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Biomedical subjects

J C Simpson

Publications and source records attributed to J C Simpson.

At least 19 recordsLinked to original sources

Ricin A chain utilises the endoplasmic reticulum-associated protein degradation pathway to enter the cytosol of yeast.

Cytotoxic proteins such as ricin A chain (RTA) have target substrates in the cytosol and therefore have to reach this cellular compartment in order to act. RTA is thought to translocate into the cytosol from the lumen of the endoplasmic reticulum (ER), although how it traverses the ER membrane has not been established. Using yeast mutants defective in various aspects of the ER-associated protein degradation (ERAD) pathway, we show that RTA introduced into the yeast ER subverts this pathway to enter the cytosol via the Sec61p translocon. A significant proportion of the exported RTA avoided proteasomal degradation. These data are consistent with the contention that the RTA component from ricin endocytosed by mammalian cells may likewise exploit ERAD to translocate into the cytosol.

Biological Transport

Evidence for a COP-I-independent transport route from the Golgi complex to the endoplasmic reticulum.

The cytosolic coat-protein complex COP-I interacts with cytoplasmic 'retrieval' signals present in membrane proteins that cycle between the endoplasmic reticulum (ER) and the Golgi complex, and is required for both anterograde and retrograde transport in the secretory pathway. Here we study the role of COP-I in Golgi-to-ER transport of several distinct marker molecules. Microinjection of anti-COP-I antibodies inhibits retrieval of the lectin-like molecule ERGIC-53 and of the KDEL receptor from the Golgi to the ER. Transport to the ER of protein toxins, which contain a sequence that is recognized by the KDEL receptor, is also inhibited. In contrast, microinjection of anti-COP-I antibodies or expression of a GTP-restricted Arf-1 mutant does not interfere with Golgi-to-ER transport of Shiga toxin/Shiga-like toxin-1 or with the apparent recycling to the ER of Golgi-resident glycosylation enzymes. Overexpression of a GDP-restricted mutant of Rab6 blocks transport to the ER of Shiga toxin/Shiga-like toxin-1 and glycosylation enzymes, but not of ERGIC-53, the KDEL receptor or KDEL-containing toxins. These data indicate the existence of at least two distinct pathways for Golgi-to-ER transport, one COP-I dependent and the other COP-I independent. The COP-I-independent pathway is specifically regulated by Rab6 and is used by Golgi glycosylation enzymes and Shiga toxin/Shiga-like toxin-1.

ADP-Ribosylation Factor 1

Evaluating Tackling Rugby Injury: the pilot phase for monitoring injury.

OBJECTIVE: To assess the suitability of two previously unused data sources for monitoring rugby injury throughout New Zealand. METHOD: Interviews were conducted with respondents sampled from players registered with the Rugby Football Unions (RFUs) and players claiming for rugby injuries from the Accident Rehabilitation and Compensation Insurance Corporation (ACC) in Auckland and Dunedin. RESULTS: Of the 500 RFU players sampled, 63% were interviewed and of these 39 (12%) had been injured playing rugby union. Of the 456 ACC claimants sampled, 66% were interviewed and 265 (88%) had been injured playing rugby union. CONCLUSION: Identifying injured players through ACC claims was more efficient, both procedurally and because a smaller sample size was required to detect changes in incidence. IMPLICATIONS: With no routine surveillance of sports injury being undertaken, recording sporting codes in national injury surveillance systems would assist the monitoring of sports injury.

Adolescent

The KDEL retrieval system is exploited by Pseudomonas exotoxin A, but not by Shiga-like toxin-1, during retrograde transport from the Golgi complex to the endoplasmic reticulum.

