Urine qRT-PCR assay as a screening tool for the detection of congenital human cytomegalovirus infection of infants

Congenital cytomegalovirus (CMV) infection is the most common infectious cause of birth defects. It may cause both, immediate and long term health problems in infants. These include variety of symptoms, such as hearing loss, microcephaly, jaundice, hepatosplenomegaly and seizures. In more severe cases CMV infection can cause the death of an unborn baby and loss of pregnancy. Despite being one of the most extensively studied vertically transmitted infections recently, the adverse effects of vertically transmitted CMV infection are still not well presented to the general public, resulting in a low awareness among potential expectant mothers in Bosnia and Herzegovina. This study aims to elucidate the sensitivity of urine samples for CMV detection in infants as well as to reflect the importance of quantitative real-time PCR (qRT-PCR) in diagnostics of CMV infection in infants. qRT-PCR was used in this study as a technique for the screening of CMV DNA in a cohort of patients based in Sarajevo Canton. These results have shown that urine samples are sufficiently sensitive for the detection of CMV DNA in infants. Furthermore, the simultaneous analysis of several patients has shown a higher number of CMV DNA copies amplified in urine compared to blood samples, derived from the same patient, thus proposing urine as a reliable sample of choice for congenital CMV diagnostics. These findings may propose a need to classify qRT-PCR CMV test among one of the recommended first-trimester pregnancy screening tests, which could help in early detection of CMV infection in Sarajevo Canton

Managers vs. Students: New Approach in Analyzing Current Practices in Capital Structure Management Education

According to Graham and Harvey (2001), an immense gap exists between capital structure theories and practice. This paper argues that this can be partially explained by current educational practices by analyzing undergraduate students'perceptions of capital structure theories and the differences between their opinion and that of the current CEO's and managers. Educators mostly focus on one or maybe two most popular theories and students have much smaller knowledge about other theories. Secondly educational practices favor trade-off theory to asymmetric information based theories. The paper provides some suggestions regarding capital structure education and future research

THE ANALYSIS OF ANTIBIOTIC CONSUMPTION WITHIN THE TERTIARY HEALTHCARE INSTITUTION IN SERBIA DURING 10-YEAR PERIOD (2001-2010)

Objective: To present conclusions related to the antibiotic drug consumption in one tertiary healthcare institution in Serbia in a 10-year period (2001-2010).Methods: It has been analyzed the issue of antibiotics for prevention and treatment of patients hospitalized at the Military Medical Academy between 2001 and 2010. Antibiotic consumption was expressed as a number of Defined Daily Doses per 100 bed-days (DDD/100BD).Results: Total antibiotic consumption ranged from 49.6 DDD/100BD in 2001 up to 60.4 DDD/100BD in 2005. The leading group of antibiotic was third-generation cephalosporins which accounted for 15.1 DDD/100BD of its maximum consumption in 2007. Ceftriaxone was the most frequently used.Conclusion: Analysis of antibiotic consumption, and multidisciplinary approach, has a crucial importance for further survey of protocols compliance concerning the antibiotic consumption in one hospital.Keywords: Antibiotics, Hospital consumption, ATC, DDD, Bacterial resistance

Characterization of solvents and optimization of stability and solubility of bioactive compounds used in lymphoma cell culture treatments

Phytochemicals represent one of the rising agents in cancer research today. They are recognized as one the most abundant bio-active compounds found in many different sources, specifically in plants. Their anti-cancer effects have been frequently explored and reported in various research studies. However, the bioavailability and solubility of phytochemicals still represent the major issue in in vivo and in vitro research. This report analyses the effects of three different phytochemicals, commonly used for survival interference in malignant cancer clones, including thymoquinone (TQ), curcumin, and quercetin in a model of diffuse large B cell lymphoma (DLBCL). In order to characterize the impact of solubility of these compounds to their bioactivity in the DLBCL model, three different but highly widespread solvents were used. Determination of an optimal compound - solvent association is warranted when assessing the stability and activity of phytochemicals in cells. The results of this study indicate the dose-dependent decrease in cellular viability, including the treatments with all combinations of substances and solvents. In addition, we demonstrated that the choice of the solvent greatly influenced solubility and the overall effect of the compound in cancer cells

Micro- and nano-encapsulation in food industry

Encapsulation can be defined as a process of entrapping one substance within another substance producing particles with diameters of a few nm to a few mm. The entrapped material is usually a liquid, but may be a solid or a gas. The main reason of using encapsulation is the fact that some nutrients do not remain in the food for a significant amount of time or may react with the other food components causing undesirable effects. It is possible to use micro- and nanoencapsulation techniques. The first one, microencapsulation, is a technology that can improve the retention time of the nutrient in the food and allow controlled release at specific times, during food consumption or in the intestinal gut (microencapsulation of vitamin). Nanoencapsulation has the potential to protect sensitive bioactive food ingredients from unfavourable environmental conditions, enhance solubilisation, improve taste and odour masking, and enhance bioavailability of poorly absorbable function ingredients. In this review, some relevant aspects of encapsulation methodologies, coating materials and their uses in food technology were discussed

The Establishment of a Primary Culture System of Proximal Tubule Segments Using Specific Markers from Normal Mouse Kidneys

The proximal tubule contains the highest expression of angiotensinogen mRNA and protein within the kidney and plays a vital role in the renal renin-angiotensin system. To study the regulation of angiotensinogen expression in the kidney in more detail, the proximal tubule needs to be accurately isolated from the rest of the nephron and separated into its three segments. The purpose of this study was to design a novel protocol using specific markers for the separation of proximal tubule cells into the three proximal tubule segments and to determine angiotensinogen expression in each segment. Kidneys were removed from C57BL/6J mice. The proximal tubules were aspirated from region of a Percoll gradient solution of the appropriate density. The proximal tubule was then separated into its three segments using segment-specific membrane proteins, after which each segment was characterized by a different specific marker (sodium-glucose transporter 2 for Segment 1; carbonic anhydrase IV for Segment 2; ecto-adenosine triphosphatase for Segment 3). The isolation of proximal tubules into three segments was successful, and angiotensinogen mRNA in Segment 2 and 3 and angiotensinogen protein in all three segments were confirmed. This protocol will be helpful for future studies of the detailed mechanisms of the intrarenal renin-angiotensin system

Religiositet i undervisning och i samhället - hur speglar de varandra?

