Alginate and cellulose nanofibrils (CNF) are attractive materials for tissue engineering and regenerative medicine. CNF gels are generally weaker and more brittle than alginate gels, while alginate gels are elastic and have high rupture strength. Alginate properties depend on their guluronan and mannuronan content and their sequence pattern and molecular weight. Likewise, CNF exists in various qualities with properties depending on, e.g., morphology and charge density. In this study combinations of three types of alginate with different composition and two types of CNF with different charge and degree of fibrillation have been studied. Assessments of the composite gels revealed that attractive properties like high rupture strength, high compressibility, high gel rigidity at small deformations (Young’s modulus), and low syneresis was obtained compared to the pure gels. The effects varied with relative amounts of CNF and alginate, alginate type, and CNF quality. The largest effects were obtained by combining oxidized CNF with the alginates. Hence, by combining the two biopolymers in composite gels, it is possible to tune the rupture strength, Young’s modulus, syneresis, as well as stability in physiological saline solution, which are all important properties for the use as scaffolds in tissue engineering.

Bakgrund: Lungcancer och malignt melanom är exempel på två sjukdomar som undersöks med dual-modaliteten positron emission tomography/computed tomography (PET/CT). Vid undersökning med PET/CT erhåller patienten både en stråldos från Flourine-18 (18F) märkt med 2-[18F] fluoro-2-deoxy-D-glucose (18FDG) och från CT-modaliteten. Det finns strålningsrisker med undersökningen som kan uttrycka sig i form av stokastiska skador som exempelvis cancer. Syftet med studien var att jämföra stråldoserna mellan lungcancergruppen (misstänkt eller verifierad) och malignt melanomgruppen genom att beräkna den totala effektiva stråldosen samt redovisa riskerna med PET/CT-undersökningen. Material och metod: Materialet omfattades av parametrar gällande undersökningen och urvalet bestod av 20 patienter från lungcancergruppen respektive malignt melanomgruppen som hämtades från Nuklearmedicin, Länssjukhuset Ryhov, Jönköping. En retrospektiv metod med kvantitativ ansats användes för genomförandet av studien. Resultat: En signifikant skillnad (p <0,001) mellan sjukdomsgrupperna förekom där lungcancergruppen erhöll 11,95 milliSievert (mSv) och malignt melanomgruppen 6,03 mSv och den procentuella riskökningen av letal cancer var 0,06 % respektive 0,03 %. Slutsatser: Lungcancergruppen erhöll en dubbelt så hög effektiv dos som malignt melanomgruppen. Den effektiva dosen är dock så låg att riskökningen av letal cancer är marginell och nyttan med undersökningen överväger riskerna. 

Cancer represents a significant global public health challenge and ranks as the second mostcommon cause of death in the United States. The onset of cancer entails an initial phasewhere cells lose their polarity and disconnect from the normal basement membrane, allowingthem to form distinct three-dimensional (3D) configurations that interact with adjacent cellsand the surrounding microenvironment. Cells grown in 2D monolayers demonstrate differentgene expression patterns and different activation of signaling pathways compared to cellscultivated within the natural structure of tumor tissue of the same cell type. Multicellulartumor spheroids (MCTS) are extensively investigated as a well-studied model of organotypiccancer. These spheroids are formed by tumor cells, either alone or in combination with othercell types, and they can be created with or without the application of supportive scaffolds.The MCTSs are also considered promising models for preclinical assessments of chemosensitivity.However, the creation of these tumor spheroids presents challenges, as not alltumor cell lines can consistently form regular spheroids.Cellulose nanofibrils (CNF) have become essential as a sustainable and environmentallyfriendly material. For example, thin films, with inherent mechanical properties, and flexibility,offer versatility across various applications. Also known for its biocompatibility and non-toxicnature, native CNF is a natural option to use. Its fibrous structure closely mimics the collagenmatrix in human tissue, showing potential as an effective scaffold for 3D cell culture. In thisregard, an innovative Layer-by-Layer (LbL) coating technique using CNF-polyelectrolytebilayers was investigated to generate spheroids. This method constructs bilayers of CNFand polyelectrolytes and can coat various surfaces. In this thesis, the first focus was ondemonstrating the spheroid formation capability using low molecular weight polyelectrolytesin LbL assembly. Secondly, an investigation was conducted involving embedding of LbLgrownspheroids in a decellularized extracellular matrix (ECM) aiming to determine howECM, possessing suitable mechanical characteristics, could influence the cancer stem celltraits in spheroids. Thirdly, the thesis demonstrated the utilization of LbL for capturing andreleasing of circulating tumor cells. Lastly, the shift from using low molecular weightpolyelectrolytes in the LbL assembly to high molecular weight counterparts and analyzingthe differences in spheroid formation abilities to assess the underlying differences inmolecular weights of the polyelectrolytes was explored. All-in-all, employing the CNF-basedLbL surface coating strategy explored in the thesis has proven to be promising for thedevelopment of spheroid models closely resembling in vivo conditions and holds significantpotential for applications in drug development.

Three-dimensional (3D) tumor spheroids are regarded as promising models for utilization as preclinical assessments of chemo-sensitivity. However, the creation of these tumor spheroids presents challenges, given that not all tumor cell lines are able to form consistent and regular spheroids. In this context, we have developed a novel layer-by-layer coating of cellulose nanofibril–polyelectrolyte bilayers for the generation of spheroids. This technique builds bilayers of cellulose nanofibrils and polyelectrolytes and is used here to coat two distinct 96-well plate types: nontreated/non-sterilized and Nunclon Delta. In this work, we optimized the protocol aimed at generating and characterizing spheroids on difficult-to-grow pancreatic tumor cell lines. Here, diverse parameters were explored, encompassing the bilayer count (five and ten) and multiple cell-seeding concentrations (10, 100, 200, 500, and 1000 cells per well), using four pancreatic tumor cell lines—KPCT, PANC-1, MiaPaCa-2, and CFPAC-I. The evaluation includes the quantification (number of spheroids, size, and morphology) and proliferation of the produced spheroids, as well as an assessment of their viability. Notably, our findings reveal a significant influence from both the number of bilayers and the plate type used on the successful formation of spheroids. The novel and simple layer-by-layer-based coating method has the potential to offer the large-scale production of spheroids across a spectrum of tumor cell lines.

