https://www.srcv.ro/en/ghiduri/Preventia%20trombembolismului%20la%20pacientii%20nonortopedici%20-%20Ghidurile%20ACCP%202012.pdf

Prevention of VTE in Nonorthopedic Surgical Patients : Antithrombotic Therapy and Prevention of Thrombosis, 9th ed: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines

Invasive aspergillosis occurs in a wide range of clinical scenarios, is protean in its manifestations, and is still associated with an unacceptably high mortality rate. Early diagnosis is critical to a favourable outcome, but is difficult to achieve with current methods. Deep tissue diagnostic specimens are often difficult to obtain from critically ill patients. Newer antifungal agents exhibit differential mould activity, thus increasing the importance of establishing a specific diagnosis of invasive aspergillosis. For these reasons, a range of alternate diagnostic strategies have been investigated. Most investigative efforts have focused on molecular and serological diagnostic techniques. The detection of metabolites produced by Aspergillus spp and a range of aspergillus-specific antibodies represent additional, but relatively underused, diagnostic avenues. The detection of galactomannan has been incorporated into diagnostic criteria for invasive aspergillosis, reflecting an increased understanding of the performance, utility, and limitations of this technique. Measurement of (1,3)-beta-D glucan in blood may be useful as a preliminary screening tool for invasive aspergillosis, despite the fact that this antigen can be detected in a number of other fungi. There have been extensive efforts directed toward the detection of Aspergillus spp DNA, but a lack of technical standardisation and relatively poor understanding of DNA release and kinetics continues to hamper the broad applicability of this technique. This review considers the application, utility, and limitations of the currently available and investigational diagnostic modalities for invasive aspergillosis.

/pdf/laboratory-diagnosis-of-invasive-aspergillosis-435zvpzyx7.pdf

Laboratory diagnosis of invasive aspergillosis

Given the rise in the incidence of invasive fungal infections (IFIs) and the expanding spectrum of fungal pathogens, early and accurate identification of the causative pathogen is essential. We developed a panfungal PCR assay that targets the internal transcribed spacer 1 (ITS1) region of the ribosomal DNA gene cluster to detect fungal DNA in fresh and formalin-fixed, paraffin-embedded (PE) tissue specimens from patients with culture-proven (n = 38) or solely histologically proven (n = 24) IFIs. PCR products were sequenced and compared with sequences in the GenBank database to identify the causal pathogen. The molecular identification was correlated with results from histological examination and culture. The assay successfully detected and identified the fungal pathogen in 93.6% and 64.3% of culture-proven and solely histologically proven cases of IFI, respectively. A diverse range of fungal genera were identified, including species of Candida, Cryptococcus, Trichosporon, Aspergillus, Fusarium, Scedosporium, Exophiala, Exserohilum, Apophysomyces, Actinomucor, and Rhizopus. For five specimens, molecular analysis identified a pathogen closely related to that identified by culture. All PCR-negative specimens (n = 10) were PE tissues in which fungal hyphae were visualized. The results support the use of the panfungal PCR assay in combination with conventional laboratory tests for accurate identification of fungi in tissue specimens.

Development and Clinical Application of a Panfungal PCR Assay To Detect and Identify Fungal DNA in Tissue Specimens

Current knowledge on the opportunist Scedosporium apiospermum (teleomorph: Pseudallescheria boydii), generated over a period of more than 120 years, is reviewed. The natural environmental habitat of the fungus is unknown; nutrient-rich, brackish waters like river estuaria have been suggested. The fungus is strongly promoted by agricultural and particularly by industrial pollution.

/pdf/scedosporium-apiospermum-changing-clinical-spectrum-of-a-c27xmr6vwn.pdf

Scedosporium apiospermum: changing clinical spectrum of a therapy-refractory opportunist.

Molecular approaches are now being developed to provide a more rapid and objective identification of fungi compared to traditional phenotypic methods. Ribosomal targets, especially the large-subunit RNA gene (D1-D2 region) and internal transcribed spacers 1 and 2 (ITS1 and ITS2 regions), have shown particular promise for the molecular identification of some fungi. We therefore conducted an assessment of these regions for the identification of 13 medically important Aspergillus species: Aspergillus candidus, Aspergillus (Eurotium) chevalieri, Aspergillus (Fennellia) flavipes, Aspergillus flavus, Aspergillus fumigatus, Aspergillus granulosus, Aspergillus (Emericella) nidulans, Aspergillus niger, Aspergillus restrictus, Aspergillus sydowii, Aspergillus terreus, Aspergillus ustus, and Aspergillus versicolor. The length of ribosomal regions could not be reliably used to differentiate among all Aspergillus species examined. DNA alignment and pairwise nucleotide comparisons demonstrated 91.9 to 99.6% interspecies sequence identities in the D1-D2 region, 57.4 to 98.1% in the ITS1 region, and 75.6 to 98.3% in the ITS2 region. Comparative analysis using GenBank reference data showed that 10 of the 13 species examined exhibited a ≤1-nucleotide divergence in the D1-D2 region from closely related but different species. In contrast, only 5 of the species examined exhibited a ≤1-nucleotide divergence from sibling species in their ITS1 or ITS2 sequences. Although the GenBank database currently lacks ITS sequence entries for some species, and major improvement in the quality and accuracy of GenBank entries is needed, current identification of medically important Aspergillus species using GenBank reference data seems more reliable using ITS query sequences than D1-D2 sequences, especially for the identification of closely related species.

