Endothelial Cell Response to Combined Photon or Proton Irradiation with Doxorubicin.

Surgery, radiotherapy, and chemotherapy are essential treatment modalities to target cancer cells, but they frequently cause damage to the normal tissue, potentially leading to side effects. As proton beam radiotherapy (PBT) can precisely spare normal tissue, this therapeutic option is of increasing importance regarding (neo-)adjuvant and definitive anti-cancer therapies. Akin to photon-based radiotherapy, PBT is often combined with systemic treatment, such as doxorubicin (Dox). This study compares the cellular response of human microvascular endothelial cells (HMEC-1) following irradiation with photons (X) or protons (H) alone and also in combination with different sequences of Dox. The cellular survival, cell cycle, apoptosis, proliferation, viability, morphology, and migration were all investigated. Dox monotreatment had minor effects on all endpoints. Both radiation qualities alone and in combination with longer Dox schedules significantly reduced clonogenic survival and proliferation, increased the apoptotic cell fraction, induced a longer G2/M cell cycle arrest, and altered the cell morphology towards endothelial-to-mesenchymal-transition (EndoMT) processes. Radiation quality effects were seen for metabolic viability, proliferation, and motility of HMEC-1 cells. Additive effects were found for longer Dox schedules. Overall, similar effects were found for H/H-Dox and X/X-Dox. Significant alterations between the radiation qualities indicate different but not worse endothelial cell damage by H/H-Dox.

Similar additive effects of doxorubicin in combination with photon or proton irradiation in soft tissue sarcoma models.

High-precision radiotherapy with proton beams is frequently used in the management of aggressive soft tissue sarcoma (STS) and is often combined with doxorubicin (Dox), the first-line chemotherapy for STS. However, current treatment approaches continue to result in high local recurrence rates often occurring within the treatment field. This strongly indicates the need of optimized treatment protocols taking the vast heterogeneity of STS into account, thereby fostering personalized treatment approaches. Here, we used preclinical STS models to investigate the radiation response following photon (X) or proton (H) irradiation alone and in combination with different treatment schedules of Dox. As preclinical models, fibrosarcoma (HT-1080), undifferentiated pleiomorphic sarcoma (GCT), and embryonal rhabdomyosarcoma (RD) cell lines were used; the latter two are mutated for TP53. The cellular response regarding clonogenic survival, apoptosis, cell-cycle distribution, proliferation, viability, morphology, and motility was investigated. The different STS cell types revealed a dose-dependent radiation response with reduced survival, proliferation, viability, and motility whereas G2/M phase arrest as well as apoptosis were induced. RD cells showed the most radiosensitive phenotype; the linear quadratic model fit could not be applied. In combined treatment schedules, Dox showed the highest efficiency when applied after or before and after radiation; Dox treatment only before radiation was less efficient. GCT cells were the most chemoresistant cell line in this study most probably due to their TP53 mutation status. Interestingly, similar additive effects could be observed for X or H irradiation in combination with Dox treatment. However, the additive effects were determined more frequently for X than for H irradiation. Thus, further investigations are needed to specify alternative drug therapies that display superior efficacy when combined with H therapy.

Delayed Diagnosis of Intestinal Tuberculosis: A Case Report.

Abdominal tuberculosis is an uncommon clinical entity, and it can involve the gastrointestinal tract but also the peritoneum, lymph nodes, and solid organs. Its prevalence is higher among individuals from endemic regions for tuberculosis. Epidemiological risk factors associated with typical symptoms and complementary exams should prompt early treatment. We describe the case of a 47-year-old man, originally from India, residing in Portugal for approximately a year. He presented to our emergency department with a three-week-long history of diarrhea, diffuse abdominal pain, more intense on the left quadrants of the abdomen, anorexia, asthenia, and loss of nearly 10% of his body weight. Abdominal and pelvic imaging showed diffuse circumferential thickening of the distal ileum and adjacent mesentery with associated lymphadenopathies. A colonoscopy confirmed the presence of an ulcerated deformative lesion of the cecum with the involvement of the terminal ileum. Initial suspicion of infectious colitis versus inflammatory bowel disease led the team to prescribe antibiotics and corticosteroid therapy, which was associated with bronchoalveolar lavage and sputum samples negative for Mycobacterium tuberculosis, delaying the diagnosis of intestinal tuberculosis. The lack of improvement after weeks of the initial medical therapy, and with histopathological examination of cervical lymphadenopathy showing the presence of granulomatous lymphadenitis with necrosis, led the medical team to start antituberculostatic therapy. The patient showed significant clinical and laboratory improvement, but after two months of adequate treatment a cavitated nodule appeared on the upper lobe of the left lung, and a Mycobacterium tuberculosis complex was identified in the bronchoalveolar lavage. Timely diagnosis and adequate treatment are essential to lower mortality rates of intestinal tuberculosis, and epidemiological risk factors have a great deal of importance on this matter and must always be taken into account.

