MAP kinase signalling pathways in cancer.

Cancer can be perceived as a disease of communication between and within cells. The aberrations are pleiotropic, but mitogen-activated protein kinase (MAPK) pathways feature prominently. Here, we discuss recent findings and hypotheses on the role of MAPK pathways in cancer. Cancerous mutations in MAPK pathways are frequently mostly affecting Ras and B-Raf in the extracellular signal-regulated kinase pathway. Stress-activated pathways, such as Jun N-terminal kinase and p38, largely seem to counteract malignant transformation. The balance and integration between these signals may widely vary in different tumours, but are important for the outcome and the sensitivity to drug therapy.

Raf kinase inhibitor protein interacts with NF-kappaB-inducing kinase and TAK1 and inhibits NF-kappaB activation.

The Raf kinase inhibitor protein (RKIP) acts as a negative regulator of the mitogen-activated protein (MAP) kinase (MAPK) cascade initiated by Raf-1. RKIP inhibits the phosphorylation of MAP/extracellular signal-regulated kinase 1 (MEK1) by Raf-1 by disrupting the interaction between these two kinases. We show here that RKIP also antagonizes the signal transduction pathways that mediate the activation of the transcription factor nuclear factor kappa B (NF-kappaB) in response to stimulation with tumor necrosis factor alpha (TNF-alpha) or interleukin 1 beta. Modulation of RKIP expression levels affected NF-kappaB signaling independent of the MAPK pathway. Genetic epistasis analysis involving the ectopic expression of kinases acting in the NF-kappaB pathway indicated that RKIP acts upstream of the kinase complex that mediates the phosphorylation and inactivation of the inhibitor of NF-kappaB (IkappaB). In vitro kinase assays showed that RKIP antagonizes the activation of the IkappaB kinase (IKK) activity elicited by TNF-alpha. RKIP physically interacted with four kinases of the NF-kappaB activation pathway, NF-kappaB-inducing kinase, transforming growth factor beta-activated kinase 1, IKKalpha, and IKKbeta. This mode of action bears striking similarities to the interactions of RKIP with Raf-1 and MEK1 in the MAPK pathway. Emerging data from diverse organisms suggest that RKIP and RKIP-related proteins represent a new and evolutionarily highly conserved family of protein kinase regulators. Since the MAPK and NF-kappaB pathways have physiologically distinct roles, the function of RKIP may be, in part, to coordinate the regulation of these pathways.

FRA-1 as a driver of tumour heterogeneity: a nexus between oncogenes and embryonic signalling pathways in cancer.

Tumour heterogeneity is a major factor undermining the success of therapies targeting metastatic cancer. Two major theories are thought to explain the phenomenon of heterogeneity in cancer--clonal evolution and cell plasticity. In this review, we examine a growing body of work implicating the transcription factor FOS-related antigen 1 (FRA-1) as a central node in tumour cell plasticity networks, and discuss mechanisms regulating its activity in cancer cells. We also discuss evidence from the FRA-1 perspective supporting the notion that clonal selection and cell plasticity represent two sides of the same coin. We propose that FRA-1-overexpressing clones featuring high plasticity undergo positive selection during consecutive stages of multistep tumour progression. This model underscores a potential mechanism through which tumour cells retaining elevated levels of plasticity acquire a selective advantage over other clonal populations within a tumour.

PR55α-containing protein phosphatase 2A complexes promote cancer cell migration and invasion through regulation of AP-1 transcriptional activity.

The proto-oncogene c-Jun is a component of activator protein-1 (AP-1) transcription factor complexes that regulates processes essential for embryonic development, tissue homeostasis and malignant transformation. Induction of gene expression by c-Jun involves stimulation of its transactivation ability and upregulation of DNA binding capacity. While it is well established that the former requires JNK-mediated phosphorylation of S63/S73, the mechanism(s) through which binding of c-Jun to its endogenous target genes is regulated remains poorly characterized. Here we show that interaction of c-Jun with chromatin is positively regulated by protein phosphatase 2A (PP2A) complexes targeted to c-Jun by the PR55α regulatory subunit. PR55α-PP2A specifically dephosphorylates T239 of c-Jun, promoting its binding to genes regulating tumour cell migration and invasion. PR55α-PP2A also enhanced transcription of these genes, without affecting phosphorylation of c-Jun on S63. These findings suggest a critical role for interplay between JNK and PP2A pathways determining the functional activity of c-Jun/AP-1 in tumour cells.

Alcohol-related diols cause acute insulin resistance in vivo.

