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2.
Res Vet Sci ; 172: 105240, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38608347

ABSTRACT

Antimicrobial usage (AMU) could be reduced by differentiating the causative bacteria in cases of clinical mastitis (CM) as either Gram-positive or Gram-negative bacteria or identifying whether the case is culture-negative (no growth, NG) mastitis. Immunoassays for biomarker analysis and a Tandem Mass Tag (TMT) proteomic investigation were employed to identify differences between samples of milk from cows with CM caused by different bacteria. A total of 94 milk samples were collected from cows diagnosed with CM across seven farms in Scotland, categorized by severity as mild (score 1), moderate (score 2), or severe (score 3). Bovine haptoglobin (Hp), milk amyloid A (MAA), C-reactive protein (CRP), lactoferrin (LF), α-lactalbumin (LA) and cathelicidin (CATHL) were significantly higher in milk from cows with CM, regardless of culture results, than in milk from healthy cows (all P-values <0.001). Milk cathelicidin (CATHL) was evaluated using a novel ELISA technique that utilises an antibody to a peptide sequence of SSEANLYRLLELD (aa49-61) common to CATHL 1-7 isoforms. A classification tree was fitted on the six biomarkers to predict Gram-positive bacteria within mastitis severity scores 1 or 2, revealing that compared to the rest of the samples, Gram-positive samples were associated with CRP < 9.5 µg/ml and LF ≥ 325 µg/ml and MAA < 16 µg/ml. Sensitivity of the tree model was 64%, the specificity was 91%, and the overall misclassification rate was 18%. The area under the ROC curve for this tree model was 0.836 (95% bootstrap confidence interval: 0.742; 0.917). TMT proteomic analysis revealed little difference between the groups in protein abundance when the three groups (Gram-positive, Gram-negative and no growth) were compared, however when each group was compared against the entirety of the remaining samples, 28 differentially abundant protein were identified including ß-lactoglobulin and ribonuclease. Whilst further research is required to draw together and refine a suitable biomarker panel and diagnostic algorithm for differentiating Gram- positive/negative and NG CM, these results have highlighted a potential panel and diagnostic decision tree. Host-derived milk biomarkers offer significant potential to refine and reduce AMU and circumvent the many challenges associated with microbiological culture, both within the lab and on the farm, while providing the added benefit of reducing turnaround time from 14 to 16 h of microbiological culture to just 15 min with a lateral flow device (LFD).


Subject(s)
Biomarkers , Mastitis, Bovine , Milk , Animals , Cattle , Female , Milk/chemistry , Milk/microbiology , Mastitis, Bovine/microbiology , Mastitis, Bovine/diagnosis , Biomarkers/metabolism , Proteome , Milk Proteins/analysis , Gram-Negative Bacteria/isolation & purification , Gram-Positive Bacteria/isolation & purification , Cathelicidins
3.
Rheumatology (Oxford) ; 63(1): 16-25, 2024 Jan 04.
Article in English | MEDLINE | ID: mdl-37647631

ABSTRACT

BACKGROUND: The aim of this systematic review and meta-analysis was to summarize current evidence regarding body composition (BC) in SSc in order to gain new insights and improve clinical care in the context of the nutritional status of SSc patients. METHODS: The databases Web of Science, PubMed, Scopus and Cochrane Library were searched on 4 January 2023. Studies were included if they provided data regarding BC obtained by dual-energy X-ray absorptiometry (DXA) or bioelectrical impedance analysis (BIA) in patients with SSc and healthy controls (HC). The study design criteria for inclusion were cohort and observational studies. The risk of bias assessment was performed using the Newcastle-Ottawa scale. For meta-analysis, mean difference with a 95% confidence interval was obtained and all results were depicted as forest plots. RESULTS: The number of retrieved publications was 593, of which nine were included in a random-effects meta-analysis totalling 489 SSc patients and 404 HC. Overall, significantly lower body mass index, lean mass (LM), fat mass (FM) and phase angle values were found in SSc patients when compared with HC. Furthermore, FM and LM were significantly lower in SSc patients when the DXA method was applied, whereas the same parameters were comparable between two groups of participants when BIA was applied. CONCLUSION: Altered BC is characteristic of SSc patients indicating the need for regular nutritional status assessment in order to improve the quality of life and clinical care of patients with SSc.


