The Effect of Orally Administered Multi-Strain Probiotic Formulation (Lactobacillus, Bifidobacterium) on the Phagocytic Activity and Oxidative Metabolism of Peripheral Blood Granulocytes and Monocytes in Lambs.

Probiotic feed additives have attracted considerable research interest in recent years because the effectiveness of probiotics can differ across microbial strains and the supplemented macroorganisms. The present study was conducted on 16 lambs divided equally into two groups (C-control and E-experimental). The examined lambs were aged 11 days at the beginning of the experiment and 40 days at the end of the experiment. The diet of group E lambs was supplemented with a multi-strain probiotic formulation (Lactobacillus plantarum AMT14, Lactobacillus plantarum AMT4, Lactobacillus rhamnosus AMT15, and Bifidobacterium animalis AMT30), whereas group C lambs did not receive the probiotic additive. At the beginning of the experiment (day 0) and on experimental days 15 and 30, blood was sampled from the jugular vein to determine and compare: phagocytic activity (Phagotest) and oxidative metabolism (Phagoburst) of peripheral blood granulocytes and monocytes by flow cytometry. An analysis of the phagocytic activity of granulocytes and monocytes revealed significantly higher levels of phagocytic activity (expressed as the percentage of phagocytic cells and mean fluorescence intensity) in lambs that were administered the multi-strain probiotic formulation compared with lambs in the control group. The probiotic feed additive also exerted a positive effect on the oxidative metabolism of both granulocytes and monocytes (expressed as the percentage of oxidative metabolism and mean fluorescence intensity) after stimulation with Escherichia coli bacteria and with PMA (4-phorbol-12-ß-myristate-13-acetate). These findings suggest that the tested probiotic formulation may have a positive effect on the immune status of lambs.

Identifying Predictive Biomarkers of Subclinical Mastitis in Dairy Cows through Urinary Metabotyping.

Mastitis is a significant infectious disease in dairy cows, resulting in milk yield loss and culling. Early detection of mastitis-prone cows is crucial for implementing effective preventive measures before disease onset. Current diagnosis of subclinical mastitis (SCM) relies on somatic cell count assessment post-calving, lacking predictive capabilities. This study aimed to identify metabolic changes in pre-SCM cows through targeted metabolomic analysis of urine samples collected 8 wks and 4 wks before calving, using mass spectrometry. A nested case-control design was employed, involving a total of 145 multiparous dairy cows, with disease occurrence monitored pre- and postpartum. Among them, 15 disease-free cows served as healthy controls (CON), while 10 cows exclusively had SCM, excluding those with additional diseases. Urinary metabolite profiling revealed multiple alterations in acylcarnitines, amino acids, and organic acids in pre-SCM cows. Metabotyping identified 27 metabolites that distinguished pre-SCM cows from healthy CON cows at both 8 and 4 wks before parturition. However, only four metabolites per week showed significant alterations (p < 0.005). Notably, a panel of four serum metabolites (asymmetric dimethylarginine, proline, leucine, and homovanillate) at 8 wks prepartum, and another panel (asymmetric dimethylarginine, methylmalonate, citrate, and spermidine) at 4 wks prepartum, demonstrated predictive ability as urinary biomarkers for SCM risk (AUC = 0.88; p = 0.02 and AUC = 0.88; p = 0.03, respectively). In conclusion, our findings indicate that metabolite testing can identify cows at risk of SCM as early as 8 and 4 wks before parturition. Validation of the two identified metabolite panels is warranted to implement these predictive biomarkers, facilitate early intervention strategies, and improve dairy cow management to mitigate the impact of SCM. Further research is needed to confirm the efficacy and applicability of these biomarkers in practical farm settings.

Progesterone-independent endometrial mRNA expression in dairy cows with clinical or subclinical endometritis.

