ABSTRACT
Burns can cause skin damage, facilitating the entry of fungi and other microorganisms into the body, leading to infections. Fusarium is a fungus capable of infecting individuals with burn injuries. Diagnosing and treating Fusarium infections in burn patients can be challenging due to the manifestation of nonspecific symptoms. This study aims to investigate case reports and case series from published literature describing Fusarium infection in burned patients, in order to assess treatment regimens, clinical outcomes, and make recommendations for future management. We conducted searches on Web of Science, PubMed, ScienceDirect, and Medline for all case reports and case series containing keywords 'Burn', 'Burns', 'Burned', 'Fusarium', or 'Fusariosis' in the title or abstract. All burn patients who developed Fusarium fungal infections between January 1974 and March 2023 were included in the study. Demographic and clinical data were analyzed retrospectivity. The final analysis incorporates 24 case reports encompassing a total of 87 burn patients with Fusarium infection. Patient ages ranged from one to 85 years, with the majority being male (53%). The median percentage of burn surface area was 78%, and the skin in the face, upper limbs, and lower limbs were the most commonly infected sites. Fungal infections appeared around 10 days after the burn injury on average. The majority of the patients were identified through culture or histopathology. The Fusarium dimerum species complex, which was found in nine patients, was the most frequently identified Fusarium species complex. Amphotericin B was the most preferred treatment drug, followed by voriconazole, and 62% of patients underwent debridement. In our study, 23 patients (37%) died from fungal infections. Implementing early and effective treatment protocols targeting Fusarium spp. in burn treatment units can significantly reduce mortality rates. It is critical to enhance the understanding of fusariosis epidemiology and emphasize the importance of maintaining a high clinical suspicion for this condition in burn patients.
Subject(s)
Antifungal Agents , Burns , Fusariosis , Fusarium , Humans , Fusariosis/microbiology , Fusariosis/drug therapy , Burns/complications , Burns/microbiology , Antifungal Agents/therapeutic use , Fusarium/isolation & purification , Male , Adult , Middle Aged , Female , Aged , Young Adult , Aged, 80 and over , Adolescent , Child , Child, Preschool , InfantABSTRACT
The intraductal oncocytic papillary neoplasm (IOPN) of the pancreas has been recognized by WHO classification as a unique intraductal papillary mucinous neoplasm (IPMN) category. IOPN is composed of oxyphil cells, usually expressing MUC5AC, MUC6, and Hep Par-1, and harboring PRKACA/B fusion genes as their genetic hallmark. Although IOPNs are associated with an infiltrative adenocarcinoma in up to 30% of cases, the survival rate after surgical resection approaches 100%. This highlights the importance of the correct IOPN diagnosis, above all in cases with an associated invasive component. In this study, the immunohistochemical expression of CD117 was investigated in 111 IPMNs, including 17 oncocytic, 45 gastric, 20 pancreatico-biliary, and 29 intestinal IPMNs. We also tested the expression of MUC5AC, MUC6, and Hep Par-1 in the IOPN cohort. CD117 positivity was significantly more frequent in IOPNs compared to the other IPMN subtypes (p < 0.0001). Furthermore, within IOPN, a lower or absent CD117, MUC5AC, MUC6, and Hep Par-1 expression tended to be associated with the presence of an infiltrative component. Our findings shed light into the biology of these complex lesions, which are confirmed to be a distinctive IPMN subtype; notably, CD117 emerged as a potential, additional tool in the differential diagnosis of IPMNs.
