Identification of FAP locus genes from chromosome 5q21.

Recent studies suggest that one or more genes on chromosome 5q21 are important for the development of colorectal cancers, particularly those associated with familial adenomatous polyposis (FAP). To facilitate the identification of genes from this locus, a portion of the region that is tightly linked to FAP was cloned. Six contiguous stretches of sequence (contigs) containing approximately 5.5 Mb of DNA were isolated. Subclones from these contigs were used to identify and position six genes, all of which were expressed in normal colonic mucosa. Two of these genes (APC and MCC) are likely to contribute to colorectal tumorigenesis. The MCC gene had previously been identified by virtue of its mutation in human colorectal tumors. The APC gene was identified in a contig initiated from the MCC gene and was found to encode an unusually large protein. These two closely spaced genes encode proteins predicted to contain coiled-coil regions. Both genes were also expressed in a wide variety of tissues. Further studies of MCC and APC and their potential interaction should prove useful for understanding colorectal neoplasia.

Identification of a gene located at chromosome 5q21 that is mutated in colorectal cancers.

Recent studies have suggested the existence of a tumor suppressor gene located at chromosome region 5q21. DNA probes from this region were used to study a panel of sporadic colorectal carcinomas. One of these probes, cosmid 5.71, detected a somatically rearranged restriction fragment in the DNA from a single tumor. Further analysis of the 5.71 cosmid revealed two regions that were highly conserved in rodent DNA. These sequences were used to identify a gene, MCC (mutated in colorectal cancer), which encodes an 829-amino acid protein with a short region of similarity to the G protein-coupled m3 muscarinic acetylcholine receptor. The rearrangement in the tumor disrupted the coding region of the MCC gene. Moreover, two colorectal tumors were found with somatically acquired point mutations in MCC that resulted in amino acid substitutions. MCC is thus a candidate for the putative colorectal tumor suppressor gene located at 5q21. Further studies will be required to determine whether the gene is mutated in other sporadic tumors or in the germ line of patients with an inherited predisposition to colonic tumorigenesis.

Progression of colorectal cancer is associated with multiple tumor suppressor gene defects but inhibition of tumorigenicity is accomplished by correction of any single defect via chromosome transfer.

Carcinogenesis is a multistage process that has been characterized both by the activation of cellular oncogenes and by the loss of function of tumor suppressor genes. Colorectal cancer has been associated with the activation of ras oncogenes and with the deletion of multiple chromosomal regions including chromosomes 5q, 17p, and 18q. Such chromosome loss is often suggestive of the deletion or loss of function of tumor suppressor genes. The candidate tumor suppressor genes from these regions are, respectively, MCC and/or APC, p53, and DCC. In order to further our understanding of the molecular and genetic mechanisms involved in tumor progression and, thereby, of normal cell growth, it is important to determine whether defects in one or more of these loci contribute functionally in the progression to malignancy in colorectal cancer and whether correction of any of these defects restores normal growth control in vitro and in vivo. To address this question, we have utilized the technique of microcell-mediated chromosome transfer to introduce normal human chromosomes 5, 17, and 18 individually into recipient colorectal cancer cells. Additionally, chromosome 15 was introduced into SW480 cells as an irrelevant control chromosome. While the introduction of chromosome 17 into the tumorigenic colorectal cell line SW480 yielded no viable clones, cell lines were established after the introduction of chromosomes 15, 5, and 18. Hybrids containing chromosome 18 are morphologically similar to the parental line, whereas those containing chromosome 5 are morphologically distinct from the parental cell line, being small, polygonal, and tightly packed. SW480-chromosome 5 hybrids are strongly suppressed for tumorigenicity, while SW480-chromosome 18 hybrids produce slowly growing tumors in some of the animals injected. Hybrids containing the introduced chromosome 18 but was significantly reduced in several of the tumor reconstitute cell lines. Introduction of chromosome 5 had little to no effect on responsiveness, whereas transfer ot chromosome 18 restored responsiveness to some degree. Our findings indicate that while multiple defects in tumor suppressor genes seem to be required for progression to the malignant state in colorectal cancer, correction of only a single defect can have significant effects in vivo and/or in vitro.

Wild-type but not mutant APC associates with the microtubule cytoskeleton.

