The dysfunction of immune system and inflammation contribute to the Parkinson’s disease (PD) pathogenesis. Cytokines, oxidative stress, neurotoxin and metabolism associated enzymes participate in neuroinflammation in PD and the genes involved in them have been reported to be associated with the risk of PD. In our study, we performed a quantitative and causal analysis of the relationship between inflammatory genes and PD risk.Standard process was performed for quantitative analysis. Allele model (AM) was used as primary outcome analysis and dominant model (DM) and recessive model (RM) were applied to do the secondary analysis. Then, for those genes significantly associated with the risk of PD, we used the published GWAS summary statistics for Mendelian Randomization (MR) to test the causal analysis between them.IL-6 rs1800795, TNF-α rs1799964, PON1 rs854560, CYP2D6 rs3892097, HLA-DRB rs660895, BST1 rs11931532, CCDC62 rs12817488 polymorphisms were associated with the risk of PD statistically with the ORs ranged from 0.66 to 3.19 while variants in IL-1α, IL-1β, IL-10, MnSOD, NFE2L2, CYP2E1, NOS1, NAT2, ABCB1, HFE and MTHFR were not related to the risk of PD. Besides, we observed that increasing ADP-ribosyl cyclase (coded by BST1) had causal effect on higher PD risk (OR[95%CI] =1.16[1.10-1.22]) while PON1(coded by PON1) shown probably protective effect on PD risk (OR[95%CI] =0.81[0.66-0.99]).We included 36 variants in 18 genes for final pooled analysis. As a result, IL-6, TNF-α, PON1, CYP2D6, HLA-DRB, BST1, CCDC62 were statistically associated with the susceptibility of PD, and with evidence of causal relationships for ADP-ribosyl cyclase and PON1 on PD risk, which may help understand the mechanisms and pathways underlying PD pathogenesis.Several polymorphisms from inflammatory genes of  Its typvia several pathways  is a reliable genetic epidemiology method, which uses genetic variants as instrumental variable (IV) to assure whether causality exists between exposure and outcome, maybe a powerful tool to explore the causality between inflammation and PD . BottiglTherefore, to reach a comprehensive and updated conclusion, we performed a quantitative and causal analysis to explore the role of inflammatory genes in PD risk in order to bring new understanding of the mechanisms and pathways underlying PD pathogenesis and may provide the theoretical basis for finding the potential biomarkers and implementing anti-inflammatory and immunological treatment in PD. In addition to the genes included in the existing studies, we collected the original researches related to inflammation-related genes and PD as much as possible. Based on the function of genes, we divided them into five groups: genes of cytokines, genes involved in the oxidative stress, genes of neurotoxin-associated enzymes, genes of metabolism-associated enzymes and inflammatory polymorphic locus identified by GWAS study Figure 122.12.1.1TNF-α, IL-6, IL-1α, IL-1β, IL-10, NOS1, MnSOD, NFE2L2, CYP2D6, PON1, CYP2E1, NAT2, ABCB1, BST1, HLA-DRB, CCDC62, HFE, MTHFR involved in five different inflammation-related group  in PD). The detailed searching strategy was listed in Researchers independently retrieved and screened literature, and the inconsistent views were discussed with the third party. Key words were “Parkinson’s disease”, “Parkinso*”, “variants”, “genetic”, “specific genes”  were applied to screen articles:Participants: the PD diagnosis from each researched cohort was according to widely accepted criteria , 37.Interventions: genetic sequencing of variants in inflammation-related genes of interest were performed by PCR-based methods or other accepted methods;Controls: controls were neither having PD nor other neurological diseases.Outcomes: available data to calculate the number of carriers and non-carriers of the gene variants.Studies approach: original studies provided sufficient data to do pooled analysis.Exclusion criteria including: 1) neurological diseases not PD or without control groups; 2) not original studies including editorial, review, systematic review etc.; 3) functional studies using animal or cell models; 4) studies not having sufficient data to calculate odd ratio (OR) and 95% confidence interval (CI) in all models.2.1.3Then, authors independently extracted the detailed information from the included studies. The data extraction table were as follows: first author, publication year, ethnics, number of allele carriers in cases or controls, number of cases, number of controls, number of genotype carriers in cases, number of genotypes in controls. The Newcastle-Ottawa Scale (NOS) scores were used to evaluate the quality of the included articles. If there was any disagreement on data extraction, a third researcher was asked to make a decision.2.1.4Revman 5.3 software was used to calculate pooled OR and 95%CI. Three models were applied to do the association analyses: allele model , dominant model , and recessive model . “A” represented wild type allele, “a” represented mutated allele. P <0.05 was considered statistically significant.2 and Q test were performed to analyze the heterogeneity. If I2> 50, the random-effect model was used, otherwise if I2 ≤ 50, the fix-effect model was applied. The publication bias was measured by the symmetry of funnel plot. If the plot was in a symmetrical shape, no publication bias was shown. Otherwise, publication bias was observed. Sensitivity analysis was performed by sequentially removing one article at a time.The I2.22.2.1BST1) and PON1 (coded by PON1)] and PD in two directions  Study that contained 482,730 individuals with 37,688 PD from Europe . ADP-rib2.2.3p < 5 × 10−8  selection in two-sample MR analysis: (1) the selected genetic variants are associated with the exposure; (2) the used IV variants are not associated with the confounders of exposure and outcome; (3) there is no feasible pathway between the genetic variations and outcome other than through exposure . In deta5 × 10−8 –40. Then2.2.4We used the method of inverse-variance weighting (IVW)  and Mend33.1As can be seen from the flowchart  were included in the pooled analysis. In the results of DM and RM models, rs1799964 of TNF-a  polymorphism was positively associated with PD risk. In contrary, rs1800795 of IL-6  polymorphism was negatively associated with PD risk. About variants in IL-1α (rs1800587), IL-1β(rs16944), IL-10 , all three models  showed these variants were not associated with the risk of PD.Seven variants in five genes  were included for quantitative analysis. We failed to identify the association between NFE2L2 rs6706649, rs6721961, rs35652124, rs2706110, rs10183914, rs1806649, rs2001350, NOS1 rs2682826, rs1060826, MnSOD rs4880 and PD risk in all three models .Ten variants in three genes  were included in the AM for quantitative analysis. CYP2D6 rs3892097  variant was positively associated with PD risk on 1727 PD cases and 2087 controls. PON1 rs854560  variant was also positively associated with PD risk in AM on 2781 PD cases and 3176 controls.Eleven variants in five genes . The significant result was also replicated by DM in CYP2D6 variant rs3892097 . Variant in CYP2E1 (rs2031920) was not associated with PD risk. Variants in NAT2  or ABCB1  were not associated with PD risk either.In the secondary analysis, by including 2781 PD cases and 3176 controls in DM and RM, 3.1.4HFE, MTHFR) were included for quantitative analysis. We failed to identify the association between HFE rs1800562, rs1799945, MTHFR rs1801133, rs1801131 and PD risk in all three models .Four variants in two genes  in the pooled analysis. BST1 rs11931532 was negatively related to PD risk in pooled analysis on 1868 PD patients and 3782 controls . In the further analysis, the significant results were also presented in RM about BST1 rs11931532 . By including 3041 PD cases and 4318 controls, we found that HLA-DRB rs660895 was associated with PD risk negatively in all models . CCDC62 rs12817488 was also associated with decreased PD risk in quantitative analysis on 1608 PD cases and 1649 controls in all models .We included four variants in three genes  as exposure and PD as outcome, the primary outcome of IVW model showed that an increased level of ADP-ribosyl cyclase was causally associated with the higher risk of PD  (p =0.07)  as exposure and PD as outcome, the primary outcome of IVW model showed that the higher risk of PD was significantly associated with a decreased level of PON1, which indicated PON1 probably had protective effect on PD risk  (p =0.11)  had causal effect on higher PD risk while PON1(coded by PON1) shown probably protective effect on PD risk. This study may help us to have a deeper understanding of the relationship between the inflammatory variations and PD, and potentially identify biomarkers and create anti-inflammatory and immunological therapy options for PD.The immune system’s dysregulation and inflammatory reactions are now more clearly linked to PD . HoweverIL-1β, TNF-α were not connected with PD risk, variations in IL-1α, IL-6, IL-8, IL-10, and IL-18 were associated  of patients with PD and in the striatum of post-mortem brains from patients with PD , which was inconsistent with the conclusion of previous meta-analysis conducted by Liu Y. et al.  is a signal transduction molecule downstream of the dopamine receptors, which is synthesized from β-NAD+ by both cytosolic and membrane-bound forms of ADP-ribosyl cyclase and/or CD38 , 172. Hil damage . Thus, aOur findings indicated that the onset and progression of PD is closely related to inflammatory response and the disorder of immune system. It would be worth mentioning that PD and atypical parkinsonian syndromes (APS), including multiple system atrophy (MSA) and progressive supranuclear palsy (PSP), have overlapping symptoms that are difficult to make an early diagnose. Nevertheless, PD and APS have different inflammatory patterns, which would be helpful in the differential diagnosis and understanding of the pathogenesis of diseases. Although the occurrence of PSP is also related to the activation of microglia and neuroinflammation, the process may be associated with the accumulation of phosphorylated tau protein, but not α-syn . CompareNrf2 (the transcription factor of NFE2L2) could alleviate the progression of neurodegenerative diseases by counteracting oxidative stress and inflammation (Excitingly, the existence of these inflammatory patterns and immune system alterations provides new insights into anti-inflammatory and immunological treatment strategies for neurodegenerative diseases, including PD. Firstly, inhibiting or activating the function of inflammatory genes might delay or halt disease progression during prodrome or prevent disease progression. For example, activation of ammation . Besidesammation .Currentlammation . Since tammation . Furtherammation , 186. Clammation , 188. SoNevertheless, it must be admitted that our study has several inescapable limitations. Because we combined all of the reported patients and controls for our quantitative analysis, these cases and controls may not be age or sex matched, which might lead to selection bias. Differences in race might also cause confusion. We were unable to run a subgroup analysis on the variables because of the dearth of data. Furthermore, barely fewer than 5 publications were included in some of our quantitative analysis. To reach a reliable conclusion, further unique investigations are required. Due to the insufficient GWAS data resources, we did not conduct causal analysis for all the proteins encoded by statistically significant genes, only ADP-ribosyl cyclase and PON1 were analyzed.IL-1α, IL-1β, IL-10, MnSOD, NFE2L2, CYP2E1, NOS1, NAT2, ABCB1, HFE or MTHFR were not connected to PD risk, we discovered that multiple polymorphisms from IL-6, TNF-α, PON1, CYP2D6, HLA-DRB, BST1, and CCDC62 were statistically correlated with PD risk. Additionally, we indicated the changes in plasm ADP-ribosyl cyclase and PON1 level have causal effects on the risk of PD. Further researches are needed to confirm these findings.In conclusion, we included 18 inflammatory genes, including genes encoding cytokines, genes implicated in oxidative stress, genes for neurotoxins and metabolism-related enzymes, and inflammatory polymorphic loci discovered by GWAS analysis that are strongly connected with PD pathogenesis. While variations in The original contributions presented in the study are included in the article/Study design: MY and YZ. Data collection: MY, JL, SJ, BL, LS, ZH, and YZ. Data analysis: MY, JL, SJ, BL, LS, ZH, and YZ. Writing: MY, JL, SJ, BL, ZH, LS, and YZ. Funding: MY and YZ. Administration: MY and YZ. All authors contributed to the article and approved the submitted version.