Elektrokardiografi (EKG) är en metod för att registrera hjärtats elektriska aktivitet, används för att utreda eventuella patologier i hjärtats retledningssystem. Studien fokuserar på parametrar som är relaterade till förmaksdepolarisationen som på ett EKG speglas av P-vågen. Syftet var att utvärdera och analysera reliabiliteten vid upprepade mätningar av specifika EKG-parametrar samt att undersöka eventuella könsskillnader i en hjärtfrisk population. Studien bestod av 45 forskningspersoner. EKG-undersökningen utfördes vid ett tillfälle, därefter mättes EKG-parametrar manuellt på en EKG-utskrift. För att utvärdera mätningarnas reliabilitet inom och mellan observatörer användes intraklasskorrelationskoefficienten (ICC) och för analys av könsskillnader oparat t-test. ICC-värdet var ≥ 0,9 för P-vågsarea och PWAII. För PDV1 och II samt PTFV1 var ICC mellan 0,75–0,9. För PWPTII var ICC mellan 0,5–0,75, medan det var ≤ 0,5 för PWAV1. ICC varierade mellan 0,5–0,75 för PWPTV1. En jämförelse mellan manuell och automatisk tolkning av EKG-parametrarna visade god överensstämmelse. Det förelåg inga statistiskt signifikanta könsskillnader hos någon av EKG-parametrarna (p>0,05). Majoriteten av EKG-parametrar uppvisade god till utmärkt pålitlighet vid upprepande mätningar både inom och mellan observatörer. Det fanns även god överensstämmelse mellan manuell tolkning och automatisk algoritmtolkning. Inga signifikanta könsskillnader i EKG-parametrar kunde påvisas.

In today’s medical science studies, many tumor markers are being used to monitor cancer cell proliferation, but the number of assays for analysis of these markers are few. The aim of this study was to find an easier and more time-efficient way to measure the activity of a specific tumor marker called tymidine kinase 1 (TK1). This tumor marker is an important enzyme involved in cell proliferation and is a key enzyme in the salvage pathway. TK1 activity is related to the occurrence of hematological malignancies and cell activity and therefore have been used as a marker when monitoring this group of patients in treatment. Measurement of the enzyme activity in this study was performed by using an oligonucleotide assay. Detection of the enzyme activity in whole blood and in plasma has not previously been shown. The TK1 activity measured in whole blood and plasma correlated with TK1 activity measured in serum (R2=0,8651 and R2 =0,9845, respectively). It was found that it is possible to determine the TK1 activity in whole blood but only if the activity was measured on the same day as the blood samples were taken. The results shows that the activity measurement of TK1 in plasma and whole blood can be used as a marker to verify patients' therapy in cancer care. This study is only the beginning and further investigations should be made in the future to determine if the method that is subject to this study has the requested effects.

The H295R in vitro cell line produces the majority of the steroidogenesis, for which reason it is commonly used as a screening tool for endocrine disrupting chemicals. Simultaneous determination of the precursor cholesterol and key steroid hormones could give a broad insight into the mechanistic disruption of the steroidogenesis. Steroid hormones have primarily been extracted from H295R incubation medium by means of liquid-liquid extraction (LLE) and the obtained recoveries and matrix effects have typically not been stated or assessed. In the present study a solid-phase extraction (SPE) method was developed and validated for the simultaneous extraction of cholesterol and five key steroid hormones pregnenolone, 17-hydroxyprogesterone, testosterone, cortisol and aldosterone from H295R incubation medium, and finally detected by LC-MS/MS. Cholesterol was recovered at a level of 55.7%, while steroid hormone recoveries ranged from 98.2 to 109.4%. Matrix effects varied between -0.6% and 62.8%. Intra-day precision was deemed acceptable, but the inter-day precision for pregnenolone and aldosterone exceeded the precision limit of 15% RSD. Although LLE has been the most frequently used extraction method in H295R studies, however, our investigation has shown that SPE may relatively easily extract and recover steroid hormones, potentially replacing LLE.

This tutorial provides an overview on a new technique for sample preparation, microextraction by packed sorbent (MEPS). Not only the automation process by MEPS is the advantage but also the much smaller volumes of the samples, solvents and dead volumes in the system. Other significant advantages such as the speed and the simplicity of the sample preparation process are provided. In this tutorial the main concepts of MEPS will be elucidated. Different practical aspects in MEPS are addressed. The factors affecting MEPS performance will be discussed. The application of MEPS in clinical and pre-clinical studies for quantification of drugs and metabolites in blood, plasma and urine will be provided. A comparison between MEPS and other extraction techniques such as SPE, LLE, SPME and SBSE will be discussed. (C) 2011 Elsevier B.V. All rights reserved.

Microextraction by packed sorbent (MEPS) is a new technique for sample preparation that can be connected on-line with liquid chromatography (LC) or gas chromatography (GC) systems without any modifications. This article describes the use of MEPS in clinical and preclinical studies to quantify different drugs in whole blood samples. MEPS was used to determine cyclophosphamide in mouse blood from preclinical g studies using 20 mu L of blood samples. The interday accuracies and 0 precisions ranged from 107-109% and from 2.0-7.0%, respectively. The determination of four immunosuppressive drugs in human blood by MEPS and liquid chromatography-mass spectrometry (LC-MS) is described. The method showed a good selectivity and sensitivity. The calibration curves for everolimus, sirolimus, and tacrolimus ranged from 0.5 to 50 ng/mL and for cyclosporine from 3.0 to 1500 ng/mL. Intraday precisions for the studied immunosuppressive drugs were 2.0-11.7% and interday precision ranged from 5.1 to 13.7% (CV).

Introduction: Chronic Achilles Tendinopathy (CAT) is a common sports injury that affects a substantial portion of the population. One of the clinical treatments of CAT is ultrasound guided sclerosis (UGS), the efficacy of which is both supported and contested in a numberof previous reports. 

Aim: The aim of the study was to assess current and former CAT diagnosed patients’ experiences of stiffness, pain and function of the Achilles tendon (AT) before and after UGS, in order to determine the treatment efficacy.

Methods: A retrospective study based on a modified Victorian Institute of Sports Assessment – Achilles – Survey (VISA-A-S). VISA-A-S questionnaire, with 123 study participants, allof whom were treated at Uppsala University Hospital between 2016-2020 and who had experienced stiffness, pain, and dysfunction of AT before and after UGS. Statistical analysis was performed Wilcoxon Signed-Rank tests.

Results: The response rate to the survey questionnaire was 62%. In total, 80% of all study participants reported that they were satisfied with the treatment. After UGS, 74% reported shorter period of morning stiffness, 85% had less pain and 84% experienced improved function, as compared to before the treatment (P<0.0001 for all).

