Panlobular emphysema is associated with COPD disease severity: A study of emphysema subtype by computed tomography.

BACKGROUND: Computed tomography has the potential to inform COPD prognosis. We sought to determine associations of emphysema phenotype with clinical parameters including lung function, inflammatory markers, and quality of life. METHODS: Participants of this single-center observational cohort (n = 83) were 40-80 years old, had ≥10 pack-year smoking, and a diagnosis of COPD confirmed by spirometry. All participants had available historic chest CT scans which were systematically reviewed by a single expert radiologist and scored for emphysema subtype, extent, and distribution. Associations between radiographic findings and clinical parameters were determined. RESULTS: Median age of participants was 72 years, median smoking 40 pack-years, and median FEV1 59% predicted. 84% of the participants had radiographic emphysema. Of those, 26% had panlobular emphysema (PLE), 68% centrilobular emphysema (CLE), and 6% paraseptal emphysema (PSE). As compared to the participants with no radiographic emphysema, the presence of PLE-dominant emphysema was associated with a lower BMI (P = 0.012) and greater extent of emphysema (P = 0.014). After adjusting for age, sex, and pack-years smoking history, PLE was associated with greater airflow obstruction by FEV1% (48% vs 71%, P = 0.005), greater symptom burden by CAT score (18 vs 9, P = 0.015), worse quality of life by SGRQ score (43 vs 22, P = 0.025), and more systemic inflammation by erythrocyte sedimentation rate (P = 0.001). CLE- or PSE-dominant emphysema were not similarly associated with clinical features or symptom burden. CONCLUSIONS: The presence of PLE-dominant emphysema was associated with greater extent of emphysema, greater airflow obstruction, increased respiratory symptoms, worse quality of life, and systemic inflammation. Further investigation is indicated to explore the pathogenesis of the PLE phenotype and the prognostic and treatment implications of PLE.

High temporal resolution gastric emptying breath tests in mice.

BACKGROUND: Gastric emptying is a complex physiological process regulating the division of a meal into smaller partitions for the small intestine. Disrupted gastric emptying contributes to digestive disease, yet current measures may not reflect different mechanisms by which the process can be altered. METHODS: We have developed high temporal resolution solid and liquid gastric emptying breath tests in mice using [13 C]-octanoic acid and off axis- integrated cavity output spectroscopy (OA-ICOS). Stretched gamma variate and 2-component stretched gamma variate models fit measured breath excretion data. KEY RESULTS: These assays detect acceleration and delay using pharmacological (7.5 mg/kg atropine) or physiological (nutrients, cold exposure stress, diabetes) manipulations and remain stable over time. High temporal resolution resolved complex excretion curves with 2 components, which was more prevalent in mice with delayed gastric emptying following streptozotocin-induced diabetes. There were differences in the gastric emptying of Balb/c vs C57Bl6 mice, with slower gastric emptying and a greater occurrence of two-phase gastric emptying curves in the latter strain. Gastric emptying of C57Bl6 could be accelerated by halving the meal size, but with no effect on the occurrence of two-phase gastric emptying curves. A greater proportion of two-phase gastric emptying was induced in Balb/c mice with the administration of PYY (8-80 nmol) 60 min following meal ingestion. CONCLUSIONS AND INFERENCES: Collectively, these results demonstrate the utility of high temporal resolution gastric emptying assays. Two-phase gastric emptying is more prevalent than previously reported, likely involves intestinal feedback, but contributes little to the overall rate of gastric emptying.

Inhaled efficacious dose translation from rodent to human: A retrospective analysis of clinical standards for respiratory diseases.

Clinical pharmacologists and toxicologists are often faced with predicting equivalent dosages for humans from biological observations in laboratory animals. Allometric scaling has been used extensively as the basis for extrapolation of drug dosage that might be expected to produce the equivalent biological effects. Allometry is the study of size and its consequences and it is based on the anatomical, physiological, and biochemical similarities between animals. In this review, retrospective analyses have been performed based on data reported in the literature in an attempt to determine the utility of allometric scaling for human dose projections from pre-clinical data for compounds that are delivered by inhalation. The limited pre-clinical efficacy data available on inhaled drugs that are also used clinically supports the current method of scaling using a fixed allometric exponent of 0.67. An example of the utility of the human inhaled dose projections for planning inhaled toxicology studies is also presented.

