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BRITISH,AMERICAN TOBACCO COMPANY LIM|TF~D AUTHORISATION TO WORK WITH UNFENCED MACHINERY/LIVE EOUIPMENT_ Under the terms of the BATCo Technology Centre Standing Instructions and the Operations at Unfenced Machinery Regulations 1938, this letter formally confirms that your training and experience qualifies you as an 'authorised person' to work with unfenced machineD' or live equipment, as qualified below. In the case of electrical circuitry you should operate to the requirements of Electricity at Work Regulations SI 1989 No. 635 (see attached extract). It must be emphasised that such working practices should only be adopted when completely unavoidable. Whenever possible you must ensure that another person is working with you and if this is not in any way possible your intended manner of work should be agreed with your supervisor. You should be aware that many accidents occur when cleaning or inspecting machinery without undertaking proper safety procedures. This does not mean that because of your 'authofised' status you should abuse the position to carry out inspections without making electrical or mechanical systems safe. At all times you have a fight to discuss with your supervisor how to do something you believe to be dangerous. You also have an absolute duty to avoid endangering yourself or others in the way you work. To assist in the control of such work, you are to agree with your supervisor a system for recording all work covered by this authorisation, in a permanent record book. Where work can be agreed as "minor" this may be retrospective. Subje~ to the foregoing N. D. Warren is authorised to work with live instruments/computer circuitry Date of issue: 17 January 1994 Signature of Manager ........ ~.~ ........... V~ii,! until 3 lst~ 199_~5 Signature of holdef~..bC?...~"-'77- .~ ............... "~i .............. Footnote: 2 copies - one to be held on file one to be held by authofised person. Version: 14th January 1993 AQ9247

ELECTRICITY AT WORK - REGULATIONS LAID IN PARLIAMENT Some of the most far-reaching regulations produced by HSC since 1974 were laid in Parliament by Mr. Norman Fowler, Secretary of State for Employment, on 25 April. The Electricity at Work Regulations 1989 replace a range of inflexible and outdated. existing legislation with a comprehensive.and systematic approach to the control of electrical hazards covering all types of work and workplace. The new regulations • codify good practice as used now by responsible and well-informed employers, and will come into effect on 1" April 1990. Dr. Iohn Cullen, Chairman ofllSC, said "The new regulations, Code of Practic~ and Guidance replace regulations which are over 80 years old and which had limited applicabili .ty. They are the culmination of many years' painstaking work by the Commission and Executive in discussion, formulation, negotiation and drafting. A key favor in the regulations is a requirement for employers to adopt a 'switch-off- list' approach to work on electrical equipment. Live working which at present accounts for a substantial proportion of electrical accidents, will, under these regulations, be permitted only it'it is unreasonable in all the circumstances for the system to be dead; it is reasonable in all the circumstances for the work :to be carried out live; and suitable precautions are taken to prevent injury". Elecricity at work regulations, SI 1989 No. 635, ISBN 0 11 096635 X,, £2.65 from t-]MSO or booksellers. AQ9247

I Comparative Analysis of F.nvironmentai llaza-~ds from Professor .lames M. Parry, School of Riological Sciences, UnJver~ify College of Swansea SA2 8PP. A variety of environmental expo~ure,~ such as environmental tobacco smoke (~TS), dlese[ exhaust particulates and 8en~ral ~tmo~pheric pollution has been implicated in the etJoJosy of vnrinus health problem~ ~uch as c~ncer~ of the respiratory tract. In ~ome case~, epi~emiolo~Jcal ~B~B ha~ been used to postulate various quantita~ve risks associ.ated wilh these exposures. However, as with most epJdemEoloEica] studie~ the prohlem~ of confn~ndin8 fa~tnr~, particularly ~Jth reference to ~TS, has resulted in the risk estimates be~n8 Jm p rec~se. The aim o~ the proposed project is to d#velop methodolo~ie~ which will ~o~ ¢omparJson~ to be made o( varEo.s environmental exposure~. these comparisons would be performed usJnB laboratory technique~ to determine the e~fects of environmental samples upon the Jnductlon of a variety oY ~ene~c and other bioloEic~l endpoints. EndpoJnts Tobacco particulate matter (TPM) collected updn CambrJdS~ filters be used to ~enerate solvent extr~et~ which produce a vmr~ety nf ~ff~nt~ appropriate cuJtured cell ~y~tem~. These effects include:- l) the induction of DNA ]Psion~, ~s measured by the use o( the po~tlahellinK teehniquPs, the induction of point mur~tinn~, 3} the induction of chrnmo~nmP ~tru~tur~l ~nd numPri~al damage, the induction of aberrations of mitotic cell d[vJsJ.on and modifications cell proliferation and toxi~i?y, the indt~c~on of mRlignant c~ll tr~n~formatlhn. In v~P~ of the complex mix of potentially genotoxie ~h~mic~l~ fmmd in AQ9247