Conclusion: The results of this study were both statistically and clinically significant. Based on the results of this retrospective study, it is reasonable to conclude that UGS is an effective clinical procedure in treating CAT. This means that this study confirms conclusions of a part of previous studies on the effectiveness of the UGS treatment, which had indicated that UGS is an effective treatment of CAT.

Biologics are an important part of modern healthcare and are mostly administered parenterally due to the fact that it is the route of administration that avoids degradation of biologics and ensures their systemic exposure. However, there is a need to develop oral drug delivery formulations for local treatment of diseases in the gastrointestinal tract (GI). Colloidal capsules is a formulation that can potentially facilitate oral administration of biologics. There have been studies on colloidal capsules and the various ways of manufacturing them, one of which is “Emulsion-based method”. The aim of this study was to produce colloidal capsules made of silica nanoparticles through emulsion-based method, coat them to study their pH-responsive release and characterize them. Encapsulation of a model protein in the silica colloidal capsules was also attempted. pH-responsive release was not studied due to limited access to the laboratory and, a literature study of articles about colloidal capsules was conducted instead, regarding different aspects of colloidal capsule synthesis and encapsulation of various compunds. Web of science was the database used to find scientific studies that specifically produced colloidal capsules. Colloidal capsules were synthesized using a Pickering-emulsion method. Commercially available SiO₂ nanoparticles were used to form the capsules by ultrasonication.  The hydrodynamic size and capsule morphology were analyzed using dynamic light scattering (DLS) and scanning electron microscopy (SEM), respectively. Zeta potential was measured through electrophoretic light scattering (ELS). Articles for the literature study were found using the “web of science” database. Colloidal capsules were successfully produced, coated and characterized. Additionally, the literature study shows that there diverse colloidal capsule synthesis conditions, model proteins and applications for colloidal capsules.

Due to organ donor shortages, long transplant waitlists, and the complications/limitations associated with auto and allotransplantation, biomaterials and tissue-engineered models are gaining attention as feasible alternatives for replacing and reconstructing damaged organs and tissues. Among various tissue engineering applications, bone tissue engineering has become a promising strategy to replace or repair damaged bone. We aimed to provide an overview of bioactive ceramic scaffolds in bone tissue engineering, focusing on angiogenesis and the effect of different biofunctionalization strategies. Different routes to angiogenesis, including chemical induction through signaling molecules immobilized covalently or non-covalently, in situ secretion of angiogenic growth factors, and the degradation of inorganic scaffolds, are described. Physical induction mechanisms are also discussed, followed by a review of methods for fabricating bioactive ceramic scaffolds via microfabrication methods, such as photolithography and 3D printing. Finally, the strengths and weaknesses of the commonly used methodologies and future directions are discussed.

Histoner är en familj av proteiner som bildar nukleosomer vid packning av DNA i cellkärnan. Komplexet mellan histoner och DNA benämns kromatin. Utöver histoner består kromatinet av proteiner som är icke-histoner vilket bidrar till kromatinstabilitet samt genaktivitet. Histoner, med molekylvikter om 11-21 kDa, omfattar fem huvudklasser: H1, H2A, H2B, H3, H4, som bildar ett komplex med varandra genom jonbindning. Histonkomplexet består av (H2A-H2B) som är flankerade av (H3-H4) tetramerer. Eftersom histoner består av basiska aminosyror såsom arginin och lysin är de positivt laddade, vilket underlättar bindningen med DNA som är negativt laddat. Histoner betraktas som antimikrobiella peptider (AMPs) för att de har förmåga att neutralisera bakteriellt endotoxin. Syftet med studien var att rena upp histoner från vetegroddar och karaktärisera dessa, samt studera deras effekt på bakterietillväxt. Vetegroddar användes eftersom de är rika på kromatin. Metoden för att rena upp histoner var baserad på låg jonstyrka ochutnyttjande av tre olika buffertar samt syra extraktion av kromatin med svavelsyra. För att karaktärisera histoner användes SDS-PAGE elektrofores och för att studera deras bakteriella hämning användes E. coli bakterier. Resultatet visade på ett lågt utbyte av histoner, men dock kunde olika histoners huvudklasser separeras och karaktäriseras genom SDS-PAGE. Vissa prover visade på en bakteriehämmande effekt. Optimering av extraktionen är nödvändig för att öka utbytet och därmed bättre kunna studera histonernas antibakteriella effekt. Förmodligen är det homogeniseringen som begränsar utbytet, och därtill bör man också se över hanteringen av syra extraktet för att förhindra aggregering.

Nyckelord: Antimikrobiella peptider, DNA, histoner, jonstyrka, kromatin, syraextraktion, vetegroddar, upprening

Användning av vaccin har varit en av de mest framgångsrika folkhälsoinsatserna som någonsin genomförts för att förebygga infektionssjukdomar och dödsfall över hela världen. Adjuvans är ett ämne som sätts ofta till vacciner vars syfte är att stimulera immunförsvaret på ett mer effektivt sätt. Adjuvanter är partiklar som fagocyters av makrofager och därmed ökar energibehovet och framkallar förändringar i makrofagernas metabolism. Syftet med detta arbete var att undersöka om aluminium (Al)-adjuvanter påverkar makrofagers metabolism genom att mäta intracellulärt glukos hos makrofager efter inkubering med Al-adjuvant. I denna studie undersöktes makrofagers innehåll av glukos i redan insamlade prover från tre friska donatorer. Humana perifera monocyter differentierade och polariserade till M0-, M1- och M2-makrofager. En luminescens baserad metod användes i denna studie för att undersöka makrofagers innehåll av glukos före och efter inkubation av olika koncentrationer med Al-adjuvanter. Resultaten visade att makrofagers glukosproduktion förändras efter inkubation med Al-adjuvanten. M2-makrofager hade den största ökningen av glukos efter tillsats av låga koncentrationer av Al-adjuvanten. Resultatet av denna studie är preliminärt och ytterligare flera försök behövs för att kunna dra slutsatser om en signifikant ökning av glukoskoncentrationen sker hos makrofager efter exponering och därmed fagocytos av Al-adjuvanter.

Movement disorders forbid many people from enjoying their daily lives. As with other diseases, diagnosis and analysis are key issues in treating such disorders. Computer vision-based motion capture systems are helpful tools for accomplishing this task. However Classical motion tracking systems suffer from several limitations. First they are not cost effective. Second these systems cannot detect minute motions accurately. Finally they are spatially limited to the lab environment where the system is installed. In this project, we propose an innovative solution to solve the above-mentioned issues. Mounting the camera on human body, we build a convenient, low cost motion capture system that can be used by the patient while practicing daily-life activities. We refer to this system as active motion capture, which is not confined to the lab environment. Real-time experiments in our lab revealed the robustness and accuracy of the system.

