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Parenteral Nutrition and Fasting Reduces Mucosal Addressin Cellular Adhesion Molecule-1 (MAdCAM-1) mRNA in Peyer's Patches of Mice

F. Enrique Gomez, PhD, Jinggang Lan, PhD, Woodae Kang, MD, Chikara Ueno, MD and Kenneth A. Kudsk, MD^*,

From the ^* Veterans Administration Surgical Services, William S. Middleton Memorial Veterans Hospital, and the Department of Surgery, University of Wisconsin–Madison College of Medicine and Public Health, Madison, Wisconsin

Correspondence: Kenneth A. Kudsk, MD, 600 Highland Avenue, H4/736 CSC, Madison, WI 53792-7375. Electronic mail may be sent to kudsk{at}surgery.wisc.edu.

Background: Mucosal addressin cellular adhesion molecule-1 (MAdCAM-1) in Peyer's patches (PP) is the gateway molecule for cellular migration into the mucosal immune system. Lack of enteral feeding during parenteral nutrition (PN) rapidly decreases PP MAdCAM-1, leading to drops in mucosal T and B cells and intestinal and respiratory IgA. We determined the molecular events associated with MAdCAM-1 mRNA and protein during PN (short and long term) and fasting (1 and 2 days). Methods: Experiment 1: Cannulated mice received PN for 8 hours (short-term PN, n = 6) or chow + saline (chow, n = 6). Experiment 2: Cannulated mice received PN (long-term PN, n = 4) or chow (n = 3) for 5 days. Experiment 3: Noncannulated chow mice were fasted for 1 and 2 days (n = 2/time). Total cellular RNA from the PP was quantified for MAdCAM-1 mRNA by real-time polymerase chain reaction (PCR). MAdCAM-1 protein was measured by Western blot. Results: PN rapidly down-regulated MAdCAM-1 gene expression. After 8 hours of PN with lack of enteral feeding, MAdCAM-1 mRNA levels dropped 20% (0.8-fold vs chow, p > .05); 5 days of PN reduced MAd-CAM-1 levels 64% (0.34-fold vs chow, p < .05). PN reduced MAdCAM-1 protein levels by 30% (chow: 329 ± 14 vs PN: 230 ± 35, p < .05) after 5 days. Fasting of uncannulated mice decreased MAdCAM-1 mRNA levels by 16% (0.84-fold, p < .05) at day 1 and 30% (0.7-fold, p < .05) by day 2 compared with chow. Conclusions: Both PN with lack of enteral feeding and fasting down-regulate MAdCAM-1 mRNA and protein levels in PP. The MAdCAM-1 changes are due to lack of enteral stimulation rather than toxic effects of PN.

Parenteral nutrition (PN) provides an important option for feeding when the oral or gastrointestinal routes are compromised, but clinical and experimental data demonstrate that PN is associated with more nosocomial infections, particularly pneumonia, compared with enteral feeding.^1–3 Our laboratory showed that PN alters mucosal immunity by decreasing lymphocytes in the gut- and mucosal-associated lymphoid tissue (GALT and MALT, respectively), lymphocyte CD4/CD8 ratio in the lamina propria (LP),⁴ gut Th-2 IgA-stimulating cytokines (IL-4 and IL-10),^5–7 and intestinal and respiratory IgA levels.^8–10 These changes create a defect in established respiratory defenses, which is reversible with chow feeding.^11–13

Naïve lymphocytes destined for GALT or MALT tissues enter the high endothelial venules (HEV) of the Peyer's patches (PP) via interaction with mucosal addressin cellular adhesion molecule-1 (MAdCAM-1). The naïve B and T lymphocytes migrate into the PP through interactions between MAdCAM-1 on the HEV and ₄β₇ and L-selectin expressed on the lymphocyte surface. The cells are subsequently distributed to the LP of intestinal and extraintestinal mucosal sites. PN significantly decreases PP MAdCAM-1 expression within 48 hours, with measurable changes within 8 hours of instituting PN.¹⁴ The reduction in MAdCAM-1 occurs simultaneously with reduction in the number of lymphocytes in the PP and LP. Selective antibody blockade of MAdCAM-1 recreates the PN-induced changes in LP lymphocytes.¹⁴ We hypothesized that the rapid changes observed in MAdCAM-1 protein were due to rapid molecular events occurring in the PP due to lack of enteral stimulation.

