bluuebunny/crossref_metadata_2025 huggingface dataset:
Atheroembolism.
Authors: Kealy W F
Published: June 2007

A review of the literature has shown that atheroembolism as a cause of clinically evident disease is an entity that is little documented. Sections of tissues from necropsies on patients over the age of 60 years from 1970 to 1977 inclusive were reviewed and examined for evidence of atheroembolism, and an incidence of 0.79% was found. In every case of embolism the aorta showed advanced atheroma, sometimes with aneurysm formation. The ESR in some instances was increased, sometimes markedly so. Some of the problems of antemorten diagnosis are discussed. To emphasise the possible clinical importance of the condition, a rare instance of spinal cord infarction due to atheroembolism is described.
Sensitivity of immunoperoxidase and immunofluorescence staining for detecting chlamydia in conjunctival scrapings and in cell culture.
Authors: Woodland R M , El-Sheikh H , Darougar S , Squires S
Published: June 2007

The sensitivities of Giemsa, immunofluorescence, and immunoperoxidase staining for the detection of Chlamydia psittaci inclusions in conjunctival scrapings and in irradiated McCoy cell monolayers were compared. Conjunctival specimens were obtained from a cat colony in which a trachoma-like disease, feline chlamydial keratoconjunctivitis, was endemic. The two immunochemical techniques were found to be of equal sensitivity and 50% to 100% more sensitive than Giemsa stain. Permanent preparations of immunoperoxidase stained material can be made and can be read using a simple light microscope. These features make the technique more useful than immunofluorescence staining, which gives temporary preparations that must be examined with a specialised fluorescence microscope.
Hepatitis B virus antigens in primary hepatic carcinoma: immunofluorescent techniques on fixed liver tissue.
Authors: Trevisan A , Realdi G , Losi C , Ninfo V , Rugge M , Rampinelli L
Published: June 2007

The presence of hepatitis B surface (HBsAg) and core (HBcAg) antigens was investigated by immunofluorescence in specimens of liver tissue obtained at necrospy in 107 patients with primary hepatic carcinoma. HBsAg was detected in the cytoplasm of liver cells in 16 cases, and in eight of them the antigen was also found in malignant cells. HBcAg, which was present in the nuclei of liver cells in eight cases, was detected in the nuclei of tumour cells in six of these and also in two other cases showing HBsAg, but not HBcAg, in the nonneoplastic tissue. Although most of the primary hepatic carcinomas studied were associated with cirrhotic changes in the non-neoplastic tissue, HBsAg and HBcAg were also detected in the absence of underlying cirrhosis. Hepatitis B virus markers were demonstrated in non-neoplastic tissue, mainly in patients with a well-differentiated carcinoma, and only in these cases were they found also in the neoplastic tissue. These results show that hepatitis B virus antigens, including HBcAg, can be detected in the neoplastic cells of well-differentiated carcinoma of the liver. Although these cells could have been infected after the malignant transformation, a direct oncogenic role of the virus cannot be excluded.
Evaluation of dithiothreitol (DTT) for inactivation of IgM antibodies.
Authors: Okuno T , Kondelis N
Published: June 2007

A newly introduced sulphydryl compound, dithiothreitol (DTT), is evaluated for its optimal conditions of inactivation of IgM antibodies. The maximal effects of DTT reagent are observed when its final concentrations are between 0.0025 M and 0.005 M, pH between 7.0 and 8.0, and incubation at 37 degrees C. Concentrations over 0.01 M, pH values greater than 8.0, and an incubation temperature over 40 degrees C resulted in a gel formation of the specimen. Examination of both 'cold' and 'warm' type antibodies demonstrated that the results obtained by the DTT reagent are in close agreement with those obtained by 2-mercaptoethanol reagents and DEAE Sephadex treatment. Since the procedure is simple and rapid and lacks offensive odour, DTT is recommended for routine use in blood banking for the inactivation of IgM antibodies.
Latex agglutination, counterimmunoelectrophoresis, and protein A co-agglutination in diagnosis of bacterial meningitis.
Authors: Dirks-Go S I , Zanen H C
Published: June 2007

