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1.
J Dairy Sci ; 101(4): 3261-3270, 2018 Apr.
Article in English | MEDLINE | ID: mdl-29428750

ABSTRACT

Drying cows off at the end of lactation is a routine management practice in dairy operations. Most dairies in the United States and many other countries dry cows off abruptly (e.g., stop milking cows on a set day), which has been shown to affect cow comfort. Gradually reducing milk production is another approach to dry cows off, routinely used in some countries and herds. The objective of the study was to evaluate the effects of abrupt and gradual milk cessation and milk yield at the time on cow activity after dry-off. Daily lying time, number of lying bouts per day, average lying bout length, and steps taken per day by abruptly and gradually dried-off cows were monitored by data loggers for 2 wk before and after the final milking at the end of lactation. Gradual cows were milked once daily for the last week of lactation, and abrupt cows were milked as usual (3×/d) until the end of lactation. Gradual cessation of milking significantly reduced milk yield by the day of dry-off. After dry-off, gradual cows tended to have longer lying bouts than abrupt cows, but no other differences in cow activity between the 2 treatments were observed. Regardless of the dry-off method, the average length of a lying bout decreased by 4 min and total daily lying time decreased by 19 min after dry-off for each 5-kg increase in milk yield before dry-off. Lying behavior of primiparous cows was more affected by the level of milk yield at dry-off than that of older cows. A reduction in lying times with increasing milk yield may indicate discomfort due to the accumulating milk in the udder. Using a method that lowers milk production before dry-off and managing primiparous and multiparous cows separately around dry-off are beneficial for cow comfort after dry-off.


Subject(s)
Behavior, Animal , Cattle/physiology , Dairying/methods , Lactation/physiology , Milk/metabolism , Animals , Databases, Genetic , Environment , Female , Mammary Glands, Animal/physiology , Parity
2.
J Dairy Sci ; 100(3): 2080-2089, 2017 Mar.
Article in English | MEDLINE | ID: mdl-27988116

ABSTRACT

The objective of this study was to assess the effect of milk cessation method (abrupt or gradual) at dry off on milk yield and somatic cell score (SCS) up to 120 d in milk during the subsequent lactation. Data from 428 cows from 8 dairy herds in Ohio were analyzed. Abrupt cessation cows kept the farm's regular milking schedule (2 or 3 times) through dry off and gradual cessation cows were milked once daily for the final week of lactation. Milk yield and SCS were collected using Dairy Herd Improvement Association test-day records. Aseptic quarter milk samples were collected approximately 1 wk before dry off, at dry off, and within 1 wk after calving for bacterial culture to determine the presence of intramammary infections. Overall, milk cessation method was not significantly associated with either milk yield or SCS in early lactation; however, interaction between the milk cessation method and herd was highly significant. Cows producing greater amounts of milk around dry off had significantly higher SCS in the following lactation. Shorter dry periods were significantly associated with decreased milk yield in the following lactation, especially among abruptly dried off cows. Additionally, as expected, several other factors, such as parity of cows and stage of lactation, were significantly associated with both outcomes. No interactions between the milk cessation method and the other explanatory variables in the final models were significant. The results of the current study suggest that higher milk yield at dry off was associated with higher SCS in the following lactation, even though milk cessation method at the end of lactation had a varying effect on test-day milk yield and SCS in different herds during the first 120 d in milk in the following lactation. The specific herd characteristics influencing this could not be identified within this study, warranting further research.


Subject(s)
Lactation , Milk/microbiology , Animals , Cattle , Dairying , Female , Ohio , Parity , Time Factors
3.
J Dairy Sci ; 99(5): 4005-4017, 2016 May.
Article in English | MEDLINE | ID: mdl-26898287

ABSTRACT

The objective of this study was to evaluate the effect of milking cessation method (abrupt or gradual) and daily milk yield before dry-off on milk leakage following dry-off and intramammary infections (IMI) at calving. Data from 1,086 quarters of 285 cows from 5 Ohio dairy herds were analyzed. All cows that were due to be dried off within a week were assigned to the same study group to facilitate management. Abrupt-cessation cows kept the farm's regular milking schedule through dry-off, and gradual-cessation cows were milked once daily for the final week of lactation. Aseptic technique was used to collect quarter foremilk samples at the time of enrollment (7 to 14 d before expected dry-off), the final milking before dry-off (D-O), and within 7 d of calving. Cows in the gradual-cessation group were observed for milk leakage during the period of once-daily milking. In the only herd that did not use internal teat sealants at dry-off, milk leakage after dry-off was recorded in both abrupt and gradual groups. Gradual cessation decreased milk production by 33.4% during the final week of lactation, causing milk yield at D-O to be lower for these cows compared with abrupt-cessation cows (13.2 vs. 19.8kg/d, respectively). Logistic regression models were used to model the probability of a quarter being infected at calving with any pathogen, accounting for clustering of quarters within cows and cows within herds. The final model investigating the probability of IMI at calving was stratified by parity of cows at the time of dry-off (primiparous and multiparous). Among quarters of cows that ended their first lactation, abrupt cessation of milking before dry-off and milk leakage after dry-off were associated with an increased risk of IMI at calving. Among quarters of multiparous cows, on the other hand, gradual cessation of milking before dry-off, presence of IMI at D-O, and thrice-daily milking during lactation increased the odds of IMI at calving. These results indicate that implementation of differing management practices near dry-off for different parity groups may improve mammary health within a herd.


