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PubMed2000

Effects of dietary calcium:phosphorus ratios on apparent absorption of calcium and phosphorus in the small intestine, cecum, and colon of pigs.

Thirty-two crossbred barrows were used to investigate the effects of dietary Ca:total P (tP) ratios in phytase-supplemented diets on the apparent absorption of P and Ca in the small intestine, cecum, and colon. Three Ca:tP ratio treatments (1.5:1, 1.3:1, or 1.0:1) were created by adjusting the amount of ground limestone added to the basal low-P grower (.39% tP including .07% added inorganic P) and finisher (.32% tP without added inorganic P) diets. All low-P ratio diets were supplemented with Natuphos phytase at 500 units/kg. A positive control diet without phytase supplementation contained adequate P and Ca to meet dietary requirements. At 123 kg, the pigs were slaughtered and the contents of ileum, cecum, and colon were collected. Lowering the dietary Ca:tP ratio in the diets containing phytase linearly increased (P < .01) the apparent absorption (% and g/d) of P in the small intestine, but Ca absorption was not affected. Pigs fed the low-P diet with a Ca:tP ratio of 1.0:1 had an apparent absorption (g/d) of P or Ca similar to that of pigs fed the control diet, which was adequate in Ca and P. Averaged across all diets, the apparent absorption of P was highest when measured at the cecum, and the apparent absorption of Ca was highest when measured at the colon. In conclusion, lowering the dietary Ca:tP ratio to 1.0:1 in a low-P diet containing phytase increased the apparent absorption of P in the small intestine. Furthermore, a significant amount of P was absorbed in the cecum.

6-Phytase↗
PubMed2003

New flow cytometric method to quantify the inhibition of enteropathogenic Escherichia coli adhesion by anti-adhesin antibodies.

BACKGROUND: Pathogenesis of enteropathogenic Escherichia coli (EPEC) infections can be divided in three stages. The first one is the intestinal colonization mediated by bacterial adhesins. The second and third stages are characterized by an intimate attachment of bacteria to the enterocytes. Little information is available on the specific immune response against EPEC. Here, we describe and validate a new approach to quantify the function of anti-EPEC adhesin antibodies (Abs). METHODS: We developed a new method to quantify the function of anti-adhesin Abs by flow cytometry. We used pEGFP-E22 (a rabbit EPEC E22 strain expressing the GFP protein) and HeLa cells. The adhesion of E22 bacteria to HeLa cells is mediated by AF/R2, the specific E22 adhesin. We performed short-time interaction (30 min) between pEGFP-E22 and HeLa cells. After extensive washes, 10,000 HeLa cells were acquired by flow cytometry and bacterial adhesion was quantified. Different sera were used to inhibit bacterial adhesion and recombinant MPB-Afr2G (Afr2G is the main AF/R2 subunit) was also tested in this system. RESULTS: We first verified that GFP expression by E22 did not modify bacterial adhesion. We then showed that this flow cytometry approach allowed easy quantification of bacterial adhesion and inhibition mediated by a specific anti-AF/R2 serum. Moreover, recombinant AF/R2 protein reversed the effect of the anti-AF/R2 serum. Finally, we validated our method using sera from E22 orally infected rabbits. We detected and quantified with this method functional specific anti-AF/R2 Abs in their sera. In addition, we correlated our results with an anti-AF/R2 enzyme-linked immunosorbent assay. CONCLUSIONS: We have developed a new method to detect and quantify specific anti-EPEC adhesin Abs by flow cytometry. This method is easy to use and highly reproducible. Its development could be extended to the search of specific anti-adhesin Abs in human EPEC infections.

Adhesins, Escherichia coli↗
PubMed1983

A case of ovarian dermoid cyst with malignant transformation perforated into the rectosigmoid colon and small intestine.

A 72-year-old female was admitted to hospital with diarrhea containing hair. The abdominal plain film showed teeth-like structures in the right lower quadrant. Barium enema and endoscopic examination were performed, and the results indicated a possible dermoid cyst perforated into the rectosigmoid colon and small intestine. The operation revealed a dermoid cyst of the right ovary perforated into the rectosigmoid colon and small intestine. Histopathologically, a dermoid cyst and in part well-differentiated squamous cell carcinoma invading the rectosigmoid and small intestine were demonstrated.

