Drosophila phagocytosis – still many unknowns under the surfaceULVILA, JOHANNA; VANHA‐AHO, LEENA‐MAIJA; RÄMET, MIKA
doi: 10.1111/j.1600-0463.2011.02792.xpmid: 21917002
Ulvila J, Vanha‐aho L‐M, Rämet M. Drosophila phagocytosis – still many unknowns under the surface. APMIS 2011; 119: 651–62. In mammals, phagocytosis coordinates host defence on two levels: It acts both as an effector of the innate immunity, as well as an initiator of the adaptive immunity. The fruit fly Drosophila melanogaster (D. melanogaster) lacks the adaptive immune response, and the role of Drosophila plasmatocytes, cells that resemble phagocytosing mammalian macrophages, is limited to innate immune responses. During the past years, several studies have shed light on the role of phagocytosis in the Drosophila host defence. At least in some infection models, the systemic production of potent antimicrobial peptides (AMPs) does not completely compensate for the need for cellular immune responses. As a model, Drosophila offers powerful tools for studying phagocytosis including, large‐scale RNA interference (RNAi) based in vitro screens that can be combined with classical Drosophila genetics. These kinds of approaches have led to important discoveries related especially to microbial recognition by Drosophila plasmatocytes. Events following initial recognition, however, have remained more elusive. This review summarizes the current knowledge on Drosophila phagocytosis focusing on the most recent advancements in the field, and highlighting the benefits the Drosophila system has to offer for research on phagocytosis.
High expression of miR‐21 in tumor stroma correlates with increased cancer cell proliferation in human breast cancerRASK, LENE; BALSLEV, EVA; JØRGENSEN, STINE; ERIKSEN, JENS; FLYGER, HENRIK; MØLLER, SØREN; HØGDALL, ESTRID; LITMAN, THOMAS; SCHNACK NIELSEN, BOYE
doi: 10.1111/j.1600-0463.2011.02782.xpmid: 21917003
Rask L, Balslev E, Jørgensen S, Eriksen J, Flyger H, Møller S, Høgdall E, Litman T, Nielsen BS. High expression of miR‐21 in tumor stroma correlates with increased cancer cell proliferation in human breast cancer. APMIS 2011; 119: 663–73. Low‐risk and high‐risk breast cancer patients are stratified primarily according to their lymph node (LN) status and grading. However, some low‐risk patients relapse, and some high‐risk patients have a favorable clinical outcome, implying a need for better prognostic and predictive tests. Micro RNAs are often aberrantly expressed in cancer and microRNA‐21 is upregulated in a variety of cancers, including breast cancer. High miR‐21 levels have been associated with poor prognosis. To determine the cellular localization of miR‐21 and to compare its expression levels with histopathological features, we performed in situ hybridization and semi‐quantitative assessment of the miR‐21 signal on 12 LN negative grade I (assumed low risk), and 12 LN positive grade II (high risk) breast cancers. miR‐21 was predominantly seen in cancer associated fibroblast‐like cells, with no difference in expression levels between grade I and grade II carcinomas. Immunohistochemical scoring of the prognostic proliferation marker Ki‐67 and tumor suppressor p53 showed that the miR‐21 expression levels significantly correlated with the Ki‐67 score (p = 0.043), whereas no correlation between p53 and miR‐21 was found. Our results indicate that miR‐21 may contribute to improve clinical stratification according to growth rate and facilitate tailored treatment of breast cancer patients.