Specimens of cerebrospinal fluid from 201 patients with meningitis caused by Neisseria meningitidis groups A/B/C/135, Streptococcus pneumoniae (23 types), and Haemophilus influenzae type b were tested for the presence of specific bacterial antigens by latex agglutination, counterimmunoelectrophoresis (CIE), and protein A co-agglutination. Specific antigens were found in 75% of the specimens by latex agglutination and CIE, and in 60% of the specimens by protein A co-agglutination. Non-specific reactions in protein A co-agglutination were prevented by heating the specimens to 100 degrees C for a few minutes. The three methods are simple and quick to perform. The smallest amount of antiserum was used in protein A co-agglutination, but we found this method less sensitive. Latex agglutination and CIE proved to be equally sensitive, but the first method was easier to adopt in practice.
Meningococcal infection and proteolytic control.
Authors: Stark J M , Matthews N , Ryley H C , Greenwood B M , Lewis L S , Whittle H C
Published: June 2007

Cascade enzyme inhibitors (C1-esterase inhibitor, C3b inactivator, antithrombin III) and other major proteolytic enzyme inhibitors (alpha 1 trypsin inhibitor, alpha 1 chymotrypsin inhibitor, inter-alpha-trypsin inhibitor, alpha 2 macroglobulin) as well as C3 and alpha 1 acid glycoprotein, have been examined in the sera of Nigerian patients suffering from meningococcal infection of varied severity. Patients with meningococcaemia had lower serum concentrations of important inhibitors than did patients with localised meningitic infection. Within the coccaemic group, those who died had the lowest values, notably of antithrombin III and alpha 2 macroglobulin (and also of C3). The clinical end-result of meningococcal infection may be related to the degree of disequilibrium of the linked system of proteolytic control induced by the meningococcal endotoxin.
Carriage of group D streptococci in the human bowel.
Authors: Noble C J
Published: June 2007

Sixty faecal samples, 39 from adults and 21 from neonates, were investigated by means of a simple scheme to isolate and identify any group D streptococci present. A number of differences were found between the two groups. Group D streptococci were absent from 43% of the neonates compared with only 13% of the adults; Streptococcus bovis was commoner in the neonates (23.8%) than in the adults (5%), and Streptococcus faecium was not isolated from any of the infant samples although it was common in the adult samples (25%). The viable counts of Strep. faecium were found to be, on average, 100-fold lower than those of Streptococcus faecalis. The methods and results are discussed with reference to the clinical significance of group D streptococci, especially in bacterial endocarditis and carcinoma of the colon.
Effect of calcitonin treatment on osteoclast counts in Paget's disease of bone.
Authors: Williams C P , Meachim G , Taylor W H
Published: June 2007

Histological and biochemical changes during calcitonin treatment have been studied in 15 patients with Paget's disease of bone. For each patient, osteoclast counts were made by the same observer on serial needle biopsies of diseased bone from the posterosuperior iliac spine. Serial estimations were also made of the serum alkaline phosphatase and urinary hydroxyproline excretion. A total of 66 biopsies was examined (ranging from two to seven per patient). Osteoclast populations and the biochemical measurements were log normally distributed. During calcitonin treatment there was a statistically significant decrease in: (1) the total osteoclast count per square millimetre; (2) the number per square millimetre of osteoclasts in resorption cavities on the trabecular surface; (3) the relative proportion of osteoclasts sited in resorption cavities compared with total osteoclasts; (4) the serum alkaline phosphatase level; (5) 24-hour urinary hydroxyproline excretion. On stopping treatment there was a statistically significant increase in all of these histological and biochemical values except that the proportion of osteoclasts in resorption cavities remained low. The trabecular cement line pattern remained abnormal during and after treatment in all biopsies examined, and complete suppression of osteoclast activity was not observed. One of the patients developed a Paget's osteosarcoma while on calcitonin therapy.
Congenital dyserythropoietic anaemia with unusual cytoplasmic inclusions.
Authors: Kenny M W , Ibbotson R M , Hand M J , Tector M J
Published: June 2007

A case of congenital dyserythropoietic anaemia is described which appears to be at variance with the accepted classification. The morphological findings are of normoblastic hyperplasia with marked cytoplasmic vacuolation. Electron microscopy shows these to be myelin figures in all stages of erythroid differentiation.
Comparative study of the endotoxin-stimulated nitroblue tetrazolium test in disease and health.
Authors: Haim N , Obedeanu N , Meshulam T , Robinson E , Ben-Layish E , Merzbach D
Published: June 2007