Subject(s)
Adenocarcinoma, Mucinous/pathology , Biomarkers, Tumor/metabolism , Carcinoma, Papillary/pathology , Oxyphil Cells/pathology , Pancreatic Intraductal Neoplasms/pathology , Proto-Oncogene Proteins c-kit/metabolism , Adenocarcinoma, Mucinous/metabolism , Carcinoma, Papillary/metabolism , Cell Line, Tumor , Cohort Studies , Humans , Mucins/metabolism , Pancreatic Intraductal Neoplasms/metabolismABSTRACT
Undifferentiated carcinoma of the pancreas with osteoclast-like giant cells (UCOGC) is currently considered a morphologically and clinically distinct variant of pancreatic ductal adenocarcinoma (PDAC). In this study, we report clinical and pathological features of a series of 22 UCOGCs, including the whole exome sequencing of eight UCOGCs. We observed that 60% of the UCOGCs contained a well-defined epithelial component and that patients with pure UCOGC had a significantly better prognosis than did those with an UCOGC with an associated epithelial neoplasm. The genetic alterations in UCOGC are strikingly similar to those known to drive conventional PDAC, including activating mutations in the oncogene KRAS and inactivating mutations in the tumor suppressor genes CDKN2A, TP53, and SMAD4. These results further support the classification of UCOGC as a PDAC variant and suggest that somatic mutations are not the determinants of the unique phenotype of UCOGC. Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Subject(s)
Carcinoma, Pancreatic Ductal/pathology , Pancreatic Neoplasms/pathology , Aged , Aged, 80 and over , Carcinoma, Pancreatic Ductal/genetics , Carcinoma, Pancreatic Ductal/mortality , Exome/genetics , Female , Humans , Immunohistochemistry , Male , Middle Aged , Mutation/genetics , Neoplasm Proteins/genetics , Osteoclasts/pathology , Pancreatic Neoplasms/genetics , Pancreatic Neoplasms/mortality , Pancreatic NeoplasmsABSTRACT
Amphicrine neoplasms (ANs) are poorly understood epithelial malignancies composed of cells with co-existing exocrine-neuroendocrine features. Here, we report a recurrent mucin-producing gastric amphicrine tumor co-expressing neuroendocrine (chromogranin-A, synaptophysin, and CD56) and pancreatic acinar cell (BCL10 and trypsin) markers, arisen in a 64-year-old woman during adjuvant immunotherapy for melanoma. Ki-67 was < 2%. The gastric background context was atrophic gastritis. Next-generation sequencing showed MEN1 mutation (p.P71fs*42) coupled with loss of heterozygosity. The key lessons were as follows: (1) gastric ANs can show the co-existence of exocrine mucin-producing elements with neuroendocrine and pancreatic acinar differentiation; (2) they may represent a new entity arising in the context of atrophic gastritis and during immunotherapy; (3) they should be considered in the diagnostic workup of gastric neuroendocrine tumors; and (4) their molecular profile can show striking similarities with well-differentiated neuroendocrine tumors. These findings may be of help to improve the knowledge and the biological taxonomy of ANs.
Subject(s)
Gastritis, Atrophic , Neuroendocrine Tumors , Pancreatic Neoplasms , Stomach Neoplasms , Female , Humans , Middle Aged , Acinar Cells/pathology , Neoplasm Recurrence, Local , Stomach Neoplasms/genetics , Stomach Neoplasms/therapy , Stomach Neoplasms/diagnosis , Neuroendocrine Tumors/therapy , Neuroendocrine Tumors/diagnosis , Cell Differentiation , Mucins , Pancreatic Neoplasms/genetics , Pancreatic Neoplasms/therapy , Pancreatic Neoplasms/diagnosis , Biomarkers, Tumor/geneticsABSTRACT
BACKGROUND: Intestinal neuroendocrine tumours (I-NETs) represent a non-negligible entity among intestinal neoplasms, with metastatic spreading usually present at the time of diagnosis. In this context, effective molecular actionable targets are still lacking. Through transcriptome analysis, we aim at refining the molecular taxonomy of I-NETs, also providing insights towards the identification of new therapeutic vulnerabilities. MATERIALS AND METHODS: A retrospective series of 38 primary sporadic, surgically-resected I-NETs were assessed for transcriptome profiling of 20,815 genes. RESULTS: Transcriptome analysis detected 643 highly expressed genes. Unsupervised hierarchical clustering, differential expression analysis and gene set enriched analysis identified three different tumour clusters (CL): CL-A, CL-B, CL-C. CL-A showed the overexpression of ARGFX, BIRC8, NANOS2, and SSTR4 genes. Its most characterizing signatures were those related to cell-junctions, and activation of mTOR and WNT pathway. CL-A was also enriched in T CD8 + lymphocytes. CL-B showed the overexpression of PCSK1, QPCT, ST18, and TPH1 genes. Its most characterizing signatures were those related to adipogenesis, neuroendocrine metabolism, and splice site machinery-related processes. CL-B was also enriched in T CD4 + lymphocytes. CL-C showed the overexpression of ALB, ANG, ARG1, and HP genes. Its most characterizing signatures were complement/coagulation and xenobiotic metabolism. CL-C was also enriched in M1/2 macrophages. These CL-based differences may have therapeutic implications in refining the management of I-NET patients. At last, we described a specific gene-set for differentiating I-NET from pancreatic NET. DISCUSSION: Our data represent an additional step for refining the molecular taxonomy of I-NET, identifying novel transcriptome subgroups with different biology and therapeutic opportunities.
