Donor-derived mold infections in lung transplant recipients: The importance of active surveillance.

Unexpected donor-derived fungal infections represent a rare but potentially fatal complication in lung transplant (Tx) recipients. Timely communication of the results of donor cultures and prompt treatment of recipients are crucial to mitigate the consequences of donor-derived transmissions. In this prospective cohort study, all consecutive patients who underwent lung transplantation from 2015 to 2022 were included. In December 2015, a Local Active Surveillance System has been implemented to provide biovigilance of donor culture results and optimize recipients' management. The aim of this study is to investigate the incidence of unexpected, mold-positive cultures among lung donors and the rate of transmission to recipients. Furthermore, management strategies and outcome of recipients with mold transmission are described. In case of isolation of the same mold in donor and recipient cultures, when possible, transmission was confirmed by dendrogram analysis. During the study period, 82 lung Tx were performed from 80 donors. The prevalence of donors with "unexpected" mold isolation from the respiratory tract was 3.75% (3/80). Isolated molds were Aspergillus niger, Rhizopus oryzae, and Aspergillus flavus. Transmissions occurred in all the three cases (100%) with a mean time of 5 days from lung Tx but none of the recipients developed invasive mold disease. Our Local Active Surveillance System allowed prompt recognition of lung donors unexpected mold colonization. Even though transmission occurred, introduction of early targeted antifungal therapy prevented potential catastrophic consequence of mold donor-derived infection in the immediate post-Tx period.

Phaeohyphomycosis in Solid Organ Transplant Recipients: A Case Series and Narrative Review of the Literature.

Phaeohyphomycosis comprises a variety of infections caused by pigmented fungi. Solid organ transplant (SOT) recipients are particularly at risk of invasive infections due to their prolonged immunosuppression. Here, we describe three cases of phaeohyphomycosis in SOT recipients who were successfully treated with surgical excision and/or antifungal therapy. We additionally carried out a narrative review of the literature on phaeohyphomycosis in 94 SOT recipients from 66 published studies describing 40 different species of fungi. The most reported fungus was Alternaria (21%). The median time from transplant to diagnosis was 18 months (IQR 8.25-48), and kidney transplants were the most reported. Antifungal regimens were not homogeneous, though there was a prevalence of itraconazole- and voriconazole-based treatments. Clinical outcomes included recovery in 81% and death in 5% of infected SOT recipients. Susceptibility testing was done in 26.6% of the cases, with heterogeneous results due to the variety of species isolated. While the wide diversity of dematiaceous fungi and their host range make it difficult to offer a uniform approach for phaeohyphomycosis, an early diagnosis and therapy are critical in preventing the dissemination of disease in the immunocompromised host.

Erratum: Complete intra-laboratory validation of a LAL assay for bacterial endotoxin determination in EBV-specific cytotoxic T lymphocytes.

[This corrects the article DOI: 10.1016/j.omtm.2021.05.002.].

Complete intra-laboratory validation of a LAL assay for bacterial endotoxin determination in EBV-specific cytotoxic T lymphocytes.

Endotoxin content is a critical factor that affects the safety of biological pharmaceutical products. International pharmacopoeias describe several reference methods to determine endotoxin levels in advanced therapy medicinal product (ATMP) preparations. Administration of ATMPs must be done as rapidly as possible to ensure complete viability and potency of the cellular product. To evaluate the endotoxin content in the shortest time possible, we chose to validate an alternative method based on the use of the Charles River Portable Testing System (PTS) and FDA-approved cartridges, compliant with the requirements of the European Pharmacopoeia and providing results in <20 min. Here, we describe a unique and complete validation approach for instrument, personnel, and analytical method for assessment of endotoxins in ATMP matrices. The PTS system provides high sensitivity and fast quantitative results and uses less raw material and accessories compared with compendial methods. It is also less time consuming and less prone to operator variability. Our validation approach is suitable for a validated laboratory with trained personnel capable of conducting the ATMP release tests, and with very low intra-laboratory variability, and meets the criteria required for an alternative approach to endotoxin detection for in-process and product-release testing of ATMPs.

Microbiological Surveillance of Endoscopes in a Southern Italian Transplantation Hospital: A Retrospective Study from 2016 to 2019.