Endotoxin-stimulated nitroblue tetrazolium (NBT) reduction was evaluated in 853 individuals: 270 healthy controls, 334 with various non-neoplastic conditions, 220 with solid non-lymphomatous tumours, and 29 with lymphoma. Each of the above groups was divided into three age subgroups: less than 60, 60-69, and greater than or equal to 70 years. In the controls and in patients with nonmalignant diseases, significantly lower values were recorded for elderly subjects (greater than or equal to 60 years) compared with younger subjects of the same group, whereas in cancer patients the results were independent of age. Under the age of 60, stimulated values for both patient groups were significantly lower than the control values, and in patients with solid non-lymphomatous tumours significantly lower values were attained than in the other patients. NBT reduction in lymphoma patients was comparable to that of the controls. In the elderly (age greater than or equal to 70) no significant differences were noted between patients and controls. It is suggested that stimulated NBT reduction declines with advancing age. While this test clearly demonstrates some leucocyte dysfunction in solid cancer, its value in investigating neutrophil behaviour in elderly subjects is questioned.
Crossreaction of antilymphocyte globulin with human granulocyte colony-forming cells.
Authors: Barrett A J , Longhurst P , Rosengurt N , Hobbs J R , Humble J G
Published: June 2007

Clinical preparations of horse antilymphocyteglobulin (ALG) were found to inhibit human bone marrow granulocyte colony growth. This effect was enhanced by complement and was dose dependent, being almost complete at ALG concentrations of 100 microgram/ml. Inhibition was a property of ALG but not of normal horse globulin. However, short incubation of ALG with bone marrow cells occasionally stimulated colony growth and normal horse globulin regularly stimulated it. Three hours' incubation of bone marrow cells with ALG was needed to produce consistent colony inhibition, which was measurable as a reduction in the expected number of colonies and as a fall in the colony: cluster ratio of surviving cell aggregates. Absorption of ALG on acute myeloid leukaemia blast cells removed the inhibiting property of the ALG while preserving its lymphocytotoxic action. Serum from two patients receiving ALG treatment inhibited colony growth for up to 48 hours after ALG administration. The results suggest the presence in ALG of antibodies specifically cytotoxic to myeloid stem cells which may relate to its myleosuppressive properties in vivo, and also indicate that it should be possible to remove antimyeloid antibodies from ALG by absorption. The use of such purified ALG would have advantages in clinical bone marrow transplantation.
The case for laboratory aides in district clinical chemistry departments.
Authors: Watson D
Published: June 2007

A case of need is made out for the general introduction into district clinical chemistry departments of the existing experimental grade of laboratory aide or assistant. Subject to direct supervision by qualified personnel the aide has a useful role to fulfil. Important advantages to staff and patients accrue from the employment of this semiskilled laboratory worker.
Sporicidal activity of mixtures of alcohol and hypochlorite.
Authors: Coates D , Death J E
Published: June 2007

In an investigation of the activity against Bacillus subtilis spores of various mixtures of alcohol and sodium hypochlorite 50% methanol with enough hypochlorite to provide 2000 parts per million available chlorine water was the most sporicidal. This mixture, both when freshly prepared and for at least eight hours after mixing, achieved a five-log reduction in spore count in 10 minutes. The sporicidal activity of mixtures containing the same initial level of available chlorine was found to vary with the alcohol, initial alcohol concentration, and age of the mixture. Such mixtures may be useful for disinfecting heat-sensitive instruments.
Effect of bacterial flora on staphylococcal colonisation of the newborn.
Authors: Speck W T , Driscoll J M , Polin R A , Rosenkranz H S
Published: June 2007

The umbilical and nasopharyngeal flora of newborn infants was examined on days 3, 14, and 42 of life. An analysis of the bacteriological findings suggests that colonisation by either Staphylococcus aureus or Staph. epidermidis prevents colonisation by the other staphylococcus. Similarly, colonisation by Gram-negative bacteria prevents colonisation by staphylococci. Further, this bacterial interference lasts for as long as 42 days, which suggests the possibility of artificially colonising newborns with nonpathogens to prevent subsequent colonisation and disease by virulent microorganisms.
Serum standards for the bioassay of aminoglycosides in cerebrospinal fluid.
Authors: Hammerberg S , Sinai R , Marks M I
Published: June 2007