But there is a confounding variable in this experiment. We hypothesize that all changes are induced by the lack of enteral stimulation, but all experiments require that PN be administered because 3 days of fasting is lethal in these animals. It becomes unclear whether it is the lack of enteral nutrition or a toxic effect of the PN itself that causes alterations in MAd-CAM-1 expression. This confounding issue can be clarified by studying otherwise normal animals given a short-term fast. Therefore, the main objectives of this study were to (1) analyze the temporal effects of lack of enteral stimulation (with PN) on MAdCAM-1 gene expression in the PP by determining the MAdCAM-1 mRNA levels using a specific and sensitive real-time polymerase chain reaction (PCR) technique, (2) confirm that these changes have effects on MAdCAM-1 protein levels using Western blot techniques, and (3) study the effect of 1 and 2 days of lack of enteral stimulation by fasting (with no PN) on the levels of MAdCAM-1 mRNA in the PP in normal uncannulated mice.

	MATERIALS AND METHODS

Top MATERIALS AND METHODS RESULTS DISCUSSION

 
The Animal Care and Use Committee of the University of Wisconsin–Madison and the research committee of the Middleton Veteran's Administration Hospital approved all experimental protocols. Animals were housed in an American Association for Accreditation Laboratory Animal Care–accredited conventional facility. The environment was temperature and humidity controlled, with a daily 12-hour light-dark cycle.

Animals, Cannulation, and PN Feeding
Outbred male ICR mice 6 weeks old were purchased from Harlan (Madison, WI). Animals were fed ad libitum chow (Lab Diet 5001: PMI International, Brentwood, MO) and water for 2 weeks before protocol entry. Mice received catheters for IV infusion after the intraperitoneal injection of a ketamine (100 mg/kg body weight) and acepromazine maleate (5 mg/kg body weight) mixture. A silicone rubber catheter (0.012-inch inner diameter by 0.025-inch outer diameter; Baxter, Chicago, IL) was inserted into the vena cava through the right jugular vein. The distal end of the catheter was tunneled subcutaneously and exited the midpoint of the tail. The mice were partially immobilized by tail restraint to protect the catheter during infusion. This technique of infusion in the mouse has proved to be an acceptable method of nutrition support and does not produce physical or biochemical evidence of stress.¹⁵ Catheterized mice were immediately connected to an infusion apparatus, and 0.9% saline solution was infused at an initial rate of 4 mL/d, with ad libitum access to chow and water. After 2 days, mice were randomly assigned to continue on chow (control group) or to receive PN. The PN solution contained 4.1% amino acids, 34.3% glucose (4878 kJ/L), electrolytes, and multivitamins with a nonprotein calorie-to-nitrogen ratio of 743 kJ/g nitrogen. The PN group initially received 4 mL/d of PN solution and was advanced to a goal rate of 10 mL/d by the third day of feeding. The PN feedings met the calculated nutrition requirement of mice used in the present study, and their composition has been described in detail previously.⁴

Experimental Design
Experiment 1: Short-term PN. Twelve mice were cannulated, fed chow for 2 days to allow recovery from the stress of surgery, and then received PN (n = 6) or chow control (n = 6) for 8 hours before determining PP MAdCAM-1 expression by real-time PCR (mRNA levels). This timeframe was selected because we previously reported that MAdCAM-1 protein expression in PP declines after 4 hours of starting PN, reaching statistical significance after 48 hours.¹⁴

Experiment 2: Long-term PN. Seven mice were cannulated, fed chow for 2 days, and randomized to chow control (n = 3) or PN (n = 4) for 5 days to determine PP MAdCAM-1 expression by real-time PCR (mRNA levels) and by Western blot (protein levels).