Subject(s)
Cattle Diseases/etiology , Dairying/methods , Mammary Glands, Animal/physiopathology , Milk/metabolism , Animals , Cattle , Cattle Diseases/physiopathology , Female , Lactation , Logistic Models , Ohio , Parity
4.
J Dairy Sci ; 98(3): 1786-96, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25547311

ABSTRACT

Endotoxin tolerance (ET) can develop in mammals that have been challenged repeatedly with sublethal amounts of lipopolysaccharide (LPS). Previous research has shown that subclinical ruminal acidosis can increase circulating concentrations of LPS. We investigated whether ET would develop in Holstein cows that were subjected to chronic subacute ruminal acidosis (SARA) or acute SARA followed by intramammary infusion of LPS. Twenty-four cows, both primiparous and multiparous, were assigned to 8 blocks of 3 cows. Cows within blocks were randomly assigned to 1 of 3 treatments: (1) control (diet DM was 24% starch and 35% NDF), (2) high starch (formulated to induce chronic milk fat depression with 29% starch and 32% NDF), and (3) acidosis (designed to cause acute bouts of milk fat depression by short-term feeding of a diet with 32% starch, some of which came from wheat grain, and 30% NDF). Cows on the control and high-starch treatments were fed their respective diets throughout the 24-d trial. The acidosis cows were fed the control diet during most of the experiment, except during two 2-d bouts (d 10 and 11 and 17 and 18 of the experiment) in which a high-starch diet was fed. Cows on the high-starch and acidosis treatments produced milk fat with an altered fatty acid profile indicative of SARA (e.g., increased concentrations of specific trans, and odd-, and branched-chain fatty acids), but only cows on the high-starch treatment had milk fat depression. Concentrations of serum amyloid A were elevated in cows on the acidosis treatment, but did not differ between control and high-starch cows. On d 20 of the experiment, all cows were given an intramammary infusion of 10 µg of LPS into 1 mammary quarter 3h after morning milking. Milk yield and DMI decreased the day of the infusion, but the response was not affected by dietary treatment. No systemic indicators of ET were observed among treatments, but evidence of an ET response at the local level of the mammary gland was observed. Cows fed the control diet had higher concentrations of serum amyloid A in milk 12 and 24h postinfusion than did cows fed the high-starch diet and higher concentrations than cows on the acidosis treatment at 12h postinfusion. Our data suggest cows that experienced varying degrees of SARA (based on altered milk fatty acid profile) and subsequent experimental endotoxin mastitis experienced a blunted inflammatory response at the level of the mammary gland, but not a systemic reduction in some inflammatory mediators.


Subject(s)
Acidosis/veterinary , Animal Feed , Cattle Diseases/etiology , Immunologic Deficiency Syndromes/veterinary , Lipopolysaccharides/administration & dosage , Starch/administration & dosage , Acidosis/etiology , Animals , Cattle , Diet/veterinary , Dietary Carbohydrates/administration & dosage , Fatty Acids , Female , Lactation , Milk/drug effects , Parity , Random Allocation , Toll-Like Receptor 4
5.
J Dairy Sci ; 94(6): 3045-53, 2011 Jun.
Article in English | MEDLINE | ID: mdl-21605774

