Ceftazidime/avibactam-resistant meropenem-susceptible KPC-producing Klebsiella pneumoniae: Analysis of cases and evaluation of in vitro activity of fosfomycin-containing combinations.

OBJECTIVES: Little is known regarding outcomes and optimal therapeutic regimens of infections caused by Klebsiella pneumoniae carbapenemase-producing Klebsiella pneumoniae (KPC-Kp) resistant to ceftazidime/avibactam (CZA) and susceptible to meropenem (MEM). Although susceptible to MEM in vitro, the possibility of developing MEM resistance overtime is a concern. We describe the clinical characteristics of patients with colonization/infection due to KPC variants with a focus on the in vitro activity of fosfomycin (FOS)-containing combinations. METHODS: Patients with colonization/infection due to a KPC variant were included. Fosfomycin susceptibility was performed by agar dilution method. Synergistic activity of FOS-based combinations was evaluated by gradient strip-agar diffusion method. The emergence of in vitro MEM resistance was also tested. RESULTS: Eleven patients were included: eight with infection [four with ventilator-associated pneumonia and four with bloodstream infections] and three with colonization. Previous therapy with CZA was administered to all patients (with a mean cumulative duration of 23 days). All subjects with infection received meropenem, in monotherapy (n = 4) or with amikacin (n = 2) or fosfomycin (n = 2), and achieved clinical cure. A new CZA-susceptible and MEM-resistant KPC-Kp strain was subsequently isolated in three patients (27.3%). Meropenem/vaborbactam (MVB) showed high in vitro activity, while FOS+MEM combination was synergistic in 40% of cases. In vitro resistance to MEM was observed with maintenance of CZA resistance. CONCLUSIONS: Detection of KPC variants may occur within the same patient, especially if CZA has been previously administered. Although clinical success has been obtained with carbapenems, the emergence of MEM resistance is a concern. Fosfomycin plus meropenem is synergistic and may be a valuable combination option for KPC variants, while MVB may be considered in monotherapy. The detection of KPC variants in an endemic setting for KPC-Kp represents a worryingly emerging condition. The optimal therapeutic approach is still unknown and the development of meropenem resistance is of concern, which may lead to therapeutic failure in clinical practice. In these cases, the addition of fosfomycin to meropenem, or a more potent antibiotic, such as meropenem/vaborbactam, may be valuable therapeutic options.

Animal welfare official inspections: farmers and inspectors shared concerns.

Official inspections to check the compliance of farms with European legislation to protect farm animals are often perceived negatively by farmers. In addition, the inspections have a limited effect on improving farm compliance. We looked at the perceptions of both farmers and their inspectors about animal welfare and the inspections in a case study of dairy production in France. The identification of gaps and commonalities between both parties should help us to propose improvements in the inspection method by which inspections could more likely encourage compliance with animal welfare legislation. To achieve this aim, we conducted semi-structured interviews with 22 dairy farmers and their 19 inspectors. Both farmers and inspectors described animal welfare in terms of the state of the animal and of the living conditions and care provided to them. The majority of farmers found that the official checklist used by the inspectors is inappropriate to assess the welfare of their animals; inspectors themselves reported that they often use their own criteria and indicators (often based on the observation of animals) in addition to the official checklist. Both groups disagreed with some requirements of the legislation. These findings suggest that the content and background of the legislation to protect animals should be made clearer to both farmers and inspectors and that these two groups of actors should be involved in the definition of key points to be checked on farms, with special attention to animal-based indicators. All this could improve farmers' engagement with the results of the inspections and, hopefully, could lead to better compliance with legislation and improvements in animal welfare on farms.

Successful Treatment of Kaposi Sarcoma-Associated Herpesvirus Inflammatory Cytokine Syndrome After Kidney-Liver Transplant: Correlations With the Human Herpesvirus 8 miRNome and Specific T Cell Response.

