Showing papers in "Journal of Clinical Pathology in 1993"

Cowan and Steel's Manual for the Identification of Medical Bacteria

Abstract: The publication of the third edition of "Cowan and Steel," almost 20 years after the last, is like the return of a very dear, old friend. The beauty is that despite everything that has happened, the friend is at heart just the same-the mission "to help those who have isolated a bacterium (from a "medical" source) and want to identify it". The book begins with five general chapters on classification and nomenclature; culture media (with Frau Hesse still in her rightful place); isolation techniques; characterisation; and identification. I found the relative dismissal of the antibiogram as an aid to identification unsubtle, and would have liked to have more on the perils of over-reliance on kits. I believe, however, that the warning to the medical microbiologists of the future that the "materials and methods (that) are simply to be taken from the refrigerator or shelf as kits" may be a fool's paradise is timely. The essence of the book for the DIY enthusiast is to be found in chapters 6 and 7 in which all the major medical bacteria are considered in the familiar series of first, second, and third stage tables. Of course they are longer than before; there are, for example, now 26 staphylococcal species compared with eight last time. It seemed to me that a potential inconsistency which the authors challenge us to find concerns motility in Enterococcus faecium. On page 26 it is implied that many strains of E faecium are motile. Table 6.3b has Efaecium non-motile (up to 15% possibly motile according to the key to the tables), but E casseliflavus and E gallinarum motile. On page 66 they refer to E faecium "with its five varieties". What are the facts please? The authors continue a tradition of not being afraid to be different. They have not really given up much in relation to Kiebsiella (the little essay on the genus is great fun) and hang on to Acinetobacter iwoffii. Tables 7.5 and 7.6 are masterpieces of active speciation, being not quite up to date. However, they could have been a little more daring with the old Bacteroides genus, and Mobiluncus dosen't fare well. Overall, however, most of the organisms that turn up in a medical laboratory find a home here, even if only among such taxonomic evasions as "A group of difficult organisms". The Appendices are far from incidental and contain a great deal of valuable information on media and stains, characterisation tests and test organisms (all useful ammunition in current European discussions on standardisation), and on information processing. There are three sections on the bacteriological law in relation to taxonomy, and finally a useful glossary. There are almost 50 pages of references. This book is very definitely a must for anyone who calls him or herself a microbiologist, whether clinical, medical, or technological in persuasion. IAN PHILLIPS Quality Management fo: Safety, Environmental Hes No. 141. IPCS. (Pp 112; Sw Health Organization. 1 92-4-157141-1

Collagen remodelling in myocardia of patients with diabetes.

Abstract: AIMS: To investigate collagen remodelling in the interstitium of the heart in patients with diabetes. METHODS: Immunohistochemical study of the biopsied myocardium using type specific anticollagen antibodies (I, III, IV, V, VI) was performed in 12 patients with non-insulin dependent diabetes mellitus and six non-diabetic patients. There was no history of hypertension or coronary artery stenosis in any of the patients. RESULTS: Noticeable accumulations of collagen types I, III, and VI in the myocardial interstitium were recognised in both groups, but little accumulation of types IV or V was found. Types I and III mainly stained in the perimysium and perivascular region, while type VI predominantly stained in the endomysium. There was no disease specific accumulation of collagen in diabetes mellitus. The percentage of total interstitial fibrosis in the myocardium was significantly higher in the diabetic group than in the control group (p < 0.05). Although the percentages of collagen types I and VI did not differ between the two groups, the percentage type of III was significantly higher in the diabetic group than in the controls (p < 0.01). CONCLUSIONS: Collagen remodelling mainly as a result of an increase in collagen type III in the perimysium and perivascular region, occurs in the hearts of patients with diabetes.

Identification of Helicobacter pylori DNA in the mouths and stomachs of patients with gastritis using PCR.

Abstract: AIMS--To determine the prevalence of Helicobacter pylori colonisation in the mouths of patients with H pylori gastritis. METHODS--A nested polymerase chain reaction test for the 16S ribosomal RNA gene of H pylori was used on saliva, dental plaque, gastric juice and gastric biopsy specimens from patients attending a dyspepsia clinic. RESULTS--Thirteen patients had histologically confirmed Helicobacter associated gastritis. Twelve of these had positive gastric aspirates by PCR. Five had at least one positive oral specimen. Eight patients with normal gastric biopsy specimens had no PCR positive oral specimens or gastric aspirates. All, however, had PCR positive gastric biopsy specimens. In an attempt to determine the origin of these positive results in normal patients, it was shown that biopsy forceps could contaminate specimens with DNA from previous patients. CONCLUSION--The demonstration of the organism in the mouths of a substantial proportion of dyspeptic patients has major implications for the spread of H pylori and identifies a potential source for reinfection following eradication of the organism from the stomach.

