Translation regulatory factor RBM3 is a proto-oncogene that prevents mitotic catastrophe.

RNA-binding proteins play a key role in post-transcriptional regulation of mRNA stability and translation. We have identified that RBM3, a translation regulatory protein, is significantly upregulated in human tumors, including a stage-dependent increase in colorectal tumors. Forced RBM3 overexpression in NIH3T3 mouse fibroblasts and SW480 human colon epithelial cells increases cell proliferation and development of compact multicellular spheroids in soft agar suggesting the ability to induce anchorage-independent growth. In contrast, downregulating RBM3 in HCT116 colon cancer cells with specific siRNA decreases cell growth in culture, which was partially overcome when treated with prostaglandin E(2), a product of cyclooxygenase (COX)-2 enzyme activity. Knockdown also resulted in the growth arrest of tumor xenografts. We have also identified that RBM3 knockdown increases caspase-mediated apoptosis coupled with nuclear cyclin B1, and phosphorylated Cdc25c, Chk1 and Chk2 kinases, implying that under conditions of RBM3 downregulation, cells undergo mitotic catastrophe. RBM3 enhances COX-2, IL-8 and VEGF mRNA stability and translation. Conversely, RBM3 knockdown results in loss in the translation of these transcripts. These data demonstrate that the RNA stabilizing and translation regulatory protein RBM3 is a novel proto-oncogene that induces transformation when overexpressed and is essential for cells to progress through mitosis.

Systemic bone loss and induction of coronary vessel disease in a rat model of chronic inflammation.

Clinically, osteopenia or low bone mass has been observed in a variety of chronic inflammatory diseases, and elevated proinflammatory mediators have implicated this process. The purpose of this study was to develop an in vivo model of bone loss induced by chronic systemic inflammation. Time-release pellets designed to deliver one of three doses of LPS: Low (3.3 microg/day), High (33.3 microg/day), or Placebo over 90 days, were implanted subcutaneously in 3-month-old male Sprague-Dawley rats (n = 8/group). Neutrophil counts, indicative of ongoing inflammation, were elevated (P < 0.05) in both LPS groups at 30 days post-implant and remained significantly elevated in the High dose throughout the 90-day study period. At the end of the study, bone loss occurred in the femur as indicated by decreased bone mineral density (BMD) in both LPS-treated groups, but vertebral BMD was reduced in the High dose animals only. Microcomputed tomography revealed that trabecular bone volume (BV/TV) of the proximal tibial metaphysis tended to be reduced in the High dose LPS group. Deleterious effects on trabecular number (TbN) and trabecular separation (TbSp) were observed in both LPS-treated groups, but only the High dose group reached statistical significance. These alterations in trabecular microarchitecture resulted in compromised biomechanical properties. No changes in cortical thickness, porosity, or area of the tibia midshaft were evident at either dose of LPS. Up-regulation of the proinflammatory mediators, cyclooxygenase (COX)-2, interleukin (IL)-1, and tumor necrosis factor (TNF)-alpha was demonstrated in the metaphyseal region where the deleterious effects of LPS were observed. In addition to these alterations in bone, trichrome staining indicated changes in the coronary arterioles, consistent with vascular disease. Utilization of a LPS time-release pellet appears to provide an in vivo model of chronic inflammation-induced bone loss and a potentially novel system to study concurrent development of osteopenia and vascular disease.

Assessment of the safety of recombinant tissue factor pathway inhibitor in patients with severe sepsis: a multicenter, randomized, placebo-controlled, single-blind, dose escalation study.

