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Abstract: Neuropsychiatric disorders that affect the central nervous system cause considerable pressures on the health care system and have a substantial economic burden on modern societies. The present treatments based on available drugs are mostly ineffective and often costly. The molecular process of neuropsychiatric disorders is closely connected to modifying the genetic structures inherited or caused by damage, toxic chemicals, and some current diseases. Gene therapy is presently an experimental concept for neurological disorders. Clinical applications endeavor to alleviate the symptoms, reduce disease progression, and repair defective genes. Implementing gene therapy in inherited and acquired neurological illnesses entails the integration of several scientific disciplines, including virology, neurology, neurosurgery, molecular genetics, and immunology. Genetic manipulation has the power to minimize or cure illness by inducing genetic alterations at endogenous loci. Gene therapy that involves treating the disease by deleting, silencing, or editing defective genes and delivering genetic material to produce therapeutic molecules has excellent potential as a novel approach for treating neuropsychiatric disorders. With the recent advances in gene selection and vector design quality in targeted treatments, gene therapy could be an effective approach. This review article will investigate and report the newest and the most critical molecules and factors in neuropsychiatric disorder gene therapy. Different genome editing techniques available will be evaluated, and the review will highlight preclinical research of genome editing for neuropsychiatric disorders while also evaluating current limitations and potential strategies to overcome genome editing advancements.

The preventive effect of vitamin D against breast cancer can be influenced by gene polymorphisms. This study aimed to investigate the association between serum level of 25(OH) vitamin D and FTO genotype in breast cancer patients. A cross‐sectional study was carried out on 180 newly diagnosed patients with breast cancer in Tehran, Iran. The blood samples were collected from the participants in order to assess the FTO gene rs9939609 polymorphism by the tetra‐primer amplification refractory mutation system (Tetra‐ARMS) PCR method. The serum level of 25(OH) vitamin D was measured using the direct competitive enzyme‐linked immunosorbent assay (ELISA) method. The association between vitamin D and the FTO genotype in patients with breast cancer was assessed after adjustment for cofounders. The frequency of TT, AT and AA genotypes in the breast cancer patients were 43% (n = 77), 49% (n = 89) and 8% (n = 14), respectively. All patients with higher than 40 ng/dl of serum 25(OH) vitamin D had one or two copies of FTO rs9939609 risk allele (p = 0.019). No linear association was found between the number of FTO risk allele and the level of serum vitamin D. All patients with high serum level of 25(OH) vitamin D had one or two copies of FTO rs9939609 risk allele. FTO gene polymorphisms may counteract the beneficial effects of vitamin D in breast cancer prevention. Further studies can help to better understand the genetic factors predisposing to breast cancer and their effect on the association between vitamin D and breast cancer.

Background and AimThe association between the rs9939609 polymorphism of fat mass and obesity-associated gene (FTO) and risk of colorectal cancer is controversial. This study aims to evaluate the relationship between FTO rs9939609 polymorphism and colorectal cancer (CRC) in Iranian people.MethodsA case-control study was conducted on 125 patients with CRC and 250 healthy subjects in Tehran, Iran. Demographic data and blood samples were collected from all participants. Genotyping of rs9939609 polymorphism was performed by the tetra-primer amplification refractory mutation system-polymerase chain reaction (T-ARMS-PCR) method.ResultsThe occurrence of AA genotype of FTO rs9939609 polymorphism in the colorectal cancer patients was significantly higher compared to that of healthy subjects (16.4 vs. 2.9%, respectively, P=0.02). The association between the frequency of risk allele of the FTO polymorphism and CRC (B=1.67, P=0.042) remained significant after adjustment for age. Further adjustment for gender (model 2) and marital status (model 3) did not change this result (B=1.67, P= 0.042 and B=1.67, P=0.043, respectively). The results remained significant after additional adjustment for ethnicity (B=1.57, P= 0.047).ConclusionWe found a positive association between the A allele of the rs9939609 polymorphism and CRC. Future studies are required to identify the underlying mechanisms.

Diabetes epidemiological quantities are demonstrating one of the most important communities' health worries. The essential diabetic difficulties are including cardiomyopathy, nephropathy, inflammation, and retinopathy. Despite developments in glucose decreasing treatments and drugs, these diabetic complications are still ineffectively reversed or prohibited. Several signaling and molecular pathways are vital targets in the new therapies of diabetes. This review assesses the newest researches about the key molecules and signaling pathways as targets of molecular pharmacology in diabetes and diseases related to it for better treatment based on molecular sciences. The disease is not cured by current pharmacological strategies for type 2 diabetes. While several drug combinations are accessible that can efficiently modulate glycemia and mitigate long-term complications, these agents do not reverse pathogenesis, and in practice, they are not established to modify the patient's specific molecular profiling. Therapeutic companies have benefited from human genetics. Genome exploration, which is agnostic to the information that exists, has revealed tens of loci that impact glycemic modulation. The physiological report has begun to examine subtypes of diseases, illustrate heterogeneity and propose biochemical therapeutic pathways.

The purpose of this review is to highlight recent scientific developments and provide an overview of virus self-assembly and viral particle dynamics. Viruses are organized supramolecular structures with distinct yet related features and functions. Plant viruses are extensively used in biotechnology, and virus-like particulate matter is generated by genetic modification. Both provide a material-based means for selective distribution and delivery of drug molecules. Through surface engineering of their capsids, virus-derived nanomaterials facilitate various potential applications for selective drug delivery. Viruses have significant implications in chemotherapy, gene transfer, vaccine production, immunotherapy and molecular imaging., Lay abstract The purpose of this review is to highlight recent scientific developments and provide an overview of virus self-assembly and viral particle dynamics. Viruses are organized supramolecular structures with distinct yet related features and functions. Plant viruses are extensively used in biotechnology, and virus-like particulate matter is generated by genetic modification. Both provide a material-based means for selective distribution and delivery of drug molecules. Through surface engineering of their capsids, virus-derived nanomaterials facilitate various potential applications for selective drug delivery. Viruses have significant implications in chemotherapy, gene transfer, vaccine production, immunotherapy and molecular imaging. Here we performed a comprehensive database search to review findings in this area, demonstrating that viral nanostructures possess unique properties that make them ideal for applications in diagnostics, cell labeling, contrasting agents and drug delivery structures.