To investigate the role of the KDEL receptor in the retrieval of protein toxins to the mammalian cell endoplasmic reticulum (ER), lysozyme variants containing AARL or KDEL C-terminal tags, or the human KDEL receptor, have been expressed in toxin-treated COS 7 and HeLa cells. Expression of the lysozyme variants and the KDEL receptor was confirmed by immunofluorescence. When such cells were challenged with diphtheria toxin (DT) or Escherichia coli Shiga-like toxin 1 (SLT-1), there was no observable difference in their sensitivities as compared to cells which did not express these exogenous proteins. By contrast, the cytotoxicity of Pseudomonas exotoxin A (PE) is reduced by expressing lysozyme-KDEL, which causes a redistribution of the KDEL receptor from the Golgi complex to the ER, and cells are sensitised to this toxin when they express additional KDEL receptors. These data suggest that, in contrast to SLT-1, PE can exploit the KDEL receptor in order to reach the ER lumen where it is believed that membrane transfer to the cytosol occurs. This contention was confirmed by microinjecting into Vero cells antibodies raised against the cytoplasmically exposed tail of the KDEL receptor. Immunofluorescence confirmed that these antibodies prevented the retrograde transport of the KDEL receptor from the Golgi complex to the ER, and this in turn reduced the cytotoxicity of PE, but not that of SLT-1, to these cells.

ADP Ribose Transferases

Comparative personal exposures to organic dusts and endotoxin.

The aims of the study were to provide valid comparative data for personal exposures to dust and endotoxins for different occupations and to calculate comparative data for the contamination of organic dusts with endotoxin. Nine different occupational settings were studied, drawn from the textile, agricultural and animal handling industries. Samples were collected by personal sampling techniques, using the Institute of Occupational Medicine (IOM) sampling head, glass fibre filters and rechargeable sampling pumps. The dust exposures were calculated by gravimetric analysis and using the calculated volume of air sampled were expressed as mg/m3. Endotoxin exposures were measured using a simple water extraction from the collected dusts, followed by a quantitative turbidimetric assay. Results were expressed as ng/m3, using the calculated volume of air sampled. In addition, the levels of the contamination of dusts with endotoxin for individual industries were expressed as ng/mg of collected dust. Two hundred and fifty-nine samples, collected from 9 different industries and across 36 different sites were analysed. This represented a sampling rate of 25% for the total work force. The average sampling time was 4.62 h. For all the dusts collected, a significant correlation between the collected dust and endotoxin was seen (r = 0.7 and p < 0.001). The highest dust exposures occurred during cleaning activities (grain handling: 72.5 mg/m3). The individuals exposed to the highest median level of dust and endotoxin were the animal handlers (poultry handlers, dust: 11.53 mg/m3, endotoxin: 71,995 ng/m3). Weaving and mushroom cultivation had the lowest exposures for dust and endotoxins. The mostly highly contaminated dusts (median values expressed as ng of endotoxin per mg of collected dust) were found in the animal handling (poultry: 1,030 ng/mg, swine: 152 ng/mg) and cotton spinning (522 ng/mg) industries. Processing of cotton and wool fibres was found to reduce the levels of contamination of dusts with endotoxin. In the study, valid comparative data for personal exposures to organic dusts and endotoxins have been presented. The highest exposures were found amongst animal handlers and during cleaning activities. The results highlight that dust exposures are greater in a number of industries than the set exposure standards. In addition, endotoxin exposures are found to be greater than levels at which harmful effects have been demonstrated.

Agriculture

Expression of mutant dynamin protects cells against diphtheria toxin but not against ricin.

Diphtheria toxin is believed to enter sensitive mammalian cells via receptor-mediated endocytosis from clathrin-coated pits, while ricin can enter via both clathrin-dependent and clathrin-independent endocytosis. The present study has confirmed this by determining the toxin sensitivity of COS-7y cells which were transiently overexpressing a trans dominant negative mutant of dynamin, a GTPase required for the budding of clathrin-coated vesicles from the plasma membrane. Cells overexpressing wild-type dynamin showed normal receptor-mediated endocytosis of transferrin and remained sensitive to both diphtheria toxin and ricin. Cells overexpressing a mutant dynamin defective in GTP binding and hydrolysis were unable to endocytose transferrin and were protected against diphtheria toxin, but they remained completely sensitive to ricin intoxication. Treating non-transfected cells or cells overexpressing mutant dynamin with nystatin caused a redistribution of the caveolae membrane marker protein VIP21-caveolin from the cell surface to intracellular locations, but did not affect their sensitivity to ricin. The redistribution of caveolin seen after nystatin treatment may reflect the disappearance of caveolae. If this is the case, caveolae are not responsible for the endocytosis of ricin. An alternative clathrin-independent route may operate for ricin, since cellular uptake, intracellular transport, and translocation into the cytosol remain unaffected when clathrin-dependent endocytosis is effectively blocked.