OBJECTIVE: To identify a safe and potentially effective recombinant tissue factor pathway inhibitor (rTFPI) dose for further clinical evaluation in patients with severe sepsis. DESIGN: Prospective, randomized, single-blind, placebo-controlled, dose escalation, multicenter, multinational phase II clinical trial. SETTING: Thirty-eight intensive care units in the United States and Europe. PATIENTS: Two hundred and ten subjects with severe sepsis who received standard supportive care and antimicrobial therapy. INTERVENTIONS: Subjects received a continuous intravenous infusion of placebo or rTFPI at 0.025 or 0.05 mg/kg/hr for 4 days (96 hrs). MEASUREMENTS AND MAIN RESULTS: There were no significant imbalances in demographics, severity of illness, or source of infection in patients randomized to placebo or either dose of rTFPI. A 20% relative reduction in 28-day all-cause mortality was observed when all rTFPI-treated patients were compared with all placebo patients. An improvement in pulmonary organ dysfunction score and in a composite intensive care unit score (pulmonary, cardiovascular, and coagulation) were also noted in the rTFPI-treated patients. Logistic regression modeling indicated a substantial treatment by baseline laboratory international normalized ratio (INR) interaction effect when only treatment and INR were in the model (p =.037) and when baseline Acute Physiology and Chronic Health Evaluation (APACHE II) and log10 interleukin 6 were adjusted for (p =.026). This interaction effect indicates that higher baseline INR is associated with a more pronounced beneficial rTFPI effect. There was no increase in mortality in subjects treated with either dose of rTFPI compared with placebo. Biological activity, as detected by a statistically significant reduction in thrombin-antithrombin complexes (TATc), was noted in the all rTFPI-treated patients compared with those receiving placebo. There were no major imbalances across all treatment groups with respect to safety. The frequency of adverse events (AEs) and severe adverse events (SAEs) was similar among the treatment groups, with a slight increase in SAEs and SAEs involving bleeding in the 0.05 mg/kg/hr rTFPI group. The overall incidence of AEs involving bleeding was 28% of patients in the all placebo group and 23% of patients in the all rTFPI-treated group; a slight but statistically insignificant increase in incidence of SAEs involving bleeding was observed in the all rTFPI group (9%) as compared with the all placebo group (6%; p =.39). CONCLUSIONS: Although the trial was not powered to show efficacy, a trend toward reduction in 28-day all-cause mortality was observed in the all rTFPI group compared with all placebo. This study demonstrates that rTFPI doses of 0.025 and 0.05 mg/kg/hr could be safely administered to severe sepsis patients. Additionally, rTFPI demonstrated bioactivity, as shown by reduction in TATc complexes and interleukin-6 levels. These findings warrant further evaluation of rTFPI in an adequately powered, placebo controlled, randomized trial for the treatment of severe sepsis.

Past, present, and future of pancreatic surgery.

The advances in the surgical treatment of pancreatic disease in the 20th century were built on careful anatomic and physiologic studies dating to the early 1800s. Operations for neoplastic diseases developed in the 1930s by Whipple, Trimble, and others allowed pancreatic malignancies to be removed with ever increasing safety. Endocrine tumors of the pancreas were described and treated surgically. Patients with pancreatitis now have a number of surgical alternatives available for their individual circumstances. The future of surgery for pancreatic disease lies in the results of the human genome project and the fields of genomics and proteomics that resulted. The rapidity with which knowledge of gene expression is advancing owing to new technologies such as the microarray biochip is amazing. The future of pancreatic surgery is bright.

Comparison of postoperative outcomes in ulcerative colitis and familial polyposis patients after ileoanal pouch operations.

BACKGROUND: Pouchitis is a poorly understood inflammatory condition that occurs in the ileal pouches of patients who have undergone the ileal-pouch anal anastomosis after restorative proctocolectomy. This postoperative condition is much more common in patients with ulcerative colitis (UC) than familial adenomatous polyposis (FAP) colitis. It has been suggested that, owing to pouchitis, UC patients do not attain the same quality of life that FAP patients do after the ileal-pouch anal anastomosis operation. We hypothesized that health-related quality of life does not differ between FAP and UC patients. METHODS: We analyzed the postoperative morbidity and gastrointestinal function in 110 consecutive patients having undergone the ileal-pouch anal anastomosis for either UC or FAP at OU Medical Center from 1983 to 2000 by retrospective record review. Health-related quality of life was assessed in 83 patients using the Short Inflammatory Bowel Disease Questionnaire (SIBDQ) and the Medical Outcome Study Short-Form 36 (SF-36) questionnaire. RESULTS: With the exception of pouchitis, there was no difference in perioperative outcome, morbidity, or functional status between UC and FAP patients. The SIBDQ and SF-36 revealed no statistically significant difference between FAP and UC patients. CONCLUSIONS: As expected, UC patients are more likely to develop pouchitis. Despite this, our data reveal that both patient groups enjoy a similarly good functional status and quality of life.

Antibiotic-resistant organism infection.