The adenomatous polyposis coli protein (APC) is mutated in familial adenomatous polyposis patients as well as in sporadic colorectal tumors. In an attempt to further understand the function of APC, the subcellular localization of APC was examined. Wild-type and mutant forms of APC were expressed in mammalian cells and protein detected by immunofluorescence using monoclonal and polyclonal antibodies. Staining of wildtype APC protein revealed a filamentous network which extended throughout the cytoplasm and colocalized with microtubules. In striking contrast, mutant APC protein gave a diffuse cytoplasmic staining pattern. Treatment with a microtubule depolymerizing agent, nocodazole, caused APC as well as tubulin to become diffusely cytoplasmic. In addition, immunoperoxidase staining of transfected APC protein followed by transmission electron microscopy revealed staining of microtubules. These results suggest that wild-type but not mutant APC protein may be associated with the microtubule cytoskeleton.

Inactivation of both APC alleles in human and mouse tumors.

Germline mutations of the adenomatous polyposis coli (APC) gene lead to multiple intestinal tumors in familial adenomatous polyposis patients and in multiple intestinal neoplasia (Min) mice. Current models predict that inactivation of the remaining normal allele of a tumor suppressor gene is rate limiting for tumor formation, but this has been difficult to prove. While examination of colorectal adenomas from familial adenomatous polyposis patients identified somatic inactivating mutations of the second allele in the majority of tumors (19 of 24), the absolute requirement for an early inactivating event could not be demonstrated. In contrast, inactivation of the remaining allele of the murine APC (Apc) could be demonstrated in 100% (30 of 30) of tumors from Min mice. Moreover, inactivation was observed in the earliest recognizable phase of tumors, including some lesions containing as few as two dysplastic crypts. These results suggest that the mutation of the second APC allele is an early event in Min and familial adenomatous polyposis tumorigenesis, supporting Knudson's hypothesis.

Combination dihydroergotamine mesylate and heparin sodium with lidocaine HCl. Pharmacokinetics, mechanism of action, clinical efficacy, and adverse effects.

Dihydroergotamine(DHE)-heparin combination offers a unique treatment modality for the prevention of deep vein thrombosis. The combination appears to affect all 3 limbs of Virchow's triad: hypercoagulability, venous stasis, and endothelial damage. In most efficacy studies, data indicated that the combination of DHE 0.5 mg and heparin 5000 IU was superior to low-dose heparin alone. Even when the efficacy of DHE-heparin was the same as that of heparin alone, the use of the combination allowed for a decrease in the heparin dose required.

Geriatric trauma.

Elderly individuals not only live longer but are also more active than in the past. Accompanying this increase in activity is the number of older trauma victims. The effect of aging on response to injury is reviewed, and the initial evaluation and treatment of geriatric trauma are delineated in this article. Specific injuries of note include head and chest wall injury; pulmonary and cardiac contusion; abdominal trauma; and aortic, spinal, and musculoskeletal injury.

Documentation of clinical and cost-saving pharmacy interventions in the emergency room.

Hospital pharmacists have shown clearly their benefit in patient care by intervening to improve the drug use process. In this era of cost containment, hospital administrators are likely to fund only those programs that clearly improve patient care or reduce costs. To demonstrate the impact on a hospital budget and to justify a position or service, documentation of improvement and generation of a cost-savings report is essential. This article discusses the types and methods of clinical and cost-saving interventions that are made in a busy inner city university trauma center's emergency department, and the methods by which our data are collected.

Anaphylactic reaction due to zomepirac.

Zomepirac sodium is a nonsteroidal antiinflammatory drug with potent analgesic properties. Since its introduction in October 1980, this prostaglandin inhibitor has been fairly well tolerated, and little toxicity has been associated with its use. In fact, it was not until April 1981 that the first serious reaction of anaphylaxis was described. Since then six additional cases have been reported to the literature. The manufacturer voluntarily withdrew the agent in March 1983 from the market pending further investigation of these reactions. We describe the case of an additional anaphylactic reaction to zomepirac and review the literature.

Mineral Precipitation and Trace Oxyanion Behavior during Evaporation of Saline Waters.