The spinal cord is an important component in the processing and modulation of painful stimuli. Nerve signals from the periphery are relayed and further conducted to the brain (nociception) in the spinal cord, and the most essential modulation of painful information (antinociception) occurs here. Several neurotransmitters are involved in spinal pain modulation, among them acetylcholine. However, the role of acetylcholine has previously been little investigated.

In the present thesis, the acetylcholine release in the spinal cord was studied in vivo. By using spinal microdialysis on anaesthetised rats, the effects on the intraspinal acetylcholine release of various receptor ligands and analgesic agents were examined. This, together with pain behavioural tests and in vitro pharmacological assays, was used to evaluate the role of acetylcholine in spinal pain modulation. The four studies in this thesis resulted in the following conclusions:

An increased release of spinal acetylcholine is associated with an elevated pain threshold, while a decreased acetylcholine release is associated with hyperalgesia, as seen after systemic treatment with a muscarinic agonist and an antagonist.

Lidocaine is a potent analgesic when given systemically. It was found to produce an increase of intraspinal acetylcholine after intravenous injection of analgesic doses. This effect was attenuated after muscarinic, and abolished after nicotinic, receptor blockade.

Various a₂-adrenergic ligands, associated with nociceptive or antinociceptive effects, were found to affect intraspinal acetylcholine release via action on nicotinic receptors.

Finally, the involvement of spinal acetylcholine in the analgesic effects of aspirin and paracetamol was examined. It was found that spinal acetylcholine could participate in the analgesic effects of aspirin, but not of paracetamol.

The present thesis provides data that clearly demonstrate a relationship between intraspinal acetylcholine and antinociception, and elucidate interactions between acetylcholine and other mechanisms that mediate antinociception in the spinal cord.

The local anesthetic lidocaine suppresses different pain conditions when administered systemically. Part of the antinociceptive effect appears to be mediated via receptor mechanisms. We have previously shown that muscarinic and nicotinic agonists that produce antinociception increase the intraspinal release of acetylcholine. In the present study it was hypothesized that systemically administered lidocaine is acting through the same mechanisms as cholinergic agonists and affects the intraspinal release of acetylcholine. Microdialysis probes were placed in anesthetized rats for sampling of acetylcholine. Ten and 30 mg/kg lidocaine injected intravenously significantly increased the intraspinal release of acetylcholine. The effect of lidocaine could be reduced by pretreatment with intraspinally administered atropine or mecamylamine. Our results suggest that the antinociceptive effect produced by systemically administered lidocaine is mediated through an action on muscarinic and nicotinic receptors.

Abstract:Both systemically and intrathecally administered cholinergic agonists produce antinociception while cholinergic antagonists decrease pain threshold. The mechanism and the site of action of these substances are not known. In the present study it was hypothesized that systemically administered muscarinic agonists and antagonists modify nociceptive threshold by affecting intraspinal release of acetylcholine (ACh). Catheters were inserted into the femoral vein in rats maintained on isoflurane anaesthesia for administration of oxotremorine (10–300 μg/kg) and atropine (0.1, 10, 5000 μg/kg). Spinal microdialysis probes were placed intraspinally at approximately the C2–C5 spinal level for sampling of acetylcholine and dialysis delivery of atropine (0.1, 1, 10 nM). Additionally, the tail-flick behaviour was tested on conscious rats injected intraperitoneally with saline, atropine (10, 100 and 5000 μg/kg), or subcutaneously with oxotremorine (30, 100, 300 μg/kg). Subcutaneous administration of oxotremorine (30, 100, 300 μg/kg) significantly increased the tail-flick latency. These doses of oxotremorine dose-dependently increased the intraspinal release of acetylcholine. Intravenously administered atropine, in a dose that produced hyperalgesia (5000 μg/kg) in the tail-flick test, significantly decreased the intraspinal release of acetylcholine. Our results suggest an association between pain threshold and acetylcholine release in spinal cord. It is also suggested that an approximately 30% increase in basal ACh release produces antinociception and that a 30% decrease in basal release produces hyperalgesia.

Cholinergic agonists produce spinal antinociception via mechanisms involving an increased release of intraspinalacetylcholine. The cholinergic receptor system interacts with several other receptor types, such as a2-adrenergic receptors.To fully understand these interactions, the effects of various receptor ligands on the cholinergic system must be investigatedin detail. This study was initiated to investigate the effects of the a2-adrenergic receptor agonists clonidine and rilmenidineand the a2-adrenergic receptor antagonists yohimbine and efaroxan on spinal cholinergic receptors in the rat. Spinalmicrodialysis was used to measure in vivo changes of acetylcholine after administration of the ligands, with or withoutnicotinic receptor blockade. In addition, in vitro binding properties of the ligands on muscarinic and nicotinic receptorswere investigated. It was found that clonidine and rilmenidine increased, while yohimbine decreased spinal acetylcholinerelease. Efaroxan affected acetylcholine release differently depending on concentration. Nicotinic receptor blockade atten-uated the effect of all ligands. All ligands showed poor binding affinity for muscarinic receptors. On the other hand, allligands possessed affinity for nicotinic receptors. Clonidine and yohimbine binding was best fit to a one site binding curveand rilmenidine and efaroxan to a two site binding curve. The present study demonstrates that the tested a2-adrenergicreceptor ligands affect intraspinal acetylcholine release in the rat evoked by nicotinic receptor mechanisms in vivo, andthat they possess binding affinity to nicotinic receptors in vitro. The binding of a2-adrenergic receptor ligands to nicotinicreceptors might affect the intraspinal release of acetylcholine.

