one hybrid assay, we have isolated four KLFs, KLF1, 2, 4, and 6, that bound the γ-CACCC bait. To establish their role in globin regulation and in the switching of hemoglobins, these factors were compared to four other KLFs already established or putative globin regulators, KLF3, 11, 13 and 16, mainly evaluating their ability to bind and transactivate the ε-, γ- and β-globin gene. γ-CACCC binding at variable intensities was confirmed in band shift assay for all four isolated KLFs, for KLF3 and, faintly, for KLF13. The ε- and β-CACCC were bound by the same factors with similar affinities with the exception of KLF3 and KLF13 that bound stronger to the β- and ε- than to the γ-CACCC box. On the other hand, KLF11 and 16 did not produce any specific complex in band shift assays with anyone of the globin CACCC boxes. More relevant differences were observed among the factors in the transactivation of single and dual luciferase reporters in both K562 and MEL cells. In these assays, most factors presented peculiar modulatory properties and specific promoter tropism. Several factors presented bidirectional activity displaying in the same time the capacity to stimulate and repress different globin promoters. KLF1 and 4 were the strongest stimulators of the β-globin promoter in both cell lines, whereas KLF2 activated the β-promoter only in K562 cells. KLF1 and especially KLF4 consistently repressed ε-globin expression especially in MEL cells. KLF3 behaved always as a general globin repressor in MEL cells, but acted as a weak stimulator of the γ- and ε-promoter in K562 cells. KLF4 was the strongest inhibitor of the ε-globin gene. KLF13 significantly stimulated the γ-promoter in both cell lines, whereas KLF3, 4 and 6 showed statistically significant stimulation only in MEL cells. By RT-PCR analysis we found that KLFs were highly variable in their tissue expression and that KLF1, 3 and 13 had the highest expression in erythroid tissues. Thus the level of tissue expression should ultimately determine which factors are really active in physiological conditions. Taken together our binding and expression studies suggest that several KLFs have the potential to modulate the activity of the globin genes and that the resulting globin expression will depend on the vectorial sum of the relative activities of the factors expressed at any given time of development. Furthermore, as some KLFs, like KLF1 and 4, exert opposite effects on fetal and adult globin genes, their role in hemoglobin switching may be direct and not only dependent on their ability to mediate promoter competition for the LCR.

Helicobacter pylori since Marshall and Warren's discovery has been an object of interest of gastroenterologists and many researchers of other specialties. What needs to be highlighted is also the growing interest of dentists in the role of oral residue of H. pylori in oral pathologies such as burning mouth syndrome, periodontitis and gingivitis. With the development of medical techniques more studies using highly specific diagnostic methods are performed in order to determine the transmission pattern of bacterial infection. Suggested faecal-oral and oral-oral routes of bacterial transmission raised interest in molecular biology methods as tools for the study of these environments. Additionally, co-existence of helical and coccoidal forms of H. pylori in the mentioned niches raised the question whether the latter is potentially pathogenic. This is why molecular biology is now giving a great opportunity to explore parts of the human body that could not have been diagnosed before using only gold standard diagnostic methods. Molecular techniques have shown their usefulness in examining the potential virulence of coccoid forms of bacterium. This review was created also to summarize the knowledge about molecular methods, especially different PCR techniques, as diagnostic tools that can help medical teams during regular diagnosis of gastritis.