Evaporation basins used for the disposal of agricultural drainage waters in central California contain elevated trace element levels that pose hazards to groundwater quality and wildlife visiting the ponds. A study was conducted to evaluate the solution dynamics of mineral-forming elements and trace oxyanions (U, Mo, and V) during evaporation of saline waters whose chemical compositions evolve according to two distinct chemical divides, and to characterize the evaporite minerals formed from the complete evaporation of these waters. The alkali and alkaline earth metals exhibited nonconservative behavior, forming evaporite minerals such as bloedite, calcite, aragonite, gypsum, halite, thenardite, and trona. Molybdenum behaved nonconservatively, while V exhibited conservative behavior that did not differ whether V was initially added as V(IV) or V(V). Uranium displayed conservative behavior under conditions of low U concentrations and high alkalinities. Nonconservative behavior was observed for U, however, under higher U concentrations and low alkalinities. We conclude that V and U in waters with alkalinities >10 mmolc L-1 will not accumulate in evaporation pond minerals. In ponds with low alkalinity, U will partition to a solid mineral phase.

Lactic acidosis and acute ethanol intoxication.

Ethanol intoxication has been widely reported as a cause of lactic acidosis. To determine the frequency and severity of ethanol-induced lactic acidosis, patients who presented to an emergency department with a clinical diagnosis of acute ethanol intoxication and a serum ethanol concentration of at least 100 mg/dL were studied. Arterial blood was sampled for lactate and blood gas determinations. A total of 60 patients (mean age, 41 years) were studied. Twenty-two patients sustained minor trauma. Ethanol concentrations ranged from 100 to 667 mg/dL (mean, 287 mg/dL). Lactate concentrations were abnormal (> 2.4 mmol/L) in seven patients (11.7%). In all cases, blood lactate was less than 5 mmol/L. Of the patients with elevated lactate, other potential causes for lactic acidosis, including hypoxia, seizures, and hypoperfusion, were also present. Only one case with elevated blood lactate concentration had associated acidemia. Significant elevations of blood lactate are uncommon in acute ethanol intoxication. In patients with ethanol intoxication who are found to have lactic acidosis, other etiologies for the elevated lactate level should be considered.

Identification of a phorbol ester-repressible v-src-inducible gene.

Chicken embryo fibroblasts (CEF) infected with a temperature-sensitive Rous sarcoma virus (RSV) mutant, tsNY72-4, express a set of pp60v-src-induced RNAs soon after shift to the permissive temperature. By subtractive and differential screening, we have cloned 12 of these sequences, 2 of which were c-fos and krox-24. Serum induced all the v-src-inducible genes tested, suggesting that these genes serve roles in normal cell division and are not specific to transformation per se. Significantly, however, v-src produced prolonged, and in some cases kinetically complex, patterns of induction compared to serum. For most of the clones, phorbol 12-tetradecanoate 13-acetate (TPA) induced mRNAs with kinetics similar to that of serum. However, one clone (CEF-4) was expressed in a biphasic manner. Another (CEF-10) was repressed by TPA at 1 hr, after which this mRNA was permanently induced. The pattern of repression-induction of CEF-10 mRNA is the inverse of protein kinase C (PKC) activity in the cell, suggesting that PKC actively represses this gene. In vivo expression of CEF-10 mRNA is restricted predominantly to the lung. A full-length CEF-10 cDNA encodes a 41-kDa protein that has an amino-terminal signal peptide for secretion, contains a markedly high number of cysteine residues, and shows no sequence similarity to known proteins.

Menarche: target age for reinforcing sex education for adolescents.

Teenage sexual activity has led to an increasing prevalence of pregnancy and sexually transmitted disease in this age group. Attempts at prevention should begin with education. This study compares the ages of menarche with ages of first intercourse among adolescent females in 1971 and 1976, stratified by ethnic group and residence (urban versus rural). Findings demonstrated that an earlier age at menarche is consistently associated with earlier intercourse. Menarche should therefore be used as a target age for reinforcing and expanding adolescent females' knowledge of risks associated with sexual intercourse.

Teaching medicine as a human experience: a patient-doctor relationship course for faculty and first-year medical students.

We developed a required, longitudinal course for first-year medical students that addressed the patient-doctor relationship. Our course linked understanding patients' experiences and perspectives on illness with listening to, talking with, and establishing a rapport with patients while obtaining their medical histories. Learning was enhanced by use of an interdisciplinary faculty and by small-group continuity and faculty mentoring. Our curriculum adapted problem-based, self-directed educational methods to convey medical humanism. We focused on bedside interviewing as the means for exploring patients' social, emotional, and ethical concerns.

