Improved In vivo Assessment of Pulmonary Fibrosis in Mice using X-Ray Dark-Field Radiography

Idiopathic pulmonary fibrosis (IPF) is a chronic and progressive lung disease with a median life expectancy of 4-5 years after initial diagnosis. Early diagnosis and accurate monitoring of IPF are limited by a lack of sensitive imaging techniques that are able to visualize early fibrotic changes at the epithelial-mesenchymal interface. Here, we report a new x-ray imaging approach that directly visualizes the air-tissue interfaces in mice in vivo. This imaging method is based on the detection of small-angle x-ray scattering that occurs at the air-tissue interfaces in the lung. Small-angle scattering is detected with a Talbot-Lau interferometer, which provides the so-called x-ray dark-field signal. Using this imaging modality, we demonstrate-for the first time-the quantification of early pathogenic changes and their correlation with histological changes, as assessed by stereological morphometry. The presented radiography method is significantly more sensitive in detecting morphological changes compared with conventional x-ray imaging, and exhibits a significantly lower radiation dose than conventional x-ray CT. As a result of the improved imaging sensitivity, this new imaging modality could be used in future to reduce the number of animals required for pulmonary research studies

Mouse nuclear myosin I knock-out shows interchangeability and redundancy of myosin isoforms in the cell nucleus.

Nuclear myosin I (NM1) is a nuclear isoform of the well-known "cytoplasmic" Myosin 1c protein (Myo1c). Located on the 11(th) chromosome in mice, NM1 results from an alternative start of transcription of the Myo1c gene adding an extra 16 amino acids at the N-terminus. Previous studies revealed its roles in RNA Polymerase I and RNA Polymerase II transcription, chromatin remodeling, and chromosomal movements. Its nuclear localization signal is localized in the middle of the molecule and therefore directs both Myosin 1c isoforms to the nucleus. In order to trace specific functions of the NM1 isoform, we generated mice lacking the NM1 start codon without affecting the cytoplasmic Myo1c protein. Mutant mice were analyzed in a comprehensive phenotypic screen in cooperation with the German Mouse Clinic. Strikingly, no obvious phenotype related to previously described functions has been observed. However, we found minor changes in bone mineral density and the number and size of red blood cells in knock-out mice, which are most probably not related to previously described functions of NM1 in the nucleus. In Myo1c/NM1 depleted U2OS cells, the level of Pol I transcription was restored by overexpression of shRNA-resistant mouse Myo1c. Moreover, we found Myo1c interacting with Pol II. The ratio between Myo1c and NM1 proteins were similar in the nucleus and deletion of NM1 did not cause any compensatory overexpression of Myo1c protein. We observed that Myo1c can replace NM1 in its nuclear functions. Amount of both proteins is nearly equal and NM1 knock-out does not cause any compensatory overexpression of Myo1c. We therefore suggest that both isoforms can substitute each other in nuclear processes

A comprehensive and comparative phenotypic analysis of the collaborative founder strains identifies new and known phenotypes.

The collaborative cross (CC) is a large panel of mouse-inbred lines derived from eight founder strains (NOD/ShiLtJ, NZO/HILtJ, A/J, C57BL/6J, 129S1/SvImJ, CAST/EiJ, PWK/PhJ, and WSB/EiJ). Here, we performed a comprehensive and comparative phenotyping screening to identify phenotypic differences and similarities between the eight founder strains. In total, more than 300 parameters including allergy, behavior, cardiovascular, clinical blood chemistry, dysmorphology, bone and cartilage, energy metabolism, eye and vision, immunology, lung function, neurology, nociception, and pathology were analyzed; in most traits from sixteen females and sixteen males. We identified over 270 parameters that were significantly different between strains. This study highlights the value of the founder and CC strains for phenotype-genotype associations of many genetic traits that are highly relevant to human diseases. All data described here are publicly available from the mouse phenome database for analyses and downloads

Rasyonlarda yem katkı maddesi olarak iki Deniz yosununun (Ulva lactuca, Enteremorpha linza) gökkuşağı alabalığının (Oncorhynchus mykiss) büyüme performansı, yem değerlendirme ve vücut kompozisyonu üzerine etkileri