Bacteria possess a remarkable number of ways to become resistant to antibiotics. Antibiotic resistance has become a major problem in the treatment of Gram-positive infections. Resistance to methicillin and vancomycin in staphylococci and enterococci has resulted in organisms that are resistant to all known antibiotics. Although it is important to continue to search for newer and more effective antibiotics, it is imperative that we develop a surgical mindset of appropriate antibiotic stewardship. The use of single-dose prophylactic regimens, using narrow-spectrum agents when possible for therapeutic indications, limiting the duration of therapeutic agents appropriately, avoiding the use of vancomycin except when necessary, and adhering to strict infection control measures are all steps that will limit the spread and development of resistant organisms.

Effect of PGG-glucan on the rate of serious postoperative infection or death observed after high-risk gastrointestinal operations. Betafectin Gastrointestinal Study Group.

BACKGROUND: Postoperative infections remain common after high-risk gastrointestinal procedures. PGG-glucan (Betafectin; Alpha Beta Technology Inc, Worcester, Mass), derived from yeast cell walls, promotes phagocytosis and intracellular killing of bacterial pathogens by leukocytes, prevents infection in an animal model of wound infection, and acts synergistically with antibiotics to reduce mortality in rat peritonitis. HYPOTHESIS: We hypothesized that infectious complications in these patients might be reduced by the administration of a nonspecific immune-enhancing agent. DESIGN: Multicenter, prospective, randomized, double-blind, placebo-controlled trial of 1249 patients prospectively stratified into colorectal or noncolorectal strata. SETTING: Thirty-nine medical centers throughout the United States. PATIENTS: Aged 18 years or older, scheduled for gastrointestinal procedure lasting 2 to 8 hours, with 2 or more defined risk factors. INTERVENTIONS: PGG-glucan, 0.5 mg/kg or 1.0 mg/kg, or placebo once preoperatively and 3 times postoperatively. All patients received standardized antibiotic prophylaxis. MAIN OUTCOME MEASURES: Serious infection or death within 30 days. RESULTS: All randomized patients revealed no difference in serious infections and deaths in the treated groups compared with placebo groups (15% vs 14%, P>.90). In the prospectively defined noncolorectal stratum (n = 391), PGG-glucan administration was associated with a statistically significant relative reduction (39%) in serious infections and death (placebo, 46 [36%] of 129 vs either PGG-glucan group, 29 [21%] of 132 and 28 [22%] of 130, P<.02). PGG-glucan reduced postoperative infection or death in malnourished patients having noncolorectal procedures (31 [44%] of 70, placebo group; 16 [24%] of 68, 0.5-mg/kg PGG-glucan group; 12 [17%] of 72, 1.0-mg/kg PGG-glucan group; P<.001). Study drug was stopped owing to adverse effects more frequently for patients receiving PGG-glucan than placebo (2%, 4%, and 7% for the placebo group, 0.5-mg/kg PGG-glucan group, and 1.0-mg/kg PGG-glucan group, respectively, P<.003). CONCLUSION: Perioperative administration of PGG-glucan reduced serious postoperative infections or death by 39% after high-risk noncolorectal operations.

Expression of the cyclin-dependent kinase inhibitor p21(WAF1/CIP1) and p53 tumor suppressor in dysplastic progression and adenocarcinoma in Barrett esophagus.

BACKGROUND: Barrett esophagus predisposes individuals to esophageal carcinoma, which develops from intermediate stages of tissue dysplasia primarily in the vicinity of the gastroesophageal junction. Understanding the cellular and molecular events in the progression of Barrett esophagus to adenocarcinoma may contribute to its early diagnosis and treatment. Mutation and overexpression of the tumor suppressor p53 have previously been observed in Barrett high grade dysplasia and adenocarcinoma. The expression of the cyclin-dependent kinase (CdK) inhibitor p21 can be up-regulated by p53, resulting in the down-regulation of cell division at the G(1)/S-phase transition. The current study examined the correlation between the expression of p21 and p53 by quantifying their levels during the progression of dysplasia and adenocarcinoma in Barrett esophageal tissues. METHODS: Barrett esophageal tissue samples that were negative or indefinite for dysplasia, contained dysplasia, and contained adenocarcinoma were examined by immunohistochemistry. Paraffin embedded sections of lining and glandular epithelia were adsorbed with primary murine antibodies against human p21 or p53 followed by horseradish peroxidase secondary antibody. An immunoreactivity score for each primary antibody and section was obtained by multiplying a staining intensity factor by the percent of positively stained cells. RESULTS: Nuclear p21 expression was detectable immunohistochemically in Barrett esophagus that was negative for dysplasia, but it was significantly elevated (P

Colorectal cancer: genetics and screening.