Primary Sternal Osteomyelitis Caused by Methicillin-Sensitive Staphylococcus aureus: A Diagnosis Rare in Healthy Adults.

Primary sternal osteomyelitis (PSO) is a rare clinical entity, and usually, it is associated with predisposing factors such as intravenous drug use, diabetes mellitus, or human deficiency virus infection. In an otherwise healthy adult, it becomes an even rarer entity. Early diagnosis and treatment minimize associated morbidity, like the need for surgical debridement, longer courses of medication, and length of in-hospital stay. We describe the case of a 54-year-old man without any predisposing risk factors for PSO, who presented with chest pain, erythema, tenderness, and warmth at the right parasternal region. A non-enhanced thoracic tomography showed a 33 mm suspicious pulmonary nodule and no signs of sternum abnormalities. To better evaluate this finding, a positron emission tomography with fluorine-18 fluorodeoxyglucose was performed, showing abnormal uptake of the radionuclide at the sternomanubrial synchondrosis and no abnormal uptake at the lung parenchyma. The presence of Staphylococcus aureus in blood cultures, in conjunction with these results, supported the diagnosis of PSO. The patient completed six weeks of microbiologically oriented antibacterial therapy with complete recovery.

Monogenic diabetes: a new pathogenic variant of HNF1A gene.

Maturity onset diabetes of the young defines a diabetes mellitus subtype, with no insulin resistance or autoimmune pancreatic ß-cells dysfunction, that occurs by mutation in a single gene. A 13-year-old girl hospitalised due to hyperglycemia plus glycosuria without ketosis, and with normal glycated haemoglobin of 6.8%. She started a sugar-free fast-absorption diet and no insulin therapy was required. Fasting glucose was normal, but 2 hours after lunch she presented hyperglycemia as after 2 hours of an oral glucose tolerance test, with 217 mg/dL. Family history was positive for type 2 diabetes mellitus with an autosomal dominant pattern. She was discharged with fast-absorption sugar-free diet and low-dose of sulfonylurea. A genetic test was performed detecting a mutation in heterozygosity of HNF1A gene, compatible with the diagnosis of maturity onset diabetes of the young 3 (MODY3), not reported in the literature. Early recognition of signs and symptoms increase awareness of MODY. Genetic test allows confirmation and leads to optimised treatment.

Occurrence of Guillain-Barre Syndrome During the Initial Symptomatic Phase of COVID-19 Disease: Coincidence or Consequence?

Viral infections are frequently present before the clinical manifestation of Guillain-Barre syndrome (GBS). Multiple studies on coronaviruses have shown that these viruses have neurotropic characteristics, and their molecular mimicry can induce inflammatory demyelinating neuropathy. Herein, we describe a case of a GBS in an 85-year-old patient infected with SARS-CoV-2, manifested with acute progressive symmetric ascending quadriparesis, urinary dysautonomia, and dysphagia, who responded well to treatment with intravenous human immunoglobulin.

CYP21A2 Gene Pathogenic Variants: A Multicenter Study on Genotype-Phenotype Correlation from a Portuguese Pediatric Cohort.

BACKGROUND: Congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency (21OHD) is an autosomal recessive disorder characterized by 3 overlapping phenotypes: salt-wasting (SW), simple virilizing (SV), and non-classic (NC). We aimed at conducting a nationwide genotype description of the CAH pediatric patients and to establish their genotype-phenotype correlation. METHODS: CAH patients were recruited from Portuguese pediatric endocrinology centers and classified as SW, SV, or NC. Genetic analysis was performed by polymerase chain reaction (sequence specific primer, restriction fragment length polymorphism) or direct Sanger sequencing. Genotypes were categorized into 4 groups (0, A, B, and C), according to their predicted enzymatic activity. In each group, the expected phenotype was compared to the observed phenotype to assess the genotype-phenotype correlation. RESULTS: Our cohort comprises 212 unrelated pediatric CAH patients (29% SW, 11% SV, 60% NC). The most common pathogenic variant was p.(Val282Leu; 41.3% of the 424 alleles analyzed). The p.(Val282Leu) variant, together with c.293-13A/C>G, p.(Ile173Asn), p.(Leu308Thr), p.(Gln319*), and large deletions/conversions were responsible for 86.4% of the mutated alleles. Patients' stratification by disease subtype revealed that the most frequent pathogenic variants were c.293-13A/C>G in SW (31.1%), p.(Ile173Asn) in SV (46.9%), and p.(Val282Leu) in NC (69.5%). The most common genotype was homozygosity for p.(Val282Leu; 33.0%). Moreover, we found 2 novel variants: p.(Ile161Thr) and p.(Trp202Arg), in exons 4 and 5, respectively. The global genotype-phenotype correlation was 92.4%. Group B (associated with the SV form) showed the lowest genotype-phenotype correlation (80%). CONCLUSION: Our cohort has one of the largest NC CAH pediatric populations described. We emphasize the high frequency of the p.(Val282Leu) variant and the very high genotype-phenotype correlation observed.