Evolutionary dynamics of West Nile virus in Georgia, 2001-2011.

From 1999-2001, West Nile virus (WNV) spread throughout the eastern United States (US) and was first detected in Georgia in 2001. To date, the virus has been detected in over 2,500 dead wild bird and mosquito samples from across Georgia. We sequenced the premembrane (preM) and envelope gene (E) (2004 bp) from 111 isolates collected from 2001 to 2011. To assess viral gene flow from other geographic regions in the US, we combined our data with WNV sequences available at the National Center for Biotechnology Information (NCBI) and performed phylogenetic analysis. We found evidence that WNV isolates detected in Chatham County Georgia most likely originated from the Northeastern United States. These results highlight the growing importance of adequate genetic surveillance for monitoring and controlling viruses of public health concern.

Comparative sequence analysis of full-length genome of FIPV at different tissue passage levels.

Feline infectious peritonitis virus (FIPV), an alpha Coronavirus, is the causative agent of a fatal immune mediated disease in cats. It is currently unclear if this virus circulates in the field or develops in felines that are infected with Feline enteric coronavirus. To better understand the genomic changes associated with viral adaptation, we sequenced the complete genomes of FIPV WSU 79-1146 at different tissue passage levels: passage 1, passage 8, and passage 50 tissue culture. Twenty-one amino acid differences were observed in the polyprotein 1a/ab between the different passages. Only one residue change was observed in the spike glycoprotein, which reverted back on subsequent passages, four changes were observed in the 3c protein, and one change was observed in each 3a, small membrane, nucleocapsid and 7a proteins. The mutation rate was calculated to be 5.08-6.3 × 10(-6) nucleotides/site/passage in tissue culture suggesting a relatively stable virus. Our data show that FIPV has a low mutation rate as it is passed in cell culture but has the capacity for change specifically in nsp 2, 3c, and 7b as it is passed in cell culture.

Double-stranded RNA induces molecular and inflammatory signatures that are directly relevant to COPD.

Polyinosinic:polycytidylic acid (poly I:C) is a synthetic analogue of double-stranded (ds)RNA, a molecular pattern associated with viral infections, that is used to exacerbate inflammation in lung injury models. Despite its frequent use, there are no detailed studies of the responses elicited by a single topical administration of poly I:C to the lungs of mice. Our data provides the first demonstration that the molecular responses in the airways induced by poly I:C correlate to those observed in the lungs of chronic obstructive pulmonary disease (COPD) patients. These expression data also revealed three distinct phases of response to poly I:C, consistent with the changing inflammatory cell infiltrate in the airways. Poly I:C induced increased numbers of neutrophils and natural killer cells in the airways, which were blocked by CXCR2 and CCR5 antagonists, respectively. Using gene set variation analysis on representative clinical data sets, gene sets defined by poly I:C-induced differentially expressed genes were enriched in the molecular profiles of COPD but not idiopathic pulmonary fibrosis patients. Collectively, these data represent a new approach for validating the clinical relevance of preclinical animal models and demonstrate that a dual CXCR2/CCR5 antagonist may be an effective treatment for COPD patients.

First report of clinical disease associated with canine distemper virus infection in a wild black bear (Ursus americana).

An approximately 1-yr-old black bear was discovered on the porch of a rural residence in southwestern Pennsylvania on October 26, 2011, where it remained during the day in spite of efforts to frighten it away. The bear exhibited periods of somnolence and sporadic tremors and seizures. It was euthanized by gunshot that evening. Immediately after euthanasia it was observed to have footpads that exuded fluid when compressed. It was submitted for necropsy the next day where roughened footpads were noted. Histologic examination of the brain demonstrated nonsuppurative encephalitis with eosinophilic intranuclear and intracytoplasmic inclusion bodies in neurons. The footpads were thickened and hyperkeratotic. Canine distemper virus (CDV) was detected by immunohistochemistry (IHC) in the brain and footpads, and by reverse transcription polymerase chain reaction (RT-PCR) from the brain tissue. Phylogenetic analysis indicated that the CDV cDNA from the bear had 98.2% nucleotide identity to the Rockborn-Candur vaccine and a canine isolate from 2004 in Missouri, USA, and 97.3% nucleotide identity to a raccoon CDV isolated in 2011 from Tennessee, USA. This represents a first report of CDV as a cause of encephalitis or footpad hyperkeratosis in a wild black bear.