Experiment 3: Fasting. To demonstrate that lack of enteral stimulation rather than the PN itself causes the observed reduction in MAdCAM-1 expression, mice were fasted for 1 (n = 2) or 2 (n = 2) days to determine MAdCAM-1 gene expression. Fasting was limited to 2 days only because a 3-day fast is lethal to most mice. Free access to drinking water was provided. For this experiment, only noncannulated mice were studied to avoid any alterations due to previous stress or weight loss from previous surgery. Uncannulated chow-fed mice (n = 2) served as controls.

Biochemical and Molecular Biological Methods
RNA isolation from the PP. The PP were excised at the times indicated, immediately frozen in liquid N₂ and stored at –80°C until analyzed. The PP were homogenized in 1 mL of Trizol reagent (Invitrogen, Life Technologies, Carlsbad, CA) with an Omni GLH General Laboratory Homogenizer (OMNI International, Marietta, GA) and the total cellular RNA was obtained by isopropanol precipitation. The integrity of the RNA was checked with 1% agarose gels stained with ethidium bromide (Promega, Madison, WI) by visualization of the 28S and 18S rRNA bands; its concentration was determined by UV at 260 nm and finally stored at –80°C until required.

cDNA synthesis. Total RNA was used as template to generate cDNA by reverse transcription under the following conditions: 2 µg of total RNA were incubated with 1 µg of oligo(dT)₁₅ (Promega) for 5 minutes at 70°C and kept on ice for 5 minutes. After the addition of 5x AMV-RT buffer (5 µL), ribonuclease inhibitor (40 U; RNasin, Madison, WI), dNTPs (5 µL of a 10 mmol/L stock solution), and 30 U of avian myeloblastoma virus reverse transcriptase (AMV-RT, Madison, WI) in a total volume of 25 µL, the mixture was incubated for 1 hour at 42°C. The enzyme was inactivated by incubating the samples for 10 minutes at 70°C and the cDNA was stored at –20°C until analysis.

Analysis of MAdCAM-1 gene expression by real-time PCR. Real-time PCR was performed using a Rotor-Gene 3000 thermal cycling system (Corbett Research, Australia) with SYBR-Green I (Molecular Probes, Eugene, OR) as fluorescent probe. After 45 cycles (20 seconds at 58°C, 15 seconds at 72°C, and 15 seconds at 95°C) a melting curve analysis of the products was done to assess for specific amplification.

The MAdCAM-1 forward (5'-AGT TAC TGT GCG CTG GAC CTT GGC TCC TGG CGA CCT GG-3') and reverse (5'-TCC TGG CGG CAC TGG AAC CAG CC-3') primers produced a single amplicon of 132 bp. The ratio of primers was optimized to achieve the maximum fluorescent signal with the lowest cycle threshold (Ct) value, and consisted of a combination of 300 nmol/L of each primer with 2 µL of a 1:10 dilution of the cDNA, in a cocktail mixture containing Platinum Quantitative PCR Super-Mix-UDG (Invitrogen, Life Technologies). The levels of MAdCAM-1 mRNA were normalized against β-actin mRNA levels as the housekeeping gene¹⁶; the β-actin primers (forward: 5'-CTA AGG CCA ACC GTG AAA AG-3'; reverse: 5'-ACC AGA GGC ATA CAG GGA CA-3') were used each at 300 nmol/L and produced a single amplicon of 104 bp.

Each sample was analyzed in triplicate to obtain the mean value of the Ct for MAdCAM-1 (Ct_MAdCAM-1) and for β-actin (Ct_β-actin). With these values, a difference in Ct (C_T) is obtained with the expression: C_T = Ct_MAdCAM-1 – Ct_β-actin.

The amount of MAdCAM-1 mRNA is normalized to β-actin by the comparative "change in cycle threshold" (C_T) method¹⁷ using the expression 2^–Ct, where Ct = [Ct_{(IV-parenteral nutrition)} – Ct_(chow)].