Four diluents were compared as reference standards for the assay of gentamicin in cerebrospinal fluid (CSF): human CSF, human serum, distilled water, and 150 mmol NaCl/4.5 mmol CaCl2. Standards prepared in pooled human serum were the best alternative to CSF for the assay of gentamicin and were also useful for the assay of tobramycin, netilmicin, amikacin, and sisomicin. The pH (6.0-9.8) of CSF did not alter the results of the assay.
Paraganglioneuroma of the duodenum: an evolutionary hybrid?
Authors: Cooney T , Sweeney E C
Published: June 2007

On light microscopy the neural component of a hybrid neural and endocrine duodenal tumour was characterised by ganglion cells and the endocrine component by epithelioid cells which were both argentaffin and argyrophil. Spindle-shaped cells of indeterminant lineage were also present. Electron microscopy revealed 'pale' and 'dark' cells, the former corresponding to the epithelioid cells and containing many membrane-bound neurosecretory granules, the latter probably corresponding to the spindle cells and containing large numbers of neurofilaments. Occasional cells contained both neurofilaments and secretory granules arranged in a consistent pattern in the cytoplasm. These composite cells may have represented an incompletely differentiated precursor of both the neural and endocrine elements of the tumour.
Congenital syphilis in aborted second trimester fetus: diagnosis by histological study.
Authors: Braunstein H
Published: June 2007

A case of congenital syphilis in an aborted fetus in the second trimester of pregnancy was documented by demonstration of spirochaetes in the fetal liver, despite the fact that diagnostic studies had not been performed on the mother. Suspicion was aroused by the gross and microscopic appearance of the placenta, which appeared identical with the described in proven cases of congenital syphilis. Although these changes have been alleged to be non-specific, the rarity with which such well-developed lesions are encountered in placentas in other circumstances dictates the necessity for further study when they are observed.
Internal sphincter and haemorrhoids: a pathological study.
Authors: Haqqani M T , Hancock B D
Published: June 2007

Histological examination of the lower margin of the internal sphincter has demonstrated an increased amount of fibrous tissue in patients with haemorrhoids compared with controls. This finding may be of relevance to the treatment of haemorrhoids by maximal anal dilatation.
Ethanol formed from arabinose: a rapid method for detecting Escherichia coli.
Authors: Coloe P J
Published: June 2007

Head-space gas-liquid chromatography showed that Escherichia col are consistent in their ability to ferment arabinose with the production of ethanol. In both still and shaken cultures ethanol production is closely aligned with growth. Also ethanol production is related to inoculum size so that varying sizes of inocula may be distinguished by choosing a time of sampling early in growth. The method is suitable as a rapid test for E. coli and is applicable for use in detecting E. coli urinary tract infections.
Head-space gas liquid chromatography for rapid detection of Escherichia coli and Proteus mirabilis in urine.
Authors: Coloe P J
Published: June 2007

Measuring ethanol formed from arabinose by Escherichia coli and dimethyl disulphide formed from methionine by Proteus mirabilis is a rapid way of detecting these organisms in the urine. Ethanol and dimethyl disulphide are detected by head-space gas-liquid chromatography. Since sedimented organisms and not whole urine are used the method is not subject to interference from compounds within the urine. When it was tested on 122 samples of urine, 94 from patients with suspected urinary infection and 28 supposedly uninfected control specimens, the results correlated well with viable counts by a conventional bacteriological method. As well as being rapid (results available in four hours) the method is reliable, easy to use, and could be automated. It now requires extensive testing in a hospital laboratory.
Fluorimetric assay of mycobacterial group-specific hydrolase enzymes.
Authors: Grange J M
Published: June 2007

Fluorigenic substrates prepared from 4-methylumbelliferone provide a simple, convenient method for detecting and assaying group-specific hydrolase activity in small quantities of whole mycobacteria. Physico-chemical properties of the enzymes such as pH dependence and heat stability may also be studied. A technique is described for studying glycosidase, hexosaminidase, acid phosphatase, and acyl esterase activity in seven mycobacterial species. Such techniques will be a useful aid to the taxonomy, identification, and quantification of mycobacteria, especially slowly growing strains in which biochemical properties are very difficult to detect by other techniques.
Chemical and kinetic study of the lipoproteins in abetalipoproteinaemic plasma.
Authors: Shepherd J , Caslake M , Farish E , Fleck A
Published: June 2007

