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M E Zenilman

Publications and source records attributed to M E Zenilman.

At least 19 recordsLinked to original sources
PubMed2004

Targeted inhibition of gene expression of pancreatitis-associated proteins exacerbates the severity of acute pancreatitis in rats.

BACKGROUND: Pancreatitis-associated protein (PAP) is a secretory protein not normally expressed in healthy pancreas but highly induced during acute pancreatitis. While PAP has been shown to be anti-bacterial and anti-apoptotic in vitro, its definitive biological function in vivo is not clear. METHODS: To elucidate the function of PAP, antisense oligodeoxyribonucleotides (AS-PAP) targeting all three isoforms of PAP were administered via intrapancreatic injections (5 mg kg day, 2 days) to rats prior to induction of pancreatitis. RESULTS: Severity of pancreatitis and cytokine gene expression in peripheral blood mononuclear cells (PBMC) were evaluated. Administration of AS-PAP, but not the scrambled oligodeoxyribonucleotide (SC-PAP) control, reduced pancreatitis-induced PAP expression by 55.2 +/- 6.4%, 44.0 +/- 8.9%, and 38.9 +/- 10.7% for PAP isoforms I, II, and III, respectively, compared to saline-treated controls (P < 0.05 for all). Inhibition of PAP expression significantly worsened pancreatitis: serum amylase activity, pancreas wet weight (reflecting edema), and serum C-reactive protein levels all increased in AS-PAP-treated animals compared to SC-PAP-treated controls (by 3.5-, 1.7-, and 1.7-fold, respectively; P < 0.05 for all). Histopathologic evaluation of pancreas revealed worsened edema, elevated leukocyte infiltration, and fat necrosis after AS-PAP treatment. Gene expressions of IL-1 microm and IL-4 were significantly higher in PBMC isolated from AS-PAP-treated rats compared to SC-PAP controls. CONCLUSION: This is the first in vivo evidence indicating that PAP mediates significant protection against pancreatic injury. Our data suggest that PAP may exert its protective function by suppressing local pancreatic as well as systemic inflammation during acute pancreatitis.

Acute Disease↗
PubMed2000

A recombinant rat regenerating protein is mitogenic to pancreatic derived cells.

Pancreatic regenerating protein (reg I) is expressed in acinar cells and is mitogenic to beta- and ductal cells. Isolation of large amounts of endogenous reg I for in vivo and in vitro experiments has been difficult. The aim of this study was to develop a recombinant protein and determine its bioactivity on rat pancreatic derived cells. cDNA of the rat reg I coding region was created with unique BamHI flanking sequences using reverse transcriptase PCR. The coding region was then cloned into a bacterial expression vector in which expression is controlled by a T7 promoter. After transformation into the Escherichia coli strain B21(DE3) and induction by isopropyl-beta-d-thiogalactopyranoside, a fusion protein of 24 kDa in size, named reg-PET, was noted in the bacterial lysate. This protein contained a polyhistidine and S-peptide sequence to facilitate isolation and identification, respectively. This protein was purified using affinity chromatography, and identity was confirmed with gel electrophoresis and Western analysis. The reg-PET protein was mitogenic to both ARIP and RIN cells, rat pancreatic ductal and beta-cell lines, respectively. Antibodies raised to the protein reacted against rat reg I in pancreas. The purified recombinant reg I fusion protein, like endogenous reg I, is mitogenic to pancreatic derived cells. It is more potent than reg I isolated from pancreatic tissue. This protein can be isolated rapidly, easily, and with a high amount of purity.

Amino Acid Sequence↗
PubMed2000

Comparison of reg I and reg III levels during acute pancreatitis in the rat.