Changes in nonstructural protein 3 are associated with attenuation in avian coronavirus infectious bronchitis virus.

Full-length genome sequencing of pathogenic and attenuated (for chickens) avian coronavirus infectious bronchitis virus (IBV) strains of the same serotype was conducted to identify genetic differences between the pathotypes. Analysis of the consensus full-length genome for three different IBV serotypes (Ark, GA98, and Mass41) showed that passage in embryonated eggs, to attenuate the viruses for chickens, resulted in 34.75-43.66% of all the amino acid changes occurring in nsp 3 within a virus type, whereas changes in the spike glycoprotein, thought to be the most variable protein in IBV, ranged from 5.8 to 13.4% of all changes. The attenuated viruses did not cause any clinical signs of disease and had lower replication rates than the pathogenic viruses of the same serotype in chickens. However, both attenuated and pathogenic viruses of the same serotype replicated similarly in embryonated eggs, suggesting that mutations in nsp 3, which is involved in replication of the virus, might play an important role in the reduced replication observed in chickens leading to the attenuated phenotype.

Older people's cardiac responses as indicators of stress in familiar and unfamiliar environments.

Little is known about older people's physiological and emotional responses to environmental triggers. We examined this by estimating cardiac stress from heart rate variability (HRV). Thirty-eight participants were monitored while observing environmental scenarios at familiar and unfamiliar locations. Image scenarios included pedestrian and driving scenes in a random order. HRV indices including heart rate (HR), QT variability index (QTVI) and Total HRV Power (TP) were quantified. Familiar locations were associated with higher HR (p<.0005) and lower TP (p=.005) than unfamiliar locations, suggesting they were more stressful. HRV responses to pedestrian and driving scenarios indicated that stress was not influenced by either the type of image scenario or the order of image presentation. There were no gender-related differences in cardiac responses. HRV is a useful surrogate of cardiac stress when assessing older people's responses to their environments.

CXCR2 antagonists for the treatment of pulmonary disease.

Chemokines have long been implicated in the initiation and amplification of inflammatory responses by virtue of their role in leukocyte chemotaxis. The expression of one of the receptors for these chemokines, CXCR2, on a variety of cell types and tissues suggests that these receptors may have a broad functional role under both constitutive conditions and in the pathophysiology of a number of acute and chronic diseases. With the development of several pharmacological, immunological and genetic tools to study CXCR2 function, an important role for this CXC chemokine receptor subtype has been identified in chronic obstructive pulmonary disease (COPD), asthma and fibrotic pulmonary disorders. Interference with CXCR2 receptor function has demonstrated different effects in the lungs including inhibition of pulmonary damage induced by neutrophils (PMNs), antigen or irritant-induced goblet cell hyperplasia and angiogenesis/collagen deposition caused by lung injury. Many of these features are common to inflammatory and fibrotic disorders of the lung. Clinical trials evaluating small molecule CXCR2 antagonists in COPD, asthma and cystic fibrosis are currently underway. These studies hold considerable promise for identifying novel and efficacious treatments of pulmonary disorders.

The incidence of deep prosthetic infections in a specialist orthopaedic hospital: a 15-year prospective survey.

The Control of Infection Committee at a specialist orthopaedic hospital prospectively collected data on all episodes of bacteriologically-proven deep infection arising after primary hip and knee replacements over a 15-year period from 1987 to 2001. There were 10 735 patients who underwent primary hip or knee replacement. In 34 of 5947 hip replacements (0.57%) and 41 of 4788 knee replacements (0.86%) a deep infection developed. The most common infecting micro-organism was coagulase-negative staphylococcus, followed by Staphylococcus aureus, enterococci and streptococci. Of the infecting organisms, 72% were sensitive to routine prophylactic antimicrobial agents. Of the infections, 29% (22) arose in the first three months following surgery, 35% between three months and one year (26), and 36% (27) after one year. Most cases were detected early and treated aggressively, with eradication of the infection in 96% (72). There was no significant change in the infection rate or type of infecting micro-organism over the course of this study. These results set a benchmark, and importantly emphasise that only 64% of peri-prosthetic infections arise within one year of surgery. These results also illustrate the advantages of conducting joint replacement surgery in the isolation of a specialist hospital.

Mutational analysis of the N-terminus in Schistocerca gregaria ion-transport peptide expressed in Drosophila Kc1 cells.

