High Throughput Primer Walking of cDNA Clones

Primer walking of cloned DNA is a standard research tool. It has been used in the past to determine the sequence of individual clones of interest. With the expansion of DNA sequencing capacity the need to be able to walk larger numbers of clones has become necessary. Our laboratory is a mid-sized genomics facility. In conjunction with the Advanced Biomedical Computing Center (ABCC) we have developed methods for automating the primer selection, DNA sequencing, contig assembly and sequence analysis for clones arrayed in microtiter format. This approach has allowed us to walk 475 clones (five microtiter plates) selected from a cDNA library.     

MTranslatability

Machine Translation of arbitrary input is difficult,but the output quality can be improved significantlyif writers create documents with MT in mind. Thisarticle deals with ``MTranslatability' – translatabilityof texts by MT systems. It identifies characteristics oftext that decrease MTranslatability and suggests waysto improve them. It also illustrates the effect of writingfor MTranslatability by showing before-and-after picturesof output from various commercially available MT systems,and gives an overview of tools that help identify and correctthe problems.     

Second Life as a support element for learning electronic related subjects: A real case

Looking for more active and motivating methodological alternatives from the students’ perspective, which promote analysis and investigation abilities that make the student a more participative agent and some learning processes are facilitated, a practical study was conducted in the University of La Sabana (Chía, Colombia), in Computing Engineering (INF) and Agroindustrial Production Engineering (IPA) academic programs under the guiding question: Is Second Life an attractive tool for students, does it increment motivation, promote participation and facilitate learning processes in electronic related subjects in Engineering degrees?Second Life (SL) platform was then used as a traditional on-site class complement in electronic related subjects. A follow up was conducted with several groups during a year and a half, some students followed the subject with the traditional on-site methodology and others did it following a mixed methodology that combined traditional on-site lessons with virtual sessions trough SL, making a comparison and applying a quantitative as well as a qualitative methodology for the information analysis and recollection.After the specified study period it was found that using SL as part of the methodology, increments motivation toward electronic related subjects, and promotes participation and investigation, due to the fact that this tool makes learning possible in a funnier, more interactive and deeper way than if it had been taught in a master class. Having real-time access from a virtual classroom to all types of audiovisual information, helps more senses get involved in the learning process, making it more effective; nevertheless a weakness was detected related to the attention and concentration: students tend to get distracted, because their classroom is the computer and from there they have access not only to the many worlds of SL but to all the social networks and the Internet in general.     

Monitoring and improving Greek banking services using Bayesian Networks: An analysis of mystery shopping data

Mystery shopping is a well known marketing technique used by companies and marketing analysts to measure quality of service, and gather information about products and services. In this article, we analyse data from mystery shopping surveys via Bayesian Networks in order to examine and evaluate the quality of service offered by the loan departments of Greek Banks. We use mystery shopping visits to collect information about loan products and services and, by this way, evaluate the customer satisfaction and plan improvement strategies that will assist banks to reach their internal standards. Bayesian Networks not only provide a pictorial representation of the dependence structure between the characteristics of interest but also allow to evaluate, interpret and understand the effects of possible improvement strategies.     

A Novel Human Neutralizing mAb Recognizes Delta,Gamma and Omicron Variants of SARS-CoV-2 and Can Be Used in Combination with Sotrovimab

The dramatic experience with SARS-CoV-2 has alerted the scientific community to be ready to face new epidemics/pandemics caused by new variants. Among the therapies against the pandemic SARS-CoV-2 virus, monoclonal Antibodies (mAbs) targeting the Spike glycoprotein have represented good drugs to interfere in the Spike/ Angiotensin Converting Enzyme-2 (ACE-2) interaction, preventing virus cell entry and subsequent infection, especially in patients with a defective immune system. We obtained, by an innovative phage display selection strategy, specific binders recognizing different epitopes of Spike. The novel human antibodies specifically bind to Spike-Receptor Binding Domain (RBD) in a nanomolar range and interfere in the interaction of Spike with the ACE-2 receptor. We report here that one of these mAbs, named D3, shows neutralizing activity for virus infection in cell cultures by different SARS-CoV-2 variants and retains the ability to recognize the Omicron-derived recombinant RBD differently from the antibodies Casirivimab or Imdevimab. Since anti-Spike mAbs, used individually, might be unable to block the virus cell entry especially in the case of resistant variants, we investigated the possibility to combine D3 with the antibody in clinical use Sotrovimab, and we found that they recognize distinct epitopes and show additive inhibitory effects on the interaction of Omicron-RBD with ACE-2 receptor. Thus, we propose to exploit these mAbs in combinatorial treatments to enhance their potential for both diagnostic and therapeutic applications in the current and future pandemic waves of coronavirus.     

An Overview of the Latest Metabolomics Studies on Atopic Eczema with New Directions for Study

Atopic eczema (AE) is an inflammatory skin disorder affecting approximately 20% of children worldwide and early onset can lead to asthma and allergies. Currently, the mechanisms of the disease are not fully understood. Metabolomics, the analysis of small molecules in the skin produced by the host and microbes, opens a window to observe the mechanisms of the disease which then may lead to new drug targets for AE treatment. Here, we review the latest advances in AE metabolomics, highlighting both the lipid and non-lipid molecules, along with reviewing the metabolites currently known to reside in the skin.     

Hypercholesterolemia in Cancer and in Anorexia Nervosa: A Hypothesis for a Crosstalk

The relationship between cholesterol and cancer has been widely demonstrated. Clinical studies have shown changes in blood cholesterol levels in cancer patients. In parallel, basic research studies have shown that cholesterol is involved in the mechanisms of onset and progression of the disease. On the other hand, anorexic patients have high cholesterol levels and a high susceptibility to cancer. In this review, we first present a brief background on the relations among nutrition, eating disorders and cancer. Using several notable examples, we then illustrate the changes in cholesterol in cancer and in anorexia nervosa, providing evidence for their important relationship. Finally, we show a new possible link between cholesterol disorder in cancer and in anorexia nervosa.     

Chronic Hyperkaliemia in Chronic Kidney Disease: An Old Concern with New Answers

Increasing potassium intake ameliorates blood pressure (BP) and cardiovascular (CV) prognoses in the general population; therefore the World Health Organization recommends a high-potassium diet (90–120 mEq/day). Hyperkalaemia is a rare condition in healthy individuals due to the ability of the kidneys to effectively excrete dietary potassium load in urine, while an increase in serum K⁺ is prevalent in patients with chronic kidney disease (CKD). Hyperkalaemia prevalence increases in more advanced CKD stages, and is associated with a poor prognosis. This scenario generates controversy on the correct nutritional approach to hyperkalaemia in CKD patients, considering the unproven link between potassium intake and serum K⁺ levels. Another concern is that drug-induced hyperkalaemia leads to the down-titration or withdrawal of renin-angiotensin system inhibitors (RASI) and mineralocorticoids receptors antagonists (MRA) in patients with CKD, depriving these patients of central therapeutic interventions aimed at delaying CKD progression and decreasing CV mortality. The new K⁺-binder drugs (Patiromer and Sodium-Zirconium Cyclosilicate) have proven to be adequate and safe therapeutic options to control serum K⁺ in CKD patients, enabling RASI and MRA therapy, and possibly, a more liberal intake of fruit and vegetables.