https://jcm.asm.org/content/jcm/43/5/2092.full.pdf

Assessment of Ribosomal Large-Subunit D1-D2, Internal Transcribed Spacer 1, and Internal Transcribed Spacer 2 Regions as Targets for Molecular Identification of Medically Important Aspergillus Species

Background Deep venous thrombosis (DVT) and pulmonary embolism (PE) may be significant complications following spinal surgery. The incidence rate ranges from 0.5% to 2.5% in patients with symptomat...

Thromboembolic complications after spinal surgery in trauma patients

The aim of this study was to find a reliable method for the detection and identification of fungi in fungus balls of the maxillary sinus and to evaluate the spectrum of fungi in these samples. One hundred twelve samples were obtained from patients with histologically proven fungal infections; 81 samples were paraffin-embedded tissue sections of the maxillary sinus. In 31 cases, sinus contents without paraffin embedding were sent for investigation. PCR amplification with universal fungal primers for 28S ribosomal DNA and amplicon identification by hybridization with species-specific probes for Aspergillus fumigatus, Aspergillus flavus, Aspergillus niger, Aspergillus terreus, Aspergillus glaucus, Pseudallescheria boydii, Candida albicans, and Candida glabrata were performed for all samples. Furthermore, PCR products were sequenced. Fresh samples were also cultivated. Fungal DNA was detected in all of the fresh samples but only in 71 paraffin-embedded tissue samples (87.7%). Sequence analysis was the most sensitive technique, as results could be obtained for 28 (90.3%) fresh samples by this method in comparison to 24 (77.4%) samples by hybridization and 16 (51.6%) samples by culture. However, sequence analysis delivered a result for only 36 (50.7%) of the paraffin-embedded specimens. Hybridization showed reliable results for A. fumigatus, which proved to be the most common agent in fungus balls of the maxillary sinus. Other Aspergillus species and other genera were rarely found.

Detection and Identification of Fungi from Fungus Balls of the Maxillary Sinus by Molecular Techniques

Invasive fungal infection may afflict people with trauma in two ways: either by entry into tissue via penetrating trauma or by haematogenous spread in critically ill people with polytrauma. Penetrating injury allows the advance of ubiquitously present fungi into the human body. Miniscule foreign material fosters the establishment and growth of fungi within the traumatically changed tissue. The seriousness of the infection depends upon the type of injury, the body area and the person's general condition. Usually, the infection is confined to the cutis and subcutis; the fascia, muscles and bones are rarely affected. In the presence of immunocompromise, however, the fungus may spread rapidly and cause systemic disease. The following overview will focus on fungal infection associated with open wounds and fractures, particularly eye injury and with near-drowning, tropical mycetoma and nosocomial conditions. Post-traumatic invasive fungal infections are rare, but the surgeon should be alert to this possibility in cases with chronic inflammation and deferred healing of injuries, with or without systemic inflammatory response.

Invasive mycoses following trauma

Das eingesprengte Fremdmaterial bei einer offenen Verletzung begunstigt die Inokulation ubiquitar vorhandener Pilze, die in traumatisch verandertem Gewebe sich festsetzen und vermehren konnen. Akut verlaufende Pilzinfektionen infolge eines perforierenden Traumas sind sehr selten. Die Schwere der Infektion ist von der Art der Verletzung (penetrierend, Fremdkorper), der Korperregion und vom Allgemeinzustand des Patienten abhangig. Bei immunkompetenten Patienten bleibt die Infektion lokal auf Kutis und Subkutis beschrankt, seltener sind Faszien, Muskeln und Knochen betroffen. Diese Ubersichtsarbeit befasst sich mit Pilzinfektionen nach offenen Verletzungen. Die invasiven Pilzinfektionen werden in folgende Gruppen eingeteilt: invasive Mykosen durch Inokulation als direkte Verletzungsfolge, nach Augenverletzungen, als tropische Pilzerkrankung (Myzetom, Madurafuss), nach (Beinahe-)Ertrinken und invasive Mykosen als nosokomiale Infektion bei Langzeitaufenthalt auf Intensivstationen nach schwerem Polytrauma.

Alexandra Obradovic

Papers

Thromboembolic complications after spinal surgery in trauma patients

Detection and Identification of Fungi from Fungus Balls of the Maxillary Sinus by Molecular Techniques

Invasive mycoses following trauma

Invasive mycoses and trauma