Aged↗
PubMed1996

Surgical resection for melanoma metastatic to the gastrointestinal tract.

OBJECTIVE: To evaluate the role of surgery in the survival of patients with melanoma metastatic to the gastrointestinal (GI) tract. DESIGN: Retrospective review. SETTING: Tertiary cancer center. PATIENTS: One hundred twenty-four potential surgical candidates with metastatic melanoma in the stomach, small intestine, colon, or rectum. MAIN OUTCOME MEASURES: Operative morbidity and mortality, relief of presenting symptoms, and median and 5-year survival. RESULTS: The median disease-free interval prior to diagnosis of GI tract metastasis was 23.2 months (range, 1-154 months). Patients typically presented with crampy abdominal pain, symptomatic mass, and/or occult GI tract blood loss. Of the 124 patients, 69(55%) underwent surgical exploration of the abdomen, 46 (66%) had curative resection, and 23 (34%) had a palliative procedure. There was only 1 operative death and 1 major operative complication; 67 (97%) of 69 surgical patients experienced postoperative relief of their presenting GI tract symptoms. The median survival in patients undergoing curative resection was 48.9 months, compared with only 5.4 months and 5.7 months in those undergoing palliative procedures and nonsurgical interventions, respectively. By multivariate analysis, the 2 most important prognostic factors for long-term survival were complete resection of GI tract metastases and the GI tract as the initial site of distant metastases. CONCLUSIONS: Almost all patients with melanoma and GI tract metastases can have palliation of symptoms by surgical intervention with minimal morbidity and mortality. The high 5-year survival rate associated with complete surgical resection of GI tract metastases indicates that surgery should be strongly considered for this subgroup of patients with melanoma and distant metastatic disease.

Female↗
PubMed1999

Characterization of the human 36-kDa carboxyl terminal LIM domain protein (hCLIM1).

We characterized a human cDNA clone encoding a 36-kDa carboxyl terminal LIM domain protein with a PDZ domain at the amino terminal. This full-length cDNA clone has a predicted open reading frame (ORF) of 329 amino-acid residues. The ORF of this cDNA encodes the human homolog of rat CLP36, and the putative protein is named human 36-kDa carboxyl terminal LIM domain protein (hCLIM1, nomenclature approved by the HUGO/GDB Nomenclature Committee). The hCLIM1 probe was used to hybridize with poly(A)+ RNA of various human tissues. Strong signals were detected in heart and skeletal muscle; moderate signals were detected in spleen, small intestine, colon, placenta, and lung; weaker levels were detected in liver, thymus, kidney, prostate, and pancreas; and no observable signals were detected in brain, testis, ovary, and peripheral blood leukocytes. The hCLIM1 gene was studied by fluorescence in situ hybridization (FISH), somatic cell hybrid analysis, and radiation hybrid mapping, and it is located at the human chromosome 10q26.

Amino Acid Sequence↗
PubMed1993

Tissue-specific expression of heat shock proteins of the mouse in the absence of stress.

The steady-state levels of four members of the heat shock proteins families (HSP84, HSC73, HSP71, and HSP25) were examined by immunoblot analysis of several different tissues of young and adult mice in the absence of stress. These hsps were detected in all tissues but their level was variable. The levels of HSC73 and HSP84 varied only slightly between different tissues in either young or adult mice, with the exception of skin where these hsps were found in reduced amounts. In contrasts, the stress-inducible member of the HSP70 family, HSP71, was found to be expressed in all tissues but in amounts which differed by as much as two orders of magnitude between tissues. In general, the levels of both HSP71 and HSP25 were found to be tissue dependent, with higher levels found in tissues such as stomach, intestine, colon and bladder, tissues which are exposed to toxic environmental or metabolic products, and which may concentrate these substances by water resorption and/or be exposed to them for longer periods. The levels of HSP71 and HSP25 were generally positively correlated both in young and adult mice although this correlation was not found in certain tissues such as kidney, testes, and bone. Tissues of young mice contained lower amounts of HSP25 and HSP71 than were found in the same tissues from adults. We conclude that hsps are expressed in all tissues of the mouse in the absence of stress and that some organs, particularly those exposed to potentially toxic metabolites, show a higher level of expression of HSP71 and HSP25.

