Biomedical subjects
S J Johnson
Publications and source records attributed to S J Johnson.
Identification and analysis of homoeologous segments of the genomes of rice and Arabidopsis thaliana.
Using contiguous genomic DNA sequences of Arabidopsis thaliana, we were able to identify a region of conserved structure in the genome of rice. The conserved, and presumptive homoeologous segments, are 194 kb and 219-300 kb in size in Arabidopsis and rice, respectively. They contain five homologous genes, distinguished in order by a single inversion. These represent the first homoeologous segments identified in the genomes of a dicot and a monocot, demonstrating that fine-scale conservation of genome structure exists and is detectable across this major divide in the angiosperms. The conserved framework of genes identified is interspersed with non-conserved genes, indicating that mechanisms beyond segmental inversions and translocations need to be invoked to fully explain plant genome evolution, and that the benefits of comparative genomics over such large taxonomic distances may be limited.
Macrophage and hepatic stellate cell responses during experimental hepatocarcinogenesis.
The aim of the study was to assess the monocyte/macrophage and hepatic stellate cell responses during experimental diethylnitrosamine (DEN)-induced hepatocarcinogenesis. Diethylnitrosamine (50mg/L) was administered to 39 rats for 10 weeks; liver tissue was obtained at weeks 10, 16 and 19. In this model, necroinflammatory damage occurs during the period of DEN administration but thereafter subsides; dysplastic nodules and carcinomas subsequently develop. Monocytes/ macrophages were detected immunohistochemically using ED1 and ED2 monoclonal antibodies; hepatic stellate cells (HSC) were detected using antibodies to alpha-smooth muscle actin (alpha-SMA) (activated HSC) and glial fibrillary acidic protein (GFAP). Parenchymal ED1- and ED2-positive monocytes/macrophages and alpha-SMA-positive HSC increased at week 10 when there was ongoing DEN-induced necroinflammatory activity. ED1- and ED2-positive cells were also prominent at weeks 16 and 19, particularly around the periphery of dysplastic and carcinomatous nodules, with occasional macrophages between dysplastic hepatocytes. alpha-SMA-positive HSC were present within sinusoids between dysplastic cells and were more abundant at weeks 16 and 19 than in control or week 10 animals. Activated HSC were prominent in fibrous septa around and within dysplastic and carcinomatous nodules at weeks 16 and 19. In contrast, GFAP-positive HSC did not accumulate in developing septa or within dysplastic and carcinomatous nodules. We have demonstrated changes in the monocyte/ macrophage and HSC populations during the development of hepatocellular dysplasia and carcinoma at time points when there is little necroinflammatory activity; this may therefore represent a host response to hepatocyte dysplasia. The HSC activation may be mediated, in part, by monocyte/ macrophage-derived factors, but we speculate that it may also result from direct stimulation by factors released from dysplastic hepatocytes.
Myxoglobulosis, a rare variant of appendiceal mucocele, occurring secondary to an occlusive membrane.
A 77-year-old man presented with altered bowel habit and episodic right lower abdominal discomfort. A barium enema showed a large smooth filling defect in the caecum and at operation his appendix was seen to be hugely distended. Pathological examination showed myxoglobulosis, a rare variant of mucocele of the appendix. The appendiceal orifice was completely occluded by an imperforate membrane with no communication between the appendix and caecum. This is the first published report in the English language of appendiceal myxoglobulosis in association with such an occlusive membrane.
Association between risk of seroconversion of sentinel cattle to bluetongue viruses and Culicoides species (Diptera: Ceratopogonidae) in Queensland, Australia.
The association between risk of seroconversion of sentinel cattle to bluetongue viruses and the number of Culicoides brevitarsis Kieffer and C. wadai Kitaoka caught by light traps was investigated using survival analysis. Eight sentinel herds that seroconverted to bluetongue viruses between 1990 and 1994, and for which insect-trapping data were available, were selected for inclusion in the study. These herds were located at six sites along the eastern coast of Queensland, Australia, from approximately latitude 10 degrees South to 25 degrees South. C. brevitarsis was detected at all locations where sentinel herds were maintained, whereas C. wadai was detected at only two locations in northern Queensland where four sentinel herds were maintained during the study period. The mean number of C. brevitarsis and C. wadai caught per month was 230 and 21, respectively. A significant (P = 0.05) positive association was found between the risk of seroconversion of sentinel cattle to bluetongue viruses and the number of C. wadai caught in the same month.