The functions of the 6-7 amino acid N-terminal domain conserved in insect and crustacean members of the hyperglycemic hormone (CHH) family were assayed by site-directed mutagenesis of Schistocerca gregaria ion-transport peptide (SchgrITP). Mutant peptides were expressed in Drosophila Kc1 cells and tested in a biological assay measuring stimulation of active Cl(-) transport across the locust ileum. We exchanged the N-terminal domain of SchgrITP with that of the shrimp Penaeus japonicus hyperglycemic hormone leaving the remainder of SchgrITP intact. The chimeric peptide was completely inactive in the ileal bioassay, showing that the N-terminus of SchgrITP is essential and that the 2 amino acids (phenylalanine-3 and aspartate-4) conserved in the shrimp and locust peptides are not sufficient for function. We made all possible alanine substitutions in the SchgrITP N-terminal domain. Only phenylalanines 2 and 3 were essential for function in the locust ileal bioassay. All N-terminal mutations were cleaved correctly from the prepropeptide, and expressed in similar concentrations as wild-type ITP suggesting the specific amino acids are not essential for these functions. Post-translational modification may explain a minor ITP isomorph observed in Drosophila Kc1 cell expression. Alanine substitution at position 2 produced a weak ITP antagonist. These structure-function studies, the first for any member of the CHH family, show that both conserved and unconserved amino acids contribute to SchgrITP ion-transport function and that the conserved aspartate in position 4 is required for a yet uncharacterized function.

Fibrin stimulates platelets to increase factor VIIIa binding site expression.

Factor (F)VIII functions as an enzymatic cofactor on the membranes of stimulated platelets. However, thrombin stimulates platelets to express only a small number of binding sites for FVIII. We wished to determine whether molecules that are likely to be present in a developing thrombus stimulate platelets to up-regulate FVIII binding site expression. Flow cytometry was utilized to measure binding of fluorescein-labeled FVIIIa to activated platelets and a FXase assay was utilized to measure platelet-dependent function. Various agonists as well as normal and mutant fibrinogens and fibrin were evaluated as co-stimuli. Thrombin-stimulated platelets expressed 214 +/- 67 binding sites for thrombin-activated FVIII (FVIIIa) and none of the established soluble agonists enhanced binding site exposure. However, the presence of 5 micro g mL(-1) fibrin increased the number of FVIIIa binding sites/platelet three- to eight-fold (1470 +/- 130, range 600-1800) with a parallel increase in platelet-based FXase assay. Binding site up-regulation was not stimulated by fibrinogen and was blocked by inhibitors of GPIIbIIIa. Mutant fibrin lacking the gamma-chain C-terminal four residues was ineffective while fibrin with altered RGD sequences did stimulate expression of FVIIIa binding sites indicating that co-stimulation is mediated by the fibrin gamma-chain termini. Fibrin-enhanced expression of FVIIIa binding sites was not supported by D364H fibrin, which does not aggregate normally, and was blocked by the GPRP peptide, which inhibits fibrin polymerization. Polymerized fibrin can function as a platelet co-stimulus, up-regulating expression of binding sites for FVIIIa.

Weight gain during adjuvant and neoadjuvant chemotherapy for breast cancer: an audit of 100 women receiving FEC or CMF chemotherapy.

Weight gain is reported as a common finding in patients treated for breast cancer but its aetiology appears to be complex. The aim of this study was to investigate the incidence and degree of weight gain during chemotherapy and to examine possible contributory factors. Data were collected on 100 women treated with CMF or FEC chemotherapy. The mean change in weight was +3.68 kg (P<0.001). 64% of patients gained more than 2 kg in weight, 31% maintained a stable weight (within + or - 2 kg) and 5 patients lost more than 2 kg. Approximately 1/3 of patients (33) gained more than 5 kg and 6 patients gained more than 10 kg in weight. The majority of patients (85%) received steroids as antiemetics but no effect of steroid dose was seen on the level of weight change. No significant differences in weight gain were seen in patients receiving tamoxifen (37%) compared with those not taking it. Similarly, menopausal status did not appear to be a significant factor influencing weight gain. In summary, a high incidence of weight gain was found. The literature on weight gain in breast cancer and possible interventions to avoid weight gain are discussed.

A new technique for posterior cruciate ligament reconstruction.