Animals↗
PubMed2003

Familial and second primary pancreatic cancers: a nationwide epidemiologic study from Sweden.

Familial risk of pancreatic cancer has been mainly assessed through case-control studies based on reported but not medically verified cancers in family members. We used the nationwide Swedish Family-Cancer Database on 10.2 million individuals and 21,000 pancreatic cancers to calculate standardized incidence ratios (SIRs) and 95% confidence intervals (CIs) for pancreatic cancer in 0- to 66-year-old offspring of parents with pancreatic or other specified tumors. Additionally, SIRs for second primary pancreatic cancers were analyzed after any first neoplasm. SIRs for pancreatic cancer (1.68, 95% CI 1.16-2.35) and pancreatic adenocarcinoma (1.73, 95% CI 1.13-2.54) were increased when a parent presented with pancreatic cancer. The risk was not dependent on diagnostic age of offspring or parents. Pancreatic cancer was associated with parental lung, rectal or endometrial cancer and with melanoma. SIRs for pancreatic cancer were 10.01 and 7.96 among offspring who were diagnosed before age 50 years when parents were diagnosed with squamous cell and adenocarcinoma of the lung, respectively, before age 60 years. The population-attributable proportion of familial pancreatic cancer was 1.1%. Risks for second pancreatic cancers were increased in men and women after small intestinal, colon and bladder cancer. The degree of familial clustering for pancreatic cancer and its population-attributable proportion were lower than the data cited in the literature. Clustering of pancreatic cancer with sites presenting in hereditary nonpolyposis colorectal cancer was noted. The strong association of pancreatic and lung cancers is puzzling, and it remains unclear to what extent this represents familial sharing of smoking habits.

Adenocarcinoma↗
PubMed2004

Chemoprotective effect of caffeic acid phenethyl ester on promotion in a medium-term rat hepatocarcinogenesis assay.

Caffeic acid phenethyl ester (CAPE), a natural honeybee product exhibits a spectrum of biological activities including anti-microbial, anti-inflammatory, antioxidant and anti-tumoral actions. CAPE is also chemopreventive against intestinal, colon and skin cancer. Our aim was to extend the study of its chemoprotective features to the promotion of hepatocarcinogenesis. Male Wistar rats were subjected to a protocol under a modified promotion regimen of the resistant hepatocyte model. The altered hepatic foci (AHF) were quantitatively analyzed by histochemistry and image processing. When given during promotion, CAPE (20 mg/kg) decreased the expression of number and area gamma-glutamyl transpeptidase (GGT) positive AHF by 91% and 97%, respectively. When GGT expression was analyzed by RT-PCR, CAPE drastically decreased and prevented expression of almost all GGT transcripts at this stage of the carcinogenic process. Glutathione S-transferase placental form (GST-P), another protein marker for preneoplastic lesions was measured by Western blot and a decrease of 82% was observed. Additionally, we evaluated the effect of CAPE on the expression of nuclear factor NF-kappaB and found an 85% decrease in nuclear localization of the p65 subunit of NF-kappaB; however, their repressor, IkappaBalpha was not modified. Our results showed that CAPE given during promotion in hepatocarcinogenesis protects against induction of GGT-positive AHF, GST-P protein, GGT mRNA expression and translocation of p65. This phenomenon was independent of IkappaBalpha degradation.

Animals↗
PubMed1995

Oltipraz: clinical opportunities for cancer chemoprevention. p.