Abetalipoproteinaemic plasma lipoproteins were fractionated by molecular sieve chromatography into two classes on the basis of size. Each class had the same chemical and immunochemical composition and seemed to be interconvertible in vitro, presumably as a result of aggregation/disaggregation. The low levels of circulating apolipoprotein A-I found in abetalipoproteinaemic subjects have been shown by kinetic analysis to result from reduced synthesis of the apoprotein and not from increased catabolism or redistribution between vascular and extravascular compartments.
Grading of astrocytomas using a Quantimet 720 image-analysing computer.
Authors: Robertson A J , Anderson J M , Brown R A , Slidders W , Beck J S
Published: June 2007

Cerebral astrocytomas and undifferentiated gliomas were graded by (1) Kernohan classification on the basis of visual microscopy, and (2) optical density/area profiles obtained by automated densitometry with a Quantimet 720 image-analysing computer. In a retrospective study of 44 patients the Quantimet measurements seemed to give a more accurate assessment of prognosis than visual grading.
Echinococcus granulosus:specific quantification of the two most immunoreactive antigens in hydatid fluids
Authors: Musiani P. , Piantelli M. , Lauriola L. , Arru E. , Pozzuoli R.
Published: June 2007

Preparations of the two most immunoreactiveEchinococcus granulosusantigens (antigens 4 and 5) from sheep hydatid fluid, purified by a simplified method, and monospecific antisera against antigens 4 and 5, prepared by a new procedure, were used to measure the antigenic concentrations of antigens 4 and 5 in swine, sheep, and human hydatid fluids from pulmonary or hepatic cysts. Two bovine samples and two commercial preparations were also tested. The concentration of both antigens was significantly higher in sheep and human hydatid fluids than in swine hydatid fluid. The antigenic content of the two bovine samples and of the two commercial preparations was below the sensitivity level of the method employed. Independently of the species tested, the amount ofEchinococcusantigens was greater in hepatic than in pulmonary cysts. The ratio between the concentrations of antigens 4 and 5 was constant at about 1:10 in the samples from various organs and from different species. When there were enough samples for statistical analysis a linear correlation was found between the contents of these two antigenic components but there was none between the amounts of proteins and the antigenic concentrations in the single cysts. Sheep hydatid fluid must therefore be considered the best source of antigenic material for diagnostic purposes even though in human cysts the antigenic fraction is less contaminated by serum proteins. We describe a reliable method of standardising antigenic material for the immunodiagnosis of hydatid disease.
Comparison of radial haemolysis with haemagglutination inhibition in estimating rubella antibody.
Authors: Appleton P N , Macrae A D
Published: June 2007

Radial haemolysis (RH) for assaying rubella antibody was studied in parallel tests with haemagglutination-inhibition (HAI) on 1102 sera from adult women. Simple to perform and economical in reagents, the RH test is as sensitive as the HAI test and is a useful method of screening antenatal and other sera for rubella immunity. It detects IgG but not IgM antibody.
Cytological basis of histological typing of diffuse Hodgkin's disease. Demonstration of an implied misnomer in the terminology of the Rye classification.
Authors: Livesey A E , Sutherland F I , Brown R A , Beck J S , Macgillivray J B , Slidders W
Published: June 2007

Differential cell counts were made on nine lymph nodes whose structure was replaced by diffuse Hodgkin's disease; two of these nodes had the classical histological appearance of the lymphocytic predominance subtype, four of the mixed cellularity subtype, and three of the lymphocytic depletion subtype. Our attempts to achieve valid sampling methods are recorded. The counts, in general, confirm the postulated histological basis of the Rye classification of the subtypes of the diffuse disease. The major discrepancy is that, contrary to the histological descriptions, our direct counts have shown that lymphocytes, are, in general, more numerous in the lymphocytic depletion than in the mixed cellularity subtypes. The cell counts also show that normal mononuclear cells (mainly fibroblasts and macrophage-type cells) are much more numerous in the mixed cellularity subtype than in the other forms of diffuse Hodgkin's disease; this feature has not been emphasised in the Rye classification. On the basis of our differential counts, a hypothesis is proposed that could explain the natural history of the different subtypes of diffuse Hodgkin's disease as the resultant of three processes: (a) tumour aggressiveness, (b) specific cell-mediated immunological reactions, and (c) non-immunological stromal responses.
Tamm-Horsfall protein in the glomerular capsular space.
Authors: McGiven A R , Hunt J S , Day W A , Bailey R R
Published: June 2007