Detection of Chlamydia pneumoniae and Chlamydia psittaci in sputum samples by PCR.

Abstract: AIMS--To use the polymerase chain reaction (PCR) to detect Chlamydia pneumoniae and Chlamydia psittaci in sputum samples. METHODS--A nested PCR was developed, the first stage of which amplified DNA from both C pneumoniae and C psittaci while the second stage targeted specifically at C pneumoniae, allowing the two species to be differentiated. The primers were designed not to amplify sequences from C trachomatis. A panel of 26 sputum samples from patients with community acquired pneumonia evaluated previously by enzyme linked immunosorbent assay (ELISA), direct immunofluorescence (DIF), and culture was tested blind by PCR. Most of these specimens also had accompanying serial serum samples which were tested for species specific antibodies using microimmunofluorescence (micro-IF). RESULTS--PCR detected C pneumoniae DNA in 10 of the 26 samples and C psittaci DNA in four. There was good concordance between ELISA, DIF, micro-IF and PCR in the C pneumoniae group. Two of the C psittaci identified by PCR were labelled C pneumoniae by DIF but the PCR results were supported by serology or a history of bird contact. Of the PCR negative group: six were true negative results; two contained C trachomatis. There were four discrepant results. CONCLUSIONS--The data suggest that PCR is effective in the detection of C pneumoniae. The sensitivity for C psittaci is inevitably lower due to the strategy taken but specificity seemed to be good.

Bile reflux and intestinal metaplasia in gastric mucosa.

Abstract: AIM: To determine associations between enterogastric bile reflux and gastric mucosal pathology. METHOD: A retrospective study using fasting gastric juice bile acid measurements and antral or prestomal biopsy specimens from 350 patients, 66 of whom had previously undergone surgery that either bypassed or disrupted the pyloric sphincter. RESULTS: Bile reflux was positively associated with reactive gastritis and negatively with Helicobacter pylori density. After stratification for previous surgery, age, and H pylori status, the histological feature most strongly associated with bile reflux was intestinal metaplasia, including all its subtypes. The prevalence of intestinal metaplasia was greatest in patients with both H pylori infection and high bile acid concentrations. Bile reflux was also positively associated with the severity of glandular atrophy, chronic inflammation, lamina propria oedema and foveolar hyperplasia. CONCLUSIONS: Bile reflux is a cause of reactive gastritis. It modifies the features of H pylori associated chronic gastritis. The changes are not confined to patients who have had surgery to their stomachs. The positive associations with atrophy and intestinal metaplasia have implications for models of gastric carcinogenesis.

Diagnosis of intestinal and disseminated microsporidial infections in patients with HIV by a new rapid fluorescence technique.

Abstract: AIMS--To assess the value of a new rapid fluorescence method for the diagnosis of microsporidiosis in HIV seropositive patients. METHODS--Microsporidian spores in stools were demonstrated by using the fluorochrome stain Uvitex 2B. The new technique was evaluated in three groups of HIV seropositive patients with diarrhoea. Group 1: 19 patients with biopsy confirmed E bieneusi infection (186 stool samples); group 2: 143 consecutive patients from whom faeces were submitted for routine investigation of diarrhoea (318 samples); group 3: 16 patients with small intestinal biopsy specimens negative for microsporidia (55 samples). The new method was used to monitor spore shedding during experimental treatment with paromomycin and albendazole in four patients. RESULTS--Brightly fluorescent spores were detected in all stool samples of patients in group 1. In group 2 16 (11%) patients had spores in their stool samples. E bieneusi was found in 11 patients; in the other five another genus of microsporidia, Encephalitozoon, was recognised. Encephalitozoon spores were also found in the urine of three of these patients and in the maxillary sinus aspirate of two of them, suggesting disseminated infection. The results were confirmed by electron microscopic examination. In group 3 negative biopsy specimens were confirmed by negative stool samples in all cases. Treatment with albendazole and paromomycin did not affect the spore shedding in three patients with E bieneusi infection. By contrast, in a patient with Encephalitozoon sp infection albendazole treatment resulted in clinical improvement together with complete cessation of spore excretion in the stool. CONCLUSION--The Uvitex 2B fluorescence method combines speed, sensitivity, and specificity for the diagnosis and treatment evaluation of intestinal and disseminated microsporidiosis.

Importance of apoptosis in the histopathology of drug related lesions in the large intestine.