Oltipraz [4-methyl-5-(2-pyrazinyl)-1,2-dithiole-3-thione], originally developed as an antischistosomal agent, protects against chemical carcinogenesis in lung, trachea, forestomach, small intestine, colon, breast, skin, liver and urinary bladder in rodents. Oltipraz induces electrophile detoxication enzymes, resulting in diminished carcinogen-DNA adduct formation and reduced cytotoxicity, an important component of anticarcinogenic actions. Phase I trials of this drug have been recently conducted in the United States and indicate that the maximum tolerated dose is about 125 mg/day over a six-month period. Grade I/II dose-limiting toxicities included photosensitivity/heat intolerance, gastrointestinal, and neurologic toxicities. Ongoing studies are monitoring relationships between dose scheduling, drug plasma concentrations and pharmacodynamic action. Subsequent trials with this agent might most appropriately target individuals at high risk for occupational or environmental exposures to genotoxic carcinogens. Towards this end, a randomized, placebo-controlled Phase II study is planned for people at high risk for exposure to aflatoxins and development of hepatocellular carcinoma. Modulation of biomarkers reflecting the biologically effective dose of aflatoxin will serve as study endpoints.

Animals↗
PubMed1995

Chemoprevention by isothiocyanates.

Naturally occurring and synthetic isothiocyanates are among the most effective chemopreventive agents known. A wide variety of isothiocyanates prevents cancer in the rat lung, mammary gland, esophagus, liver, small intestine, colon, and bladder. Mechanistic studies have shown that this chemopreventive activity is due to favorable modification of phase I and phase II carcinogen metabolism, resulting in increased carcinogen excretion or detoxification and decreased carcinogen DNA interactions. Most studies reported that the isothiocyanate must be present at carcinogen exposure in order to effect tumorigenesis inhibition. Our studies focus on naturally occurring isothiocyanates phenethyl isothiocyanate (PEITC) and benzyl isothiocyanate (BITC) as lung cancer inhibitors. These studies employed the major lung carcinogens in tobacco smoke, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and benzo(a)pyrene (BaP). Combining chemopreventive agents that inhibit tumorigenesis by NNK and BaP in rodents may be effective in addicted smokers. PEITC inhibits lung tumor induction by NNK in F-344 rats and A/J mice, while BITC inhibits BaP-induced lung tumorigenesis in A/J mice; combining the two inhibits lung tumorigenesis by combined NNK and BaP in A/J mice. PEITC selectively inhibits metabolic activation of NNK in the rodent lung, while inducing glucuronidation of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), one of the major NNK metabolites. Thus, PEITC decreases DNA and hemoglobin adduct formation by NNK while increasing the amounts of NNAL and its glucuronide excreted in the urine. Presently available data indicate that non-toxic doses of PEITC can inhibit the metabolic activation and carcinogenicity of NNK in rat and mouse lung; BITC has similar effects on BaP activation and tumorigenicity in mouse lung. Thus, combinations of chemopreventive agents active against different carcinogens in tobacco smoke may be useful in the chemoprevention of lung cancer.

Animals↗
PubMed1986

Effect of liquid diet on nude mouse gastrointestinal tract.

The nude (athymic) mouse is currently used to study the effect of gastrin on cancer xenografts. We sought to develop a hypogastrinemia nude mouse model for use in evaluating the effect of hypogastrinemia on such xenografts. Thirty-six non-tumor-bearing nude mice were studied. Eighteen received a nutritionally complete liquid diet; eighteen received standard chow. Six mice from each group were weighed and killed (nonfasting) on days 2, 8, and 15. Mean serum gastrin levels (+/- SEM) for the control group were 118.7 +/- 7.5, 118.7 +/- 8.7, and 118.0 +/- 7.5 pg/ml on days 2, 8, and 15, respectively. Serum gastrin levels for the liquid diet group significantly decreased to 87.0 +/- 7.6, 88.0 +/- 9.7, and 66.7 +/- 9.6 pg/ml on the same days. Animals in both groups gained weight normally; there were no significant weight differences between the two groups at any point. No histological abnormalities were seen in stomach, small intestine, colon, cecum, liver, pancreas, spleen or kidney. However, the liquid diet group showed atrophic changes in colon: significant reductions in colon weight and RNA content on days 8 and 15, and significant reduction in colon protein content on day 8. This model of hypogastrinemia is reliable and inexpensive. The nonsurgical nature of the preparation allows excellent survival in this immunodeficient animal.

Animals↗
PubMed1995

Molecular cloning, sequence, and expression patterns of the human gene encoding CCAAT/enhancer binding protein alpha (C/EBP alpha).

