Incidences of aflatoxin contaminations in ingredients, feed and products of poultry from two regions in Ghana.

Commercial poultry feed required for proper growth of birds and egg production contains essential nutrients with maize as key component. Inadequately dried maize is prone to aflatoxin contamination and therefore when used in feed formulation for poultry may compromise the safety of the feeds and poultry products. This study investigated the levels of aflatoxins in feed ingredients, feed and poultry products sampled from Eastern and Greater -Accra regions of Ghana. The aflatoxin levels of B1, B2, G1, and G2 were determined using a High-Performance Liquid Chromatography (HPLC) methodology. Main feed ingredients used were fishmeal, cotton seed cake, soya meal, rice, wheat and maize bran as well as maize grains. There was correlation between the level of aflatoxins and moisture content in poultry feed ingredients. All the poultry feeds (100 %) analysed showed the presence of aflatoxins with total aflatoxins recorded ranging from 5.32 to 29.88 µg/kg. Maize samples of the poultry feeds, from all two regions, revealed maize to be a major contributor to the overall total aflatoxin contents found in the feed. Five (5) out of ten (10) communities investigated in the two (2) regions where the poultry feeds were examined recorded total aflatoxin levels in maize above the Ghana Standard Authority (GSA) specification of 20 µg/kg. Aflatoxins G1 and G2 were not detected in all samples of chicken meat and eggs. Total aflatoxin levels recorded for all chicken meat and eggs were below GSA specification of 5 µg/kg; implying that these products were safe for consumption.

Intravaginal injection of Lactobacillus johnsonii may modulates oviductal microbiota and mucosal barrier function of laying hens.

The avian oviduct connects to the gastrointestinal tract through cloaca, where it is exposed to pathogenic bacteria from intestinal contents. Therefore, improvement of mucosal barrier function in the oviduct is important for safe poultry production. Lactic acid bacteria are known to contribute to strengthening the mucosal barrier function in the intestinal tract, and a similar effect is expected in the oviduct mucosa of chickens. This study aimed to clarify the effects of vaginal administration of lactic acid bacteria on the mucosal barrier function of the oviduct. White Leghorn laying hens (500-days old) were intravaginally administered 1 mL of Lactobacillus johnsonii suspension (1 × 105 and 1 × 108 cfu/mL: low concentration of Lactobacillus (LL) and high concentration of Lactobacillus (HL) groups, respectively) or without bacteria (control: C group) for 7 d (n = 6). The oviductal magnum, uterus, and vagina were collected for histological observations and mucosal barrier function-related gene expression analysis. Amplicon sequence analysis of oviductal mucus bacteria was also performed. Eggs were collected during the experimental period and their weight was measured. Vaginally administering L. johnsonii for 7 d caused 1) an increase in α-diversity of vaginal mucosa microbiota with an increase in the abundance ratio of beneficial bacteria and a decrease in pathogenic bacteria, 2) enhanced claudin (CLA) 1 and 3 gene expression in the magnum and vaginal mucosa, and 3) a decrease in avian ß-defensin (AvBD) 10, 11, and 12 gene expression in the magnum, uterus, and vaginal mucosa. These results suggest that transvaginal administration of L. johnsonii contributes to protection against infection in the oviduct by improving the microflora of the oviductal mucosa and strengthening the mechanical barrier function of the tight junctions. In contrast, transvaginal administration of lactic acid bacteria does not enhance the production of AvBD10, 11, and 12 in the oviduct.

Heavy metal, microbial and pesticides residue contaminations are limiting the potential consumption of green leafy vegetables in Ghana: An overview.