Pharmacist participation in the management of incidents involving hazardous materials.

The role of the pharmacist as a hazardous materials consultant is described. Pharmacists in a university-affiliated teaching hospital are contacted by either emergency medical services or the emergency department to assist in the management of incidents involving toxic hazardous materials. These incidents can range from major chemical spills or leaks to long-term exposures involving generalized, nonspecific symptoms. An advanced pharmacy resident in emergency medicine is the primary pharmacy contact for hazardous materials consults. The services provided by the clinical pharmacist include identification of the hazardous materials involved, initiation of special precautions for rescue-squad and hospital-based personnel, clinical assessment of the toxicologic problem, and formulation of therapeutic recommendations. Teaching programs have been developed for pharmacy, nursing, and medical students, hospital employees, and emergency-response agencies. Pharmacy participation in the management of hazardous materials incidents has been well received by emergency department physicians and nurses, as well as by rescue personnel. During the period between January 1 and July 1, 1986, the pharmacy was consulted on 66 hazardous materials incidents. Since pharmacists have traditionally been used as information resources for clinical toxicology questions, it follows that their participation can extend into the field of environmental toxicology, specifically involving hazardous materials. The pharmacist's input as a hazardous materials consultant in our institution has been well received, and we believe that pharmacy departments can play an important role in the management of incidents involving hazardous materials.

Sulindac suppresses tumorigenesis in the Min mouse.

The Min mouse provides a genetically defined model for inherited and sporadic forms of human colorectal tumorigenesis. To test the suitability of this model for the evaluation and optimization of chemopreventive agents, we examined the effects of sulindac on tumorigenesis in Min mice as this compound can inhibit colorectal tumorigenesis in human familial adenomatous polyposis patients. Treatment of Min mice with sulindac in their drinking water (84 mg/l) or diet (167 and 334 p.p.m.) resulted in a significantly decreased average tumor load. The conservation of sulindac activity in the Min mouse provides an opportunity to explore the mechanism of sulindac suppression as well as to test other potential chemopreventive agents.

WAF1, a potential mediator of p53 tumor suppression.

The ability of p53 to activate transcription from specific sequences suggests that genes induced by p53 may mediate its biological role as a tumor suppressor. Using a subtractive hybridization approach, we identified a gene, named WAF1, whose induction was associated with wild-type but not mutant p53 gene expression in a human brain tumor cell line. The WAF1 gene was localized to chromosome 6p21.2, and its sequence, structure, and activation by p53 was conserved in rodents. Introduction of WAF1 cDNA suppressed the growth of human brain, lung, and colon tumor cells in culture. Using a yeast enhancer trap, a p53-binding site was identified 2.4 kb upstream of WAF1 coding sequences. The WAF1 promoter, including this p53-binding site, conferred p53-dependent inducibility upon a heterologous reporter gene. These studies define a gene whose expression is directly induced by p53 and that could be an important mediator of p53-dependent tumor growth suppression.

Simian immunodeficiency virus (SIV)-specific cytotoxic T lymphocytes in gastrointestinal tissues of chronically SIV-infected rhesus monkeys.

Although systemic virus-specific cytotoxic T lymphocyte (CTL) responses are of critical importance in controlling virus replication in individuals infected with human immunodeficiency virus 1 (HIV-1), little is known about this immune response in the gastrointestinal (GI) tract. This study investigated the GI tract CTL response in a nonhuman primate model for HIV-1 infection, simian immunodeficiency virus (SIV)-infected rhesus monkeys. Lymphocytes from duodenal pinch biopsy specimens were obtained from 9 chronically SIVmac-infected rhesus monkeys and GI tract lymphocytes were harvested from the jejunum and ileum of 4 euthanized SIVmac-infected rhesus monkeys. Lymphocytes were also assessed in GI mucosal tissues by in situ staining in histologic specimens. SIVmac Gag-specific CTLs were assessed in the monkeys using the tetramer technology. These GI mucosal tissues of chronically SIVmac-infected rhesus monkeys contained levels of CTLs comparable to those found in the peripheral blood and lymph nodes. The present studies suggest that the CD8(+) CTL response in GI mucosal sites is comparable to that seen systemically in SIVmac-infected rhesus monkeys.