Bloody Diarrhea and Shiga Toxin-Producing Escherichia coli Hemolytic Uremic Syndrome in Children: Data from the ItalKid-HUS Network.

OBJECTIVE: To analyze the results of an enhanced laboratory-surveillance protocol for bloody diarrhea aimed at identifying children with Shiga toxin-producing Escherichia coli (STEC) infection early in the course of the disease toward the early identification and management of patients with hemolytic uremic syndrome (HUS). STUDY DESIGN: The study (2010-2019) involved a referral population of 2.3 million children. Stool samples of patients with bloody diarrhea were screened for Shiga toxin (Stx) genes. Positive patients were rehydrated and monitored for hemoglobinuria until diarrhea resolved or STEC-HUS was diagnosed. RESULTS: A total of 4767 children were screened; 214 (4.5%) were positive for either Stx1 (29.0%) or Stx2 (45.3%) or both Stx1+2 (25.7%); 34 patients (15.9%) developed STEC-HUS (0.71% of bloody diarrheas). Hemoglobinuria was present in all patients with HUS. Patients with Stx2 alone showed a greater risk of STEC-HUS (23.7% vs 12.7%) and none of the patients with Stx1 alone developed HUS. During the same period of time, 95 other patients were diagnosed STEC-HUS but were not captured by the screening program (26 had nonbloody diarrhea, 11 came from areas not covered by the screening program, and 58 had not been referred to the screening program, although they did meet the inclusion criteria). At HUS presentation, serum creatinine of patients identified by screening was significantly lower compared with that of the remaining patients (median 0.9 vs 1.51 mg/dL). CONCLUSIONS: Nearly 1% of children with bloody diarrhea developed STEC-HUS, and its diagnosis was anticipated by the screening program for Stx. The screening of bloody diarrhea for Stx is recommended, and monitoring patients carrying Stx2 with urine dipstick for hemoglobinuria is suggested to identify the renal complication as early as possible.

Family Clusters of Shiga Toxin-producing Escherichia coli Infection: An Overlooked Source of Transmission. Data From the ItalKid-Hus Network.

BACKGROUND: The aim of the present work was to investigate family clusters of Shiga toxin-producing Escherichia coli (STEC) infection among the household members of STEC positive patients, identified within a screening program of bloody diarrhea (BD) for STEC in Northern Italy. METHODS: Stool samples from patients with BD or BD-associated-hemolytic uremic syndrome (HUS) and related households were investigated by molecular and bacteriologic methods to detect and characterize the virulence profile of STEC and Pulsed Field Gel Electrophoresis analysis were done on isolates. RESULTS: Thirty-nine cases of STEC infection (isolated BD in 16, BD-associated-HUS in 23) were considered, and a total of 130 stool samples from 1 to 8 households of the index patient were analyzed. The prevalence of positivity was higher in siblings (34.8%, 8/23) than in mothers (20%, 7/35), grandparents (9.5%, 2/21), fathers (8.8%, 3/34) or other households. In 14 clusters (36%), one or more household shared a STEC with the same virulence profile (stx, eae, serogroup) as the index case. In 7 clusters, STEC strains isolated from at least 2 subjects also shared identical Pulsed Field Gel Electrophoresis profile. The frequency of household infection does not appear to be associated to the index case's illness (HUS or BD), nor with the serotype or with the virulence profile of the involved STEC (stx2 or stx1-stx2). CONCLUSIONS: Our study shows that STEC infections, most likely related to human-to-human transmission, are common among households of patients with STEC BD or HUS and underlines the importance of extending the epidemiologic investigations to all family members, as the index case may not always be the primary infection in the family.

Drug Loaded Gingival Mesenchymal Stromal Cells (GinPa-MSCs) Inhibit In Vitro Proliferation of Oral Squamous Cell Carcinoma.

Human mesenchymal stromal cells (MSCs) have been widely investigated both for regenerative medicine and their antinflammatory/immunomodulatory capacity. However, their ability to home pathological tissues suggested the development of strategies for using MSCs as carrier to deliver drug into tumor microenvironment. MSCs obtained from different tissues can be loaded in vitro with anti-cancer drugs by a simple procedures. In this report, we studied MSCs isolated and expanded from gingival papilla (GinPa-MSCs), by testing their ability to uptake and release three important anti-neoplastic drugs: Paclitaxel (PTX), Doxorubicin (DXR) and Gemcitabine (GCB). The efficacy of drugs releasing GinPa-MSCs was studied on a pancreatic cancer cell line and confirmed in vitro against a line of tongue squamous cell carcinoma (SCC154). Our results demonstrated that GinPa-MSCs efficiently incorporate the drugs and then released them in active form and in sufficient amount to produce a dramatic inhibition of squamous cell carcinoma growth in vitro. If compared with other MSCs sources, the collection of GinPa-MSCs is poorly invasive and cells can be easily expanded and efficiently loaded with anti cancer drugs. In particular, gemcitabine loaded GinPa-MSCs provide a good "cell-mediated drug delivery system" for a future potential application in the context of the oral oncology.

Cytotoxic and Antimicrobial Activities of Cantharellus cibarius Fr. (Cantarellaceae).

