EVALUATION OF RPMI-PY MEDIUM FOR TRYPANOSOMA CRUZI AND DIFFERENT LEISHMANIA SPECIES

Trypanosoma spp. and Leishmania spp. are causal agents of a number of parasitic diseases. Culture media can be divided into 3 main categories: semisolid, biphasic, and liquid. While biphasic and semisolid culture media need blood, an important factor for the reproduction of parasites, most liquid media require fetal calf serum or erythrocyte lysate. A culture media RPMI-PY demonstrated a good performance in terms of time and parasitic load of L. infantum compared to other culture media. The aim of the work was to evaluate the performance of RPMI-PY medium in different Leishmania species and also to evaluate in T. cruzi culture. RPMI-PY is likely to be valuable additions to laboratory practice in light of the relatively simple recipes, general availability of the components, and in terms of suitability because rabbit breeding is not necessary and the costs are lowered and can be used for all Leishmania species and to cultivate T. cruzi

STILBENE ST18 AND TERFENYL TR4: IN VITRO ACTIVITY AGAINST TRYPANOSOMA CRUZI

Chagas disease is caused by the parasite Trypanosoma cruzi, which is transmitted to animals and people by insect vectors that are found only in the Americas. Chagas disease and Leishmaniasis are life-threatening illnesses caused by the protozoan parasites Leishmania spp. and Trypanosoma cruzi, respectively. They are known as “neglected diseases” due to the lack of effective drug treatments and the scarcity of research work devoted to them. Therefore, the development of novel and effective drugs is an important and urgent need. Natural products are an important source of bioactive molecules for the development of new drugs. Recently, studies showed an interesting cytotoxic action of Stilbene ST18 and the Terphenyl TR4 compounds in Leishmania. In this study, we evaluated the in vitro trypanocidal activity of ST18 and TR4 and Nifurtimox, drug used for the treatment of Chagas disease. In addition, we evaluated the compounds action in infected macrophages with Trypanosoma cruzi. Results showed that three compounds exhibited significant activity against Trypanosoma compared to Nifurtimox. ST18 and TR4 compounds inhibited Trypanosoma growth with IC50 values of 4.5 and 32 μM, respectively. The treatment of infected macrophages with trypanosomes with the IC90 compounds showed a reduction of infection compared to control: ST18 reduced the infected cells to 52 %, TR4 reduced the infected cells to 63%. In conclusion, these news compounds could be considered as promising lead drugs for the development of new therapies for the treatment of Chagas disease

Role of alpha-tocopherol in counteracting DNA damage induced by Ochratoxin A in primary porcine fibroblasts

Ochratoxin A is a mycotoxin responsible for disease states in both humans and animals. OTA mechanisms of action are numerous, including lipid peroxidation. Oxidative damage results in the modification of macromolecules (i.e. DNA), cell death and tissue injure. Several strategies, such as the use of antioxidants, have been used to reduce OTA cytotoxicity. The aim of this study was to evaluate the role of alpha-tocopherol in counteracting DNA damage induced by OTA in cell cultures. Primary porcine fibroblasts, isolated from embryo and from ear, were incubated for 24h with several concentrations of OTA in order to detect DNA fragmentation. OTA produced DNA fragmentation in a concentration dependent manner in both primary cell cultures. The pre-treatment with alpha-tocopherol caused the reduction of DNA fragmentation in both primary cell cultures, after 24h of incubation with OTA. In particular, when OTA was added at 10 µg/ml in embryo fibroblasts, alpha-tocopherol at the concentrations of 1 nM was significantly (P<0.05) able to reduce DNA fragmentation by 16%. In ear fibroblast cultures, alpha-tocopherol at the 1nM concentration was significantly (P<0.05) able to reduce DNA fragmentation by 15.23% in the presence of 5 µg/ml of OTA

Overview of DISCOVER22 experiment in the framework of INFN-LNGS Cosmic Silence activity: challenges and improvements in underground radiobiology

One of the most intriguing and still pending questions in radiobiology is to understand whether and how natural environmental background radiation has shaped Life over millions of years of evolution on Earth. Deep Underground Laboratories (DULs) represent the ideal below-background exposure facilities where to address such a question. Among the few worldwide DULs, INFN-Laboratorio Nazionale del Gran Sasso (LNGS) is one of the largest in terms of size and infrastructure. Designed and built to host neutrino and dark matter experiments, since the 1990 s the LNGS has been one of the first DULs to systematically host radiobiology experiments. Here we present the DISCOVER22 (DNA Damage and Immune System Cooperation in VEry low Radiation environment 2022) experiment recently started at LNGS. DISCOVER22 aims at investigating how the low radiation background modulates the Immune System (IS) response in in vitro and in vivo models. Underground radiobiology experiments are particularly complex and tricky to design and perform. In these studies, the accurate characterization of exposure scenarios is mandatory, but a challenging aspect is to understand how the very few ionizing tracks in the ultra-Low Radiation Environment (LRE) interact with the living matter in space and time in order to trigger different biological responses. In this Perspective, we describe these challenges and how we address them through a microdosimetric and a radiobiological approaches. We aim at linking physical microdosimetric measurements and the corresponding biological radiation responses by using radiation biophysical models that could shed light on many as yet unresolved questions