Animals

Educating future physicians for Ontario.

In 1987, Ontario's physicians conducted a strike, ultimately not successful, over the issue of "extra billing." The fact that the Ontario public did not support this action reflected a major gap between the profession's view of itself and the public's view of the profession. In 1990, the province's five medical schools launched a collaborative project to determine more specifically what the people of Ontario expect of their physicians, and how the programs that prepare future physicians should be changed in response. The authors report on the first five years of that ongoing project. Consumer groups were asked to state their views concerning the current roles of physicians, future trends that would affect these roles, changes in roles they wished to see, and suggestions for changes in medical education. Methods used included focus groups, key informant interviews, an extensive literature review, and surveys, including a survey of health professionals. Concurrently, inter-university working groups prepared tools and strategies for strengthening faculty development, assessing student performance, and preparing future leadership for Ontario's medical education system. Eight specific physician roles were identified: medical expert, communicator, collaborator, health advocate, learner, manager ("gatekeeper"), scholar, and "physician as person." Educational strategies to help medical students learn to assume these eight roles were then incorporated into the curricula of the five participating medical schools. The authors conclude that the project shows that it is feasible to learn specifically what society expects of its physicians, to integrate this knowledge into the process of medical education reform, and to implement major curriculum changes through a collaborative, multi-institutional consortium within a single geopolitical jurisdiction.

Consumer Behavior

Negative, positive, and disorganized symptom dimensions in schizophrenia, major depression, and bipolar disorder.

We compared the factor structure of positive and negative symptoms in schizophrenia (N = 214), major depression (N = 97), and bipolar disorder (N = 58) to determine whether schizophrenia factors would generalize to mood disorders. A study of schizophrenia and mood disorders identified patients whose symptoms were evaluated with the Scale for the Assessment of Negative Symptoms and the Scale for the Assessment of Positive Symptoms. We conducted principal component analyses with orthogonal rotation on the global ratings. The factor structure replicated earlier findings of three factors (negative, positive, and disorganization) in groups with schizophrenia, combined mood disorders, and psychotic symptoms, regardless of diagnosis. An additional negative symptom factor was found in the major depression group. The bipolar group did not have a disorganization factor. Similar symptom factors in schizophrenia and mood disorders suggest a continuity in the major affective and psychotic disorders that appears to reflect the underlying dimension of a psychotic process.

Adult

Prevalence and predictors of work related respiratory symptoms in workers exposed to organic dusts.

OBJECTIVES: The aims of this study were to document the prevalence of work related upper and lower respiratory tract symptoms in workers exposed to organic dusts and to identify variables predictive of their occurrence. METHODS: A cross sectional survey with an administered questionnaire (a previously validated adaptation of the Medical Research Council (MRC) respiratory questionnaire) was performed. Symptoms were classified as work related by their periodicity. Demographic data, smoking habits, and occupational histories were recorded. Personal exposures to dust and endotoxin were measured and individual subjects ascribed an exposure value specific to occupation, site and industry. Cox's regression techniques were used to identify variables predictive of work related upper and lower respiratory tract symptoms. Information was stored using Dbase 3 and analysed with SPSS. RESULTS: 1032 Workers (93% of the target population) were studied in nine different industries. The highest prevalences of work related lower respiratory tract symptoms (38.1%), upper respiratory tract symptoms (45.2%), and chronic bronchitis (15.5%) were found among poultry handlers. White workers were significantly more likely to complain of upper and lower respiratory tract symptoms. An individual in the swine confinement industry had a symptom complex compatible with byssinosis. Increasing current personal exposures to dust or endotoxin were found to be predictive of upper and lower respiratory tract symptoms, chronic bronchitis, and byssinosis. In a univariate analysis a relation between current exposures and the organic dust toxic syndrome was found. Present smoking and previously documented respiratory tract illness were significantly predictive of work related lower respiratory tract symptoms. Women were more likely to report work related upper respiratory tract symptoms. CONCLUSIONS: People exposed to organic dusts may have a high prevalence of work related respiratory tract symptoms which are related to dust exposures and smoking habits. Action should be taken to reduce exposures to dust and endotoxin and stopping smoking should be promoted among workers exposed to organic dusts to reduce morbidity.