Up to 50 % of dairy cows fail to resolve uterine involution and develop chronic clinical (CE) or subclinical endometritis (SE) 21 days after calving. Clinical endometritis is associated with purulent discharge, while SE is not associated with overt clinical signs. Along with numerous knowledge gaps related to its pathogenesis, SE does not allow for a straightforward and effective therapy. Therefore, it is crucial to unravel differences in the expression of genes among healthy, CE, and SE cows. This might contribute to the discovery of new drug candidates and, in consequence, a potentially effective treatment. In the present study, cows between 21 and 28 days postpartum (PP) were examined using vaginoscopy for the presence of vaginal discharge and endometrial cytology for the determination of the endometrial polymorphonuclear cell (PMN) percentage. Next, an endometrial biopsy sample was taken to investigate the expression of 13 selected candidate genes by qPCR. Uterine health status was assigned to healthy (absence of abnormal vaginal discharge and ≤5 % PMN, n = 13), SE (absence of abnormal vaginal discharge and >5 % PMN, n = 30), and CE (mucopurulent or purulent vaginal discharge and >5 % PMN, n = 9). At the same time, a blood sample was collected to assess serum progesterone concentration and to categorize cows as low (≤1 ng/mL) or high (>1 ng/mL) in progesterone. High expression of IL1B, IL6, IL17A, CXCL8, PTGES, PTGS1, PTGS2, and INHBA genes and low expression of FST was noted in the endometrium of CE compared to healthy cows. Increased endometrial INHBA expression was observed in both SE and CE compared to healthy cows. Interestingly, greater expression of PTGES and PRXL2B genes and lower expression of PTGS2 were characteristic of SE versus CE or healthy. Among cows with no overt clinical symptoms of uterine disease (healthy and SE), the endometrial expression of IL1 B, CXCL8, and PTGES was greater in cows with high versus low serum progesterone. Several genes were differentially expressed among healthy, SE, and CE cows indicating different pathways for the development of different uterine diseases. In conclusion, we found progesterone-independent SE markers, which suggests that low endometrial PTGS2 expression may be indicative of an inadequate immune response and thus contribute to the pathogenesis of SE.

Extended Reality in Diagnostic Imaging-A Literature Review.

The utilization of extended reality (ER) has been increasingly explored in the medical field over the past ten years. A comprehensive analysis of scientific publications was conducted to assess the applications of ER in the field of diagnostic imaging, including ultrasound, interventional radiology, and computed tomography. The study also evaluated the use of ER in patient positioning and medical education. Additionally, we explored the potential of ER as a replacement for anesthesia and sedation during examinations. The use of ER technologies in medical education has received increased attention in recent years. This technology allows for a more interactive and engaging educational experience, particularly in anatomy and patient positioning, although the question may be asked: is the technology and maintenance cost worth the investment? The results of the analyzed studies suggest that implementing augmented reality in clinical practice is a positive phenomenon that expands the diagnostic capabilities of imaging studies, education, and positioning. The results suggest that ER has significant potential to improve diagnostic imaging procedures' accuracy and efficiency and enhance the patient experience through increased visualization and understanding of medical conditions. Despite these promising advancements, further research is needed to fully realize the potential of ER in the medical field and to address the challenges and limitations associated with its integration into clinical practice.

The Effect of the BCO2 Genotype on the Expression of Genes Related to Carotenoid, Retinol, and α-Tocopherol Metabolism in Rabbits Fed a Diet with Aztec Marigold Flower Extract.

This study investigated the effect of the BCO2 genotype and the addition of Aztec marigold flower extract to rabbit diets on the expression of BCO1, BCO2, LRAT, and TTPA genes in the liver. The levels of lutein, zeaxanthin, ß-carotene, retinol, and α-tocopherol in the liver and blood serum of rabbits, as well as plasma biochemical parameters and serum antioxidant enzyme activities were also determined. Sixty male Termond White growing rabbits were divided into three groups based on their genotype at codon 248 of the BCO2 gene (ins/ins, ins/del and del/del); each group was divided into two subgroups: one subgroup received a standard diet, and the other subgroup was fed a diet supplemented with 6 g/kg of marigold flower extract. The obtained results indicate that the BCO2 genotype may affect the expression levels of BCO1 and BCO2 genes in rabbits. Moreover, the addition of marigold extract to the diet of BCO2 del/del rabbits may increase the expression level of the BCO2 gene. Finally, an increase in the amount of lutein in the diet of rabbits with the BCO2 del/del genotype contributes to its increased accumulation in the liver and blood of animals without compromising their health status or liver function.

