Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches

Extracellular vesicles (EVs), through their complex cargo, can reflect the state of their cell of origin and change the functions and phenotypes of other cells. These features indicate strong biomarker and therapeutic potential and have generated broad interest, as evidenced by the steady year-on-year increase in the numbers of scientific publications about EVs. Important advances have been made in EV metrology and in understanding and applying EV biology. However, hurdles remain to realising the potential of EVs in domains ranging from basic biology to clinical applications due to challenges in EV nomenclature, separation from non-vesicular extracellular particles, characterisation and functional studies. To address the challenges and opportunities in this rapidly evolving field, the International Society for Extracellular Vesicles (ISEV) updates its 'Minimal Information for Studies of Extracellular Vesicles', which was first published in 2014 and then in 2018 as MISEV2014 and MISEV2018, respectively. The goal of the current document, MISEV2023, is to provide researchers with an updated snapshot of available approaches and their advantages and limitations for production, separation and characterisation of EVs from multiple sources, including cell culture, body fluids and solid tissues. In addition to presenting the latest state of the art in basic principles of EV research, this document also covers advanced techniques and approaches that are currently expanding the boundaries of the field. MISEV2023 also includes new sections on EV release and uptake and a brief discussion of in vivo approaches to study EVs. Compiling feedback from ISEV expert task forces and more than 1000 researchers, this document conveys the current state of EV research to facilitate robust scientific discoveries and move the field forward even more rapidly

Expression Analysis of ARMC3, a Testis-Specific Gene, in Breast Cancer Patients

Background: Breast cancer is the most prevalent cancer among women. Biomarkers that are expressed in tumors play a pivotal role in diagnosis and treatment. Cancer-testis (CT) antigens are predominantly expressed in the testis and also have inappropriate expression in various tumor types. In the case of expression in tumors, they will be used as immunotherapy targets. Objectives: Expression of ARMC3, a CT antigen, was analyzed to determine its potential as a tumor marker for breast cancer. Materials and Methods: Eighty samples including 40 tumor samples and 40 normal adjacent tissue samples, were gathered from the ICBC biobank. RNA extraction was carried out on all samples. The extracted RNA was treated by DNaseI, after which cDNA was synthesized. Expression of ARMC3 with ACTB (internal control) was studied using Real-Time PCR (polymerase chain reaction). Results: Overall, 43.6% of tumors and 25.6% of normal adjacent tissues expressed ARMC3. ARMC3 was overexpressed in 41% of tumor samples (P = 0.00) and showed decreased expression in 46.2% (P = 0.00). Also, the expression of this gene in 12.8% of tumors was unchanged, which was statistically significant. It should be noted that all samples expressed ACTB gene. Conclusions: Expression of ARMC3 in tumor samples and normal adjacent tissue is very important. The expression of this gene in tumor-adjacent tissue may be associated with the stage of cancer; it may be that these tissues are affected by epigenetic and oncogenic changes of breast cancer. Accordingly, aberrant expression of ARMC3 in tumor samples may be an attractive candidate for use as a tumor marker

Quercetin Prevents Body Weight Loss Due to the Using of Superparamagnetic Iron Oxide Nanoparticles in Rat

Background: Superparamagnetic iron oxide nanoparticles (SPION) have been largely considered for numerous applications in biomedicine such as magnetic resonance imaging, hyperthermia, cell tracking, anticancer treatment, and targeted delivery of drugs or genes. However, they may have side effects such body weight loss. Quercetin (QT), a strong antioxidant and free radical scavenger and a natural flavonoid, has a wide range of biological and therapeutic effects. In this study, the effect of QT on prevention of weight loss due to the using of SPION has been investigated. Materials and Methods: SPION and QT-SPION were administered orally at 50 and 100 mg/kg for 7 days. Then, the body weight was measured at the beginning and the end of the study. Results: Rats fed with 50 and 100 mg/kg SPION showed a significant weight loss, whereas those that fed with 50 mg/kg QT-SPION did not. A weight loss was observed in rats treated with 100 mg/kg of QT-SPION. Conclusions: The results of this study showed that quercetin could prevent weight loss due to the SPION