Tamm-Horsfall protein was detected within the capsular space by immunofluorescence in 7 of 72 consecutive patients on whom renal immunopathological studies were performed. Three patients showed prominent aggregates or crescentic collections affecting 30-50% glomeruli; the remaining four patients showed smaller aggregates between lobules. All patients showed pathological evidence of tubulointerstitial disease. It is suggested that Tamm-Horsfall protein in the capsular space is a sign of intratubular urinary backflow and that Tamm-Horsfall antiserum is a useful addition to the reagents used in the immunofluorescence study of renal biopsies.
Further characterisation of a spontaneously occurring antibasement membrane antibody.
Authors: Ammann A J , Goodman J R
Published: June 2007

A spontaneously occurring antibasement membrane antibody has been further characterised according to antigenic specificity, immunoglobulin class, and tissue localisation using immunofluorescent and immunoelectron microscopic techniques. The autoantibody reacted with the basement membrane of kidney tubules, Bowman's capsule, and the epithelial portion of ileum but not with the basement membrane of skin, cornea, glomerulus, or oesophagus. It also reacted with bile canaliculi, sarcolemmal sheath, and salivary duct. On electron microscopy the antibody was distributed along the basement membrane of Bowman's capsule and renal tubules. Some reactivity against collagen was observed. Antibody activity was found in both IgG and IgM fractions. In immunodeficiency disorders, the autoantibody was found only in patients with selective IgA deficiency.
Specific and non-specific folate binding protein in normal and malignant human tissues
Authors: Corrocher R. , De Sandre G. , Ambrosetti A. , Pachor M. L. , Bambara L. M. , Hoffbrand A. V.
Published: June 2007

Binding of tritiated folic acid by supernatants prepared from extracts of normal and leukaemic leucocytes, normal mucosa, and malignant tumours from different parts of the gastrointestinal tract has been measured using Sephadex-gel filtration and albumin-coated charcoal techniques. Non-specific binding (measured by Sephadex G-75 gel filtration) was almost invariably greater than specific binding measured by albumin-coated charcoal separation of bound and unbound folate. In nine normal leucocyte extracts, binding measured by Sephadex G-75 filtration ranged from 1·3 to 18·2 (mean 8·2) pg/mg protein and by albumin-coated charcoal from 1·0 to 14·8 (mean 6·7) pg/mg protein. Raised specific binding was found in the extracts from leucocytes of eight of 14 patients with chronic granulocytic leukaemia, in four substantially so (389, 121, 108, 59·7 pg/mg protein), but was only marginally increased in one of eight cases of acute myeloid leukaemia and in two of five cases of chronic lymphocytic leukaemia. Binding was normal in the extracts of all three cases of acute lymphoblastic leukaemia tested.

Among the tissues of the gastrointestinal tract binding was greatest by the duodenal mucosa and liver. Extracts of carcinoma of the stomach and colon bound greater amounts of3H-folic acid than the corresponding normal mucosal extracts but the differences were not large. Sephadex G-200 gel chromatography showed more than one binding peak in the extracts of liver and duodenum but only one peak in the other tissues of the gastrointestinal tract, and only one peak, of molecular weight either about 50 000 or over 200 000, in the leucocyte extracts.
Urinary free corticosteroid excretion and renal function.
Authors: Gilliland J , Phillips P J
Published: June 2007

Theoretically urinary free corticosteroid excretion should be affected by renal function and this would make it a less sensitive index of hypercortisolaemia. In 28 consecutive urine samples there was a clear relationship (r = 0.83; P less than 0.001) over a range of creatinine clearances 0.3-200 ml/min. Although an allowance could be made for renal function this would not necessarily improve the discrimination of normal from abnormal. Until data comparing corrected to uncorrected urinary free corticosteroid excretion become available, we recommend a short dexamethasone test as the initial investigation in patients with suspected hypercortisolaemia and abnormal plasma creatinine concentrations.
Evaluation of some methods for the laboratory examination of sputum.
Authors: Tebbutt G M , Coleman D J
Published: June 2007