We describe a new technique for arthroscopically assisted posterior cruciate ligament reconstruction. We use the TransFix (Arthrex, Naples, FL) pin to provide a single and strong tibial attachment point for the 4-stranded semitendinosus and gracilis tendons. The TransFix technique for anterior cruciate ligament reconstruction has been found to give better initial fixation strength than other techniques using semitendinosus and gracilis grafts, and it provides adequate graft length for secure fixation. Strong and secure graft fixation in knee ligament reconstruction is very important for early and fast rehabilitation.

7-Ketocholesterol forms crystalline domains in model membranes and murine aortic smooth muscle cells.

7-Ketocholesterol (7-keto) is one of the major oxygenated products found in oxidized low-density lipoproteins (LDL) and in atherosclerotic plaque, where it is believed to play a role in arterial pathology. We hypothesize that direct membrane effects independent of receptor binding may mediate its biological activity. To test this, small-angle x-ray diffraction approaches were used to examine the interactions of 7-keto with other membrane components in well-defined lipid vesicles and in murine aortic smooth muscle cell membranes. These data were compared with the interactions of 25-hydroxycholesterol (25-OHC) and cholesterol. Replacement of cholesterol with 7-keto in lipid vesicles produced distinct changes in membrane structure, including a marked increase in molecular volume associated with the hydrocarbon core (+/-0-8 A from the bilayer center). Additionally, there was an increase in electron density associated with the upper acyl chain region (+/-9-21 A), corresponding to the bilayer location of the steroid nucleus of 7-keto. In contrast, 25-OHC did not appear to intercalate into the membrane hydrocarbon core and did not form separate domains. Cells grown in the presence of the 7-keto developed extracellular crystals concomitant with the formation of membrane domains having a unit cell periodicity of 35.4 or 1.4 A greater than measured with cholesterol. Domains were formed within 4 h and persisted up to 72 h, after which cells showed signs of declining viability. We conclude that 7-keto is found in a membrane location distinct from cholesterol, does not condense phospholipids as efficiently as cholesterol and is able to self-associate into discrete intrabilayer domains. While these domains may decrease its cytotoxicity by inducing the formation of sterol crystals in smooth muscle cells, they may, in a broader capacity, contribute to the sterol crystals found in advanced atherosclerotic lesions.

Rising incidence and prevalence of herpes simplex type 2 infection in a cohort of 26 year old New Zealanders.

OBJECTIVES: To examine changes in the incidence and prevalence of herpes simplex type 2 (HSV-2) infection in a birth cohort of 26 year old New Zealanders in whom seroprevalence had been measured at 3.4% at age 21. METHODS: Sera from 869 cohort members were tested using an indirect IgG enzyme linked immunoassay specific to the HSV-2 glycoprotein G. Serological results were compared with detailed sexual histories. RESULTS: In all, 96 participants (11%) were seropositive for HSV-2, including at least 56 who seroconverted after their 21st birthday. Among those known to be seronegative at age 21, the annual seroconversion rate was 13.5 cases per 1000 per year, compared with 8.1 cases per 1000 per sexually active year before age 21. New infections were associated with female sex and an early age of first intercourse. The average rate of partner change was lower in the cohort after age 21, and was only modestly increased among those who acquired new HSV-2 infections between ages 21 and 26. CONCLUSIONS: HSV-2 seroprevalence has risen sharply in this sexually active cohort, despite a reduction in the overall level of partner change. Increased rates of HSV-2 acquisition after age 21 may be due to a higher prevalence of infection in the pool of potential partners encountered during the third decade of life. Factors related to partner choice may have more influence on the risk of HSV-2 infection than the number of sexual partners alone.

Regulation of vocal fold transepithelial water fluxes.

Vocal fold hydration is critical to phonation. We hypothesized that the vocal fold generates bidirectional water fluxes, which are regulated by activity of the Na(+)-K(+)- ATPase. Western blots and immunohistochemistry demonstrated the presence of the alpha-subunit Na(+)-K(+)-ATPase in the canine vocal fold (n = 11). Luminal cells, basal and adjacent one to two layers of suprabasal cells within stratified squamous epithelium, were immunopositive, as well as basolateral membranes of submucosal seromucous glands underlying transitional epithelia. Canine (n = 6) and ovine (n = 14) vocal fold mucosae exhibited transepithelial potential differences of 8.1 +/- 2.8 and 9.3 +/- 1.3 mV (lumen negative), respectively. The potential difference and short-circuit current (ovine = 31 +/- 4 microA/cm(2); canine = 41 +/- 10 microA/cm(2)) were substantially reduced by luminal administration of 75 microM acetylstrophanthidin (P < 0.05). Ovine (n = 7) transepithelial water fluxes decreased from 5.1 +/- 0.3 to 4.3 +/- 0.3 microl x min(-1) x cm(-2) from the basal to luminal chamber and from 5.2 +/- 0.2 to 3.9 +/- 0.3 microl x min(-1) x cm(-2) from the luminal to basal chamber by luminal acetylstrophanthidin (P < 0.05). The presence of the Na(+)-K(+)-ATPase in the vocal fold epithelium and the electrolyte transport derived from its activity provide the intrinsic mechanisms to regulate cell volume as well as vocal fold hydration.