Green leafy vegetables (such as cocoyam (Colocasia spp) leaves, spinach (Spinach spp), amaranths (Amaranthus spp), roselle leaves (Hibiscus spp), and lettuce (Lactuca spp)) form a major part of Ghanaian meals providing essential vitamin such as A, B and C and minerals including iron and calcium as well as essential bioactive compounds. However, the practices involved in the production, distribution and handling of these nutrient rich vegetables, by most value chain actors in Ghana, unfortunately pre-dispose them to contamination with pathogens, heavy metals and pesticides residues. These have therefore raised public health concerns regarding the safety and quality of these green leafy vegetables. Understanding the current perspectives of the type of pathogens, heavy metals and pesticide contaminants that are found in leafy vegetables and their health impacts on consumers will go a long way in helping to identify appropriate mitigation measures that could be used to improve the practices involved and thereby help safeguard human health. This review examined reported cases of microbial, heavy metal and pesticides residue contamination of green leafy vegetables in Ghana from 2005 to 2022. Notable pathogenic microorganisms were Ascaris eggs and larvae, faecal coliform, Salmonella spp., Staphylococcus aureus Streptococci, Clostridium perfringes, and Escherichia coli. In addition, Lead (Pb), Cadmium (Cr), Chromium (Cr), Zinc (Zn), Iron (Fe), Copper (Cu) and Manganese (Mn) have been detected in green leafy vegetables over the years in most Ghanaian cities. Pesticides residues from organochlorine, organophosphorus and synthetic pyrethroid have also been reported. Overall, microbial, heavy metals and pesticide residue contamination of Ghanaian green leafy vegetables on the farms and markets were significant. Hence, mitigation measures to curb the contamination of these vegetables, through the food chain, is urgently required to safeguard public health.

Aging-associated increased nitric oxide production is a potential cause of inferior eggshell quality produced by aged laying hens.

It is important to prolong the productive life of laying hens without compromising their welfare. Therefore, in this study, we aimed to identify the cause for inferior quality egg production of aged hens by investigating the aging-associated molecular changes related to eggshell formation in the isthmic and uterine mucosae and determining whether nitric oxide plays a role in decreasing the quality of eggs produced by aged hens. Young (35 weeks old) and aged (130 weeks old) White Leghorn laying hens were used in this study to determine the effects of age on the expression of proteins related to eggshell membranes formation in the isthmus and eggshell biomineralization and nitric oxide production in the uterus. Nitric oxide synthesis during the ovulatory cycle was examined in twenty-five laying hens (46-52 weeks old) euthanized at 0, 4, 7, 16, and 24 h after oviposition. S-Nitroso-N-acetylpenicillamine (a nitric oxide donor) was added to the cultured isthmic and uterine mucosal cells to examine the effects of nitric oxide on the expression of genes related to eggshell membranes formation and eggshell biomineralization, respectively. The results showed that the protein abundance of collagen I and V in the isthmic mucosa and collagen V in the eggshell membranes were lower in aged hens than in young hens. The mRNA expression levels of calbindin, osteopontin, and ovocalyxin-36 and the protein abundance of calbindin and carbonic anhydrase-2 were lower in the uterine mucosa of aged hens than in that of young hens. Nitric oxide synthesis was higher in the uterine mucosa of aged hens than in that of young hens. Nitric oxide downregulated the mRNA expression levels of osteopontin and ovocalyxin-36 in cultured uterine mucosal cells. Our results indicated that the eggshell quality decreases with aging due to molecular changes in the uterine mucosa affecting the eggshell membrane formation and eggshell biomineralization. Moreover, nitric oxide overproduction may play a role in this dysfunction.

Effects of two pre-treatments, blanching and soaking, as processing modulation on non-enzymatic browning developments in three yam cultivars from Ghana.

Non-enzymatic browning develops in dry-cooked foods and those with high carbohydrate develop acrylamide, a neurotoxin and potential carcinogen. However, some non-enzymatic browning products have reducing properties. We hypothesized that non-enzymatic browning and reducing power, a measure of antioxidant activity, of processed yam are affected by pre-treatment. Peeled yam cultivars (KM, RKD and SO89) in chunks were pre-soaked (0, 3, 6, 12, 18, and 24 h) in distilled water or pre-blanched (0, 1, 2, 3, 4, and 5 min) in steam. Pre-treated samples were deep-fried at 180 °C for 15 min or roasted at 220 °C for 30 min. Soluble solids, titratable acidity and pH of yam tissues and soaking water were determined. pH of the soaked yam tissues showed a positive relation with non-enzymatic browning. Pre-soaked fried KM and roasted RKD showed a significant decrease in non-enzymatic browning intensities. The reducing power of the cooked yams ranged between 78.94 and 185.92 % of ascorbic acid, and was affected by the different pre-treatment and dry-cooking methods.