Personalized Superficial HDR Brachytherapy-Dosimetric Verification of Dose Distribution with Lead Shielding of Critical Organs in the Head and Neck Region.

BACKGROUND: Surface brachytherapy, usually characterized by a high dose gradient, allows the dose to be precisely deposited in the irradiated area while protecting critical organs. When the lesion is located in the nasal or ocular region, the organ of vision must be protected. The aim of this study was to verify the dose distributions near critical organs in the head and neck region during a brachytherapy procedure using lead shielding of the eye. METHODS: Anthropomorphic phantom using 3D-printing technology was prepared. The doses deposited at a point in the lens of the eye and on the surface of the eyelid, directly under the lead shield were calculated and measured using EBT3 radiochromic films. Comparison of doses planned in the treatment planning system using the TG-43 formalism, TG-186 formalism, and measured were also performed. RESULTS: Comparing the planned and calculated doses with TG186 formalism it can be assumed that the use of lead shields is a method for protecting the organ of vision from the adverse effects of ionizing radiation. CONCLUSIONS: The decision to use a lead shield during facial surface brachytherapy procedures should be made on a patient-by-patient basis and based on model-based calculation methods recommended by TG186.

3D printing of individual skin brachytherapy applicator: design, manufacturing, and early clinical results.

Purpose: One of the main challenges in facial region brachytherapy is fixation of vendor-delivered standard applicators. Reproducibility can be maintained; however, there are frequent problems with applicator fitting to the skin surface in pleated regions. Manually prepared individual moulds require technological facilities and highly-trained staff. This article presents 3D-printed applicator preparation for a particular patient skin brachytherapy, using low-cost equipment and free software. We described applicator preparation in a step-by-step workflow. Material and methods: This study demonstrated preparation of a skin brachytherapy applicator for a challenging recurrent tumor located in the nose bridge. During first visit of patient, fiducial markers were placed to enclose treated region. Patient was computed tomography (CT)-scanned, and reconstruction of target volume and surrounding organs at risk (OARs) were performed using treatment planning system (TPS). In TPS on patient's surface, a 1-cm thick bolus was added as a body of applicator. Inside the bolus, source paths were designed, and pre-plan was prepared. Using Beben - DICOM to standard triangle language (STL) software, the body of applicator and source-paths from pre-planning was transformed into an STL file, which was used as a solid definition in 3D printing. Results: The printed applicator fitted very well, and its' placement was quickly consistent regarding placing and securing. CTV was slightly broader in treatment plan (0.34 cm3 vs. 0.31 cm3), and doses given to CTV were lower, except for V150, which was higher for the realized plan (1.15% vs. 1.83%). All reported doses to OARs were lower in the realized plan. Conclusions: A low-cost 3D printer and widely available PLA filaments seem feasible to produce individual contact applicators for skin brachytherapy. Beben - DICOM to STL software and presented workflow appear to be convenient and simple tool.

Thermal Boost to Breast Tumor Bed-New Technique Description, Treatment Application and Example Clinical Results.

(1) Current breast-conserving therapy for breast cancer consists of a combination of many consecutive treatment modalities. The most crucial goal of postoperative treatment is to eradicate potentially relapse-forming residual cancerous cells within the tumor bed. To achieve this, the HDR brachytherapy boost standardly added to external beam radiotherapy was enhanced with an initial thermal boost. This study presents an original thermal boost technique developed in the clinic. (2) A detailed point-by-point description of thermal boost application is presented. Data on proper patient selection, microwave thermal boost planning, and interstitial hyperthermia treatment delivery are supported by relevant figures and schemes. (3) Out of 1134 breast cancer patients who were administered HDR brachytherapy boost in the tumor bed, 262 were also pre-heated interstitially without unexpected complications. The results are supported by two example cases of hyperthermia planning and delivery. (4) Additional breast cancer interstitial thermal boost preceding HDR brachytherapy boost as a part of combined treatment in a unique postoperative setting was feasible, well-tolerated, completed in a reasonable amount of time, and reproducible. A commercially available interstitial hyperthermia system fit and worked well with standard interstitial brachytherapy equipment.