Abstract: AIM: To investigate the possibility that the incidence of apoptotic bodies in the cryptal epithelium might help to identify colonic lesions due to drugs, especially non-steroidal anti-inflammatory drugs (NSAIDs). METHODS: The apoptotic count (AC) the number of apoptotic bodies per 100 crypts was calculated in a series of colorectal biopsy specimens, stained with haematoxylin and eosin from patients with (a) known or suspected drug induced colitis and (b) inflammatory bowel disease before or after treatment with salazopyrine or corticosteroids. These specimens were compared with normal biopsy specimens from a control group of comparable age and sex distribution. RESULTS: Under normal conditions apoptotic bodies were seldom seen at all and the mean apoptotic count was less than 1.0. In untreated inflammatory bowel disease the mean apoptotic count was marginally increased (2.4), but when there was a partial response to drug treatment the apoptotic count rose to 13.1 (p 0.003). In colonic lesions directly attributable to drugs the apoptotic count was always increased, reaching its highest level (106) with 5-fluorouracil. In colitis related to NSAIDs apoptoses were associated with inflammation, most notably an increase in lymphocytes in both lamina propria and epithelium. CONCLUSION: The presence of crypt apoptoses in substantial numbers (with an apoptotic count in excess of 5) should always raise the possibility of drug effect. The mechanisms involved are not clear but with NSAIDs the changes might well be immunologically mediated.

Method for grading breast cancer.

Abstract: labelling.' PCNA immunohistochemical expression (evaluated with the PCI0 monoclonal antibody) seems to be related to cellular proliferation in many normal tissues and in some neoplasms,2 such as gastrointestinal lymphomas,3 central nervous system tumours,4 lung neuroendocrine neoplasms,' and prostatic carcinomas.' However, in other tumours, like breast and gastric cancer, PCNA (PC 10) expression seems aberrant and not strictly related to proliferative activity.' 7 8 Various factors unrelated to cell proliferation may influence the immunohistochemical expression of PCNA, including posttranscriptional regulation (and deregulation) of the PCNA gene,8\" long half-life of the PCNA protein,'0 involvement of PCNA protein in DNA repair synthesis,\" and tissue and section processing-type and ionic strength of the fixatives, fixation time, section heating, immunohistochemical techniqus8 . 2 1 3 ques. Further problems in PCNA immunohistochemical staining, as in other kinetic quantitative immunohistochemical studies, concern evaluation and scoring methods. '4 15 Should we use quantitative or semiquantitative methods? How many cells should be counted? Which tumour areas should be evaluated (the most positive or random selected areas)? Which immunoreactive cells should be evaluated (all positive cells or only the most intensely stained)? Particular attention should be also drawn to the kind of antibody used to localise PCNA. Different staining patterns may be seen with different antibodies, and this may add to conflicting and confusing results.'4 In our opinion PCNA immunostaining should be evaluated with great caution and in some fields even with scepticism. More work is needed to assess the extent and range of PCNA staining in different tissues and lesions (neoplastic and non-neoplastic). PCNA counts should be evaluated concurrently with the different anti-PCNA available antibodies and the results should be compared with other \"proliferation markers\" and especially with clinical data. The possibility that PCNA immunostaining may have diagnostic5 or prognostic value7 is intriguing and carefully performed clinicopathological studies are needed to assess this possibility further. This will be the only way to know if we are faced with an interesting but clinically worthless tool or with an important test to be added to the routine evaluation of neoplasms.

Neoplasms with Follicular Differentiation

Abstract: If you wish to order, or require further information regarding the titles reviewed here, please write or telephone the BMJ Bookshop, PO Box 295, London WC1H 9TE. Tel: 071 383 6244. Fax: 071 383 6662. Books are supplied post free in the UK and for British Forces Posted Overseas addresses. Overseas customers should add 15% for postage and packing. Payment can be made by cheque in sterling drawn on a UK bank, or by credit card (MasterCard, VISA, or American Express) stating card number, expiry date, and your full name.

Increased pentane and carbon disulfide in the breath of patients with schizophrenia.

Abstract: AIMS--To determine the concentrations of pentane (a marker of lipid peroxidation) and other volatile organic compounds in the breath of patients with schizophrenia. METHODS--Volatile organic compounds were assayed by gas chromatography/mass spectroscopy (GC/MS) in 88 subjects--25 with acute schizophrenic psychosis, 26 with psychiatric disorders other than schizophrenia, and 37 normal volunteers. RESULTS--The mean alveolar gradients of pentane and carbon disulfide (CS2) were significantly higher in the patients with schizophrenia than in the control groups. CONCLUSIONS--Schizophrenia may be accompanied by accelerated lipid peroxidation in cell membranes, as well as increased manufacture of CS2, a known neurotoxin.

Expression of gelatinase A and TIMP-2 mRNAs in desmoplastic fibroblasts in both mammary carcinomas and basal cell carcinomas of the skin.