Background: Periodontal disease(s) and metabolic illnesses negatively impact the quality of life and, eventually mental health. Objective: This study investigated the effect of Porphyromonas gingivalis (W83) oral infection on the development of Alzheimer’s disease (AD) pathophysiology in a wild-type obese, diabetic (db/db) mouse model. Methods: The db/db mice were either orally infected with P. gingivalis and Fusobacterium nucleatum or sham infected for 16 weeks. The presence of amyloid-β (Aβ) and neurofibrillary tangles (NFTs) were assessed using a silver impregnation technique and subsequently by immunohistochemistry for tau and neuroinflammation. The mRNA abundance of a panel of 184 genes was performed using quantitative real-time PCR, and the differentially expressed genes were analyzed by Ingenuity Pathway Analysis. Results: While no Aβ plaques and NFTs were evident by silver impregnation, immunohistochemistry (glial cell markers) of the P. gingivalis-infected mice tissue sections exhibited neuroinflammation in the form of reactive microglia and astrocytes. Anti-tau immunopositivity, in addition to cells, was prominent in thickened axons of hippocampal CA neurons. The mRNA abundance of crucial genes in the insulin signaling pathway (INSR, IGF1, IRS, IDE, PIK3R, SGK1, GYS, GSK3B, AKT1) were upregulated, potentially exacerbating insulin resistance in the brain by P. gingivalis oral infection. Increased mRNA abundance of several kinases, membrane receptors, transcription factors, and pro-inflammatory mediators indicated hyperactivation of intracellular cascades with potential for tau phosphorylation and Aβ release in the same infection group. Conclusion: P. gingivalis W83 infection of db/db mice provides a disease co-morbidity model with the potential to reproduce AD pathophysiology with induced periodontal disease.

The mortality and morbidity rates for prostate cancer have recently increased to alarming levels, rising higher than lung cancer. Due to a lack of drug targets and molecular probes, existing theranostic techniques are limited. Human LIN28A and its paralog LIN28B overexpression are associated with a number of tumors resulting in a remarkable increase in cancer aggression and poor prognoses. The current review aims to highlight recent work identifying the key roles of LIN28A and LIN28B in prostate cancer, and to instigate further preclinical and clinical research in this important area.

Porphyromonas gingivalis triggers a range of innate immune responses in the host that may contribute to the development of periodontitis and dementing diseases including Alzheimer’s disease (AD). This study aimed to assess the mode of action of trans-resveratrol in modulating the P. gingivalis lipopolysaccharide (PgLPS) induced metabolic inflammation in a neuronal cell model. Confluent IMR-32 neuroblastoma cells were treated with trans-resveratrol from Polygonum cuspidatum in the presence or absence of PgLPS. The abundance of messenger ribo-nucleic acid (mRNA) transcripts of a panel of 92 genes was quantitatively assessed through targeted transcriptome profiling technique and the biochemical pathways affected were identified through Ingenuity Pathway Analysis. Gene expression analysis revealed that trans-resveratrol down-regulated the mRNA of multiple gene markers including growth factors, transcription factors, kinases, trans-membrane receptors, cytokines and enzymes that were otherwise activated by PgLPS treatment of IMR-32 neuroblastoma cells. Pathway analysis demonstrated that the cellular oxidative stress caused by the activation of phosphoinositide-3-kinase/Akt1 (PI3K/Akt1) pathway that leads to the production of reactive oxygen species (ROS), chronic inflammatory response induced by the activation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) pathway and nutrient utilization pathways were favourably modulated by trans-resveratrol in the PgLPS challenged IMR-32 cells. This study demonstrates the potential of trans-resveratrol as a bioactive compound with multiple modes of intracellular action further supporting its therapeutic application in neuroinflammatory diseases.

Background: Potassium bromate (KBrO3), a food additive, has been used in many bakery products as an oxidizing agent. It has been shown to induce renal cancer in many in-vitro and in-vivo experimental models. Objectives: This study evaluated the carcinogenic potential of potassium bromate (KBrO3) and the chemopreventive mechanisms of the anti-oxidant and anti-inflammatory phytochemical, curcumin against KBrO3-induced carcinogenicity. Method: Lactate dehydrogenase (LDH) cytotoxicity assay and morphological characteristics were used to assess curcumin's cytoprotective potential against KBrO3 toxicity. To assess the chemopreventive potential of curcumin against KBrO3-induced oxidative insult, intracellular H2O2 and the nuclear concentration of the DNA adduct 8- OHdG were measured. PCR array, qRT-PCR, and western blot analysis were used to identify dysregulated genes by KBrO3 exposure. Furthermore, immunofluorescence was used to evaluate the ciliary loss and the disturbance of cellular tight junction induced by KBrO3. Results: Oxidative stress assays showed that KBrO3 increased the levels of intracellular H2O2 and the DNA adduct 8-OHdG. Combination of curcumin with KBrO3 efficiently reduced the level of H2O2 and 8-OHdG while upregulating the expression of catalase. PCR array, qRT-PCR, and western blot analysis revealed that KBrO3 dysregulated multiple genes involved in inflammation, proliferation, and apoptosis, namely CTGF, IL-1, and TRAF3. Moreover, qRT-PCR and immunofluorescence studies showed that KBrO3 negatively affected the tight junctional protein (ZO-1) and induced a degeneration of primary ciliary proteins. The negative impact of KBrO3 on cilia was markedly repressed by curcumin. Conclusion: Curcumin could potentially be used as a protective agent against carcinogenicity of KBrO3.