ABSTRACT

Interactions of sources and processing methods for nonstructural carbohydrates may affect the efficiency of animal production. Five rumen-cannulated cows in late lactation were placed in a 5 × 5 Latin square design and fed experimental diets for 2 wk. In the production trial, 54 cows were fed the experimental diets for 12 wk beginning at d 60 in milk. Diets contained 24% corn silage and 22% hay, averaging 20% alfalfa and 2% grass but being adjusted as needed to maintain dietary concentrations of 36% neutral detergent fiber. The control diet contained steam-flaked corn (SFC) and the other diets contained either finely (FGC; 0.8 mm) or coarsely ground corn (CGC; 1.9 mm), factorialized with or without 3.5% liquid feed (LF). The LF diets provided 1.03% of dietary dry matter as supplemental sugar. The FGC decreased rumen pH and concentration of NH(3)N compared with CGC. The SFC and FGC tended to increase the molar percentage of ruminal propionate and decrease the acetate:propionate ratio. The LF increased molar percentage of ruminal butyrate with FGC but not CGC. The LF tended to decrease starch digestibility with the CGC but not with the FGC. As expected, the SFC and FGC increased total tract starch digestibility. The DMI and milk yield were similar among dietary treatments. Compared with ground corn diets, the SFC tended to decrease milk fat percentage; thus, 3.5% fat-corrected milk and feed efficiency were decreased with SFC. The LF decreased milk protein percentage but had no effect on milk protein yield. The SFC compared with dry ground corn decreased the concentration of milk urea nitrogen. Sugar supplementation using LF appeared to be more beneficial with FGC than CGC. Increasing the surface area by finely grinding corn is important for starch digestibility and optimal utilization of nutrients.


Subject(s)
Cattle/physiology , Diet/veterinary , Dietary Sucrose/administration & dosage , Digestion/physiology , Lactation/physiology , Zea mays/metabolism , Animal Feed , Animal Nutritional Physiological Phenomena , Animals , Cattle/metabolism , Female , Milk/chemistry , Milk/metabolism , Rumen/metabolism , Zea mays/chemistry
6.
Gut ; 52(9): 1297-303, 2003 Sep.
Article in English | MEDLINE | ID: mdl-12912861

ABSTRACT

BACKGROUND: and aims: Tumour necrosis factor alpha (TNF-alpha) induction of nuclear factor kappaB (NFkappaB) activation plays a major role in the pathogenesis of inflammatory bowel disease (IBD). Trefoil factor family peptides TFF1, TFF2, and TFF3 exert protective, curative, and tumour suppressive functions in the gastrointestinal tract. In this study, we investigated effects of the TNF-alpha/NFkappaB regulatory pathway by TNF-alpha on expression of TFFs. METHODS: After TNF-alpha stimulation, expression of TFF genes was analysed by quantitative real time polymerase chain reaction and by reporter gene assays in the gastrointestinal tumour cell lines HT-29 and KATO III. Additionally, NFkappaB subunits and a constitutive repressive form of inhibitory factor kappaB (IkappaB) were transiently coexpressed. In vivo, morphological changes and expression of TFF3, mucins, and NFkappaB were monitored by immunohistochemistry in a rat model of 2,4,6-trinitrobenzene sulphonic acid induced colitis. RESULTS: TNF-alpha stimulation evoked up to 10-fold reduction of TFF3 expression in the colon tumour cell line HT-29. Downregulation of reporter gene transcription of TFF3 was observed with both TNF-alpha and NFkappaB, and was reversible by IkappaB. In vivo, the increase in epithelial expression of NFkappaB coincided with reduced TFF3 expression during the acute phase of experimental colitis. CONCLUSIONS: Downregulation of intestinal trefoil factor TFF3 is caused by repression of transcription through TNF-alpha and NFkappaB activation in vitro. In IBD, perpetual activation of NFkappaB activity may contribute to ulceration and decreased wound healing through reduced TFF3.


Subject(s)
NF-kappa B/physiology , Neuropeptides/metabolism , Tumor Necrosis Factor-alpha/physiology , Animals , Colitis/chemically induced , Colitis/metabolism , Down-Regulation , Gene Expression Regulation , Genes, Reporter , HT29 Cells , Humans , Luciferases/genetics , Luciferases/metabolism , Models, Animal , NF-kappa B/antagonists & inhibitors , Neuropeptides/genetics , Polymerase Chain Reaction , Rats , Trefoil Factor-2 , Trefoil Factor-3 , Tumor Cells, Cultured
7.
Gut ; 51(5): 685-90, 2002 Nov.
Article in English | MEDLINE | ID: mdl-12377807