Western blot analysis. The PP were homogenized in 250 µL of 1x RIPA lysis buffer (Upstate, Lake Placid, NY) that contained proteases inhibitor cocktail (Sigma-Aldrich, St. Louis, MO). The crude homogenate was kept on ice for 1 hour and spun at 16,100 x g for 10 minutes at 4°C in a bench top Eppendorf 5415 refrigerated centrifuge. The supernatant was transferred to a clean tube, and the protein content was quantified with the dye-binding Bradford method.¹⁸ Twenty micrograms of protein were separated on a 10% PAGE under denaturing conditions (Tris 25 mmol/L, glycine 192 mmol/L, SDS 0.1%) at 150 V for 1 hour. After electrophoresis, the proteins were transferred to a 0.45 µm PVDF membrane (Immobilon-P, Millipore Corporation, Bedford, MA) at 4°C, 80 V for 45 minutes in transfer buffer (Tris 48 mmol/L, glycine 39 mmol/L, methanol 20%). The membrane was washed 3 times for 5 minutes each wash with TBS-Tween (Tris-buffered saline plus 0.05% Tween-20) and was blocked for 3 hours with blocking solution (5% nonfat dry milk in TBS-Tween) with constant agitation at room temperature. The membrane was incubated overnight at 4°C with constant agitation with the primary antibody, a rat monoclonal anti-mouse MAdCAM-1 (clone MECA-367, Pharmigen BD-Biosciences) diluted 1:500 in blocking solution. The membrane was washed as before and incubated 1 hour at room temperature with the secondary antibody: goat anti-rat IgG (H&L)–HRP conjugate (Serotec, Cambridge, MA) diluted 1:20,000 in blocking solution.

View larger version (9K): [in this window] [in a new window]   FIGURE 1. MAdCAM-1 mRNA measured by real-time polymerase chain reaction (PCR) decreased from chow control levels by 8 hours after starting PN and reached statistical significance (p < .05) at day 5.

Statistical Analysis
The amount of MAdCAM-1 mRNA is expressed relative to that of β-actin mRNA as a "n-fold" change using the expression 2^–Ct. By definition, the value of Ct is zero for the control group (Ct_chow –Ct_chow), and 2⁰ = 1.0, that is, the normalized value of the control group. Differences were considered significant (p < .05) if the range in the PN group did not include this value of 1.0, because this value means "no change" in respect to the chow group.

The levels of MAdCAM-1 protein that were determined by Western blot were analyzed with the Student's t-test, with p .05 considered statistically significant.

	RESULTS

Top MATERIALS AND METHODS RESULTS DISCUSSION

 
Body Weight
The body weight of the animals used in experiments 1, 2, and 3 is noted in Table I. There were no significant differences in body weight between any of the groups.

Effects of PN on MAdCAM-1 mRNA Levels (Experiments 1 and 2)
Figure 1 shows the results of the qPCR analysis for the expression of MAdCAM-1 in the chow and PN groups.

View larger version (16K): [in this window] [in a new window]   FIGURE 2. Western blot of MAdCAM-1 in PP of mice fed chow and PN for 5 days. A, Coomassie-stained gel showing that equal amounts of protein (20 µg/lane) were loaded. Lanes 1 and 2, chow-fed mice; lanes 3 and 4, PN-fed mice. B, Western blot analysis showing that the antibody MECA-367 detects a single band of 52 kDa corresponding to MAdCAM-1, which is reduced in the PN group. C, Graphic representation of the levels of MAdCAM-1 protein in the chow (white bar, 230 ± 35; n = 3) and the PN (black bar, 329 ± 14; n = 4) groups.

Experiment 2: Long-term (5 days) PN resulted in a significant decrease in MAdCAM-1 mRNA levels of 64% (0.34-fold; range, 0.30–0.37; p < .05) compared with the chow group.

Effects of PN on MAdCAM-1 Protein Levels (Experiment 2)
Figure 2 shows the MAdCAM-1 protein levels determined by Western blot. Equal amounts of protein (20 µg) were separated by electrophoresis and transferred to a PVDF membrane. The Coomassie-stained gel analysis (Figure 2A) confirmed that equal amounts of protein were loaded. Western blot analysis showed a single band of 52 kDa for MAdCAM-1 (Figure 2B). Densitometric analysis of the resulting bands confirmed that PN feeding significantly reduced MAd-CAM-1 protein compared with the chow group (230 ± 35 vs 329 ± 14, p < .05; Figure 2C).