Analysis of CD177 neutrophil expression in β‐thalassemia patientsMONTASER, LAILA MAHMOUD; EL‐RASHIDI, FARIDA HUSSEIN; ESSA, ENAS SAID; AZAB, SAMAR MOSTAFA
doi: 10.1111/j.1600-0463.2011.02755.xpmid: 21917004
Montaser LM, El‐Rashidi FH, Essa ES, Azab SM. Analysis of CD177 neutrophil expression in β‐thalassemia patients. APMIS 2011; 119: 674–80. Ineffective erythropoiesis plays a well established role in the pathophysiology of disease expression in β‐thalassemia major and intermedia. CD177 expression was investigated in different clinical conditions. The study aimed to analyze neutrophil expression of CD177 in β‐thalassemia patients and its correlation with serum soluble transferrin receptor (s‐TfR) concentration as a marker for the extent of erythropoiesis and hence disease severity in these patients. Flow cytometric analysis of neutrophil CD177 expression and enzyme immunoassay measurement of serum s‐TfR concentration were assessed in 45 β‐thalassemia patients of whom 36 had β‐thalassemia major and nine had β‐thalassemia intermedia. They were also assessed in 21 age‐ and gender‐matched control children. Neutrophil mean fluorescence intensity ratio (MFIR) of CD177 expression was significantly higher in patients than in controls (p < 0.001). There was highly significant increase in serum s‐TfR concentration in β‐thalassemia patients than in controls (p < 0.001). There was a highly significant positive correlation between MFIR of CD177 expression and serum s‐TfR concentration in β‐thalassemia patients (r = 0.59, p < 0.001). Elevated CD177 expression is not only a specific feature of polycythemia rubra vera (PV) but may be also an indicator of increased erythropoietic activity in thalassemia syndromes.
The use of Gram stain and matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry on positive blood culture: synergy between new and old technologyFUGLSANG‐DAMGAARD, DAVID; NIELSEN, CAMILLA HOULBERG; MANDRUP, ELISABETH; FUURSTED, KURT
doi: 10.1111/j.1600-0463.2011.02756.xpmid: 21917005
Fuglsang‐Damgaard D, Nielsen CH, Mandrup E, Fuursted K. The use of Gram stain and matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry on positive blood culture: synergy between new and old technology. APMIS 2011; 119: 681–88. Matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry (MALDI‐TOF‐MS) is promising as an alternative to more costly and cumbersome methods for direct identifications in blood cultures. We wanted to evaluate a simplified pre‐treatment method for using MALDI‐TOF‐MS directly on positive blood cultures using BacT/Alert blood culture system, and to test an algorithm combining the result of the initial microscopy with the result suggested by MALDI‐TOF‐MS. Using the recommended cut‐off score of 1.7 the best results were obtained among Gram‐negative rods with correct identifications in 91% of Enterobacteriaceae, 83% in aerobic/non‐fermentative Gram‐negative rods, whereas results were more modest among Gram‐positive cocci with correct identifications in 52% of Staphylococci, 54% in Enterococci and only 20% in Streptococci. Combining the results of Gram stain with the top reports by MALDI‐TOF‐MS, increased the sensitivity from 91% to 93% in the score range from 1.5 to 1.7 and from 48% to 85% in the score range from 1.3 to 1.5. Thus, using this strategy and accepting a cut‐off at 1.3 instead of the suggested 1.7, overall sensitivity could be increased from 88.1% to 96.3%. MALDI‐TOF‐MS is an efficient method for direct routine identification of bacterial isolates in blood culture, especially when combined with the result of the Gram stain.
Survival estimates after pancreatoduodenectomy skewed by non‐standardized histopathology reportsWESTGAARD, ARNE; CLAUSEN, OLE PETTER F.; GLADHAUG, IVAR P.