Epidemiological studies suggest that alcohol consumption is an independent risk factor for the development of non-insulin-dependent diabetes mellitus (NIDDM). Alcoholism is known to be associated with increased plasma levels of two novel diols, 2,3-butanediol and 1,2-propanediol, metabolites known to impair insulin action in isolated adipocytes. This study examines whether 2,3-butanediol and 1,2-propanediol have the capacity to impair insulin action acutely in vivo in the rat. Using the euglycemic-hyperinsulinemic clamp, it is shown that the two diols reduce whole-body glucose utilization (by approximately 30%), with the onset of insulin resistance in vivo occurring at plasma concentrations of 2,3-butanediol (33 micromol/L) at least one order of magnitude (P < .001) lower than 1,2-propanediol (432 micromol/L). Tracer methodologies using [U-14C]glucose and 2-deoxy[1-(3)H]glucose indicate that the reduction in whole-body glucose utilization is accompanied by a reduction in glucose uptake and glycogen synthesis in the skeletal muscle and heart. The association between elevated plasma diol levels and insulin resistance demonstrated in this report raises the question of whether there is a link between the high plasma diol levels in alcohol abusers and their increased susceptibility to NIDDM.

Alcoholic myopathy: biochemical mechanisms.

Between one- and two-thirds of all alcohol abusers have impairment of muscle function that may be accompanied by biochemical lesions and/or the presence of a defined myopathy characterised by selective atrophy of Type II fibres. Perturbations in protein metabolism are central to the effects on muscle and account for the reductions in muscle mass and fibre diameter. Ethanol abuse is also associated with abnormalities in carbohydrate (as well as lipid) metabolism in skeletal muscle. Ethanol-mediated insulin resistance is allied with the inhibitory effects of ethanol on insulin-stimulated carbohydrate metabolism. It acutely impairs insulin-stimulated glucose and lipid metabolism, although it is not known whether it has an analogous effect on insulin-stimulated protein synthesis. In alcoholic cirrhosis, insulin resistance occurs with respect to carbohydrate metabolism, although the actions of insulin to suppress protein degradation and stimulate amino acid uptake are unimpaired. In acute alcohol-dosing studies defective rates of protein synthesis occur, particularly in Type II fibre-predominant muscles. The relative amounts of mRNA-encoding contractile proteins do not appear to be adversely affected by chronic alcohol feeding, although subtle changes in muscle protein isoforms may occur. There are also rapid and sustained reductions in total (largely ribosomal) RNA in chronic studies. Loss of RNA appears to be related to increases in the activities of specific muscle RNases in these long-term studies. However, in acute dosing studies (less than 1 day), the reductions in muscle protein synthesis are not due to overt loss of total RNA. These data implicate a role for translational modifications in the initial stages of the myopathy, although changes in transcription and/or protein degradation may also be superimposed. These events have important implications for whole-body metabolism.

Pathogenicity of Salmonella enteritidis phage types 4, 8 and 23 in specific pathogen free chicks.

The pathogenicity of two isolates of Salmonella enterica serovar Enteritidis (SE) phage type (PT) 4, three of PT8 and one of PT23 was investigated in groups of 1-day-old specific pathogen free White Leghorn chicks. Two groups were crop gavaged with each culture but at two different doses. Two additional groups were given Salmonella enterica serovar Pullorum (SP) at similar doses and one further group served as uninoculated controls. Body weights were recorded at 14, 21, and 28 days postinoculation (d.p.i), and mortality was monitored throughout. In most treatment groups, the average body weights were significantly lower than the controls. Birds inoculated with SP had the highest mortality followed by those given SE PT4 of human or chicken origin. At 14 and 21 d.p.i., four chicks from each group were killed and examined for gross lesions. Selected tissues were collected for histopathology and cultured for bacteria. Dead birds had fibrinous exudate in the pericardium and also, in a few, on the liver capsule. They had enlarged livers, sometimes with congestion and white foci. At 7 d.p.i., several birds, especially those inoculated with SE PT4, had retained yolk sacs containing coagulated material. Microscopic lesions of pericarditis, myocarditis, hepatitis, splenitis, peritonitis and enteritis were present at 7 d.p.i. in most birds inoculated with SE, but was greatly variable at 14 d.p.i.. This study shows that 1-day-old SPF chicks are susceptible to various phage types of SE, with yolk-sac infection as the most prominent feature.

Massive acute haemolysis in neonates with glucose-6-phosphate dehydrogenase deficiency.