Subject(s)
Quality of Life , Scleroderma, Systemic , Humans , Body Composition , Nutritional Status , Absorptiometry, Photon/methods , Body Mass Index , Electric Impedance
4.
Diagnostics (Basel) ; 13(17)2023 Aug 26.
Article in English | MEDLINE | ID: mdl-37685307

ABSTRACT

Adropin is a secretory peptide that regulates glucose, lipid, and protein metabolism, which is closely related to obesity, insulin resistance, dyslipidemia, and atherogenesis. The serum adropin level is related to sex and depends upon nutritional preferences. This study aims to determine the association between serum adropin levels and body composition parameters in kidney transplant recipients (KTRs), especially emphasizing sex differences. Our case-control study involved 59 KTRs (28 postmenopausal women and 31 men) who were divided into two groups according to sex, and each group of those KTRs was further divided into higher or lower adropin values than the mean value in each sex group. Univariate regression showed a negative association of adropin levels with most anthropometric and body composition parameters in men's KTRs. Contrary to this, the serum adropin level was negatively associated only with phase angle in postmenopausal female KTRs. Multivariate regression showed that skeletal muscle mass and phase angle were the only negative predictors in women's KTRs, whereas in men, negative predictors were BMI and body water. These findings imply that adropin could have a different impact on metabolic homeostasis in KTRs regarding sex and could be considered a negative predictor of body composition in KTRs.

5.
J Pers Med ; 13(8)2023 Jul 28.
Article in English | MEDLINE | ID: mdl-37623450

ABSTRACT

Biologic disease-modifying antirheumatic drugs (DMARDs) are very effective in treating rheumatic diseases with a good patient tolerance. However, high cost and individualistic approach requires dedication of the physician. Therefore, the aim of this study was to determine how the COVID-19 pandemic has affected the prescription of biologic DMARDs in rheumatology at the University Hospital of Split. The data collection was conducted through an archive search in the Outpatient Clinic for Rheumatology in the University Hospital of Split, Split, Croatia. The search included the period before and after the start of the COVID-19 pandemic in Croatia (31 March 2020). Collected data included age, sex, ICD-10 code of diagnosis, generic and brand name of the prescribed drug, date of therapy initiation, and medication administration route. In the pre-COVID-19 period, 209 patients were processed, while in the COVID-19 period, 185 patients were processed (11.5% fewer). During pre-COVID-19, 231 biologic medications were prescribed, while during COVID-19, 204. During COVID-19, IL-6 inhibitors were less prescribed (48 (21%) vs. 21 (10%) prescriptions, p = 0.003), while IL-17A inhibitors were more prescribed (39 (17%) vs. 61 (30%) prescriptions, p = 0.001). In ankylosing spondylitis (AS), adalimumab was prescribed more during pre-COVID-19 (25 vs. 15 patients, p = 0.010), while ixekizumab was prescribed less (1 vs. 10 patients, p = 0.009). In rheumatoid arthritis, tocilizumab was prescribed more in the pre-COVID-19 period (34 vs. 10 patients, p = 0.012). Overall, the prescription trends of biologic DMARDs for rheumatologic diseases did not vary significantly in the University Hospital of Split, Croatia. Tocilizumab was prescribed less during COVID-19 due to shortages, while ixekizumab was more prescribed during COVID-19 due to an increase in psoriatic arthritis patients processed and due to being approved for treating AS.