View larger version (10K): [in this window] [in a new window]   FIGURE 3. MAdCAM-1 mRNA measured by real-time polymerase chain reaction (PCR) decreased significantly from chow controls after 24 and 48 hours of fasting (p < .05).

	DISCUSSION

Top MATERIALS AND METHODS RESULTS DISCUSSION

 
The gastrointestinal tract performs the vital functions of digestion and absorption of nutrients while protecting the body from harmful bacterial products, biologically active dietary peptides produced during digestion,¹⁹ and environmental toxins and pollutants. When the gastrointestinal tract function is compromised for prolonged periods of time, PN may be a life-sustaining therapy. However, failure to deliver enteral nutrients produces immunologic gaps in mucosal defenses resulting in infectious complications.^20–22

Under normal feeding conditions, naïve T and B cells express surface ₄β₇ and L-selectin. These molecules interact with MAdCAM-1 and other integrins such as ICAM-1 on the HEV of the PP. After initial interaction and attachment, chemokines in the PP stimulate migration of cells into the PP, where they are sensitized and migrate to the mesenteric lymph node for continued stimulation, proliferation, and maturation before distribution via the vascular system to mucosal immune sites throughout the body.

Our group showed that MAdCAM-1 expression in PP significantly decreases within 48 hours of initiating PN using a dual-labeled monoclonal antibody technique, with measurable changes within 4–8 hours.¹⁴ Experimentally, monoclonal antibody blockade of endothelial MAdCAM-1, lymphocyte ₄β₇, or lymphocyte L-selectin reproduces these cellular changes throughout GALT. ICAM-1 blockade has no effect.²³ Consistent with the rapid MAdCAM-1 depression in PP, the current work demonstrates a rapid decrease in transcription of the MAdCAM-1 gene measurable within 8–24 hours of stopping enteral stimulation, with further depression at 5 days.

The full-length wild-type murine MAdCAM-1 protein consists of 384 amino acids and comprises 6 regions: 4 extracellular domains, 1 transmembrane region, and 1 short cytoplasmic tail.²⁴ The first 2 domains share homology with ICAM-1 and VCAM-1 (vascular cell adhesion molecule-1) respectively, whereas the third domain is mucin-rich and supports L-selectin-mediated lymphocyte rolling.²⁵ The fourth domain bears homology to the C2 domain of IgA1. The gene for MAdCAM-1 is located on chromosome 10 and consists of 5 exons: 1 of these, exon 4, encodes for both the mucin-rich region and the IgA-like domain.²⁶ Alternative splicing of the MAdCAM-1 gene, which normally occurs in the PP, produces 2 species of mRNA, one of about 1.6 kb and another of 0.8 kb.^24,26,27 The shorter transcript, which is the result of the deletion of exon 4, is predicted to produce a truncated form of MAdCAM-1 of only 240 amino acids long, lacking both the mucin-rich and the IgA-like domains.^26,27 This is important because the deletion of both domains affects the capacity of MAdCAM-1 to support primary lymphocyte adhesion through its interaction with L-selectin and with ₄β₇ ^25,26 integrin. As noted in our previous work, blockade of L-selectin significantly impairs T- and B-cell migration into PP.²³

In this study, the set of primers used in the real-time PCR technique were specifically designed to amplify a region spanning 132 bp that is found only in exon 4. In this way, we were able to determine the long form of the MAdCAM-1 mRNA and, by extension, of the full-length fully functional protein. Any other exon in the MAdCAM-1 gene targeted for its analysis using realtime PCR would not distinguish the long from the short MAdCAM-1 mRNAs, because all other exons are found in both transcripts. This would interfere with the interpretation of the results obtained by this technique. Our real-time PCR technique was sensitive enough to detect changes in the MAdCAM-1 mRNA levels as early as 8 hours (although not statistically significant) after starting parenteral feeding.

Because alterations in mRNA do not guarantee alterations in protein expression, the Western blot results confirm decreased expression of MAdCAM-1 protein in PP of PN-fed mice. This is consistent with and extends our previous work, which showed a significant decrease in PP's MAdCAM-1 using the dual-labeled monoclonal antibody technique.¹⁴ In addition, the 24- and 48-hour fasting experiments confirmed that the decrease in MAdCAM-1 mRNA levels can be attributed to the lack of enteral stimulation rather than an inhibitory effect of the PN solution.