Three neonates with glucose-6-phosphate dehydrogenase (G6PD) deficiency are described. All three patients suffered an episode of massive acute haemolysis, in the absence of blood group incompatibilities, infection, or ingestion of oxidising agents known to trigger haemolysis. One patient died, but the other two survived after an exchange transfusion. This highlights that G6PD deficiency in the neonatal period may present with severe anaemia in association with hyperbilirubinaemia.

Rehab rounds: overcoming barriers to individualized psychosocial rehabilitation in an acute treatment unit of a state hospital.

Psychiatric rehabilitation begins during the acute stages of a psychiatric disorder and continues throughout the person's lifetime, with the types of services flexibly keyed to the person's phase of illness, needs, and personal goals. During periods of relapse and exacerbation of symptoms, when hospitalization is often required, psychiatric rehabilitation should include the following five objectives: * Clarify how the person's own goals in life, such as a desire for more self-control, freedom of choice, privacy, and time with friends and family, can be served by inpatient treatment and symptom stabilization. * Educate the patient about the nature of his or her illness and how medications work to restore self-control. * Teach the patient about side effects and self-monitoring and negotiating about medication and its effects in a collaborative way with the psychiatrist and other members of the treatment team. * Connect with the family or other natural supports that the person has in the community. * Enable the patient to make appropriate aftercare plans for residential and continuing treatment needs after discharge. When rehabilitation is viewed from the vantage point of these objectives, the inextricable interweaving of "treatment" with "rehabilitation" becomes clear. Treatment and rehabilitation are two sides of the same. It is much easier to integrate psychiatric rehabilitation into more traditional methods of treatment than it is to reorganize a treatment program or facility so that it blends rehabilitation with prevailing treatment imperatives of pharmacotherapy, supervision, and security and safety. In previous Rehab Rounds columns, we have described examples of creative methods for bringing the principles and practices of psychiatric rehabilitation into the treatment milieu (1,2,3). Faced with regulatory criticism from governmental agencies, Dr. Dhillon and his colleagues at Eastern State Hospital in Williamsburg, Virginia, launched a vigorous initiative to bring psychiatric rehabilitation into the forefront of their clinical enterprise. To enable readers to learn from their successful experience and adapt some of the administrative and clinical procedures that worked in Virginia, Dr. Dhillon and Ms. Dollieslager describe the operational details of their odyssey. We believe that their effectiveness in changing a traditional institution can be duplicated in many other places-in units within general hospitals or other community-based settings-as well as in state psychiatric hospitals, where acute treatment has been limited to pharmacotherapy and recreational and diversional activities.

Pathology of avian adenovirus serotypes in the presence of Escherichia coli in infectious-bursal-disease-virus-infected specific-pathogen-free chickens.

Ten strains of adenovirus representing 10 serotypes were administered intratracheally to 3-week-old specific-pathogen-free chickens, which also received 2.5 X 10(5) colony-forming units of a pathogenic Escherichia coli intranasally. Those birds had been inoculated by eye drop at 1 day of age with a virulent infectious bursal disease virus (IBDV). Controls consisted of groups of chickens inoculated with: (a) IBDV and E. coli, (b) IBDV only, (c) E. coli only, and (d) no virus or E. coli. Gross pulmonary alterations at 5 days postinoculation (PI) included congestion and consolidation of one or both lungs of a chick inoculated with adenovirus serotype Ind-C and another inoculated with A-2. Histopathologic alterations in the lungs were those of multifocal interstitial and occasionally diffuse pneumonia. All 10 adenovirus serotypes elicited multifocal or diffuse pneumonia and bronchiolitis in one or more of the five birds per group necropsied at 5 days PI. T-8 and A-2 serotypes induced marked to serve diffuse pneumonia within 5 days; Ind-C, Stein, Tipton, 75-1A, B-3, and X-11 incited a mild diffuse pneumonia. In all groups, the pneumonic lesions were more severe 5 days PI than 12 days PI. Tracheitis was incited by Ind-C, Stein, T-8, and A-2 at 5 days PI; the lesions were minimal to marked in severity. None of the four control groups exhibited gross or histopathologic alterations except the two IBDV-infected groups, which exhibited bursal change.

A comparison of the pathogenicity of four avian reoviruses in chickens.