ABSTRACT

BACKGROUND: Damage to the gastrointestinal mucosa results in the acute up-regulation of the trefoil factor family peptides TFF1, TFF2, and TFF3. They possess protective, healing, and tumour suppressive functions. Little is known about the regulation of TFF gene expression. The promoters of all three TFF genes contain binding sites (E box) for upstream stimulating factor (USF) and Myc/Max/Mad network proteins. AIMS: To determine the nature and function of transcription factors that bind to these E boxes and to understand their role for TFF gene expression. METHODS: TFF promoter activities were determined by reporter gene assays. DNA binding was monitored by electromobility shift assays and by chromatin immunoprecipitation analyses. Expression of endogenous TFF was determined by multiplex RT-PCR. RESULTS: It was observed that the TFF2 promoter is specifically and efficiently activated by USF transcription factors but not by c-Myc. USF displayed comparable binding to a high affinity Myc/Max binding site compared with the three TFF E boxes, while c-Myc exhibited lower affinity to the TFF E boxes. In contrast, pronounced binding differences were observed in cells with a strong preference for USF to interact specifically with the TFF2 E box, while Myc was not above background. Exogenous expression of USF was sufficient to activate the chromosomal TFF2 and to a lesser extent, the TFF1 gene. CONCLUSION: These findings define USF factors as regulators of the TFF2 gene and suggest that promoter specific effects are important for a pronounced gene activation of this cytoprotective peptide.


Subject(s)
DNA-Binding Proteins , Gastrointestinal Neoplasms/genetics , Gene Expression Regulation , Growth Substances/genetics , Mucins , Muscle Proteins , Neuropeptides , Peptides/genetics , Promoter Regions, Genetic , Proto-Oncogene Proteins c-myc/genetics , Transcription Factors/genetics , Binding Sites , Electrophoretic Mobility Shift Assay , Gastrointestinal Neoplasms/metabolism , Growth Substances/metabolism , Humans , Peptides/metabolism , Precipitin Tests , Proto-Oncogene Proteins c-myc/metabolism , Reverse Transcriptase Polymerase Chain Reaction , Transcriptional Activation , Trefoil Factor-2 , Trefoil Factor-3 , Tumor Cells, Cultured , Upstream Stimulatory Factors
8.
Viral Immunol ; 15(1): 177-92, 2002.
Article in English | MEDLINE | ID: mdl-11952140

ABSTRACT

Hantavirus nucleocapsid protein (N) has been proven to induce highly protective immune responses in animal models. The knowledge on the mechanisms behind N-induced protection is still limited, although recent data suggest that both cellular and humoral immune responses are of importance. For a detailed B-cell epitope mapping of Puumala hantavirus (PUUV) N, we used recombinant N derivatives of the Russian strain CG18-20 and the Swedish strain Vranica/Hällnäs, as well as overlapping synthetic peptides corresponding to the Finnish prototype strain Sotkamo. The majority of a panel of monoclonal antibodies (mAbs) reacted with proteins derived from all included PUUV strains demonstrating the antigenic similarity of these proteins. In line with previous results, the epitopes of most mAbs were mapped within the 80 N-terminal amino acids of N. The present study further revealed that the epitopes of four mAbs raised against native viral N were located within amino acids 14-45, whereas one mAb raised against recombinant N was mapped to amino acids 14-39. Differences between the reactivity of the PUUV strains Vranica/Hällnäs and CG18-20 N suggested the importance of amino acid position 35 for the integrity of the epitopes. In line with the patterns obtained by the truncated recombinant proteins, mapping by overlapping peptides (PEPSCAN) confirmed a complex recognition pattern for most analyzed mAbs. Together, the results revealed the existence of several, partially overlapping, and discontinuous B-cell epitopes. In addition, based on differences within the same competition group, novel epitopes were defined.


Subject(s)
Epitope Mapping/methods , Epitopes, B-Lymphocyte , Nucleocapsid/immunology , Puumala virus/immunology , Amino Acid Sequence , Animals , Antibodies, Monoclonal/immunology , Mice , Molecular Sequence Data , Nucleocapsid/chemistry
9.
FEBS Lett ; 488(3): 206-10, 2001 Jan 19.
Article in English | MEDLINE | ID: mdl-11163773

ABSTRACT

Trefoil factor family (TFF) peptides promote cell migration, heal the mucosa and may suppress tumor growth. In reporter gene assays we show that aspirin (1-12 mM) evokes a six-fold up-regulation of TFF2, but not TFF1 and TFF3 transcription in human gastrointestinal cell lines. 6 h after application up-regulation of endogenous TFF2 mRNA was observed. TFF2 transcription was enhanced by indomethacin and arachidonic acid but repressed by staurosporine, suggesting mediation via protein kinase C. We mapped an aspirin responding element -546 to -758 bp upstream of TFF2. Up-regulation of TFF2 by aspirin may partially explain the chemopreventive potential of low dose aspirin in gastrointestinal carcinogenesis.