Adult

Do radiographic features of acute infection influence management of lower respiratory tract infections in the community?

Community-acquired lower respiratory tract infection (LRTI) is a common clinical problem, and a chest radiograph is the most common investigation. This study investigated why general practitioners (GPs) request radiographs and whether radiographic features of LRTI affect management. The 12 month study included GP-requested chest radiographs performed at the Manchester Royal Infirmary. The GPs of patients with radiographic evidence of LRTI were sent a questionnaire. Radiographs (n=2,538) were performed in patients aged >15 yrs. One hundred and eleven (4.4%) chest radiographs showed evidence of LRTI, and 97 (87%) were included in the analysis. Fifty-six (62%) had their radiograph requested at the second or third visit. Forty-five (58%) radiographs were performed to confirm infection. In 47 (48%) of cases the GP felt the radiograph had affected treatment. An effect on treatment was significantly more common in patients not receiving antibiotics prior to the radiograph. Twenty-nine (32%) of GPs felt a normal radiograph would have affected management, most often with respect to antibiotic prescriptions (17 (58%)). Where the radiologist's report suggested a repeat radiograph or hospital referral, this was significantly more likely to have occurred. The commonest reason for a radiograph in patients with suspected lower respiratory tract infection is to confirm the infection. In the opinion of the general practitioner the results of the radiograph do affect patient treatment. The wording of the radiologists report is associated with patient management.

Acute Disease

Chronic bronchitis in textile workers.

BACKGROUND: Exposure to cotton is known to produce a specific occupational disease known as byssinosis. A large population of textile workers was investigated to determine whether such exposure was also associated with chronic bronchitis once other possible aetiological factors had been accounted for. METHODS: A total of 2991 workers were investigated for the presence of symptoms compatible with chronic bronchitis. An MRC adapted respiratory questionnaire and MRC definition of chronic bronchitis were used for diagnostic labelling. Current and lifetime exposure to dust was estimated by personal and work area sampling, and the use of records of retrospective dust levels previously measured over the preceding 10 years. Airborne endotoxin exposure was measured using a quantitative turbidometric assay. Lung function tests were performed to measure forced expiratory volume in one second (FEV1) and forced vital capacity (FVC). A control group of workers exposed to man-made fibre textiles was identified. The comparative prevalence of chronic bronchitis in the two populations was assessed, allowing for sex, age, smoking habit, and ethnic origin. Two case referent studies were also performed; cases of chronic bronchitis were separately matched with controls from the cotton and control populations to determine the effect of the symptomatic state on lung function. RESULTS: After controlling for smoking (pack years), workers in a cotton environment were significantly more likely to suffer from chronic bronchitis and this was most marked in workers over 45 years of age (odds ratio 2.51 (CI 1.3 to 4.9); p < 0.01). Regression analysis of all possible influencing parameters showed that cumulative exposure to cotton dust was significantly associated with chronic bronchitis after the effects of age, sex, smoking, and ethnic group were accounted for (p < 0.0005). In the intra-cotton population case control study a diagnosis of chronic bronchitis was associated with a small decrement in lung function compared with controls: percentage predicted FEV1 in cases 81.4% (95% CI 78.3 to 84.6), controls 86.7% (84.9 to 88.5); FVC in cases 89.9% (95% CI 87.0 to 92.9), controls 94.6% (92.8 to 96.4). After controlling for cumulative past exposure and pack years of smoking the effect of the diagnostic state remained significant for both FEV1 (p < 0.01) and FVC (p < 0.05). CONCLUSIONS: Chronic bronchitis is more prevalent in cotton workers than in those working with man-made fibre and exposure is additive to the effect of smoking. The diagnosis of chronic bronchitis is associated with a small but significant decrement in lung function.