Operative management of acute abdomen after bariatric surgery in the emergency setting: the OBA guidelines

Background: Patients presenting with acute abdominal pain that occurs after months or years following bariatric surgery may present for assessment and management in the local emergency units. Due to the large variety of surgical bariatric techniques, emergency surgeons have to be aware of the main functional outcomes and long-term surgical complications following the most performed bariatric surgical procedures. The purpose of these evidence-based guidelines is to present a consensus position from members of the WSES in collaboration with IFSO bariatric experienced surgeons, on the management of acute abdomen after bariatric surgery focusing on long-term complications in patients who have undergone laparoscopic sleeve gastrectomy and laparoscopic Roux-en-Y gastric bypass. Method: A working group of experienced general, acute care, and bariatric surgeons was created to carry out a systematic review of the literature following the Preferred Reporting Items for Systematic Review and Meta-analysis Protocols (PRISMA-P) and to answer the PICO questions formulated after the Operative management in bariatric acute abdomen survey. The literature search was limited to late/long-term complications following laparoscopic sleeve gastrectomy and laparoscopic Roux-en-Y gastric bypass. Conclusions: The acute abdomen after bariatric surgery is a common cause of admission in emergency departments. Knowledge of the most common late/long-term complications (&gt; 4 weeks after surgical procedure) following sleeve gastrectomy and Roux-en-Y gastric bypass and their anatomy leads to a focused management in the emergency setting with good outcomes and decreased morbidity and mortality rates. A close collaboration between emergency surgeons, radiologists, endoscopists, and anesthesiologists is mandatory in the management of this group of patients in the emergency setting

Correction to: Two years later: Is the SARS-CoV-2 pandemic still having an impact on emergency surgery? An international cross-sectional survey among WSES members

Background: The SARS-CoV-2 pandemic is still ongoing and a major challenge for health care services worldwide. In the first WSES COVID-19 emergency surgery survey, a strong negative impact on emergency surgery (ES) had been described already early in the pandemic situation. However, the knowledge is limited about current effects of the pandemic on patient flow through emergency rooms, daily routine and decision making in ES as well as their changes over time during the last two pandemic years. This second WSES COVID-19 emergency surgery survey investigates the impact of the SARS-CoV-2 pandemic on ES during the course of the pandemic. Methods: A web survey had been distributed to medical specialists in ES during a four-week period from January 2022, investigating the impact of the pandemic on patients and septic diseases both requiring ES, structural problems due to the pandemic and time-to-intervention in ES routine. Results: 367 collaborators from 59 countries responded to the survey. The majority indicated that the pandemic still significantly impacts on treatment and outcome of surgical emergency patients (83.1% and 78.5%, respectively). As reasons, the collaborators reported decreased case load in ES (44.7%), but patients presenting with more prolonged and severe diseases, especially concerning perforated appendicitis (62.1%) and diverticulitis (57.5%). Otherwise, approximately 50% of the participants still observe a delay in time-to-intervention in ES compared with the situation before the pandemic. Relevant causes leading to enlarged time-to-intervention in ES during the pandemic are persistent problems with in-hospital logistics, lacks in medical staff as well as operating room and intensive care capacities during the pandemic. This leads not only to the need for triage or transferring of ES patients to other hospitals, reported by 64.0% and 48.8% of the collaborators, respectively, but also to paradigm shifts in treatment modalities to non-operative approaches reported by 67.3% of the participants, especially in uncomplicated appendicitis, cholecystitis and multiple-recurrent diverticulitis. Conclusions: The SARS-CoV-2 pandemic still significantly impacts on care and outcome of patients in ES. Well-known problems with in-hospital logistics are not sufficiently resolved by now; however, medical staff shortages and reduced capacities have been dramatically aggravated over last two pandemic years

Membrane topology of ATP synthase from bovine heart mitochondria and Escherichia coli.

The polypeptides exposed to lipids in the membranous F0 sector of the mitochondrial and Escherichia coli ATP synthases were labelled with radioactive photoreactive lipids. Highly resolving gel electrophoretic conditions were used in order to separate all the eighteen components forming the bovine heart mitochondrial enzyme. The hydrophobic labelling was performed on fully active and inhibitor-sensitive ATP synthases. In the mitochondrial enzyme prepared according to Serrano et al. (1976) [J. Biol. Chem. 251, 2453-2461] seven polypeptides of Mr 30500; 11500; 10500; 10000; 9500; 8500 and 4500 were labelled. The major amount of radioactivity was associated with the 30500-Mr component, which is thought to be the adenine nucleotide carrier. In the preparation of Galante et al., (1979) which almost completely lacks this component [J. Biol. Chem. 254, 12372-12378] nine polypeptides of Mr 25000; 21000; 11500; 10500; 10000; 9500; 9200; 8500 and 4500 were labelled. In the ATPase synthase from E. coli the major amount of labelling was associated with subunit b and only a minor portion with subunit c