Colorectal cancer is a common disease in the Western world. Most, if not all, colorectal cancers develop from previously benign adenomas. There are a number of genetic abnormalities including mutations in oncogenes and tumor suppressor genes which either present as a germline, or acquired defects lead to the development of colorectal cancer. Two well-defined hereditary colorectal cancer syndromes exist, hereditary nonpolyposis colorectal cancer syndrome and familial adenomatous polyposis coli, for which genetic testing is possible and advised. Guidelines for screening for colorectal cancer in average, moderate, and high risk patients are available from the American Cancer Society and were updated in 1997. The American Society of Clinical Oncology has published guidelines for genetic testing in a variety of cancers including colorectal cancer.

Pancreatic adenocarcinoma: a review for primary care physicians.

Adenocarcinoma of the pancreas is becoming an increasingly common disease. The differential diagnosis of pancreatic adenocarcinoma is that of obstructive jaundice. Suspicious findings on history and physical examination can be confirmed with appropriate laboratory and radiologic testing. Approximately 20% of patients with small lesions and no metastatic disease may be cured with resection. The operative mortality and morbidity for major pancreatic resections is now sufficiently low to warrant a more aggressive approach to these patients.

Repair of parastomal hernias using polypropylene mesh.

Parastomal hernias are a common complication of ostomy construction. We have developed a method of repair that uses two strips of polypropylene prosthetic mesh through a midline incision. The medical records of 19 patients who underwent parastomal hernia repair were retrospectively reviewed. All nine patients operated on for this condition by the senior author (R.G.P.) (group 1) underwent repairs with this technique. All ten patients operated on by other surgeons in our center (group 2) underwent repairs in which the stoma was moved, the fascia was directly repaired through a parastomal incision, or the fascia was repaired via a midline incision. No patients in group 1 had recurrences while five patients in group 2 had recurrences. Neither group developed strictures or stomal prolapse. Our method of repair is technically easy and has excellent results. It is especially suitable in very large hernias in which incisional hernia is likely in the original stoma site if the stoma is moved.

Early detection of colorectal cancer by quantitative fluorescence image analysis of exfoliated cells.

Early-stage colorectal cancer is potentially curable. In the present study, we applied quantitative fluorescence image analysis (QFIA) cytology to the detection of experimental colorectal cancer in a rodent model. QFIA cytology combines visual cytologic examination with quantitation of DNA content in single exfoliated cells. Cancer was induced by treating 110 rats with subcutaneous 1,2-dimethylhydrazine. Sequential colon washes were obtained weekly from each animal for 20 weeks. Control animals were treated identically except for the administration of carcinogen. Cells that were cytologically abnormal or had increased DNA content were found starting in the second week. By the eighth week, roughly 50 percent of animals had positive results, and this level remained approximately constant for the duration of the study. Tissue pathologic results were normal during weeks 1 to 7. Dysplasias became common during weeks 8 to 15 whereas most cancers appeared during weeks 16 to 21. These results indicate that QFIA cytology is a highly sensitive method for detecting even preneoplastic changes resulting from carcinogen administration and may prove useful in detecting human colorectal cancer.

Factors predicting failure of medical therapy for gastric ulcers.

We reviewed the charts of all patients admitted with a diagnosis of gastric ulcer from January 1970 to December 1980. Multiple risk factors were recorded in patients receiving medical treatment and compared in those patients successfully treated medically versus those requiring operation after a failed course of medical treatment. One hundred patients were treated medically without surgical intervention, and 34 patients underwent operation after medical therapy failed. Significant risk factors in patients requiring operative therapy included smoking (p = 0.03), multiple trauma and sepsis (p = 0.02), large ulcers (p = 0.03), and multiple ulcers (p = 0.02). We have identified a set of factors associated with a high risk of failure of medical therapy. Patients with any of these risk factors may be treated most effectively by a limited trial of medical therapy with close follow-up. If their ulcer disease does not respond readily to standard medical therapy, they should be considered for early elective surgery.