Four avian reoviruses were orally inoculated into 1-day-old chickens to determine pathogenicity, virus persistence in the intestinal tract, and effects on body weight gains. Avian reoviruses Reo-25 and W3-492 belonged to two separate serotypes, and viruses TC 897 and W3-410 were antigenically related to W3-492. Isolate W3-492, which was highly pathogenic, was very rarely recovered from cloacal swabs collected 2 weeks postinoculation, but inoculated chickens gained significantly less weight (P less than or equal to 0.001) than uninoculated controls during the 5-week test study. Isolate Reo-25 persisted the longest in the intestinal tract, and isolates TC 897 and W3-410, of intermediate persistence, had no significant effect on body weights. There was no apparent correlation between serotype and pathotype of avian reoviruses.

Adenovirus infection associated with respiratory disease in commercial chickens.

The lesions and etiologic agents associated with 13 outbreaks of respiratory disease in commercial chickens were investigated. Adenoviruses were isolated from tracheal and lung tissues of affected chickens in all 13 outbreaks. Escherichia coli was isolated from the lung of an occasional bird. The tracheal specimens were consistently negative for Bordetella avium, but E. coli and occasionally Staphylococcus aureus were isolated. There was also serological evidence in one outbreak, and pathological evidence in another, of a concurrent infectious bursal disease virus (IBDV) infection of chickens affected with the disease. Gross and microscopic alterations in the tracheas and lungs of affected chickens were similar in all outbreaks and consisted of catarrhal tracheitis and occasionally multifocal pneumonia with mononuclear cell infiltrates. Hepatitis and splenitis with heterophil infiltrates occasionally were seen in birds with coliform septicemia. The tracheal and lung lesions in the present investigation were considered primarily of adenovirus etiology, complicated by secondary bacterial infection.

The oncogenic potential of eleven avian adenovirus strains.

Oncogenic potentials of 11 strains of avian adenovirus representing 10 serotypes were investigated in 22 litters of 1-day-old Syrian hamsters (Mesocricetus auratus). Hamsters were inoculated via two routes: subcutaneous dose of 0.12 ml and intracerebral dose of 0.02 ml. Six strains, Indiana-C (Ind-C), chicken embryo lethal orphan (CELO), Stein, 75-1A, B-3, and A-2, induced fibrosarcomas subcutaneously at the inoculation site. Hamsters inoculated intracerebrally with Ind-C, CELO, Stein, B-3, and A-2 developed carcinomas of the choroid plexus. Strain 75-1A virus produced tumors on subcutaneous inoculation but failed to produce tumors on intracerebral inoculation. The Tipton, J-2, T-8, C-2B, and X-11 strains did not produce tumors in hamsters. Uninoculated control hamsters did not develop tumors.

Two outbreaks of colibacillosis in commercial caged layers.

A severe septicemia of Escherichia coli etiology was diagnosed in two houses containing 51,570 and 76,200 layers with mortalities of 6.83% and 4.27%, respectively. Dead birds were removed every other day; however, on occasion, gathering was done on the third day. The disease started in 22-wk-old pullets, and 3 wk later was diagnosed in 82-wk-old birds in an adjoining house. The duration of mortality in house 1 was 12 wk and in house 2 was 13 wk. A non-lactose-fermenting E. coli was isolated. Another outbreak of colibacillosis was diagnosed with 7.8% mortality at a different farm containing 47,000 110-wk-old hens and 10,000 40-wk-old pullets. On this farm the duration of mortality was not provided. Antibiotic treatment failed to reduce losses; chlorine added to the drinking water was effective in controlling the spread of disease and in reducing mortality at both farms.

Pathogenicity of various adenovirus serotypes in the presence of Escherichia coli in chickens.

Ten strains of adenovirus representing 10 serotypes were administered intratracheally to 3-week-old specific-pathogen-free chickens, which also received 2.9 X 10(5) colony-forming units of a pathogenic Escherichia coli intranasally. One group was given only E. coli, and one was retained as an uninoculated control. Gross pathologic alterations post-mortem were minimal and limited to multiple scattered, pale areas in the lungs of an occasional chicken in various groups. Histopathologic changes in the lungs were those of multifocal, interstitial, and occasionally diffuse pneumonia. Moderate to marked interstitial pneumonia was incited by adenovirus strains 75-1A, B-3 A-2, C-2B, and X-11; Ind-C, Stein, Tipton, J-2, and T-8 caused similar but milder lesions. Strains 75-1A, A-2, C-2B, T-8, and X-11 incited moderate to marked multifocal pneumonia; Ind-C, Stein, Tipton, J-2, and B-3 caused mild multifocal pneumonia. In all groups, the pneumonic lesions were more severe 5 days postinoculation than 12 days postinoculation. Bronchiolitis and tracheitis lesions also varied in severity with serotype. A mild hepatitis was seen with serotypes T-8 and 75-1A. Neither the uninoculated control group nor the group inoculated with only E. coli exhibited gross or histopathologic alterations.