Prospective study of residual neuromuscular block and postoperative respiratory complications in patients reversed with neostigmine versus sugammadex.

BACKGROUND: Postoperative residual curarization (PORC) is an important complication of neuromuscular blocking drug (NMBD) use. This study evaluates the incidence of PORC in the Postanesthesia Care Unit (PACU), its relation to the type of muscle relaxant and reversal agent used, and its implication in the development of postoperative respiratory complications. METHODS: A prospective observational study of cohorts was made with 558 patients operated under general anesthesia. Data regarding patient history, muscle relaxation (train-of-four [TOF]) monitoring, type of relaxants and reversal agents used, and TOF upon admission to the PACU was collected. Four groups were established: cisatracurium, cisatracurium-neostigmine, rocuronium, and rocuronium-sugammadex. An evaluation was made on the incidence of PORC (TOF ratio [TOFr] <0.9) in PACU, and its relation to the relaxants and reversal agents used and to the development of postoperative respiratory complications - minor and major (pneumonia, atelectasis). RESULTS: From a total of 558 patients, 27.9% presented with PORC. According to the NMBD used and the administration or not of a reversal agent, the incidence of PORC in the four groups was: cisatracurium 34%, cisatracurium-neostigmine 28.6%, rocuronium 34%, and rocuronium-sugammadex 1.15%. The global incidence of minor critical respiratory events (CREs) was 14.5%: cisatracurium 27.5%, cisatracurium-neostigmine 17.4%, rocuronium 10.5%, and rocuronium-sugammadex 2.3%, with statistically significant differences. The global incidence of major CREs was 7.5%, and was significantly higher in patients with TOFr<0.9 upon admission to the PACU. CONCLUSIONS: PORC in the PACU is associated to a greater incidence of major and minor respiratory complications. The use of rocuronium-sugammadex significantly reduces the incidence of PORC in the PACU.

[Cigarette smoking in pregnancy: observational prospective study in three towns of Tuscany Region (Central Italy)]. / Fumo di sigaretta in gravidanza: studio osservazionale prospettico in tre città della Toscana.

OBJECTIVE: to assess the prevalence of smoking in pregnancy and its changes after childbirth, and the characteristics associated with a greater likelihood of smoking during pregnancy in a sample of women attending three university hospitals in Tuscany (Central Italy). DESIGN: observational prospective multicentric study. SETTING AND PARTICIPANTS: 1,036 women in ninth month of pregnancy were enrolled at the teaching hospitals of Careggi (Firenze), Pisa and Siena. Women filled a standardized, self-administered questionnaire at enrolment. A second questionnaire was administered by phone to the smoking, ex-smoking and abstinent-during-pregnancy women one year after the delivery. RESULTS: 60.5% of women was never smoker, 17.4% was ex-smoker, 14% of women stopped smoking during pregnancy, and 8.4% were current smokers. Smoking in pregnancy was significantly associated with being younger than 31 years old (OR 1.75; 95%CI 1.01-1.84) and unmarried (OR 1.75; 95%CI 1.10- 2.78), having a low school degree (OR 2.31; 95%CI 1.58-3.36) and a smoking partner (OR 3.03; 95%CI 2.32-3.96). The absolute risk of smoking during pregnancy was 42%. One year after delivery, 44%of women who stopped smoking in pregnancy relapsed. CONCLUSIONS: a not negligible percentage of women residents in Tuscany Region smokes during pregnancy. Relapses after delivery are high. Even if recently in Italy a smoke free legislation was implemented, the study shows a low attention toward passive smoking during pregnancy. Smoking cessation interventions specifically tailored for pregnant women and relapses prevention need to be implemented in Tuscany by all health care professionals who care for women during pregnancy and after delivery.

New aminoacyl-tRNA synthetase-like protein in insecta with an essential mitochondrial function.

Aminoacyl-tRNA synthetases (ARS) are modular enzymes that aminoacylate transfer RNAs (tRNA) for their use by the ribosome during protein synthesis. ARS are essential and universal components of the genetic code that were almost completely established before the appearance of the last common ancestor of all living species. This long evolutionary history explains the growing number of functions being discovered for ARS, and for ARS homologues, beyond their canonical role in gene translation. Here we present a previously uncharacterized paralogue of seryl-tRNA synthetase named SLIMP (seryl-tRNA synthetase-like insect mitochondrial protein). SLIMP is the result of a duplication of a mitochondrial seryl-tRNA synthetase (SRS) gene that took place in early metazoans and was fixed in Insecta. Here we show that SLIMP is localized in the mitochondria, where it carries out an essential function that is unrelated to the aminoacylation of tRNA. The knockdown of SLIMP by RNA interference (RNAi) causes a decrease in respiration capacity and an increase in mitochondrial mass in the form of aberrant mitochondria.