6.
Nutrients ; 15(9)2023 Apr 24.
Article in English | MEDLINE | ID: mdl-37432203

ABSTRACT

The aim of this cross-sectional study was to determine the body fluid volume in patients diagnosed with both chronic kidney disease (CKD) and arterial hypertension (AH), and to investigate the relationship between fluid overload (FO), nutritional status and arterial stiffness in this specific patient population. A total of 169 participants with CKD and AH were enrolled in the study, and data on general parameters, comorbidities, medication use, and laboratory parameters were collected. Body composition was assessed with a Tanita MC 780 device, and data on the central and peripheral systolic and diastolic blood pressure, as well as pulse wave velocity (PWV) and the augmentation index (AIx) were collected with an IEM Mobil-O-Graph 24 h ambulatory blood pressure monitor, which was based on oscillometry. The Mediterranean Diet Serving Score (MDSS) questionnaire was used to determine the adherence to the Mediterranean diet (MeDi). Our results showed that the significant positive predictors of hydration status were the use of diuretics and oral hypoglycemic agents, whereas the negative predictors were female sex, higher body mass index level and use of two or more antihypertensives in the form of a single-pill combination. We also found differences in blood pressure and arterial stiffness parameters in relation to volume status, along with differences based on the presence of diabetes mellitus (DM). In conclusion, these results call for a higher awareness of volume status in the care of CKD patients with AH, especially in those with diabetes mellitus.


Subject(s)
Hypertension , Vascular Stiffness , Humans , Female , Male , Nutritional Status , Blood Pressure Monitoring, Ambulatory , Cross-Sectional Studies , Pulse Wave Analysis
7.
Sci Rep ; 13(1): 7475, 2023 05 08.
Article in English | MEDLINE | ID: mdl-37156785

ABSTRACT

The essential oils from the Centaurea genus are well known for their pharmacological properties. The most abundant and dominant chemical components in Centaurea essential oils are ß-caryophyllene, hexadecanoic acid, spathulenol, pentacosane, caryophyllene oxide, and phytol. However, whether these dominant components are the key drivers for observed antimicrobial activity remains unclear. Thus, the aim of this study was dual. Here we provide comprehensive, literature-based data to correlate the chemical compounds in Centaurea essential oils with the tested antimicrobial activity. Secondly, we characterized the essential oil of Centaurea triumfettii All. squarrose knapweed using coupled system gas chromatography-mass spectrometry and tested its phytochemicals for antimicrobial activity against E. coli and S. epidermis using disc diffusion assay and monitoring their growth in Muller Hinton broth. The most abundant compounds in C. triumfettii essential oil were hexadecanoic acid (11.1%), spathulenol (10.8%), longifolene (8.8%), germacrene D (8.4%), aromadendrene oxide (6.0%) and linoleic acid (5.3%). Based on our analysis of literature data from other Centaurea essential oils, they were positively correlated with antimicrobial activity. Using an agar disk diffusion method, tested chemical constituents did not show experimental evidence to support this positive correlation to antimicrobial activity when we tested them as pure components. The antibacterial effect of essential oil constituents may be related to a complex synergistic, rather than a single component as suggested by performed network pharmacology analysis, underlying the theoretical interactions between the essential oil phytochemicals listed as potentially responsible for antimicrobial activity and should be confirmed in further in-depth studies. This is the first report on the comparative analysis of Centaurea essential oils with good antimicrobial activity, as well as the first analysis of chemical components of the essential oil from C. triumfettii and the first report of antimicrobial activity of the representative, pure components: aromadendrene, germacrene D, spathulenol, longifolene, and the mixture of selected chemical compounds. This work contributes to the body of knowledge on the genus Centaurea and C. triumfettii species.


Subject(s)
Centaurea , Oils, Volatile , Oils, Volatile/chemistry , Centaurea/chemistry , Escherichia coli , Palmitic Acid , Anti-Bacterial Agents/pharmacology , Anti-Bacterial Agents/chemistry , Microbial Sensitivity Tests
8.
Diagnostics (Basel) ; 13(8)2023 Apr 16.
Article in English | MEDLINE | ID: mdl-37189530