The present study results concluded that MUR had no positive effect on the growth performance of the birds. Still, it can improve breast muscles’ fatty acid profile, immune status, and blood biochemistry of broiler chickens.Adding exogenous enzymes in animal diets is a widespread technique with the advantages of lowering antinutritional compounds and increasing nutrient availability and performance. This study evaluated dietary microbial muramidase with four levels , each containing MUR at levels of 0 (control group), 200, 400, and 600 mg Kg−1 diet, with enzyme activity 0, 12,000, 24,000, and 36,000 LSU(F)/kg diet, respectively. The 35-day experiment was completed. The findings showed that adding MUR to broiler meals in amounts of 200, 400, or 600 mg/kg had no impact on growth performance (p > 0.05) during the periods of 4–10, 11–23, and 24–35 days of age. MUR supplementation quadratically impacted the feed conversion ratio of broiler chicks at 11 and 23 days of age (p = 0.02). MUR addition to the diet significantly and level-dependently enhanced the percentage of n−3 and n−6 polyunsaturated fatty acids (PUFA) in breast muscles (p ≤ 0.01), with no alterations to the sensory characteristics of the breast muscles. Dietary MUR increased most of the morphometric dimensions of the small intestine, with the best results recorded at the 200 and 400 mg Kg−1 levels. MUR supplementation at 200, 400, and 600 mg kg−1 linearly lowered the total cholesterol, triglycerides, and low-density lipoprotein cholesterol level (p < 0.01). Still, it significantly increased the high-density lipoprotein cholesterol and very-low-density lipoprotein cholesterol contents compared with the unsupplemented group. Compared to controls, there was a substantial rise in the blood concentration of total protein, albumin, globulin, IL10, complement 3, and lysozyme activity as MUR levels increased (p < 0.01). Moreover, MUR addition significantly increased the immunoexpression of lymphocyte subpopulation biomarkers. We could conclude that MUR can be added to broiler chicken diets up to 600 mg kg −1 to improve broiler chickens’ fatty acid profile in breast muscles, immunity, and blood biochemistry. MUR addition had no positive influence on the bird’s growth.The study aimed to explore how broiler chickens’ blood biochemistry, breast muscles’ fatty acid profile, growth, intestinal morphology, and immune status would be influenced by adding microbial muramidase (MUR) to the diet. Four hundred 3-day-old male broiler chickens were allocated to a completely randomized design consisting of four nutritional treatments ( Poultry farmers have emphasized food safety and bird welfare during the past ten years. Consumer stress pushes for removing synthetic feed additives and reducing unnecessary breeding techniques to protect the environment. Recent poultry production’s two main objectives are high feed efficiency and rapid growth rate. For ideal bird performance, critical aspects influencing gastrointestinal health, such as the bird’s genetic potential, diet quality, environmental circumstances, and disease prevalence, must be considered . Also, tIn animal nutrition, dietary supplementation with exogenous enzymes is a widespread technique. Enzymes are crucial in lowering antinutritional components and increasing the availability of the indigestible portions of the meal for absorption by the animal. Enzyme supplementation can maximize the nutritional content of meals, lowering feed costs, enhancing animal performance throughout growth, and lowering environmental pollution .Peptidoglycans (PGNs) comprise the bacterial cell wall and are considered maintained products of the metabolism and activity of bacteria in the gastrointestinal tract. During normal bacterial cell wall recycling, enormous PGNs are shed from bacterial cell wall fragments in the gut so that PGNs can have a pro-inflammatory effect on the host’s gastrointestinal tract ,7.Escherichia coli, Clostridium perfringens, etc., on the antioxidant status of the gut, involving a rise in intestinal glutathione peroxidase gene expression, and on non-specific immunity in the gastrointestinal tract, involving a rise in gene expression of intestinal interleukin 10 and 18, and interferon-gamma [Lactobacillus spp. [Muramidase  is a glycosyl hydrolytic enzyme that slashes the β-1, 4 glycosidic linkages between N-acetylmuramic acid and N-acetyl glucosamine in the carbohydrate backbone of bacterial PGNs. The primary dissociation products of PGNs are muropeptides that have been informed to alter the inflammatory response in the gastrointestinal tract of various species. But, the extent and form of inflammatory responses modified by mRNA peptides differ according to the cell type . The beson-gamma ,15. Borolus spp. . Microbilus spp. .More attention has been paid in recent years to the expansion of microbial enzymes as potential replacements for antibiotic growth promoters. Several studies have also been performed, with varying degrees of success, on how dietary enzymes impact poultry productivity ,22,23,24Trichoderma reesei (Accession number DSM 32338), Balancius™, Isando. The enzyme activity in the product is 60000 LSU(F)/g. Microbial muramidase’s  safety for chicken feeding was previously established [This research was conducted in a poultry research unit at the faculty of veterinary medicine at Zagazig University, Zagazig, Egypt. Microbial muramidase, EC Number 3.2.1.17, lysozyme or N-acetylmuramidase, is produced by fermentation with a genetically modified strain of ablished .−1 diet, with enzyme activity 0, 12,000, 24,000, and 36,000 LSU(F)/kg diet, respectively. Feed and water were provided ad libitum to the birds. The proximate chemical composition of the basal diet during the starter (4–10 days), grower (11–23 days), and finisher periods (24–35 days) is shown in 2) where the temperature was pointed at 34 °C during the 1 week and regularly lowered till it reached 25 °C at the end of the experiment. Standard vaccination programs were implemented against Newcastle (on days 4 and 14) and Gumboro diseases (on days 7 and 22).Four hundred 1-day-old male Ross 308 broiler chicks were purchased from a commercial hatchery. Before the experiment, birds were subjected to a 3-day adaptation period to achieve a mean body weight (BW) of 92.68 ± 0.17 g. Then the birds were randomly allotted into 4 treatments . The birds were fed baseline diets supplemented with MUR at four levels: 0, 200, 400, and 600 mg KgChicks were weighed at the beginning of the experiment (day 4) to determine the initial body weight (IBW). Then, the body weight and feed intake (FI) were measured at the end of each feeding period to calculate BW gain (BWG), FI, and feed conversion ratio (FCR).The protein efficiency ratio (PER) was calculated according to McDonald et al. .PER=LivThe relative growth rate (RGR) was calculated according to Brody .RGR=FinTen chicks in each group were euthanized using cervical dislocation at the end of this study . A trainA reflectance colorimeter 2 was used to quantify the lightness, redness, and yellowness components of the CIE system color profile while using illumination source C . ThroughUsing a Warner Bratzler (WB) single-blade attachment on a Model 1122 Instron Universal testing equipment, the shear value of the breast muscle was calculated according to Lyon and Lyon .We randomly selected ten birds/group and euthanized them by cervical dislocation at the end of the experiment . Blood wv/v) [We extracted the oils from the breast muscle (5 samples/group) with a solvent mixture of chloroform/methanol  . We measv/v) .n = 10, 2 cm) were preserved in 10% neutral buffered formaldehyde for 72 h, dehydrated, cleared, embedded in wax, sliced with a microtome  into 4 µm cross-sections and longitudinal sections, and stained using hematoxylin and eosin (H&E) [Small intestine samples (in (H&E) . The morin (H&E) We used Bio-spectrum colorimetric diagnostic kits  the determine the serum levels of total cholesterol (TC), triglycerides (TG), and high-density lipoprotein cholesterol (HDL-C). Low-density lipoprotein-cholesterol (LDL-C) was calculated by the Iranian formula, LDL-C= TC/1.19 + TG/1.9–HDL/1.1–38. Using the turbidimetry method , the verWe measured the serum level of complement 3 (C3) using a sandwich enzyme-linked immunosorbent assay . The serum lysozyme activity was determined according to Lie et al. . We measAccording to Grant  and DoumTen spleen samples/group were used to assess the immunoexpression of CD3 and CD20, according to Saber et al. . Slides p < 0.05.The statistical analysis was performed using a completely randomized design and the general linear model (GLM) procedure of SAS 9.2. The linear and quadratic effects of increasing inclusion levels were determined using orthogonal polynomial contrasts. Data variance was expressed as pooled SEM, and the significance level was set at Statistical model:p > 0.05) during the periods from 4–10, 11–23, and 24–35 days of age. MUR quadratically impacted the FCR of broiler chicks at 11 and 23 days of age  among the treatments . The percentages of eicosapentaenoic acid, docosapentaenoic acid, and linoleic acid were increased (Lin. and Quad. p < 0.05) in the breast muscles of birds fed MUR-supplemented diets . The jejunal VH was higher in the MUR400 group (p < 0.01). The duodenal VW, jejunal VW, and ileal VH and VW were increased in all MUR-supplemented groups (p < 0.01). The duodenal CD and MCT were raised in the MUR200 and MUR 600 groups (p < 0.01). The VH:CD ratio was higher in the MUR400 group in the duodenum, the MUR200 and MUR400 groups in the jejunum, and the MUR400 and MUR600 in the ileum (p < 0.01). The jejunal CD and MCT were decreased by MUR supplementation (p < 0.01). The ileal MCT was increased in the MUR600 group and lowered in the MUR200 group (p < 0.01). Moderate goblet cell count in the duodenum was reported in MUR0 and MUR200 