Effect of antibiotic treatment on microbial composition and expression of antimicrobial peptides and cytokines in the chick cecum.

The aim of this study was to confirm whether the expression of innate immune molecules in the chick cecum is altered in association with changes in the composition of the intestinal microbiome that are regulated by treatment with antibiotics. Broiler chicks were administered with antibiotics (penicillin and streptomycin) daily, and the composition of the microbiota, expression of innate immune molecules, and localization of antimicrobial peptides in the chick cecum were examined at day 7 and day 14 using real-time PCR and immunohistochemistry. The oral administration of antibiotics caused an increase in the relative frequency of the Enterobacteriaceae family and a decrease in some gram-negative (Barnesiellaceae) and gram-positive bacterial (Clostridiaceae and Erysipelotrichaceae) families. The gene expression levels of immune molecules, including 4 Toll-like receptors (TLR) (TLR 2, 4, 5, and 21), inflammation-related cytokines (IL-1ß, TGFß3, TGFß4, and IL-8), and antimicrobial peptides (avian ß-defensins and cathelicidins) showed a tendency to decrease with antibiotic treatment at day 7. However, expression levels of TLR21 and some cytokines (IL-1ß, TGFß3, and IL-8) were higher in the cecum or cecal tonsils of the antibiotic-treated group than in those of the control at day 14. The immunoreactive avian ß-defensin 2 and cathelicidin 1 proteins were localized in the leukocyte-like cells in the lamina propria, and they were aggregated in the form of small islands. We conclude that the expression of innate immune molecules, including TLR, inflammation-related cytokines, and antimicrobial peptides, in the cecum are altered in association with changes in the density or composition of the luminal microbiota during the early phase of life in chicks.

Intestinal inflammation induced by dextran sodium sulphate causes liver inflammation and lipid metabolism disfunction in laying hens.

Gut inflammation caused by various factors including microbial infection leads to disorder of absorption of dietary nutrients and decrease in egg production in laying hens. We hypothesized that intestinal inflammation may affect egg production in laying hens through its impact on liver function. Dextran sodium sulphate (DSS) is known to induce intestinal inflammation in mammals, but whether it also induces inflammation in laying hens is not known. The goal of this study was to assess whether oral administration of DSS is a useful model of intestinal inflammation in laying hens and to characterize the effects of intestinal inflammation on egg production using this model. White Leghorn hens (350-day old) were administrated with or without 0.9 g of DSS/kg BW in drinking water for 5 D (n = 8, each). All laid eggs were collected, and their whole and eggshell weights were recorded. Blood was collected every day and used for biochemical analysis. Liver and intestinal tissues (duodenum, jejunum, ileum, cecum, cecal-tonsil, and colon) were collected 1 D after the final treatment. These tissue samples were used for histological analysis and PCR analysis. Oral administration of DSS in laying hens caused 1) histological disintegration of the cecal mucosal epithelium and increased monocyte/macrophage infiltration and IL-1ß, IL-6, CXCLi2, IL-10, and TGFß-4 gene expression; 2) decreased egg production; 3) increased leukocyte infiltration and IL-1ß, CXCLi2, and IL-10 expression in association with a high frequency of lipopolysaccharide-positive cells in the liver; and 4) decreased expression of genes related to lipid synthesis, lipoprotein uptake, and yolk precursor production. These results suggested that oral administration of DSS is a useful method for inducing intestinal inflammation in laying hens, and intestinal inflammation may reduce egg production by disrupting egg yolk precursor production in association with liver inflammation.

Effects of the routine multiple vaccinations on the expression of innate immune molecules and induction of histone modification in ovarian cells of layer chicks.