Identification of Serum-Predictive Biomarkers for Subclinical Mastitis in Dairy Cows and New Insights into the Pathobiology of the Disease.

Targeted direct injection/liquid chromatography coupled to tandem mass spectrometry-based metabolomics was employed to identify metabolite alterations that could differentiate subclinical mastitis (SCM) from control (CON) dairy cows at -8, -4, disease diagnosis, +4 and +8 wks relative to parturition. We identified and measured 128 metabolites in the serum. Univariate analysis revealed significant alterations of serum metabolites at all five time points studied. By applying multivariate analyses including principle component analysis and partial least squares-discriminant analysis, some of the metabolites were found to have the strongest power for discriminating the SCM from CON cows. The top five metabolites with the greatest variable importance in projection values were selected as potential biomarkers for SCM. A set of five serum metabolites including lysine, ornithine, isoleucine, LysoPC a C17:0, and leucine at -8 wks and five other metabolites including lysine, leucine, isoleucine, kynurenine, and sphingomyelin (SM) C26:0 at -4 wks prepartum were determined as predictive biomarkers for SCM, which provided highly predictive capabilities with AUC (area under the curve) at 1.00. Five metabolites including lysine, leucine, isoleucine, kynurenine, and SM C26:1 in the serum were identified as diagnostic biomarkers for SCM with the AUC of 1.00. Moreover, we observed that distinct metabolic pathways were affected in SCM cows including lysine degradation, biotin, cysteine, methionine, and glutathione metabolism, valine, leucine, and isoleucine biosynthesis and degradation, and aminoacyl-tRNA biosynthesis prior to and during the occurrence of the disease. Results of this study showed that metabolomics analyses can be used to identify susceptible cows to SCM starting from -8 and -4 wks prepartum and that blood can be used to diagnose cows with SCM.

Combination of mouse prion protein with detoxified lipopolysaccharide triggers colon genes related to inflammatory, antibacterial, and apoptotic responses.

Previously we observed that bacterial lipopolysaccharide (LPS) was able to instantly convert recombinant murine prion protein (moPrP) from an alpha-helical to a beta-sheet enriched state. The objectives of this study were to evaluate the effects of a single in vitro administration of recombinant moPrP alone or combined with detoxified lipopolysaccharide (D-LPS) on innate immunity and antibacterial gene expression in the colon of male FVB/N mice, under an Ussing chamber system. Results showed that moPrP alone affected the expression of genes related to both toll-like receptor (TLR)- and nod-like receptor (NLR)-signaling as well as pro- and anti-inflammatory responses. moPrP induced a strong antibacterial response with Slpi mRNA over expression (> 9-fold). Combination of moPrP with D-LPS on the mucosal side of the colon induced genes associated with TLR-signaling, apoptosis, and a very strong antibacterial response (> 35-fold Slpi expression). Administration of moPrP on the mucosal side and D-LPS on the serosal side triggered expression of 12 genes related to TLR signaling, apoptosis, and antibacterial responses, as illustrated by overexpression of Slpi by >30-fold. The over expression of Slpi mRNA was further reaffirmed by ELISA and when moPrP was added to the mucosal side and D-LPS on the serosal side, an increased Slpi protein was observed. Application of combined moPrP and D-LPS on the mucosal side significantly increased the Slpi protein. Results of this study demonstrated that moPrP alone or combined with D-LPS affected the expression of various genes related to inflammation, antibacterial, and apoptotic responses.

3D-printed surface applicators for brachytherapy: a phantom study.

