Nogo-A antibody delivery through the olfactory mucosa mitigates experimental autoimmune encephalomyelitis in the mouse CNS.

Systemic administration of Nogo-A-neutralizing antibody ameliorates experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis. However, the blood-brain barrier (BBB) is a major obstacle limiting the passage of systemically applied antibody to the CNS. To bypass the BBB, in the present study we tested the intranasal route of administration by targeting the olfactory mucosa with the Nogo-A-blocking antibody 11C7 mAb in myelin oligodendrocyte glycoprotein-induced EAE. Antibodies were specifically administered onto the olfactory mucosa using a microcatheter. Antibody distribution was examined in the CNS by ELISA and light-sheet microscopy. The effects of 11C7 mAb on Nogo-A signaling were assessed by Western blotting. EAE-induced deficits were monitored daily. Demyelination was observed on spinal cord histological sections. Gene expression changes were followed by trancriptomic analyses. A sensitive capture ELISA revealed a rapid and widespread distribution of 11C7 mAb in the CNS, including the olfactory bulb, the cerebellum and the lumbar spinal cord, but not in the CSF. Light-sheet microscopy allowed to observe antibody accumulation in the parenchyma, thus demonstrating nose-to-brain transfer of IgG. At the functional level, the widespread penetration of 11C7 mAb in the CNS, including the thoracolumbar spinal cord, resulted in the improvement of motor symptoms and in the preservation of myelin in the spinal cord of EAE mice. This was accompanied by Nogo-A signaling downregulation, as reflected by the decreased level of phosphorylated cofilin observed by Western blotting in the cerebellum. In the brain of EAE score-matched animals, 11C7 modified the expression of genes that can influence neurotransmission and cognitive functions, independently of the demyelination phenotype in the spinal cord. In conclusion, our data show the feasibility of olfactory mucosa-directed administration for the delivery of therapeutic antibodies targeting CNS antigens in EAE mice.

Ultrasonography and Infrared Thermography as a Comparative Diagnostic Tool to Clinical Examination to Determine Udder Health in Sows.

The aim of the study was to examine whether the use of infrared thermography and ultrasonography can complement or replace the clinical examination of the sows' mammary glands for pathological alterations. Sows of different parities with inconspicuous udders on palpation before and after birth (n = 35) and sows at the time of weaning (n = 107) were examined. Thermal images were taken from both sides of the udder, while ultrasound pictures were taken from four sides of the respective mammary glands. Within three weeks before birth, a statistically significant increase in the average surface temperature of the glands of about 1.54 °C and of the thickness of the parenchyma of about 1.39 cm could be observed. After weaning, in 10.3% of the examined sows, roundish hyperechogenic nodules were detected sonographically in the glands´ parenchyma. The average skin temperature above the nodules was 1.24 °C lower compared to the total skin area of the altered complex. However, scratches on the udder skin showed similar temperature changes. In two sows, the nodules remained undetected during the clinical examination. Therefore, sonography seems to be superior compared to clinical and thermographic investigations, although it proved to be very time-consuming.

Bacteriological investigation of milk samples and biopsies from the mammary gland of healthy sows.

OBJECTIVE: Milk samples from clinically healthy sows often contain a similar bacterial content as samples from mastitic sows. To verify whether contamination during sample collection is a possible reason for this or bacterial content in the mammary gland postpartum is a regular finding, the aim of the present study was to compare the suitability of milk samples and mammary gland biopsies for assessing the bacteriological status of healthy sows. MATERIAL AND METHODS: Twenty-five clinically healthy sows of different parities were selected. The mammary skin and teats were cleaned and disinfected before biopsies and milk samples were taken from the second mammary gland on the left udder side one day postpartum. Needle biopsies were performed after local anaesthesia. Samples were investigated bacteriologically for aerobic bacteria and a semi-quantitative classification of bacterial growth was carried out. Additionally, formalin-fixed paraffin-embedded biopsies from 18 of the healthy sows were stained and scored for inflammatory cells. RESULTS: A low bacterial content could be found in 96 % of milk samples and in 92 % of biopsies from the healthy sows. Both Gram-positive (mostly streptococci and staphylococci) and Gram-negative bacteria (Escherichia coli) were detected. Histopathological examination revealed mild inflammatory cell infiltration with mainly plasma cells and lymphocytes, in rare cases neutrophilic granulocytes. CONCLUSION: Biopsies of the mammary gland provided similar results with regard to bacteriological investigation compared to milk sampling although these were collected under aseptic conditions. Therefore, it is assumed that ubiquitous bacteria are regularly present in the colostrum and in the mammary gland tissue of clinically healthy sows after parturition. Histopathological findings were not entirely uneventful. CLINICAL RELEVANCE: Milk samples and biopsies from the thoroughly cleaned and disinfected mammary gland indicate a regular bacterial load of the gland postpartum; biopsies do not provide advantages with regard to hygienic conditions.

Selective CNS Targeting and Distribution with a Refined Region-Specific Intranasal Delivery Technique via the Olfactory Mucosa.

Intranasal drug delivery is a promising approach for the delivery of drugs to the CNS, but too heterogenous, unprecise delivery methods without standardization decrease the quality of many studies in rodents. Thus, the lack of a precise and region-specific application technique for mice is a major drawback. In this study, a previously developed catheter-based refined technique was validated against the conventional pipette-based method and used to specifically reach the olfactory or the respiratory nasal regions. This study successfully demonstrated region-specific administration at the olfactory mucosa resulting in over 20% of the administered fluorescein dose in the olfactory bulbs, and no peripheral bioactivity of insulin detemir and Fc-dependent uptake of two murine IgG1 (11C7 and P3X) along the olfactory pathway to cortex and hippocampus. An scFv of 11C7 showed hardly any uptake to the CNS. Elimination was dependent on the presence of the IgG's antigen. In summary, it was successfully demonstrated that region-specific intranasal administration via the olfactory region resulted in improved brain targeting and reduced peripheral targeting in mice. The data are discussed with regard to their clinical potential.