The aim of this study was to determine whether vaccination affects the expression of Toll-like receptors (TLRs), cytokines, and avian ß-defensins (AvBDs) in the chick ovary with or without lipopolysaccharide (LPS) stimulation. White Leghorn female chicks were administered vaccines for infectious bronchitis, Marek's disease, Newcastle disease, and infectious bursal disease during the first 14 D after hatching and ovarian tissues were collected on day 21. Control chicks received water or dilution buffer in place of vaccine. In Experiment 1, ovarian tissues were incubated with or without LPS, and the expression of innate immune molecules (TLRs, cytokines, and AvBDs) was examined by real-time PCR. In Experiment 2, the levels of histone modification in fresh ovarian tissues were examined by western blot analysis. The results of Experiment 1 showed that, in vaccinated chick ovaries, the expression of TLR1-1, 2-1, 2-2, and 21 was up-regulated, whereas that of TLR1-2, 4, and 7 was down-regulated under LPS stimulation. Among the examined 6 cytokines, only the expression of TNFSF15 was lower in the ovaries of vaccinated chicks than that in control with or without LPS stimulation. The expression of AvBD1, 2, 4, and 7 was lower in the ovaries of vaccinated chicks than in control without LPS stimulation, and that of AvBD1 and 2 was also lower even in ovaries incubated with LPS. In Experiment 2, the density of di-methyl histone H3 (Lys9) and acetyl histone H3 (Lys9) was significantly higher in the vaccine group than in the control, whereas di-methyl and tri-methyl histone H3 (Lys4) and acetyl histone H3 (Lys27) did not show differences between the groups. These results suggest that vaccination positively or negatively affects the expression of innate immune molecules in the chick ovary including TLRs, TNFSF15, and AvBDs, and it may be associated with epigenetic reprogramming by histone modifications in ovarian cells. Thus, in the future, it may be possible to develop or improve vaccination programs for the enhancement of the innate immune system in the hen ovary.

Age-related modulation of the isthmic and uterine mucosal innate immune defense system in laying hens.

Sustained production of good quality eggs for longer production cycles is a challenge for poultry farms. The impact of aging on the mucosal immune defense in the isthmus and uterus of hens, where the eggshell membrane and eggshell are formed, remains obscure. Thus, the aim of this study was to determine whether aging affects the mucosal tight junction (TJ) proteins, the synthesis of antimicrobial peptides including avian ß-defensins (AvBDs) and cathelicidins (CATHs), and Toll-like receptors (TLRs) in the isthmus and uterus of laying hens. Young and aged White Leghorn laying hens (35 and 130 wk old, respectively) were used. Total RNA and protein contents were isolated from the isthmic and uterine mucosae of these hens to examine the expression of TJ proteins, AvBD, and CATH genes and AvBD proteins by the real-time polymerase chain reaction and western blotting. The results showed that the mRNA expression of TJ proteins, namely zonula occludin 2 in the isthmus and occludin in the uterus, was higher in aged hens than in young hens. Expression of 2 AvBD genes in the isthmus and 4 AvBD genes in the uterus was higher in aged hens than in young hens. However, the expression of AvBD proteins 1 and 11 was not altered by aging. Expressions of CATH genes were not affected by aging in the isthmus or uterus. Expression of TLR genes was higher in aged hens than in young hens in the isthmus, while their expression in the uterus was not affected by aging. It can be concluded that aged hens have a higher potential ability to express TJ proteins and AvBDs for mucosal defense in the isthmic and uterine mucosae than in young hens.

Short communication: Production of antimicrobial peptide S100A8 in the goat mammary gland and effect of intramammary infusion of lipopolysaccharide on S100A8 concentration in milk.