PCR ribotyping and endonuclease subtyping in the epidemiology of Burkholderia cepacia infection.
Because of conflicting data about hospital-based transmission of Burkholderia (Pseudomonas) cepacia, an important respiratory pathogen in cystic fibrosis (CF), we compared strains found in sputum, lung, or blood of 29 CF patients in our center from 1988 to 1994, studying the relationship between strain and hospital exposure of incident and that of prevalent cases. Exposure was defined as a concurrent hospital stay between a prevalent and an incident case. B. cepacia strains were determined by polymerase chain reaction (PCR) ribotyping and endonuclease subtyping. The 16S to 23S spacer regions of the bacterial ribosomal RNA (rRNA) genes were amplified by PCR, and the product-size patterns used to type each B. cepacia isolate. Endonuclease digestion of the PCR products provided length polymorphisms for subtyping. There were 17 incident events during the period from 1988 to 1994, 16 of which involved a single ribotype. These 16 ribotypes could be divided into five subtypes by endonuclease mapping. Four patients grew B. cepacia from the blood, with the organism being the same strain as found in the lung in each case. Case controls were obtained to evaluate risk factors for B. cepacia acquisition. Concurrent hospitalization with a prevalent case significantly increased the risk of acquisition. There was no association between length of hospitalization, length of exposure, or FEV1 and the risk of B. cepacia acquisition.
Spatial analysis of seroconversion of sentinel cattle to bluetongue viruses in Queensland.
OBJECTIVE: To assess quantitatively the spatial distribution of seroconversion of Queensland cattle to bluetongue viruses. DESIGN: A sentinel herd study. SAMPLE POPULATION: Sixty-nine sentinel herds at 30 locations. PROCEDURE: Spatial clustering of seroconversion to bluetongue viruses was investigated during the period from 1990 to 1994. RESULTS: Seroconversion to only two bluetongue virus serotypes, 1 and 21, was observed. The 14 herds, in which seroconversion to bluetongue virus serotype 1 was detected, were located only along the eastern coastal and subcoastal region of Queensland, and were significantly (P < 0.05) clustered. Locations at which seroconversion to serotype 21 was detected, were not significantly clustered. The results generally agree with field observations, except for the failure to detect seroconversion to bluetongue viruses in north-western Queensland. CONCLUSION: Bluetongue infection of cattle in north-western Queensland may be temporally sporadic. The dominance of serotype 1 in the Queensland cattle population may be the result of differential transmission by potential vector species. Long-term surveillance programs are important for defining disease status of animal populations.
Hepatitis C virus: epidemiology and genotypes in the north east of England.
The epidemiology of hepatitis C virus (HCV) infection was studied in an English teaching hospital over an 18 month period. A total of 104 HCV antibody positive patients were referred for further investigation. They were divided into those diagnosed through screening (blood donors and intravenous drug abusers) and those diagnosed for other reasons, and their mean ages, known risk factors for HCV transmission, genotypes, and liver biopsy histology were analysed. Screened patients were significantly younger than the others. No significant difference in age was found between genotypes. Most patients genotyped (69%) were genotype 1. Intravenous drug abusers had a higher proportion of subtype 1a, and patients who acquired HCV through blood transfusion had a higher proportion of subtype 1b. Liver biopsy specimens were scored using a histological activity index for liver inflammation and fibrosis. Patients with subtype 1b had significantly more severe liver disease than other genotypes when the histological activity index scores for fibrosis were analysed (p < 0.05). Liver disease worsened significantly with age according to all three histological activity index scores (portal activity: p < 0.01, acinar activity: p < 0.001, fibrosis: p < 0.0001). Liver disease worsened with increased duration of infection (p < 0.002), and patients who also abused alcohol presented at a significantly younger age (cirrhosis, p < 0.05, hepatocellular carcinoma, p < 0.02).