Subject(s)
Intestinal Neoplasms , Neuroendocrine Tumors , Humans , Neuroendocrine Tumors/genetics , Neuroendocrine Tumors/pathology , Retrospective Studies , Gene Expression Profiling , Intestines/pathology , Transcriptome , Intestinal Neoplasms/genetics , Intestinal Neoplasms/pathologyABSTRACT
Intraductal oncocytic papillary neoplasm (IOPN) of the pancreas is a distinct entity from intraductal papillary mucinous neoplasms (IPMNs) and is considered one of the precursor lesions of pancreatic cancer. Through immunohistochemistry (IHC) and an artificial intelligence (AI)-based approach, this study aims at characterizing its immune microenvironment. Whole-slide IHC was performed on a cohort of 15 IOPNs, 2 of which harboring an associated adenocarcinoma. The following markers were tested: CD3, CD4, CD8, CD20, CD68, CD163, PD-1, PD-L1, MLH1, PMS2, MSH2, and MSH6. The main findings can be summarized as follows: (i) CD8+ T lymphocytes were the predominant immune cells (p < 0.01); (ii) the vast majority of macrophages were concurrently CD68+ and CD163+; (iii) all tumors showed an activated PD-1/PD-L1 axis, but none had mismatch repair deficiency; (iv) AI-based analysis revealed the presence of 2 distinct regions in each case, namely, Re1, localized at the center of the tumor, and Re2, located at tumor periphery; (v) the infiltrating component of the 2 invasive IOPNs showed a smaller extent of Re1 and a reduced rate of CD4+ cells, as well as a larger extent of Re2 and increased rate of CD8+ cells. IOPNs are lesions enriched in immune cells, with a predominance of CD8+ T lymphocytes and class 2 macrophages. Differently from IPMN-oncogenesis, the progression towards invasive carcinoma is accompanied by an increased rate of CD8+ lymphocytes. This finding may suggest the presence of an active self-immune surveillance in invasive IOPNs, potentially explaining, at least in part, the excellent survival rate of IOPN patients.
Subject(s)
Carcinoma, Pancreatic Ductal , Pancreatic Neoplasms , Humans , Carcinoma, Pancreatic Ductal/pathology , B7-H1 Antigen , Artificial Intelligence , Programmed Cell Death 1 Receptor , Pancreatic Neoplasms/pathology , Pancreas/pathology , Tumor MicroenvironmentABSTRACT
Hepatoid tumors (HTs) represent a rare group of neoplasms that are histologically similar to hepatocellular carcinoma but arise outside the liver. The current World Health Organization classification recognizes the hepatoid morphology of pancreatic tumors only as a possible variant of pancreatic ductal adenocarcinoma (PDAC). Here, we describe two cases of "pure" HT of the pancreas showing common features and characterized by indolent biological behavior. These tumors were roundish nodules with pushing borders, hyaline globules, and pure hepatoid histology; they were diffusely positive for ß-catenin and LEF1 on immunohistochemistry. At next-generation sequencing, both neoplasms harbored only one pathogenic somatic mutation that affected the CTNNB1 gene at exon 3 and showed a loss of heterozygosity on chromosomes 18 and 21. By integrating macroscopic and microscopic features, along with their molecular profiles, we advocate that such tumors represent a distinct entity from PDAC and should be considered a new variant of solid pseudopapillary neoplasms. The recognition of this new neoplastic category may have immediate implications not only for tumor taxonomy but also for clinical practice.
Subject(s)
Carcinoma, Pancreatic Ductal , Liver Neoplasms , Pancreatic Neoplasms , beta Catenin , Biomarkers, Tumor/genetics , Carcinoma, Pancreatic Ductal/genetics , Carcinoma, Pancreatic Ductal/pathology , Humans , Liver Neoplasms/genetics , Liver Neoplasms/pathology , Mutation/genetics , Pancreas/pathology , Pancreatic Neoplasms/genetics , Pancreatic Neoplasms/pathology , beta Catenin/genetics , Pancreatic NeoplasmsABSTRACT
Juvenile polyposis (JP) is a rare familial syndrome characterized by the development of numerous hamartomatous polyps of the gastrointestinal tract and by an increased risk of developing gastrointestinal cancers. It follows a pattern of autosomal dominant inheritance and is associated with germline variants of SMAD4 or BMPR1A genes. Differential diagnosis may be difficult based on histology alone, due to morphological similarities to other familial syndromes. Here we report a case of familial JP diagnosed in a 50-years woman with a familial history positive for gastrointestinal cancers and other tumor types. The patient presented with severe iron deficiency anemia and showed numerous polyps in the stomach and jejunum according to endoscopy and imaging. She underwent an intra-gastric laparoscopic removal of the major gastric polyp, followed by jejunal exploration and resection of a segment with multiple neoformations. Histological examination revealed the presence of hamartomatous polyposis. Gastric and intestinal samples were analyzed with next-generation sequencing. Molecular analysis showed that the patient harbored a germline splicing site variant of SMAD4, c.1139 + 3A > G, which was complemented by different somatic variants of the same gene in the different polyps. Immunohistochemistry for SMAD4 confirmed loss of protein expression in the polyps, with regular expression in normal cells. cDNA sequencing further confirmed the findings. We thus definitively diagnosed the woman as having JP thanks to an integrated approach based on histology, immunohistochemistry and molecular analysis. The identified variants, all previously reported as variants of unknown significance, were classified as pathogenic as they complemented each other leading to SMAD4 loss.