This study aimed to determine the production site of antimicrobial peptide S100A8 in the goat mammary gland and changes in its concentration in milk after lipopolysaccharide (LPS) challenge. Sixteen Tokara goats were used in this study for mammary gland tissue, blood leukocyte, and milk somatic cell collection and LPS challenge. The mRNA expression and protein localization of S100A8 in the mammary gland parenchyma and teat, blood leukocytes, and milk somatic cells were examined by reverse-transcription PCR and immunohistochemistry. The S100A8 concentration in milk was measured at 0 to 144 h after intramammary challenge of LPS by enzyme immunoassay. The mRNA of S100A8 was expressed in the parenchyma and teat, leukocytes isolated from blood, and milk somatic cells. Antimicrobial peptide S100A8 was immunolocalized in the outermost layer of the teat skin of udders with and without LPS infusion, whereas in the mammary gland it was immunolocalized only in the leukocytes infiltrated in the alveoli after LPS infusion. Antimicrobial peptide S100A8 was also immunolocalized in the blood and milk leukocytes. The number of S100A8-positive cells in milk was higher than that in blood. The concentration of S100A8 in milk increased significantly at 72 h after intramammary infusion of LPS. These results suggest that S100A8 is produced in the leukocytes and that its secretion into milk is affected by LPS stimulation.

Expression of pro- and anti-inflammatory cytokines and chemokines during the ovulatory cycle and effects of aging on their expression in the uterine mucosa of laying hens.

The aim of this study was to examine whether cytokines and chemokines expressed in the uterine mucosa play a role in the process of eggshell formation in the chicken uterus. Changes in the expression levels of pro- and anti-inflammatory cytokines and chemokines in the uterine mucosa during an ovulatory cycle (experiment 1) and effects of aging on their expression (experiment 2) were examined. In experiment 1, the expression of the pro-inflammatory cytokines IL1ß, IL6, TNFSF15, and IFNγ, and a chemokine CX3CL1 was found to increase during eggshell biomineralization (16 h following oviposition), while anti-inflammatory TGFß2 expression was found to increase at 4 h following oviposition. In experiment 2, a higher expression of the anti-inflammatory cytokines TGFß2 and TGFß3, and chemokines CXCLi2 and CX3CL1, was observed in aged hens than in young hens. A significantly higher number of macrophages and CD8+ T cells were observed in the uterine tissue of aged hens than in young hens. Furthermore, the expression of adhesion molecules associated with leukocytic infiltration was found to be higher in aged hens than in young hens. We conclude that the eggshell formation process may be affected by the pro- and anti-inflammatory cytokines and chemokines. The balanced expressions of these molecules might be disrupted in aged hens.

Enriching gluten-free rice pasta with soybean and sweet potato flours.

The development of innovative rice products is a way to exploiting and adding value to low-grade African rice varieties. To this purpose, rice-based pasta was enriched with flours from soybean and orange-fleshed sweet potato, that are common ingredients in the African tradition. Four different formulations based on pre-gelatinized rice flour and liquid egg albumen, and containing soybean and/or sweet potato (up to 20%) were prepared and characterized via a multidisciplinary approach. Soybean and sweet potato enrichment leads to a decrease in the pasta consistency and in significant changes in the color of the resulting samples, likely due to Maillard-type reactions. E-sensing approaches indicated that the sensory profile of the various pasta products strongly depends on the type of enrichment. Data collected after cooking suggest that both soybean and sweet potato have a role in defining the firmness and water absorption, as well as the optimum cooking time. Structural characterization of proteins in the uncooked products indicates the presence of protein aggregates stabilized by hydrophobic interactions and disulfide bonds in all samples, although structural properties of the aggregates related to specific compositional traits.

Changes in plasma concentrations of S100A7 and S100A8 in dairy cows during pregnancy.

This study was carried out to examine the changes in plasma concentrations of the Ca-binding antimicrobial proteins S100A7 and S100A8 during pregnancy in dairy cows. Holstein Friesian cows (n = 19) were inseminated with Holstein Friesian semen. Blood was collected at days 30, 60, 90, 120, 150, 180, 210, 240 and 270 after insemination. Plasma was used for measuring the concentrations of S100A7 and S100A8. Both S100A7 and S100A8 concentrations showed similar patterns during gestation; they increased during the midgestation, between days 90 and 180, and then declined before calving. The findings indicated that plasma concentrations of S100A7 and S100A8 did not change significantly during pregnancy in cows. Further studies are required to determine the roles of S100A7 and S100A8 in physiological function during pregnancy in dairy cows.

