Simple and Robust Intravital Microscopy Procedures in Hybrid TIE2GFP-BALB/c Transgenic Mice

Purpose

The endeavor of deciphering intricate phenomena within the field of molecular medicine dictates the necessity to investigate tumor/disease microenvironment real-time on cellular level. We, hereby, design simple and robust intravital microscopy strategies, which can be used to elucidate cellular or molecular interactions in a fluorescent mouse model.

Procedures

We crossbred transgenic TIE2GFP mice with nude BALB/c mice, allowing the breeding of immunocompetent and immunodeficient mouse models expressing green fluorescent protein (GFP) in vascular endothelium. Then, we surgically exposed various tissues of interest to perform intravital microscopy.

Results

By utilizing simple tissue preparation procedures and confocal or two-photon microscopy, we produced high-resolution static snapshots, dynamic sequences, and 3D reconstructions of orthotopically grown mammary tumor, skin inflammation, brain, and muscle. The homogenous detection of GFP expressed by endothelial cells and a combination of fluorescence agents enabled landmarking of tumor microenvironment and precise molecular tagging.

Conclusion

Simple intravital microscopy procedures on TIE2GFP mice allowed a real-time multi-color visualization of tissue microenvironment, underlining that robust microscopy strategies are relatively simple and can be readily available for many tissues of interest.

     

Intra-arterial nimodipine for severe cerebral vasospasm after aneurysmal subarachnoid haemorrhage – neurological and radiological outcome

ObjectivesCerebral vasospasm is a known complication to aneurysmal subarachnoid haemorrhage, which can lead to severe morbidity. Intra-arterial vasodilation therapy is widely used as a last resort treatment in patients with symptomatic refractory cerebral vasospasm but there is limited data about the outcome. The purpose of this study is to evaluate the neurological and radiological outcome in patients treated with intra-arterial nimodipine in relation to cerebral infarction, procedure-related complications and clinical outcome.MethodsPatients with refractory cerebral vasospasm treated with intra-arterial nimodipine during 2009–2020 at Sahlgrenska University Hospital were retrospectively reviewed. Neurological outcome (modified Rankin Scale) at 30 days and 6 months, development of cerebral infarction after intra-arterial nimodipine treatment and procedure-related complications were studied.ResultsForty-eight patients were treated with intra-arterial nimodipine. A good outcome (modified Rankin Scale 0–2) was seen in 25% (n = 12) of the patients after 30 days and in 47% (n = 22) of the patients after six months. Infarction related to the vasospastic vessel after treatment with intra-arterial nimodipine was seen in 60% (n = 29) of the patients. A total of 124 procedures with intra-arterial nimodipine were performed where complications were seen in 10 (21%) patients in 10 (8%) procedures. Four (8%) patients died within 30 days.ConclusionsA majority of patients developed an ischaemic cerebral infarction in spite of intra-arterial nimodipine treatment. However, a good clinical recovery was seen in almost half of the patients after 6 months. Minor complications occurred in one out of five patients.     

Effect of Long-Term Hydroxytyrosol Administration on Body Weight,Fat Mass and Urine Metabolomics: A Randomized Double-Blind Prospective Human Study

Hydroxytyrosol (HT) is a natural antioxidant found in olive products and characterized by well-documented beneficial effects on human health. Several research studies are ongoing that aim to investigate its potency and molecular mechanism of action. The present study aimed to investigate the potential effect of HT on human obesity through a randomized double-blind prospective design. HT in two different doses (15 and 5 mg/day) and a placebo capsule was administered to 29 women with overweight/obesity for six months and their weight and fat mass were monitored at three time points (baseline, 4, 12 and 24 weeks). Statistically significant weight and visceral fat mass loss (%weight loss: p = 0.012, %visceral fat loss: p = 0.006) were observed in the group receiving the maximum HT dosage versus placebo after 4 weeks of the intervention, with attenuation of these findings at 12 and 24 weeks of the study. Urine samples were collected during the intervention and analyzed via liquid chromatography–high-resolution mass spectrometry for untargeted metabolomic purposes and comparisons between study groups were performed. HT administration was safe and well-tolerated. To the best of our knowledge, this is the first human cohort investigating the effects of HT on obesity for a prolonged study period.     