Evaluating physical functional outcomes: one category of the NOC system.
All professional health care disciplines are now being held accountable to demonstrate the outcomes they produce. It is imperative that nurses demonstrate the difference they make in patient care to justify the cost of professional nursing. Standardized language is needed to aggregate data and report information on nursing interventions and associated outcomes. The Nursing Outcomes Classification (NOC) system is the first comprehensive classification of nursing-sensitive patient outcomes. In this review, the development of this classification system with specific results related to physical functional status and implications of NOC for nursing practice are discussed.
Infection of cattle with bluetongue viruses in Queensland, Australia: results of a sentinel herd study, 1990-1992.
Between 1990 and 1992, 47 sentinel herds of 10-20 cattle each were established throughout Queensland, Australia to monitor bluetongue virus infection. Sixteen herds at 12 locations seroconverted to bluetongue viruses during the study. Herd incidence rates ranged from 0.0 to 3.45 seroconversions per cattle-year at risk. The mean incidence rate was 0.29 seroconversions per cattle-year at risk (95% confidence interval 0.23-0.36), and the median incidence rate was 0.32 seroconversions per cattle-year at risk (95% CI 0.0-0.54). Only seroconversions to bluetongue virus serotypes 1 and/or 21 were detected in the study; no evidence was found of infection with other serotyped previously isolated in Australia (3, 9, 15, 16, 20, 23). Seroconversion to serotype 1 occurred more frequently than seroconversion to serotype 21. Seroconversion generally occurred between April and July, being most common in May, but varied considerably within each of the 3 years of the study. This study confirmed the presence of only serotypes 1 and 21 in Queensland and demonstrated the sporadic and restricted nature of bluetongue virus infection in the Queensland cattle herd.
Immunohistochemical detection of p53 protein expression in mammary carcinoma: a study of 80 cases.
Overexpression of p53 protein appears to be a common event in primary mammary carcinoma. This study was performed on 80 primary carcinomas of varying type and grade. Tumor p53 protein expression was detected by means of immunohistochemistry using the monoclonal antibody D07 on formalin fixed paraffin-embedded tissue sections. p53 overexpression was found in 50 (63%) of the 80 tumors. The correlation between p53 expression and high tumor grade, presence of in situ component and lymphocytic response was significant. However, no significant association was found between p53 expression and tumor size, age of the patients or tumor vascular invasion. Our study suggested that the immunohistochemical assay was a simple method of detection of mutant p53 proteins in routinely processed paraffin-embedded tissues of primary mammary carcinoma.
An assessment of partial rescreening as an internal quality control method for cervical smears.
Partial screening was performed on 10,800 cervical smears, comprising 8640 filed negative and unsatisfactory smears and 2160 newly received smears prior to conventional screening. Each slide was screened for 30 s and those considered abnormal were reviewed by standard screening. Partial screening led to the detection of 27 additional infections and 44 additional cytological abnormalities. These detection rates are better than those obtained with the traditional method of rescreening only a proportion of smears. Amongst the smears partially screened before conventional screening, partial screening detected 37-66% of infections and 22-71% of cytological abnormalities. We recommend the use of partial rescreening of all negatively reported smears as a method of internal quality control in cervical cytology laboratories.
The effects of vaccination of Merino ewes with an attenuated Australian bluetongue virus serotype 23 at different stages of gestation.