Subject(s)
Adenomatous Polyps , Gastrointestinal Neoplasms , Intestinal Polyposis , Neoplastic Syndromes, Hereditary , Polyps , Female , Humans , Intestinal Polyposis/diagnosis , Intestinal Polyposis/genetics , Intestinal Polyposis/pathology , Neoplastic Syndromes, Hereditary/diagnosis , Neoplastic Syndromes, Hereditary/genetics , Smad4 Protein/genetics , Adenomatous Polyps/genetics , Polyps/genetics , Gastrointestinal Neoplasms/genetics , SyndromeABSTRACT
Undifferentiated carcinoma (UC) and undifferentiated carcinoma with osteoclast-like giant cells (UCOGC) are peculiar variants of pancreatic ductal adenocarcinoma (PDAC), characterized by hypercellularity and absence of glandular patterns. The inflammatory microenvironment is peculiar in UCOGC, since it is dominated by macrophages and osteoclast-like giant cells. However, from a molecular point of view, both UC and UCOGC are very similar to conventional PDAC, sharing alterations of the most common genetic drivers. Clinically, UC usually show a worse prognosis, whereas UCOGC may show a better prognosis if it is not associated with a PDAC component. To highlight potential biological differences between these entities, we investigated the role of the epithelial to mesenchymal transition (EMT) in UC and UCOGC. Specifically, we analyzed the immunohistochemical expression of three well-known EMT markers, namely Twist1, Snai2, and E-cadherin, in 16 cases of UCOGC and 10 cases of UC. We found that EMT is more frequently activated in UC (10/10 cases) than in UCOGC (8/16 cases; p = 0.05). Furthermore, in UCOGC, EMT was activated with a higher frequency in cases with an associated PDAC component. Snai2 was the most frequently and strongly expressed marker in both tumor types (10/10 UC, 8/16 UCOGC), and its expression was higher in UC than in UCOGC (mean immunohistochemical score: 4.8 in UC vs. 2.1 in UCOGC, p < 0.01). Our results shed new light on the biology of UC and UCOGC: EMT appeared as a more important process in UC, and Snai2 emerged as a central EMT effector in this setting.
Subject(s)
Cell Differentiation , Epithelial-Mesenchymal Transition , Osteoclasts/pathology , Pancreatic Neoplasms/pathology , Antigens, CD/analysis , Baltimore , Biomarkers, Tumor/analysis , Cadherins/analysis , Humans , Immunohistochemistry , Italy , Neoplasm Staging , Nuclear Proteins/analysis , Osteoclasts/chemistry , Pancreatic Neoplasms/chemistry , Pancreatic Neoplasms/therapy , Retrospective Studies , Snail Family Transcription Factors/analysis , Twist-Related Protein 1/analysisABSTRACT
CD200 has been recently indicated as a robust marker of well-differentiated neuroendocrine neoplasms. Here, we evaluate its role in differential diagnosis of solid pancreatic neoplasms. We immunostained for CD200 22 solid pseudopapillary neoplasms (SPNs), 8 acinar carcinomas (ACs), 2 pancreatoblastomas (PBs), 138 neuroendocrine tumors (PanNETs), and 48 ductal adenocarcinomas. All SPNs showed strong cytoplasmic and membranous staining for CD200, while only one case of AC had focal positivity. The two PBs showed focal CD200 positivity, mainly located in squamoid nests. The vast majority of PanNETs (96%) showed strong cytoplasmic and membranous staining for CD200, whereas all PDACs were negative. As both PanNETs and SPNs express CD200, it has no role in the differential diagnosis between these two entities.