Abstract: AIMS--To compare the localisation of mRNAs for the basement membrane degrading enzyme gelatinase A (72 kilodalton type IV collagenase) and its inhibitor TIMP-2 in carcinomas of the breast and basal cell carcinomas of the skin which have little or no ability to metastasize. METHODS--In situ hybridisation was performed on formalin fixed, paraffin wax embedded blocks using 35S-labelled riboprobes on 16 mammary carcinomas, three fibroadenomas, and a benign phyllodes tumour, and on 15 basal cell carcinomas of the skin (BCC). RESULTS--Labelling for both mRNAs was detectable in 14 of 16 mammary carcinomas and in 13 of 15 BCC, most often over organising desmoplastic fibroblasts in the stroma around invasive epithelial aggregates. Some sparse labelling was seen over malignant epithelial cells in six of the mammary carcinomas but not in the BCC. Some expression of gelatinase A mRNA was also seen in fibroblasts of breast lobules adjacent to the mammary carcinomas and around engulfed adnexal elements in the BCC, but not in unaffected breast tissues, fibroadenomas, the phyllodes tumour or unaffected skin. CONCLUSIONS--Maximal expression of gelatinase A and TIMP-2 mRNAs occurs in malignant neoplasms as part of the host response to the presence of established neoplastic cells rather than as an initial response to invasion. The degree to which this is present suggests this may be a highly relevant mechanism modulating tumour differentiation, growth and progression, possibly entailing uptake via specific receptors on the tumour cell surface.

Cytokines in stools of children with inflammatory bowel disease or infective diarrhoea.

Abstract: AIMS--To determine the concentrations of interleukin-6 (IL-6) and tumour necrosis factor alpha (TNF alpha) in stools from children. METHODS--Stool samples from 14 healthy children, 32 children with inflammatory bowel disease, and 23 children with acute diarrhoea were emulsified in an equal volume of phosphate buffered saline and then centrifuged to produce a clear supernatant fluid. IL-6 and TNF alpha were measured by enzyme linked immunosorbent assay (ELISA). RESULTS--TNF alpha was detected in the stools of all 14 healthy children (12-130 pg/g stool), but IL-6 was detected only in three. Similar results were seen in children with inactive inflammatory bowel disease. Stool TNF alpha concentrations were raised in samples from children with active inflammatory bowel disease, but in most (11/18) of these samples IL-6 was undetectable. Stool samples contained a heat-labile factor which rapidly destroyed IL-6 immunoreactivity. Most children with diarrhoea had TNF alpha concentrations similar to those of healthy controls and most were also negative for IL-6. Three children with Shigella flexneri infection had extraordinarily high concentrations of both TNF alpha and IL-6 in their stools. CONCLUSIONS--There is constant low grade production of TNF alpha in the intestine of healthy people. Raised values are indicative of mucosal inflammation, but are not specific. Stool IL-6 is of little use in assessing mucosal inflammation because immunoreactivity is rapidly lost in stool samples.

The Correspondence of Charles Darwin

Abstract: From Francis Darwin [1875?]1 Down My dear Father I have had two mornings work at Drosera but without success. I got into a very good way of doing it but the plants seem sluggish. The first morning 26′′ was the quickest—counting from the beginning of contact of the drop of meat infusion. The second morning they were more sluggish still; ammonia is not so nice to work with as meat; as with meat one is not afraid of evaporation making ones drop weak if one waits a bit—2 The only thing of the slightest interest is that contact for 1′ produced movement in just the same time counting from the beginning as contact for 4′′ did; the tentacles were on the same leaf; but of course one experiment isnt much good— I shall try again, because now I can do it accurately— My frog preparations are pretty good—3 I am very glad mother & you are keeping well.4 Thank you for writing about the pamphlets, we have cut up a lot & sorted some— it will be done when you come home— Yr affec son | F Darwin

Abnormal growth factor and cytokine expression in Dupuytren's contracture.

Abstract: AIM--To analyse patterns of gene expression for peptide regulatory factors in patients with Dupuytren's contracture. METHODS--Tissue samples (palmer fascia) from 12 patients with Dupuytren's contracture and 12 controls were studied using the reverse transcription/polymerase chain reaction (RT/PCR) technique. RESULTS--Tissue from patients with Dupuytren's contracture expressed a higher percentage of peptide regulatory factors than that of controls: interleukin-1 alpha (83% v 16%; p < 0.01); interleukin-1 beta (66% v 8%; p < 0.01); transforming growth factor beta (75% v 25%; p < 0.02); and basic fibroblast growth factor (66% v 25%; p < 0.05). Platelet derived growth factors alpha and beta were also expressed more commonly (66% v 33% and 25% v 16%, respectively), but these differences were not significant. CONCLUSIONS--The increased prevalence of expression for the above mRNAs in Dupuytren's tissue is relevant as interleukin-1, basic fibroblast growth factor, and transforming growth factor beta stimulate the growth of fibroblasts and transforming growth factor beta also enhances production of collagen and other extracellular matrix proteins. Excessive local release of these peptide regulatory factors may have an important role in the pathogenesis of Dupuytren's contracture.