OBJECTIVE: To study alterations of serum levels of the pancreatic reg family of proteins in two models of acute pancreatitis. SUMMARY BACKGROUND DATA: The pancreatic reg family of proteins is expressed in the acinar pancreas. Reg I (pancreatic stone protein, PSP) and reg III (pancreatitis-associated protein, PAP) are induced after the onset of acute pancreatitis, and both have been proposed as potential markers of pancreatitis. METHODS: Pancreatitis was induced in rats by either retrograde infusion of sodium taurocholate or by direct trauma. Serum samples were obtained daily for 4 days after the procedure, and the animals were then killed. Twelve animals underwent sham procedure and six underwent daily analysis without surgery. Levels of reg I/PSP and reg III/PAP were estimated by enzyme-linked immunosorbent assay. RESULTS: Reg III/PAP levels increased significantly the first day after induction of both types of pancreatitis and rapidly returned to baseline in all survivors. Even animals who received retrograde infusion of saline showed a mild increase in reg III/PAP on the first day, whereas control animals that did not undergo surgery showed no variations. Reg I/PSP serum levels remained unchanged throughout all experimental periods. Postinjury reg III/PAP levels significantly correlated with severity of the pancreatic injury and animal survival; reg I/PSP levels did not. CONCLUSION: After induction of pancreatitis, serum levels of reg I and III protein differ significantly. Reg III/PAP levels are a sensitive marker of pancreatic injury and early in the disease may be a useful prognostic indicator for disease severity.

Acute Disease↗
PubMed2000

Use of pancreaticogastrostomy for pancreatic reconstruction after pancreaticoduodenectomy.

After resection of the head of the pancreas, the distal remnant is typically telescoped into the jejunum. Recently, pancreaticogastrostomy has re-emerged as a useful alternative, as the anastomosis is easy and without tension. The results of pancreaticogastrostomy in 10 consecutive patients is reviewed, as is the literature of both the technique and the physiology of the procedure. In the current series, mean +/- SEM age was 65 +/- 2.3 years. Extended pancreaticoduodenectomy was performed in two patients, pylorus-preserving in eight. Mean tumor size was 3.9 +/- 1.1 cm (range, 1.5-7.5 cm), mean operative time was 6.5 +/- 0.5 hours. Intraoperatively, 7.6 +/- 0.8 L of fluid was given, only two patients were transfused. The mean length of stay was 9.4 +/- 1.8 days. There were no anastomotic leaks, no deaths, and two patients developed temporary gastric ileus. There are now 841 pancreaticogastrostomies reported in the literature, with a leak rate of 3.1% and a death rate of 2.6%. Pancreaticogastrostomy is easy to perform, safe, and useful even after extended Whipple.

Anastomosis, Surgical↗
PubMed1998

Effect of reg protein on rat pancreatic ductal cells.

The pancreatic regenerating gene (reg I) is expressed in the exocrine pancreas and is involved in islet regeneration. Reg I protein has been shown to be mitogenic to beta- and ductal cell lines, but not mature islets. In this study, we tested the effect of two isolates of reg I on primary cultures of ductal cells. Rat pancreatic ductal cells were isolated by collagenase digestion and isolated colonies were maintained in culture. The cells were proven to be ductal in origin by their morphology and by immunofluorescent staining with epithelial markers. Reg I was isolated from human pancreatic extracts or from the rat acinar cell line AR42J by sequential ammonium sulfate precipitation and acid precipitation. Cells were cultured with doses of reg I for 72 h, pulsed with 10 microM bromodeoxyuridine (BrdU) for 2 h. After fixation, nuclei were double-stained with propidium iodide and BrdU monoclonal antibody. The percentages of nuclei positive for BrdU were calculated from at least five colonies per group. A 10-nM concentration of human reg I increased BrdU incorporation by 2.3-fold over controls, rat reg I increased it by 1.4-fold (p < 0.05). When compared to their effects on the ductal cell line ARIP, both human and rat reg I were 100 times more potent on the primary cultures of ductal cells. We conclude that human and rat reg I proteins are mitogenic to primary cultures of ductal cells. Although principally a product of the acinar cell, reg I appears to be a stimulus of ductal cell growth and, in this fashion, may modulate the expansion of the pancreatic ductal population during islet regeneration.