Subject(s)
Antigens, CD/analysis , Biomarkers, Tumor/analysis , Carcinoma, Papillary/immunology , Neuroendocrine Tumors/immunology , Pancreatic Neoplasms/immunology , Carcinoma, Acinar Cell/immunology , Carcinoma, Acinar Cell/pathology , Carcinoma, Pancreatic Ductal/immunology , Carcinoma, Pancreatic Ductal/pathology , Carcinoma, Papillary/pathology , Diagnosis, Differential , Humans , Immunohistochemistry , Neuroendocrine Tumors/pathology , Pancreatic Neoplasms/pathology , Predictive Value of TestsABSTRACT
Undifferentiated carcinoma with osteoclast-like giant cells (UCOGC), a variant of pancreatic ductal adenocarcinoma (PDAC), has a striking genetic similarity to PDAC but a significantly improved overall survival. We hypothesize that this difference could be due to the immune response to the tumor, and as such, we investigated the expression of PD-1, PD-L1, and CD163 in a series of UCOGC. To this aim, 27 pancreatic UCOGCs (11 pure and 16 PDAC-associated), 5 extrapancreatic tumors with osteoclast-like giant cells and 10 pancreatic anaplastic carcinomas were immunostained using antibodies against PD-1, PD-L1, and CD163. In pancreatic UCOGCs, PD-L1 was expressed in neoplastic cells of 17 (63%) of 27 cases, more often in cases with an associated PDAC (Pâ¯=â¯.04). Expression of PD-L1 was associated with poor prognosis, confirmed by multivariate analysis: patients with PD-L1-positive UCOGCs had a risk of all-cause mortality that was 3 times higher than did patients with PD-L1-negative UCOGCs (hazard ratio, 3.397; 95% confidence interval, 1.023-18.375; Pâ¯=â¯.034). PD-L1 expression on tumor cells was also associated with aberrant P53 expression (Pâ¯=â¯.035). PD-1 was expressed on rare lymphocytes in 12 UCOGCs (44.4%), mainly located at the tumor periphery. CD163 was expressed on histiocytes, with a diffuse and strong staining pattern in all UCOGCs. Extrapancreatic tumors with osteoclast-like giant cells showed very similar staining patterns for the same proteins. Anaplastic carcinomas have some similarities to UCOGCs, but PD-L1 has no prognostic roles. Our results may have important implications for immunotherapeutic strategies in UCOGCs; these tumors may also represent a model for future therapeutic approaches against PDAC.
Subject(s)
Antigens, CD/analysis , Antigens, Differentiation, Myelomonocytic/analysis , B7-H1 Antigen/analysis , Biomarkers, Tumor/analysis , Carcinoma, Pancreatic Ductal/immunology , Cell Differentiation , Giant Cells/immunology , Osteoclasts/immunology , Pancreatic Neoplasms/immunology , Programmed Cell Death 1 Receptor/analysis , Receptors, Cell Surface/analysis , Adult , Aged , Aged, 80 and over , Carcinoma, Pancreatic Ductal/mortality , Carcinoma, Pancreatic Ductal/pathology , Carcinoma, Pancreatic Ductal/surgery , Europe , Female , Giant Cells/pathology , Histiocytes/chemistry , Histiocytes/pathology , Humans , Immunohistochemistry , Indiana , Lymphocytes/immunology , Lymphocytes/pathology , Male , Middle Aged , Neoplasm Staging , Osteoclasts/pathology , Pancreatic Neoplasms/mortality , Pancreatic Neoplasms/pathology , Pancreatic Neoplasms/surgery , PhenotypeABSTRACT
BACKGROUND AND AIM OF THE WORK: Idiopathic Pulmonary Fibrosis/Usual Interstitial Pneumonia (IPF/UIP) is a diffuse and progressive lung disease whose pathogenesis is incompletely understood. Recently, the presence of Herpesvirus-specific DNA was detected in the large majority of cases of a series of IPF/UIP and other lung interstitial diseases. We have therefore tested our own IPF/UIP series for the presence of HHV-8 and EBV proteins. METHODS: We used a variety of sensitive technologies including immunohistochemistry with NCL-HHV8-LNA and EBV-LMP1 antibodies, as well as of EBV RNA (EBER) by in-situ hybridisation; the presence of HHV-8 and EBV DNA was also investigated by means of PCR and subsequent analysis using a microfluidic apparatus. RESULTS: Despite the good sample quality, immunohistochemical, in-situ hybridisation and PCR results were negative for both EBV and HHV-8. CONCLUSIONS: We conclude that EBV and HHV-8 are not involved in the pathogenesis of IPF/UIP.
Subject(s)
Antibodies, Viral/analysis , DNA, Viral/analysis , Herpesvirus 4, Human , Herpesvirus 8, Human , Pulmonary Fibrosis/virology , Adult , Aged , Diagnosis, Differential , Epstein-Barr Virus Infections/diagnosis , Epstein-Barr Virus Infections/virology , Female , Herpesviridae Infections/diagnosis , Herpesviridae Infections/virology , Herpesvirus 4, Human/genetics , Herpesvirus 4, Human/immunology , Herpesvirus 8, Human/genetics , Herpesvirus 8, Human/immunology , Humans , Immunohistochemistry , In Situ Hybridization , Lung Diseases, Interstitial/diagnosis , Lung Diseases, Interstitial/virology , Male , Middle Aged , Polymerase Chain Reaction , Pulmonary Fibrosis/pathologyABSTRACT
p63, a member of the p53 family, is involved in the survival and differentiation of reserve/stem cells in different epithelia. To unveil the possible role of p63 in thymic physiology and pathology, we investigated the expression of p63 isoforms in normal thymus, thymomas and other mediastinal tumours. All samples were analysed using immunohistochemistry with three different antibodies: 4A4 antibody recognising all p63 isoforms, p40 antibody reacting only with truncated dominant-negative isoforms (DeltaN-p63) and H-129 antibody recognising all alpha-isoforms. Reverse-transcription polymerase chain reaction (RT-PCR), and real-time PCR analyses were performed on RNA extracted from frozen samples of four thymomas and two primary-mediastinal large-B-cell lymphoma (PMLBCL). In normal thymus, DeltaN-p63alpha was expressed in all cortical and medullary epithelial cells, with decreasing intensity in Hassall's corpuscles. This phenotype was conserved in neoplastic transformation since all 54 investigated thymomas (World Health Organization types A, AB, B1, B2, B3, C) expressed DeltaN-p63alpha (virtually 100% cells). The predominance of DeltaN-p63alpha isoform mRNA was confirmed by real-time PCR. Among other mediastinal tumours, DeltaN-p63alpha was only expressed in those displaying either a stratified epithelial component (teratomas) or epidermoid differentiation (lung carcinoma). Among lymphomas, T-cell-precursor lymphomas did not express p63, whereas most PMLBCL expressed TA-p63alpha (7/8).