A cell culture attenuated Australian bluetongue virus serotype 23 (BLU23) prototype vaccine was assessed for its effects on pregnant Merino sheep. Seventy-six ewes were vaccinated at 5 different stages of gestation, and the failure to lamb at term was as follows: 35 to 43 days of gestation, 20/36 (56%); 57 to 64 days of gestation, 3/10 (30%); 81 to 88 days of gestation, 3/10 (30%); 109 to 116 days of gestation, 0/10 (0%); 130 to 137 days of gestation, 0/10 (0%). Of 30 ewes vaccinated with a cell culture supernatant fluid control between 35 and 43 days of gestation, 6.7% (2/30) failed to lamb at term. Two ewes vaccinated with BLU23 vaccine between 35 and 43 days of gestation had lambs with hydranencephaly. All other lambs born were clinically normal. Three ewes vaccinated with BLU23 aborted. Two of these were vaccinated between 35 and 43 days of gestation, the 3rd between 81 and 88 days of gestation. Five lambs were born with BLU group antibody. Four of these were from ewes vaccinated between 35 and 43 days of gestation, and 2 of these had hydranencephaly. The fifth was from a ewe vaccinated between 57 and 64 days of gestation. The vaccine did not produce disease in adult sheep, but was a potent cause of early foetal death and to a much lesser extent foetal malformation.
G protein gene mutations in patients with multiple endocrinopathies.
Point mutations of G protein genes that result in the constitutive activation of G proteins have been described. Such mutations have been shown to occur in a number of endocrine diseases. We have examined tissues from patients having more than one organ affected by an endocrine disorder and patients having separate distinct endocrine diseases for G protein gene mutations. G protein genes encoding for Gs alpha and Gi2 alpha were examined for activating mutations at codons 201 and 227 (Gs alpha) and codons 179 and 205 (Gi2 alpha) using site-directed oligonucleotide hybridization and direct sequencing of tissue DNA amplified by polymerase chain reaction. Tissues from six patients were examined. The only mutation that was identified was at codon 201 of Gs alpha (gsp), which encoded a change from arginine to cysteine. Patient 1 had the mutation in a corticotroph adenoma, a chemodectoma, and a nodular hyperplastic adrenal gland. patient 2 had the mutation in an extraadrenal pheochromocytoma, but an adrenal gland with medullary hyperplasia was wild-type. Patient 3 had an aggressive corticotroph adenoma and developed Nelson's syndrome after bilateral adrenalectomy. The corticotroph adenoma was wild-type, but both hyperplastic adrenal glands had the mutation. Patient 4 had the mutation in a parathyroid adenoma and in two hyperplastic parathyroid glands. Patient 5 had the mutation in both a primary and a metastatic pheochromocytoma. Patient 6 had the mutation in a parathyroid adenoma and also in histologically normal thyroid and parathyroid tissue. Leukocyte DNA was examined from five patients and was found to be wild-type in all cases. We conclude that G protein gene mutations occur in a wider range of endocrine conditions than has been recognized hereto. In addition, the presence of gsp mutations in different endocrine disorders in the same patient is suggestive of a common underlying etiology.
The discovery and structure-activity relationships of 1,2,3,6-tetrahydro-4-phenyl-1-[(arylcyclohexenyl)alkyl]pyridines. Dopamine autoreceptor agonists and potential antipsychotic agents.
A novel dopamine (DA) autoreceptor agonist, 1,2,3,6-tetrahydro-4-phenyl-1- [(3-phenyl-3-cyclohexen-1-yl)methyl]pyridine (14), was identified. The structure-activity relationships surrounding this compound were studied by synthesis of analogues and evaluation of their dopaminergic activity. The cyclohexene substitution pattern was varied along with the length of the chain connecting the 1,2,3,6-tetrahydro-4-phenylpyridine to the cyclohexene. Compound 14, having the 1,3-substitution pattern and a single methylene chain, was the most potent. The 1,2,3,6-tetrahydro-4-phenylpyridine could be replaced by other aryl-cyclic amines with a slight loss in activity. The phenyl group on the cyclohexene ring could be para substituted; electron-donating groups were better tolerated than electron-withdrawing groups. Finally, the enantiomers of 14 were resolved via the 1,1'-binaphthyl-2,2'-diyl hydrogen phosphate salts. Although both isomers were partial DA agonists, the (+)-enantiomer had higher intrinsic activity than the (-)-enantiomer. Syntheses were developed that allowed rapid preparation of analogues. An X-ray crystal structure determination of an intermediate identified the (+)-isomer of 14 as having R configuration. This compound, designated CI-1007 (PD 143188), was found to have antipsychotic-like activity in behavioral tests; in particular, it was orally active in the conditioned avoidance test in squirrel monkeys with an ED50 of 0.6 mg/kg. The overall profile suggests that (R)-(+)-14 may be a clinically useful antipsychotic agent.