Animals↗
PubMed1998

Characteristics of rat pancreatic regenerating protein.

BACKGROUND: The pancreatic regenerating (reg) gene is an acinar cell product involved in islet formation and maintenance. Human reg protein is mitogenic to pancreatic beta and ductular cells, and its amino acid sequence predicts it to be a calcium-dependent lectin. METHODS: We studied the biologic activity and cellular localization of rat reg I isolated from the acinar cell line AR42J and the lectin properties of reg from AR42J and pancreatic juice. Bioactivity was assayed by mitogenesis on the ductular cell line ARIP. Cellular localization was determined by differential centrifugation. Lectin properties were assessed by affinity chromatography. RESULTS: Reg protein from crude AR42J cellular lysate and purified reg protein from AR42J induced thymidine incorporation to ARIP. Conditioned medium from AR42J and co-culture of AR42J with morphologically distinguishable ARIP, however, failed to induce mitogenesis. Reg protein was localized within the vesicle fraction of the cell and was not membrane bound. Affinity chromatography revealed that reg protein did not bind to mannose or galactose in the presence or absence of calcium. In pancreatic juice a previously undescribed mannose-binding protein was discovered at 25,000 to 30,000 daltons. CONCLUSIONS: We conclude that reg produced in the acinar cell line AR42J is biologically active but not efficiently secreted, even though it localized within the cellular vesicles. Despite predictions based on its amino acid sequence, it does not appear to be a calcium-dependent lectin.

Animals↗
PubMed1997

Pancreatic reg gene expression is inhibited during cellular differentiation.

BACKGROUND AND OBJECTIVE: Factors that control pancreatic regenerating (reg I) gene expression are unknown, but it is believed that its expression may correspond with cellular differentiation. The authors recently demonstrated that reg I is expressed in AR42J, a rat acinar cell line whose state of differentiation can be modulated by dexamethasone. They used this line to study reg I expression during cellular proliferation and differentiation. METHODS: After treatment of cells with 10 nmol/L dexamethasone, proliferation was assayed by thymidine incorporation; differentiation by expression of elastase I mRNA. Reg I mRNA levels were measured using a rat reg I cDNA probe, and reg I protein levels assayed by enzyme-linked immunosorbent assay of cellular lysates with a polyclonal antibody. The effect of gastrin, cholecystokinin and glucagon on reg I expression was also studied. RESULTS: When compared with controls, treatment with dexamethasone caused thymidine incorporation to decrease and elastase mRNA levels to increase. Reg I mRNA decreased from controls of 100 +/- 16% to 40 +/- 18% (p < 0.05), and reg I protein levels decreased as well. Gastrointestinal hormones had no significant effect on either elastase or reg I gene expression. CONCLUSIONS: Expression of reg I inversely correlates with the level of cellular differentiation, can be modulated via the glucocorticoid receptor, and is a potential marker of gastrointestinal epithelial differentiation. Despite its presence within a pancreatic acinar cell line, reg I gene expression is not modulated by gastrointestinal hormones.

Animals↗
PubMed1997

Insulin-like growth factor I receptor messenger RNA in the colon is unchanged during neoplasia.

