Evaluation of circulating microRNA profiles in blood as potential candidate biomarkers in a subacute ruminal acidosis cow model - a pilot study.

BACKGROUND: Subacute ruminal acidosis (SARA) is a metabolic disorder often observed in high-yielding dairy cows, that are fed diets high in concentrates. We hypothesized that circulating miRNAs in blood of cows could serve as potential candidate biomarkers to detect animals with metabolic dysbalances such as SARA. MicroRNAs (miRNAs) are a class of small non-coding RNAs, serving as regulators of a plethora of molecular processes. To test our hypothesis, we performed a pilot study with non-lactating Holstein-Friesian cows fed a forage diet (FD; 0% concentrate, n = 4) or a high-grain diet (HG; 65% concentrate, n = 4) to induce SARA. Comprehensive profiling of miRNA expression in plasma and leucocytes were performed by next generation sequencing (NGS). The success of our model to induce SARA was evaluated based on ruminal pH and was evidenced by increased time spent with a pH threshold of 5.8 for an average period of 320 min/d. RESULTS: A total of 520 and 730 miRNAs were found in plasma and leucocytes, respectively. From these, 498 miRNAs were shared by both plasma and leucocytes, with 22 miRNAs expressed exclusively in plasma and 232 miRNAs expressed exclusively in leucocytes. Differential expression analysis revealed 10 miRNAs that were up-regulated and 2 that were down-regulated in plasma of cows when fed the HG diet. A total of 63 circulating miRNAs were detected exclusively in the plasma of cows with SARA, indicating that these animals exhibited a higher number and diversity of circulating miRNAs. Considering the total read counts of miRNAs expressed when fed the HG diet, differentially expressed miRNAs ( log2 fold change) and known function, we have identified bta-miR-11982, bta-miR-1388-5p, bta-miR-12034, bta-miR-2285u, and bta-miR-30b-3p as potential candidates for SARA-biomarker in cows by NGS. These were further subjected to validation using small RNA RT-qPCR, confirming the promising role of bta-miR-30b-3p and bta-miR-2285. CONCLUSION: Our data demonstrate that dietary change impacts the release and expression of miRNAs in systemic circulation, which may modulate post-transcriptional gene expression in cows undergoing SARA. Particularly, bta-miR-30b-3p and bta-miR-2285 might serve as promising candidate biomarker predictive for SARA and should be further validated in larger cohorts.

Feeding dairy cows bakery by-products enhanced nutrient digestibility, but affected fecal microbial composition and pH in a dose-dependent manner.

We reported recently that adding bakery by-products (BP) to the diets of dairy cows up to 30% improved performance and rumen pH, but caused major shifts in the nutrient profile and availability, likely modifying nutrient degradation patterns throughout the gastrointestinal tract. The aim of this study was to investigate the effects of the gradual replacement of cereals by BP on the apparent total-tract digestibility (ATTD), the fermentation patterns, and the microbial community in feces of dairy cows. Twenty-four mid-lactating Simmental cows (149 ± 22.3 days in milk, 756 ± 89.6 kg of initial body weight) were fed a total mixed ration ad libitum (fresh feed was offered twice per day) containing a 50:50 ratio of forage to concentrate (dry matter basis) throughout the experiment. The trial lasted 5 wk, whereby the first week was used for baseline measurements, in which all cows received the same diet, without BP. Cows were then randomly allocated into 3 groups differing in the BP content of diets (0% BP, 15% BP, and 30% BP on a DM basis) and fed for 4 wk. Fecal samples were taken for analysis of pH, volatile fatty acids (VFA), and 16S rRNA gene sequencing. The inclusion of BP resulted in an increase of ether extract and sugars, and a reduction of starch and neutral detergent fiber in the diet. Feeding BP linearly increased the ATTD of almost all nutrients resulting in up to 2 kg more digestible organic matter intake (DOMI). Increasing BP level up to 30% increased fecal total VFA concentration and decreased the pH. The proportion of butyrate in feces increased linearly, but the proportion of all other VFA was not affected by BP-feeding. The richness and diversity indices of the fecal microbiota linearly declined by the inclusion of BP. The cellulolytic phyla Fibrobacteres decreased, whereas amylolytic phyla, such as Proteobacteria, increased. Overall, results showed that feeding BP linearly increased ATTD and DOMI, but impaired fecal microbial diversity and pH. In the interest of the optimization of BP inclusion in the dairy cows' feeding, a dietary level between 15 to 30% of BP might be a better compromise than 30% in terms of an enhanced DOMI and performance with still lowered risk of hindgut dysbiosis, but this will require further investigations.

Distinct serum metabolomic signatures of multiparous and primiparous dairy cows switched from a moderate to high-grain diet during early lactation.