ABSTRACT

BACKGROUND: The aim of the current systematic review was to summarize and evaluate the overall advantages of lung ultrasonography (LUS) examination using high-resolution computed tomography (HRCT) as a reference standard in assessing the presence of interstitial lung disease (ILD) in systemic sclerosis (SSc) patients. METHODS: Databases PubMed, Scopus, and Web of Science were searched for studies evaluating LUSs in ILD assessments including SSc patients on 1 February 2023. In assessing risk of bias and applicability, the Revised Tool for the Quality Assessment of Diagnostic Accuracy Studies (QUADAS-2) was used. A meta-analysis was performed and the mean specificity, sensitivity, and diagnostic odds ratio (DOR) with a 95% confidence interval (CI) were obtained. In addition, in a bivariate meta-analysis, the summary receiver operating characteristic (SROC) curve area was additionally calculated. RESULTS: Nine studies with a total of 888 participants entered the meta-analysis. A meta-analysis was also performed without one study that used pleural irregularity to assess the diagnostic accuracy of LUSs using B-lines (with a total of 868 participants). Overall sensitivity and specificity did not differ significantly, with only the analysis of the B-lines having a specificity of 0.61 (95% CI 0.44-0.85) and a sensitivity of 0.93 (95% CI 0.89-0.98). The diagnostic odds ratio of univariate analysis of the eight studies using the B-lines as a criterion for ILD diagnosis was 45.32 (95% CI 17.88-114.89). The AUC value of the SROC curve was 0.912 (and 0.917 in consideration of all nine studies), which indicates high sensitivity and a low false-positive rate for the majority of the included studies. CONCLUSIONS: LUS examination proved to be a valuable tool in discerning which SSc patients should receive additional HRCT scans to detect ILD and therefore reduces the doses of ionizing radiation exposure in SSc patients. However, further studies are needed to achieve consensus in scoring and the evaluation methodology of LUS examination.

9.
Semin Arthritis Rheum ; 60: 152200, 2023 06.
Article in English | MEDLINE | ID: mdl-37062151

ABSTRACT

BACKGROUND: This systematic review and meta-analysis aimed to summarize current evidence on vitamin D status in patients with psoriatic arthritis (PsA) with a particular focus on disease activity. METHODS: PubMed, Web of Science, Scopus and Cochrane Library databases were searched for studies that investigated vitamin D levels in PsA. The search was conducted on 12th October 2022. Included studies were cohorts, RCTs or observational studies, those assessing the level of 25(OH)D3 with control group consisting of healthy or psoriasis (Pso) patients. Nottingham-Ottawa Quality Scale was used to assess methodological quality. Random effects meta-analysis model was applied with inverse variance weighting and mean difference with 95% CI was calculated. RESULTS: Of 356 retrieved studies, 76 duplicates and 270 studies were excluded according to the exclusion criteria with one study unavailable. Four studies including 264 PsA patients and 287 healthy controls and five studies including 225 PsA patients and 391 Pso patients assessing vitamin D levels were eligible for meta-analysis. Vitamin D levels were lower in PsA patients compared to the healthy group (MD = -6.42; 95 % CI -8.31, -4.53; P < 0.01), while higher compared to Pso patients (MD = 2.37; 95 % CI 0.97, 3.78; P < 0.01). Included studies had moderate to low risk of bias. CONCLUSION: In conclusion, PsA patients have lower vitamin D levels than the general population. However, further studies are essential to understand the role of vitamin D in the development and treatment of PsA and the differences in vitamin D metabolism in PsA and Pso.


Subject(s)
Arthritis, Psoriatic , Psoriasis , Humans , Vitamin D , Risk
10.
Int J Mol Sci ; 24(8)2023 Apr 10.
Article in English | MEDLINE | ID: mdl-37108169

ABSTRACT

Infectious diseases still threaten global human health, and host genetic factors have been indicated as determining risk factors for observed variations in disease susceptibility, severity, and outcome. We performed a genome-wide meta-analysis on 4624 subjects from the 10,001 Dalmatians cohort, with 14 infection-related traits. Despite a rather small number of cases in some instances, we detected 29 infection-related genetic associations, mostly belonging to rare variants. Notably, the list included the genes CD28, INPP5D, ITPKB, MACROD2, and RSF1, all of which have known roles in the immune response. Expanding our knowledge on rare variants could contribute to the development of genetic panels that could assist in predicting an individual's life-long susceptibility to major infectious diseases. In addition, longitudinal biobanks are an interesting source of information for identifying the host genetic variants involved in infectious disease susceptibility and severity. Since infectious diseases continue to act as a selective pressure on our genomes, there is a constant need for a large consortium of biobanks with access to genetic and environmental data to further elucidate the complex mechanisms behind host-pathogen interactions and infectious disease susceptibility.