Our results indicate that PN and fasting down-regulate MAdCAM-1 gene expression both at the mRNA and protein levels. It remains unclear how enteral stimulation signals active transcription of the intact MAdCAM-1 gene. It is not secondary to the enteric nervous system or the enterocyte amino acid fuel, glutamine, because our previous work showed that neither bombesin, a neuropeptide analogous to gastrin-releasing peptide in humans, nor glutamine exerts any effect on MAdCAM-1 expression during parenteral feeding.²⁸ These results support our hypothesis that changes in MAdCAM-1 are due to lack of enteral stimulation and are not due to toxic effects of PN.

The study was supported by National Institutes of Health grant R01 GM53439.

Received for publication December 21, 2005. Accepted for publication August 15, 2006.

1. Kudsk KA, Croce MA, Fabian TC, et al. Enteral versus parenteral feeding: effects on septic morbidity after blunt and penetrating abdominal trauma. Ann Surg. 1992;215:503 –513.[Web of Science][Medline] [Order article via Infotrieve]
2. Varga P, Griffiths R, Chiolero R, et al. Is parenteral nutrition guilty? Intensive Care Med.2003; 29:1861 –1864.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
3. Gramlich L, Kichian K, Pinilla J, Rodych NJ, Dhaliwal R, Heyland DK. Does enteral nutrition compared to parenteral nutrition result in better outcomes in critically ill adult patients? A systematic review of the literature. Nutrition.2004; 20:843 –848.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
4. Li J, Kudsk KA, Gocinski B, Dent D, Glezer J, Langkamp-Henken B. Effects of parenteral and enteral nutrition on gut-associated lymphoid tissue.J Trauma. 1995;39:44 –52.[Web of Science][Medline] [Order article via Infotrieve]
5. Kudsk KA, Wu Y, Fukatsu K, et al. Glutamine-enriched total parenteral nutrition maintains intestinal interleukin-4 and mucosal immunoglobulin A levels. JPEN J Parenter Enteral Nutr.2000; 24:270 –275.[Abstract/Free Full Text]
6. Zarzaur BL, Wu Y, Fukatsu K, Johnson CD, Kudsk KA. The neuropeptide bombesin improves IgA-mediated mucosal immunity with preservation of gut interleukin-4 in total parenteral nutrition-fed mice. Surgery.2002; 131:59 –65.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
7. Fukatsu K, Kudsk KA, Zarzaur BL, Wu Y, Hanna MK, DeWitt RC. TPN decreases IL-4 and IL-10 mRNA expression in lipopolysaccharide stimulated intestinal lamina propria cells but glutamine supplementation preserves the expression. Shock. 2001;15:318 –322.[Web of Science][Medline] [Order article via Infotrieve]
8. Hanna MK, Zarzaur BL, Fukatsu K, et al. Individual neuropeptides regulate gut-associated lymphoid tissue integrity, intestinal immunoglobulin A levels, and respiratory antibacterial immunity. JPEN J Parenter Enteral Nutr. 2000;24:261 –269.[Abstract/Free Full Text]
9. Janu P, Li J, Renegar KB, Kudsk KA. Recovery of gut-associated lymphoid tissue and upper respiratory tract immunity after parenteral nutrition. Ann Surg.1997; 225:707 –717.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
10. Johnson CD, Kudsk KA, Fukatsu K, Renegar KB, Zarzaur BL. Route of nutrition influences generation of antibody-forming cells and initial defense to an active viral infection in the upper respiratory tract. Ann Surg. 2003;237:565 –573.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
11. Kudsk KA, Li J, Renegar KB. Loss of upper respiratory tract immunity with parenteral feeding. Ann Surg.1996; 223:629 –638.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
12. Renegar KB, Kudsk KA, DeWitt RC, Wu Y, King BK. Impairment of mucosal immunity by parenteral nutrition: depressed nasotracheal influenza-specific secretory IgA levels and transport in parenterally fed mice. Ann Surg. 2001;233:134 –138.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