groups  and comparatively higher numbers in MUR400 and MUR600 groups .The dimensions of the villous width (VW), villous height (VH), the crypt of Lieberkühn depth (CD), and muscular coat thickness (MCT) were recorded in the duodenum, jejunum, and ileum with the best dimensions were seen in MUR200 and MUR400 groups followed by MUR0 and MUR600 groups . In deta−1 linearly reduced the TC, TG, and LDL-C levels. Still, they significantly increased the HDL-C and VLDL-C compared with the MUR0 group . In addition, linear and quadratic increase in the IL10 and complement 3 levels in the serum of birds fed MUR-supplemented diets  were examined using leukocyte-specific markers (CD3 and CD20). Examined spleen sections showed an average percentage of positive cells per 3 high power fields (HPF) to CD3 (T- cell marker) as follows; 0.93, 1.13 1.86, and 7.22% for MUR0, MUR200, MUR400, MUR600 groups, respectively  function and health are significant factors in attaining animal welfare, feed efficiency, and sustainability . The curBacterial cell walls (PGNs) have rigid structures that are hard to break and are created by continual N-acetylmuramic acid sequences linked with N-acetyl glucosamine by β-1,4 bonds . In a tyThe PGN polymer is degraded during the growth and development by various PGN hydrolases, such as peptidases, muramidase, amidase, and glucosamidase (produced by the host and the associated microbiota). Consequently, muropeptides  are eliminated from the cell wall into the gut . It has Eimeria maxima and C. perfringens, adverse health effects from necrotic enteritis were reduced by feeding on diets containing lysozyme-based antimicrobial blends [The present study showed a substantial and level-dependent rise in the blood concentration of total protein, albumin, globulin, IL10, and complement 3 levels and lysozyme activity in the serum of birds fed diets supplemented with MUR. Moreover, MUR dietary addition significantly increased the immunoexpression of C3 and CD20 in the spleen tissues. Lymphocyte subpopulations are identified by the expression of specific cell surface biomarkers. After antigen recognition, helper and cytotoxic T cells are activated through a signaling cascade initiated by the T-cell co-receptor CD3 . Regulatl blends . MUR wasl blends .−1, with enzyme activity 0, 12,000, 24,000, and 36,000 LSU(F)/kg diet, respectively) in broiler chicken diets. The results concluded that MUR addition resulted in significant intestinal responses in terms of VW, VH, CD, and MCT dimensions in the duodenum, jejunum, and ileum, with the best dimensions seen in the MUR200 and MUR400 groups, without affecting the bird’s growth. MUR-supplemented diets helped enrich the breast meat of broiler chickens with n−3 and n−6 PUFA without altering the sensory characteristics of the carcass, which could increase consumer acceptance. Moreover, Increased supplementation levels of MUR reduced the total cholesterol, triglycerides, and LDL-C while considerably raising HDL and VLDL cholesterol, indicating a hypolipidemic effect. Dietary MUR boosted the birds’ immune systems by improving the blood proteinogram, increasing lysozyme activity, IL10 and C3 concentrations, and upregulating CD3 and CD20 immunoexpression in the spleen tissues.The current study evaluated the dietary addition of MUR (0, 200, 400, 600 mg Kg


Arteriolosclerosis cerebral small vessel disease (CSVD) is a common type of CSVD. This study aimed to explore the factors associated with cognitive function and total MRI burden related to the disease.The demographic characteristics, clinical manifestations, cognitive function score, Barthel Index (BI), blood test index, and follow-up results of arteriolosclerosis CSVD patients treated for the first time in our hospital from January 2014 to August 2022 were collected. White matter hyperintensity (WMH) Fazekas score, total MRI burden, and cerebral atrophy grade were evaluated according to brain MRI findings. Factors associated with CSVD cognitive function were analyzed by binary logistic regression. The correlative factors related to the total MRI burden of CSVD were analyzed by ordered multiple logistic regression.P < 0.001). There were also significant differences in total MRI burden (P = 0.001), WMH Fazekas score, and cerebral atrophy grade (P < 0.001). Mean age (P = 0.012), median deep WMH Fazekas score (P = 0.028), and median deep (P < 0.001) and superficial (P =0.002) cerebral atrophy grade of patients with cognitive impairment at first admission were all higher than those with non-cognitive impairment. Multivariate analysis showed that deep cerebral atrophy was independently and significantly associated with cognitive impairment of CSVD (P = 0.024), and hypertension was significantly and independently associated with total MRI burden (P = 0.001).A total of 146 patients were included in this study, of which 132 cases (90.4%) had hypertension. There were 108 patients (74.0%) with cognitive dysfunction, 97 patients (66.4%) with balance and gait disorders, and 83 patients (56.8%) with moderate-to-severe dependence in daily life (BI ≤ 60 points). Of 146 patients, 79 (54.1%) completed clinical and imaging follow-ups for a median of 3 years. The number of patients with cognitive impairment and BI ≤ 60 points after follow-up significantly increased compared with the first admission (The disease course of arteriolosclerosis CSVD may be related to cognitive function and total MRI burden. Deep cerebral atrophy was an independent risk factor for cognitive dysfunction in arteriolosclerosis CSVD, and hypertension was an independent risk factor for total MRI burden. Cerebral small-vessel disease (CSVD) refers to a series of clinical, imaging, and pathological syndromes caused by various etiologies affecting cerebral arterioles and their distal branches, arterioles, capillaries, capillary vein, and venules. The common etiological types of CSVD include arteriolosclerosis, sporadic or inherited cerebral amyloid angiopathy (CAA), other inherited cerebral small vascular lesions, inflammation, immune-mediated small vascular disease, venous collagen disease, and other small vascular diseases Pantoni, . The twoCSVD is a common cause of dementia and stroke. According to stroke classification, CSVD caused by small artery occlusion accounts for about 30% of the causes of ischemic stroke in China  PVS: fluid-filled spaces that follow the typical course of a vessel as it goes through gray or white matter. The spaces have a signal intensity similar to CSF on all sequences. Because they follow the course of penetrating vessels, they appear linear when imaged parallel to the course of the vessel, and round or ovoid, with a diameter generally smaller than 3 mm, when imaged perpendicular to the course of the vessel. (6) Cerebral atrophy: a lower brain volume that is not related to a specific macroscopic focal injury such as trauma or infarction. Exclusion criteria included the following: (1) symptomatic intracranial or extracranial large artery (diameter > 2 mm) stenosis and occlusions, asymptomatic extracranial carotid artery stenosis ≥ 70%; (2) non-lacunar infarction (infarct lesion with maximum diameter > 20 mm) caused by large vascular lesions  or cardiogenic embolism, cerebral amyloid vascular disease, aneurysms or arteriovenous malformations, and other cerebral bleeds; (3) special causes of white matter lesions ; (4) brain lesions ; (5) severe mental illness, epilepsy, alcohol or drug abuse, intoxication, and metabolic abnormalities; and (6) inflammation- or immune-mediated small cerebral vascular disease, venous collagen disease, or hereditary CSVD.This was a retrospective study. From January 2014 to August 2022, 146 patients with arteriolosclerosis CSVD were first hospitalized in Wujin Hospital affiliated to Jiangsu University, and were included in this study. Inclusion criteria included brain MRI indicating at least one of the following manifestations: (1) RSSI: neuroimaging evidence of recent infarction in the territory of one perforating arteriole, with imaging features or clinical symptoms consistent with a lesion occurring in the previous few weeks, the maximum diameter of lesion < 20 mm in the axial plane. (2) Lacune of vascular origin: a round or ovoid, subcortical, fluid-filled cavity  of between 3 mm and about 15 mm in diameter, consistent with a previous acute small subcortical infarct or hemorrhage in the territory of one perforating arteriole. (3) WMH of vascular origin: signal abnormality of variable size in the white matter that shows hyperintensity on T2-weighted images such as fluid-attenuated inversion recovery, without cavitation . Lesions in the subcortical gray matter or brainstem are not included in this category. (4) CMB: small  areas of the signal void with associated blooming seen on T2All patients had well-documented medical histories, complete laboratory examinations , and brain MRI, CT angiography, or magnetic resonance angiography examinations. In this study, all personal identifiers were removed or anonymized during data collection and analysis to protect the privacy of the participants. The study was conducted under the Declaration of Helsinki, and all participants provided informed consent.General characteristics of all patients were collected based on medical records, including age, sex, education, body mass index (BMI), smoking, hypertension, diabetes, hypercholesterolemia, hyperhomocysteinemia, stroke, transient ischemic attack, ischemic heart disease, and history of atrial fibrillation. Hypertension was defined as blood pressure ≥ 140/90 mmHg, diabetes as fasting blood glucose ≥ 7.0 mmol/L, hypercholesterolemia as total cholesterol ≥ 5.18 mmol/L or LDL cholesterol > 3.12 mmol/L, hyperhomocysteinemia as homocysteine (Hcy) > 15 μmol/L, ischemic heart disease as myocardial infarction, angina, or evidence of myocardial ischemia on the electrocardiogram (ECG), atrial fibrillation