Subject(s)
Communicable Diseases , Genetic Predisposition to Disease , Humans , Phenotype , Risk Factors , Genome-Wide Association Study , Communicable Diseases/genetics , Nuclear Proteins/genetics , Trans-Activators/genetics
11.
Nat Hum Behav ; 7(5): 790-801, 2023 05.
Article in English | MEDLINE | ID: mdl-36864135

ABSTRACT

Identifying genetic determinants of reproductive success may highlight mechanisms underlying fertility and identify alleles under present-day selection. Using data in 785,604 individuals of European ancestry, we identified 43 genomic loci associated with either number of children ever born (NEB) or childlessness. These loci span diverse aspects of reproductive biology, including puberty timing, age at first birth, sex hormone regulation, endometriosis and age at menopause. Missense variants in ARHGAP27 were associated with higher NEB but shorter reproductive lifespan, suggesting a trade-off at this locus between reproductive ageing and intensity. Other genes implicated by coding variants include PIK3IP1, ZFP82 and LRP4, and our results suggest a new role for the melanocortin 1 receptor (MC1R) in reproductive biology. As NEB is one component of evolutionary fitness, our identified associations indicate loci under present-day natural selection. Integration with data from historical selection scans highlighted an allele in the FADS1/2 gene locus that has been under selection for thousands of years and remains so today. Collectively, our findings demonstrate that a broad range of biological mechanisms contribute to reproductive success.


Subject(s)
Fertility , Reproduction , Child , Female , Humans , Aging/physiology , Fertility/genetics , Menopause/genetics , Reproduction/genetics , Selection, Genetic
12.
Nutrients ; 15(3)2023 Jan 18.
Article in English | MEDLINE | ID: mdl-36771203

ABSTRACT

BACKGROUND: The aim of the present review was to summarize the current evidence about the impact of vitamin D deficiency on pathology and clinical manifestations of Sjögren's disease (SD). METHODS: Databases PubMed, Web of Science, Scopus, and Cochrane library were searched for studies assessing the levels of vitamin D in SD patients using the following keywords: (vitamin D OR calciferol OR cholecalciferol OR 25-hydroxyvitamin D OR 25-hydroxycholecalciferol OR calcidiol OR calcitriol OR 1,25-dihydroxycholecalciferol) AND (Sjögren's Syndrome OR Sjögren's disease) accessed on 20 September 2022. Out of 248 retrieved studies, following the systematic review methodology and defined inclusion and exclusion criteria, 9 clinical studies were eligible to be included in the present review: 4 of them case-control, 4 cross-sectional, and 1 cohort study. RESULTS: Nine studies totaling 670 SD patients and 857 healthy controls were eligible for meta-analysis with moderate to high methodological quality as determined by the Newcastle-Ottawa Quality Scale (NOS). According to the obtained results, a high prevalence of hypovitaminosis D was observed in SD patients when compared to healthy controls (95% CI -10.43, -2.39; p < 0.01). CONCLUSION: Available evidence points to lower levels of vitamin D in patients with SD in comparison to healthy controls. However, further studies are necessary to understand the underlying mechanisms associated with the role of vitamin D in the development and disease severity of SD.


Subject(s)
Sjogren's Syndrome , Vitamin D Deficiency , Humans , Sjogren's Syndrome/epidemiology , Sjogren's Syndrome/complications , Cohort Studies , Cross-Sectional Studies , Vitamin D , Vitamins , Calcifediol , Vitamin D Deficiency/complications , Vitamin D Deficiency/epidemiology
13.
Brain ; 146(7): 3049-3062, 2023 07 03.
Article in English | MEDLINE | ID: mdl-36730021

ABSTRACT

Personalized management of neuropathic pain is an unmet clinical need due to heterogeneity of the underlying aetiologies, incompletely understood pathophysiological mechanisms and limited efficacy of existing treatments. Recent studies on microRNA in pain preclinical models have begun to yield insights into pain-related mechanisms, identifying nociception-related species differences and pinpointing potential drug candidates. With the aim of bridging the translational gap towards the clinic, we generated a human pain-related integrative miRNA and mRNA molecular profile of the epidermis, the tissue hosting small nerve fibres, in a deeply phenotyped cohort of patients with sodium channel-related painful neuropathy not responding to currently available therapies. We identified four miRNAs strongly discriminating patients from healthy individuals, confirming their effect on differentially expressed gene targets driving peripheral sensory transduction, transmission, modulation and post-transcriptional modifications, with strong effects on gene targets including NEDD4. We identified a complex epidermal miRNA-mRNA network based on tissue-specific experimental data suggesting a cross-talk between epidermal cells and axons in neuropathy pain. Using immunofluorescence assay and confocal microscopy, we observed that Nav1.7 signal intensity in keratinocytes strongly inversely correlated with NEDD4 expression that was downregulated by miR-30 family, suggesting post-transcriptional fine tuning of pain-related protein expression. Our targeted molecular profiling advances the understanding of specific neuropathic pain fine signatures and may accelerate process towards personalized medicine in patients with neuropathic pain.