Cadmium telluride (CdTe) semiconductor quantum dots (QDs) are brightly luminescent nanocrystals that have emerged as a new class of fluorescent probes for in vivo bioimaging and theranostic applications. CdTe QDs toxicity to normal human cells is minimized by coating with a less toxic ZnS and ZnSe shell forming a core–shell nanostructure. However, coating with ZnS or ZnSe shell is insufficient to prevent the leaching of toxic Cd metal ions. To further minimize toxicity, thiol dual capped CdTe/CdSe/ZnSe multi-core-multi-shell quantum dots were coated with nanoliposome or liposome vesicles (CdTe/CdSe/ZnSe@liposome) and chitosan nanoparticles (CdTe/CdSe/ZnSe@ChitNPs) and their biocompatibility on HeLa and Vero cells were investigated. Different spectroscopic and microscopic techniques were used to elucidate nanocomposites' optical, morphological, and physicochemical properties. The coating of CdTe/CdSe/ZnSe multi-core-multi-shell quantum dots were conducted at different formulations (F1, F2 and F3) and results from the fluorescence studies show that F3 demonstrated the best interaction for both liposome and ChitNPs composite. Exposure to 12 h UV illumination studies also reveals that CdTe/CdSe/ZnSe@liposome shows an enhancement in fluorescence compared to CdTe/CdSe/ZnSe@ChitNPs. The cytotoxicity of the formulations towards HeLa and Vero cells also depicted minimal toxicity compared to CdTe/CdSe/ZnSe QDs that shows much higher toxicity (IC₅₀ = 0.09381 mg/ml). It was further observed that liposome coated multi-core-multi-shell QDs@F2 demonstrated lower toxicity (IC₅₀ = 0.4364 mg/ml) compared to ChitNPs coated multi-core-multi-shell QDs@F2 (IC₅₀ = 0.1618 mg/ml). Results from the florescence imaging studies reveal that CdTe/CdSe/ZnSe-multi-core-multi-shell QDs liposomes and ChitNPs composite retained most of their fluorescence and properties and could easily be tracked in cells and visualized around the nucleus. This indicates the successful internalization of the QDs in the cytosol. Therefore, these results shows that coating CdTe multi-core-mutli-shell QDs with liposomes and ChitNPs produce better biocompatibility compared to uncoated multi-core–shell QDs. However, liposome coated CdTe/CdSe/ZnSe multi-core-multi-shell quantum dots show better optical properties, photostability and biocompatibility compared to CdTe/CdSe/ZnSe multi-core-multi-shell quantum dots with ChitNPs coating. These particles therefore show good promise in cell-labelling and drug delivery studies.

A series of stiripentol (SIP) analogues namely, 2-1(1E)-1-(1,3-benzodioxol-5-yl)-4,4-dimethylpent-1-en-3-ylidene]-N-(aryl/H)hydrazinecarboxamides 7a-h, (+/-)-(5RS)-N-(aryl/H)-(1,3-benzodioxol-5-yl)-3-tert-butyl-4,5-dihydro-1H-pyrazole-1-carboxamides (+/-)-8a-h, and (+/-)-[(5RS)-(1,3-benzodioxol-5-yl)-3-tert-butyl-4,5-dihydro-1H-pyrazol-1-yl](aryl)methanones (+/-)-13a-f was synthesized by adopting appropriate synthetic routes and was pharmacologically evaluated in the preliminary anticonvulsant screens. The selected bioactive new chemical entities were subjected to ED50 determination and neurotoxicity evaluation. The most active congeners are 7h in MES screen and (+/-)-13b in scPTZ screen which displayed ED50 values of 87 and 110 mg/kg, respectively, as compared to that of STP (ED50 = 277.7 and 115 mg/kg in MES and scPTZ, respectively). (C) 2011 Elsevier Masson SAS. All rights reserved.

<bold>Introduction:</bold> Prostate specific membrane antigen (PSMA), highly expressed in metastatic castration-resistant prostate cancer (mCRPC), is an established therapeutic target. Theranostic PSMA-targeting agents are widely used in patient management and has shown improved outcomes for mCRPC patients. Earlier, we optimized a urea-based probe for radionuclide visualization of PSMA-expression in vivo using computer modeling. With the purpose to develop a targeting agent equally suitable for radionuclide imaging and therapy, the agent containing DOTA chelator was designed (BQ7876). The aim of the study was to test the hypothesis that Lu-177-labeled BQ7876 possesses target binding and biodistribution properties potentially enabling its use for radiotherapy.<bold>Methods:</bold> BQ7876 was synthesized and labeled with Lu-177. Specificity and affinity of [Lu-177]Lu-BQ7876 to PSMA-expressing PC3-pip cells was evaluated and its processing after binding to cells was studied. Animal studies in mice were performed to assess its biodistribution in vivo, target specificity and dosimetry. [Lu-177]Lu-PSMA-617 was simultaneously evaluated for comparison.<bold>Results:</bold> BQ7876 was labeled with Lu-177 with radiochemical yield >99%. Its binding to PSMA was specific in vitro and in vivo when tested in antigen saturation conditions as well as in PSMA-negative PC-3 tumors. The binding of [Lu-177]Lu-BQ7876 to living cells was characterized by rapid association, while the dissociation included a rapid and a slow phase with affinities K-D1 = 3.8 nM and K-D2 = 25 nM. The half-maximal inhibitory concentration for Lu-nat-BQ7876 was 59 nM that is equal to 61 nM for Lu-nat-PSMA-617. Cellular processing of [Lu-177]Lu-BQ7876 was accompanied by slow internalization. [Lu-177]Lu-BQ7876 was cleared from blood and normal tissues rapidly. Initial elevated uptake in kidneys decreased rapidly, and by 3 h post injection, the renal uptake (13 +/- 3%ID/g) did not differ significantly from tumor uptake (9 +/- 3%ID/g). Tumor uptake was stable between 1 and 3 h followed by a slow decline. The highest absorbed dose was in kidneys, followed by organs and tissues in abdomen.<bold>Discussion:</bold> Biodistribution studies in mice demonstrated that targeting properties of [Lu-177]Lu-BQ7876 are not inferior to properties of [Lu-177]Lu-PSMA-617, but do not offer any decisive advantages.

High mammographic density, associated with increased tissue stiffness, is a strong risk factor for breast cancer per se . In postmenopausal women there is no differences in the occurrence of ductal carcinoma in situ (DCIS) depending on breast density. Preliminary data suggest that dense breast tissue is associated with a pro -inflammatory microenvironment including infiltrating monocytes. However, the underlying mechanism(s) remains largely unknown. A major roadblock to understanding this risk factor is the lack of relevant in vitro models. A biologically relevant 3D model with tunable stiffness was developed by cross -linking hyaluronic acid. Breast cancer cells were cultured with and without freshly isolated human monocytes. In a unique clinical setting, extracellular proteins were sampled using microdialysis in situ from women with various breast densities. We show that tissue stiffness resembling high mammographic density increases the attachment of monocytes to the cancer cells, increase the expression of adhesion molecules and epithelia-mesenchymal-transition proteins in estrogen receptor (ER) positive breast cancer. Increased tissue stiffness results in increased secretion of similar pro-tumorigenic proteins as those found in human dense breast tissue including inflammatory cytokines, proteases, and growth factors. ER negative breast cancer cells were mostly unaffected suggesting that diverse cancer cell phenotypes may respond differently to tissue stiffness. We introduce a biological relevant model with tunable stiffness that resembles the densities found in normal breast tissue in women. The model will be key for further mechanistic studies. Additionally, our data revealed several pro-tumorigenic pathways that may be exploited for prevention and therapy against breast cancer.