Methods for the quantitation of leucocytes, squamous epithelial cells, and potential pathogens in sputa are described. Microscopic examination showed that 58% of sputa tested (554/957) were purulent or moderately purulent and 48% were moderately or heavily contaminated by squamous epithelial cells. The presence of squamous epithelial cells indicated oropharyngeal contamination. A simple dilution technique was chosen to compare the isolation of potential pathogens from direct cultures and from dilutions of sputa (10(-7) per ml original sputum). The dilution technique permitted easier reading of sputum cultures and avoided the possible over-reporting of enterobacteria and Pseudomonas aeruginosa, which were frequently found on direct examination. Enterobacteria and Pseudomonas aeruginosa were more likely to be isolated from sputa moderately or heavily contaminated with squamous epithelial cells. The use of counter-current immunoelectrophoresis (CIE) and co-agglutination tests to detect pneumococcal antigen in sputa is described. The presence of antigen in sputum was a more reliable index of lower respiratory tract infection than a positive culture. Co-agglutination tests were simpler to perform and used smaller amounts of expensive antiserum than CIE.
Bone marrow cellularity assessed by point-counting.
Authors: Ho-Yen D O , Slidders W
Published: June 2007

A simple method for rapid point-counting of bone marrow biopsies is described; the method gave an accurate assessment of cellularity in 100 aspiration biopsy specimens. Measurements of marrow cellularity by the visual point-counting technique correlated well with those obtained with a Quantimet 720 image-analysing computer.
Pathology of five Scottish deaths from pneumonic illnesses acquired in Spain due to Legionnaires' disease agent.
Authors: Boyd J F , Buchanan W M , MacLeod T I , Dunn R I , Weir W P
Published: June 2007

The pathology and histology are reported of five Scots who died of severe pneumonic illnesses after holidays in Spain, three in 1973 and two in 1977. There is strong evidence in favour of all the deaths having been due to the newly discovered Legionnaires' disease (LD) agent. The agent (or its soluble antigen) has been visualised in sections of lung tissue by fluorescent-antibody tests in all cases, and the agent has been identified by the Dieterle silver staining method in small numbers in all cases. Serological testing was possible in three of the patients, and two had very high antibody titres against the LD agent. Apart from the extensive and severe nature of the pathological process there is no feature to distinguish pulmonary infection by this agent from that due to more commonly known bacteria capable of causing lobar pneumonia. The severity and extensive nature of the process is partly a reflection of neglect in seeking treatment until late in the infection, and partly a reflection, as revealed in retrospect, on the use of the wrong antibiotic combination during treatment. Erythromycin has been recommended by other workers as the drug of choice against the LD agent. Infection by this organism is not confined to the USA or to Spain and is indigenous also in the United Kingdom.
A comparison of complement fixation, indirect immunofluorescence for viral late antigens, and anti-complement immunofluorescence tests for the detection of cytomegalovirus specific serum antibodies.
Authors: Griffiths P D , Buie K J , Heath R B
Published: June 2007

It was shown that, on average, the titres of the sera obtained by both the anti-complement immunofluorescence (ACIF) and indirect immunofluorescence for viral late antigens (IFA-LA) procedures were approximately eight times higher than those obtained by complement fixation. There was no significant difference between the mean ACIF and IFA-LA titres, but the former method was preferred because it was not affected by non-specific staining. When 406 sera were screened for the presence or absence of antibodies, only five sera (1.2%) gave discordant results with the three tests. None of the tests could detect all of the sera that contained specific antibodies while each test detected over 98% of these sera. Thus, the increased sensitivity of the immunofluorescence procedures did not result in an appreciably increased rate of detection of cytomegalovirus seropositive sera.
Prevalence of pathogenic yeasts and humoral antibodies to candida in diabetic patients.
Authors: Odds F C , Evans E G , Taylor M A , Wales J K
Published: June 2007