After transplant, patient infection with human herpesvirus 8 (HHV-8) and Kaposi sarcoma-associated herpesvirus (KSHV) is known to cause aggressive tumors and severe nonneoplastic complications. These latter syndromes are driven by HHV-8/KSHV lytic reactivations and related hyperinflammatory host responses typically characterized by high viral loads, elevated levels of cytokines and other inflammation biomarkers, cytopenia, organ failure, high fever, and worsening conditions (with no evidence of B cell neoplasias). These disorders are associated with a high mortality rate, often due to lack of prompt diagnosis, effective therapeutic approaches, and adequate follow-up. These features resemble most of those defining the so-called KSHV-associated inflammatory cytokine syndrome (KICS), which was recently recognized in patients positive for human immunodeficiency virus (HIV). In this report, we describe-for the first time-a case of a KICS-like nonneoplastic recurrent complication occurring after transplant in an HIV-negative patient that was successfully treated by a combination of anti-CD20 monoclonal therapy, antivirals, and modification of the immunosuppressive regimen. In addition to clinical and laboratory findings collected during 3-year follow-up, we report novel experimental data on HHV-8-specific T cell dynamics and circulating microRNA profile, showing correlations with clinical course and other laboratory markers (including viral load, C-reactive protein, and cytokine levels), providing useful information about abnormal cellular and cytokine dynamics underlying HHV-8-associated inflammatory disorders in posttransplant patients.

Efficacy of safety catheter devices in the prevention of occupational needlestick injuries: applied research in the Liguria Region (Italy).

Healthcare workers who use or may be exposed to needles are at risk of needlestick injuries, which can lead to serious infections by bloodborne pathogens. These injuries can be avoided by eliminating the unnecessary use of needles and using safety devices. The present study was aimed at evaluating the impact of a safety-engineered device, with passive fully automatic needlestick protection, on the rate of needlestick injuries among healthcare workers. The setting of the study was a network of five public healthcare institutions situated in a Northern Italian Region. Data on the type of device, the number of employees and the number of catheter devices used per year were collected through regular meetings with healthcare workers over a period of five years. The most notable result of this study was the huge risk reduction associated with safety devices. Indeed, the risk of needlestick injuries due to conventional devices was found to be 25-fold higher than that observed for safety devices. However, it is noteworthy that a considerable part of this excess can be explained by the different background number of devices used. Moreover, descriptive analysis suggested that individuals with a poor/moderate training level had a lower risk than those with good/high training, though the difference was not statistically significant. In conclusion, there is convincing evidence of a causal connection between the introduction of safety devices and the reduction in needlestick injuries. This consideration should prompt the introduction of safety devices into daily clinical practice.

Structural comparison of the interaction of tubulin with various ligands affecting microtubule dynamics.

Microtubules (MTs), which are highly dynamic assemblies of the protein tubulin, play important and diverse roles in eukaryotic cells. MT dynamics are regulated during the cell cycle by interacting with a large number of endogenous cellular regulators. In addition, many anti-tumour drugs and natural ligands that interact directly with tubulin are able to either stabilise or destabilise MTs and to disrupt the normal dynamics. Herein, we compare the structures of tubulin when complexed with different ligands in order to analyse: (i) various binding-sites of the protein and different positions of ligands within the microtubule (ii) the diverse effect on the microtubule dynamics. The structures and data given are essential for understanding tubulin-ligand interactions and their influence on the regulation of the microtubule system.

Values and public acceptability dimensions of sustainable egg production.

The attributes of egg production that elicit values-based responses include the price and availability of eggs, environmental impacts, food safety or health concerns, and animal welfare. Different social groups have distinct interests regarding the sustainability of egg production that reflect these diverse values. Current scientifically based knowledge about how values and attitudes in these groups can be characterized is uneven and must be derived from studies conducted at varying times and using incomplete study methods. In general, some producer and consumer interests are translated through markets and are mediated by market mechanisms, whereas others are poorly reflected by economic behavior. An array of survey and focus group research has been performed to elicit consumer and activist beliefs about performance goals they would expect from an egg production system. These studies provide evidence that consumers' market behavior may be at odds with their ethical and political beliefs about performance goals.