Subject(s)
Aspirin/pharmacology , Gastrointestinal Neoplasms/genetics , Growth Substances/genetics , Mucins , Muscle Proteins , Neuropeptides , Peptides/genetics , Transcriptional Activation/drug effects , Arachidonic Acid/metabolism , Arachidonic Acid/pharmacology , Gastrointestinal Neoplasms/metabolism , Genes, Reporter/genetics , Humans , Indomethacin/pharmacology , Models, Biological , Protein Kinase C/antagonists & inhibitors , RNA, Messenger/genetics , RNA, Messenger/metabolism , Response Elements/genetics , Sequence Deletion/genetics , Signal Transduction/drug effects , Staurosporine/pharmacology , Transfection , Trefoil Factor-2 , Trefoil Factor-3 , Tumor Cells, Cultured
10.
Biochim Biophys Acta ; 1490(3): 324-32, 2000 Feb 29.
Article in English | MEDLINE | ID: mdl-10684977

ABSTRACT

One of the early events in inflammation and epithelial restitution of the gastrointestinal tract is the up-regulation of secretory peptides belonging to the trefoil factor family (TFF) that promote cell migration, protect and heal the mucosa. Their major expression site is stomach (TFF1, TFF2) and intestine (TFF3). Located in the 5'-flanking region of the genes are several consensus sites for members of the GATA transcription factors known to control gut-specific gene expression. By reverse transcription-PCR (RT-PCR), GATA-6 was shown to be expressed in a variety of tumor cell lines of gastric, intestinal and pancreatic origin. In MKN45, KATOIII and LS174T, cotransfection with TFF reporter genes and GATA-6 expression vectors revealed that GATA-6 activates TFF1 and TFF2 4-6-fold, without an effect on TFF3. The functional contribution of GATA binding sequences in the reverse orientation was further characterized by reporter gene assays using TFF2 deletion constructs and by gel shift experiments.


Subject(s)
DNA-Binding Proteins/metabolism , Growth Substances/genetics , Mucins , Muscle Proteins , Neuropeptides , Peptides/genetics , Proteins/genetics , Transcription Factors/metabolism , Transcriptional Activation , Adenocarcinoma , Binding Sites , GATA6 Transcription Factor , Genes, Reporter , Humans , Promoter Regions, Genetic , RNA, Messenger/metabolism , Stomach Neoplasms , Trefoil Factor-1 , Trefoil Factor-2 , Trefoil Factor-3 , Tumor Cells, Cultured , Tumor Suppressor Proteins
11.
J Pathol ; 190(2): 133-42, 2000 Feb.
Article in English | MEDLINE | ID: mdl-10657010

ABSTRACT

Trefoil factor family domain peptides (TFF) are thought to be involved in mucosal epithelial restitution and wound healing of the gastrointestinal tract and are up-regulated in ulceration and in a variety of solid tumours. It was hypothesized that TFFs are also expressed on mucosal surfaces of the human respiratory tract. Lung tissue, nasal polyps, and sputum samples from seven patients with cystic fibrosis (CF), two with chronic and acute bronchitis, and non-dysplastic material from two cases of bronchial adenocarcinoma were analysed for TFF expression by immunohistochemistry, immunofluorescence, western blot and RT-PCR. Expression of TFF1 and TFF3 was observed in material from all patients. TFFs were localized in goblet and ciliated cells, as well as in some submucosal cells of tracheobronchial tissues and nasal polyps from normal and CF individuals. In sputa of patients with CF and with chronic or acute bronchitis, TFF1 and TFF3 were detected by western blotting. Freshly cultivated nasal epithelial cells transcribed and secreted TFFs and mucins, whereas nasal cells cultivated for 6 weeks still expressed mucins, but not TFFs. Secreted TFFs and mucins also bound to the surface of Staphylococcus aureus in infected CF airways. In conclusion, TFF1 and TFF3 are expressed and secreted in normal and inflamed airways. The association of TFFs with bacteria may contribute to the anti-microbial mucociliary defence system.


Subject(s)
Mucins , Muscle Proteins , Neuropeptides , Proteins/metabolism , Respiratory Tract Diseases/metabolism , Adenocarcinoma/metabolism , Aged , Bronchitis/metabolism , Chronic Disease , Cystic Fibrosis/metabolism , Female , Fluorescent Antibody Technique, Indirect , Growth Substances/metabolism , Humans , Immunoenzyme Techniques , Lung Neoplasms/metabolism , Male , Middle Aged , Nasal Polyps/metabolism , Peptides/metabolism , Sinusitis/metabolism , Sputum/metabolism , Staphylococcus aureus/metabolism , Trefoil Factor-1 , Trefoil Factor-2 , Trefoil Factor-3 , Tumor Suppressor Proteins
12.
Gene ; 236(1): 133-6, 1999 Aug 05.
Article in English | MEDLINE | ID: mdl-10433974