Background: To investigate whether surface microdialysis (μD) sampling in probes covered by a plastic film, as compared to noncovered and to intraparenchymatous probes, would increase the technique's sensitivity for pathophysiologic events occurring in a liver ischemia-reperfusion model. Placement of μD probes in the parenchyma of an organ, as is conventionally done, may cause adverse effects, e.g., bleeding, possibly influencing outcome.

Methods: A transient ischemia-reperfusion model of the liver was used in six anesthetized normoventilated pigs. μD probes were placed in the parenchyma and on the liver surface. Surface probes were either left uncovered or were covered by plastic film.

Results: Lactate and glucose levels were significantly higher in plastic film covered probes than in uncovered surface probes throughout the ischemic period. Glycerol levels were significantly higher in plastic film covered probes than in uncovered surface probes at 30 and 45 min into ischemia.

Conclusions: Covering the μD probe increases the sensibility of the μD–technique in monitoring an ischemic insult and reperfusion in the liver. These findings confirm that the principle of surface μD works, possibly replacing need of intraparenchymatous placement of μD probes. Surface μD seemingly allows, noninvasively from an organ's surface, via the extracellular compartment, assessment of intracellular metabolic events. The finding that covered surface μD probes allows detection of local metabolic changes earlier than do intraparenchymatous probes, merit further investigation focusing on μD probe design.

Oral administration provides a simple and non-invasive approach for drug delivery. However, due to poor absorption and swift enzymatic degradation in the gastrointestinal tract, a wide range of molecules must be parenterally injected to attain required doses and pharmacokinetics. Here we present an orally dosed liquid auto-injector capable of delivering up to 4-mg doses of a bioavailable drug with the rapid pharmacokinetics of an injection, reaching an absolute bioavailability of up to 80% and a maximum plasma drug concentration within 30 min after dosing. This approach improves dosing efficiencies and pharmacokinetics an order of magnitude over our previously designed injector capsules and up to two orders of magnitude over clinically available and preclinical chemical permeation enhancement technologies. We administered the capsules to swine for delivery of clinically relevant doses of four commonly injected medications, including adalimumab, a GLP-1 analog, recombinant human insulin and epinephrine. These multi-day dosing experiments and oral administration in awake animal models support the translational potential of the system. 

Background: Cystic fibrosis (CF) is a hereditary disorder caused by genetic mutations, which affect the chloride ion channels, leading to disrupted salt balance in different organs. A lack of properly functioning chloride ion channels can lead to formation of thick mucus, which hinders organ function, especially in the lungs where repeated inflammation occurs. Early diagnosis is critical to prevent further deterioration of the patient's condition. Current method of analysis of CF diagnostics uses conductivity meters to measure sweat electrolytes. However, current guidelines suggest using a chloridometer to directly measure chloride concentration, is the most reliable marker of cystic fibrosis. The aim of this project was to conduct a comprehensive evaluation of the new instrument's safety, reliability, validity, and conformity of the reference range to international chloride meter guidelines. Additional aims were to investigate the effect of storage conditions on sweat chloride concentration levels and examine the effect of increased salt intake on sweat test results. The study recruited healthy participants and took samples of their sweat by inducing sweat gland secretion. The chloride ion concentration was determined using a coulometric method.

The results of the study found that the new method was reliable and matched international protocols. It also revealed that an increased salt consumption can impact chloride concentration in sweat, but not to an extent that it can affect medical decisions. Additionally, the study demonstrated that sweat samples can be frozen for up to two weeks without affecting the outcome of the chloride determination. 

During the last decades, Electrical Bioimpedance Spectroscopy (EBIS) has been applied mainly by using the frequency-sweep technique, across a range of many different applications. Traditionally, the tissue under study is considered to be time-invariant and dynamic changes of tissue activity are ignored by treating the changes as a noise source. A new trend in EBIS is simultaneous electrical stimulation with several frequencies, through the application of a multi-sine, rectangular or other waveform. This method can provide measurements fast enough to sample dynamic changes of different tissues, such as cardiac muscle. This high sampling rate comes at a price of reduction in SNR and the increase in complexity of devices. Although the frequency-sweep technique is often inadequate for monitoring the dynamic changes in a variant system, it can be used successfully in applications focused on the time-invariant or slowly-variant part of a system. However, in order to successfully use frequency-sweep EBIS for monitoring time-variant systems, it is paramount to consider the effects of aliasing and especially the folding of higher frequencies, on the desired frequency e.g. DC level. This paper discusses sub-Nyquist sampling of thoracic EBIS measurements and its application in the case of monitoring pulmonary oedema. It is concluded that by considering aliasing, and with proper implementation of smoothing filters, as well as by using random sampling, frequency-sweep EBIS can be used for assessing time-invariant or slowly-variant properties of time-variant biological systems, even in the presence of aliasing. In general, undersampling is not always a problem, but does always require proper consideration.

Apixaban och lågmolekylärt heparin (LMH) är antikoagulantia som förhindrar blodproppsbildning genom att hämma faktor Xa. Allt mer patienter använder apixaban och LMH, vilket gör att laboratoriemedicin på länssjukhuset Ryhov är i behov av att utvärdera analysmetoder för apixaban och LMH för att kunna implementera analyserna i klinisk rutin. Syftet med studien var att utvärdera analysmetoden för bestämning av anti-FXa aktivitet i plasma hos patienter behandlade med apixaban eller LMH med hjälp av kromogen substratmetod. Metodutvärderingen bestod av fyra steg: repeterbarhet, mellanliggande precision, överensstämmelse med validerad metod och analys av normalpopulation. Utvärderingen genomfördes med hjälp av Sysmex CS-2100 där det analyserades 20 respektive 40 patientprover för apixaban och LMH samt 10 normalprover. Aktivitet av faktor Xa bestämdes kvantitativt med användning av ljusabsorption vid 405 nm. Repeterbarhet och mellanliggande precision visade låg CV. Patientprover visade överensstämmande resultat med referensvärden från andra laboratorium där r2 för apixaban och LMH var 0,95. Avvikande resultat kan bero på mätfel eller förväxling mellan prover. Analys av normalpopulation visade att värden låg under det lägsta tillförlitliga värdet. Utvärdering av analysmetoden apixaban och LMH på Ryhovs laboratorium visade goda resultat vilket bekräftar att analysmetoden kan användas i klinisk rutin.