Endoscopes are medical instruments that are used routinely in health structures. Due to their invasive nature and contact with many patients, they may cause hospital-acquired infections if not disinfected correctly. To ensure a high-level disinfection procedure or reprocessing, since the methods currently adopted in our institute are adequate, we evaluated retrospectively the presence of microorganisms in our endoscopes after reprocessing. Microbiological surveillance was performed from January 2016 to December 2019 in the instruments in use in our endoscopic room after reprocessing. In total, 35 endoscopes (3 duodenoscopes, 3 echoendoscopes, 12 bronchoscopes, 5 colonoscopes, and 12 gastroscopes) were evaluated for the presence of microorganisms, including multidrug-resistant pathogens and indicator microorganisms (IMOs). Our procedures were in agreement with an internal protocol based on Italian, international, and the Center for Disease Control and Prevention (CDC) recommendations. Of a total of 811 samples, 799 (98.5%) complied with the regulatory guidelines, while 9 (1.1%) were positive for IMOs, and 3 (0.4%) displayed more than 10 colony-forming units (CFU) of environmental and commensal pathogens. Our results show that the internal reprocessing protocol is very efficient, leading to a very low number of observed contaminations, and it could be easily implemented by other health facilities that face a huge number of hospital-acquired infections due to incorrectly disinfected endoscopes.

Efficacy of Three Commercial Disinfectants in Reducing Microbial Surfaces' Contaminations of Pharmaceuticals Hospital Facilities.

To evaluate and validate the efficacy of disinfectants used in our cleaning procedure, in order to reduce pharmaceutical hospital surfaces' contaminations, we tested the action of three commercial disinfectants on small representative samples of the surfaces present in our hospital cleanrooms. These samples (or coupons) were contaminated with selected microorganisms for the validation of the disinfectants. The coupons were sampled before and after disinfection and the microbial load was assessed to calculate the Log10 reduction index. Subsequently, we developed and validated a disinfection procedure on real surfaces inside the cleanrooms intentionally contaminated with microorganisms, using approximately 107-108 total colony forming units per coupon. Our results showed a bactericidal, fungicidal, and sporicidal efficacy coherent to the acceptance criteria suggested by United States Pharmacopeia 35 <1072>. The correct implementation of our cleaning and disinfection procedure, respecting stipulated concentrations and contact times, led to a reduction of at least 6 Log10 for all microorganisms used. The proposed disinfection procedure reduced the pharmaceutical hospital surfaces' contaminations, limited the propagation of microorganisms in points adjacent to the disinfected area, and ensured high disinfection and safety levels for operators, patients, and treated surfaces.

Strategy and validation of a consistent and reproducible nucleic acid technique for mycoplasma detection in advanced therapy medicinal products.

Advanced therapy medicinal products (ATMP) are required to maintain their quality and safety throughout the production cycle, and they must be free of microbial contaminations. Among them, mycoplasma contaminations are difficult to detect and undesirable in ATMP, especially for immunosuppressed patients. Mycoplasma detection tests suggested by European Pharmacopoeia are the "culture method" and "indicator cell culture method" which, despite their effectiveness, are time consuming and laborious. Alternative methods are accepted, provided they are adequate and their results are comparable with those of the standard methods. To validate a novel in-house method, we performed and optimized, a real time PCR protocol, using a commercial kit and an automatic extraction system, in which we tested different volumes of matrix, maximizing the detection sensitivity. The results were compared with those obtained with the gold standard methods. From a volume of 10 ml, we were able to recognize all the mycoplasmas specified by the European Pharmacopoeia, defined as genomic copies per colony forming unit ratio (GC/CFU). Our strategy allows to achieve faster and reproducible results when compared with conventional methods and meets the sensitivity and robustness criteria required for an alternative approach to mycoplasmas detection for in-process and product-release testing of ATMP.

Kartagener's syndrome: review of a case series.

