Targeting essential pathways in trypanosomatids gives insights into protozoan mechanisms of cell death

Apoptosis is a normal component of the development and health of multicellular organisms. However, apoptosis is now considered a prerogative of unicellular organisms, including the trypanosomatids of the genera Trypanosoma spp. and Leishmania spp., causative agents of some of the most important neglected human diseases. Trypanosomatids show typical hallmarks of apoptosis, although they lack some of the key molecules contributing to this process in metazoans, like caspase genes, Bcl-2 family genes and the TNF-related family of receptors. Despite the lack of these molecules, trypanosomatids appear to have the basic machinery to commit suicide. The components of the apoptotic execution machinery of these parasites are slowly coming into light, by targeting essential processes and pathways with different apoptogenic agents and inhibitors. This review will be confined to the events known to drive trypanosomatid parasites to apoptosis

Expansion of KPC-producing klebsiella pneumoniae with various mgrB-mutations giving rise to colistin-resistance:the role of ISL3 on plasmids

BACKGROUND: mcr-1 has been reported as first plasmid gene to confer colistin-resistance. In KPC-producing Klebsiella pneumoniae (KPC-KP) isolates, however, colistin resistance is rapidly emerging through other mechanisms. This is frequently the result of disruption of mgrB gene by insertion sequences, for instance ISL3. The aim of this study is to investigate the expansion of mgrB-mutated KPC-KP. In addition, we identify localization and targets of ISL3 sequences within the core and accessory genome of common KPC-KP lineages. METHODS: Twenty-nine clinical isolates of K. pneumoniae collected from Italian patients were randomly selected. Whole genome sequences (WGS) were analysed for resistance genes, plasmids, and insertion sequences. Additionally, 27 colistin-resistant KPC-KP isolates from a previous study from Crete was assessed. FINDINGS: We observed clonal expansion of KPC-KP isolates with various mutations in mgrB among all lineages. In two Italian MLST ST512 isolates, and eight Greek ST258 isolates an identical copy of the ISL3 was inserted in mgrB nucleotide position 133. ISL3s, transposable restriction modification systems of 8154 nucleotides, were positioned on pKpQIL carrying isolates and may transpose into the chromosome. In 4 isolates, chromosomal integration of ISL3 in diverse inner-membrane proteins other than mgrB was identified. INTERPRETATION: Colistin resistance is most often explained by clonal expansion of isolates with mutated mgrB. pKpQIL-like plasmids, which are omnipresent in KPC-KP, carry insertion sequences such as ISL3 that have mgrB as a target hotspot for transposition. Transposition of insertion sequences from plasmids and subsequent clonal expansion may contribute to the emerging colistin resistance in KPC-KP

Synergistic combination of alkylphosphocholines with peptaibols in targeting Leishmania infantum in vitro

Anti-leishmanial treatment increasingly encounters therapeutic limitations due to drug toxicity and development of resistance. The effort for new therapeutic strategies led us to work on combinations of chemically different compounds that could yield enhanced leishmanicidal effect. Peptaibols are a special type of antimicrobial peptides that are able to form ion channels in cell membranes and potentially affect cell viability. We assayed the antileishmanial activity of two well studied helical peptaibols, the 16-residue antiamoebin and the 20-residue alamethicin-analogue suzukacillin, and we evaluated the biological effect of their combination with the alkylphosphocholine miltefosine and its synthetic analogue TC52. The peptaibols tested exhibited only moderate antileishmanial activity, however their combination with miltefosine had a super-additive effect against the intracellular parasite (combination index 0.83 and 0.43 for antiamoebin and suzukacillin respectively). Drug combinations altered the redox stage of promastigotes, rapidly dissipated mitochondrial membrane potential and induced concatenation of mitochondrial network promoting spheroidal morphology. These results evidenced a potent and specific antileishmanial effect of the peptaibols/miltefosine combinations, achieved with significantly lower concentrations of the compounds compared to monotherapy. Furthermore, they revealed the importance of exploring novel classes of bioactive compounds such as peptaibols and demonstrated for the first time that they can act in synergy with currently used antileishmanial drugs to improve the therapeutic outcome. Keywords: Leishmaniasis therapy, Miltefosine, Peptaibol antibiotics, Drug synergy, Mitochondrial membrane potential, Reactive oxygen specie