as previously diagnosed or found on the ECG, and smoking as currently smoking. At the same time, the main clinical manifestations (core symptoms) of all patients were recorded, and laboratory tests were performed within 7 days after admission, including routine blood tests, liver and kidney function, fasting blood glucose, blood lipid, folic acid, vitamin B12, and homocysteine. Cerebrovascular status was assessed by carotid Doppler ultrasound, CTA, or MRA. Asymptomatic extracranial carotid stenosis < 70% of the patients were still enrolled in the study. The Barthel Index (BI) scale was used to evaluate the activities of daily living (ADL), with a score ranging from 0 to 100 and a higher score indicating greater independence , including axial diffusion-weighted imaging (DWI) scan, with a pulse repetition time (TR), 3,000 milliseconds; echo time (TE), 79.5 milliseconds; thickness, 5 mm; gap, 1.5 mm; and matrix, 210 × 256; axial T1-weighted imaging (T1WI) scan, with TR, 451 milliseconds; TE, 12.5 milliseconds; thickness, 5 mm; gap, 1.5 mm; and matrix, 214 × 352; axial T2-weighted imaging (T2WI) scan, with TR, 4,500 milliseconds; TE, 82.9 milliseconds; thickness, 5 mm; gap, 1.5 mm; and matrix, 233 × 384; axial fluid-attenuated inversion recovery (FLAIR) scan, with TR, 8,000 milliseconds; TE, 75.9 milliseconds; thickness, 5 mm; gap, 1.5 mm; and matrix, 155 × 256; and axial susceptibility weighted imaging (SWI) scan, with TR, 50.1 milliseconds; TE, 40 milliseconds; thickness, 4 mm; gap, 2 mm; and matrix, 194 × 320. All axial scan planes were parallel to the anterior commissure–posterior commissure (AC/PC) line.All MRI images were assessed blinded to clinical information by one experienced neuroradiologist and one neurologist for the presence, location, and size of the recent symptomatic infarct and any other vascular lesions. (1) RSSI: the imaging features were round or ovoid lesions located in the area of perforating arteries, with low signal on the T1WI sequence, with a hyperintense area on DWI, T2WI, and FLAIR. The diameter of the lesions was 3–20 mm. The lesions were mainly distributed in the basal ganglia, internal capsule, centrum semiovale, or brainstem. (2) Lacune: a round or ovoid, in the basal ganglia, internal capsule, centrum semiovale, or brainstem, a fluid-filled cavity of between 3 mm and about 15 mm in diameter, with a CSF signal intensity on T1WI, T2WI, and FLAIR, generally with a hyperintense rim on FLAIR and no increased signal on DWI. (3) WMH: in the early stage, it was a small cap lesion in the frontal lobe and the occipital horn. The progression of the lesion may extend to the subcortical white matter area and fuse, showing abnormal signals of varying sizes in the white matter area of the brain. It offers a high signal on T2WI and FLAIR and an equal or low signal on T1WI. (4) CMB: with the paramagnetic properties of hemosiderin-containing blood breakdown products. Small round or ovoid lesion, well-defined, and homogenous signal loss foci were seen on SWI in the cerebellum, brainstem, basal ganglia, white matter, or cortico-subcortical junction. However, it was not visible on FLAIR, T1WI, and T2WI. The diameter ranges from 2 to 5 mm, with a maximum of 10 mm, differentiated from vessel flow voids and mineral depositions in the globi pallidi. (5) PVS: they appeared round in the axial section and linear if in longitudinal sections and were smaller than 3 mm wide. They were of high signal on T2WI and low signal on T1WI and FLAIR sequences. They were distinguished from lacunes by the latter's large size (more than 3 mm and spheroid shape). PVS mainly occurred in the basal ganglia, subcortical, and brain stem, but rarely in the cerebellum. (6) Cerebral atrophy: it was classified as both deep (enlargement of the ventricles) and superficial (enlargement of the sulci), and MRI showed that the brain volume was reduced, the ventricle was enlarged, and the sulci and gyrus were widened.A score scale proposed by Staals et al.  was usedCerebral atrophy is characterized by reduced brain volume and is not associated with local volume reduction caused by brain trauma and cerebral infarction. Cerebral atrophy was classified as either deep (enlargement of the ventricles) or superficial (enlargement of the sulci). Reduced brain volume, enlarged ventricles, and widened sulci were seen on brain MRI. The sulcal prominence and ventricular size in each individual were assessed on a semiquantitative 10-point scale (grades of 0–9) using predefined visual standards from the Cardiovascular Health Study  and 95% confidence interval (CI). Analyses were performed with SPSS version 21.0, and statistical significance was set at P < 0.05.Continuous variables were described as means ± standard deviations or medians  based on whether data were normally distributed or not. Categorical variables were presented as frequencies and percentages. Two independent sample A total of 146 patients with arteriolosclerosis CSVD were enrolled in this study from January 2014 to August 2022. The mean age of the patients was 75.13 ± 8.93 years old (age range: 46–100 years old), 88 cases (60.3%) were male, 132 cases (90.4%) were complicated with hypertension, 40 patients (27.4%) presented with diabetes mellitus, 27 cases (18.5%) presented with hypercholesterolemia, 71 cases (48.6%) presented with hyperhomocysteinemia, and 57 patients (39.0%) were overweight. There were 17 cases (11.6%) with asymptomatic extracranial carotid artery stenosis on the first admission. There were 123 cases (84.2%) of transient ischemic attack (TIA) or stroke in previous or first diagnosis , includiAt the first admission of all patients, 97 patients (66.4%) had balance and gait disorders, 69 patients (47.3%) had limb weakness, 65 patients (44.5%) had speech disorders, and 25 patients (17.1%) had dizziness or headache. Additionally, 12 patients (8.2%) had emotional and behavioral abnormalities, and 11 patients (7.5%) had dysphagia. According to the ADL assessment, 83 patients (56.8%) had moderate-to-severe dependence in daily life (BI ≤ 60). MMSE assessment indicated that 108 patients (74.0%) had cognitive impairment .A brain MRI analysis of all patients at the first admission revealed that 73 cases (50.0%) had RSSI, six cases (4.1%) had a cerebral hemorrhage, 144 cases (98.6%) had lacune, and 136 cases (93.2%) had CMB. Meanwhile, 73 patients (50.0%) had PVS (grade 2–4), 145 cases (99.3%) had WMH, the total WMH Fazekas score median was 5 (IQR 4–5), and the periventricular WMH Fazekas score median was 3 (IQR 3–3). The median deep WMH Fazekas score was 2 (IQR 1–2). The median score of total MRI burden was 3 (IQR 3–4) points, including 53 cases (36.3%) with four points, 80 patients (54.8%) with three points, and 13 cases with two points (8.9%). Furthermore, 146 cases (100.0%) all had different degrees of cerebral atrophy according to the visual standards from the Cardiovascular Health Study, with a median superficial cerebral atrophy grade of 4 (IQR 3–5) and a median deep cerebral atrophy grade of 5 (IQR 4–6) .P < 0.001). There was also a significant increase in the number of people with moderate-to-severe dependence on daily living requirements compared with the first admission . During follow-up, the WMH range and the degree of cerebral atrophy progressed with the prolongation of the disease course (P = 0.001) and the WMH Fazekas score (P < 0.001) were significantly different. The median of deep and superficial cerebral atrophy grades was significantly higher than that at the first admission (P < 0.001) (There were 79 (54.1%) patients who completed clinical and imaging follow-up for more than 6 months, of which 75 patients had hypertension. The median clinical and imaging follow-up was 3 (IQR 2–5) years, with 48 patients followed up for more than 3 years and 28 patients followed up for more than 5 years. During the follow-up period, 32 cases  had an average of one to three TIA or stroke events, of which one case had TIA, 29 patients had a new small infarction with a median NIHSS score of 4 (IQR 2–4), and two cases had a cerebral hemorrhage. After a median follow-up of 3 years, the average age of patients increased significantly and the number of patients with cognitive impairment was significantly higher than at the time of initial admission  .P = 0.012). The proportion of severely dependent patients (BI ≤ 60) with cognitive impairment was significantly higher than those with non-cognitive impairment (P < 0.001). There were no significant differences in total MRI burden scores between the two groups (P = 0.226), but the median deep WMH Fazekas score (P = 0.028), deep (P < 0.001), and superficial (P = 0.002) cerebral atrophy grades of patients with cognitive impairment at first admission were all higher than those with non-cognitive impairment. There were no significant differences in other clinical characteristics (P = 0.024)  .P = 0.047) and hypertension . However, there was no significant association with sex, age, diabetes, hypercholesterolemia, hyperhomocysteinemia, TIA or stroke history, or cerebral atrophy. In multivariable analysis, after correction for important confounders, including vascular risk factors, age, smoking, and cerebral atrophy, hypertension was significantly and independently associated with total MRI burden in arteriolosclerosis CSVD  and the association for hypertension becoming stronger in multivariable than in univariate testing  for the publication of any potentially identifiable images or data included in this article.L-LM and W-YC were responsible for the conception and design of the study. MH, A-JM, Z-QL, L-LJ, JZ, and Y-FX collected the clinical data, discussed the results, and contributed to the final version of the manuscript. All authors read and approved the final manuscript, contributed toward data analysis, drafting, revising the manuscript, and agreed to be accountable for all aspects of the study.