Syftet med vårt arbete har varit att undersöka vad som karakteriserar religiositeten i Sverige, vilka prioriteringar som görs i religionsundervisningen samt hur undervisningen i religion överensstämmer med religiositeten i samhället. För att besvara vår frågeställning utförde vi en kvalitativ undersökning där vi intervjuade sex religionskunskapslärare i gymnasiet. För att på ett tydligare sätt belysa intervjusvaren har vi använt oss av litteratur som först och främst beskriver religiositeten i samhället samt litteratur som på ett kritiskt sätt granskar religionsundervisningen. Utifrån litteraturen kan vi utläsa att kristendomen tappar sitt fotfäste i samhället och att allt fler söker sig till nyandliga rörelser. I undervisningen prioriterar lärarna världsreligionerna eftersom det står i kursplanen och då eleverna i viss mån förväntar sig det. Det framgår även att lärarna bortprioriterar nyandliga rörelser i undervisningen, dels pga. tidsbrist men också för att kursplanen inte fokuserar nämnvärt på dessa rörelser. En av våra slutsatser med arbetet är att undervisningen och kursplanen inte överensstämmer med den religiositet som finns i samhället

MJERNA NESIGURNOST HEMIJSKOG ANALITIČKOG REZULTATA - POREĐENJE EMPIRIJSKOG PRISTUPA I PRISTUPA MODELIRANJA

Sa  izdavanjem standarda ISO/IEC 17025:1999 procjena mjerne nesigurnosti je postala obaveza kompetentnih ispitnih laboratorija. Taj zahtjev je zajednicu analitičara stavio pred težak zadatak u odabiru adekvatnog pristupa za realnu procjenu mjerne nesigurnosti. Klasičan pristup koji je promovirao GUM je bio teško primjenjljiv na kompleksne hemijske analitičke procese, te je pozdravljen moderan empirijski pristup utemeljen standardom ISO/TS 21748:2004. U ovom stručnom radu su navedeni rezultati primjene klasičnog pristupa modeliranja i empirijskog pristupa za procjenu mjerne nesigurnosti rezultata jednostavne volumetrijske metode za određivanje sadržaja srebra sa svrhom procjene realnosti dobijenih kvantifikacija. Ispitivani uzorci su srebrene legure za nakit čiji je kvalitet specificiran tehničkom legislativom i sa tog stanovišta je značajno uz rezultat ispitivanja izraziti njegovu mjernu nesigurnost. U prvom dijelu su izvori nesigurnosti definisani i kvantificirani na način preporučen u QUAM-u. Prilikom kvantifikacije korišteni su dostupni podaci o mjeriteljskim karakteristikama korištene mjerne opreme, a podaci o preciznosti su eksperimentalno procijenjeni. Dobijen je krajnji rezultat za ukupnu mjernu nesigurnost od 1,15 mg/g Ag. Empirijski pristup je zasnovan na eksperimentalnoj procjeni unutarlaboratorijske standardne devijacije reproduktivnosti i procjeni doprinosa od biasa koja je utvrđena poređenjem dobijenog rezultata sa rezultatom referentne (standardne) ispitne metode. Ukupna mjerna nesigurnost dobijena ovim pristupom je 1,18 mg/g Ag te je ustanovljeno da nema bitne razlike u kvantifikacijama koja su proizašla od primjene različitih pristupa za procjenu. Ovaj eksperiment ukazuje da kod pravilnog promišljanja i korištenja raspoloživih podataka nema disproporcije između procjene po ova dva različita pristupa te se laboratorija može odlučiti da odabere onaj za koji posjeduje dovoljno podataka, resursa i znanja

Troubleshooting of RNA isolation methods in Papanicolaou HPV cervical smears

Cervical cancer is one of the leading causes of cancer in women, worldwide. Infection with human papillomavirus (HPV) has been accepted as the primary cause for the development of invasive cervical cancer and its precursor lesions. Despite HPV infection has been proposed as an indispensable factor for cervical cancer development, only a subset of neoplastic lesions with HPV infection persist and progress to invasive cancer. This suggests us that other molecular events are also involved in cancer progression. Aim of this study was to extract mRNA from cytobrush-collected healthy and HPV infected cervical epithelial cells and investigate various RNA extraction and purification protocols for assessment of RNA yield and quality. Taking into consideration that cervical cancer screening is based on the cytology based Papanicolaou test (Pap test), main challenge is to investigate whether the samples obtained by regular Pap testing can be used for gene expression analysis. For this purpose, a total of 68 cervical specimens were previously tested for HPV infection. Following HPV testing, samples were submitted to RNA extraction and compared to the products after additional purification step involving DNase I. Products obtained after different RNA extraction and purification methods were visualized using 2% agarose gel electrophoresis. In conclusion, DNase I based RNA purification represents a necessary step for the assurance of a high-quality extracted RNA used for gene expression analysis studies. Reliance on commercial kits for RNA extraction only, without performing additional purification step can lead to errors in drawing final conclusions and/or to false negative gene expression profiling, affecting the overall diagnostic procedure. According to obtained results, the type of sampling used in this study was not suitable for the subsequent gene expression analysis