PIWIL2 and PL2L60 (Cancer/Testis genes) Expression in Breast Cancer

Background: Cancer/testis antigens (CTAs) are members of a group of proteins which are normally expressed in testis germ cells and to a lesser extent in the ovaries. Because of recent reports about their aberrant and specific expression in some tumoral tissues, they may play a role as new candidates for targeted therapy. Therefore, the study of the expression pattern of these biomarkers and its relationship with clinical features of the patients is a subject of great interest. Methods: In this study, expression of PIWIL2 and genes was studied by multiplex RT-PCR in 65 breast tissue samples including 30 invasive ductal carcinomas (IDC), 30 normal adjacent tissue samples and five normal breast tissue samples and 2 normal testicular tissue samples as positive controls. beta actin was considered as internal control. Results: Results of gel electrophoresis analysis demonstrated no significant expression of target genes in any sample except testis. Simultaneously, beta actin was expressed in all the samples. Conclusions: The present study indicates lack of PIWIL2 and PL2L60 expression at mRNA level in breast cancer. Although cell lines can be used in cancer research, they are not representative of tumor tissues. More studies investigating the expression of these genes at protein level will help us decide whether to apply these candidate genes as tumor markers or not

Using superparamagnetic iron oxide nanoparticles to enhance bioavailability of quercetin in the intact rat brain

Abstract Background Quercetin (QT) as a bioactive flavonoid has a potential therapeutic activity for numerous neuronal injuries and neurodegenerative diseases. However, the low absorption rate of QT, especially through the blood-brain barrier, restricts its bioactivity in the body. The current research took the advantage of superparamagnetic iron oxide nanoparticles (SPIONs) to enhance the bioavailability of quercetin. Methods Quercetin conjugated with SPIONs was prepared by means of nanoprecipitation method and was characterized by X-ray diffractometer, scanning electron microscope, and Fourier transformed infrared spectrometer analyses. Wistar male rats were orally fed by gavage with QT and QT-SPION at 50 and 100 mg/kg daily doses for 7 days. Using high-performance liquid chromatography (HPLC) method, biodistribution of QT was evaluated in plasma and brain tissue. Results The outcomes of this research revealed a higher concentration in the plasma and brain of the rats fed with QT-SPION in comparison to free QT. Conclusion The results of this study confirm that SPION as a targeted drug delivery system enhances the bioavailability of quercetin in the brain about ten folds higher than free quercetin and could be used for the treatment of neurodegenerative disorders

Ultrasound features of pregnancy‐associated breast cancer: A retrospective observational analysis

Abstract Pregnancy‐associated breast cancer (PABC) is a poor prognosis in women, and the mortality rate is higher in this subgroup of patients than in non‐PABC. This study aims to assess clinicopathological and ultrasound features of patients with PABC. Of 75 patients with breast cancer, 31 cases were in lactating, or pregnancy phase and 44 patients had no recent history of pregnancy/lactation at the time of cancer detection. The available pathological characteristics and ultrasound findings of the PABC and non‐PABC groups were compared. The analysis of ultrasound findings demonstrated that the percentages of antiparallel orientation (p = 0.04) and heterogeneous internal echo pattern (p = 0.002) were higher in the PABC group. The final Breast Imaging Reporting and Data System (BI‐RADS) assessment in the two groups was significantly different (p = 0.008). In this study, most PABCs were BI‐RADS 4c or 5; compared with age‐matched non‐PABC cases. There were significant differences in ER (p = 0.03), receptor groups (p = 0.007), and tumor grade (p = 0.02) in PABC compared to non‐PABC group. To conclude, radiologists should be careful about ultrasound findings of PABC and recommend core needle biopsy in suspected cases