Plasma coenzyme Q (ubiquinone) concentrations in patients treated with simvastatin.

Abstract: Plasma coenzyme Q (CoQ) was measured in 20 hyperlipidaemic patients treated with diet and simvastatin (an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase); 22 hyperlipidaemic patients treated with diet with alone; and 20 normal controls. Patients treated with simvastatin had a significantly lower plasma CoQ and CoQ: cholesterol ratio than either patients receiving diet alone or normal controls. Use of simvastatin was inversely and independently correlated with both CoQ (p < 0.0001) and CoQ: cholesterol ratio (p < 0.01). There was a significant inverse association between CoQ and dose of simvastatin (p < 0.001). It is concluded that simvastatin may lower the plasma CoQ concentration and this may be greater than the reduction in cholesterol. The possible adverse effect of simvastatin on the metabolism of CoQ may be clinically important and requires further study.

Prospective comparative study of computer programs used for management of warfarin.

Abstract: AIMS--To compare the effectiveness of three computerised systems that are currently used for assisting warfarin control in outpatients with the customary dosing method used by experienced medical staff. METHODS--A pilot randomised study of three systems with a follow up independently randomised study of two of these was made on 186 patients receiving long term treatment or who had recently started warfarin treatment and had been discharged from hospital. RESULTS--All three computerised systems seemed to give satisfactory control compared with the traditional dosing method. For patients receiving more intensive treatment with an assigned target range of 3.0-4.5 computerised dosage programs achieved significantly better control; the medical staff undertreated such patients almost 50% of the time. CONCLUSION--Computer based programs can assist outpatient anticoagulant control with warfarin during both early and long term treatment. For most patients the control achieved is as good as that obtained by the customary method of dosing, by experienced clinic doctors, although the latter tend to be too conservative when dosing patients within the intense target range of 3.0 to 4.5 International Normalised Ratio (INR). The computers were significantly more successful in this higher range.

Out of Africa: observations on the histopathology of Mycobacterium ulcerans infection.

Abstract: Introduction Mycobacterium ulcerans infection produces progressive skin ulceration in man and other mammals. The disease was first described in 1948 in patients from the Bairnsdale district in Australia' but was known in Africa well before this time.2 It has since been recorded in many, mostly tropical countries in Africa, Central and South America, and South East Asia.3 The infection characteristically occurs in defined areas which are associated with river or lacustrine systems draining tropical or warm-temperature rain forest. Like the flora to which it is related, the mycobacterium shows a Gondwanian distribution which is evidence of its great antiquity.4 The disease usually begins as a skin papule (stage IA) which later ulcerates (stage 2) (fig 1)5; less commonly the infection may first appear as a subcutaneous nodule (stage 1B)6 which subsequently affects the dermis and then ulcerates, or sometimes the disease may produce diffuse oedema of a limb (stage 1 C),7 due to a necrotising panniculitis, which clinically mimicks cellulitis. The infection is usually on an extremity but, particularly in children, it may affect the face or trunk.8 Occasionally, the necrotic process extends through deep fascia with involvement of muscle9 and even bone.'0 Metastatic infection may occur. The features of the pathology ofM ulcerans infection have been described'; additional characteristics of the disease, as observed in Zaire (Belgian Congo), have been detailed in the papers by Janssens et al.7" l Infection in Uganda has been described by Dodge,'2 13 and later by Connor and Lunn,'4 15 and infection in Papua New Guinea by Lytton and Lavett. 6 Hayman and McQueen described the pathology in additional Bairnsdale district patients in 1985.1' Burchard and Bierther'8 briefly described the pathology of the disease as seen in Gabon; their paper was illustrated with electron photomicrographs of M ulcerans. The earlier descriptions of the pathology of the disease have been comprehensively reviewed. '9 In the original paper' MacCallum detailed the clinical and macroscopic appearances of the ulcers with straight or undermined edges, subcutaneous spread producing nodules of induration and eventual ulceration at a distance from the initial lesion (fig 1), and the development in the subcutaneous tissue of "an abundant gelatinous mass like blubbery granulation tissue which could be readily wiped off with gauze". Histologically, necrotic tissue lined the ulcer wall and floor and extensive necrosis of fatty tissue extended beneath the Figure 1 Clinical appearances of Mycobacterium ulcerans infection, on the posterolateral aspect of the left leg in a 64 year old man. The photograph demonstrates the undermining of skin and the development of a satellite lesion associated with extensive necrosis of subcutaneous fat. (Photograph, courtesy ofMr Kendall Francis, FRACS.).