The human gene encoding the transcription factor C/EBP alpha was isolated from an umbilical cord genomic library screened by low stringency hybridization. Two overlapping clones were characterized by restriction enzyme analysis and included 13.2 kb of the C/EBP alpha locus. The entire gene and 471 bp of the promoter were sequenced. The human C/EBP alpha gene is 2783 bp long and encodes a 356 amino acid long protein, which is the same in length as for rat C/EBP alpha. Compared to rat C/EBP alpha, there are two insertions of two amino acids and one deletion of four. The amino acid similarity between the two proteins is over 92%. The human C/EBP alpha gene was found to be expressed at the highest levels in placenta. High expression was also found in liver, lung, skeletal muscle, pancreas, small intestine, colon and in peripheral blood leukocytes. However, the expression was undetectable or very low in brain, kidney, thymus, testis and ovary. These results show that the human C/EBP alpha gene is expressed in a tissue restricted manner.

Amino Acid Sequence↗
PubMed1997

Molecular structure of the mouse CCK-A receptor gene.

We have cloned the mouse CCK-A receptor gene (Cckar), determined its nucleotide sequence, and analyzed its expression. The receptor protein is encoded in five exons distributed over 9 kb of genomic DNA. Intron/exon borders were determined by comparing the genomic nucleotide sequence with the mouse cDNA sequence obtained by reverse transcriptase polymerase chain reaction. RNase protection analysis of Cckar transcripts revealed the presence of a splice acceptor site 200 bp upstream of the translational start codon, indicating that the promoter is associated with a non-translated exon at an upstream site. The second coding exon contains a rarely used alternative splice site that would result in the production of a truncated, 48 amino acid protein. Cckar is widely expressed in the gastrointestinal system (pancreas, gallbladder, intestine, colon and stomach), as well as in brain and kidney.

Alternative Splicing↗
PubMed1997

Molecular cloning and tissue distribution of cDNA encoding a novel chemoattractant-like receptor.

With the application of a homology screening strategy, including PCR amplification and southern blot hybridization, a novel cDNA was cloned from rat liver and anterior pituitary libraries. It was found to encode a 371-amino acid protein which has the characteristics of a heptahelix receptor and shows structural identity to members of the chemoattractant receptor family. A primary receptor message of 3.5 kb size was identified by northern blot hybridization. This RNA species showed high expression in heart and lung, while expression was lower in small intestines, colon, kidney, liver, uterus, and in brain. Another larger RNA species of 6.3 kb appeared in heart and lung. In situ hybridization histochemistry performed on tissue from liver and kidney revealed a mainly vascular distribution of the receptor message.

Amino Acid Sequence↗
PubMed1998

Molecular cloning of a new sodium bicarbonate cotransporter cDNA from human retina.

Sodium bicarbonate cotransport is an electrogenic process and a principal regulator of pH. A recently cloned sodium bicarbonate cotransporter (NBC1) is predominantly expressed in kidney. The presence of several isoforms is suggested from functional studies. We have cloned a new member of this family from human retina, which was named hNBC2 (for human sodium bicarbonate cotransporter 2). The hNBC2 has 1018 amino acid residues with 10 hydrophobic transmembrane domains. A cluster of five N-glycosylation sites is present at the second extracellular loop. It has 53% amino acids identity with hNBC1 and 38% identity with anion exchanger 1. Thus, NBC2 is a new member of the anion exchanger superfamily. Northern blot of human tissues revealed its expression in many tissues such as testis, spleen, ovary, small intestine, colon, thymus, heart, and muscle (8.5 kb). The predominant expression at retina was documented with Northern blot of rat tissues. NBC2 may be a major regulator of extracellular pH of retina where light stimulation produces an extracellular alkalization and may contribute as a solute transporter to the prevention of retinal detachment.

Amino Acid Sequence↗
PubMed2001

Molecular characterization, tissue expression, and mapping of a novel Siglec-like gene (SLG2) with three splice variants.