Antibacterial and cytotoxic activities of cyclohexane, dichloromethane, methanol, and aqueous extracts of Cantharellus cibarius were tested. Broth microdilution assay was performed against 10 bacterial strains (Staphylococcus aureus, S. epidermidis, Micrococcus luteus, Bacillus subtilis, Enterococcus feacalis, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella abony), with emphasis on Helicobacter pylori. Methanol extract was the most active against H. pylori strains with minimal inhibitory concentration values between 4 and 32 µg/mL. All extracts were active against antibiotic resistant H. pylori. Methanol and aqueous extracts had no cytotoxicity against tested cell lines, whereas cyclohexane and dichloromethane extracts were active against HeLa and N87 cells, but also against healthy MRC-5 cells (IC50 39.26 ± 1.24-134.79 ± 0.01 µg/mL). The tested aqueous extracts have shown 68% of angiotensin-converting enzyme inhibitory activity in doses of 1.25 mg/mL. Chemical analysis has shown the presence of linoleic, cis-vaccenic, and oleic acids, sterols, ß-glucans, and polyphenolic compounds.

Paclitaxel-releasing mesenchymal stromal cells inhibit in vitro proliferation of human mesothelioma cells.

INTRODUCTION: Malignant pleural mesothelioma (MPM) is a rare fatal asbestos-related malignancy originating in the mesothelial cells of the pleura. A platinum-based doublet containing a third-generation antifolate is the front-line standard of care whilst there are no approved second-line treatments for MPM which remains a disease setting to test the efficacy of new therapeutic agents. METHODS: Bone marrow mesenchymal stromal cells (BM-MSCs) were loaded with pemetrexed (PMX) and paclitaxel (PTX) according to a standardized procedure. Drug release by both PMX- and PTX-primed BM-MSCs (BM-MSCs/PMX and BM-MSCs/PTX) was tested on the in vitro proliferation of a panel of tumor cell lines including NCI-H28 mesothelioma. RESULTS: The in vitro anticancer activity of pure PTX was significantly higher than that of PMX against all the cell lines tested (14.7 times higher than that of PMX against NCI-H28). Whereas BM-MSCs did not take up and release PMX in amounts effective on mesothelioma, PTX-loaded BM-MSCs dramatically inhibited mesothelioma proliferation. CONCLUSIONS: PTX-primed mesenchymal stromal cells successfully inhibit the in vitro proliferation of human mesothelioma cells. Further studies and in vivo testing are required to confirm our preliminary in vitro results as a potential new mesothelioma therapy based on cell drug delivery.

In vitro activity of artemisone and artemisinin derivatives against extracellular and intracellular Helicobacter pylori.

The in vitro activity of the new artemisinin derivative artemisone as well as other molecules of the same class against Helicobacter pylori and their effects when combined with standard antibiotics were evaluated. Since H. pylori can be internalised into gastric epithelial cells, the effects of artemisinin, dihydroartemisinin and artemisone against intracellular H. pylori were also investigated. Bacteriostatic [minimum inhibitory concentration (MIC)] and bactericidal [minimum bactericidal concentration (MBC)] activities were assessed against 24 clinical strains of H. pylori with different antibiotics susceptibilities. Artemisone showed MIC50 and MIC90 values of 0.25 mg/L and 0.5 mg/L, respectively, and an MBC50 value of 0.5 mg/L. Artemisone was synergistic with amoxicillin in 60% of strains, with clarithromycin in 40% and with metronidazole in 20%. There was no interaction between artemisone and omeprazole or bismuth citrate. Against intracellular H. pylori, only dihydroartemisinin at 2× MIC caused a 1 log10 CFU decrease after 18 h and 24 h of incubation. This is the first demonstration in vitro of the activity of artemisinin derivatives against intracellular H. pylori and indicates that artemisone has the potential to be efficacious for the treatment of H. pylori infection, especially in combination with antibiotics.

Anti-Helicobacter pylori Activity of Four Alchemilla Species (Rosaceae).

The present study evaluated the anti-Helicobacterpylori activity of Alchemilla glabra Neygenf. (A. sect. Alchemilla), A. monticola Opiz (A. sect. Plicatae S.E. Fröhner), A. fissa Günther & Schummel (A. sect. Calycinae (Buser) Buser) and A. viridiflora Rothm. (A. sect. Calycinae), and identified ellagic acid and quercetin-3-O-ß-glucoside. Anti-H. pylori activity was tested against ten clinical isolates and one reference strain (ATCC 43504). The methanol extracts were more active than the dichloromethane and cyclohexane extracts. The ranges of concentrations were between 4 µg/mL for methanol extracts of A. viridiflora, A. glabra and A. monticola, and 256 µg/mL for cyclohexane extracts of A. viridiflora, A. glabra and A. fissa. The best overall activity was obtained with A. monticola extracts. No significant difference was found in the ellagic acid contents of the methanol extracts of the tested Alchemilla species (0.2-0.3 mg/mL), and anti-H. pylori activity was similar (4-32 µg/mL). Ellagic acid exhibited strong activity at very low concentrations (0.125-0.5 µg/mL), while the second identified compound, quercetin-3-O-ß-D-glucoside, was also very active in concentration of 2-16 µg/mL.

Bioactive compounds of Crocus sativus L. and their semi-synthetic derivatives as promising anti-Helicobacter pylori, anti-malarial and anti-leishmanial agents.

Crocus sativus L. is known in herbal medicine for the various pharmacological effects of its components, but no data are found in literature about its biological properties toward Helicobacter pylori, Plasmodium spp. and Leishmania spp. In this work, the potential anti-bacterial and anti-parasitic effects of crocin and safranal, two important bioactive components in C. sativus, were explored, and also some semi-synthetic derivatives of safranal were tested in order to establish which modifications in the chemical structure could improve the biological activity. According to our promising results, we virtually screened our compounds by means of molecular modeling studies against the main H. pylori enzymes in order to unravel their putative mechanism of action.