PURPOSE: Brachytherapy is a great alternative for restrictive surgical procedures in facial cancers. Moreover, dose distribution is more beneficial compared with teleradiotherapy during treatment of lesions located on anatomical curves. However, repetitiveness of application is the main issue associated with using commercial applicators. The risk of its displacement is very unfavorable due to large dose gradients in brachytherapy. The aim of this study was to develop a process of preparation of applicators using 3D printing technology. MATERIAL AND METHODS: In planning system, circular volumes near the nose, eye, and ear were determined on transverse layers of an anthropomorphic phantom. Next, boluses with a thickness of 5 mm and 10 mm were designed for each of the layers. Channels in the 10 mm bolus were designed in such a way to place the catheters into the layers. Prepared applicators were printed using polylactic acid (PLA) filament. Plans to irradiate the films for their calibration and plans for treatment prepared in the treatment planning system were conducted. A special phantom was created to calibrate the radiochromic films. Dose distribution around the designed applicators was measured in an anthropomorphic phantom using films within the layers of phantom. Comparison of doses was performed with two-dimensional gamma analysis using OmniPro I'mRT software. RESULTS: The obtained results confirmed compliance of the planned and measured doses in 92%; the analysis of gamma parameter showed 1%/1 mm for acceptability level of 95%. Moreover, the initial dosimetric analysis for gamma criteria with 2%/2 mm showed compliance at 99%. CONCLUSIONS: The results of the present study confirm potential clinical usefulness of the applicators obtained with the use of 3D printing for brachytherapy.

A Targeted Serum Metabolomics GC-MS Approach Identifies Predictive Blood Biomarkers for Retained Placenta in Holstein Dairy Cows.

The retained placenta is a common pathology of dairy cows. It is associated with a significant drop in the dry matter intake, milk yield, and increased susceptibility of dairy cows to metritis, mastitis, and displaced abomasum. The objective of this study was to identify metabolic alterations that precede and are associated with the disease occurrence. Blood samples were collected from 100 dairy cows at -8 and -4 weeks prior to parturition and on the day of retained placenta, and only 16 healthy cows and 6 cows affected by retained placenta were selected to measure serum polar metabolites by a targeted gas chromatography-mass spectroscopy (GC-MS) metabolomics approach. A total of 27 metabolites were identified and quantified in the serum. There were 10, 18, and 17 metabolites identified as being significantly altered during the three time periods studied. However, only nine metabolites were identified as being shared among the three time periods including five amino acids (Asp, Glu, Ser, Thr, and Tyr), one sugar (myo-inositol), phosphoric acid, and urea. The identified metabolites can be used as predictive biomarkers for the risk of retained placenta in dairy cows and might help explain the metabolic processes that occur prior to the incidence of the disease and throw light on the pathomechanisms of the disease.

Dosimetric assessment of the impact of low-cost materials used in stereolithography in high-dose-rate brachytherapy.

PURPOSE: 3D printing has become a popular and widely available technique of rapid prototyping. The impact of used materials on the dose distribution has been studied for high energy sources. However, brachytherapy sources emit lower energy photons, and materials used in 3D printing may differ. This study was conducted to analyze the influence of common materials (polylactic acid - PLA and acrylonitrile butadiene styrene - ABS) used in stereolithography. MATERIAL AND METHODS: A 3D-printed phantom was designed, printed, and used to calibrate Gafchromic films. In the next step, a range of 1 mm thick plates of PLA and ABS (from zero to thirty) were inserted between source and detector to measure the impact of studied materials on delivered dose. Measurements were performed using a calibrated radiochromic film and Farmer ionization chamber in water. RESULTS: No statistically significant correlation (p = 0.4854) between the thickness of inserted PLA and the dose delivered to the film was obtained. With ionization chamber, Spearman's rank order test showed a significant correlation (p = 0.00004); however, the correlation was found weak. In case of ABS measurement, a statistically significant (p = 0.0159), yet weak negative correlation was found between the thickness of used material and the dose delivered to the film. On the other hand, a weak statistically significant (p = 0.0212) but positive correlation was found when the dose was measured with Farmer ionization chamber. We find these correlations false, as all measured doses were within the measurement uncertainty range (film ±8.0%, Farmer ±8.8%) from 100% of the prescribed dose. CONCLUSIONS: According to obtained results, with the accuracy of measurement under clinical conditions, the impact of highly filled PLA and ABS printed objects on the dose distribution from an 192Ir source in water can be omitted.

The influence of dietary supplementation with the leucine metabolite ß-hydroxy-ß-methylbutyrate (HMB) on the chemotaxis, phagocytosis and respiratory burst of peripheral blood granulocytes and monocytes in calves.