Ram semen traits including volume, gross motility, concentration, and percent post-thaw motility are routinely assessed prior to use in artificial breeding programs, as they have been shown to influence conception outcomes. Semen quality and associated traits are complex but heritable, and as such, identifying genes that underlie variability in these traits may help develop alternative means to improve conception outcomes and therefore reproductive efficiency in sheep. Therefore, the aim of this study was to identify genomic regions and associated genes that may significantly influence semen traits like volume, gross motility, concentration, and percent post-thaw motility in Merino sheep. Assessment of over 20 years’ worth of semen collection data identified 35 genomic regions to be significantly associated with Merino ram semen volume, gross motility, concentration, and percent post-thaw motility. A total of 290 candidate genes were identified within genomic regions found to be significantly associated with Merino ram semen traits. All Merino ram semen traits were also found to be lowly heritable, affirming results from previous studies. Validation of candidate genes identified in the current study could provide novel insights into molecular mechanisms contributing to variability in semen associated traits.Ovis aries autosomes (OARs), were significantly associated with either of the four semen traits in this study. A total of 89, 95, 33, and 73 candidate genes were identified, via modified Bonferroni, within the QTLs significantly associated with volume, gross motility, concentration, and percent post-thaw motility, respectively. Among the candidate genes identified, SORD, SH2B1, and NT5E have been previously described to significantly influence spermatogenesis, spermatozoal motility, and high percent post-thaw motility, respectively. Several candidate genes identified could potentially influence ram semen traits based on existing evidence in the literature. As such, validation of these putative candidates may offer the potential to develop future strategies to improve sheep reproductive efficiency. Furthermore, Merino ram semen traits are lowly heritable (0.071–0.139), and thus may be improved by selective breeding.Ram semen traits play a significant role in conception outcomes, which in turn may influence reproductive efficiency and the overall productivity and profitability of sheep enterprises. Since hundreds of ewes may be inseminated from a single ejaculate, it is important to evaluate semen quality prior to use in sheep breeding programs. Given that semen traits have been found to be heritable, genetic variation likely contributes to the variability observed in these traits. Identifying such genetic variants could provide novel insights into the molecular mechanisms underlying variability in semen traits. Therefore, this study aimed to identify quantitative trait loci (QTLs) associated with semen traits in Merino sheep. A genome-wide association study (GWAS) was undertaken using 4506 semen collection records from 246 Merino rams collected between January 2002 and May 2021. The R package RepeatABEL was used to perform a GWAS for semen volume, gross motility, concentration, and percent post-thaw motility. A total of 35 QTLs, located on 16  Reproductive efficiency in livestock depends on multiple factors, including insemination, conception, embryonic development, and parturition, and is a key determinant of farm productivity and profitability ,2. ReproRam semen traits are significantly influenced by a variety of factors, including time of year ,11, nutrSLC9C1 and FUT10 were identified as putative candidate genes for gross motility due to their location within identified QTL regions, and because of previous reports that these genes influenced spermatozoal motility, hyperactivation, and fertilisation, respectively. Similarly, a GWAS involving semen volume and number of sperm collected from Holstein-Friesian bulls identified several candidate genes such as GALC, PHF7, and PRKCD, all of which have been previously identified to play a role in spermatogenesis [PLBD1, OXSR1, and EML5 [In the context of semen traits, previous genome-wide association studies (GWAS) have identified several QTLs and candidate genes in a range of livestock species, including dairy sheep , goats , cattle ogenesis . Similarand EML5 , and poland EML5 , spermatand EML5 , and speand EML5 , respectand EML5 , and theand EML5 . TherefoPhenotype data, including semen volume, gross motility, concentration, and percent post-thaw motility, was obtained from a single commercial artificial breeding company (which had two centres over time). The collection and assessment of semen quality, descriptive statistics, and data quality control have been previously described . The datp-value < 1 × 10−6, and if they were located on sex chromosomes. In total, 58,946 SNPs were used for subsequent GWA analysis. The genomic positions of SNPs were taken from the ovine (Ovis aries) reference genome build (Oar_v3.1).Genotype data for 246 rams included in this study was provided by Sheep Genetics . Animals were genotyped using different Ovine SNP chips ranging from 12k (n = 31), 15k (n = 81), 50k (n = 122), and 700k (n = 12). Low-density chips were subsequently imputed to 50k using a reference population as part of the routine pipeline followed by the Animal Genetics and Breeding Unit (AGBU), which was previously described . BrieflyPreliminary analysis was performed to assess the significance of each fixed effect for individual semen traits via linear regression using the R software package, version 4.1.2 . The fixesiduals .p-values for each SNP, as well as the SNP effect of each marker, were determined via Wald statistics within the RepeatABEL software package. Finally, to accurately account for multiple testing levels, the significance threshold was calculated via modified Bonferroni [The R package RepeatABEL v1.1  was usednferroni  as follop-value < 0.0002). These QTL regions were used to search for candidate genes through the National Centre of Biotechnology Information (NCBI) Genome Data Viewer [Ovis aries) genome assembly (Oar_v3.1). Candidate genes were identified in QTL regions that spanned 0.5 Megabase (Mb) around each significant SNP  explain2) of semen phenotypes was calculated as follows:Heritability and variance components were estimated for semen volume, gross motility, concentration, and percent post-thaw motility using the ‘prefitModel’ in the ‘rGLS’ function, including the GRM, via the R package RepeatABEL . The samEstimates of repeatability (r) were calculated as follows: Several SNPs were found to be significantly associated with each semen trait included in this study, i.e., volume, gross motility, concentration, and percent post-thaw motility. Manhattan plots associated with GWA analysis of each of the semen traits are presented in A total of 35 significant SNPs were found across a total of 16 OARs . Of thosOverall, 143, 141, 97, and 138 candidate genes were identified within significant genomic regions associated with semen volume, gross motility, concentration, and percent post-thaw motility after searching the QTL regions, which spanned 0.5 Megabase (Mb) around each significant SNP associated with semen traits via the NCBI Genome Data Viewer using the Oar_v3.1 genome assembly. Several genes located within QTLs were not annotated or did not have known orthologs, and as such, these genes had a ‘LOC’ prefix. A summary of all these genes is presented in PRDM16, ACTRT2, TRNAC-GCA, TTC34, MMEL1, FAM213B, HES5, PANK4, PLCH2, PEX10, RER1, MORN1, SKI, C12H1orf86, PRKCZ, GABRD, CFAP74, TMEM52, CALML6, gross motility; SPERT, SIAH3, ZC3H13, CPB2, LCP1, LRRC63, KIAA0226L, LRCH1, ESD, TRNAG-CCC, and concentration; SIRT5, GFOD1, TBC1D7, PHACTR1, and percent post-thaw motility; no candidate genes identified within QTL.The heritability estimates for semen volume, gross motility, concentration, and percent post-thaw motility are presented in In sheep, semen traits including concentration , gross msolute carrier family 2 member 8 (SLC2A8), actin-related protein T2 (ACTRT2), ribosomal protein L12 (RPL12), and MIS12 kinetochore complex component (MIS12). SLC2A8, also known as Glucose transporter 8 (GLUT8), is highly expressed in the testes and is located within the acrosome of spermatozoal cells as well as in the plasma membrane [SLC2A8 mRNA synthesis was found to coincide with spermatozoal maturation in the mouse testes. In other murine studies, spermatozoal motility was significantly impaired in SLC2A8 null mice due to reduced mitochondrial function [SLC2A8 null mice had irregular changes to the endometrium during embryo implantation [SLC2A8 may be a putative candidate for ram spermatogenesis and motility. Another such gene, ACTRT2, is reported to be localised in the post-acrosomal region of mouse spermatozoa [ACTRT2 may play a key role in the normal morphological development of the acrosome and membrane integrity, as well as promoting spermatozoal motility. Proteomic analysis of ram semen identified ACTRT2 as being significantly expressed in fresh semen compared to cryopreserved semen [ACTRT2 was significantly downregulated in fresh semen collected during the non-breeding season [ACTRT2 may be a key gene modulating semen quality during the breeding and non-breeding seasons. Another gene, RPL12, is known to be significantly downregulated in bulls with low fertility, as assessed via conception rate following artificial insemination [RPL12 has also been found to be upregulated in ram testes [RPL12 may be a positional candidate gene that plays a key role in promoting ram semen maturation and motility. MIS12 has been previously reported to be enriched within networks associated with chromatin condensation following proteomic analysis of turkey testes [MIS12 may play a crucial role in sperm quality and conception success. Furthermore, proteomic analysis of seminal plasma identified MIS12 as being commonly expressed across all fertile males [MIS12 may be a putative candidate gene within ram seminal plasma that may modulate sperm motility and viability. Thus, associated positional candidate genes identified in QTLs significant for semen volume may also have functional roles in overall semen quality.This study identified a total of 35 QTLs located on 16 OARs, specifically nine, seven, eight, and eleven QTLs for semen volume, gross motility, concentration, and percent post-thaw motility, respectively . The ninmembrane . In the function , while oantation . As suchrmatozoa , as wellrmatozoa . Therefoed semen . In a pig season , which ig season . As timeg season ,61 and sg season ,11, ACTRmination . RPL12 hm testes . Given tm testes , RPL12 my testes . Given ty testes ,67, MIS1le males . As semele males , MIS12 mSH2B adaptor protein 1 (SH2B1), mitogen-activated protein kinase 3 (MAPK3), aldolase, fructose-bisphosphate A (ALDOA), 5′-nucleotidase ecto (NT5E), and spermatid associated protein (SPERT). SH2B1 may be a putative candidate influencing spermatozoal motility since SH2B1 was identified within one of the significant QTLs (18.51–18.87 Mb on Sus scrofa chromosome 3) associated with progressive motility in Duroc boars [SH2B1 in obese mice resulted in reduced motility and hyperactivation compared to control mice [MAPK3, is associated with spermatozoal hyperactivation [MAPK3 was also identified as a common differentially