ABSTRACT

Mammalian trefoil factor family (TFF)-domain peptides promote gastrointestinal protection, healing and cell migration and may act as tumour suppressors. TFF-like domains also constitute modules of composite proteins like mucin glycoproteins and zona pellucida proteins. Database searches with a modified, less stringent consensus sequence - C-x(5,6)-[ST]-x(3)-C-x(4,5)-C-C-[FYWH]-x(2, 24)-C-[FY] - revealed that ancestors of the TFF-domain arose before amphibian evolution. Eggshell proteins and a zona pellucida-like protein in teleost species, an epidermis-specific protein in a tunicate as well as an open reading frame in a nematode exhibited TFF-like motifs suggesting that they most likely originated in some multicellular organism.


Subject(s)
Evolution, Molecular , Growth Substances/genetics , Mucins , Muscle Proteins , Neuropeptides , Peptides/genetics , Amino Acid Sequence , Animals , Chromosome Mapping , Chromosomes, Human, Pair 21 , Exons , Humans , Molecular Sequence Data , Sequence Homology, Amino Acid , Trefoil Factor-2 , Trefoil Factor-3 , Urochordata/genetics
13.
J Pathol ; 187(5): 541-8, 1999 Apr.
Article in English | MEDLINE | ID: mdl-10398119

ABSTRACT

The expression of two trefoil peptides (TFF1 and TFF2) and four mucins (MUC1, MUC2, MUC5AC, and MUC6) was evaluated by immunohistochemistry and reverse transcription-polymerase chain reaction (RT-PCR) in 29 gastric polyps, 10 hyperplastic and 19 adenomatous, eight of which displayed malignant transformation. The aims of this study were to characterize the expression profile of these molecules in each type of polyp and to investigate possible modifications of the profile during the process of malignant transformation. All hyperplastic polyps displayed immunoreactivity for TFF1, MUC5AC, and MUC1 in more than 75 per cent of the cells. In adenomatous polyps, three main phenotypes could be identified: complete gastric phenotype (co-expression of TFF1 and MUC5AC)-nine cases (47.4 per cent); incomplete gastric phenotype (TFF1-positive and MUC5AC-negative)-seven cases (36.8 per cent); non-gastric (intestinal) phenotype (no expression of TFF1 or MUC5AC)-three cases (15.8 per cent). Data yielded by immunohistochemistry and RT-PCR showed a good correlation for both TFF1 and TFF2. One hyperplastic and seven adenomatous polyps with villous architecture displayed foci of diffuse and intestinal-type carcinoma, respectively; in all of these cases, MUC1 expression and signs of gastric differentiation were observed in both the non-malignant and the carcinomatous component. It is concluded that gastric differentiation is a feature of hyperplastic polyps and of a subset of adenomatous polyps which is shared by early carcinomas arising in some of these polyps, regardless of the histological type of polyp and of carcinoma.


Subject(s)
Growth Substances/metabolism , Heat-Shock Proteins/metabolism , Mucins/metabolism , Muscle Proteins , Neoplasm Proteins/metabolism , Neuropeptides , Peptides/metabolism , Polyps/metabolism , Precancerous Conditions/metabolism , Proteins/metabolism , Stomach Neoplasms/metabolism , Adenomatous Polyps/metabolism , Cell Differentiation , Cell Transformation, Neoplastic , Humans , Hyperplasia/metabolism , Hyperplasia/pathology , Immunoenzyme Techniques , Polyps/pathology , Precancerous Conditions/pathology , Reverse Transcriptase Polymerase Chain Reaction , Stomach/pathology , Stomach Neoplasms/pathology , Trefoil Factor-1 , Trefoil Factor-2 , Trefoil Factor-3 , Tumor Suppressor Proteins
14.
DNA Cell Biol ; 18(2): 157-64, 1999 Feb.
Article in English | MEDLINE | ID: mdl-10073575

ABSTRACT

The winged helix transcription factors HNF-3/FKH (forkhead homologs) activate endodermal-derived and acute-phase gene expression and control gut development in Drosophila. Trefoil factor family (TFFs) peptides are vertebrate products secreted by mucin-producing epithelial cells of the gastrointestinal tract involved in restitution and repair of the mucosa. They are positively regulated in ulcerative and neoplastic conditions. We describe a consensus sequence in human and rodent TFF promoters close to the TATAA box showing striking similarity to the binding site of the HNF-3/FKH family. In gel retardation assays, HNF-3 alpha and beta bound predominantly to the site in TFF1 (formerly pS2) and, to a lesser extent, to the sites in TFF2 or TFF3. Mutations generated in this motif severely impaired transcription of TFF1 reporter genes. Cotransfection with expression vectors of HNF-3alpha and beta, but not the related HFH 11A and B, specifically activated the wild-type TFF1 reporter genes. Activation of endogenous expression of TFF1 by HNF-3 alpha and beta gene products was more than 1000 fold in the pancreatic cell line Capan-2 and fivefold in the gastric cell line MKN-45, whereas the intestinal cell lines HUTU 80 and HT-29 displayed no effect. Thus, HNF-3/FKH factors contribute causally to cell-specific regulation of TFF genes and may explain the acute-phase response of TFF peptides.


