p19(Arf) is required for the cellular response to chronic DNA damage.

The p53 tumor suppressor is a stress sensor, driving cell cycle arrest or apoptosis in response to DNA damage or oncogenic signals. p53 activation by oncogenic signals relies on the p19(Arf) tumor suppressor, while p53 activation downstream of acute DNA damage is reported to be p19(Arf)-independent. Accordingly, p19(Arf)-deficient mouse embryo fibroblasts (MEFs) arrest in response to acute DNA damage. However, p19(Arf) is required for replicative senescence, a condition associated with an activated DNA damage response, as p19(Arf)-/- MEFs do not senesce after serial passage. A possible explanation for these seemingly disparate roles for p19(Arf) is that acute and chronic DNA damage responses are mechanistically distinct. Replicative senescence may result from chronic, low-dose DNA damage responses in which p19(Arf) has a specific role. We therefore examined the role of p19(Arf) in cellular responses to chronic, low-dose DNA-damaging agent treatment by maintaining MEFs in low oxygen and administering 0.5 G y γ-irradiation daily or 150 µM hydroxyurea, a replication stress inducer. In contrast to their response to acute DNA damage, p19(Arf)-/- MEFs exposed to chronic DNA damage do not senesce, revealing a selective role for p19(Arf) in senescence upon low-level, chronic DNA damage. We show further that p53 pathway activation in p19(Arf)-/- MEFs exposed to chronic DNA damage is attenuated relative to wild-type MEFs, suggesting a role for p19(Arf) in fine-tuning p53 activity. However, combined Nutlin3a and chronic DNA-damaging agent treatment is insufficient to promote senescence in p19(Arf)-/- MEFs, suggesting that the role of p19(Arf) in the chronic DNA damage response may be partially p53-independent. These data suggest the importance of p19(Arf) for the cellular response to the low-level DNA damage incurred in culture or upon oncogene expression, providing new insight into how p19(Arf) serves as a tumor suppressor. Moreover, our study helps reconcile reports suggesting crucial roles for both p19(Arf) and DNA damage-signaling pathways in tumor suppression.

The pro-longevity gene FoxO3 is a direct target of the p53 tumor suppressor.

FoxO transcription factors have a conserved role in longevity, and act as tissue-specific tumor suppressors in mammals. Several nodes of interaction have been identified between FoxO transcription factors and p53, a major tumor suppressor in humans and mice. However, the extent and importance of the functional interaction between FoxO and p53 have not been fully explored. Here, we show that p53 regulates the expression of FoxO3, one of the four mammalian FoxO genes, in response to DNA damaging agents in both mouse embryonic fibroblasts and thymocytes. We find that p53 transactivates FoxO3 in cells by binding to a site in the second intron of the FoxO3 gene, a genomic region recently found to be associated with extreme longevity in humans. While FoxO3 is not necessary for p53-dependent cell cycle arrest, FoxO3 appears to modulate p53-dependent apoptosis. We also find that FoxO3 loss does not interact with p53 loss for tumor development in vivo, although the tumor spectrum of p53-deficient mice appears to be affected by FoxO3 loss. Our findings indicate that FoxO3 is a p53 target gene, and suggest that FoxO3 and p53 are part of a regulatory transcriptional network that may have an important role during aging and cancer.

Preliminary evidence for the metabolism of benzo(a) pyrene by Plantago lanceolata.

Polycyclic aromatic hydrocarbons (PAHs) have long been recognised as potential carcinogens in animals in which biotransformation into reactive metabolites can lead to DNA damage. In animals PAHs metabolism mainly occurs in hepatic microsomes and is associated with the cytochrome p-450 mediated mixed functional oxidase (MFO) system. PAH metabolism in plants has been shown to occur via a similar enzyme system, but has received relatively little attention. This study is looking at how the plant species Plantago lanceolata metabolizes benzo(a)pyrene (B(a)P), which is one of the PAHs whose metabolism has been studied extensively in animals. The aim of the work is to establish firstly that the B(a)P is taken up and secondly that it is biotransformed by the plant to products possibly similar to those found in animals. This work is achieved by using C-14-B(a)P along with whole body autoradiography, scintillation analysis and chromatography techniques to locate the B(a)P and its metabolites.