This experiment was conducted to evaluate the effect of feeding regimens for the breeder broiler hen on the performance of broiler chickens. Breeder broiler hens (196 hens and 28 roosters; age between 31 and 38 wk) were randomly assigned to 7 dietary treatments that differ in the proportion of daily allowance served at different times with 1 (100% at 0400 h), 2 (50% at 0400 and 1600, or 75% and 25% at 0400 and 1600, respectively), 3 (33% at 0400, 1200 and 1600, or 50% at 0400 and 25% at 1200 and 1600), and 4 (33% at 0400, 22% at 0800, 1200 and 1600, or 55% at 0400 and 17% at 0800, 1200 and 1600) meals in every 24-h cycle. There were 4 replicates per treatment, each consisted of 7 hens and 1 rooster received experimental diets for 8 wk continuously. The growth performance, feed intake, feed conversion ratio, protein intake, protein conversion ratio, energy intake, and energy conversion ratio in the broiler chicken were evaluated for 42 d. Dietary regimens (frequency and timetable) for breeder broiler hens affected (P < 0.001) the feed intake, feed conversion ratio, protein intake, protein conversion ratio, energy intake, and energy conversion ratio in the offspring (d 0–42). Overall feed conversion, protein conversion and energy conversion in broiler chickens from breeder hens received daily allowances in 3 or 4 meals were improved compared to those from breeder hens received 100% dietary allowance as a single meal. It is concluded that providing the daily diet allowance of breeder broiler hens in 3 or 4 meals is likely to improve growth performance in their offspring.

Sumac (Rhus coriaria L.) is a plant species belong to Anacardiaceous family that is worldwide diffused. The sumac seed power (SSP), produced by grinding dried fruits, is recognized to have defensive and beneficial effects on numerous health‑related problems. In this study, SSP was included in broilers basal‑diet to investigate the comparative effects of different levels of SSP on performance, carcass characteristics, blood parameters, immune system and ileal microorganisms. A total of 225, one day‑old male broilers (Ross 308) were randomly assigned to the five dietary treatments with three replicates per treatment. The experimental diets were: basal‑diet (BD); and BD including 0.05, 0.10, 0.15 and 0.20% SSP, respectively. During the whole feeding period (42 days), birds fed corn‑based grower (1‑21 days) and finisher (22‑42 days) diets, respectively. Results indicated that supplementing SSP had no effect on broiler body weight gain, feed intake and feed conversion as well as carcass characteristics (P0.05). Similarly, blood total protein, albumin, glucose and triglyceride were not influenced by dietary SSP. Conversely, serum total cholesterol and LDL‑cholesterol levels were decreased, while HDL‑cholesterol increased in all SSP fed groups compared to control (P0.05). In this study the addition of SSP in broilers diets did not show any effect on blood heterophils and lymphocyte. Moreover, the lactobacillus count remained unaffected by dietary treatments, while E. coli count in broiler ileal content was lower when fed 0.10% SSP than the other groups (P0.05). Thus, the present findings indicated a positive effect of feeding SSP (especially at 0.10% diet) on blood cholesterol levels and E. coli count in broiler chickens

Grass-based milk production is a major contributor to Irish agricultural output. The study characterized the Irish milk pool using stable isotope ratio analysis of carbon, nitrogen, oxygen and sulphur. Authentic raw milk samples were collected from 50 farms on five occasions over 13 months. Mean values of −27.11, 6.79, −3.27 and 6.16‰ were obtained for δ13C, δ15N, δ18O and δ34S, respectively. δ13C values reflected a high level of grass input and values increased with increasing cereal concentrate feed input (P

The objectives of this experiment were to compare the in-vitro anti-inflammatory activity of crude extracts (cold water (CWE), hot water and 80% ethanol extract) of F. vesiculosus and to predict the key molecular targets of the extract with most anti-inflammatory activity in-vitro . In TNF-α challenged Caco-2 cells, of the three crude extracts, the CWE exhibited maximum inhibition of IL-8 production. In the ex-vivo challenged porcine colonic tissue, CWE inhibited the expression (> 2 fold) of inflammatory mediators ( PTGS2 , C5 , LYZ ), cytokines ( IL17A , IL8 ), chemokines ( CCL2 , CXCL2 , CXCL10 , CXCL11 ), cell adhesion molecules ( ICAM1 , VCAM1 ), toll like receptors ( TLR4 , TLR7 ) and components of NF-κB ( NFKB1 , RELB ), MAPK ( MAP3K8 ) and AP-1 ( CJUN ) pathways. The gene expression analysis suggest that the CWE does contain immunomodulatory bioactive compound/s that mediated through the interferon regulatory system 2, TNF-α inducing protein 3 and TNF-α receptor 2 and thus has potential application for human and animal health.

The anti-inflammatory bioactivities of three crude extracts of Ascophyllum nodosum (cold water, hot water and 80% ethanol extract (EE)) were initially evaluated. Following from this, the immune modulatory efficacy of EE and its MW fractions ( 100 kDa) were compared. In TNF-α challenged Caco-2 cells, the crude extracts reduced IL-8 production ( P 2 fold down-regulation of; cytokines ( IL8 , TNFA , IL1B, IL18 and CSF1 ) chemokines ( CXCL10 and CCL5 ), components of NF-κB pathway ( NFKB2 and IKBKB ), and inflammatory mediators ( PTGS2 and MIF ) genes. In the porcine colonic tissue ex-vivo challenged with lipopolysaccharide, EE and its MW fractions down-regulated the immune related targets LYZ , IL8 , PTGS2 , TLR 6, CXCL10 , IL6 , CXCL11 , ICAM, NFKB1 and CXCL2 . It was concluded that while the MW fractionation potentially generated compounds displaying distinct immune modulatory functions, the crude EE exhibited the most potent immunomodulatory bioactivity. Industrial relevance The result demonstrated that the fractionation of crude 80% ethanol extract of A. nodosum into MW fractions potentially separate bioactive compounds with distinct immune modulatory function. The crude extract exhibited the most potent immunomodulatory bioactivity in Caco-2 cells challenged with TNF-α and in the porcine colonic tissue challenged with LPS. Such a broad spectrum anti-inflammatory bioactivity of A. nodosum ethanol extract can be further explored for its potential application in inflammatory diseases of the mammalian intestine.