Subject(s)
DNA-Binding Proteins/metabolism , Thymoma/metabolism , Thymus Gland/metabolism , Thymus Neoplasms/metabolism , Cell Nucleus/metabolism , Cell Nucleus/pathology , DNA-Binding Proteins/genetics , DNA-Binding Proteins/immunology , Fluorescent Antibody Technique, Indirect , Humans , Mediastinal Neoplasms/genetics , Mediastinal Neoplasms/metabolism , Mediastinal Neoplasms/pathology , Protein Isoforms/genetics , Protein Isoforms/immunology , Protein Isoforms/metabolism , RNA, Messenger/metabolism , RNA, Neoplasm/analysis , Reverse Transcriptase Polymerase Chain Reaction , Thymoma/pathology , Thymus Gland/cytology , Thymus Gland/pathology , Thymus Neoplasms/pathologyABSTRACT
The BACTEC MGIT system (M960), a fully automated, non radiometric instrument, designed for rapid detection of acid fast bacilli (AFB) from clinical specimens, was compared with the radiometric BACTEC 460 system (B460) and Löwenstein-Jensen (L-J) solid medium. A total of 1,093 respiratory and extrapulmonary specimens were decontaminated by the NALC-NaOH standard method, and randomly inoculated into the media. A total of 122 mycobacteria were recovered, including 47 Mycobacterium tuberculosis complex (MTB) isolates and 75 nontuberculous mycobacteria (NTM) isolates. Overall recovery rates were 59% for M960 system (p < 0.0001), 58.2% for L-J. medium (p < 0.0001) and 82% for Bactec 460 system, whereas rates for MTB alone were 91.5, 76.6 (p = 0.007), and 95.7%, respectively. The combination of M960 or B460 systems with L-J medium showed the same recovery rates for MTB strains (97.9%), whereas NTM rates were 68% (p < 0.0001) and 93.3%, respectively. Mean time to detection of smear-positive MTB, smear-negative MTB, and NTM were 12.2, 13.4, and 23.3 days, respectively, with the M960, 11.7, 21.3, and 24.8 days with the B460 and 20.4, 28.7, and 28.4 days with L-J medium. The M960 system showed a contamination rate of 9.8%, while B460 and L-J medium showed contamination rates of 4.3 and 3.8% respectively. In conclusion, the M960 system appeared to be accurate and rapid for the recovery of MTB, but reduced recovery of NTM and a high number of contaminated cultures deserve further study in order to assess if this system can represent a valuable alternative to the radiometric system.
Subject(s)
Bacteriological Techniques , Mycobacterium tuberculosis/isolation & purification , Reagent Kits, Diagnostic , Tuberculosis/diagnosis , Culture Media , Humans , Probability , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
The clonal/neoplastic nature of Langerhans cell histiocytosis (LCH) has recently been demonstrated by a high prevalence of BRAF mutations, including pulmonary LCH (PLCH). We hypothesized that BRAF-induced senescence, as demonstrated in nevi and melanoma, is involved in the pathogenesis of LCH and PLCH. In a series of pulmonary (19 cases) and non-pulmonary LCH (19 cases), including five aggressive cases, we investigated occurrence of the BRAF V600E mutation by molecular analysis and/or immunohistochemistry using a validated antibody (VE1). The expression of cell-senescence markers p16(INK4a) and p21(CIP1/WAF1) was also immunohistochemically investigated. We demonstrated that 6/19 cases of LCH and 12/19 cases of PLCH were VE1 positive, matching with molecular analysis, and in all cases both p16(INK4a) and p21(CIP1/WAF1) were expressed, irrespective of BRAF mutation status. Interestingly, all the aggressive cases did not express p16(INK4a), thus suggesting that loss of senescence control could be related to clinical aggressiveness of LCH, as in melanoma.