doi: 10.1111/j.1600-0463.2011.02783.xpmid: 21917006
Westgaard A, Clausen OPF, Gladhaug IP. Survival estimates after pancreatoduodenectomy skewed by non‐standardized histopathology reports. APMIS 2011; 119: 689–700. Survival estimates may be biased if quality control on histopathology is insufficient. We evaluated the effects of standardizing histopathology for pancreatoduodenectomy specimens and compared survival estimates based on standardized vs non‐standardized histopathological evaluation. Microscopic slides and histopathological reports from 311 consecutive pancreatoduodenectomies (1980–2004) were reviewed, including 104 adenocarcinomas (1980–1997) resected before and 123 adenocarcinomas (1998–2004) resected after standardizing histopathology. Histopathological factors were re‐evaluated for all primary adenocarcinomas (n = 227). The most frequent histological types were pancreatobiliary‐type (n = 145) and intestinal‐type (n = 73). Standardized histopathology was associated with sampling more blocks and nodes (p < 0.001), and with more frequent identification of non‐pancreatic tumour origin, nodal and margin involvement, perineural infiltration, and poor differentiation (p < 0.05). Standardized evaluation was necessary to discriminate between prognostic groups with respect to perineural infiltration and tumour size, but not to identify disparate prognostic subgroups with respect to nodal and margin involvement. Nodal involvement (N1 vs N0, p < 0.001) and histological type (pancreatobiliary vs intestinal, p < 0.001) were independent prognostic factors after pancreatoduodenectomy. Histopathological evaluation should be standardized to provide reliable prognostic estimates and to discriminate between prognostic subgroups. Lymph node involvement and histological type are independent prognostic factors after pancreatoduodenectomy for adenocarcinoma.
Secretory IgA–Fcα receptor interaction modulating phagocytosis and microbicidal activity by phagocytes in human colostrum of diabeticsFRANçA, EDUARDO LUZíA; MORCELI, GLILCIANE; FAGUNDES, DANNY LAURA GOMES; RUDGE, MARILZA VIEIRA CUNHA; CALDERON, IRACEMA DE MATTOS PARANHOS; HONORIO‐FRANçA, ADENILDA CRISTINA
doi: 10.1111/j.1600-0463.2011.02789.xpmid: 21917008
França EL, Morceli G, Fagundes DLG, Rudge MVC, Calderon I de MP, Honorio‐França AC. Secretory IgA–Fcα receptor interaction modulating phagocytosis and microbicidal activity by phagocytes in human colostrum of diabetics. APMIS 2011; 119: 710–19. The effects of secretory immunoglobulin A (SIgA) interaction with its specific Fcα receptors on colostral phagocytes needs further investigation, especially with respect to diabetic women. Accordingly, we studied the colostrum of hyperglycemic women to assess SIgA interactions with Fcα receptors of macrophages as well as the functional activity of these cells. The women were divided for colostrum sampling according to their glycemic status: normoglycemia (N = 51), mild hyperglycemia (N = 23), and diabetes (N = 25) groups. We determined the FcαR expression, the IgA on the surface and the surface‐bound IgA in colostrum macrophages. We also evaluated the superoxide release and bactericidal killing of these cells. Colostral phagocytes expressed FcαR, contained IgA on the surface and are able to bind to purified SIgA. The bactericidal activity of colostral phagocytes from the hyperglycemic women was similar to that of normoglycemic only when SIgA was used as opsonin. Addition of a MoAb anti‐human Fcα receptor resulted in a significant decrease of superoxide release and bacterial killing by macrophages when bacteria were opsonized with purified SIgA, suggesting an interaction between SIgA and FcαR. The stimulatory effects of SIgA on the functional activity of phagocytes therefore protect infants, especially of diabetic women, against intestinal infections.