The expression of growth factor receptor messenger RNA is difficult to quantitate due to low copy number. We describe a quantitative polymerase chain reaction that rapidly, reproducibly assays expression of human insulin-like growth factor I (IGF-I) receptor mRNA from small, biopsy-sized, specimens of tissue. This was then clinically applied to surgically resected specimens of colon. Total RNA was isolated from normal colonic mucosa and documented tumors from 4 patients undergoing resection. The mRNA was first reverse-transcribed with an oligomer bearing a complementary sequence specific for the mRNA at its 3' end, and a sequence complementary to an intervening intron of the IGF-I receptor gene at the 5' end. Competitive PCR was then performed in the presence of the cDNA product and exogenously added genomic DNA, with an upstream primer complementary to the exon sequence of the gene of interest and a downstream primer complementary to the intron sequence that was tagged to the cDNA. The genomic DNA was used as the internal standard. To calculate the number of copies of mRNA per microgram total RNA, a standard curve was used. No difference was noted in IGF-I receptor expression between neoplastic and normal colonic mucosa. This quantitative PCR is accurate, rapid, and requires very small amounts of tissue. Potential uses are in determining genetic expression of growth factor receptors or putative tumor markers preoperatively from small samples obtained during diagnostic colonoscopy or from samples obtained at surgery.

Colon↗
PubMed1997

Laparoscopic cholecystectomy: applicability in the geriatric population.

Elderly patients with cholelithiasis are more likely than nonelderly patients to present with an acute complication of gallstone disease such as acute cholecystitis (AC), gallstone pancreatitis (GSP), or common bile duct stones (CBDS). These acute complications may make laparoscopic cholecystectomy (LC) more hazardous, with a potential increase in perioperative morbidity or need for open conversion. The applicability of LC in the geriatric population is, therefore, unclear. We reviewed 283 consecutive patients undergoing attempted LC. Patients were classified as presenting with complicated (AC, GSP, or CBDS) or uncomplicated gallstone disease. Elderly patients were significantly more likely than younger patients to present with AC (40% versus 18%), GSP (19% versus 6%), and CBDS (21% versus 5%) (all P < 0.05). Elderly patients with chronic, uncomplicated gallstone disease (n = 20) and nonelderly patients with uncomplicated disease (n = 159) had similar open conversion rates (5% and 7%, respectively). In contrast, the open conversion rate in elderly patients with complicated gallstone disease (n = 42) was significantly higher (50%) compared to nonelderly patients with complicated disease (n = 62; rate, 16%; P < 0.05). Perioperative morbidity and length of stay were also significantly increased in the elderly group, primarily due to the high percentage of elderly patients with complicated disease. These results suggest that elderly patients with uncomplicated gallstone disease appear to be excellent candidates for LC, and this should be considered before complicated disease develops. Conversely, early conversion or planned open cholecystectomy may be warranted in the elderly presenting with acute complications of cholelithiasis.

Aged↗
PubMed1996

Pancreatic regeneration (reg) gene expression in a rat model of islet hyperplasia.

BACKGROUND: After pancreatectomy, regeneration of acinar and islet elements occurs. Recent data from a model of islet hyperplasia in the hamster suggested that induction of a local pancreatic factor stimulates islet formation. We postulate that the reg gene may be this factor. METHODS: We studied reg expression during induction of islet growth by using a similar model in the rat. Rats underwent surgical wrapping of the splenic lobe of the pancreas or sham operation. RESULTS: At 2 days ductular proliferation and immunohistochemical evidence of insulin within ductular epithelia were evident in the wrapped lobe. By 14 and 56 days islet number per square millimeter increased 63% and 43%, respectively. Reg mRNA levels, measured by Northern blot analysis with a rat reg cDNA probe (n = 5), increased 300% at 2 days in the wrapped lobe and decreased to that of unwrapped controls by 14 days. In situ hybridization showed localization of reg to the acinar cells. In unwrapped gastric lobes of animals who underwent surgical wrapping of the splenic lobe, no change in islet number per square millimeter or reg gene expression was noted. CONCLUSIONS: Surgical wrapping of the pancreatic splenic lobe induces local reg gene expression that is temporally associated with duct cell hyperplasia. This is followed by islet formation within the wrapped lobe. Reg may play a role in the induction of new islets from ductular precursors and in maintenance of normal islet function.

Amylases↗
PubMed1996

Pancreatic thread protein is mitogenic to pancreatic-derived cells in culture.