Subject(s)
Cellular Senescence/genetics , Histiocytosis, Langerhans-Cell/genetics , Lung/metabolism , Mutation, Missense , Proto-Oncogene Proteins B-raf/genetics , Adolescent , Adult , Aged , Child, Preschool , Cyclin-Dependent Kinase Inhibitor p16/metabolism , Cyclin-Dependent Kinase Inhibitor p21/metabolism , DNA Mutational Analysis , Female , Histiocytosis, Langerhans-Cell/metabolism , Histiocytosis, Langerhans-Cell/pathology , Humans , Immunohistochemistry , Infant , Lung/pathology , Male , Middle Aged , Proto-Oncogene Proteins B-raf/metabolism , Young AdultABSTRACT
AIMS: The macro- and microcirculation disease, in patients with type 2 diabetes mellitus (T2DM), induces ischemic wounds of the lower limbs. We have tried to reduce the aggregation of red blood cells and to improve the O2 supply to the tissues and speed the healing of ulcers in T2DM patients. METHODS: We enrolled 25 obese subjects without glucose intolerance (group A; BMI greater than 30 kg/m2), 20 obese adults intolerant to glucose (group B) and two subgroups, groups C and D, with T2DM and with leg ulcers. The groups A, B and C were treated with PESF. Body weight, O2 extraction, the capillary pulse, blood pressure and the surface of the ulcers were monitored. RESULTS: The technique PESF shows to have positive effects on the metabolism, on the reduction of body weight in the groups A and B, increasing extraction of O2 in group C and increase the speed of healing of wounds in group C compared to group D. In group A, there was a significant reduction in systolic and diastolic blood pressure. CONCLUSIONS: The technique PESF has affected the metabolic processes and the speed of wound healing ulcer in patients with T2DM.
Subject(s)
Diabetes Mellitus, Type 2/complications , Diabetic Foot/therapy , Electric Stimulation Therapy/methods , Foot/blood supply , Ischemia/therapy , Static Electricity , Wound Healing , Aged , Biomarkers/blood , Blood Pressure , Diabetes Mellitus, Type 2/blood , Diabetes Mellitus, Type 2/diagnosis , Diabetic Foot/blood , Diabetic Foot/diagnosis , Diabetic Foot/etiology , Glycated Hemoglobin/metabolism , Humans , Ischemia/blood , Ischemia/diagnosis , Ischemia/etiology , Italy , Male , Middle Aged , Obesity/complications , Obesity/diagnosis , Oxygen/blood , Regional Blood Flow , Time Factors , Treatment OutcomeABSTRACT
The performance of the fully automated BACTEC MGIT 960 (M960) system for the testing of Mycobacterium tuberculosis susceptibility to streptomycin (SM), isoniazid (INH), rifampin (RMP), ethambutol (EMB), and pyrazinamide (PZA) was evaluated with 100 clinical isolates and compared to that of the radiometric BACTEC 460TB (B460) system. The agar proportion method and the B460 system were used as reference methods to resolve the discordant results for SM, INH, RMP, and EMB (a combination known as SIRE) and PZA, respectively. The overall agreements were 96.3% for SIRE and 92% for PZA. For SIRE, a total of 26 discrepancies were found and were resolved in favor of the M960 system in 8 cases and in favor of the B460 system in 18 cases. The M960 system produced 8 very major errors (VME) and 10 major errors (ME), while the B460 system showed 4 VME and 4 ME. No statistically significant differences were found. Both systems exhibited excellent performance, but a higher number of VME was observed with the M960 system at the critical concentrations of EMB and SM. For PZA, a total of eight discrepancies were observed and were resolved in favor of the M960 system in one case and in favor of the B460 system in seven cases; no statistically significant differences were found. The M960 system showed four VME and three ME. The mean times to report overall PZA results and resistant results were 8.2 and 9.8 days, respectively, for the M960 system and 7.4 and 8.1 days, respectively, for the B460 system. Statistically significant differences were found. The mean times to report SIRE results were 8.3 days for the M960 system and 8.2 days for the B460 system. No statistically significant differences were found. Twelve strains tested for SIRE susceptibility and seven strains tested for PZA susceptibility had been reprocessed because of contamination. In conclusion, the M960 system can represent a valid alternative to the B460 for M. tuberculosis susceptibility testing; however, the frequent contamination of the tests needs to be improved.