Bu çalışmada, iki deniz yosunu (Ulva lactuca, Enteremorpha linza) içeren rasyonun gökkuşağı alabalığının büyüme performansı, yem değerlendirmesi ve vücut kompozisyonu üzerine etkisinin belirlenmesi amaçlanmıştır. İki deneme rasyonu % 10 U. lactuca ve %10 E. linza unu içerecek düzeyde formülüze edilmiş, kontrol grubuna ise hiçbir deniz yosunu ilave edilmemiştir. Her bir deneme grubu üç tekerrürden oluşmuş, her bir tekerrürde ortalama ağırlığı 32.960.29 g olan 14 balık yer almıştır. Balıklar günde üç kez elle yemlenmiş ve çalışma 60 gün sürmüştür. Deniz yosunu içermeyen Kontol grubu ile U. lactuca ve E. linza unu ihtiva eden gruplar arasında ağırlık artışı, spesifik büyüme oranı, nispi büyüme oranı ve yemden yararlanma yönünden farklılık istatistiki olarak önemli bulunmuştur (P0.05). % 10 E. linza unu içeren grupta, yem değerlendirmenin en düşük olduğu tespit edilmiştir (P0.05). Yaşama oranı tüm deneme gruplarında % 96’dan %98’e değişim göstermiştir. Görünür net protein alımı, protein etkinlik oranı, günlük yem ve toplam yem alımı U. lactuca ve E. linza içerikli rasyonlarda kontrol grubuna nazaran nispeten daha düşük bulunmuştur (P0.05). Deneme sonunda, balıketinde yapılan ham protein, ham yağ ve ham kül oranı deneme başıyla karşılaştırıldığında tüm deneme gruplarında daha yüksek bulunmuştur (P0.05). Bu çalışma sonuçları, gökkuşağı alabalığı rasyonlarında %10 seviyesinde U. lactuca ve E. linza kullanmanın kontrol grubuna göre, balıkların daha düşük bir büyüme ve yem değerlendirmesine neden olduğunu ortaya çıkarmıştır. Bu yüzden, alabalık rasyonlarında bu deniz yosunlarının, optimum kullanımı için daha fazla araştırma yapmaya gerek vardır.In the present study, it was aimed to determine the effects of diet containing two seaweed species, Ulva lactuca and Enteromorpha linza, on the growth performance, feed utilization and body composition of rainbow trout. Two experimental diets were formulated with the usage of 10% U. lactuca meal and 10% E. linza meal in feed and control group had no seaweed ingredients. Each experiment was triplicate and each group had fourteen fish specimens with an average weight of 32.96±0.29 g. Fish were hand fed three times per day for 60 days. Significant differences were determined in weight gain, specific growth rate, relative growth rate and feed utilization between experimental and control groups (P<0.05). Fish fed with the diet containing 10% E. linza meal had the poorest feed utilization. The survival rate ranged from 96% to 98% in all groups during trial period. Apparent net protein retention, protein efficiency rate, daily dry feed intake and total feed intake were significantly lower in fish groups which fed with the diet containing U. lactuca and E. linza than control group (P<0.05). The final levels of crude protein, crude lipid and crude ash were in higher rates in the body composition all the groups compared when compared to the initial level (P<0.05). The results of the experiment revealed that a diet with U. lactuca and E. linza inclusion at 10% levels resulted in a poorer growth and feed utilization for rainbow trout when compared to those of control group. Hence, more defined experiments seem to be necessary in order to determine the optimum dietary inclusion level of these seaweeds in rainbow trout diets

Prevalence and genotyping of bovine Cryptosporidium species in the Mediterranean and Central Anatolia Region of Turkey

The prevalence of Cryptosporidium species in calves and heifers with relation to diarrhea from several herds was investigated in this study. Fecal samples were collected from 135 and 120 pre-weaned calves and 79 and 130 heifers raised in the Central Anatolia (CAR) and Mediterranean Regions (MR) of Turkey, respectively. A total of 86 post-weaned calves in CAR were also included in the study. For diagnostic comparison, all samples were examined by microscopic examination, SSU rRNA nested PCR and TaqMan real-time PCR for the presence of oocyst and Cryptosporidium DNA. In total, 102 (34.0 %) and 93 (37.2 %) of the examined samples from CAR and MR were found positive for Cryptosporidium DNA with both nested PCR and real-time PCR analyses, respectively with an overall prevalence of 35.5 %. The diagnostic sensitivity and specificity of microscopic examination were determined as 68.7 % and 100.0 % compared to molecular tools, respectively. RFLP and sequence analyses of the SSU rRNA from the PCR products revealed that 138 (70.8 %) out of 195 positive isolates were C. parvum further confirming the species-specific real-time PCR results. Among the remaining 57 (29.2 %) positive isolates, 30 (15.4 %) and 27 (13.8 %) were characterized as C. ryanae and C. bovis, respectively. C. parvum was the dominant species in pre-weaned calves especially with diarrhea while C. bovis and C. ryanae were mostly found in post-weaned calves and heifers. The sequence analyses of the gp60 gene of C. parvum isolates revealed two subtypes (IIaA13G2R1, IIaA14G1R1) belonging to zoonotic family IIa, with IIaA13G2R1 being the most common in diarrheic calves