BACKGROUND: A healthy immune system plays a particularly important role in newborns, including in calves that are far more susceptible to infections (viral, bacterial and other) than adult individuals. Therefore, the present study aimed to evaluate the influence of HMB on the chemotactic activity (MIGRATEST® kit), phagocytic activity (PHAGOTEST® kit) and oxidative burst (BURSTTEST® kit) of monocytes and granulocytes in the peripheral blood of calves by flow cytometry. RESULTS: An analysis of granulocyte and monocyte chemotactic activity and phagocytic activity revealed significantly higher levels of phagocytic activity in calves administered HMB than in the control group, expressed in terms of the percentage of phagocytising cells and mean fluorescence intensity (MFI). HMB also had a positive effect on the oxidative metabolism of monocytes and granulocytes stimulated with PMA (4-phorbol-12-ß-myristate-13-acetate) and Escherichia coli bacteria, expressed as MFI values and the percentage of oxidative metabolism. CONCLUSION: HMB stimulates non-specific cell-mediated immunity, which is a very important consideration in newborn calves that are exposed to adverse environmental factors in the first weeks of their life. The supplementation of animal diets with HMB for both preventive and therapeutic purposes can also reduce the use of antibiotics in animal production.

Serum metabolomics identifies metabolite panels that differentiate lame dairy cows from healthy ones.

INTRODUCTION: Although much is known about lameness application of metabolomics technologies to better understanding its etiology and pathogenesis is of utmost interest. OBJECTIVES: The objective of this study was to investigate serum metabolite alterations in pre-lame, lame and post-lame dairy cows in order to identify potential screening serum metabolite biomarkers for lameness and better understand its pathobiology. METHODS: A combination of direct injection and tandem mass spectrometry (DI-MS/MS) with a reverse-phase liquid chromatography and tandem mass spectrometry (LC-MS/MS) analysis was performed in the serum of six cases of lameness and 20 healthy control cows (CON) at - 8 and - 4 weeks prepartum, at lameness diagnosis week, and at + 4 and + 8 weeks postpartum. RESULTS: Data indicated that pre-lame, lame, and post-lame cows experienced altered concentrations of multiple metabolites. It is interesting to note that throughout the 16-weeks of the study, 7 serum metabolites [e.g., diacyl-phosphatidylcholine (PC aa) C30:0, phosphatidylcholine acyl-alkyl (PC ae) C40:2, sphingomyelin (SM) (OH) C14:1, SM C18:0, isoleucine (Ile), leucine (Leu), and lysine (Lys)] differentiated CON cows from the lame ones. Furthermore, 4 metabolic pathways (i.e., Lys degradation, biotin metabolism, tryptophan (Trp) metabolism, and valine [(Val)-Leu-Ile degradation) were altered in cows with lameness during the onset and progression of the disease. CONCLUSION: Multiple metabolite and pathway alterations were identified in the serum of pre-lame, lame, and post-lame cows that through light into the pathobiology of the disease and that can be used as potential biomarker sets that can predict the risk of lameness in dairy cows.

Milk Metabotyping Identifies Metabolite Alterations in the Whole Raw Milk of Dairy Cows with Lameness.

The objective of this study was to evaluate whether whole raw milk originating from Holstein dairy cows affected by lameness alters its composition. A total of 20 healthy control cows and 6 cows diagnosed with lameness were selected out of 100 sampled cows in a nested case control study at 2 weeks postpartum, and whole raw milk samples were collected and analyzed with direct inject/liquid chromatography-tandem mass spectrometry and nuclear magnetic resonance. In total, 168 metabolites were identified and quantified using an in-house mass spectrometry library. A total of 35 of the identified metabolites decreased versus control cows. Only two metabolites (i.e., sn-glycero-3-phosphocholine and phosphatidylethanolamine ae C42:1) were increased in the milk of lame cows. In conclusion, milk metabotyping of lame cows revealed significant changes in multiple milk components, including amino acids, lipids, and biogenic amines. Most of the milk compounds identified as altered were lowered, suggesting deflection of nutrients from the mammary gland to the host needs for healing lameness-associated pathological processes.