Subject(s)
MicroRNAs , Neuralgia , Humans , RNA, Messenger , Neuralgia/genetics , Neuralgia/metabolism , Epidermis/metabolism , MicroRNAs/genetics , Epidermal Cells/metabolism , NAV1.7 Voltage-Gated Sodium Channel/genetics , NAV1.7 Voltage-Gated Sodium Channel/metabolism
14.
Prep Biochem Biotechnol ; 53(9): 1067-1080, 2023 Oct.
Article in English | MEDLINE | ID: mdl-36645251

ABSTRACT

Histones are an essential part of nucleosomes that regulate chromatin structure and function. Histone exchanges and modifications represent a scaffold for DNA transcription, repair, and replication. Studying histones and histone code is an important and fast-developing branch of epigenetic science. Here we propose a fast, efficient, and versatile assay for nucleosomal histone isolation from mammalian cells, without the use of acids or high salt solutions which are common for other histone isolation techniques. All components used in the protocol are common and inexpensive laboratory chemicals. The protocol has been evaluated on six commonly used cell lines and two animal tissue samples. The mild extraction conditions preserve delicate histone epigenetic changes, allowing its downstream analyses. We have demonstrated the assays' successful application during changes in the transcriptional activity of histone genes, cell cycle transitions, and DNA-damaging conditions. Histone fractions, obtained by the protocol, can be used for further applications, such as electrophoresis, immunoblot, and mass spectrometry. Therefore, the new proposed nucleosomal histone isolation method is sensitive, specific, and suitable for downstream applications of various kinds.


Subject(s)
Histones , Nucleosomes , Animals , Histones/chemistry , Histones/genetics , Histones/metabolism , Cost-Benefit Analysis , DNA , Cell Cycle , Mammals/genetics , Mammals/metabolism
15.
J Periodontol ; 94(6): 765-776, 2023 06.
Article in English | MEDLINE | ID: mdl-36700464

ABSTRACT

BACKGROUND: Kidney transplant recipients (KTRs) represent a vulnerable group of patients who develop a number of comorbidities. Severe periodontitis (SP) is associated with the most common chronic systemic diseases including kidney diseases. The objective of this study was to explore the risk factors for SP in KTRs. METHODS: In this study, KTRs were divided into those with or without periodontitis and in relation to the severity of periodontitis. A comprehensive medical and periodontal examination was performed and evaluated. Multivariate logistic regression was performed to examine possible risk factors for SP among KTRs. RESULTS: A total of 100 KTRs were included in the analysis, of which 87% had periodontitis. Significant predictors of periodontitis were older age (OR = 1.07, 95% CI [1.01, 1.13], p = 0.016) and lower skeletal muscle mass (OR = 0.88, 95% CI [0.78, 0.99], p = 0.035). When examining periodontitis severity, predictors of SP (n = 21, 24%) were increased levels of uric acid (OR = 1.01, 95% CI [1.00, 1.02], p = 0.022) and dental plaque (OR = 1.04, 95% CI [1.01, 1.07], p = 0.013). In the subset analysis that included only KTRs with measured advanced glycation end products (AGE) (n = 47), 34% (n = 16) had SP. The predictors of SP were AGE (OR = 3.89, 95% CI [1.28, 11.82], p = 0.017) and dental plaque (OR = 1.07, 95% CI [1.01, 1.13], p = 0.028). CONCLUSIONS: KTRs with SP had significantly higher uric acid levels and AGE, which may contribute to the systemic health status of this patient population.