Systemic inflammatory response syndrome, sepsis, and multiple organ dysfunction.

Companion animals with sepsis and multiple organ dysfunction can be the most challenging of all patients to treat. Current research in humans and laboratory models offers some exciting insights into the pathophysiology behind some of our most frustrating clinical challenges. This article applies several current concepts to a clinical case of pancreatitis and secondary sepsis to illustrate some of the cardiovascular, immune, and coagulation abnormalities commonly seen.

Generation of a selective 5-HT3B subunit-recognising polyclonal antibody; identification of immunoreactive cells in rat hippocampus.

The present study generated a polyclonal antibody (AP86/3) that recognises a peptide sequence of the h5-HT(3B) receptor subunit. Western blot analysis of homogenates prepared from cell lines expressing either homomeric (h5-HT(3A)) or heteromeric (h5-HT(3A/3B)) receptors, as well as immunocytochemical studies with the same cell lines, indicated that AP86/3 recognised, selectively, the 5-HT(3B) subunit. Immunohistochemical labelling was also apparent in cells in the rat hippocampus that displayed the distribution and morphology of interneurones.

Pharmacological comparison of human homomeric 5-HT3A receptors versus heteromeric 5-HT3A/3B receptors.

The present study determined the detailed pharmacological profile of heterologously expressed human (h) homomeric 5-HT3A receptors in direct comparison to heteromeric h5-HT3A/3B receptors. The very minor differences in their respective pharmacological profiles indicates that the 5-HT3B receptor subunit alters, predominantly, the biophysical rather than the pharmacological properties of the 5-HT3 receptor.

Severe sepsis in cats: 29 cases (1986-1998).

OBJECTIVE: To document the clinical, clinicopathologic, and pathologic findings in cats with severe sepsis, identify abnormalities unique to this species, and identify criteria that could be used antemortem to diagnose the systemic inflammatory response syndrome in cats. DESIGN: Retrospective study. ANIMALS: 29 cats confirmed to have severe sepsis at necropsy. PROCEDURE: Pertinent history, physical examination findings, and results of hematologic and biochemical testing were extracted from medical records. RESULTS: Clinical diagnoses included pyothorax, septic peritonitis, bacteremia secondary to gastrointestinal tract disease, pneumonia, endocarditis, pyelonephritis, osteomyelitis, pyometra, and bite wounds. Physical examination findings included lethargy, pale mucous membranes, poor pulse quality, tachypnea, hypo- or hyperthermia, signs of diffuse pain on abdominal palpation, bradycardia, and icterus. Clinicopathologic abnormalities included anemia, thrombocytopenia, band neutrophilia, hypoalbuminemia, low serum alkaline phosphatase activity, and hyperbilirubinemia. Necropsy findings included multi-organ necrosis or inflammation with intralesional bacteria. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that severe sepsis in cats is characterized by lethargy, pale mucous membranes, signs of diffuse abdominal pain, tachypnea, bradycardia, weak pulses, anemia, hypoalbuminemia, hypothermia, and icterus. Recognition of this combination of clinical findings should facilitate the diagnosis of severe sepsis in cats.

Postoperative management of the emergency surgery small animal patient.

Hypovolemia, hypothermia, and hypotension are common postoperative findings that predispose the critically ill patient to secondary complications. This patient population is especially vulnerable to sepsis, hypoxia, and immune dysfunction. Careful monitoring is essential for early recognition of potentially life-threatening physiologic derangements. Early and aggressive intervention may help minimize systemic insult before it progresses to acute respiratory distress syndrome, acute renal failure, disseminated intravascular coagulation, or multiple organ failure.