The sialic acid binding immunglobulin-like lectin (Siglec) family is a recently described member of the immunoglobulin superfamily. Within the Siglec family, there exists a subgroup, which bears a high degree of homology with the molecule CD33 (Siglec-3), and has thus been designated the CD33-like subgroup of Siglecs. Members of this subgroup have been localized to chromosome 19q13.4. Through the positional candidate approach, we identified a novel potential member of this subgroup of Siglecs. We have characterized the complete genomic structure of this gene, determined its chromosomal localization, its homology to other members of the Siglec family, and its tissue expression profile. This new Siglec-like gene is comprised of 11 exons, with 10 intervening introns, and is localized 278 kb telomeric to Siglec-9 and 35 kb centromeric to Siglec-8 and on chromosome 19q13.4. The coding region consists of 2094 base pairs, and encodes for a putative 76.6 kDa protein. All Siglec-conserved structural features, including V-set domain, three C-set domains, transmembrane domain, ITIM and SLAM motifs, were found in this Siglec-like gene. Also, it has the conserved amino acids essential for sialic acid binding. The Siglec-like gene has 40-66% homology with members of the CD33-like subgroup, including Siglecs 5-9. Through RT-PCR we have examined the expression profile of this new gene in a panel of human tissues and found it to be primarily expressed in the bone marrow, spleen, brain, small intestine, colon, and spinal cord. We were also able to identify three different splice variants of the new gene. This gene may represent the latest novel member of the CD33-like subgroup of Siglecs, and, given its high degree of homology, it may also serve a regulatory role in the proliferation and survival of a particular hematopoietic stem cell lineage, as has been found for CD33 and Siglec-7.

Alternative Splicing↗
PubMed1994

Cloning, sequencing, and mapping of the human chromosome 14 heat shock protein gene (HSPA2).

A genomic clone for the human heat shock protein (HSP) 70 gene located on chromosome 14 was isolated and sequenced. The gene, designated HSPA2, has a single open reading frame of 1917 bp that encodes a 639-amino acid protein with a predicted molecular weight of 70,030 Da. Analysis of the sequence indicates that HSPA2 is the human homologue of the murine Hsp70-2 gene with 91.7% identity in the nucleotide coding sequence and 98.2% in the corresponding amino acid sequence. HSPA2 has less amino acid homology to other members of the human HSP70 gene family, 83.3% to the heat-inducible HSP70-1 gene and 86.1% with the human heat shock cognate gene HSC70. HSPA2 is constitutively expressed in most tissues, with very high levels in testis and skeletal muscle. Significant but lower levels are also expressed in ovary, small intestine, colon, brain, placenta, and kidney. A yeast artificial chromosome (YAC) clone containing HSPA2 (YAC741H4) that also contained the polymorphic marker D14S63 was identified. This 670-kb YAC was mapped to 14q24.1 by fluorescence in situ hybridization (FISH). Subsequent two-color FISH and genetic mapping placed HSPA2/D14S63 proximal to the markers D14S57 and D14S77.

Amino Acid Sequence↗
PubMed1998

Identification and molecular characterization of TM7SF2 in the FAUNA gene cluster on human chromosome 11q13.

In this report, the identification and molecular characterization of a novel gene, designated TM7SF2, is reported. This gene was found in the FAU neighboring area (FAUNA) to which other genes have been mapped previously. The FAUNA gene cluster is located at chromosome 11q13 between landmarks H4B and D11S2196E. The TM7SF2 gene contains eight coding exons, and their splice site consensus sequences are consistent with AG/GT rule. Northern blot analysis with a cDNA probe corresponding to TM7SF2 revealed varying expression levels of a 1.7-kb transcript in adult human heart, brain, pancreas, lung, liver, skeletal muscle, kidney, ovary, prostate, and testis, but no detectable expression in placenta, spleen, thymus, small intestine, colon (mucosal lining), or peripheral blood leukocytes. The open reading frame in the cDNA sequence codes for a protein of 590 amino acids that is rich in glycine (23%) and arginine (17%) residues in its amino-terminal half and contains seven transmembrane domains in its carboxy-terminal half. The transmembrane region of the putative TM7SF2 protein shows amino acid sequence similarity to those of the lamin B receptor and the C14/C24 sterol reductase.

Adult↗