Subject(s)
Dental Plaque , Kidney Transplantation , Periodontitis , Humans , Case-Control Studies , Kidney Transplantation/adverse effects , Uric Acid , Risk Factors , Periodontitis/complications , Periodontitis/epidemiology
16.
Nucleic Acids Res ; 51(6): e35, 2023 04 11.
Article in English | MEDLINE | ID: mdl-36718861

ABSTRACT

DNA-protein crosslinks (DPCs), formed by the covalent conjugation of proteins to DNA, are toxic lesions that interfere with DNA metabolic processing and transcription. The development of an accurate biochemical assay for DPC isolation is a priority for the mechanistic understanding of their repair. Here, we propose the STAR assay for the direct quantification of DPCs, sensitive to physiologically relevant treatment conditions. Implementing the STAR assay revealed the formation of small cross-linked peptides on DNA, created by the proteolytic degradation of DPCs by SPRTN. The initial proteolytic degradation of DPCs is required for the downstream activation of DNA repair, which is mediated through the phosphorylation of H2Ax. This leads to the accumulation of DNA repair factors on chromatin and the subsequent complete removal of the cross-linked peptides. These results confirmed that the repair of DPCs is a two-step process, starting with proteolytic resection by SPRTN, followed by the repair of the underlying damage to the DNA.


Subject(s)
DNA Damage , DNA-Binding Proteins , DNA-Binding Proteins/genetics , DNA/genetics , DNA/metabolism , DNA Repair , Proteolysis , Peptide Hydrolases/genetics
17.
J Proteomics ; 270: 104735, 2023 01 06.
Article in English | MEDLINE | ID: mdl-36174949

ABSTRACT

Canine babesiosis is a tick-borne disease caused by Babesia canis, with acute kidney injury as one of the common complications. In the study 8 healthy control dogs and 22 dogs with naturally occurring babesiosis were enrolled, with the aim to analyse differences in serum and urinary proteomes between healthy dogs and dogs with different degree of renal dysfunction in babesiosis using a label-based high-throughput quantitative proteomic approach. In serum, 58 proteins were found differentially abundant between healthy controls and groups of dogs with different degrees of renal dysfunction in babesiosis, while in urine there were 259 differentially abundant proteins. In addition, altered biological pathways were detected in the diseased dogs using bioinformatics tools and validation of several candidate biomarkers was performed. SIGNIFICANCE: The main aim of this comprehensive study was to perform analyses of serum and urinary proteomes of dogs with renal dysfunction in babesiosis compared to healthy dogs using, for the first time, a high-throughput proteomic method and functional enrichment analyses. Serum and urine samples of the same dogs were investigated in order to gain a more complete picture of pathologic changes taking place in renal dysfunction in babesiosis. We highlighted two putative biomarkers validated herein which could be of importance for early diagnosis of renal dysfunction in canine babesiosis, as they are easily accessible from urine and their concentration rises before the appearance of azotaemia: urinary neutrophil gelatinase-associated lipocalin (NGAL) and urinary liver-type fatty acid-binding protein (L-FABP).


Subject(s)
Babesiosis , Dog Diseases , Kidney Diseases , Dogs , Animals , Babesiosis/complications , Babesiosis/diagnosis , Proteomics , Proteome , Dog Diseases/diagnosis , Biomarkers , Kidney Diseases/diagnosis , Kidney Diseases/urine , Kidney Diseases/veterinary
18.
J Proteomics ; 270: 104739, 2023 01 06.
Article in English | MEDLINE | ID: mdl-36174954

ABSTRACT

This study evaluates how long-term dietary low ω6:ω3 ratio in sows and offspring's seaweed (SW) intake affects piglet intestinal function and growth through modifying ileum proteome. Sows were assigned to either control diet (CR, ω6:ω3 ratio = 13:1) or treatment diet (LR, ω6:ω3 = 4:1) during gestation and lactation (n = 8 each). The male weaned offspring were received a basal diet with or without SW powder supplementation (4 g/kg) for 21 days, denoted as SW and CT groups, respectively. In total, four groups of weaned piglets were formed following maternal and offspring's diets combination, represented by CRCT, CRSW, LRCT, and LRSW (n = 10 each). Piglet ileum tissue was collected on day 22 post-weaning and analysed using TMT-based quantitative proteomics. The differentially abundant proteins (n = 300) showed the influence of maternal LR diet on protein synthesis, cell proliferation, and cell cycle regulation. In contrast, the SW diet lowered the inflammation severity and promoted ileal tissue development in CRSW piglets but reduced the fat absorption capacity in LRSW piglets. These results uncovered the mechanism behind the anti-inflammation and intestinal-boosting effects of maternal LR diet in piglets supplemented with SW.