Selenium-vitamin E deficiency in captive wild ducks.

Seventy-five ducklings belonging to nine species of wild ducks were kept in a large floor pen. At about 8 weeks of age, a pintail and two shovelers showed clinical signs of anorexia, diarrhea, and leg weakness before death. Gross pathologic alterations included a pale discoloration of gizzard musculature and pale streaks in the leg muscle. Microscopic alterations in gizzard smooth-muscle cells included hyalinization, mineralization of sarcoplasmic debris in necrotic smooth-muscle fibers, and macrophagic invasion and phagocytosis of sarcoplasmic debris. Similar alterations were present in sections of skeletal leg muscle. The pathological lesions were characteristic of selenium-vitamin E deficiency.

Comparative pathogenicity and serogrouping of three Washington isolates of infectious bursal disease virus.

The pathology of three infectious bursal disease virus (IBDV) isolates of Washington poultry origin (WA-678, WA-770, and WA-994) and seven other known IBDV strains (SAL, D-78, MO, OH, Var-A, 2512, and IM) was studied in 3-week-old specific-pathogen-free chickens. Inoculation with IM and 2512 strains resulted in illness and death. No clinical signs or mortality were present with WA-678, WA-770, and WA-994. Macroscopically, bursae were swollen and gelatinous with occasional hemorrhages. Isolate WA-994 caused marked bursal atrophy. Isolate WA-678 elicited moderate bursal pathology. Isolate WA-770 resulted in minimal atrophy. Strains IM and 2512 caused severe bursal atrophy, and strains Var-A and D-78 caused moderate atrophy. Strains SAL, MO, and OH caused no demonstrable bursal atrophy. Results of the cross-neutralization study showed that the three isolates were more closely related to serotype 1 than to serotype 2 IBDV.

Growth of serotypes I and II and variant strains of infectious bursal disease virus in Vero cells.

The growth of five strains of infectious bursal disease virus--three strains of serotype I (SAL, D-78, 2512), one of serotype II (OH), and one variant strain (Variant-A)--were compared in Vero and chicken embryo fibroblast (CEF) cell cultures in order to characterize the replication of different strains of IBDV in Vero cells. For all five virus strains, the latent period in Vero cells ranged from 12 to 18 hr, which was longer than the 4-to-6-hr latent period observed in CEF cultures for strains SAL, D-78, and OH. Virus strains SAL, D-78, and OH, which were examined in both Vero and CEF cultures, also had a more extensive maturation phase and higher yields of virus in Vero than in CEF cultures. Total titers of these viruses of 5.35 to 6.10 log10 TCID50/ml in CEFs occurred 24 to 30 hr postinoculation (PI), although the cytopathic effect (CPE) was not seen until 72 hr PI. By comparison, their total infectious virus titers of 6.85 to 8.35 log10 TCID50/ml in Vero cells occurred from 48 hr PI, coinciding with the appearance of CPE. The growth curve of Variant-A in Vero cells differed from the other viruses by showing steadily rising extracellular and cell-associated virus titers throughout the 72-hr observation period. Only very low titers of Variant-A were obtained in CEF cultures, and thus no growth curve in CEFs was performed.

Viral arthritis in fryers related to reovirus infection in breeders.

In 1983, twenty-two outbreaks of viral arthritis/tenosynovitis were diagnosed in a 6-month period on 18 fryer farms of one commercial operation located in western Washington. The main source of the reovirus infection was traced to a breeder flock that supplied progeny chicks to all of the affected farms.

Detection of avian polyomavirus infection by polymerase chain reaction using formalin-fixed, paraffin-embedded tissues.

Avian polyomavirus infection in psittacines was diagnosed in tissues by the use of polymerase chain reaction (PCR) test. The tissues used in the procedure were either formalin-fixed tissues embedded in paraffin blocks or fresh tissues (heart, liver, and spleen) collected from the psittacines during necropsy. DNA was extracted from these tissues and was tested with the published primers for avian polyomavirus VP1 gene in the PCR that yielded an amplicon of 550 base pair size, which was then visualized by electrophoresis. The amplicon size was consistent with avian polyomavirus. The PCR test was found to be an effective method for identifying avian polyomavirus infection in both formalin-fixed, paraffin-embedded and fresh tissues from psittacine birds of different age groups.