Clinical and microbiological characteristics of nocardiosis: A 5-year single-center study in Crete, Greece

Nocardiosis is a rare disease affecting both immunocompromised and immunocompetent hosts, presented in various clinical forms ranging from localized to disseminated infection. Aim of the present study was to investigate the clinical and microbiological characteristics of nocardiosis, antimicrobial resistance profiles, treatment, and outcomes of Nocardia infection over the last 5 years at our institution. The medical records and microbiological data of patients affected by nocardiosis and treated at the university hospital of Heraklion, Crete, Greece, between 2018 and 2022, were retrospectively analyzed. The isolates were identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and through sequencing of 16S rRNA. Antimicrobial susceptibility for 17 agents was determined by E-test and results were interpreted according to CLSI guidelines. Among the 28 Nocardia isolates, eight species were identified, with Nocardia brasiliensis being the most prevalent (32.1%), followed by Nocardia otitidiscaviarum (25%), and Nocardia farcinica (14.3%). Skin and soft tissue infections were the most common presentations, noted in 13 (50%) patients, followed by pulmonary infection presented in 10 (38.5%) patients. Fifteen patients (57.7%) had at least one underlying disease, and 11 (42.3%) were on immunosuppressive or long-term corticosteroid treatment. Susceptibility rates of linezolid, tigecycline, amikacin, trimethoprim-sulfamethoxazole, moxifloxacin, and imipenem were 100, 100, 96.4, 92.9, 82.1, and 42.9%, respectively. The 26 patients in this study were treated with various antibiotics. Mortality rate was 3.8%, and the patient who died had disseminated infection. Since epidemiology and antimicrobial susceptibility are evolving, continuous surveillance is mandatory in order to initiate appropriate treatment in a timely manner

Nocardia elegans primary iliopsoas abscess: A case report and literature review

Nocardia species are rare causative agents of psoas abscess, more frequently occurring as part of disseminated infection. Only sporadic cases have been reported so far, with Nocardia asteroides and Nocardia farcinica being the most common causative agents. Nocardia elegans is an opportunistic pathogen, accounting for only 0.3-0.6% of infections caused by Nocardia species, usually affecting the respiratory tract. In this study, a previously healthy 74-year-old man was admitted to the University Hospital of Heraklion with fever and intense pain radiating from the lumbar region to the groin and the left thigh, increasing with movement. Imaging findings revealed a large abscess in the left iliopsoas. Blood and pus aspirate cultures yielded a pure culture of Nocardia that was identified by 16S rRNA sequence as N. elegans. The patient was successfully treated with drainage of the abscess along with administration of ceftriaxone, linezolid and trimethoprim-sulfamethoxazole. To our knowledge, this is the first report of iliopsoas abscess caused by N. elegans. Early, accurate diagnosis and timely treatment with drainage of the abscess and long-term administration of antimicrobial agents optimize the outcome

Bicuspid aortic valve endocarditis caused by Gemella sanguinis: Case report and literature review

Gemella species are catalase-negative, facultative anaerobic, Gram-positive cocci, which are part of the human oral microbiome and may occasionally cause systemic infections. Infective endocarditis (IE) has been reported as the most common infection caused by Gemella species. We report the first case of IE due to Gemella sanguinis in Greece, in a patient with bicuspid aortic valve and review the available literature. The patient was successfully treated with antibiotics and aortic valve replacement. Keywords: Gemella sanguinis, Infective endocarditis, Bicuspid aortic valve, Aortic regurgitatio