INTRODUCTION: Feeding of high-grain diets is common in cows during early lactation, but increases the odds of metabolic derailments, which can likely be detected as undesirable shifts in the serum metabolome signature. OBJECTIVES: The present study aimed to identify the metabolic signatures of the serum metabolome of early lactation dairy cows switched from a moderate to a high-grain diet. METHODS: Targeted ESI-LC-MS/MS-based metabolomics was used to characterize metabolic alterations in the serum of early lactation multiparous (MP, n = 16) and primiparous (PP, n = 8) Simmental cows, according to parity and feeding phase. Data were analysed using different data mining approaches. RESULTS: Carnitine, acetylcarnitine, propionoylcarnitine, amino acid related compounds cis-4-hydroxyproline, trans-4-hydroxyproline, proline betaine, lysophosphatidylcholine PC a C16:1 and phosphatidylcholine PC ae C36:0 were identified as the key metabolites distinguishing MP from PP cows. A different serum metabolite composition during moderate and high-grain diet was also evident. Notably, cows fed high grain diet had higher serum concentrations of primary bile acids and triglycerides, but lower levels of conjugated bile acids and carboxylic acids during the first week in grain. Amino acids valine, cystine and taurine together with lysophosphatidylcholine PC a C26:0 and several phosphatidylcholines were classified as important features for cluster separation. CONCLUSIONS: Our study greatly expands earlier observations on dietary effects on serum metabolome composition of cows. The altered metabolomic fingerprints clearly distinguishable by diet and cow parity hold potential to be used as early diagnostic tools for cows experiencing grain-induced metabolic disturbances.

First experience in Italy with a new transcutaneous bone conduction implant.

Since 2011, transcutaneous bone-anchored auditory implants have been an alternative to the classic percutaneous implant (Baha) for bilateral conductive/mixed hearing loss that cannot be corrected by surgery. Recently, a new transcutaneous device has been approved for clinical use. Its internal component is made of the classic titanium Baha fixture, coupled to a 27 mm diameter subcutaneous circular magnet. The external component includes a second circular magnet 29 mm in diameter and a digital sound processor. To date, there are no reports describing the results of the application of this device. The aim of the present study is to report on the anatomical and functional results of transcutaneous Baha implantation in three patients: two adults, one with syndromic aural atresia and one with bilateral conductive hearing loss due to bilateral tympanomastoidectomy, and an 8-year-old child with non-syndromic aural atresia. No major intraoperative or postoperative complications were observed. The three patients tolerated the external magnet, with no signs of skin irritation. Functional results were good: median unaided free-field PTA (0.5-3 kHz) was 50 dB HL (range = 41-66 dB HL); with the transcutaneous Baha median PTA (0.5-3 kHz) was 27 dB HL (range = 25-30 dB HL) and median gain was 25 dB HL (range = 11-39 dB HL). Preliminary results encourage use of the device as a valuable alternative to other implantable devices in these patients. To ensure the success of treatment, several precautions are suggested including gradually increasing use during the first post-operative months to favour skin adaptation to magnet pressure. In addition to skin reactions, in a paediatric age most concerns are related to the curvature of the skull, which may induce tenting of the skin over the internal magnet.

Audiological findings in patients with microdeletion 22q11 (di George/velocardiofacial syndrome).

Microdeletion 22q11 (del22q11) is one of the most frequent causes of genetic syndromes. The majority of cases of di George and velocardiofacial syndromes are due to del22q11. These conditions are considered to be developmentally related to neural crest anomalies influencing the differentiation of the branchial arches, including the percursor tissue of the ear. In addition, the UFDIL gene, an ubiquination gene being expressed during embryogenesis in the inner ear primordia, has been identified in the 22q11 critical region. The aim of this study was to evaluate the prevalence of hearing impairment in del22q11 syndrome. Admittance audiometry, behavioural pure tone audiometry and auditory brainstem response (ABR) were performed in 27 children studied at our hospital between 1997 and 1998. Results were related to clinical history, frequency otitis media and immune status. Sensorineural hearing loss was found in 4/27 (15%) patients (severe in three cases, mild in one), conductive hearing impairment in 12/27 (45%) (moderate in four cases, mild in eight) and normal hearing in 11/27 (40%). Interestingly, three of the patients with sensorineural hearing loss had cerebral lesions due to neonatal distress, to hydrocephalus and to post-surgical ischaemia each in one. The prevalence of speech delay, otitis media and low CD3 values was higher among patients with conductive hearing impairment in comparison with those with normal hearing. In conclusion, hearing impairment was documented in 60% of the patients and must be included among the clinical features of del22q11 syndrome. Audiological evaluation is recommended in patients with del22q11 in order to reduce the risk of speech deficit.

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry of DNA-Pt(II) complexes.

Matrix-assisted laser desorption ionization (MALDI) time-of-flight mass spectrometry has been used to characterize the reaction products of the 18-mer deoxyribonucleotide d(AACGGTTAACCGTTAATT) with [Pt(NH3)3(H2O)]2+ and cis-[Pt(NH3)2(H2O)2]2+. Characteristic peaks corresponding to different monofunctional adducts (18-mer+n[Pt(NH3)3]) (n = 1, 2, 3 and 4) have been observed with the triamino-monoaqua complex. With the diamino-diaqua cis-Pt complex, formation of a chelate (18-mer+[Pt(NH3)2]) involving two adjacent guanines has been demonstrated. A good correlation between MALDI and polyacrylamide gel electrophoresis results is observed.

Attitudes toward war in adolescents.

180 college students did not differ from 31 high school students in positive attitudes toward war or cynicism. College students held, however, less patriotic attitudes.