Natural products and their derivatives have and are still used by humans for various health ailments due to their rich sources of drug discovery. New biologically active compounds from natural products play a key role in drug development. Marine sponges and their associated microbes contain a lot of bioactive compounds that are potential for drug development. These compounds produce chemical compounds with useful pharmaceutical properties such as antitumor, anti-infective, anti-inflammatory, and antibacterial properties. The main focus of this project was on the antibacterial activity of six different sponge specimens. The aim was to screen the antibacterial activity of the sponge specimen’s extracts. In order to do so, a Minimum Inhibitory Concentration assay was performed to screen the sponge's antibacterial activity against E. coli and S. aureus. Analytical HPLC was used for separation and Solid Phase Extraction (SPE) was used for determining the effect of salts towards the inhibition of anti-bacterial activity for two selected extracts. Ethanolic extract of Stylissa massa showed antibacterial activity against S. aureus. SPE would be a rapid purification step to remove the salts present in sponges at a high concentration but it has not shown a significant effect on the inhibition of antibacterial activity. However, further separation and purification need to be done to be able to completely screen for all the six different sponge specimens.

Bakgrund: Kallvattenimmersion är en vanlig förekommande metod som är populär inomtränings- och tävlingsvärlden samt terapimiljöer och kan resultera i förbättrat humör. Vidsådana tillfällen kan akuta fysiologiska effekter inträffa, såsom ökad kardiovaskulärarbetsbelastning vilket kan leda till höjt blodtryck och hjärtfrekvens.

Syftet: Syftet med denna studie är att analysera de akuta effekterna avkallvattenimmersion på blodtryck och hjärtfrekvens hos både vana och ovana individersamt att undersöka eventuella skillnader mellan grupperna.

Metod: En kvantitativ studie utfördes på 40 deltagare, varav 20 ovana och 20 vana.Deltagarna inkluderade både män och kvinnor i åldersgruppen 18 till 70 år. Ovanadeltagare exponerades för kallvatten i 30 sekunder medan vana deltagare utsattes i 2minuter.

Resultat: Resultaten visade signifikanta förändringar i både systoliskt och diastolisktblodtryck samt pulstryck hos båda grupperna. Dock observerades ingen signifikantskillnad i hjärtfrekvens hos de vana, utan snarare hos de ovana. Ingen statistisktsignifikant skillnad observerades mellan grupperna vad gäller blodtrycket, däremotnoterades en skillnad i hjärtfrekvensen.

Slutsats: Både vana och ovana deltagare uppvisade ökning i blodtrycket, medan endastde ovana deltagarna uppvisade en ökning i hjärtfrekvens.

Introduktion: Polymerase chain reaction (PCR) är en biokemisk och molekylärbiologisk laboratorieteknik som används för in vitro amplifiering av specifika gensekvenser. Det finns olika varianter av PCR, en mer utvecklad version är Realtids-PCR, även benämndkvantitativ PCR (qPCR). qPCR mäter fluorescensintensitet i varje qPCR cykel. Metoden delas in i fyra huvudfaser: linjär-, tidig exponentiell-, exponentiell- och platåfas.

Syfte: Syftet med projektet var att utvärdera prestanda hos Quantstudio 7 vid varierande reaktionsvolym och plattposition, samt vid singleplex och duplex, för att öka kvaliteten på resultat och göra metoden mer kostnadseffektiv.

Material & metod: Syntetisk DNA-sekvens (gBlock) späddes och sattes upp i en standardkurva med sju punkter och användes för 20 µl respektive 10 µl reaktionsvolymen, varje punkt bestod av 4 replikat. För att utvärdera Duplex vs Singelplex förbereddes standardkurva i kombination med en konstant koncentration av en annan assay. För att undersöka intra-plate variation sattes upp identiska reaktioner i samtliga brunnar i PCR-plattan. 

Resultat: Samtliga experiment gav detekterbara ampliferingsprodukter.  Cq-värdet användes för att beräkna medelvärde och standardavvikelse, samt effektivitet och R2-värde

Slutsats: Resultatet som erhålls från QS instrumentet visade att reaktionsvolymerna 10 µl och 20 µl är jämförbara. Duplex experimentet visade att gener med låg genuttryck kan duplexas med gener som har 10 000x högre genuttryck. Resultatet från intra-plate variation visade att variationen i SD var högre i den högre sidan av PCR-plattan. 

Recent understandings in the development and spread of cancer have led to the realization of novel single cell analysis platforms focused on circulating tumor cells (CTCs). A simple, rapid, and inexpensive analytical platform capable of providing genetic information on these rare cells is highly desirable to support clinicians and researchers alike to either support the selection or adjustment of therapy or provide fundamental insights into cell function and cancer progression mechanisms. We report on the genetic profiling of single cancer cells, exploiting a combination of multiplex ligation-dependent probe amplification (MLPA) and electrochemical detection. Cells were isolated using laser capture and lysed, and the mRNA was extracted and transcribed into DNA. Seven markers were amplified by MLPA, which allows for the simultaneous amplification of multiple targets with a single primer pair, using MLPA probes containing unique barcode sequences. Capture probes complementary to each of these barcode sequences were immobilized on a printed circuit board (PCB) manufactured electrode array and exposed to single-stranded MLPA products and subsequently to a single stranded DNA reporter probe bearing a HRP molecule, followed by substrate addition and fast electrochemical pulse amperometric detection. We present asimple, rapid, flexible, and inexpensive approach for the simultaneous quantification of multiple breast cancer related mRNA markers, with single tumor cell sensitivity.

We analysed and compared MUC7 O-glycosylation and inflammatory biomarkers in saliva from female patients with burning mouth syndrome (BMS) and gender/age-matched controls. Oligosaccharides from salivary MUC7 from BMS and controls were released. Inflammatory mediators were measured by multiplex proximity extension assay. Presence of sialyl-Lewis(x) (Si-Le(x)) epitope on MUC7 was confirmed using Western blot. MUC7 O-glycans and measured inflammatory biomarkers were found to be similar between BMS and controls. However, oligosaccharides sialyl-Lewis(x) (Si-Le(x)) was found to be reduced in samples from BMS patients. Positive correlation (combined patients and controls) was found between levels of C-C motif chemokine 19 (CCL-19) and the amount of core-2 oligosaccharides on MUC7 as well as fractalkine (CX3CL1) and level of sialylation. Patients with BMS were shown to represent a heterogeneous group in terms of inflammatory biomarkers. This indicates that BMS patients could be further stratified on the basis of low-level inflammation. The results furthermore indicate that reduced sialylation of MUC7, particularly Si-Le(x), may be an important feature in patients with BMS. However, the functional aspects and potential involvement in immune regulation of Si-Le(x) remains unclear. Our data suggests a chemokine driven alteration of MUC7 glycosylation.