Prophylaxis of cancer-associated venous thromboembolism with low-molecular-weight heparin-tinzaparin: Real world evidence

Thromboprophylaxis, as a preventive measure for cancer-associated thrombosis (CAT), may be beneficial for patients with active cancer and high-risk for thrombosis. The present post hoc analysis include a total of 407 patients enrolled in the Greek Management of Thrombosis study, who received thromboprophylaxis with tinzaparin. The objectives of the present analysis were: i) To obtain sufficient evidence for the administration of prophylaxis in patients with active cancer, irrespective of Khorana risk assessment model score; ii) to identify the selection criteria for both dose and duration of tinzaparin; and iii) to evaluate the efficacy and safety of tinzaparin administered for CAT prophylaxis. The main tumor types for the patients included in the present study were as follows: Lung (25.1%), pancreatic (14.3%), breast (9.1%), stomach (8.4%), colorectal (7.9%) and ovarian (7.6%). Furthermore, metastatic disease was observed in 69.5% of the patients. High thrombotic burden agents (HTBAs) were administered to 66.3% of the patients, and 17.4% received erythropoietin. A total of 43.7% of the patients exhibited a Khorana score <2. The results of the present study demonstrated that both the presence of metastatic disease and the use of HTBAs seemed to influence oncologists'' decisions for the use of thromboprophylaxis in patients with active cancer, regardless of Khorana score. Tinzaparin, in dose expressed in the standard notation for heparins, i.e., anti-Xa factor international units (Anti-Xa IU), was administered at an intermediate dose (InterD; 8,000-12,000 Anti-Xa IU; once daily) to 52.4% of patients, while the remaining patients received a prophylactic dose (ProD; ≤4,500 Anti-Xa IU; once daily). The average duration of thromoprophylaxis was 5 months. Furthermore, a total of 14 (3.4%) thrombotic events and 6 (1.5%) minor bleeding events were recorded. A total of four thrombotic events were observed following an InterD treatment of tinzaparin, while 10 thrombotic events were observed following ProD treatment. The present study also demonstrated that an InterD of tinzaparin was administered more frequently to patients with a body mass index >30 kg/m², a history of smoking and a history of metastatic disease, along with administration of erythropoietin. InterD tinzaparin treatment was found to be potentially more efficacious and without safety concerns. The present study is a registered clinical trial (ClinicalTrials.gov code, {"type":"clinical-trial","attrs":{"text":"NCT03292107","term_id":"NCT03292107"}}NCT03292107; registration date, September 25, 2017).     

Transverse myelitis related to COVID-19 infection

Journal of Neurology -     

Laparoscopy in the evaluation of women with unexplained ascites: an invaluable diagnostic tool

STUDY OBJECTIVE: To assess whether laparoscopy is a reliable technique for the investigation of women presenting with ascites and in whom the diagnosis remains obscure. DESIGN: Prospective nonrandomized clinical study (Canadian Task Force classification II-2). SETTING: University Departments of a tertiary referral center. PATIENTS: Women presenting in our institution with ascites in whom the diagnosis remained obscure after an extensive nonoperative diagnostic work-up. INTERVENTION: Undiagnosed cases were submitted to laparoscopy, and selective biopsy specimens were taken for histologic study. MEASUREMENTS AND MAIN RESULTS: Over a 3-year period, 73 patients were admitted to our institution with diffuse ascites. In 9 patients (12.3%), the diagnosis remained obscure, and these patients were further investigated with laparoscopy. Selective biopsy specimens obtained at laparoscopy clarified the specific cause of the ascites in all 9 patients. Peritoneal carcinomatosis was responsible in 5 patients (a metastatic gastrointestinal tumor in 1 patient, a malignant mesothelioma of the peritoneum in 1 patient, and a serous papillary carcinoma of the peritoneum and of the ovary in 2 and 1 patients, respectively). Three patients were found with miliary peritoneal tuberculosis, and the last patient had an unusual peritoneal reaction to methylene blue after laparoscopic adhesiolysis. CONCLUSION: Laparoscopy is a valuable means of assessing the peritoneal cavity in patients with unexplained ascites, where the primary cause remains unclear. The diagnosis can be accurately made with selective biopsy specimens, and appropriate treatment can be instituted without delay.