Adult

Revisiting the factor structure for positive and negative symptoms: evidence from a large heterogeneous group of psychiatric patients.

OBJECTIVE: The factor structures of individual positive and negative symptoms as well as global ratings were examined in a diagnostically heterogeneous group of subjects. METHOD: Subjects were identified through a clinical and family study of patients with major psychoses at a VA medical center and evaluated with the Scale for the Assessment of Negative Symptoms and the Scale for the Assessment of Positive Symptoms. For the examination of global-level factor structures (N = 630), both principal-component analysis and factor analysis with orthogonal rotation were used. Factor analysis was used for the examination of item-level factor structures as well (N = 549). RESULTS: The principal-component analysis of global ratings revealed three factors: negative symptoms, positive symptoms, and disorganization. The factor analysis of global ratings revealed a negative symptom factor and a positive symptom factor. The item-level factor analysis revealed two negative symptom factors (diminished expression and disordered relating), two positive symptom factors (bizarre delusions and auditory hallucinations), and a disorganization factor. CONCLUSIONS: The generation of additional meaningful factors at the item level suggests that important information about symptoms is lost when only global ratings are viewed. Future work should explore clinical and pathological correlates of the more differentiated item-level symptom dimensions.

Adult

Testing for interaction between maternal smoking and TGFA genotype among oral cleft cases born in Maryland 1992-1996.

OBJECTIVE: Infants born in Maryland between June 1992 and June 1996 were used in a case-control study of nonsyndromic oral clefts to test for effects of maternal smoking and a polymorphic genetic marker at the transforming growth factor alpha (TGFA) locus, both of which have been reported to be risk factors for these common birth defects. DESIGN AND SETTING: Cases were infants with an oral cleft ascertained through three comprehensive treatment centers, with additional ascertainment through a registry of birth defects maintained by the Maryland Health Department. Controls were healthy infants. Medical history information on infants and mothers were collected, along with DNA samples. PATIENTS, PARTICIPANTS: Among 286 cases contacted (72% ascertainment), there were 192 nonsyndromic isolated oral clefts (106 M; 86 F) available for this case-control study. MAIN OUTCOME MEASURES: The largest group of 149 Caucasian nonsyndromic cases and 86 controls was used to test for association with maternal smoking and genotype at the Taq1 polymorphism in TGFA. RESULTS: While this modest sample had limited statistical power to detect gene-environment interaction, there was a significant marginal increase in risk of having an oral cleft if the mother smoked (odds ratio = 1.75, 95% CI = 1.01 to 3.02). We could not demonstrate statistical interaction between maternal smoking and TGFA genotype in this study, however, and the observed increase in the C2 allele among cases was not statistically significant. CONCLUSIONS: We could not confirm either the reported association between oral clefts and TGFA genotype or its interaction with maternal smoking. However, these data do show an increased risk if the mother smoked during pregnancy, and this effect was greatest among infants with a bilateral cleft and no close family history of clefts.

Alleles

Free ricin A chain reaches an early compartment of the secretory pathway before it enters the cytosol.

During the intoxication of mammalian cells by ricin, the catalytically active A chain must cross the membrane of an intracellular compartment in order to reach its ribosomal substrates in the cytosol. The actual site of ricin A chain translocation is unclear, and conflicting views hold that it enters the cytosol from endosomes or from an early compartment of the secretory pathway, possibly the lumen of the endoplasmic reticulum. Here we show that treating cells with brefeldin A, or transiently overexpressing mutant GTPases known to inhibit biochemical complexes mediating anterograde and retrograde transport between the endoplasmic reticulum and the Golgi complex, protected cells from intoxication by free ricin A chain. These data indicate that ricin A chain, either free or as part of intact ricin, reaches an early compartment of the secretory pathway before translocation into the cytosol occurs.