CDMS (Clinical Data Mining Software): a cytokinome data mining system for a predictive medicine of chronic inflammatory diseases.

The cytokines, main players of the chronic inflammation progression leading to serious diseases such as diabetes or cancer, represent a target for better clinical prognosis and innovative therapeutic strategies. To investigate the immunopathogenetic progression of these diseases, the evaluation of serum cytokines profiles made of many different proteins is much more informative than single protein measurements. We developed a Clinical Data Mining Software to collect cytokine profiles evaluated on healthy subjects and patients by multiplex immunoassays also annotated with their clinical and laboratory data, to compare patient profiles by statistical tools and to evaluate their disease progression.

Advanced swine manure treatment and utilization in Brazil.

Animal production has changed from subsistence to an industrial model, lowering production costs but giving rise to higher potential environmental impact. When the effluents are not correctly managed, serious pollution events can occur. In Brazil liquid manure is commonly stored in reception pits or covered lagoons (biodigestors), followed by land application as a biofertilizer. In some regions there is an excess of manure due to low soil support capacities, and in these cases new technologies have to be adopted to export or treat the excess effluent. Manure storage time in pits/covered lagoons and new polymers to separate the solid fraction have been studied in Brazil. Treatment technologies, like swine manure treatment systems (SMTS), have been developed from a technical and economical point of view to optimize the processes and give a technological alternative to pork producers increasing production while reducing environmental impact.

Ligand binding and cytochemical analysis of estrogen and progesterone receptors in relation to follow-up in patients with breast cancer.

Soluble and nuclear estrogen receptor (ER) content was measured by ligand binding assay, and estrogen and progesterone receptors by immunohistochemical assays (ER-ICA and PR-ICA) in 214 patients with breast cancer recruited at the "M. Ascoli" Cancer Hospital Centre in Palermo, Sicily, to assess the discriminant and predictive value of these parameters. On follow-up, data from both ER-ICA and PR-ICA showed a statistically significant difference, PR-positive patients having longer disease-free (DSF) and overall (OS) survival than PR-negative ones. Conversely, ER status did not correlate significantly with both DFS (P = 0.6) and OS (P = 0.2). In particular, PR-positive patients had 59 +/- 18 months DFS and 67 +/- 12 months OS, compared to 51 +/- 22 months DFS and 57 +/- 17 months OS of PR-negative cases. The present evidence implies that a PR-negative status identifies breast cancer patients with early relapse, as also suggested by previous studies. It also agrees with the results of ligand binding assay of ER, where ER status is a good discriminant and predictor of response to endocrine treatment, but is unable to anticipate early relapse in breast cancer patients. Evidence that PR status is a statistically significant prognostic indicator deserves further study to ascertain whether or not PR should be regarded as an ER-dependent parameter or be related to other biological variables such as growth factor (e.g., EGF), oncogene (e.g., Her2/Neu), or tumor suppressor gene (e.g., p53) products.

The Mediet Project.

Preliminary evidence from a case control study of healthy postmenopausal women living in Palermo, Sicily, is presented to investigate the potential impact of a traditional Mediterranean diet on the risk of developing breast cancer. Of the 230 women who fulfilled specific eligibility criteria, 115 were enrolled in the study based on serum testosterone values equal to or greater than the median population value (0.14 microg/ml). Women were then individually randomized into a diet intervention (n = 58) and a control (n = 55) group. Women in the intervention group attended a weekly "cooking course" for 1 year, being trained by professional chefs in the correct use of the natural ingredients of the traditional Mediterranean diet, including whole cereals, legumes, seeds, fish, cruciferous vegetables, and many others. The intervention group was subsequently instructed to follow the learned diet at home, while the control group was only advised to increase the consumption of fruits and vegetables, as recommended by WHO. The following measures were taken at the beginning, middle, and end of the study: (a) fasting blood and 12-hour urine samples to assay defined hormonal endpoints; (b) height, weight, and circumference of the waist and hip; and (c) a food frequency and computerized 24-hour dietary recall questionnaire. After 1 year, both the control and the intervention groups showed satisfactory compliance rates (81 and 85%, respectively). In addition, preliminary results so far obtained reveal an unequivocal trend towards weight loss, a strong reduction in cholesterol levels, and a psychophysical feeling of well-being by women adopting the Mediterranean diet. The study is currently ongoing to verify the association of changes in serum and urine hormone levels and breast cancer risk in the intervention group.