Can histopathologists reliably diagnose molar pregnancy

Abstract: AIMS--To assess the degree of difficulty in diagnosing partial mole by analysing intraobserver and interobserver agreement among a group of pathologists for these diagnoses. METHODS--Fifty mixed cases of partial mole, complete mole, and non-molar pregnancy were submitted to seven histopathologists, two of whom are expert gynaecological pathologists; the other five were district general hospital consultants, one of whom works in Australia. These participants gave each slide a firm diagnosis of either partial mole, complete mole, or non-molar pregnancy. Some 12 months later, the slides were recorded and again submitted for a second diagnostic round to assess intraobserver as well as interobserver agreement. Standard histological criteria for each diagnostic category were circulated with the slides. RESULTS--kappa statistics showed that complete mole could be reliably distinguished from non-molar pregnancy, but neither non-molar pregnancy nor complete mole could be easily differentiated from partial mole. In only 35 out of 50 cases was there agreement between five or more of the seven participants. Agreement between the expert gynaecological pathologists was no better than for others in the group. Interestingly, the intraobserver agreement for each pathologist was good to excellent. CONCLUSIONS--These results imply that the reported histological criteria are either not being applied consistently or that they are lacking in practical use. An atypical growth pattern of trophoblast, rather than the polar accentuation seen in normal first trimester pregnancies, seems to be the important diagnostic histological feature for partial mole. Ploidy studies might also help with problem cases.

Prevalence of lymphoid follicles and aggregates in Helicobacter pylori gastritis in antral and body mucosa.

Abstract: AIMS--To evaluate the prevalence of lymphoid follicles and aggregates in the antral and body mucosa in Helicobacter pylori gastritis and to assess if there were correlations with ulcers in the duodenum, pylorus, or stomach, and with chronic antral erosions. METHODS--Patients (n = 2692) with histologically confirmed H pylori antral gastritis were investigated. These comprised five groups: those with duodenal ulcers; those with pyloric ulcers; those with gastric ulcers; those with chronic erosions; and those with no associated lesions. In 1446 cases at least two additional biopsy specimens from the oxyntic mucosa were available. RESULTS--Lymphoid follicles and aggregates were found in 53.8% of cases in the antral mucosa compared with 14.8% in the oxyntic mucosa (p < 0.001). The various diseases showed significant differences in terms of the prevalence of follicles and aggregates: The highest numbers in the antral mucosa as well as the lowest in the oxyntic mucosa were found in patients with duodenal ulcers (60.5% and 9.2%, respectively). The highest numbers of follicles and aggregates in the oxyntic mucosa occurred in patients with gastric ulcers. CONCLUSIONS--The detection of lymphoid follicles and aggregates in oxyntic mucosa and the higher prevalence in antral mucosa fits well with the distribution of primary gastric lymphomas. This adds further weight to the notion that the development of follicles and aggregates, triggered by H pylori, might be an early precursor to gastric lymphoma. The differences between the groups investigated might be due to different strains of H pylori or differences in the respective sizes of antral and oxyntic mucosa.

Prognostic value of epidermal growth factor receptor expression in cervical carcinoma.

Abstract: AIMS: To investigate the pattern of epidermal growth factor receptor expression and its prognostic value in the three main types of cervical carcinoma. METHODS: 62 cases of stage IB/IIA cervical carcinoma, all with a minimum of five years of follow up, were studied. Representative sections were stained for mucin to permit accurate tumour typing and a standard avidin-biotin immunoperoxidase technique using the polyclonal antibody 12E was used to demonstrate the presence of epidermal growth factor receptor. RESULTS: A proportion of all three tumour types expressed epidermal growth factor receptor, it being most common in squamous cell carcinomas (50%). Overall, there was a correlation between epidermal growth factor expression and mortality. This was particularly obvious in the absence of lymph node metastases. When the individual tumour types were considered this association with prognosis was not demonstrable for squamous cell carcinomas or adenocarcinomas but was a very prominent feature of adenosquamous carcinomas. CONCLUSIONS: Immunohistochemical demonstration of epidermal growth factor receptor expression may be useful in identifying those patients with a poor prognosis, particularly those with adenosquamous carcinomas which have not metastasised to the regional lymph nodes.

ACP Broadsheet no 137: April 1993. Obtaining samples at post mortem examination for toxicological and biochemical analyses.

Abstract: Introduction The human body can be regarded as a complex assembly of dynamic chemical systems. Maintaining the integrity of these systems is an affront to the second law of thermodynamics and requires the constant expenditure of energy. Once death takes place, the supply of energy from metabolic processes is dramatically reduced, the integrity of the different compartments within the body breaks down at differing rates; complex molecules tend to break down to their simpler subunits and to move down concentration gradients that were maintained in life by the expenditure of metabolic energy. Obviously, these processes do not all occur at once. Thus, for a variable length of time after death the analysis of appropriate samples may yield useful information about the metabolic state of the person in the period immediately before death. Once death has taken place many drugs are released from their binding sites in tissue as pH decreases on death and as the processes of autolysis proceed. These phenomena can make the interpretation of drug concentrations after death less than straightforward.