Colostrum-supplemented transition milk positively affects serum biochemical parameters, humoral immunity indicators and the growth performance of calves.

The aim of this study was to determine the effect of colostrum-supplemented transition milk on the growth rate of newborn calves. Thirty-six day-old female Holstein calves with a birth weight of 43.4 ± 5.5 kg were randomly assigned to a treatment group and a control group. On day 1, all calves received one portion (3 kg) of colostrum within 2 h after birth. Between the second feeding and the age of 5 days, control calves received only transition milk that had been collected from dams shortly before feeding, and the treatment group received 1.5 kg of colostrum collected from dams on day 1 and 1.5 kg of transition milk from subsequent milkings. Monthly body weight and average daily gain were used to measure growth performance. Blood samples were collected from the jugular vein at multiple time points to evaluate the biochemical and immune status of calves. Colostrum-supplemented transition milk increased the concentrations of serum immunoglobulins, triacylglycerols, cholesterol and total protein. The activity of lactate-dehydrogenase and ceruloplasmin decreased, whereas lysozyme activity increased with time in the control group. It can be concluded that additional five days of colostrum feeding positively influences non-specific humoral immunity indicators and serum biochemical parameters in dairy calves.

Mass-spec-based urinary metabotyping around parturition identifies screening biomarkers for subclinical mastitis in dairy cows.

The objective of this study was to determine urinary metabotypes of dairy cows prior to, during, and after diagnosis of subclinical mastitis (SCM). Twenty controls (CON) and 6 cows with SCM were included in the study. DI/LC-MS/MS was used to measure 186 metabolites in the urine at -8 and - 4 wks prepartum, disease diagnosis, and at +4 and + 8 wks postpartum. Results showed a total of 24 and 27 metabolites that differentiated SCM and CON cows at -8 and - 4 wks, respectively, with 5 top metabolites having an accuracy to predict SCM of 1.0, for both time point. Altered metabolites included several acylcarnitines (ACs), phosphatidylcholines (PCs), amino acids (AAs), and biogenic amines (BAs). During SCM diagnosis week there were a total of 22 metabolites that differentiated the SCM and CON cows including 13 ACs, 5 AAs, hexose, and phosphatidylethanolamine ae C44:3. The top 5 metabolites Symmetric dimethylarginine (SDMA), methylglutarylcarnitine, dodecanoylcarnitine, phosphatidylethanolamine ae C42:1, and phosphatidylethanolamine ae C42:0 showed an AUC of 1.0. Metabolite alterations continued at +4 and + 8 wks postpartum with 13 (9 ACs, 3 glycerophospholipids, and 1 BA) and 28 metabolites (14 ACs, 2 glycerophospholipids, hexose, 8 AAs, and 2 BAs) that differentiated the two groups, respectively. The top 5 most important metabolites, for both time points, showed AUCs of 1.0. Results of this study showed that typical urinary metabotypes preceded SCM event starting from -8 and - 4 wks prepartum. Metabotypes typical of SCM continued to be present during the week of SCM and at +4 and + 8 wks postpartum. These data show that the urine of dairy cows can be used with very high accuracy for screening dairy cows for susceptibility to SCM prior to entering into the dry off period and a potential new pen-side test can help dairy producers and veterinarians to make selective treatments.

Accuracy of registrations between cone-beam computed tomography and conventional computed tomography images and dose mapping methods in RaySearch software for the bladder during brachytherapy of cervical cancer patients.