expressed gene (DEG) in semen collected from Merino rams when contrasting breed specific differences in the spermatozoal transcriptome [MAPK3 may be a key putative candidate underlying ram spermatozoal motility. Another candidate, ALDOA, was abundantly expressed in spermatozoa collected from rams with high motility compared to rams with low motility [ALDOA was abundantly expressed in rams with higher pregnancy rates following natural mating compared to rams with lower pregnancy rates across three breeding seasons [ALDOA may contribute to spermatozoa motility in rams and thus may be a key putative candidate for spermatozoal motility. Proteomic analysis of ram spermatozoa identified NT5E to be abundantly expressed in ram semen with high motility following cryopreservation  compared to semen with a low cryo-tolerance [NT5E may be a key functional candidate for spermatozoal motility and cryo-tolerance in sheep. Additionally, NT5E was found to be enriched in both seminal plasma and seminal vesicle fluid of bulls [SPERT, also known as chibby family member 2 (CBY2), was significantly expressed in the testes collected from buffalo during puberty compared to testicular tissue from both pre- and post-pubertal buffalo [SPERT was found to be localised to the acrosome, neck, and midpiece of buffalo spermatozoa. As such, SPERT may be a key candidate for spermatogenesis and spermatozoal morphology, given that it is significantly expressed during puberty and is localised to the sperm head and midpiece.Seven QLTs, located on six different OARs, were significantly associated with gross motility. A total of 95 candidate genes were identified within these seven QTL regions, of which several key genes have been previously associated with spermatogenesis and semen quality, including oc boars . Furtherrol mice . Anothertivation , which itivation . Moreovecriptome , as wellcriptome . Hence, motility . Likewis seasons . This suolerance . Therefoof bulls . Finally buffalo . In the peptidyl arginine deiminase 2 (PADI2), cytochrome P450, family 19 (CYP19), and actin like 8 (ACTL8). PADI2 may be a putative candidate gene influencing spermatozoal concentration and motility since PADI2 is abundantly expressed in the epididymis, predominantly in the caput epididymis and vas deferens following RNA sequencing of murine testes [PADI2 may be a putative candidate associated with spermatozoal concentration and a positional candidate modulating spermatozoal maturation and motility. In the same context, PADI2 was found within one of ten Bos taurus autosomes (BTA), explaining the highest portion of genetic variance for both total and progressive motility (BTA 2:1349.81–1359.81) [PADI2 is located within OAR 2:247.37–248.37, and as such, PADI2 may be a significant putative candidate for spermatozoal motility in Merino sheep. Furthermore, PADI2 was identified within a QTL associated with the number of piglets born dead and alive [PADI2 may play a key role in litter size and post-parturition offspring survival in livestock. Another noteworthy candidate gene, CYP19, is associated with spermatozoal concentration and motility in humans, with polymorphisms of CYP19 resulting in reduced sperm concentration within normozoospermic males [CYP19 to be abundantly enriched in spermatozoa, which had higher embryonic cleavage rates compared to spermatozoa, which had lower cleavage rates [CYP19 may be a key candidate for modulating the concentration of spermatozoa in an ejaculate. Expression of ACTL8 has been reported to be isolated to the testes following a study that undertook exome sequencing in males with non-obstructive azoospermia [ACTL8 were not present in the control patients. Hence, ACTL8 may significantly influence spermatogenesis and spermatozoal concentration within an ejaculate. Thus, noteworthy genes identified in QTLs significantly associated with concentration of Merino ram semen, such as PADI2, CYP19, and ACTL8, may be putative candidates for semen traits like concentration and motility in sheep.Thirty-three candidate genes were identified within eight QTLs, located on six OARs, that were found to be associated with semen concentration. Some candidate genes identified in this study have previously been reported in studies related to spermatogenesis and testicular development, including e testes . In sevee testes ,81,82,83e testes . Hence, 1359.81) . In the nd alive ; hence, ic males . The autic males ,88,89 anic males , as wellic males ,92. A page rates . Therefoospermia , a condiospermia . In the MON1 homolog B, secretory trafficking associated (MON1B), RAB3B, member of the RAS oncogene family (RAB3B), ribosomal protein L30 (RPL30), and sorbitol dehydrogenase (SORD). MON1B, located on OAR 14, was identified within genomic regions associated with both cow- and daughter-conception rates following a GWAS in Holstein bulls with known conception rates [MON1B was found to be one of twelve genes associated with embryonic development following a bovine GWAS for cryotolerance and embryonic development in cattle [MON1B may be an important candidate gene that may modulate conception success. RNA Sequencing identified RAB3B as being significantly expressed in good-quality semen compared to low-quality semen following assessment of semen traits in bulls [RAB3B may be a key putative candidate for modulating spermatozoal motility in sheep. Moreover, RAB3-peptides, including RAB3B, are key regulators of the acrosome reaction in sheep [RAB3B may be an important candidate gene for promoting sperm binding to the ova and promoting conception success. In a bull study, RPL30 was abundantly expressed in bulls with high sperm motility compared to bulls with low motility [RPL30 was up- and downregulated in normozoospermic and asthenozoospermic males, respectively, compared to controls [RPL30 may be a key putative candidate gene modulating spermatozoal motility. Another gene, SORD, was identified within the flagella of sperm extracted from the cauda epididymis and had increased expression during the late stages of spermatogenesis in mice [SORD may play a role in modulating ram spermatozoal motility and cryotolerance and, therefore, may influence post-thaw motility in sheep.A total of eleven QTLs on eight different OARs were found to be significantly associated with percent post-thaw motility. A total of 73 candidate genes were identified within these QTL regions. Of these 73 candidates, several have previously been reported to be associated with semen quality and male-side influence on conception outcomes, including on rates . In a sin cattle . Thus, Min bulls . Hence, in sheep . Given tin sheep , RAB3B mmotility . Moreovecontrols . Hence,  in mice . In the  in mice . Hence, p-value < 0.0002). Attempting to use more stringent thresholds, e.g., based on simpleM  [While there are several published GWAS on semen traits in livestock, only a single study has been published in sheep, which involves identifying QTL regions associated with semen volume, concentration, gross motility, and number of spermatozoa using semen records collected from 429 Assaf rams . There i.000001) , resulte.000001) . In the .000001) . The useIn the present study, significant SNPs associated with semen volume, gross motility, concentration, and percent post-thaw motility accounted for 17.08%, 7.59%, 4.69%, and 28.11% of the additive genetic variance . PreviouIt is worth noting that the resource population used in the current study represents a subset of animals that have previously been used to estimate genetic parameters associated with semen traits . The aniSORD, SH2B1, and NT5E, have been previously found to play crucial roles in spermatogenesis, spermatozoal motility, and high motility following cryopreservation; they may be promising candidate genes for Merino ram semen traits in future studies. Several genes like PADI2, RAB3B, and ALDOA, which have been previously associated with promoting spermatozoal maturation, the acrosome reaction, and conception success, were also identified. As such, it would be beneficial to validate such putative candidate genes via molecular analysis to characterise their influence on semen traits and possibly conception outcomes. Overall, results indicate that Merino semen traits are lowly heritable and that several QTLs influencing variability in these traits exist. Further studies are needed to characterise these QTL regions and the genes contained within them.Overall, 35 QTL regions located on 16 OARs were significantly associated with Merino ram semen volume, gross motility, concentration, and percent post-thaw motility. Several genes identified in the present study, including


Tomato is one of the most important fruits worldwide. It is widely consumed due to its sensory and nutritional attributes. However, like many other industrial crops, it is affected by biotic and abiotic stress factors, reducing its metabolic and physiological processes. Tomato plants possess different mechanisms of stress responses in which hormones have a pivotal role. They are responsible for a complex signaling network, where the antioxidant system (enzymatic and non-enzymatic antioxidants) is crucial for avoiding the excessive damage caused by stress factors. In this sense, it seems that hormones such as ethylene, auxins, brassinosteroids, and salicylic, jasmonic, abscisic, and gibberellic acids, play important roles in increasing antioxidant system and reducing oxidative damage caused by different stressors. Although several studies have been conducted on the stress factors, hormones, and primary metabolites of tomato plants, the effect of endogenous and/or exogenous hormones on the secondary metabolism is still poorly studied, which is paramount for tomato growing management and secondary metabolites production. Thus, this review offers an updated overview of both endogenous biosynthesis and exogenous hormone application in the antioxidant system of tomato plants as a response to biotic and abiotic stress factors. Plants are generally subjected to biotic and abiotic stresses during their growth and development . Among b2O2), superoxide anion (O2−), singlet oxygen (1O2), hydroxyl ion (HO−), peroxyl ion (RO2−), alkoxyl ion (RO−), and organic hydroperoxide (ROOH), at basal levels, regulate plant growth and development. However, they are produced at higher levels under unfavorable conditions as a defense mechanism [ROS, such as hydrogen peroxide  is one of the most important vegetable crops worldwide [Tomato . However, PCs also can activate the antioxidant enzyme system of plants. Like in many other plant products, the PCs composition of tomato fruit depends on several conditions, such as plant tissues, variety, growth and development condition, preharvest, harvest, and postharvest management, etc. Hydroxycinnamic acids  are the most abundant PCs reported in tomato plants [PCs are a diverse group of secondary metabolites having in common at least one benzene ring attached to one or more phenolic hydroxyl group substituents. PCs are generally classified into phenolic acids, flavonoids, xanthones, stilbenes, and lignans . They aro plants ,23. TablCarotenoids are bioactive compounds belonging to the isoprenoid family; they provide yellow to red colors to fruits and vegetables . CarotenAmong vitamins, E and C are the most potent antioxidants in fruits and vegetables . VitaminOn the other hand, vitamin C, or ascorbic acid (AA), is a potent