BACKGROUND: Kartagener Syndrome (KS) is a rare autosomal recessive genetic disorder, resulting in a group of clinical manifestations, including bronchiectasis, chronic pansinusitis and situs inversus. METHODS: We hereby reviewed eight cases of this rare entity selected from patients attending our outpatients Respiratory Unit since 2006. Samples of respiratory epithelium were obtained with the method of nasal brushing and sent to a specialized center in order to be studied with electron microscopy. At least 50 cross sections of different cilia from different cells were observed in each specimen to study the axonemal structure. Electron micrographs were taken at a magnification of X 50,000 to determine the orientation of the cilia and at a magnification of X 110,000 to study the axonemal pattern. The incidence of abnormal cilia was expressed as a percentage. RESULTS: We observed different ultrastructural defects in our KS patients, including absence of outer dynein arms, absence of outer and inner dynein arms, and absence of the central pair with transposition of a peripheral doublet into the central position. Patient's follow up lasted till 2014, however two patients with more severe clinical behavior died before. CONCLUSIONS: This is a review of a case series, yet our data has shown that nasal brushing with ultrastructural pathological differentiation may be useful to identify patients with high risk and to develop more complex clinical presentations.

Retention of quinolones on human serum albumin and alpha1-acid glycoprotein HPLC columns: relationships with different scales of lipophilicity.

The retention of 10 quinolone antibacterial agents on HPLC stationary phases supporting human serum albumin (HSA) or alpha(1)-acid glycoprotein (AGP) was investigated. Among ofloxacine and flumequine, the two chiral compounds in the selected set, only the latter showed a split chromatographic peak and only on HSA but not on AGP, indicating that enantioselective specific sites play only a minor role in the retention. The retention of quinolones, which included four acidic and six zwitterionic congeners, was correlated with various lipophilicity scales: (i) theoretically calculated values, clogP, (ii) values measured at pH 7.4 by the shake-flask method, logD(7.4), and (iii) values extrapolated by retention data measured by ion-pair reversed-phase high performance liquid chromatography (RP-HPLC). We assumed that the latter values, logP(i.p.), were close to the lipophilicity of the neutral forms, logP(N), for both acidic and zwitterionic congeners. No relationship was found between retention on serum proteins and clogP values, whereas a reasonable relationship was found with logD(7.4) values, but only when the two subclasses, acidic and zwitterionic congeners, were considered separately. The relationship between retention data on serum proteins and logP(i.p.) values indicated that the affinity for serum proteins depends on the lipophilicity of the neutral forms only for logP values up to 1.5. Above this value, protein retention does not further increase, becoming almost constant. Based on both the observations above reported and the small values of the slopes of regression equations, we conclude that the interaction of the more lipophilic quinolones, mainly the zwitterions, with serum proteins is not governed uniquely by lipophilicity but also by other mechanisms, probably of electrostatic nature.

Can protonated beta-blockers interact with biomembranes stronger than neutral isolipophilic compounds? A chromatographic study on three different phospholipid stationary phases (IAM-HPLC).

The chromatographic capacity factors (k') of 10 beta-adrenoceptor antagonists ("beta-blockers") were measured on three different immobilized artificial membrane-phosphatidylcholine (IAM-PC) HPLC stationary phases, namely IAM-PC-MG, IAM-PC-DD2, and IAM-PC-DD. The two former phases are made of phosphatidylcholine as found in biomembranes and differ each other in end-capping of free propylamino residues whereas the latter is made of single-chain phosphatidylcholine analogues. On IAM-PC-DD2 we found that structurally unrelated neutral compounds give a single correlation between logk' values and the respective octanol/water partition coefficients (logP), as previously observed on IAM-PC-MG phase. This was not observed on the IAM-PC-DD phase. IAM chromatography was performed at a pH of the aqueous eluent (7.0) close to the physiological pH 7.4. Retention on all IAM phases showed a biphasic pattern, being proportional to logP(N) (lipophilicity of neutral forms) for more lipophilic congeners (logP(N)>1.90), and quite constant for the others. The comparison of beta-blocker retention with that of neutral compounds on IAM-PC-MG and IAM-PC-DD2 suggests that they can interact with phospholipids as strongly or more strongly than neutral isolipophilic compounds, despite their being more than 98% in their ionized form. Therefore, we hypothesize that electrostatic interactions play a pivotal role in the interactions between beta-blockers and membrane phospholipids.