Severe neurologic sequelae in a dog after treatment of hypoadrenal crisis.

A 3-year-old mixed-breed dog was evaluated for lethargy, weakness, anorexia, and vomiting. The dog was dehydrated, hyponatremic, hypochloremic, and hypoglycemic. Results of an ACTH stimulation test indicated hypoadrenocorticism. Treatment to restore cardiovascular stability and serum electrolyte balance caused serum sodium concentration to increase by 32 mEq/L within 48 hours, and the dog developed severe neurologic signs that persisted for approximately 3 weeks. Magnetic resonance imaging revealed cerebrocortical lesions on day 6 and more severe lesions, including diffuse atrophy of the cerebral hemispheres, at 23 weeks after initial evaluation; however, the dog recovered complete neurologic function. Serum sodium concentration should be monitored during treatment for hypoadrenal crisis to avoid rapid increases that can cause CNS damage.

Use of peer role models to increase Pap smear and mammogram screening in Mexican-American and black women.

We evaluated the use of a community-based intervention model to increase Pap smear and mammogram screening in minority women. The community programs were based on the A Su Salud (To Your Health) model, which includes the presentation of positive role models in the media and positive social reinforcement by community volunteers. Mexican-American women 40-70 years of age in Corpus Christi, Texas, and black women 40-70 years of age in Galveston, Texas, were targeted. Role models from the target community told their personal stories about cancer screening through local media (television, radio, and newspapers). Volunteers reinforced media messages among the target audience by distributing newsletters. Preintervention and postintervention cross-sectional surveys among the target group measured relative increases in knowledge, attitude, and practice. Pap smear use (past two years) was not substantially improved. Mammogram use (past two years) increased 57% among Mexican-American women (prevalence rate ratio [PRR] = 1.57; 90% confidence interval [CI] = 1.19, 2.08) and 30% among black women (PRR = 1.30; 90% CI = 1.04, 1.63). We demonstrated the feasibility of implementing a peer role model intervention at the local health department level. The improvement in mammogram use among high-risk women is encouraging, but further controlled research is needed to test the effectiveness and cost-efficiency of this approach.

Local health departments implement a theory-based model to increase breast and cervical cancer screening.

Su Vida, Su Salud/Your Life, Your Health is a community program to increase participation in breast and cervical cancer screening. This program illustrates the diffusion of an innovative outreach strategy from a research environment to two local health departments. The program uses A Su Salud, the communication model in which positive role models are featured in the media, and community volunteers who give positive social reinforcement. Local health departments in Galveston and Corpus Christi, TX, were selected to implement the model because of excessive mortality rates in the region and the departments' adequate level of resources and commitment. Over an 18-month period, 54 stories of role models appeared in the media in Corpus Christi, and 60 appeared in Galveston. There were 490 volunteers active in Corpus Christi and 279 in Galveston. Of 365 inquiries to the Corpus Christi program, 35 percent were from the Mexican American target group of women ages 40-70. Of 1,457 women who contacted the Galveston program, 9 percent were from the African American target group ages 40-70. Local health departments are well-suited to participation in cancer control research applications because they have direct access to high-risk populations and have the authority to institutionalize effective interventions.

A note on the isolation and propagation of lytic phages from Streptococcus uberis and their potential for strain typing.

Thirty-eight of 98 strains of Streptococcus uberis were shown to be carrying lysogenic phage. Although propagating strains were rare, host modification by field strains sensitive to phage was used to increase the lytic spectra. When 120 nationally-collected strains were challenged with 25 phages, selected on the basis of differing lytic spectra and propagating strains, 30% were susceptible to at least one phage, increasing to 42% when 480 strains from a single farm were considered. A typing system based on susceptibility to lytic phage was considered feasible.

Identification during the postpartum period of infants who are at high risk of child maltreatment.