water-soluble antioxidant compound of six-carbon lactone. It is a reducing agent; therefore, it can give electrons to any free radical, changing from ascorbic acid to semidehydroascorbic acid or dehydroascorbic acids, which are more stable compounds with a short life as free radicals . Even thGSH (γ-glutamyl-cysteinyl-glycine) is a key secondary metabolite for plant survival due to its role in ROS control . Under n2−) to O2 and H2O2 [SODs are metalloenzymes responsible for catalyzing the dismutation of superoxide radicals , degrading to H2O and O2 [CAT1 and CAT2) are related to stress factors [CATs are AEs presented in practically all living organisms, having vital roles in plant development and as a response to different stresses . Among AO and O2 . CATs arO and O2 . They haO and O2 ,51. Alth factors ,53,54.2O2 generated by the cytosol and chloroplast while maintaining ASA and GSH reserves in different cellular compartments [2O2 by using ASA as an electron donor to H2O and MDHA, transforming the last compound to DHA [APX1, APX2, APX3, APX4, APX5, APX6, and APX7) have been found [APXs are valuable components of the AEs of plants against biotic and abiotic stresses . They arartments . APXs pod to DHA . Like mad to DHA . In tomaen found .2O2 to H2O or alcohols by the oxidation of GSH or thioredoxin [SlPRX25, SlPRX75, SlPRX81, and SlPRX95 were upregulated in tomato plants infected with the tomato leaf curl New Delhi virus.GPXs are tetrameric enzymes containing seleno-cysteine  or cysteine (plant enzymes) in their active site. GPXs catalyze the reduction of Horedoxin . Generaloredoxin . In planoredoxin . Althougoredoxin ,64. In toredoxin . Howeveroredoxin  pointed 2H4) is a phytohormone associated with the ripening of climacteric fruits. At low concentration, it is responsible for tomato fruit’s color, taste, and flavor development [ACS2, ACS4, ACO1, and ACO3), and it is vital for carotenoid gene expression and its accumulation. ET is also involved in plant growth, development, and stress response, and, due to its gaseous nature, can be easily transported in plants without a carrier [Ethylene . The higher ET production was related to the increase in SlERF5, SlJERF1, and SlERF3 gene expression, causing low OD evaluated by MDA production. This study also indicated that tomato plants treated with AVG  had higher OD in leaf and root than untreated plants. Nevertheless, neither OD nor AEs was affected by ET in wild-type Micro-Tom and its Nr mutant when they were submitted to NaCl (100 mM) and CdCl2 (0.5 mM) as stressors [The effect of ET has been evaluated against biotic and abiotic stresses such as NaCl, COependent ,81. Overependent ,83. Howeependent ,85,86. Nependent . Similarependent  indicatetressors . These ctressors .Helocoverpa zea) invasion, reducing PIN2 and PPOF gene expression, but inducing SA biosynthesis via ERF1 and PR1 upregulation. Overproducing ET tomato seeds (Micro-Tom variety) inoculated with mycorrhiza (Glomus clarum) produced higher values of transcription genes, such as CuZnSOD, CAT, and TPX1, related to AEs production [Bacillus megaterium (PGPB) compared to its Nr counterpart, suggesting the intervention of ET in NEACs biosynthesis [Fusarium oxysporum or its toxin (fusaric acid), upregulation of ET key stress responses genes was observed , which may reduce the OD in tomato plants [Regarding biotic stress factors, the information reported in the literature indicates more consistent results. Tian et al.  stated toduction . Similarynthesis . Recent o plants ,94.2-, and H2O2 after 1, 2, and 6 days of storage, respectively [The effect of ET in tomato fruit has also been evaluated during the cold storage (28–45 days at 7–8 °C). Results indicated that despite the ET increase being accompanied by an increment of lyc content, a significant reduction of AA, PCs, and AC was observed ,96. Simiectively . Thus, iectively ,98.2O2, and TBARS by increasing AEs such as SOD, GPX, APX, GR, and CAT [HKT 1;2, NHX, and SOS1 genes, which regulate stress tolerance due to high salinity concentration [SA is a natural phenolic compound (2-hydroxybenzoic acid) essential for signaling plant hormone immunity . SA play and CAT ,108. Thentration ,110. 2O2 application displayed better AEs and OD responses than SA [2O2, and MDA [Alternaria solani [Fusarium oxysporum [Xanthomonas vesicatoria [Potato virus X [Tomato yellow leaf curl virus, Orobanche [Ralstonia solanacearum [Tomato mottle mosaic virus [During the growth of tomato plants at low temperature (10 °C), exogenous SA (0.5 and 1.0 mM) applied in two varieties (Streenb and Floridat) of tomato seedlings at 15 and 30 days after transplanting (one-true leaf stage) significantly increased AEs (PRX and PPO) and AC and reduced the OD indicators, such as MDA and EL, in leaf and root of tomato plants in a dose-dependent manner. It is worthy of interest that the H than SA . This re and MDA . They po and MDA . The ben and MDA . Moreove and MDA ,116. Fora solani ,118, Fusxysporum ,120,121,icatoria , Potato  virus X , Tomato robanche , Ralstonnacearum , Tomato ic virus , and nemic virus .SA  was also applied to alleviate the effect of cold storage (cell membrane damage) on tomato fruit at the green mature stage. Results indicated that OD  was reduced probably due to the P5CS2 upregulation, which is responsible for proline production . MoreovePAL5, C4H, CHS, and FLS, which are related to the phenolic biosynthesis pathways  [Jasmonic acid (JA) and its methyl ester (MeJA) exist naturally in a wide range of higher plants when they are stressed , trigger mainly) ,135,136. mainly) ,137. ACD1. It is important to highlight that, at room temperature, putrescine content did not change between treatments [Moreover, when tomato plants were submitted to cold stress, MeJA application significantly reduced OD (MDA and EL) by increasing putrescine biosynthesis through upregulation of eatments . In thiseatments ,139, whieatments .SIMYC2  [Botrytis cinerea) and treated with exogenous MeJA induces endogenous JA biosynthesis and ET production. The treatment caused an increase in enzymes  related to the synthesis of PCs and POD . Moreover, low relative expression of the SlMYC2 transcription factor was reported, which is interesting because, under some stress conditions such as cold and wound, it is promptly transcribed as a stress response mechanism. However, under pathogen attack, it is a negative regulator of JA response [Jasmonates were also applied to tomato fruit during off-vine ripening to evaluate their effects as postharvest treatment. Tomatoes stored under volatilized MeJA (44.8 µL/L) for 1 week (13 °C) showed an increase in AA, PCs, lyc, and β-carotene after 1 week of retail . Moreoveesponse) . In the response ,143.The plant hormone ABA has been associated with the ripening process of fruits and vegetables ,145. How3 acts as a natural plant growth regulator against biotic and abiotic stress factors by preventing lipid peroxidation and regulating AS [2O2 [GA is a diterpenoid carboxylic acid belonging to the gibberellin family. Among GAs, GAating AS ,159. Resating AS ,161. Intating AS ,161. Regating AS . In the ating AS . On the ating AS ,165. Thi AS [2O2 . 2O2 production. It is interesting to note that IAA effects are dose-dependent because, at low concentrations (<5 µM), an increase in AS is reported, which is caused by an increase in H2O2 production [2O2 biosynthesis [Auxins are phytohormones that play the most relevant role in plant development and growth . Among aoduction ,174,175.ynthesis ,173.p < 0.05) in the AEs  compared to leaves placed at 25 °C [2O2 and MDA) by BR application [2O2, triggering stress tolerance by the upregulation of other hormones, such as ABA.BRs are a group of polyhydroxy steroidal phytohormones present in different parts of plants . BRs parat 25 °C . Later, lication . In thislication  indicateRegarding drought and heavy metal stress (Cd and Cr), BR application improves AS and reduces OD in tomato plants when it was applied in a foliar manner ,182,183.Ralstonia solanacearum) significantly reduced ET biosynthesis and EL and increased AEs [3, IAA, and JA) and reduced the OD caused by cold temperature growth [R. solanacearum [So far, this manuscript has described the effect of individual hormones on AS of stressed tomato plants. However, crosstalk between hormones has also been reported in studies dealing with tomato mutants altered in their hormonal pathways. Information on hormonal crosstalk resulting from responses to stressors is scarce, and valuable information is presented in ased AEs ,124. More growth ,185 and ERFs genes , well-known as stress response factors associated with ET signaling, were upregulated by SA, MeJA, and ABA [LeACS2, LeACS4, LeACC1, LeGR, and LeETR6 genes [BZR1 transcription factor, which directly regulates several genes involved in ET biosynthesis and signaling. Conversely, IAA application significantly reduced ET biosynthesis and, consequently, the ripening process, causing a reduction in lyc and α, β, and δ carotenes by downregulating the PSY1, PSY3, PDS, ZlSO, and CrtiSO genes and chlorophyllase 1-3 [On the other hand, MeJA application significantly increased ET and NEACs biosynthesis in a dose-dependent manner when it was applied against microorganisms in tomato seeds and plants . Interes and ABA ,90,124.  and ABA ,186. In R6 genes  and VardR6 genes  pointed lase 1-3 . Intereslase 1-3 ; howeverPIN2 and PPOF gene expression, two well-characterized wounding and insect response genes in tomato plants [2O2 production in tomato plants, triggering ABA biosynthesis, which increases H2O2 production, improving stress tolerance. Thus, ABA biosynthesis was stimulated by the oxidative stress  in tomato plants, increasing OD and AE by the upregulation of the NCED1 and NCED2 genes [3 and IAA in both resistant and sensitive cold tomato species, while ABA increased in both species. These results confirm the theory about the negative effect of some hormones on each other. However, there exist several interaction nodes among hormones responsible for their up- or downregulation.Ethephon applied to tomato leaves significantly increased (2.6–10.6 times) SA content compared to control, while treatment with MeJA did not affect SA content. Interestingly, when ethephon was applied, MeJA content was reduced by decreasing o plants . On the o plants  pointed D2 genes . They alD2 genes , tomato This review shows that hormones play a pivotal role in the antioxidant response of tomato plants against biotic and abiotic stresses. Tomato plants contain different enzymatic and non-enzymatic antioxidant compounds, which can be regulated by hormones. In general, it seems that, under normal conditions, hormones are found at basal levels; however, under stress conditions, the interaction between ROS and hormones generates a loop, which increases the antioxidant system and alleviates oxidative damage. Moreover, as has been described, some hormones presented a positive, negative, or null effect among them, showing their impacts on molecular and genetic signaling. This review is valuable to clarify some important questions about hormones and their effects on oxidative damage in tomato plants. Nevertheless, further studies are needed to clarify the hormone effects on improving antioxidant responses against stress factors and how to take advantage of promoting resistance or increasing health-promoting compounds found in tomato plants.