New antiserum against Ki-67 antigen suitable for double immunostaining of paraffin wax sections.

Abstract: AIMS--To prepare a rabbit antiserum equivalent to MIB 1 to permit the simultaneous assessment of cell proliferation and other markers of interest using double labelling studies. METHODS--Rabbits were immunised with a synthetic peptide deduced from the cDNA sequence coding for the Ki-67 antigen. Serum samples were tested for immunoreactivity using different immunobiochemical methods. RESULTS--A polyclonal antiserum was derived which detects the native as well as recombinant parts of the Ki-67 antigen in different test systems. Furthermore, the antiserum stains the Ki-67 antigen in routinely processed, paraffin wax embedded material. CONCLUSIONS--After antigen unmasking by microwave treatment the antiserum described here represents a powerful tool for the determination of growth fractions even in archival material. It is especially suitable for double staining experiments in combination with monoclonal antibodies.

Evaluation of high performance liquid chromatography for routine estimation of haemoglobins A2 and F.

Abstract: AIMS--To compare high performance liquid chromatography (HPLC) with conventional methods for the estimation of blood haemoglobin A2 (HbA2) and haemoglobin F (HbF) concentrations in routine thalassemia screening. METHODS--An HPLC system (the VARIANT Hemoglobin Testing System) was tested for precision and reproducibility in the measurement of HbA2 and HbF, and reference ranges were obtained for a local healthy adult population. HPLC was compared with column anion exchange chromatography for HbA2 measurement, and radial immunodiffusion, or alkaline denaturation for HbF measurement. The reliability of HbA2 measurement by HPLC for the detection of beta thalassaemia and HbE was assessed in 200 consecutive samples for routine thalassaemia screening. RESULTS--HPLC was rapid, technically easy, and gave good precision and reproducibility. In all comparisons linear regression analysis showed good correlation between HbA2 or HbF concentrations determined by HPLC and by the respective conventional methods. All beta thalassaemia or haemoglobin E carriers presumptively identified by conventional methods in 200 consecutive samples were detected by HbA2 measurement using HPLC, without any false positive or false negative results. CONCLUSIONS--The measurement of HbA2 and HbF by HPLC is rapid, reproducible, and precise. It is as reliable as column chromatography for the measurement of HbA2 and radial immunodiffusion or alkaline denaturation for the measurement of HbF. HPLC may be an appropriate method for rapid screening in population surveys for beta thalassaemia and HbE carriers.

Hematology of Infancy and Childhood

Abstract: The hematology of infancy and childhood that we provide for you will be ultimate to give preference. This reading book is your chosen book to accompany you when in your free time, in your lonely. This kind of book can help you to heal the lonely and get or add the inspirations to be more inoperative. Yeah, book as the widow of the world can be very inspiring manners. As here, this book is also created by an inspiring author that can make influences of you to do more.

The British Society for Haematology Guidelines on the use and monitoring of heparin 1992: second revision. BCSH Haemostasis and Thrombosis Task Force.

Abstract: CHEMISTRY OF HEPARIN Unfractionated heparin is a naturally occurring glycosaminoglycan produced by the mast cells of most species. It is extracted from porcine or bovine mucosa, all the products currently used in the United Kingdom being of porcine origin. It consists of alternating chains of uronic acid and glucosamine, sulphated to varying degrees, and has a molecular weight range of 5000-35 000, with a mean of about 12 000-14 000. Low molecular weight heparin is manufactured from unfractionated heparin by controlled depolymerisation using chemical (nitrous acid or alkaline hydrolysis) or enzymatic (heparinase) methods. Although these processes yield different end groups, there is no evidence that these differences in chemical structure affect biological function. The biological properties of any low molecular weight heparin are primarily determined by its molecular weight distribution. The products currently available have an average molecular weight between 4 000 and 6 000, and 60-80% of the total polysaccharides lie between 2 000 and 8 000. greater than the anti-IIa or APTT activity. In the low molecular weight heparins currently available the ratio of anti-Xa to anti-Ila activity ranges from 1-8 to 3*5.