BACKGROUND & AIMS: Pancreatic thread proteins (PTPs) are acinar cell products and members of the regenerating gene (reg) family. reg expression increases during islet regeneration, is depressed during aging-related islet dysfunction, and may be important in beta-cell growth and maintenance. The aim of this study was to examine the genetic expression of reg in pancreatic-derived cells in vitro and the mitogenic effect of PTP/reg protein on these cells. METHODS: reg gene expression was measured by Northern analysis in three rat pancreatic cell lines: ARIP (ductal), AR42J (acinar), and RIN (beta-cell). PTP/reg protein was isolated from bovine and human pancreas. Cells were cultured with PTP/reg for 72 hours, and thymidine incorporation was measured. RESULTS: reg messenger RNA was detected in AR42J but not in ARIP or RIN. PTP/reg protein was mitogenic to RIN and ARIP in a dose-related fashion but not to AR42J. It was not mitogenic to cultured mature rat islets. CONCLUSIONS: reg messenger RNA is expressed in acinar but not in beta-cell or ductal pancreatic cell lines. PTP/reg protein was mitogenic to both beta-cell and ductal cell lines but not to mature, nondividing islets. This supports the hypothesis that PTP/reg protein is an acinar cell-derived mediator of beta-cell growth and may be involved in modulating the duct-to-islet axis.

Animals↗
PubMed1996

Differential expression of reg-I and reg-II genes during aging in the normal mouse.

A cDNA termed reg (for regenerating gene) has been isolated from a rat pancreatic DNA library. Reg expression has been shown to correlate with changes in beta cell mass and function. This finding has been recently challenged by studies showing a non-beta-cell-dependent regulation of reg expression. All studies to date, however, have neglected the fact that two nonallelic reg genes (reg-I and reg-II) exist in several species. In studying the regulation of each individual copy gene, we investigated reg-I and -II gene expression in a naturally occurring modification of beta-cell physiology: normal aging. RNA was isolated from individual pancreata of 1-, 3-, 9-, 20-, and 30-month-old C57BL/6J mice (n > or = 3 per group) and subjected to slot-blot analysis using homologous probes for reg-I, reg-II, insulin, and elastase-I. A progressive age-dependent decrease in total reg mRNA levels (reg-I and -II) was detected. At 30 months of age, total reg mRNA levels were approximately 45% of the level detected in 1-month-old mice (p = .01). This paralleled the decrease in insulin mRNA levels (p = .01), which fell below 50%; by contrast, mRNA levels for elastase-I increased with age (p = .05). Analysis of RNA isolated from purified islets did not reveal any mRNA for reg, suggesting that in the normal mouse, reg is primarily a product of the exocrine pancreas. Reg mRNA were detectable in RNA extracts from stomach, duodenum, and small intestine. By hybridization of total pancreatic RNA with oligonucleotide probes which specifically recognize reg-I or reg-II sequences, we show that reg-I mRNA levels declined with age (p = .001) while reg-II mRNA levels remained unchanged. These data demonstrate that in mouse pancreas the two nonallelic reg genes are differentially expressed during aging and that the decrease in reg-I mRNA levels parallels the decrease in insulin gene expression. Differential regulation of reg-I and reg-II genes may explain the presence of conflicting data in the current literature.

Aging↗
PubMed1996

Altered insulin receptor messenger ribonucleic acid splicing in liver is associated with deterioration of glucose tolerance in the spontaneously obese and diabetic rhesus monkey: analysis of controversy between monkey and human studies.