13. Renegar KB, Johnson CD, DeWitt RC, et al. Impairment of mucosal immunity by total parenteral nutrition: requirement for IgA in murine nasotracheal anti-influenza immunity. J Immunol.2001; 166:819 –825.[Abstract/Free Full Text]
14. Ikeda S, Kudsk KA, Fukatsu K, et al. Enteral feeding preserves mucosal immunity despite in vivo MAdCAM-1 blockade of lymphocyte homing. Ann Surg. 2003;237:677 –685.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
15. Sitren HS, Heller PA, Bailey LB, Cerda JJ. Total parenteral nutrition in the mouse: development of a technique. JPEN J Parenter Enteral Nutr. 1983;7:582 –586.[Abstract/Free Full Text]
16. Bas A, Forsberg G, Hammarstrom S, Hammarstrom ML. Utility of the housekeeping genes 18S rRNA, β-actin and glyceraldehyde-3-phosphate-dehydrogenase for normalization in real-time quantitative reverse transcriptase-polymerase chain reaction analysis of gene expression in human T lymphocytes. Scand J Immunol.2004; 59:566 –573.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
17. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2^-Ct method.Methods. 2001;25:402 –408.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
18. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem.1976; 72:248 –252.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
19. Zaloga GP, Siddiqui RA. Biologically active dietary peptides.Mini Rev Med Chem. 2004;4:815 –821.[Web of Science][Medline] [Order article via Infotrieve]
20. Kudsk KA, Minard G, Croce MA, et al. A randomized trial of isonitrogenous enteral diets following severe trauma: an immune-enhancing diet (IED) reduces septic complications. Ann Surg.1996; 224:531 –543.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
21. Moore FA, Moore EE, Jones TN, McCroskey BL, Peterson VM. TEN versus TPN following major abdominal trauma: reduced septic morbidity. J Trauma. 1989;29:916 –922.[Web of Science][Medline] [Order article via Infotrieve]
22. Moore FA, Moore EE, Jones TN. Benefits of immediate jejunostomy feeding after major abdominal trauma: a prospective randomized study. J Trauma. 1986;26:874 –881.[Web of Science][Medline] [Order article via Infotrieve]
23. Reese S, Kudsk KA, Genton L, Ikeda S. L-selectin and 4β7 integrin, but not ICAM-1, regulate lymphocyte distribution in gut-associated lymphoid tissue of mice. Surgery.2005; 137:209 –215.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
24. Briskin MJ, McEvoy LM, Butcher EC. MAdCAM-1 has homology to immunoglobulin and mucin-like adhesion receptors and to IgA1.Nature. 1993;363:461 –464.[CrossRef][Medline] [Order article via Infotrieve]
25. Berg EL, McEvoy LM, Berlin C, Bargatze RF, Butcher EC. L-selectin-mediated lymphocyte rolling on MAdCAM-1. Nature.1993; 366:695 –698.[CrossRef][Medline] [Order article via Infotrieve]
26. Sampaio SO, Li X, Takeuchi M, et al. Organization, regulatory sequences and alternatively spliced transcripts of the mucosal addressin cell adhesion molecule-1 (MAdCAM-1) gene. Immunology.1995; 155:2477 –2486.
27. Schiffer SG, Day E, Latanision SM, Tizard R, Osborn L. An alternately spliced mRNA encoding functional domains of murine MAdCAM-1.Biochem Biophys Res Commun.1995; 216:170 –176.[CrossRef][Web of Science][Medline] [Order article via Infotrieve]
28. Zarzaur BL, Ikeda S, Johnson CD, Le T, Sacks G, Kudsk KA. Mucosal immunity preservation with bombesin or glutamine is not dependent on mucosal addressin cell adhesion molecule-1 expression. JPEN J Parenter Enteral Nutr. 2002;26:265 –270.[Abstract/Free Full Text]

Journal of Parenteral and Enteral Nutrition, Vol. 31, No. 1, 47-52 (2007)
DOI: 10.1177/014860710703100147


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