Subject(s)
Growth Substances/genetics , Mucins , Muscle Proteins , Neuropeptides , Peptides/genetics , Proteins/genetics , Trans-Activators/genetics , Animals , Base Sequence , Binding Sites , Cell Nucleus/chemistry , Cell Nucleus/metabolism , Cell-Free System/chemistry , Cell-Free System/metabolism , DNA-Binding Proteins/genetics , Forkhead Box Protein M1 , Forkhead Transcription Factors , Gene Expression Regulation, Neoplastic , Genes, Reporter/genetics , Growth Substances/metabolism , Hepatocyte Nuclear Factor 3-alpha , Hepatocyte Nuclear Factor 3-beta , Humans , Molecular Sequence Data , Mutagenesis , Nuclear Proteins/genetics , Peptides/metabolism , Promoter Regions, Genetic , Protein Binding , Proteins/metabolism , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Sequence Homology, Nucleic Acid , TATA Box , Trans-Activators/metabolism , Transcription Factors/genetics , Trefoil Factor-1 , Trefoil Factor-2 , Trefoil Factor-3 , Tumor Cells, Cultured/chemistry , Tumor Cells, Cultured/cytology , Tumor Cells, Cultured/metabolism , Tumor Suppressor Proteins
15.
Neurology ; 52(4): 746-9, 1999 Mar 10.
Article in English | MEDLINE | ID: mdl-10078721

ABSTRACT

OBJECTIVES: To determine the relative magnitudes of neuron-specific enolase (NSE) levels after complex partial status epilepticus (SE), absence SE, generalized convulsive SE, and subclinical generalized convulsive SE (frequently referred to as acute symptomatic myoclonic status epilepticus). BACKGROUND: NSE is a marker of acute brain injury and blood-brain barrier dysfunction, which is elevated in SE. METHODS: Serum NSE levels were drawn in 31 patients 1, 2, 3, and 7 days after SE. Patients were classified as acute symptomatic or remote symptomatic, and the duration and outcome of SE were determined and correlated with the peak NSE level. RESULTS: NSE was elevated significantly in all four subtypes of SE, but NSE levels were highest in complex partial and subclinical SE. The mean peak NSE level for the complex partial SE group was 23.88 ng/mL (n = 12), 21.5 ng/mL for absence SE (n = 1), 14.10 ng/mL for the generalized convulsive SE group (n = 12), and 37.83 ng/mL for the subclinical SE group (n = 6), all of which was significantly higher than normal control subjects (5.02 ng/mL). Outcome was significantly different between the three groups (p = 0.0007), and was significantly worse for subclinical SE (p = 0.0005, subclinical versus generalized convulsive SE). CONCLUSION: Serum NSE levels were highest in complex partial and subclinical generalized convulsive SE. The extremely high levels of NSE in subclinical SE reflect the severity of the acute neurologic insults and poor outcome common to subclinical SE. High NSE levels in complex partial SE reflects the long duration of SE in this subgroup, and potential for brain injury.


Subject(s)
Phosphopyruvate Hydratase/blood , Status Epilepticus/blood , Electroencephalography , Glasgow Coma Scale , Humans , Prognosis , Prospective Studies , Status Epilepticus/physiopathology , Time Factors
16.
Emerg Infect Dis ; 5(1): 168-71, 1999.
Article in English | MEDLINE | ID: mdl-10081688

ABSTRACT

An outbreak of infectious diarrhea with 70 laboratory-confirmed cases (58 with Giardia lamblia) and 107 probable cases occurred in U.K. tourists who stayed in a hotel in Greece. After a cluster of six cases in persons who had stayed at the hotel was reported, the Communicable Disease Surveillance Centre began active case ascertainment. This outbreak illustrates the value of an approach to surveillance that integrates routine surveillance data with active case ascertainment.