Summary The technique of challenging postmortem tissue explants with inflammation inducer such as lipopolysaccharide (LPS) followed by gene expression analysis is used widely for evaluating the immune-suppressing effect of bioactives. Using porcine colonic tissue as an ex-vivo model of mammalian intestinal gut, this study evaluated the effect of incubation time on the integrity of gene transcripts and activation of inflammatory immune gene cascade by LPS treatment. Post-slaughter colon was removed surgically and explants were incubated for 0, 3, 6 and 12 h and the abundance of mRNA transcripts of a panel of 92 immune genes were evaluated using quantitative polymerase chain reaction (qPCR) arrays. The mRNA transcripts were highly intact after 0 and 3 h of incubation; however, after 6 h the degradation was clearly evident. Following 3 h incubation, 98·8% and 100% mRNA transcripts were detectable in the colonic tissue harvested from weaned and mature pigs, respectively. In the explants of weaned piglets, LPS treatment activated inflammatory signalling pathways [high mobility group B1 (HMGB1), dendritic cell maturation, interleukin (IL)-6, IL-8, IL-17F], while these pathways were inhibited by dexamethasone treatment. Activations of inflammatory genes were also evident in the explants collected from the mature pigs subjected to ex-vivo incubation for 3 h in the absence or presence of LPS. It is concluded that the colonic explant remains physiologically viable and responsive to immunological challenge for up to 3 h ex-vivo.

Adenosine receptor–mediated regulation of monocyte/macrophage inflammatory responses is critical in the maintenance of tissue homeostasis. In this study, we reveal that adenosine potently modulates the expression of NR4A1, 2, and 3 orphan nuclear receptors in myeloid cells, and this modulation is primarily through the adenosine A2a receptor subtype. We demonstrate that A2a receptor activation of NR4A1-3 receptor synthesis is further enhanced in TLR4-stimulated monocytes. After TLR4 stimulation, NR4A receptor–depleted monocyte/macrophage cells display significantly altered expression of cell-surface markers and produce increased inflammatory cytokine and chemokine secretion rendering the cells an enhanced proinflammatory phenotype. Exposure of TLR4 or TNF-α–stimulated monocytes to adenosine analogs directs changes in the expression of MIP-3α and IL-23p19, with NR4A2 depletion leading to significantly enhanced expression of these factors. Furthermore, we establish that nuclear levels of NF-κB/p65 are increased in TLR/adenosine-stimulated NR4A2-depleted cells. We show that, after TLR/adenosine receptor stimulation, NR4A2 depletion promotes significant binding of NF-κB/p65 to a κB consensus binding motif within the MIP-3α proximal promoter leading to increased protein secretion, confirming a pivotal role for NF-κB activity in controlling cellular responses and gene expression outcomes in response to these mediators. Thus, these data demonstrate that during an inflammatory response, adenosine modulation of NR4A receptor activity acts to limit NF-κB–mediated effects and that loss of NR4A2 expression leads to enhanced NF-κB activity and hyperinflammatory responses in myeloid cells.

Dysregulation of inflammatory responses is a hallmark of multiple diseases such as atherosclerosis and rheumatoid arthritis. As constitutively active transcription factors, NR4A nuclear receptors function to control the magnitude of inflammatory responses and in chronic inflammatory disease can be protective or pathogenic. Within this study, we demonstrate that TLR4 stimulation using the endotoxin lipopolysaccharide (LPS) rapidly enhances NR4A1–3 expression in human and murine, primary and immortalized myeloid cells with concomitant gene transcription and protein secretion of MIP-3α, a central chemokine implicated in numerous pathologies. Deficiency of NR4A2 and NR4A3 in human and murine myeloid cells reveals that both receptors function as positive regulators of enhanced MIP-3α expression. In contrast, within the same cell types and conditions, altered NR4A activity leads to suppression of LPS-induced MCP-1 gene and protein expression. An equivalent pattern of inflammatory gene regulation is replicated in TNFα-treated myeloid cells. We show that NF-κB is the critical regulator of NR4A1–3, MIP-3α, and MCP-1 during TLR4 stimulation in myeloid cells and highlight a parallel mechanism whereby NR4A activity can repress or enhance NF-κB target gene expression simultaneously. Mechanistic insight reveals that NR4A2 does not require DNA-binding capacity in order to enhance or repress NF-κB target gene expression simultaneously and establishes a role for NF-κB family member Relb as a novel NR4A target gene involved in the positive regulation of MIP-3α. Thus, our data reveal a dynamic role for NR4A receptors concurrently enhancing and repressing NF-κB activity in myeloid cells leading to altered transcription of key inflammatory mediators.

Active pulmonary tuberculosis (APTB) is associated with a failure of the host immune system to control the invading Mycobacterium tuberculosis (Mtb). The objective of this study was to quantify and assess the role of serum inflammatory cytokines in active pulmonary tuberculosis patients following anti-tuberculosis drug (ATD) therapy. Blood samples were collected from APTB patients and normal healthy subjects (NHS) (total n = 204) at baseline and 2, 4 and 6 months post-therapy and the abundance of serum inflammatory cytokines were measured by cytokine specific ELISA. Compared to NHS, APTB patients at baseline had higher levels of serum pro-inflammatory cytokines IL-12p40 (P < 0.001), IFN-γ (P < 0.001), TNF-α (P < 0.01), IL-1β (P < 0.001) and IL-6 (P < 0.001) and anti-inflammatory cytokines IL-10 (P < 0.001) and TGF-β1 (P < 0.001) while there was no change in the level of IL-4. In APTB patients, the serum levels of IFN-γ, TNF-α, IL-6 and TGF-β1 directly relate to the bacterial load while the TNF-α, IL-1β, IL-6 and TGF-β1 relate to radiological severity. At baseline, the IL-6 level in NHS and APTB patients differed most and following ATD therapy, this level rapidly decreased and stabilized by 4-month in APTB patients. It is concluded that a subtle reduction in the serum level of IL-6 of the APTB patients following ATD therapy might play a vital role in immune-protection of the host against Mtb infection and hence the serum IL-6 level can be a useful marker to diagnose the effectiveness of therapy in the patients.