Chronic peripheral nerve pain treated with direct electrical nerve stimulation.
Chronic somatic peripheral nerve pain was treated prospectively in 24 nonrandomized patients by a program of direct electrical nerve stimulation. Patients qualified for the program if anesthetic (lidocaine) nerve block of the involved cutaneous zone of the peripheral nerve relieved symptoms and transcutaneous electrical nerve stimulation transiently improved and did not exacerbate somatic pain. Results were judged according to a pain score. Patients noted improved sleep and complete absence of the need for narcotic pain medication. On the basis of subjective and objective criteria, 18 patients had good or excellent results and 6 had implant failures. Of the six patients with failures, three failed the trial period and did not have implantation, and three had no significant pain relief and were judged as treatment failures. Three patients had late equipment failure after initial good results. Most patients had some relief of pain, which increased their quality of life and eliminated the need for narcotic analgesia. Direct electrical nerve stimulation should be considered for somatic peripheral nerve pain that has not been ameliorated with other methods. It will reduce, although not necessarily eliminate, pain and pain behavior in most patients.
Infection of Culicoides brevitarsis and C. wadai (Diptera: Ceratopogonidae) with four Australian serotypes of bluetongue virus.
Field collected Culicoides brevitarsis Kieffer and C. wadai Kitaoka were fed on sheep that had been artificially infected with a field-isolate of either bluetongue virus serotype 3 (BLU3), BLU9, BLU16, or BLU23. Feeding rates averaged 11.9% but were variable. Survival of midges during incubation tended to be enhanced by the addition of antibiotics and fungicide to the diet. Attempts to transmit virus to sheep by the bite of these two species were unsuccessful. Both C. brevitarsis and C. wadai had infection rates of between 1.3 and 1.5% for BLU16 and BLU23. The infection rate for BLU3 in C. wadai was low at 0.07% whereas those for BLU3 in C. brevitarsis and BLU9 in both species could not be calculated because of insufficient virus isolations. The relatively high infection rates for BLU16 and BLU23 suggest that both C. brevitarsis and C. wadai are capable of circulating these viruses in the field.
Quantitative analysis of macrophages and perisinusoidal cells in primary biliary cirrhosis.
Immunohistochemistry and image analysis were used to quantify alterations in the Kupffer cell and 'activated' perisinusoidal cell populations in the different stages of primary biliary cirrhosis. Anti-CD68 macrophage antibodies were used to detect Kupffer cells, and anti-alpha-smooth muscle actin (alpha-SMA), PR 2D3 and anti-desmin antibodies to detect perisinusoidal cells. Liver biopsy material was available from 26 patients with primary biliary cirrhosis and 23 patients with histologically normal liver. Increased Kupffer cell numbers were observed in periportal/periseptal zones of stage 3 primary biliary cirrhosis (n = 9), and in random parenchymal areas of stage 3 and stage 4 cases. Significantly increased 'activated' perisinusoidal cell numbers were seen only in periportal/periseptal zones of stage 3 and stage 4 primary biliary cirrhosis. Neither Kupffer cell nor perisinusoidal cell numbers altered significantly in stage 1 and 2 primary biliary cirrhosis (n = 6). PR 2D3 positivity and increased alpha-SMA immunoreactivity by perisinusoidal cells in primary biliary cirrhosis support myofibroblastic differentiation of these cells. Human perisinusoidal cells, unlike their rodent counterparts, did not express desmin in primary biliary cirrhosis or control liver. Kupffer cells and 'activated' perisinusoidal cell accumulation in periportal/periseptal zones of the precirrhotic and cirrhotic primary biliary cirrhosis liver support the concept of Kupffer cell-mediated stimulation of perisinusoidal cells. Furthermore, these findings indicate that Kupffer cell-perisinusoidal interactions play an important role in the development of liver fibrosis and cirrhosis in primary biliary cirrhosis.