Cryptosporidium parvum is an important diarrhoea-associated protozoan, which is difficult to propagate in vitro. In 2017, a report described a continuous culture of C. parvum Moredun strain, in the oesophageal squamous cell carcinoma cell line COLO-680N, as an easy-to-use system for C. parvum propagation and continuous production of oocysts. Here, we report that—using the Köllitsch strain of C. parvum—even though COLO-680N cells, indeed, allowed parasite invasion and early asexual parasite replication, C. parvum proliferation decreased after the second day post infection. Considering recurring studies, reporting on successful production of newly generated Cryptosporidium oocysts in the past, and the subsequent replication failure by other research groups, the current data stand as a reminder of the importance of reproducibility of in vitro systems in cryptosporidiosis research. This is of special importance since it will only be possible to develop promising strategies to fight cryptosporidiosis and its ominous consequences for both human and animal health by a continuous and reliable methodological progress.

Colorectal cancer is one of the deadliest cancer forms because of late detection. Biomarkers can be used as an indication of cancer and plays of late a great role in cancer care, there are however currently no specific biomarkers that can be used for early detection of colorectal cancer. The genes AGBL1, DCPS, PHYKPL and RUNX1 have been suggested to be affected in colorectal cancer and the aim of this study was to investigate the expression of these in patients with colorectal cancer to understand more about the mechanisms involved in cancerogenesis. The gene expression in colorectal tumors and normal tissue was analysed by real-time PCR. Immunohistochemistry and western blot were used to analyse the expression of phosphorylated RUNX1 protein. The gene DCPS was downregulated by half and PHYKPL was downregulated about five times. RUNX1 was upregulated three times in the tumors, however, any active RUNX1-protein could not be detected in the tissue. Further studies are needed on these genes to fully understand their potential role in the disease and as prognostic markers.

Abstrakt Bakgrund: Barn som drabbas med tumörer behöver genomgå strålbehandling. För att uppnå optimala behandlingsresultat krävs sederingar eller generella anestesi. Tidigare studier har indikerat att strålningsspecifika effekter påverkade kognitiva funktioner hos barn. Syfte: Karakterisera biomarkörer för skador orsakade av strålning och undersöka de neurotoxiska effekterna som uppstår vid hög exponering för gammastrålning. Metod: Del1: Design av 8 primrar för musgenerna Hes1, Hes5, Timp1, Aldh11l1 . RNA-pool används från fem olika grupper. QPCR-metoden används för att bestämma effektiviteten hos de 8 primrarna. Del2: Experimenten utfördes på NMRI-möss på postnatal dag 10. Genuttryck undersöktes för 5 kontrollgrupper och 5 exponerade grupper för hög dos gammastrålning 2Gy. DNA-extraktion utfördes på hippocampus och prefrontalcortex från mössens hjärna. qPCR-metoden används för att studera uppreglering och nedreglering av gener. Resultat: Effektiviteten för primrarna Hes 5 = 80%, Hes1:2= 93% vilket uppfyllde kriterierna 80–110%. Resultat för genererad nedreglering av BDNF i hippocampus och uppreglering av Hes 5 och Jagged-1 i prefrontal cortex, vilket visade en signifikant skillnad mellan gruppernas medelvärden. Däremot observerade ingen signifikant skillnad i uttrycket av generna Nrf2, Grin2, Jagged-1 i hippocampus. Slutsats: Strålningsdoser som används för att behandla hjärntumörer hos barn kan påverka hur vissa allmänna uttryck och leda till dynamiska förändringar i uttrycket som styr normal hjärnutveckling.

Quantification of soilborne pathogen inoculum loads is important in both agricultural and wildland settings. Quantitative Polymerase Chain Reaction (qPCR) methods using SYBR Green chemistry have been shown to be useful for quantifying fungal inoculum loads in environmental samples. The purpose of this study was to develop a method to quantify fungal pathogen inoculum loads in soil seed banks using a qPCR method with SYBR Green chemistry. The invasive annual grass Bromus tectorum was chosen for this seed bank study. There were three objectives: 1) to design target-specific primers for three fungal pathogens known to be important in Bromus tectorum seed banks, 2) to develop a procedure for measuring inoculum loads in field samples, including optimization of qPCR standard curves and protocols, for these pathogens, and 3) to perform qPCR using this methodology on a representative set of field samples to quantify pathogen DNA in seed bank soil and surface litter. The three pathogens were chosen for quantification based on their hypothesized roles in Bromus tectorum stand failure: the seed pathogen Pyrenophora semeniperda, an undescribed species of Fusarium seed rot pathogen belonging to the F. tricinctum species group (FTSG), and the newly-described causal agent of bleach blonde syndrome (Clarireedia capillus-albis). Primers designed for each pathogen were shown to be target-specific in tests against each other and 12 other fungal species cultured from B. tectorum seed banks. Subsequently developed standard curves for each pathogen had R2 values > 0.98, efficiencies between 90 and 110 percent, and generally optimal dissociation curves. Inoculum loads were expressed for each pathogen as picograms of DNA per microliter of extracted soil or surface litter. Significant differences in measured inoculum loads were found between the targeted pathogens and between soil and litter samples for each pathogen. The data provided reinforces that the SYBR Green qPCR method provides a potentially useful tool for the study of field seed and seedling diseases across a wide spectrum of both wildland and agronomic applications.

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