Phorbol diester 12-O-tetradecanoylphorbol 13-acetate (TPA) up-regulates the expression of estrogen receptors in human THP-1 leukemia cells.

In the present work, we have inspected expression of estrogen receptors (ER) and their regulation by the phorbol diester 12-O-tetradecanoylphorbol 13-acetate (TPA) in a leukemic cell line, the THP-1 cells, using multiple experimental approaches. Firstly, ligand binding assay (LBA) revealed that control (unstimulated) THP-1 cells express type I (high affinity, limited capacity) ER in the nuclear fraction only, whilst treatment of cells with TPA resulted in the appearance of type I ER in the soluble fraction as well, with the 50 ng/ml dose and the 48 h incubation time being the most effective experimental condition. A concomitant increase of type II ER was also seen in both soluble and nuclear cell fractions. Unstimulated THP-1 cells were found to be ER negative by immunocytochemistry; conversely, cells exposed to 50 ng/ml TPA for 48 h stained positively for ER, with the majority of cells having a strong nuclear staining. Scrutiny of ER mRNA expression using reverse transcriptase-polymerase chain reaction showed the presence of a wild type ER transcript in both control and TPA-treated THP-1 cells, though levels of ER mRNA were found to be comparatively higher in the latter. This combined evidence would imply that the TPA-induced differentiation of THP-1 cells is accompanied by the rise of high affinity (type I) ER, suggesting that estrogens may play a role in the regulation of macrophage activity during the inflammatory and/or the immune response.

A study on the role of nitric oxide and iron in 3-morpholino-sydnonimine-induced increases in dopamine release in the striatum of freely moving rats.

1. We showed previously that interaction between NO and iron (II), both released following the decomposition of sodium nitroprusside (SNP), accounted for the late SNP-induced dopamine (DA) increase in dialysates from the striatum of freely moving rats; in addition, we showed that co-infusion of iron (II) with the NO-donor S-nitroso-N-acetylpenicillamine mimicked SNP effects on striatal DA release. 2. In the present study, intrastriatal co-infusion of iron (II) (given as FeSO(4), 1 mM for 40 min) with the NO-donor and potential peroxynitrite generator 3-morpholinosydnonimine (SIN-1) (0.2, 0.5, 1.0 or 5.0 mM for 180 min), potentiated the SIN-1-induced increase in DA concentration in dialysates from the striatum of freely moving rats. Neither alone nor associated with iron (II) did SIN-1 induce changes in dialysate ascorbic acid or uric acid concentrations. 3. Neither co-infusion of a superoxide dismutase mimetic nor uric acid affected SIN-1-induced increases in dialysate DA concentration. 4. Infusion of the iron chelator deferoxamine (0.2 mM for 180 min) decreased dialysate DA and attenuated SIN-1-induced increases in dialysate DA concentrations. 5. These results suggest that iron plays a key role in SIN-1-induced release of striatal DA and do not support any role for either peroxynitrite or superoxide anion in SIN-1-induced release of striatal DA.

Analysis of S-nitroso-N-acetylpenicillamine effects on dopamine release in the striatum of freely moving rats: role of endogenous ascorbic acid and oxidative stress.