Bioactive milk peptides are reported to illicit a range of physiological benefits and have been proposed as potential functional food ingredients. The objective of this study was to characterize the anti-inflammatory properties of sodium caseinate (NaCAS), its enzyme hydrolysate (EH) and peptide-enriched fractions (5 kDa retentate [R], 1 kDaR and 1 kDa permeate [P]), both in vitro using a Caco-2 cell line, and also ex vivo using a porcine colonic tissue explant system. Caco-2 cells were stimulated with tumour necrosis factor alpha (TNFα) and co-treated with casein hydrolysates for 24 h. Following this, interleukin (IL)-8 concentrations in the supernatant were measured using enzyme-linked immunosorbent assay. Porcine colonic tissue was stimulated with lipopolysaccharide and co-treated with casein hydrolysates for 3 h. The expression of a panel of inflammatory cytokines was measured using qPCR. While dexamethasone reduced the IL-8 concentration by 41.6%, the 1 kDaR and 1 kDaP fractions reduced IL-8 by 68.7% and 66.1%, respectively, relative to TNFα-stimulated Caco-2 cells (P < 0.05). In the ex vivo system, only the 1 kDaR fraction elicited a decrease in IL1-α, IL1-β, IL-8, TGF-β and IL-10 expression (P < 0.05). This study provides evidence that the bioactive peptides present in the 1 kDaR fraction of the NaCAS hydrolysate possess anti-inflammatory properties in vitro and ex vivo. Further in vivo analysis of the anti-inflammatory properties of the 1 kDaR is proposed.

RATIONALE Isotope ratio analysis of bovine tissues is a tool for inferring aspects of the dietary history of cattle. The objective of this experiment was to quantify the carbon (C) and nitrogen (N) isotopic turnover in blood (serum and residue) and inner organs (liver, kidney, heart and brain) of beef cattle. METHODS Growing beef cattle (n = 70 in total) were either switched from a control diet containing barley and urea to an experimental diet containing maize and 15N-enriched urea, for various intervals prior to slaughter or maintained on the control diet for 168 days pre-slaughter. Samples of blood, liver, kidney, heart and brain were collected at 0, 14, 28, 56, 112 and 168 days and analysed using Isotope Ratio Mass Spectrometry. RESULTS After 168 days, C- and N-isotopic equilibrium was reached in the blood serum, liver and kidney, approached in the heart and brain, but not reached in the non-serum component of blood. The estimated C and N half-lives were 16.5 and 20.7 days for liver, 19.2 and 25.5 days for kidney, 29.2 and 35.6 days for blood serum, 37.6 and 49.9 days for heart, 53.3 and 52.2 days for brain and 113.3 and 115.0 days for the non-serum blood residue, respectively. Modelling the C and N turnover in the different tissue combinations revealed that a combined analysis of liver and heart as well as brain and kidney can provide the most accurate estimation of the timing of the diet switch. CONCLUSIONS Based on the difference in turnover rates, bovine soft tissues can provide isotopic information on short- and long-term dietary changes, which in turn may be linked to the geographic or production origin of beef cattle. This study also provides basic biological data on organ C and N turnover in a large herbivorous mammal. Copyright © 2014 John Wiley & Sons, Ltd.

Citrus peel, a by-product of citrus processing is a rich source of numerous bioactive nutrients. This experiment was conducted to evaluate the effects of dietary supplementation of Citrus sinensis peel extract (CSPE) on few production and quality traits of broiler chickens. Four hundred broiler chicks (one-day-old) were assigned to any of the five dietary treatments comprised of: basal diet for 42 days, or basal diet containing 1000 ppm or 1250 ppm CSPE for 21 days, or basal diet containing 1000 ppm or 1250 ppm CSPE for 42 days. Compared to the final weight of broilers on the basal diet (2426.3 ± 120.9 g), the supplementation of the diet with 1000 ppm CSPE for 21 days resulted in significantly (P < 0.05) higher final weight (2692.5 ± 120.9 g), while that with 1250 ppm CSPE for 42 days resulted in a significantly lower (P < 0.05) abdominal fat content (47.94 vs. 28.75 ± 4.38 g). Supplementation with 1000 or 1250 ppm CSPE for 21 days or 42 days had no significant effect on the final weight, empty body weigh...

An experiment (complete randomised design) was conducted to investigate the effects of supplementing different molecular weights (MW) of chitooligosaccharide (COS) on pig performance, selected microbial populations and nutrient digestibility post-weaning. A total of 396 weaned piglets (24 days of age, 7.3 kg ± (s.d.) 1.7 kg live weight) were assigned to one of six dietary treatments (22 replicates/treatment) for a 33-day experimental period. The dietary treatments were as follows (1) control diet (0 ppm COS), (2) control diet plus 0.05). However, there were quadratic responses in both daily gain (P < 0.05) and gain to feed ratio (P < 0.05) to the increased MW of COS inclusion during the weaner period (days 18 to 33) with all COS-supplemented treatments improving daily gain and gain to feed ratio compared with the control. There was a quadratic response in faecal scoring to the increased MW of COS inclusion from days 0 to 7 (P < 0.001), days 7 to 14 (P < 0.001) and during the overall experimental period (P < 0.01) with all the COS-supplemented treatments having an improved faecal score compared with the control. During the weaner period, there was a cubic response in lactic acid bacteria and Escherichia coli populations as the MW of COS increased (P < 0.05). The 5 to 10 kDa and 10 to 50 kDa COS increased lactic acid bacteria populations compared with the control, whereas lactic acid bacteria populations decreased at 50 to 100 kDa. The 5 to 10 kDa, 10 to 50 kDa and 50 to 100 kDa COS decreased E. coli populations compared with the control. There was a cubic response in the apparent total tract digestibility of dry matter (DM; P < 0.01), organic matter (OM; P < 0.01), ash (P < 0.01), nitrogen (N; P < 0.01) and gross energy (GE; P < 0.01) to the increased MW of COS inclusion during the weaner period. The 5 to 10 kDa COS had a higher apparent total tract digestibility of DM, OM, ash, N and GE in comparison to the control, whereas the apparent total tract nutrient digestibility of these nutrients decreased at 10 to 50 kDa. The current results indicate that the MW ranges of 5 to 10 kDa and 10 to 50 kDa COS decreased E. coli numbers while increasing nutrient digestibility of the diets.