To determine whether clinicians correctly identify newborn infants who are at high risk of child maltreatment, we examined the outcomes of high-risk and non-high-risk children. Infants who were born at Yale-New Haven Hospital from 1979 to 1981 and who were referred by clinicians during the postpartum period to the hospital's child abuse registry because they were considered at high risk of child abuse or neglect became the high-risk group. For each high-risk infant, a comparison infant was selected and matched according to date of birth, gender, race, and method of payment for the hospitalization. For both groups, the occurrence of maltreatment was ascertained by reviewing the medical records until the child's fourth birthday. Each injury for which medical care was sought was classified into one of seven categories (from definite child abuse to accident) by a pediatrician who was unaware of the child's risk status. Information also was recorded about nonorganic failure to thrive and changes in the child's caretaker. Maltreatment (defined as abuse or neglect) occurred more frequently in the high-risk group than the comparison group (adjusted matched odds ratio = 4.3; 95% confidence interval = 1.41, 6.93; p less than 0.001), as did poor weight gain from a nonorganic cause (matched odds ratio = 7.0; 95% confidence interval = 1.59, 30.79; p less than 0.01) and changes in the child's caretaker (matched odds ratio = 9.0; 95% confidence interval = 3.80; 20.55; p less than 0.001). We conclude that as early as the postpartum period, clinicians can identify some families who are at high risk of maltreatment and other major adverse outcomes resulting from poor parenting.

Development of lymphocytotoxic and platelet reactive antibodies: a prospective study in patients with acute leukaemia.

Lymphocytotoxic (LCT) and platelet reactive (PR) antibody (Ab) responses were serially determined in 49 patients with acute leukaemia. LCTAb were found in 20 patients and occurred in 13 patients with acute myeloid leukaemia and 7 patients with acute lymphoblastic leukaemia. Four differing patterns of LCTAb responses could be defined. Thirteen of 22 subjects showed marked reduction or loss of LCTAb. Indirect platelet immunofluorescence, measured by flow cytometry, provided the most convenient means of detecting PRAb which were found in 11 subjects and generally showed moderate or weak reactivity.

The protective efficacy of Haemophilus b polysaccharide vaccine.

To assess the protective efficacy of the Haemophilus b polysaccharide vaccine, a case-control study was conducted in Connecticut, Dallas County, and greater Pittsburgh. Seventy-six children 24 to 72 months of age who had H influenzae type b isolated from normally sterile sites from August 1985 through July 1987 were enrolled. For each case two controls, matched by age and place of residence, were randomly selected from the records of all live births in the area. Antecedent receipt of the Haemophilus b polysaccharide vaccine was ascertained from the records of all physicians and clinics at which the subjects received medical care. Overall, 12% of the cases and 39% of the controls had received the vaccine. The estimate of the protective efficacy of the vaccine was 88% overall (95% confidence interval, 74% to 96%) and 91% (95% confidence interval, 71% to 99%), 92% (95% confidence interval, 76% to 99%), and 81% (95% confidence interval, 45% to 96%) in Connecticut, Dallas County, and greater Pittsburgh, respectively. The estimates were not substantially affected by adjusting with logistic regression for differences between the cases and controls in race and the attendance of group day care. We conclude that the Haemophilus b polysaccharide vaccine is highly effective in these areas among children who receive the vaccine when they are 24 months of age or older.

Job satisfaction and perception of social climate in a mental health facility.

21 members of a staff process group at a mental health treatment and research center were administered the Job Descriptive Index and the Work Environmental Scale once a month for a 3-mo. period. A two-day analysis of variance (job satisfaction x time) was computed for each of the 10 subscales. The three administrations did not significantly vary on any of the dimensions, nor were significant interaction effects found. Subjects high in satisfaction significantly differed from those who scored low on the dimensions of involvement and peer cohesion, staff support, autonomy, and innovation. However, no significant differences were found on the dimensions of task orientation, work pressure, clarity, control, or physical comfort. Results were discussed in terms of the importance of the variables, relationship and responsibility, to job satisfaction in a mental health setting.