Bidirectional transepithelial water transport: chloride-dependent mechanisms.

We hypothesized that inhibition and activation of basolateral to luminal chloride transport mechanisms were associated with respective decreases and increases in basolateral to luminal water fluxes. The luminal to basolateral (J(W)(L-->B)) and basolateral to luminal (J(W)(B-->L)) water fluxes across ovine tracheal epithelia were measured simultaneously. The mean J(W)(L-->B) (6.5 microl/min/cm(2)) was larger than J(W)(B-->L) (6.1 microl/min/cm(2)). Furosemide reduced J(W)(B-->L) from 6.0 to 5.6 microl/min/cm(2). Diphenylamine-2-carboxylate (DPC) reduced J(W)(B-->L) from 7.9 to 7. 3 microl/min/cm(2) and reduced the membrane potential difference by 38%. Furosemide together with DPC decreased J(W)(L-->B) by 30% and J(W)(B-->L) by 15%. Norepinephrine increased J(W)(B-->L) from 4.9 to 6.0 microl/min/cm(2). Neuropeptide Y in the presence of norepinephrine decreased J(W)(L-->B) (6.4 to 5.2 microl/min/cm(2)) and returned J(W)(B-->L) to its baseline value. Vasopressin increased J(W)(B-->L) from 4.1 to 5.1 microl/min/cm(2). Endothelin-1 induced a simultaneous increase in J(W)(B-->L) (7.0 to 7.7 microl/min/cm(2)) and decrease in J(W)(L-->B) (7.4 to 6.4 microl/min/cm(2)); and decreased the membrane resistance. These data indicate that in tracheal epithelia under homeostatic conditions J(W)(B-->L) has a approximately 15% actively coupled component. Consistent with our hypothesis, inhibition and receptor-induced stimulation of chloride effluxes were associated with decreases and increases in J(W)(B-->L), respectively. However, as inhibition of transcellular chloride transport always decreased J(W)(L-->B) more than J(W)(B-->L), reducing transepithelial chloride transport did not result in less water being transported into the airway lumen.

Mutational analysis of the C-terminus in ion transport peptide (ITP) expressed in Drosophila Kc1 cells.

Ion transport peptide (ITP) stimulates Cl(-) transport (measured as short-circuit current, I(sc)) and fluid reabsorption in Schistocerca gregaria ilea. We report that Drosophila Kc1 cells transfected with preproITP cDNA secrete a peptide (KcITP(75)) that, while cleaved correctly at the N-terminus, had reduced (10-fold) stimulatory activity on ileal I(sc) compared to both native ITP (ScgITP) and synthetic ITP (synITP). We provide evidence that the reduced activity of KcITP(75) is due to incomplete processing of the C-terminal sequence LGKK (KcITP(75)) to L-amide. In support of this, in vitro amidation of glycine extended ITP (i.e., KcITP(73) ending in LG) but not KcITP(75) (ending in LGKK) significantly increased specific activity in the bioassay. Further evidence for C-terminus involvement includes complete loss of stimulation by truncated mutants (e.g., KcITP(71) which lacks LGKK) and a mutant in which alanine is substituted for the terminal glycine in KcITP(73). Moreover a natural homologue (KcITP-L, which differs only in the C-terminal sequence) expressed by Kc1 cells does not stimulate ileal I(sc). Rather KcITP-L acts as a weak ITP antagonist, as does the truncated mutant KcITP(71). KcITP(70) has no antagonistic effect. A short synthetic peptide fragment of the C-terminus (VEIL-amide) does not stimulate ileal I(sc), indicating that other regions of ITP are also essential to biological activity. Arch.