2 TPM the use of appropriate metabolic activation systems is an essential requirement for the analysis TPM extracts. For the purpose of our proposed investigation it is our intension to make use of genetically engineered cell lines which express a variety of enzymes such as cytochrome P-~50 |soenzyme~ implicated in the activation of various pro-mutagens. Solvent extracts or pure samples of each environmental sample wil~ be used to treat genetically engineered cell I/nes and a variety of endpoints quaniJfied. These endpoints have been selected on the basis of their ability to predict the initiation and promoting capacity of biologically active materials. The endpoints will be evaluated in the following order and as illustrated in Figu re I. a) The frequency of induced DNA adducts wit[ be determined using the post-labelling technique. Note, in these studies we will employ a standard of DNA treated with the mutagen ~-nltroquinaline oxide to ensure both the quality of the ~2p post-labetling assays and their quantitation. This assay will provide data providing information on the frequency of lesions which provide the starting point for the induction of mutations. b) The frequency of induced micronuciel in bJnucleate cells produced by treatment with the actin inhibitor cytochalasin B. The induction of micronuclei provides a measure of the induction of chromosomal damage leading to structural chromosome changes and mod|ficatJons of chromosome segregation Ieadlng to numerical chromosome changes. Treatment of mJcronuclei with centromere specific DNA probes aUows the classification of mlcronuclei into those resulting from either chromosome structural or numerica/ changes. c) The frequency of point mutations will be determined by the u.~e of the Restriction Site Mutation Assay (RSM). We have developed the assay to AQ9247

3 measure the induction of mutations in the DNA ,~c~nces coding for bacterial restriction enzyme recognit(on sites. The methodology will be used to quantify the induction of rnl~tatinn.~ in th~ spqttenc.e of the P53 turno~Jr suppressor gene~ which has been extensively u.~ed to determine the profile of mutation induction Jn turnouts following expo.~ure to environmental genotoxins. d) The frequency of induction o£ rnodificatlons of the fidelity of the synthesis and functioning of the mitotic spindle and ceil division. This endpolnt provides a valuable rneasure of the induction of disturbances to cell division including toxicity and ,~uh,~eqttent rni.~segregatlnn chromosomes leading to numerically abnormal progeny e) The frequency of induction of malignant cell transformation in Syrian hamster dermal cells. In this endpoint, cells are exposed to a ~JngJe Jn|tJating dose of a chemical genotoxJn and then exposured to graded doses of the test agent ~or four weeks to assess the ability of treatment to ~promote" the induction of cell transfformatlon. It should he noted that this endpolnt requires a longer time sca~e than those, outlined in a) to d) and tt ls our intention to use this methodology with a lower priority. The decl.~ton to utilize the transformation endpoint would he based upon the results obta/ned Jn assays a) to d). The package o~ assays outlined above would provide a lahnratory based method for comparing the. quantitative ha~.ard,~ of various environmental exposures, l:f .~uccessful the basic methodologies could hp applied to samples such as buccal smears Jn humans e×posed to a variety environmental Sample Collection The overall a~m of the project is to develop methodologie~ to compare the relative hazards of various environment exposure~. The intended AQ9247