Innate antiviral immune response against infectious bronchitis virus and involvement of prostaglandin E2 in the uterine mucosa of laying hens.

Infectious bronchitis virus (IBV) is an enveloped RNA virus that causes deformities in eggshells. The aim of this study was to investigate the innate immune response to IBV, and to determine whether prostaglandin (PG) E2, which is synthesized during inflammation, is involved in the innate immune response in the uterine mucosa. The effects of intra-oviductal inoculation with attenuated IBV (aIBV) on the expression of viral RNA recognition receptors and innate antiviral factors were examined by real-time PCR and immunohistochemistry, and on PGE2 levels by ELISA. Then, the effects of PGE2 on the expression of innate antiviral factors in cultured uterine mucosal cells were examined. The results showed that the expression of RNA virus pattern recognition receptors (TLR3, 7, and MDA5), antimicrobial peptides (avian ß-defensins, including AvBD1, 2, 4-6 and cathelicidins, including CATH1 and 3), and interferons (IFNα, ß, γ, λ) were upregulated, and the expression of cyclooxygenase 2 (PG synthase) and the level of PGE2 were increased in the uterine mucosa following aIBV inoculation. The number of AvBD2-positive cells in the mucosa also increased in response to aIBV. In cultured mucosal cells (mainly epithelial), the expression of AvBD4, 10-13 and IFNα, ß, and λ was upregulated following incubation with 500 nM PGE2. These results suggest that the expression of viral RNA-recognition receptors, AvBDs, CATHs, and IFNs and PGE2 are induced by the IBV antigen, and that the expression of a different set of AvBDs is also induced by PGE2 in the cultured uterine mucosal cells. These antiviral factors may play a role in the protection of the uterine mucosa from IBV infection.

Soybean-Enriched Snacks Based on African Rice.

Snacks were produced by extruding blends of partially-defatted soybean flour with flours from milled or parboiled African-grown rice. The interplay between composition and processing in producing snacks with a satisfactory sensory profile was addressed by e-sensing, and by molecular and rheological approaches. Soybean proteins play a main role in defining the properties of the protein network in the products. At the same content in soybean flour, use of parboiled rice flour increases the snack's hardness. Electronic nose and electronic tongue discriminated samples containing a higher amount of soybean flour from those with a lower soybean flour content.

Monocyte CD64 expression as a novel biomarker for the disease activity of systemic lupus erythematosus.

OBJECTIVE: Interferon alpha (IFN-α) is a key cytokine associated with systemic lupus erythematosus (SLE). IFN-α induces the expression of CD64 on monocytes (mCD64). Although enhanced mCD64 expression has been reported in patients with SLE, it has never been assessed quantitatively. The aim of this study was to investigate whether or not mCD64 expression correlates with SLE disease activity. METHODS: The mCD64 expression levels were assessed quantitatively in 40 patients with active or inactive SLE by using flow cytometry. The mCD64 expression levels were subsequently compared with the SLE disease activity index (SLEDAI) and levels of existing SLE activity biomarkers, such as anti-DNA antibody, complements, and so on. RESULTS: The mCD64 expression was significantly higher in active disease than in inactive disease SLE (median molecules/cell, interquartile range: 34,648, 8174-24,932 and 20,865, 6357-21,503, respectively; p < 0.001). The levels of mCD64 expression strongly correlated with SLEDAI (r = 0.68, p < 0.001). CONCLUSION: The mCD64 expression is a simple and useful biomarker for evaluating disease activity in patients with SLE.

Expression of Toll-like receptors and effects of lipopolysaccharide on the expression of proinflammatory cytokines and chemokine in the testis and epididymis of roosters.

