Immunoglobulin G Locus Events in Soft Tissue Sarcoma Cell Lines

Recently immunoglobulins (Igs) have been found to be expressed by cells other than B lymphocytes, including various human carcinoma cells. Sarcomas are derived from mesenchyme, and the knowledge about the occurrence of Ig production in sarcoma cells is very limited. Here we investigated the phenomenon of immunoglobulin G (IgG) expression and its molecular basis in 3 sarcoma cell lines. The mRNA transcripts of IgG heavy chain and kappa light chain were detected by RT-PCR. In addition, the expression of IgG proteins was confirmed by Western blot and immunofluorescence. Immuno-electron microscopy localized IgG to the cell membrane and rough endoplasmic reticulum. The essential enzymes required for gene rearrangement and class switch recombination, and IgG germ-line transcripts were also identified in these sarcoma cells. Chromatin immunoprecipitation results demonstrated histone H3 acetylation of both the recombination activating gene and Ig heavy chain regulatory elements. Collectively, these results confirmed IgG expression in sarcoma cells, the mechanism of which is very similar to that regulating IgG expression in B lymphocytes

Artificial intelligence : A powerful paradigm for scientific research

Y Artificial intelligence (AI) coupled with promising machine learning (ML) techniques well known from computer science is broadly affecting many aspects of various fields including science and technology, industry, and even our day-to-day life. The ML techniques have been developed to analyze high-throughput data with a view to obtaining useful insights, categorizing, predicting, and making evidence-based decisions in novel ways, which will promote the growth of novel applications and fuel the sustainable booming of AI. This paper undertakes a comprehensive survey on the development and application of AI in different aspects of fundamental sciences, including information science, mathematics, medical science, materials science, geoscience, life science, physics, and chemistry. The challenges that each discipline of science meets, and the potentials of AI techniques to handle these challenges, are discussed in detail. Moreover, we shed light on new research trends entailing the integration of AI into each scientific discipline. The aim of this paper is to provide a broad research guideline on fundamental sciences with potential infusion of AI, to help motivate researchers to deeply understand the state-of-the-art applications of AI-based fundamental sciences, and thereby to help promote the continuous development of these fundamental sciences.Peer reviewe

Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition)

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. For example, a key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process versus those that measure fl ux through the autophagy pathway (i.e., the complete process including the amount and rate of cargo sequestered and degraded). In particular, a block in macroautophagy that results in autophagosome accumulation must be differentiated from stimuli that increase autophagic activity, defi ned as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (inmost higher eukaryotes and some protists such as Dictyostelium ) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the fi eld understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. It is worth emphasizing here that lysosomal digestion is a stage of autophagy and evaluating its competence is a crucial part of the evaluation of autophagic flux, or complete autophagy. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. Along these lines, because of the potential for pleiotropic effects due to blocking autophagy through genetic manipulation it is imperative to delete or knock down more than one autophagy-related gene. In addition, some individual Atg proteins, or groups of proteins, are involved in other cellular pathways so not all Atg proteins can be used as a specific marker for an autophagic process. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field

Optimiranje mikrovalne ekstrakcije antihepatotoksičnog triterpenoida iz korijena biljke Actinidia deliciosa i usporedba s konvencionalnim metodama ekstrakcije

A simple and rapid microwave-assisted extraction (MAE) procedure has been developed and optimized for extracting triterpenoids (TTP) from the Actinidia deliciosa root. Several variables that could potentially affect the extraction efficiency, namely extraction time (min), ethanol fraction (%), liquid:solid ratio (volume per mass) and microwave power (W) have been optimized by means of response surface methodology (RSM). The results suggest that ethanol concentration and microwave power were statistically most significant factors. The optimal conditions were determined and three-dimensional response surfaces were plotted from the mathematical models. The optimum conditions were ethanol fraction 72.67 %, microwave power 362.12 W, liquid:solid ratio (volume per mass) 15:1 and extraction time 30 min. Under the optimum operating conditions, the percentage of extracted TTP was 84.96 %, and MAE showed significantly higher recoveries than those obtained by the conventional extraction methods (ultrasonic and reflux extraction). In addition, a drastic reduction of the extraction time (30 min vs. 8 h) and solvent consumption (15 vs. 30) was achieved, comparable with that provided by the reflux extraction as a reference method.Razvijen je i optimiran jednostavan i brz postupak mikrovalne ekstrakcije (microwave-assisted extraction – MAE) triterpenoida (TTP) iz korijena biljke Actinidia deliciosa. Metodom odzivnih površina (response surface methodology – RSM) optimirano je nekoliko varijabla koje bi mogle utjecati na učinkovitost ekstrakcije, a to su: vrijeme ekstrakcije (min), udjel etanola (%), omjer tekuće i krute faze (V/m) i snaga mikrovalova (W). Rezultati pokazuju da su udjel etanola i snaga mikrovalova statistički najvažniji faktori. Određeni su optimalni uvjeti i matematičkim modelima izračunate trodimenzionalne površine odziva. Optimalni uvjeti bili su udio etanola 72,67 %, snaga mikrovalova 362,12 W, omjer tekuće i krute faze (V/m) 15:1 i vrijeme ekstrakcije 30 min. Pri optimalnim uvjetima ekstrahirano je 84,96 % TTP, a postupkom mikrovalne ekstrakcije dobiveni su bolji prinosi nego konvencionalnim metodama ekstrakcije (ultrazvučne i ekstrakcije pomoću refluksa). Također je znatno smanjeno vrijeme ekstrakcije (s 8 h na 30 min) i potrošnja otapala (s 30 na 15) u usporedbi s ekstrakcijom pomoću refluksa