Subject(s)
Diarrhea/parasitology , Disease Outbreaks , Giardia lamblia/isolation & purification , Giardiasis/parasitology , Travel , Adolescent , Adult , Animals , Child , Diarrhea/epidemiology , Giardiasis/epidemiology , Greece/epidemiology , Humans , Male , Risk Factors
17.
Hum Hered ; 49(1): 45-7, 1999 Jan.
Article in English | MEDLINE | ID: mdl-9858857

ABSTRACT

Peptides belonging to the trefoil factor family (TFF) protect the gastrointestinal epithelia. Overexpression of TFFs was observed in pathological conditions such as gastritis, ulceration, metaplasia and neoplasia of the gastrointestinal tract. The aims of this work were to investigate the recently described TFF2 gene polymorphism in different European populations. DNA samples from blood of healthy individuals and gastric cancer patients were genotyped using the polymerase chain reaction. They were compared to a gastric cancer population. The results do not show any significant difference in allelic frequencies between gastric cancer patients and healthy individuals from Portugal. However, the frequency of the two alleles found varies considerably among Europeans.


Subject(s)
Gene Frequency , Growth Substances/genetics , Mucins , Muscle Proteins , Neuropeptides , Peptides/genetics , Polymorphism, Genetic , Stomach Neoplasms/genetics , Alleles , Europe , Genetic Predisposition to Disease , Humans , Polymerase Chain Reaction , Tandem Repeat Sequences , Trefoil Factor-2 , Trefoil Factor-3
19.
Int J Mol Med ; 2(3): 353-61, 1998 Sep.
Article in English | MEDLINE | ID: mdl-9855711

ABSTRACT

A group of secreted peptides (trefoil factor family; TFF) is abundantly expressed at mucosal surfaces of the gastrointestinal tract and promote epithelial restitution. They are upregulated around areas of epithelial damage, ulceration and neoplasia. The transcriptional regulation of the three human TFF genes was assayed by multiplex RT-PCR and reporter gene analysis in 8 gastrointestinal carcinoma cell lines. The level of endogenous mRNA matched well the reporter gene activity of all TFFs, indicating that the cis-acting elements located less than 1,000 bp upstream of the TATAA box account for cell-specific gene expression. In HT-29, the endogenous TFF expression profile changed in relation to cell growth conditions. Deletion and mutation analysis of TFF promoter constructs revealed enhancing elements shared within the three TFF promoters that were shown to bind nuclear proteins. Thus such specific DNA-protein interaction may explain the TFF peptides' cell specific expression pattern and altered levels in pathological conditions.


Subject(s)
5' Flanking Region , Peptides/metabolism , Cell Line, Tumor , Gene Expression Regulation , Humans , Intestinal Neoplasms , Nuclear Proteins/metabolism , Pancreatic Neoplasms , Peptides/genetics , Promoter Regions, Genetic , Protein Binding , Reverse Transcriptase Polymerase Chain Reaction , Stomach Neoplasms , Trefoil Factor-2 , Trefoil Factor-3
20.
FEBS Lett ; 439(1-2): 180-4, 1998 Nov 13.
Article in English | MEDLINE | ID: mdl-9849902

ABSTRACT

The gastrointestinal tract is exposed to environmental insult as a result of food intake or in pathological conditions such as diarrhoea, and is therefore protected by the mucus layer. As part of it, trefoil peptides (TFFs) are able to modify the visco-elastic properties of the mucus, protect against experimental ulceration, and promote repair of the epithelia. We investigated, using transient reporter gene assays and RT-PCR in the gastric carcinoma cell line MKN45 and colon carcinoma cell lines LS174T and HT29, whether ethanol and osmotic changes can modify transcriptional activity of TFFs. In a mild hypotonic environment (200 mosmol/l) all three TFF genes were up-regulated by at least a factor of 2. In hypertonic medium (400 mosmol/ll), TFF1 and TFF3 were down-regulated, whereas TFF2 was up-regulated by elevated concentrations of sodium or chloride in MKN45. Raising the osmolality by ethanol resulted in an up-regulation of TFF3 in both colon cell lines but not in the gastric cell line. We conclude that alteration in TFF gene expression is a response of gut epithelia to deal with osmotic forces and ethanol.


Subject(s)
Digestive System/metabolism , Ethanol/pharmacology , Gene Expression Regulation/drug effects , Growth Substances/genetics , Mucins , Muscle Proteins , Neuropeptides , Peptides/genetics , Digestive System/drug effects , Digestive System Physiological Phenomena , Down-Regulation , Humans , Proteins/genetics , Transcription, Genetic/drug effects , Trefoil Factor-1 , Trefoil Factor-2 , Trefoil Factor-3 , Tumor Cells, Cultured , Tumor Suppressor Proteins , Water-Electrolyte Balance
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