1. We showed previously that interaction between NO and iron(II), both released following decomposition of sodium nitroprusside (SNP), accounted for the late SNP-induced dopamine (DA) increase in dialysates from the striatum of freely moving rats. 2. In this study, intrastriatal infusion of the NO-donor S-nitroso-N-acetylpenicillamine (SNAP) (0.2 mM for 180 min) induced a moderate increase in dialysate DA and decreases in ascorbic acid dialysate concentrations; in contrast, SNAP 1 mM infusion induced a long-lasting decrease in both DA and ascorbic acid dialysate concentrations. 3-Methoxy-tyramine (3-MT), dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA), and uric acid levels were unaffected. 3. Co-infusion of ferrous sulphate [iron(II), 1 mM for 40 min] with SNAP either 1 or 0.2 mM (for 180 min), produced a significant increase in both DA and 3-MT dialysate concentrations, but it did not affect decreases in dialysate ascorbic acid levels. All other dialysate neurochemicals were unaffected. 4. Co-infusion of ascorbic acid (0.1 mM) with SNAP (1 mM) for 180 min did not modify SNAP-induced decreases in dialysate DA levels. In contrast, co-infusion of uric acid (1 mM) reversed SNAP-induced decreases in dialysate DA; co-infusion of a superoxide dismutase mimetic delayed SNAP-induced DA decreases for a short period, while co-infusion of the antioxidant N-acetylcysteine (NAC, 0.1 mM) significantly increased dialysate DA. 5. The results of this study show that SNAP induces concentration-related changes in DA dialysate levels. At higher concentrations, SNAP induces non-enzymatic DA oxidation, which is inhibited by uric acid and NAC; ascorbic acid failed to protect dialysate DA from oxidation, probably owing to its promoting effect on SNAP decomposition; exogenous iron(II) may react with NO generated from SNAP decomposition, with a consequent increase in dialysate DA and 3-MT, therefore mimicking SNP effects on striatal DA release.

Methyleugenol in Ocimum basilicum L. Cv. genovese gigante.

Ocimum basilicum cv. Genovese Gigante is the basil cultivar used the most in the production of a typical Italian sauce called pesto. The aromatic composition of plants at different growth stages was determined. Plants from different areas of northwestern Italy were analyzed at 4 and 6 weeks after sowing and showed methyleugenol and eugenol as the main components. The content of these compounds was correlated with plant height rather than plant age. Particularly, methyleugenol was predominant in plants up to 10 cm in height, whereas eugenol was prevalent in taller plants. These results are important in the evaluation of risk to human health posed by dietary ingestion of methyleugenol contained in pesto.

New insights on disintegrin-receptor interactions: eristostatin and melanoma cells.

Viper venom disintegrins have been used frequently to study the cellular receptors which characterize various types of cells, including platelets, endothelial cells and cancer cells. While the majority of such analyses have pointed to involvement of integrin receptors alphavbeta3, alpha5beta1 or alphaIIbbeta3, this may not always be so. Eristostatin, from Eristocophis macmahoni, is a potent inhibitor of ADP-induced platelet aggregation as well as of human and murine melanoma metastases in mouse model systems. This disintegrin requires an RGDW motif, as well as an intact C-terminus, in order to interact with both platelets and four different types of melanoma cells. Eristostatin causes nonmetastatic SBc12 melanoma cells to show higher susceptibility to specific killing by NK-like TALL-104 cells. While it is known that eristostatin binds to alphaIIbbeta3 on platelets, the receptor with which eristostatin binds to the melanoma cells has not yet been identified.

Percent free PSA as an additional measure in a prostate cancer screen.