exposure~ include benT.ene, pesticides and agents ~uch'~as environmental tobacco smoke (iTS). It is our initial intension to focus our activity upon particulate matter derived from various sources generated in the laboratory and in environmental situations. At this stage we would appreciate advic.e on the methods to be used to collect environmental samples particularly with regard to iTS. Samples wig be collected by pumping air through Cambridge filters for varying periods of time, dependent upon the nature and lev~.| of the environmental exposure. Environmental samples will be collected using battery powered oumps via Cambridge Filters. 1) TPM will be collected from low tar cigarettes onto Cambrldge filters and will be extracted with a suitable solvent such as dimethy[ s,~lpboxide. 2) Particulate matter will he collected from the environmental smoke produced in a confined chamber following the smoking of a specific number of cigarettes. 3) Following an agreement made with the loc~] LJcenced Victual.lets Association, particulate matter will be collected from a number of public bars. Sampling times and TPM collected will be calibrated by counting cigarette ~tubs collected over a fixed period of time. #) Particulate matter will be collected from a "smoking allowed" area for a fixed period of time and correlated with cigarette st,~b~ collected. Particulate matter will be c~llected from a "non-smoking" area. 6) Particulate matter samples will be collected from the exhaust of a small car engine running at a variety of speeds. 7) Particulate matter samples wiJJ be collected from the exhaust of a "heavy load" di~.sei engine. 8) Particulate matter will he collected from a city centre, ~ rural area and a bus maintenance garage. Particulate matter collected as described above will be solvent extracted AQ9247

5 (preferably DMSO) and evaluated for biological activity as ol~l£ned above. The activity of the particulate extracts will be compared to the activity in the same test systems of a number of other environmental toxins including benzene and pesticides including the rodenticJde c~lcium phosph~mi~le. These latter aaents are open to discussion and may be modified and expanded. CostJn ~ s The project outlined could most efficlentiy be performed over a period of 3 years by a full t~me post-doctoral fel|ow and a half time technician. The post doctoral worker would be responsible for aH the biological sl.ud|es involving the evaluation of the genotoxic activity of the extracts and the technician will be responsible for sampl~ collection and preparation. An alternative would be to inJtiate the study with a post doctoral, worker for a period of 12 months and following system calibration to continue the study with a post-graduate student. Costings are provided )~or one year to provide an indication of overall financial implicationS. I) Salary Costs Full Post doctoral Re.~earcher £18,000 2) Salary Costs half-time technician £ 7,500 3) Superannuation and NatJonP.l In.~urance £ f;,375 on !) and 2) 25% of Salary costs t~) Consumable Costs £ 8,500 5) Equipment 0 6) Sub total £~0,375 7) University Overheads ~0~ of Sub total £16,750 AQ9247

FiF, u re I Outline of procedure to he used to evnhJat~ the bioJo~Ecnl a) b~ d~ activity of solvent extracts Solvent extracts Cultured human cell lines expre.~sing activating enzymes Continuous exposure for up to two ceil cycles Samples evaluated for: Frequency of r)NA adducts, Frequency of induced mJcronucle| classified for the presence and absence of centromere DNA~ Frequency of induced mutations in the P53 turnout suppressor gene, Frequency of induced aberrations of cell division. Solvent extracts Corttinuous exposure of Syrian hamster dermal cells for ~ weeks following exposure to a ,~ingJe initiating dose of a chemicaJ genotoxJn. Assessment of the induction of malignant transformation AQ9247

Sam pie Collection I) ~) 7) 9) 1o) TPM collected from low tar cigarette, Particulate matter collected from ETS under laboratory conrliti~n.% Partim~late matter collected from ETS present in a bar of a pl~blic house Particulate matter collected from ETS present in a "smoking a]lowed" area of the University, Particulate matter collected from a "non-smoking" area, Particulate matter coUected from the exhaust of a small car diesel engine, Particulate matter collected from the exhaust of a "heavy load" die,~el engine, Particulate matter collected from a city centre area I]tilized by a mixture of traffic, Particulate matter collected from a floral area. Particulate matter collected from a bus maintenance garaEP, Chemical samples of envJronmentPJ e×po,~l~res including benz~ne and a ranKe of pesticides including the rodenticide calcium phosphamld~. A(~9247

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