Cis-Mendelian randomization (MR) was used to identify plasma proteins associating with CMR traits as well as with any of the following cardiac outcomes: HF, non-ischemic cardiomyopathy, dilated cardiomyopathy (DCM), atrial fibrillation, or coronary heart disease. In total, 33 plasma proteins were prioritized, including repurposing candidates for DCM and/or HF: IL18R (providing indirect evidence for IL18), I17RA, GPC5, LAMC2, PA2GA, CD33, and SLAF7. In addition, 13 of the 25 druggable proteins  could be mapped to compounds with known oncological indications or side effects. These findings provide leads to facilitate drug development for cardiac disease and suggest that cardiotoxicities of several cancer treatments might represent mechanism-based adverse effects.Dysfunction of either the right or left ventricle can lead to heart failure (HF) and subsequent morbidity and mortality. We performed a genome-wide association study (GWAS) of 16 cardiac magnetic resonance (CMR) imaging measurements of biventricular function and structure.  Combining deep-learning of CMR, GWAS, and Mendelian randomization identified 33 plasma proteins affecting cardiac outcomes. Dysfunction of the right or left ventricle , arising due to intrinsic heart muscle disease, coronary artery disease, or pulmonary or systemic hypertension, leads to the clinical syndrome of heart failure (HF)  imaging is the gold standard for quantification of biventricular function and morphology, and has become an integral diagnostic modality for cardiac disease (see table S1). Here, we used CMR images from the UK Biobank (UKB) to extract measures from both LV and RV using a purpose-built deep-learning algorithm  on 16 CMR traits measured in up to 36,548 UKB subjects. Subsequently, we format-normalized protein quantitative trait loci (pQTLs) data sourced from three independent GWAS involving cross-platform measurement of plasma protein concentrations using Somalogic .CMR measurements were obtained from a sample of up to 36,548 UKB subjects using an extensively validated deep-learning approach  years old, and 18,879 (51.8%) were women. Participants had a mean systolic blood pressure (SBP) of 138.2 mmHg , a mean diastolic blood pressure (DBP) of 78.6 mmHg , and a mean heart rate of 62.5 beats per minute (bpm)  (see table S3).−8, and 32 variants passing a more conservative threshold of 7.14 × 10−9, accounting for the number of PCs necessary to explain 90% of the CMR trait variance (figs. S3 and S4). This included lead variants in or around genes known to affect cardiac outcomes, such as BAG3, TTN, SMARCB1, SYNPO2L, TBX5, and IGF1R. Please see Supplementary Results, figs. S4 to S8, and tables S5 to S8 for a full description of the GWAS findings.We performed a GWAS on 16 CMR traits, leveraging genotyped and imputed variants from the Affymetrix BiLEVE and Axiom arrays, and applying BOLT-LMM conditional on age, sex, body surface area (BSA), SBP, genotype measurement batch, 40 principal components (PCs), and assessment center. This resulted in 91 unique lead variants  drugged or druggable: IL6RA, IL8, CO6A1, LYAM1, PA2GA, ISK2, CD33, PPAC, COFA1, GPC5, TIE2, IL18R, LAMC2, I17RA, SLAF7 . From a Twenty-five proteins associated with HF, DCM, or non-ischemic CM . We did observe a significant difference in tissue-specific expression (P = 9.01 × 10−3), with prioritized plasma proteins more frequently highly expressed in spleen, lymph node, liver, granulocytes, kidney, pancreas, and lung tissues (fig. S16). In addition to the cardiac outcomes these proteins were prioritized for, the cis-MR phenome-wide scan showed that these proteins were frequently associated with DBP, SBP, electrocardiography (ECG) measurement during exercise, lipid fractions , estimated glomerular filtration rate (eGFR), body mass index (BMI), glycosylated hemoglobin (HbA1c), C-reactive protein, lung function , and carotid intima-media thickness (cIMT)  of the 25 drugged or druggable proteins were targeted by compounds with a cardiovascular indication or side effect .cis-MR analysis leveraged three distinct plasma pQTL resources, distilling a prioritized set of 33 plasma proteins that affect both CMR traits and cardiac outcomes (P = 0.85). This likely reflects imperfect understanding of the relation between CMR traits and disease. Furthermore, given the strong  correlation among CMR traits, inference might be further improved by considering CMR traits jointly.The outcomes . While tOur analyses have highlighted drug targets that affect multiple cardiac outcomes . Some ofcis-MR of plasma pQTLs, we identified seven proteins that affected PFR as well as HF or DCM risk: UD16, MANBA, TNR5, TNF12, MFGM, CPC5, and BAG3, where the last five proteins were (indirectly) drugged or druggable, providing leads for drug development  of the 25 drugged or druggable proteins were targeted by a compound with an oncological indication . For exacis-MR of protein effect on these CMR measurements, as well as 56 clinically relevant traits. By leveraging orthogonal lines of evidence on mRNA expression, protein interactions, and drug compound indications and side effects, we were able to identify a robust set of proteins affecting CMR and cardiac outcomes. Genetic analyses were conducted using methods such as BOLT-LMM and BOLT-REML, which account for potential population admixture or relatedness  suggesting that most of the findings are true positive.In the current study, we combined GWAS on UKB-derived CMR measurements of RV and LV structure and functions, with cross-platform dataof the plasma proteome, and perform The following potential limitations deserve consideration. We did not exclude individuals with non-European ancestry, and instead, we accounted for potential population stratification bias through linear mixed models  known to affect the LV or RV traits (see table S2).QC) to extract LV and RV CMR measurements has been previously described and extensively validated (The deep-learning methodology (AI-CMRP < 1 × 10−6).We used genotyped and imputed data as provided by UKB (GRCh37 assembly)  was used to estimate narrow-sense genetic heritability , as well as the pairwise genetic correlation between the CMR traits.BOLT-REML . Locus-view plots combined variant-specific CMR associations around each respective lead variant [±250-kbp (kilo–base pair) flanking region] with information on regional genes and their exon structure. These plots were enhanced with an incidence  matrix annotating genes on 23 criteria, including whether the gene was coding, encoded a target for drug compounds with known cardiometabolic (side) effects, and had a cis-MR association between any of the considered plasma proteins and a CMR trait, previous associations with cardiometabolic traits sourced from GWAS Catalog .Lead variants were identified through linkage disequilibrium (LD) clumping within a 1-Mb flanking region, applying a pairwise Genetic associations with plasma protein concentration were available from the following sources: Somalogic measurements on 3301 participants of the INTERVAL cohort , standardizing file structures, mapping variants against the same genome assembly, assigning UniProt identifiers, and providing annotations with Variant Effect Predictor (VEP), Polymorphism Phenotyping v2 (PolyPhen), and Combined Annotation Dependent Depletion (CADD).cis gene known to encode the protein, clumping variants to an LD R-squared of 0.40, where residual LD was modeled using a generalized least square (GLS) model  estimator, as well as with an Egger correction protecting against horizontal pleiotropy , we identified proteins with a CMR association, prioritizing results on druggability and on directional concordance. Druggability was determined through linkage with ChEMBL and BNF. The BNF draws information from drug medication inserts, scientific literature, regulatory authorities, and professional bodies and is jointly authored by the British Medical Association and the Royal Pharmaceutical Society. ChEMBL . Next, we identified the distance between these concordant proteins and the nearest drugged or druggable protein(s) based on the IntAct .LD score regression  with the uniform distribution expected under the null hypothesis  refer to a unit change of the independent variable, typically one SD in plasma protein level (MR results) or an increase in risk allele (GWAS results). To better illustrate concordance, and only where specified, MR results were orientated toward the cardiac beneficial effect direction by multiplying EDV, ESV, EDM, and MVR MR estimates by −1.


The signs and symptoms associated with GAGs varied and include headache (19%), sensory change(s) (11%), and intracranial hypertension (2%), among diverse and potentially serious sequelae. Notably, brain herniation was present within 38 GAGs (22%). Among treated individuals, subsets were managed medically , surgically , and/or by endovascular DVS stenting . Histologic workup of 53 (27%) GAG cases depicted internal inflammation (3%), cystic change consistent with fluid accumulation (2%), venous thrombosis (1%), hemorrhage (1%), meningothelial hyperplasia (1%), lymphatic vascular proliferation (1%), and lymphatic vessel obliteration (1%). This review emphasizes heterogeneity in GAG subtypes, morphology, composite, location, symptomatology, and imaging presentations. Additional systematic investigations are needed to better elucidate the pathobiology, clinical effects, and optimal diagnostic and management strategies for enlarged and symptomatic arachnoid granulation subtypes, as different strategies and size thresholds are likely applicable for medical, interventional, and/or surgical treatment of these structures in distinct brain locations.Giant arachnoid granulations (GAGs) are minimally investigated. Here, we systematically review the available data in published reports to better understand their etiologies, nomenclature, and clinical significance. In the literature, 195 GAGs have been documented in 169 persons of varied ages . Prior reports depict intrasinus  (84%), extrasinus  (15%), and mixed (1%) GAG types that exhibit pedunculated, sessile, or vermiform morphologies. GAG size ranged from 0.4 to 6 cm in maximum dimension  and encompassed symptomatic or non-symptomatic enlarged arachnoid granulations (≥1 cm) as well as symptomatic subcentimeter arachnoid granulations. A significant difference was identified in mean GAG size between sex (females, 1.78 cm; males, 3.39 cm;  Arachnoid granulations (AGs) are macroscopically visible arachnoid tissue outpouchings that protrude into bone, dura, and/or dural venous sinuses (DVSs) . HistoriTypically, AGs are asymptomatic structures that abut dural tissues and measure only a few millimeters in diameter ,6, but tWith the understanding that GAGs represent enlarged AGs, published cases of GAGs were searched using PubMed and Scopus to identify original articles and reviews reported in the English language, up to and including July 2023, using a Boolean search strategy incorporating the following terms: ‘Giant’ OR ‘Large’ AND ‘Arachnoid granulation’ OR ‘Pacchionian granulation’ OR ‘Pacchionian body’. Given the paucity of reports meeting this search strategy, the approach was supplemented by a reverse bibliographic search of published cases, series, and reviews. The literature was reviewed by both investigators, guided by the standards of the Preferred Reporting Items for Systematic Review and Meta-Analysis (PRISMA) . AvailabThe published literature from 1973 to 2023 yielded 41 publications describing GAGs in 169 persons ,44,45,46Of the 147 persons with documented gender, GAGs involved 80 (54%) females and 67 (46%) males and were similarly distributed across sex  . The affp < 0.05) [The size distribution of GAGs is summarized in  < 0.05) ,31,32,33The majority of GAGs presented along SSS or transverse sinuses, or in parasagittal brain regions, whereas a subset involved the temporal bone C and cauFourteen calvarial-type or mixed-type GAGs were evaluated histologically ,15,16,45At least 27 DVS-type GAGs were identified on postmortem DVS examination ,10,15 T. Howeverp < 0.05).While some reports define GAGs as normal AG variants of no known clinical significance ,34, heteSome afflicted persons indicated that symptoms had been ongoing for several years, or for decades prior to diagnosis ,29,45. ADiagnosis of GAG on imaging workup  was accop = 0.04), although the reason for this sex difference is unclear [While the internal GAG characteristics generally paralleled those of CSF on CT and MRI, GAGs more commonly demonstrated internal vascular  and/or soft tissue elements that were not easily observable in smaller AGs. In an imaging review, brain parenchymal herniation into GAGs was found in 22% of DVS-type GAGs . The int unclear .Several individuals experienced misdiagnosis or delayed diagnosis of GAG ,22,43,45Endovascular stenting was performed in seven patients with DVS-type GAGs who failed medical treatment and were found to have DVS stenosis ,29,41,42In total, 15 patients aged 4 months to 73 years underwent craniotomy with resection of DVS-type, calvarial-type, or mixed-type GAGs ,28,32,45At surgery, GAGs appeared as nodular projections or cysts that protruded through the dura, eroded into adjacent calvarium, and caused subcutaneous masses ,45. DuriA literature review highlights the lack of consensus for a precise definition of the term GAG ,22,23,35It has been suggested that AGs and GAGs drain exclusively to the DVS and that their size is an indicator of CSF pressure . But a rWhile some reports define GAGs as incidental AG variants ,34, manyWhile evidence pertaining to GAG anatomy and biology is limited, published data illustrate that these structures are variably symptomatic, may reach extraordinary dimensions , exhibi