The continued usage of fossil fuels and non-renewable materials is a key issue in modern economies. To increase the share of sustainable energy sources, bio-based approaches are inevitable. Biofuels have long been a target of the scientific community to replace fossil fuels, however, there is still ongoing research to make them a sustainable alternative. First generation biofuels use so-called energy crops to function as a biomass source for ethanol production. As energy crops would compete for land usage with food crops, first generation biofuels have sparked serious social debate, and are considered to be commercially unviable. However, second generation biofuels solve the above problem by utilising low-value by-products, primarily agricultural waste as a source for biomass.

The beer industry, during the production of fermented beverages, produces massive amounts of by-products, the most abundant being brewer’s spent grain (BSG). During the beer brewing process, after the endosperm of the malted barley is hydrolysed, the resulting solid material is what the industry calls the BSG. This by-product practically consists of the barley bran, which is typically either sold as low-cost animal feed or is disposed of as trash. However, considering the vast amounts of BSG produced by large industrial actors, it poses a great potential as it is mostly lignocellulosic material, and it could be valorised into added-value products, most prominently biofuel by a biorefinery approach.

Historically, glycoside hydrolases have been used to break down recalcitrant lignocellulosic biomass in biorefineries. Lytic polysaccharide monooxygenases (LPMOs) are a novel enzyme class that can induce oxidative cleavage at any distance from the end of the polysaccharide chain. LPMOs represent a great potential in boosting the activity of glycoside hydrolases when used in enzyme cocktails in biorefineries by providing additional chain ends for them to act on. The bacterial LPMO CmAA10 acts on cellulose microfibrils, and its gene was expressed in E. coli for this study. The periplasmic fraction of the microorganism was extracted, purified and identified as CmAA10. With the aid of LPMOs in applying an enzyme cocktail biorefinery approach to BSG, it is hoped to saccharify the cellulosic material and eventually contribute to a future bio-based circular economy.

The actin cytoskeleton plays a fundamental role in eukaryotic cells. Its reorganization is regulated by a plethora of actin-modulating proteins, such as a-actinin. In higher organisms, alpha-actinin is characterized by the presence of three distinct structural domains: an N-terminal actin-binding domain and a C-terminal region with EF-hand motif separated by a central rod domain with four spectrin repeats. Sequence analysis has revealed that the central rod domain of alpha-actinin from the fission yeast Schizosaccharomyces pombe consists of only two spectrin repeats. To obtain a firmer understanding of the structure and function of this unconventional alpha-actinin, we have cloned and characterized each structural domain. Our results show that this alpha-actinin isoform is capable of forming dimers and that the rod domain is required for this. However, its actin-binding and cross-linking activity appears less efficient compared to conventional alpha-actinins. The solved crystal structure of the actin-binding domain indicates that the closed state is stabilised by hydrogen bonds and a salt bridge not present in other a-actinins, which may reduce the affinity for actin.

Objectives : To determine the fit of ceramic inlays manufactured using a recently introduced CAD/CAM-system (Decim) and of two types of laboratory-made heat-pressed ceramics (IPS Empress and Opc).

Materials and methods : Extracted human premolars were prepared to receive mesio-occlusodistal (MOD) ceramic inlays, for which 10 Denzir, 10 IPS Empress, and 10 Opc were fabricated. The Denzir restorations were produced by the manufacturer of the CAD/CAM-system, and the IPS Empress and Opc by student dental technicians. Before luting the internal fit on the diestone models and on the premolars was determined using replicas. After luting on the premolars with a resin composite the marginal and internal fit were measured. The values were analyzed statistically using ANOVA and Scheffe's test at a significance level of p<0.05.

Results : Before luting there were no significant differences ( p>0.05) in the internal gap width between the three systems studied when placed on their matching diestone models. When placed on the premolars a significant difference ( p<0.01) in the internal fit was seen between Empress and Opc before luting, whereas there were no significant differences ( p>0.05) between Empress and Denzir and between Opc and Denzir. Between the diestone models and the premolars there were significant differences ( p<0.01) in the internal fit, except for IPS Empress. After luting there were no significant differences ( p>0.05) between IPS Empress and Denzir, whereas the marginal gap width was significantly wider ( p<0.001) for Opc than for IPS Empress and Denzir. The internal fit was significantly ( p<0.001) wider for Opc than for IPS Empress, whereas there were no significant differences ( p>0.05) between IPS Empress and Denzir or between Opc and Denzir.

Conclusion : After luting there were only slight differences in the fit between the restorations fabricated using the three different manufacturing techniques and ceramics. Therefore, long-term follow-up studies are needed to assess the clinical significance of the slight differences between the three systems.

Boundary cap neural crest stem cells (bNCSCs) are multipotent cells that form a barrier between CNS and PNS, playing an important role in ingrowth of neurites into the spinal cord during development. Because of the stemness and multipotency of bNCSCs, they self-renew and can be used directly for transplantation or as a source of matured neural cells. It is important that cells used for cell therapy differentiate and develop into the mature cells that the recipient needs. To ensure this, cells are guided towards specific cell fates, and one way of doing this is with medium supplements. The purpose of this study was to analyze the effects of three different media with supplements on the differentiation of bNCSCs. Two cell lineages of bNCSCs expressing green- and red fluorescent protein were treated with different media for differentiation. The effects of the media supplements neurotrophic glial cell line-derived neurotrophic factor (GDNF), cilinary neurotrophic factor (CTNF) and fetal bovine serum (FBS) were compared, with one medium containing no additional factors. It was found that when GDNF and CTNF are supplemented in the differentiation media, bNCSCs are guided towards astrocytes. Interestingly, the medium containing no additional factors gave rise to an even amount of neurons and astrocytes. FBS had an inhibitory effect on overall differentiation of bNCSCs, giving rise to the smallest amount of neurons and astrocytes. The bNCSCs are promising for cell therapy, as their differentiation can be guided with the use of medium supplements.