Antimicrobial properties of natural coniferous rosin in the European Pharmacopoeia challenge testSIPPONEN, ARNO; LAITINEN, KIRSI
doi: 10.1111/j.1600-0463.2011.02791.xpmid: 21917009
Sipponen A, Laitinen K. Antimicrobial properties of natural coniferous rosin in the European Pharmacopoeia challenge test. APMIS 2011; 119: 720–24. Rosins (resins) are natural products of the coniferous trees. Purified rosin from the trunk of Norway spruce (Picea abies) is antibacterial against the gram‐positive bacteria, but not against the gram‐negative bacteria in agar plate diffusion test. In this study, we examined the antimicrobial properties of the coniferous rosin against bacteria and yeasts using the European Pharmacopoeia (EP) challenge test. The microbes tested were Staphylococcus aureus, methicillin‐resistant Staphylococcus aureus (MRSA), Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and Candida albicans. To prepare challenge media, purified rosin was mixed with a biologically inert salve in varying concentrations. The microbes were inoculated (5 × 105 microbes (bacteria) or 5 × 104 microbes (yeast, C.albicans)) into 10 g of the rosin‐containing challenge medium for 14 days at maximum. Samples were taken from the media for re‐cultivation of the microbes at time intervals of 1 h, 24 h, 4, 7, and 14 days. The microbicidal efficacy of the challenge media was estimated by reduction of the number of the colony forming units (CFU) of microbes in the test samples. A reduction of more than 103 CFU for bacteria and 102 CFU for fungi in 7 days was considered to indicate a significant microbicidal action. Pure rosin was antimicrobial within 24 h against all microbes tested. The 0.5% rosin‐salve medium (w/w) did not differ in microbicidal effects from the rosin‐free salve medium (control). A raise of the rosin concentration resulted in increase of the microbicidal effect of the rosin‐salve medium against all micro‐organisms tested. Rosin concentration of 10% (w/w) in the medium significantly reduced the colonization of S. aureus (including MRSA) within 24 h and significantly reduced the colonization of all other micro‐organisms within 4 days. Rosin is strongly microbicidal against a wide range of microbes, against both gram‐positive and gram‐negative bacteria, and against C. albicans, in the EP challenge test. The minimum concentration of rosin is 10% (w/w) to prevent the preservation of the microbes in the rosin‐salve media.
VIM and IMP metallo‐β‐lactamases and other extended‐spectrum β‐lactamases in Escherichia coli and Klebsiella pneumoniae from environmental samples in a Tunisian hospitalCHOUCHANI, CHEDLY; MARRAKCHI, RIM; FERCHICHI, LEILA; EL SALABI, ALLAAEDDIN; WALSH, TIMOTHY R.
doi: 10.1111/j.1600-0463.2011.02793.xpmid: 21917010
Chouchani C, Marrakchi R, Ferchichi L, El Salabi A, Walsh TR. VIM and IMP metallo‐β‐lactamases and other extended‐spectrum β‐lactamases in Escherichia coli and Klebsiella pneumoniae from environmental samples in a Tunisian hospital. APMIS 2011; 119: 725–32. An extremely drug‐resistant Enterobacteriaceae species emerged in Kasserine Hospital, Tunisia between 2009 and 2010 causing a local outbreak. We aimed to characterize extended‐spectrum β‐lactamase (ESBL) and metallo‐β‐lactamase (MBL)‐producing Enterobacteriaceae from the hospital environment. Swabs were collected from ten different wards from Kasserine Hospital, Tunisia. A total of 46 isolates were cultured onto MacConkey agar supplemented with ceftazidime to select for ESBL‐producing Enterobacteriaceae. Identification and susceptibility patterns were performed using Phoenix‐automated phenotypic identification criteria. Extended spectrum β‐lactamases (ESBLs) were detected using cefepime ESBL E‐test. Colony blotting was first used to detect the occurrence of blaSHV, blaCTX‐M, blaCMY, blaIMP, and blaVIM genes. PCR was used to amplify these genes, and the amplicons were sequenced and analyzed. Total DNA was digested with XbaI, and PFGE was used to type the major isolates that produced IMP‐1. Among the 46 isolates, 63% were Klebsiella pneumoniae, 13% were Escherichia coli, 8.7% were Proteus mirabilis, 6% were Enterobacter cloaceae, 4.3% were Providencia rettgeri, 2.5% were Serratia marcescens, and 2.5% were Pantoea agglomerans. PCR amplification and DNA sequencing showed that hospital environment isolates produced SHV‐125, CTX‐M‐15, CMY‐2 ESBLs, and IMP‐1 and VIM‐2 MBLs. PFGE typing showed the emergence of IMP‐1 MBL‐producing K. pneumoniae isolates that were not clonal. In this study, we report the first characterization of IMP‐1 and VIM‐2 MBL‐producing K. pneumoniae and E. coli isolates collected from Kasserine Hospital, Tunisia.