Subject(s)
Antitubercular Agents/pharmacology , Microbial Sensitivity Tests/methods , Mycobacterium tuberculosis/drug effects , Automation/methods , Humans , Mycobacterium tuberculosis/isolation & purification , Radiometry/methods , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
DipStreak is a new urine culture device with two types of agar attached back-to-back on a plastic paddle. It combines dip-slide technology and an original streaking inoculation mechanism, allowing for bacterial counting and colony isolation. The performance of the DipStreak device with two different medium formulations, CHROMagar and MacConkey media in study A and UriSelect 3 and MacConkey media in study B, was evaluated and compared to that of the reference streak method by using plates of cystine-lactose-electrolyte-deficient (CLED) agar, tryptic soy agar with 5% sheep blood, and UriSelect 3 medium. In study A, 2,000 urine specimens were processed and 511 cultures were found positive. The DipStreak device and the UriSelect 3 and CLED medium plates gave the same detection rate, 99.7%. For the direct identification of Escherichia coli, Proteus mirabilis, and Enterococcus sp. isolates, the DipStreak device and the UriSelect 3 medium plate showed overall sensitivities of 97 and 93.4%, respectively. In study B, 3,000 urine specimens were processed and 714 cultures were found positive. The DipStreak device and the UriSelect 3 and CLED medium plates gave detection rates of 99.4, 99.9, and 99.2%, respectively. For the direct identification of E. coli, P. mirabilis, and Enterococcus sp. isolates, the DipStreak device and the UriSelect 3 medium plate showed overall sensitivities of 88 and 94.4%, respectively. In conclusion, the DipStreak device with both medium formulations represents an attractive and excellent screening method for the reliable detection, counting, and presumptive identification of urinary tract pathogens. It enables bedside urine inoculation and provides a valid means of transporting the sample back to the laboratory, decreasing drastically the rate of false-positive results due to bacterial overgrowth and reducing associated costs.
Subject(s)
Candida/isolation & purification , Gram-Negative Bacteria/isolation & purification , Gram-Positive Bacteria/isolation & purification , Urinary Tract Infections/diagnosis , Urinary Tract Infections/microbiology , Bacteriological Techniques/instrumentation , Candida/classification , Chromogenic Compounds , Colony Count, Microbial , Culture Media , Gram-Negative Bacteria/classification , Gram-Positive Bacteria/classification , Humans , Reproducibility of Results , Sensitivity and Specificity , UrineABSTRACT
The new BDProbeTec ET Mycobacterium tuberculosis Complex Direct Detection Assay (DTB) was compared with the enhanced M. tuberculosis Amplified Direct Test (AMTDII). The system is an automated walkaway system characterized by simultaneous DNA amplification (strand displacement amplification) and real-time fluorometric detection. It also contains an internal amplification control (IAC) designed to identify inhibition from the processed samples. The AMTDII assay amplifies rRNA by transcription-mediated amplification; it uses hybridization with a chemoluminescent probe as a detection system and is entirely manual. A total of 515 N-acetyl-L-cysteine-sodium hydroxide-decontaminated respiratory (n = 331) and extrapulmonary (n = 184) sediments (from 402 patients) were tested in parallel by both assays. The results were compared with those of acid-fast staining and culture (solid plus liquid media), setting the combination of culture and clinical diagnosis as the "gold standard." Culture results from the tested specimens were as follows: 121 Mycobacterium tuberculosis complex (MTB) (98 smear-positive), 46 nontuberculous mycobacteria (38 smear-positive), and 338 culture-negative results. After resolution of the discrepant results, the percent sensitivity, percent specificity, and positive and negative likelihood ratios for AMTDII were 88%, 99.2%, 110, and 0.11 for respiratory specimens and 74.3%, 100%, 740, and 0.26 for extrapulmonary specimens, respectively. The corresponding values for DTB were 94.5%, 99.6%, 235, and 0.05 for respiratory specimens and 92.3%, 100%, 920, and 0.07 for extrapulmonary specimens, respectively. The cumulative difference for all tuberculosis-positive extrapulmonary specimens was significant (P = 0.03). The overall inhibition rate for DTB was 5% (26 specimens). We conclude that both amplification assays proved to be rapid and specific for the detection of MTB in clinical samples and particularly feasible for a routine laboratory work flow. DTB combines a labor-intensive specimen preparation procedure with a completely automated amplification and detection. Finally, differences between AMTDII and DTB sensitivities were associated with the presence of inhibitory samples that the former assay, lacking IAC, could not detect.
Subject(s)
Mycobacterium tuberculosis/isolation & purification , Nucleic Acid Amplification Techniques/methods , Respiratory System/microbiology , Tuberculosis, Pulmonary/microbiology , Tuberculosis/microbiology , Humans , Molecular Probe Techniques , Mycobacterium tuberculosis/genetics , Predictive Value of Tests , Reagent Kits, Diagnostic , Reproducibility of Results , Sensitivity and SpecificityABSTRACT
We evaluated the BDProbeTec ET system (Becton Dickinson, Sparks, Md.), a strand displacement amplification-based technique, for direct detection of Mycobacterium tuberculosis in 867 clinical samples. Of 294 extrapulmonary specimens, 52 had positive results by both BDProbeTec ET and culture and 209 had negative results by both methods; sensitivity and specificity were 76.5 and 95.9%, respectively. After resolution of discrepancies, the sensitivity rose to 77.8%.