Immunoglobulin and complement deposition in glomeruli of 756 subjects who had committed suicide or met with a violent death

Abstract: AIMS--To study immune deposits in renal glomeruli. METHODS--Tissue was obtained from 756 necropsy cases from people who had committed suicide or met with a violent death. Glomerular immune deposits were examined by immunofluorescence microscopy and a light microscopy. The clinical histories of all the decreased were studied to ascertain reasons for the deposits. RESULTS--Immune deposits were found in glomeruli in 91 (12%) cases. In 52 (6.8%) cases mesangial IgA was observed as a solitary finding in 34 (4.5%), and was accompanied by other immunoglobulins in 18 (2.4%). Mesangial IgM was present in 19 (2.5%) and IgG in 11 cases (1.5%). Two cases had capillary IgG (0.3%). Light microscopic examination showed mesangial enlargement in eight of the cases with mesangial IgA. These included one with IgA glomerulonephritis diagnosed before death. Two cases with normal glomerular morphology and mesangial IgA deposits had clinical laboratory evidence of renal disease. In two subjects with normal glomerular morphology, mesangial IgM and microscopic haematuria were present. In one case with capillary IgG membranous glomerulonephritis was detected. CONCLUSIONS--Ten cases had mesangial IgA together with morphological or clinical laboratory findings suggestive of renal disease. If all these are regarded as IgA glomerulonephritis, then its prevalence can be estimated at 1.3%. For IgM glomerulonephritis, a prevalence of 0.3% was deduced.

Overexpression of p53 protein in Barrett's syndrome with malignant transformation.

Abstract: AIMS--To study the overexpression of p53 protein in Barrett's oesophagus with adenocarcinoma, and to correlate this expression with the pathological features of Barrett's syndrome. METHODS--Immunohistochemical staining was performed on frozen sections with a monoclonal antibody directed against wild type and mutated p53 protein (Pab 1801). Eleven cases of Barrett's adenocarcinoma were studied, seven of which had extensive sampling of benign Barrett's mucosa. RESULTS--Eight of 11 adenocarcinomas overexpressed the p53 protein. Both early and advanced tumours were positive. In Barrett's mucosa around the p53 positive tumours, high grade dysplasia was positive; low grade dysplasia and non-dysplastic mucosa were negative. CONCLUSIONS--P53 gene mutation with ensuing p53 protein overexpression is a common feature of Barrett's adenocarcinoma, both at early and advanced stages. This mutation appears as a relatively late event during the neoplastic transformation of Barrett's oesophagus.

Negative cytology preceding cervical cancer: causes and prevention.

Abstract: AIM--To assess the validity of negative cervical smear reports in women who subsequently developed cervical cancer; and to determine means of improving the screening process. METHODS--One hundred and forty cervical smears, initially reported as negative from 103 women, and taken up to 12 years before diagnosis of cervical cancer, were reviewed. RESULTS--Ninety two smears contained dyskaryotic cells. Analysis showed that these smears formed several well defined patterns. False negative reports were likely to occur if fragments of neoplastic tissue rather than dissociated dyskaryotic cells were present or if the smear contained few dyskaryotic cells. Screening fatigue appeared to be a factor in others. It was also considered important that smears contained cells from the endocervix. These were deficient in 64% of the 47 smears confirmed as negative on review and in 69% of smears containing only a few dyskaryotic cells. CONCLUSIONS--Current methods of quality assurance will not remedy these defects in the screening process. It is the responsibility of laboratories to identify sources of poor smears and liaise with smear takers to ensure an improvement in quality. Assessment of the quality of smears received by a laboratory should become an important part of audit. Staff training should place more emphasis on the interpretation of "microbiopsies". The adoption of a quick scanning technique before conventional screening would probably also substantially reduce false negative results.

Expression of c-myc and bcl-2 oncogene products in Reed-Sternberg cells independent of presence of Epstein-Barr virus.

Abstract: AIMS: To evaluate the expression of c-myc and bcl-2 oncogene products in Reed-Sternberg cells in Hodgkin's disease, especially in relation to Epstein-Barr virus infection and expression of EBV encoded latent membrane protein (LMP). METHODS: Tissues from 33 cases of Hodgkin's disease were studied for the presence of EBV DNA by polymerase chain reaction (PCR) and DNA in situ hybridisation (DISH), for the presence of EBER-1 and EBER-2 EBV RNA by RNA in situ hybridisation (RISH); and for the presence of LMP, bcl-2, and c-myc proteins by immunohistochemical staining. RESULTS: A substantial number of Reed-Sternberg cells expressed bcl-2 in 20 of 29 (69%) and c-myc in 30 of 32 (94%) Hodgkin's disease samples. In 18 of the 25 (72%) cases Reed-Sternberg cells expressed both oncogene products. Of these 18 cases, 10 (56%) were EBV-PCR positive; eight (44%) were EBV-PCR negative. CONCLUSIONS: Reed-Sternberg cells in Hodgkin's disease frequently express both bcl-2 and c-myc oncogene products, suggesting that these oncogenes may act in concert in the pathogenesis of the disease. Moreover, the expression of c-myc and bcl-2 proteins in Reed-Sternberg cells is independent of EBV and LMP status.