Conventional dietary strategies to reduce pig manure odor may either be costly, or impede nutrient digestibility. Additionally, the response of manure odor to such measures may be variable, indicating a complex relationship between environmental pollutant and diet. We hypothesized that dietary Lactobacillus plantarum (LP), with or without the inclusion of a purified oligofructose (inulin), may reduce odor without compromising nutrient digestibility. An experiment with a 2 × 2 factorial arrangement of treatments was conducted to investigate effects of dietary inulin (0 and 12.5 g/kg) and LP (0 and 0.5 g/kg) on nutrient digestibility, indicators of gastrointestinal tract fermentation, select fecal bacteria, manure content, and ammonia and odor emissions of 28 growing-finishing pigs (60.3 kg; n = 7/treatment). Dietary treatments had no effect on nutrient digestibility. Dietary treatments containing inulin had decreased Enterobacteriaceae (8.60 vs. 9.67 log gene copy number/g fresh feces; P = 0.03) when compared with unsupplemented diets. There was an interaction between dietary inulin concentration and LP supplementation on estimates of fecal Clostridia (P = 0.01). Pigs offered diets containing both inulin and LP in combination had increased Clostridia when compared with those offered the control diet. However, there was no effect of either LP or inulin fecal Clostridia when offered singularly. An interaction was also noted where diets supplemented with LP or inulin only reduced odor (P = 0.01) compared with the control diet. However, there was no effect of LP on manure odor emissions when offered in combination with inulin. In summary, this study demonstrated that dietary supplementation with either exogenous LP or inulin reduces manure odor but not when offered in combination.

The present study was conducted to investigate the effect of maternal dietary supplementation (n = 10 sows/treatment) with seaweed extract (SWE: 0 vs. 10.0 g/d) from d 107 of gestation until weaning (d 26) on neonatal piglet growth, humoral immunity, intestinal morphology, selected intestinal microflora, and VFA concentrations. Furthermore, this study examined the effect of dietary treatment on the immune response after an ex vivo Escherichia coli lipopolysaccharide (LPS) tissue challenge at weaning in a 2 × 2 factorial arrangement. The main factors consisted of sow dietary treatment (SWE or control) and immunological challenge (yes or no). The SWE supplement (10.0 g/d) contained laminarin (1.0 g), fucoidan (0.8 g), and ash (8.2 g) and was extracted from a Laminaria spp. The SWE-supplemented sows had greater colostrum IgA (P < 0.01) and had a trend for greater IgG (P = 0.062) concentrations compared with non-SWE-supplemented sows. Piglets suckling SWE-supplemented sows had greater serum IgG (P < 0.05) concentrations on d 14 of lactation compared with those suckling non-SWE-supplemented sows. Dietary SWE supplementation decreased fecal Enterobacteriaceae populations in sows at parturition (P < 0.05), and piglets suckling SWE-supplemented sows had a decreased colonic E. coli population at weaning (P < 0.01) compared with non-SWE-supplemented sows. Lipopolysaccharide challenge increased the mRNA abundances of the pro-inflammatory cytokines IL-1α and IL-6 (P < 0.01) in ileal tissue and tumor necrosis factor (TNF)-α in colonic (P < 0.01) tissue. There was a treatment × LPS challenge interaction for ileal TNF-α mRNA expression (P < 0.05). Piglets suckling SWE-supplemented sows had greater TNF-α mRNA expression after ex vivo LPS challenge compared with non-SWE-supplemented sows (P < 0.05). However, there was no effect of sow dietary treatment on TNF-α mRNA expression in the unchallenged ileal tissue. Piglet BW at birth and weaning, and small intestinal morphology were unaffected by sow dietary treatment under current experimental conditions. In summary, these results demonstrate an important immunomodulatory role of SWE supplementation characterized by enhanced colostral IgA and IgG concentrations, greater piglet circulatory IgG concentrations on d 14 of lactation, and enhanced TNF-α mRNA expression in the ileum after an ex vivo LPS challenge. These results indicate that SWE supplementation enhanced piglet immune function and colonic microflora at weaning.

An experiment (complete randomised design) was conducted to investigate the effects of supplementing different molecular weights (MW) of chitooligosaccharide (COS) on intestinal morphology, selected microbial populations, volatile fatty acid (VFA) concentrations and the immune status of the weaned pig. A total of 28 piglets (24 days of age, 9.1 kg (± s.d. 0.80) live weight) were assigned to one of four dietary treatments for 8 days and then sacrificed. The treatments were (1) control diet (0 ppm COS), (2) control diet plus 5 to 10 kDa COS, (3) control diet plus 10 to 50 kDa COS and (4) control diet plus 50 to 100 kDa COS. The COS was included in dietary treatments at a rate of 250 mg/kg. Tissue samples were taken from the duodenum, jejunum and ileum for morphological measurements. Digesta samples were taken from the proximal colon to measure lactobacilli and Escherichia coli populations and digesta samples were taken from the caecum and proximal colon for VFA analysis. Gene expression levels for specific cytokines were investigated in colonic tissue of the pig. Supplementation of different MW of COS had no significant effect on pig performance during the post-weaning period (days 0 to 8; P > 0.05). The inclusion of COS at all MW in the diet significantly reduced faecal scores compared with the control treatment (P < 0.01). Pigs fed the 10 to 50 kDa COS had a higher villous height (P < 0.05) and villous height : crypt depth ratio (P < 0.05) in the duodenum and the jejunum compared with the control treatment. Pigs fed the 5 to 10 kDa COS had a lower lactobacilli population (P < 0.05) and E. coli population (P < 0.05) in the colon compared with the control group. Pigs offered the 5 to 10 kDa COS had significantly lower levels of acetic acid and valeric acid compared with the control group (P < 0.05). The inclusion of different MW of COS had no significant effect on the expression of the cytokines tumour necrosis factor-α, Interleukin (IL)-6, IL-8 and IL-10 in the gastro-intestinal tract of the weaned pig. The current results indicate that a lower MW of 5 to 10 kDa COS possessed an antibacterial activity, while the higher MW of 10 to 50 kDa was optimum for enhancing the intestinal structure.