Adenocarcinoma of the colon in adolescents.

A retrospective study was carried out in nine children between the ages of 10 and 20 years with adenocarcinoma of the colon. No family history, significant medical history, or predisposing factors were identified, except for Turcot's syndrome in one child. Common presenting signs and symptoms were vague abdominal pain, nausea and vomiting, weight loss, change in bowel habits, and guaiac-positive stools. Five of the patients' diagnoses were delayed for an average of 11.6 months, the majority of whom had Dukes' D disease. Their median survival was 4 months compared with 24 months in the four patients diagnosed early. As with adults, the mainstay of therapy is operation. Our data indicate that an increased awareness and consideration of colon cancer in children will result in earlier diagnosis, a more favorable disease stage, and prolonged survival.

Equilibrium binding of Hoechst 33258 and Hoechst 33342 fluorochromes with rat colorectal cells.

We examined the biophysical characteristics of the interaction of Hoechst 33258 and 33342 dyes with normal rat colorectal cells as functions of fixation and solution composition. Classical dye-binding techniques were used to investigate the stoichiometry and binding constants with whole cells, and quantitative fluorescence image analysis was used to specifically study nuclear dye binding in intact cells. In aqueous solution, H-33258 dye bound cooperatively with intact cells, with a binding constant of between 3-4 x 10(5). In ethanolic solution, binding appeared less cooperative, although Scatchard analysis could not be used. The binding constant was slightly lower (2 x 10(5)), but the total number of cell binding sites was decreased by a factor of 5, reflecting a great decrease in cytoplasmic sites. QFIA studies identified conditions optimal for DNA quantitation under which the fluorescence signal was independent of dye or cell concentration. The proportionality between absolute nuclear fluorescence intensity and DNA content was established, and the upper limit of DNA content of normal colorectal cells was also determined.

Treatment of esophageal cancer: the continued need for surgical resection.

We conclude that preoperative radiotherapy and chemotherapy is very likely to improve the length of survival in patients with squamous cell carcinoma undergoing surgical resection and decreases the incidence of symptomatic local recurrence requiring additional palliative treatment. A favorable tumor response to adjuvant therapy, however, did not significantly improve the survival curves in our study, does not guarantee complete tumor regression, and should not be cited as a basis for elimination of esophagectomy in treatment protocols. Adenocarcinoma treated with surgical resection has a better prognosis than similarly treated squamous cell carcinoma, but has a poorer prognosis than squamous cell carcinoma treated with preoperative adjuvant therapy and surgical resection.

The fluosol-perfused isolated canine pancreas: a model for the study of blood component effects in acute pancreatitis.

UNLABELLED: Blood components have been implicated as factors which modulate organ injury in acute pancreatitis. To isolate these effects we compared a standardized isolated, blood-perfused, canine pancreas model as described by Herman-Taylor and modified by Cameron with a model using fluosol, a fluorocarbon, as the perfusate. Pancreatitis was induced using partial pancreatic duct obstruction with secretin induced hypersecretion. Twenty-four dogs were randomized into four groups; (1) blood-perfused control (BPC), (2) fluosol-perfused control (FPC), (3) blood-perfused injury (BPI), (4) fluosol-perfused injury (FPI). All glands were observed for 4 h for mean arterial pressure (MAP), weight gain, gross appearance, and venous amylase. Stability was monitored with blood gases and glucose. Specimens were taken at 4 h for light and electron microscopy. RESULTS: BPI preps had a significantly higher vascular resistance at 3 and 4 h compared to FPI preps (3.85 vs 3.26 PRU and 4.8 vs 3.9 PRU, respectively) (P less than 0.002). Edema formation (3+ vs 1+) and venous amylase (18,543 vs 1961) (P less than 0.001) were greater in BPI than FPI preps. Light and electron microscopy confirmed injury but could not quantify it between injury groups. SUMMARY: Hypersecretion and partial ductal obstruction causes a more severe injury in the blood than the fluosol perfused preparation. Changes in the peripheral resistance seen in the blood perfused model lend support to the theory that the primary injury in this model is at the capillary level and is modulated by a blood component.