Amino Acid Sequence

Mortality among patients with schizophrenia.

Results from a 40-year followup study of psychotic patients are presented to illustrate some of the major findings and unanswered questions about excess mortality in schizophrenia and to suggest analytical approaches that take full account of the potential effects of sample heterogeneity. In this study, hospital-diagnosed schizophrenia patients were at increased mortality risk whether or not they met research criteria for schizophrenia. In addition, mortality outcomes of several major diagnostic groups were similar despite substantial clinical and demographic differences between the groups at baseline. These results suggest that both diagnosis-specific and nondiagnostic factors are needed to account for excess mortality in patients with major psychiatric disorders. The issue of heterogeneity is also crucial for the clinical purposes of predicting and ultimately reducing the mortality risk of psychiatric patients, for example, in delineating profiles of high-risk patients who are not necessarily typical of other patients with the same diagnosis.

Adult

Reconstruction of radionuclide releases from the Hanford Site, 1944-1972.

Historic releases of key radionuclides were estimated as a first step in determining the radiation doses that resulted from Hanford Site operations. The Hanford Site was built in southcentral Washington State during World War II to provide plutonium for the U.S. nuclear weapons program. As part of the Hanford Environmental Dose Reconstruction (HEDR) Project, releases to the Columbia River of 24Na, 32P, 46Sc, 51Cr, 56Mn, 65Zn, 72Ga, 76As, 90Y, 131I, 239Np, and nonvolatile gross beta activity from operation of eight Hanford single-pass production reactors were estimated. Releases of 90Sr, 103Ru, 106Ru, 131I, 144Ce, and 239Pu to the atmosphere from operation of chemical separation facilities were also estimated. These radionuclides and the atmospheric and Columbia River pathways were selected for study because scoping studies showed them to be the largest contributors to dose from Hanford operations. The highest doses resulted from releases to the atmosphere of 131I from chemical separations plants in the pre-1950 period. Prior to 1950, the technology for limiting iodine releases had not been developed. Hence, a very detailed reconstruction of the hourly 131I release history was achieved for 1944-1949 using Monte Carlo methods. Atmospheric releases of the other radionuclides were estimated on a monthly basis for 1944-1972 using deterministic calculations. Monthly releases to the Columbia River for 1944-1971 were based on Monte Carlo methods.

Air Pollutants, Radioactive

Point mutations in the hydrophobic C-terminal region of ricin A chain indicate that Pro250 plays a key role in membrane translocation.

A series of mutations have been made in the carboxyl terminus of ricin A chain, centred on the hydrophobic region between amino acid residues Val245 and Val256. The mutant ricin A chains were expressed to a high level in an Escherichia coli system and the proteins purified to homogeneity. The enzymic activity of each of these A chain molecules was tested on rabbit reticulocyte ribosomes; in all cases, the activities were found to be comparable to wild-type recombinant ricin A chain. Following reassociation of these A chains to ricin B chain, Vero cells were challenged with these holotoxins and the cytotoxicities determined. Mutant ricin A chain with Ile247-->Ala was unable to reassociate and form holotoxin, indicating the importance of this residue in the interaction with ricin B chain. Mutant ricin A chain with Pro250-->Ala readily reassociated with ricin B chain, forming holotoxin with a 170-fold reduction in cytotoxicity to Vero cells. Other mutations in this region also produced A chain proteins which gave marked reductions in holotoxin cytotoxicity. We propose therefore that the C-terminal hydrophobic region of ricin A chain may be involved in membrane interactions prior to the translocation of this subunit into the cytosol, and that Pro250 plays a key role in one or both of these steps.

Amino Acid Sequence