There are two insulin receptor (IR) isoforms (designated type A and type B), derived from alternative splicing of exon 11 of the IR gene. Recently, we reported (Huang Z., Bodkin N.L., Ortmeyer H.K., Hansen B.C., Shuldiner A. R., 1994, J Clin Invest, 94:1289-1296) that an increase in the exon 11- (i.e. lacking exon 11) (type A) IR messenger RNA (mRNA) variant in muscle is associated with hyperinsulinemia, an early risk factor for noninsulin-dependent diabetes mellitus (NIDDM), in the spontaneously obese, diabetic rhesus monkey. To explore further the role of IR mRNA splicing in insulin resistance of NIDDM, we studied liver, another target organ that is resistant to insulin action in NIDDM. The relative amounts of the two IR mRNA-splicing variants in liver were quantitated by RT-PCR in normal, prediabetic, and diabetic (NIDDM) monkeys. The percentage of the exon 11- mRNA variant in liver (n = 24) was significantly correlated with fasting plasma glucose (r = 0.55, P < 0.01) and intravenous glucose disappearance rate (r = -0.45, P < 0.05). The exon 11- mRNA variant was increased significantly from 29.8 +/- 1.6% in monkeys with normal fasting glucose to 39.2 +/- 2.9% in monkeys with elevated fasting glucose (P < 0.01). These studies provide the first direct evidence in vivo that the relative expression of the two IR mRNA-splicing variants is altered in liver and suggest that increased expression of the exon 11- IR isoform may contribute to hepatic insulin resistance and NIDDM or may compensate for some yet unidentified defect.

Alternative Splicing↗
PubMed1996

Regenerating (reg) and insulin genes are expressed in prepancreatic mouse embryos.

The pancreatic regenerating (reg) gene is proposed to be involved in pancreatic beta-cell growth. Up- or down-regulation of reg gene expression has been shown to parallel variations in beta-cell mass and function in the adult pancreas. In several species at least two nonallelic reg genes have been identified. In this study we investigated the expression of each individual reg gene (reg-I and reg-II) during embryogenesis in the mouse. Single mouse embryos were harvested at 8.5, 9, 10, and 12 days of development, homogenized and subjected individually to reverse transcription (RT)-PCR, with a single primer pair to amplify both reg-I and -II mRNAs. Southern blot analysis of the RT-PCR products revealed the presence of reg mRNA at day 9 of embryogenesis, just before the beginning of pancreatic organogenesis. Slot-blot analysis with internal oligonucleotide probes that specifically recognize reg-I or -II sequences demonstrated that only reg-I mRNA was present in day 9 and day 10 prepancreatic embryos. Reg-II mRNA was not detected until day 12, a stage corresponding to late organogenesis. RT-PCR for insulin mRNA from the same samples used for the amplification of reg mRNA showed that the earliest insulin expression occurred at day 8.5, and coincided with the onset of reg-I expression. Hybridization with gene-specific oligonucleotide probes revealed that only insulin-II mRNA was detectable at this time. Insulin-I mRNA was not detectable until day 12 and coincided with early reg-II expression. These results suggest that the two nonallelic reg genes and the two insulin genes are expressed differentially during early embryogenesis. Differential expression of reg-I and -II suggests that they may be induced by different and independent stimuli and have distinct functions.

Animals↗
PubMed1996

Outcome following colon surgery in the octagenarian.

The results of colon surgery in all individuals aged 80 years or greater at one teaching institution during the 1987-1993 time period were reviewed. Sixty patients, ranging in age from 80 to 92 years, underwent 41 elective operations and 21 emergency procedures. Emergency procedures resulted in death or a major complication in over one-half of patients, and only six were ultimately able to return home. Conversely, elective procedures were relatively well tolerated, and 31 of 37 survivors returned immediately to independent living (P = 0.006). Mortality was 33.3 per cent in emergency cases versus 9.8 per cent in elective operations (P < 0.03). The occurrence of a postoperative complication increased the length of stay by an average of 12 days. These data suggest that elective colon surgery in the elderly produces results little different from the population at large. Conversely, emergency operations are associated with a high morbidity and mortality rate. Age alone should not be a determining factor in who undergoes an elective colon operation. Greater efforts should be made to screen elderly individuals to limit emergency surgery.

Age Factors↗