The aim of this study was to determine the expression profiles of Toll-like receptors (TLR) in the testis and epididymis of rooster and whether the expression of IL-1ß, IL-6, CXCLi2, and TLR-4 was affected by lipopolysaccharide (LPS), a TLR-4 ligand. Roosters were intravenously injected with LPS or phosphate-buffered saline. Testes and epididymis were collected before and after 3 or 6 h postinjection. Total RNA was isolated from those tissues and expression of TLR and proinflammatory cytokines was analyzed by reverse-transcription PCR and quantitative real-time PCR. Reverse-transcription PCR analysis revealed that 7 of the known 10 chicken TLR in the testis and 9 of 10 in the epididymis were expressed. Expression of TLR-4 was found in both tissues. Expression of TLR-4 was significantly upregulated by LPS in the testis but not in the epididymis. Injection with LPS upregulated the expression of IL-1ß, IL-6, and CXCLi2 in the testis and epididymis by 3 to 6 h postinjection. However, injection with phosphate-buffered saline (control) did not affect their expression. These results suggest that proinflammatory cytokines and chemokine expression was upregulated by LPS probably through TLR-4 activation, and thus the reproductive tissues are comprehensively equipped to deal with a pathogenic insult.

Effects of lipopolysaccharide on the expression of proinflammatory cytokines and chemokines and the subsequent recruitment of immunocompetent cells in the oviduct of laying and molting hens.

The goal of this study was to examine whether lipopolysaccharide (LPS) induces the expression of proinflammatory cytokines and chemokines and recruits T cells in the lower part of the oviduct, and whether that response to LPS is different between the laying and molting phase. White Leghorn laying and molting hens were intravenously injected with saline (control) or LPS. The uterus and vagina of oviducts were collected 3 or 6 h after injection, and used for reverse transcription PCR analysis of IL-1ß, IL-6, IL-8 (CXCLi2), and lymphotactin (Lptn), and for immunohistochemical analysis for the frequency of CD4+ and CD8+ T cells. The expressions of IL-1ß, IL-6, and CXCLi2 in the uterus and that of IL-1ß in the vagina were upregulated in response to LPS 3 or 6 h after injection in both laying and molting hens. The CXCLi2 expression in the vagina was upregulated by LPS in laying hens, whereas those effects of LPS were not significant in molting hens. Expression of Lptn showed a tendency to be downregulated after 3 h, with recovery by 6 h after LPS injection. The frequency of CD4+ T cells tended to increase in response to LPS after 6 h in the lamina propria of the uterus and vagina in both laying and molting hens. The CD8+ T cell frequencies in the lamina propria of the uterus and vagina of laying hens increased in response to LPS after 6 h. However, in the molting hens, LPS stimulation resulted in CD8+ T cell increase in the vagina only and not in the uterus. These results suggest that expressions of proinflammatory cytokines and CXCLi2 chemokine are upregulated in association with T cell recruitment in response to LPS in the lower part of the oviduct, although CD8+ T cells in the uterus may be depressed during the molting phase. These immunoresponses may play roles in the defense against infection of the oviduct.

Molecular events involved in up-regulating human Na+-independent neutral amino acid transporter LAT1 during T-cell activation.

We investigated the regulation of system-L amino acid transporter (LAT1) during T-cell activation. In quiescent T-cells, L-leucine transport is mediated mainly by the system-L amino acid transport system and is increased significantly during T-cell activation by PMA and ionomycin. In quiescent T-cells, the LAT1 protein was heterocomplexed with 4F2 heavy chain (4F2hc) in the plasma membrane. During T-cell activation, the amounts of 4F2hc and LAT1 heterocomplex were significantly elevated compared with those in quiescent T-cells. In addition, by Northern-blot analysis, these increments were found to be due to elevated levels of LAT1 and 4F2hc mRNA. Transient expression of constructs comprising various LAT1 gene promoter fragments, which contained all three of the GC boxes, was sufficient for promoting luciferase expression in Jurkat T-cells, but the promoter of the LAT1 gene did not respond to PMA and ionomycin. Similar observations were observed in the human 4F2hc gene promoter. In nuclear run-on assay, the LAT1 and 4F2hc genes were actively transcribed even in quiescent T-cells, but the low levels of both transcripts were shown to be the result of a block to transcription elongation within the exon 1 intron 1 regions. These findings indicated that a removal of the block to mRNA elongation stimulates the induction of system-L amino acid transporter gene transcripts (LAT1 and 4F2hc) in activated T-cells.