PURPOSE: The aim of the study was to assess selected methods of image registration available in the RaySearch software and their impact on the accuracy of mapping of doses deposited in the bladder during brachytherapy (BRT) of cervical cancer in images used during external beam radiotherapy (EBRT). MATERIAL AND METHODS: The study was based on data from ten patients. Cone-beam computed tomography (CBCT) images (BRT) were aligned with CT images (EBRT) using four registration methods: Reg_1 (rigid), Reg_2a, Reg_2b (hybrid), and Reg_3 (biomechanical). Image mapping accuracy was evaluated based on bladder's anatomy. Sørensen-Dice coefficient (DSC) values were analyzed for all the registrations. Discrepancies between triangular mesh points set on the basis of bladder contours were analyzed. Dose distributions from BRT were transformed according to registration results and mapped on CT images. Original BRT doses deposited in 2 cm3 volume of the bladder were compared to those transformed and associated with bladder's volume determined on CT images. RESULTS: Mean DSC values amounted to 0.36 (Reg_1), 0.87 and 0.88 (Reg_2a and Reg_2b), and 0.97 (Reg_3). Significant differences were found between DSC for the following comparisons: Reg_3/Reg_1 (p = 0.001), Reg_2a/Reg_1 (p = 0.011), and Reg_2b/Reg_1 (p = 0.014). The lowest discrepancies between triangular mesh points were for Reg_3 (p < 0.001, Reg_3 vs. Reg_1, and p = 0.039, Reg_3 vs. Reg_2b). Finally, the lowest discrepancies between the original and transformed doses were found for Reg_3. Nevertheless, only 5 out of 10 observations for Reg_3 yielded error of less than 5%. CONCLUSIONS: Biomechanical registration (Reg_3) enabled the most accurate alignment between CBCT and CT images. Satisfactory registration results of anatomical structures do not guarantee a correct mapping of primary BRT doses on the bladder delineated on CT images during EBRT. The results of dose transformation based on biomechanical registration had an error of less than 5% for only 50% of the observations.

Serum metabolic fingerprinting of pre-lameness dairy cows by GC-MS reveals typical profiles that can identify susceptible cows.

The objectives of this study were to identify metabolite fingerprints in the serum related to amino acid (AA), carbohydrate, and lipid metabolism in transition dairy cows at -8 and -4 wks prior to parturition, at +2 wks postpartum during lameness diagnosis as well as at +4 and +8 wks after parturition. All cases of lameness occurred at around +2 wks after parturition. Out of 100 dairy cows included in this nested case-control study only 6 pregnant multiparous (parity: 3.0 ± 0.6, Mean ± SEM) Holstein dairy cows with lameness only and 20 healthy control cows (CON) were selected for serum GC-MS metabolomics analysis. All cows selected were not injured mechanically and had similar parity (3.3 ± 0.6) and body condition score (BCS). A total of 29 metabolites were identified and quantified in the serum. Results showed that 18 and 15 metabolites differentiated pre-lame cows from CON ones at -8 and -4 wks prior to parturition. Ten metabolites were found altered at the week of lameness diagnosis. Of note: pre-lame cows were characterized by greater concentrations of several amino acids including Gly, Leu, Phe, Ser, Val, D-mannose, Myo-inositol, and phosphoric acid (PA) at -8 and -4 wks prior to lameness and at the week of lameness diagnosis. At +4 wks after parturition 11 metabolites were altered in lameness cows, and at +8 wks there were 13 metabolites that differentiated the two groups. The high accuracy of the top 6 metabolites at -8 wks prior to parturition or approximately 9-11 wks before lameness diagnosis (Glu, Orn, Phe, Ser, Val, and PA) and another 5 metabolites at -4 wks before parturition, or approximately 5-7 wks before lameness diagnosis (Leu, Orn, Phe, Ser, and D-mannose) suggest that those metabolites may serve as potential monitoring biomarkers of lameness prior to lameness diagnosis. Data also showed multiple alterations during the week of lameness as well as at +4 and +8 wks postpartum suggesting lame cows are not metabolically healthy several weeks after the incidence of lameness. SIGNIFICANCE: Lameness is one of the top three health issues of dairy cows in Canada that influences early culling of dairy cows. Despite a few efforts, there is scarcity of data regarding metabolic alterations that precede, associate, and follow lameness. We investigated whether alterations in the metabolite signatures prior, during, and after development of lameness can be used to screen cows for susceptibility to lameness, characterize lameness from the metabolic prospective, and predict the outcome of this economically important health issue of dairy cows. The results demonstrate typical metabotypes as shown by increased serum concentrations of Val, Gly, Ser, Leu, Phe, D-mannose, myo-inositol, and phosphoric acid at -8 and -4 wks prior to parturition (or -6 to -10 wks prior to occurrence of lameness) and at the week of lameness diagnosis.