Subject(s)
Proteome , Seaweed , Swine , Animals , Male , Female , Proteomics , Diet , Dietary Supplements , Lactation , Ileum , Vegetables , Animal Feed/analysis
19.
J Proteomics ; 270: 104740, 2023 01 06.
Article in English | MEDLINE | ID: mdl-36191802

ABSTRACT

This study examines whether maternal low ω6:ω3 ratio diet and offspring SW supplementation can improve offspring immunity and performance by elucidating the effects on piglet serum proteome. A total of 16 sows were given either a standard (CR, 13:1) or low ω6:ω3 ratio diet (LR, 4:1) during pregnancy and lactation and their male weaned piglets were supplemented with SW powder (4 g/kg, SW) or not (CT) in a 21-day post-weaning (PW) diet. Four PW piglet groups were then identified based on dam and piglet treatment, namely CRCT, CRSW, LRCT, and LRSW (n = 10 each). Piglet serum collected at weaning and d21 PW were analysed (n = 5 each) using TMT-based quantitative proteomics and validated by appropriate assays. The differentially abundant proteins (n = 122) displayed positive effects of maternal LR diet on anti-inflammatory properties and innate immune stimulation. Progeny SW diet activated the innate immunity and enhance the host defence during inflammation. These data demonstrate the value of decreasing ω6:ω3 ratio in maternal diet and SW supplementation in PW piglet's diet to boost their immunity and anti-inflammation properties. SIGNIFICANCE: This novel proteomic study in post-weaned piglets addresses the interplay between maternal and offspring nutritional interventions in a context of rapid and dynamic alterations in piglet metabolic status around weaning. Decreasing ω6:ω3 ratio in maternal diet and SW supplementation in PW piglet's diet can boost their immunity and anti-inflammation properties. This study also provides new insights into piglet serum proteome regulation during post-weaning, a critical development period in swine.


Subject(s)
Seaweed , Pregnancy , Swine , Animals , Female , Male , Proteome , Proteomics , Diet , Dietary Supplements , Vegetables , Animal Feed/analysis
20.
Article in English | MEDLINE | ID: mdl-36078776

ABSTRACT

There is limited evidence on the associations between dietary patterns, body composition, and nonclassical predictors of worse outcomes such as advanced glycation end products (AGE) in kidney transplant recipients (KTRs). The aim of this cross-sectional study was to determine the level of AGE-determined cardiovascular (CV) risk in Dalmatian KTRs and possible associations between AGE, adherence to the Mediterranean diet (MeDi), and nutritional status. Eighty-five (85) KTRs were enrolled in this study. For each study participant, data were collected on the level of AGE, as measured by skin autofluorescence (SAF), Mediterranean Diet Serving Score (MDSS), body mass composition, anthropometric parameters, and clinical and laboratory parameters. Only 11.76% of the participants were adherent to the MeDi. Sixty-nine percent (69%) of KTRs had severe CV risk based on AGE, while 31% of KTRs had mild to moderate CV risk. The results of the LASSO regression analysis showed that age, dialysis type, dialysis vintage, presence of CV and chronic kidney disease, C- reactive protein level, urate level, percentage of muscle mass, and adherence to recommendations for nuts, meat, and sweets were identified as positive predictors of AGE. The negative predictors for AGE were calcium, phosphate, cereal adherence according to the MeDi, and trunk fat mass. These results demonstrate extremely low adherence to the MeDi and high AGE levels related CV risk in Dalmatian KTRs. Lifestyle interventions in terms of CV risk management and adherence to the MeDi of KTRs should be taken into consideration when taking care of this patient population.


Subject(s)
Diet, Mediterranean , Glycation End Products, Advanced , Transplant Recipients , Body Composition , Cross-Sectional Studies , Glycation End Products, Advanced/metabolism , Humans , Kidney Transplantation
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