The neuropeptide Y 5 receptor (NPY5R) plays an important role in the regulation of appetite and feeding behaviour in mammals by modulating the effect of the neurotransmitter neuropeptide Y. As single nucleotide polymorphism (SNP) variation in the bovine NPY5R gene is likely to influence the expression and/or function of this gene, the objectives of this study were to identify SNPs in the bovine NPY5R gene and to predict their functional role in the expression and physico–chemical characteristics of the protein product. Nineteen novel SNPs were identified in a 2.1 kb genomic region of the NPY5R gene in a total of 419 beef cattle from 13 Bos taurus breeds and eight Bos indicus animals. Four of these SNPs were non-synonymous (Met → Ile, Leu → Phe, Pro → Leu, Arg → Stop codon), while 10 were synonymous. Of particular interest was one non-synonymous SNP (c.1090C>T) that introduced a stop codon in the third intracellular loop of the NPY5R molecule. This stop codon is predicted to create a truncated NPY5R molecule with different physico–chemical properties compared to the native NPY5R protein. A further four SNPs were located in the 5′ untranslated region (UTR) and one in the 3′UTR. Two of the 5′UTR SNPs affected putative transcription factor binding sites (GATA binding factor and snRNA-activating protein complex). In conclusion, regulatory and functional SNPs were identified in the bovine NPY5R gene. These include SNPs which potentially modify transcription factor binding sites as well as SNPs that cause amino acid changes and premature termination of the NPY5R protein. Such polymorphisms are likely to play vital physiological roles in the neuropeptide Y mediated appetite, feed intake and energy homeostasis in cattle.

Neuropeptide Y (NPY) is a potent orexigenic agent. The molecular mechanisms underlying the regulation of bovine NPY gene expression by its promoter region is currently unknown. The objectives of this research were to: (i) identify the SNPs in the promoter region of the bovine NPY gene, (ii) investigate the effects of these SNPs by measuring promoter transcriptional activities of different bovine NPY promoter haplotypes and; (iii) identify the minimal promoter region (MPR) required for basal activity of the NPY gene in vitro. Seventeen SNPs were identified in the promoter region. Of these, 14 affected putative transcription factors binding motifs including a TATA binding protein factor at -20, GC-Box factors SP1 at -170 and GATA binding motifs at -120 and -347. The SNPs were assigned to five major haplotypes (BtNPY_H1-5), of which BtNPY_H5 had maximum transcriptional activity. The region extending to -134 nt was identified as the MPR. This MPR was confirmed by the identification of a putative TATA box (-29 nt) and two SP1/GC binding sites (-94 and -118 nt), within this region. However, promoter expression was significantly enhanced when the construct contained the -614 to -1019 nt region. In conclusion, a number of SNPs characterised in the bovine NPY promoter especially those affecting the transcription factor binding sites, enhancer and repressor regions have the potential to affect NPY gene expression. Natural variation exists in the promoter region of the bovine NPY gene, which should be further explored for selection of energetic efficiency in cattle.

The present study investigated the effects of dietary supplementation of a seaweed extract (SWE) to sows and weaned pigs on post-weaning growth performance, intestinal morphology, intestinal microflora, volatile fatty acid concentrations and immune status of pigs at days 11 and 117 post-weaning. Gestating sows (n20) were supplemented with a SWE (0v.10·0 g/d) from day 107 of gestation until weaning (day 26). At weaning, pigs (four pigs per sow) were divided into two groups based on sow diet during lactation and supplemented with a SWE (0v.2·8 g/kg diet), resulting in four treatment groups: (1) BB (basal sows–basal pigs); (2) BS (basal sows–treated pigs); (3) SB (treated sows–basal pigs); (4) SS (treated sows–treated pigs). Pigs weaned from SWE-supplemented sows had a higher average daily gain (ADG) between days 0 and 21 (P Escherichia colipopulations on day 11 (P P MUC2in the colon at day 11 post-weaning (P

Genetic polymorphisms present in the bovine lactoferrin (LTF) gene promoter have the potential to affect milk lactoferrin concentrations. The objectives were: (1) to identify, in silico, SNPs in the promoter region of the LTF gene that could affect transcription factor binding activity, (2) to investigate the effects of these SNPs in vitro by measuring promoter transcriptional activities of different bovine LTF promoter haplotypes and (3) to investigate the genetic association between LTF promoter SNPs and milk lactoferrin concentration. Haplotypes were deduced from sequencing of the 2.2-kb bovine LTF promoter in 78 unrelated animals. In silico analysis of the 2.2-kb promoter revealed two major haplotypes (BtLTF_H1a and BtLTF_H2a) that differed at 10 SNP loci that affect transcription factors of both a constitutive (at -28, -1702) and an inducible (at -131, -270, -586, -2047, -2077, -2122, -2140 and -2151) nature. The basal promoter transcriptional activity of BtLTF_H1a was 1.44-fold higher than that of BtLTF_H2a in mammary epithelial cells. Cows with the BtLTF_H1a haplotype had increased lactoferrin protein concentration in milk at various time points over the lactation curves, compared to herdmates with the BtLTF_H2a haplotype. The SNPs c.-28A>C, c.-131T>C, c.-156A>G, c.-270T>C, c.-586C>T, c.-1702A>G, c.-1953G>A, c.-2047A>G, c.-2077A>G, c.-2122C>T, c.-2140A>G and c.-2151G>A were associated (P < 0.001) with milk lactoferrin content in 372 Holstein-Friesian cows. The identification of bovine LTF promoter haplotypes with different basal transcriptional activities in vitro that are associated with lactoferrin levels in milk in vivo may facilitate the identification of designer dairy herds for increased lactoferrin content in milk.