OBJECTIVE: To assess the ability of the ratio of free prostate specific antigen to total prostate specific antigen (% fPSA) to aid in selection of subjects who would require follow-up studies. DESIGN: Retrospective, nonrandomized analysis measuring total prostate specific antigen (tPSA) and free prostate specific antigen (fPSA) in serum from men enrolled in a community-based prostate cancer screening offered by the Cancer Outreach Program (COP), Christiana Care Health Systems (CCHS), Wilmington, DE. PARTICIPANTS Informed consent was obtained from 172 of the 231 eligible participants. Complete laboratory and follow-up data, excluding ethnic origin, was gathered from 157 participants; therefore, theoretical participation was 75% (172/231) while realized participation rate was 68% (157/231). Criteria for inclusion in this study included ambulatory men of age 40 and older with serum total PSA (tPSA) level >1.9 ng/mL and/or an enlarged or abnormal prostate by digital rectal examination (DRE). Subjects with a history of prostate cancer or prostatitis were excluded. MAIN OUTCOME MEASURES: Laboratory tests to determine tPSA and fPSA were performed on serum samples obtained from consenting participants. Percent fPSA was calculated. Results of clinical findings with respect to each participant's DRE were recorded as normal, suggestive of a benign condition, such as benign prostate hyperplasia, or suspicious for prostate cancer. When available, transurethral ultrasound (TRUS) results and biopsy results were also noted. Each participant's results were evaluated and given one of the following diagnoses: Normal prostate (N), Benign Prostate Hyperplasia (BPH), or Prostate Cancer (PCa). RESULTS: Prevalence of cancer 3/157 = 0.0191; of BPH = 0.688; of N = 0.293. Median values for tPSA for each of these groups were as follows: N, 2.9 ng/mL; BPH, 3.0 ng/mL; and PCa, 6.3 ng/mL; (p = 0.079). Median values for fPSA were as follows: N, 0.6 ng/ mL; BPH, 0.5 ng/mL; PCa, 0.5 ng/mL; (p = 0.51). Median values for % fPSA were as follows: N, 19%; BPH, 17%; and PCa, 9%; (p = 0.01). Medians were found to differ for % fPSA measurements, but not for tPSA or fPSA values. DRE screening results of 110 subjects were reported as not normal indicating either an enlarged (n = 97) or abnormal prostate gland (n = 13). Nine subjects had normal DRE results with serum tPSA level above 4.0 ng/mL. Using the combination of DRE and tPSA > 4.0 ng/mL as criteria for the recommendation of follow-up studies, 119 of the participants would have been advised to seek additional testing. CONCLUSION: Using these two routine criteria, 119 (119/157; 76%) subjects would be candidates for follow-up procedures, such as transurethral ultrasound (TRUS) and/or sextant prostate biopsy. By adding % fPSA results of < 10% fPSA to tPSA results > 4.0 mg/mL as criteria for follow-up studies, specificity can be improved threefold with sensitivity unchanged. If an abnormal DRE suggestive of malignancy was included as part of the criteria, sensitivity of the diagnostic scheme would reach 100%. Using the triple diagnostic parameters of tPSA > 4.0 ng/mL, the ratio of free prostate specific antigen to total prostate specific antigen <10%, and an abnormal prostate DRE, 16 participants would be recommended for follow-up studies (16/157; 10%). This would eliminate 103 subjects from unnecessary and expensive testing.

Unsymmetrical methylene derivatives of indoles as antiproliferative agents.

Indole-3-carbinol is a natural product which has been shown to reduce the incidence of spontaneous and carcinogen-induced mammary tumours in animals. Eighteen unsymmetrical methylene derivatives of indoles were prepared by reaction of Mannich bases of 7-hydroxycoumarins with substituted indoles in acetic or propionic anhydride. The synthesised molecules were tested in vitro against the MCF7 and MDA-MB-231 breast cancer cell lines by MTT and cell count assays. Results from 16 tested compounds showed that 60% of them exerted some effects against the MDA-MB-231 compared to about 30% towards the MCF7. Among all, the 3-(7'-acetoxy-4-methylcoumarin-8'-yl)methyl-2-methylindole resulted the most effective in both cell lines, compared to indole-3-carbinol. In conclusion, these preliminary results report that some of these compounds might be promising potential antiproliferative agents.

Methyleugenol and eugenol variation in Ocimum basilicum cv. Genovese gigante grown in greenhouse and in vitro.

The variation of the two main aromatic compounds in Ocimum basilicum cv Genovese Gigante grown in greenhouse and "in vitro" was analyzed. The content of methyleugenol and eugenol was correlated to the plants' height rather than to the plants' age and the growth site. Particularly, methyleugenol was prevalent in plants up to 6.5 cm, as plants grew it was replaced by eugenol that was dominant in taller plants. Analysis of basil 20 cm in height showed that methyleugenol is prevalently localized in the low part while eugenol is prevalent in the upper part of the plant. Moreover, a chronic and acute toxicity of methyleugenol was evidentiated in an assay using chicken embryos.