A 2 × 2 factorial study (n = 10 sows/treatment) was performed to investigate the effects of maternal dietary supplementation with seaweed extract (SWE) (yes vs. no) and fish oil (FO) inclusion (0 vs. 100 g/day) from day 109 of gestation until weaning (24 days) on piglet performance and intestinal microflora, intestinal morphology and inflammatory gene expression in 9 day old weaned piglets. There was an interaction between SWE and FO on E. coli populations (P

An experiment with a 2 x 2 factorial arrangement of treatments (n = 10 sows/treatment) was conducted to investigate the effect of maternal dietary supplementation with seaweed extract (SWE: 0 vs. 10.0 g/d) and fish oil (FO) inclusion (0 vs. 100 g/d) from d 109 of gestation until weaning (d 26) on sow colostrum and milk composition, humoral immune response on d 5 and 12 of lactation, and suckling piglet performance. Furthermore, the influence of dietary treatment on the phagocytic activity of whole blood white cells at weaning was examined. The SWE (10 g) contained laminarin (1 g), fucoidan (0.8 g), and ash (8.2 g) and was extracted from a Laminaria spp. The FO contained approximately 40% eicosapentaenoic acid and 25% docosahexaenoic acid. The SWE-supplemented sows had greater colostrum IgG (P < 0.01) and milk protein (P < 0.05) concentrations on d 12 of lactation compared with non-SWE-supplemented sows. Piglets suckling SWE-supplemented sows had greater serum IgG (P < 0.01) and IgA (P < 0.05) concentrations on d 5 and IgG concentrations on d 12 (P < 0.05) of lactation compared with those suckling non SWE-supplemented sows. In contrast, FO supplementation exerted a suppressive effect on piglet serum IgA concentrations on d 5 of lactation (P < 0.05) compared with non-FO-supplemented diets. Dietary FO supplementation enhanced the n-3 PUFA proportion of sow milk (P < 0.001) and piglet serum at weaning (P < 0.001). Piglets suckling SWE-supplemented sows had a greater percentage of Escherichia coli phagocytizing leukocytes (P < 0.05) and a reduced percentage of E. coli phagocytizing lymphocytes (P < 0.01) compared with non-SWE-supplemented sows. Piglets suckling FO-supplemented sows had a greater percentage of leukocytes (P < 0.05) and lymphocytes (P < 0.05) phagocytizing E. coli compared with non-FO-supplemented sows. However, total leukocyte, lymphocyte, monocyte, and neutrophil numbers were not influenced by sow dietary treatment. Average piglet weaning weight and ADG between birth and weaning were not influenced by sow dietary treatment. In conclusion, the current study demonstrates that SWE supplementation from d 109 of gestation until weaning enhanced colostral IgG concentrations and circulatory IgG concentrations in suckled piglets on d 5 and 12 of lactation. Furthermore, the percentage of leukocytes and lymphocytes phagocytizing E. coli at weaning increased in piglets suckling FO-supplemented sows, indicating an enhancement of immune function against presenting pathogens. However, the combination of SWE and FO bestowed no positive effect on immune responses investigated in the current study.

A 2 × 2 factorial experiment (n = 10 sows/treatment) was performed to investigate the effects of maternal dietary supplementation with seaweed extract (SWE) (yes vs. no) and fish oil (FO) inclusion (0 vs. 100 g/day) from day 109 of gestation on colostrum composition, colostrum immunoglobulin concentrations, humoral immunity and growth of the suckling piglets. The SWE contained laminarin and fucoidan while the FO contained 40% eicosapentaenoic acid (EPA) and 25% docosahexaenoic acid (DHA). In the piglets suckling the SWE supplemented sows, the concentrations of serum immunoglobulin (Ig) G were higher on days 5 (P

Lactoferrin plays an important role in the innate immune system, with well-characterized antibacterial, antiviral, and immune modulatory properties. The objective of this study was to determine the allele and haplotype frequency of polymorphisms at positions −586, −190, and −28 of the bovine lactoferrin promoter in Holstein-Friesians and to quantify their association with performance using phenotypic data on progeny from 848 sires. Associations between genotypes and performance were quantified using weighted mixed models with genotyped individuals included as a random effect, and average expected relationships among individuals accounted for through a numerator relationship matrix. The dependent variables were daughter yield deviation for production traits and deregressed predicted transmitting ability for calving interval and functional survival. The C to T polymorphism at −586, which distorts a putative activating protein 2 (AP-2) binding site, was associated with a shorter calving interval and higher somatic cell score. The G to A polymorphism at −190, located in a putative selective promoter factor 1 (SP-1) binding site, was associated with a longer calving interval and decreased functional survival. A third polymorphism (A to C) at position −28, found within the noncanonical TATA box, had a tendency to associate with functional survival. On the basis of the data we proposed a haplotype combination that was associated with improved reproductive performance in the Holstein-Friesian breed. We hypothesized that the observable phenotypic associations with lactoferrin promoter polymorphisms can potentially be explained by allele-specific differences in constitutive or inducible levels of gene expression. The lack of a pleiotropic effect of the single nucleotide polymorphisms studied on both fertility and milk production traits strengthens the importance of these polymorphisms, or at least the lactoferrin promoter, in selection for improved fertility.

Stable isotope ratio analysis of light elements (including C, N, and S) is a powerful tool for inferring the production and geographic origins of animals. The objectives of this research were to quantify experimentally the isotopic turnover of C, N, and S in bovine skeletal muscle (LM and psoas major) and to assess the implications of the turnover for meat authentication. The diets of groups (n = 10 each) of beef cattle were switched from a control diet containing barley and unlabelled urea to an experimental diet containing maize, (15)N-labeled urea, and seaweed for periods of up to 168 d preslaughter. The feeding of the experimental diet was clearly reflected by the delta(13)C, delta(15)N, and delta(34)S values of the LM and psoas major muscles, but isotopic equilibrium was not reached in either muscle for C, N, or S after 168 d of feeding the experimental diet. The slow turnover in skeletal muscle was reflected by the C and N half-lives of 151 and 157 d for LM and 134 and 145 d for psoas major, respectively, and by an S half-life of 219 d in LM. It is concluded that the turnover of light elements (C, N, and S) in bovine skeletal muscles is a slow process; therefore, skeletal muscles contain isotopic information on dietary inputs integrated over a long period of time (months to years).