The prevalence of oral yeasts and humoral precipitating antibodies to candida was estimated in 204 unselected diabetic patients (172 outpatients and 32 inpatients). Yeasts, mainly Candida albicans, were isolated from the mouths of 41% of the outpatients and precipitins were found in 17.5% although none of the patients had clinically overt candidiasis. The extent of oral yeast colonisation and incidence of antibodies was not related to their antidiabetic treatment or to the duration of their diabetes. It was, however, related to the blood glucose and urine sugar levels at the time they were sampled, the highest incidence being among the diabetic inpatients with high blood glucose levels at the time of sampling and the lowest among outpatients with normal blood glucose levels at the time of sampling. There was no such correlation when diabetic control over the previous 12-month period was considered.
R-factor mediated trimethoprim resistance: result of two three-month clinical surveys.
Authors: Amyes S G , Emmerson A M , Smith J T
Published: June 2007

All urinary tract isolates were monitored in the Whittington Hospital, London for trimethoprim resistance over a three-month period in 1975; this survey was repeated 18 months later in 1977. In the later survey the incidence of trimethoprim resistance had increased significantly, and the proportion of strains carrying R-factors conferring trimethoprim resistance had nearly doubled. The pattern of resistances associated with R-factor trimethoprim resistance also changed betweeen these two surveys.
An unstable haemoglobin, Hb Tacoma beta30 (B12) arg leads to ser, detected at birth by the demonstration of red cell inclusions.
Authors: Deacon-Smith R A , Lee-Potter J P
Published: June 2007

Incomplete expression of human haemoglobin beta-chains at birth may lead to difficulty in the early demonstration of an inherited beta-chain variant. In this case, the rare unstable variant, Hb Tacoma beta30 (B12) arg leads to ser, although not present in cord blood in sufficient amounts to be easily detected by routine electrophoretic techniques, was readily shown to be present by the striking inclusions provoked by prolonged incubation of the neonatal red cells with new methylene blue.
Determination of antithrombin activity by an amidolytic and a clotting procedure.
Authors: Frigola A , Angeloni S , Cerqueti A R
Published: June 2007

Plasma antithrombin activity was measured using an amidolytic method (substrate Chromozym TH) and a clotting method. The mean antithrombin values found in 76 hospital outpatients were 9.4 micronmol/min/ml with the amidolytic procedure and 100.1% of antithrombin activity with the clotting procedure. The two methods correlate fairly well (r = 0.85, P less than 0.01) and show satisfactory reproducibility. Coefficients of variation of 5.9% and 8.8% were obtained respectively with the amidolytic and the clotting procedures. In the presence of very high levels of fibrinogen degradation products, falsely elevated antithrombin activity levels were observed with the clotting procedure but the amidolytic method is essentially unaffected. It is concluded that both methods are suitable for determining antithrombin activity but a well-standardised amidolytic procedure has advantages.
Infection of total hip prostheses by Peptococcus magnus: an immunofluorescence and ELISA study of two cases.
Authors: Taylor A G , Fincham W J , Golding M A , Cook J
Published: June 2007

In two cases of infected total hip replacements, Peptococcus magnus was isolated in pure culture from the implant when it was removed. Fluorescent antibody and ELISA studies have shown that both patients developed an antibody response to this anaerobic coccus soon after the replacement operation. These results suggest that the organism is a true infective agent, which was probably responsible for the failure of the arthroplasty operation.
Use of low ionic strength saline for crossmatching and antibody screening.
Authors: Lown J A , Barr A L , Davis R E
Published: June 2007

A totally revised crossmatching and antibody screening procedure has been designed using low ionic strength saline as the suspending medium for red cells in the saline and enzyme phases as well as the antiglobulin test. The replacement of Pasteur pipettes with precision samplers for all serum and red cell dispensing has resulted in improvement in the reproducibility and standardisation of technique. The major advantages are improved sensitivity, shortened incubation time, and simplicity of technical performance.
Ribosome lamella complex in neoplastic cells of a Sézary's syndrome.
Authors: Sébahoun G , Bayle J , Muratore R , Carcassonne Y
Published: June 2007

The first case of ribosome lamella complex (RLC) is reported in abnormal cells of a Sézary's syndrome, a T cell malignancy. Until now this ultrastructural cytoplasmic inclusion has usually been described in hairy cell leukaemia and other lymphoproliferative syndromes of B cell origin. Since RLC are also observed in abnormal lymphoid T cells, in non lymphoid cells, and moreover in non haematopoietic cells, they lack diagnostic specificity.
Interference of cefoxitin in the creatinine estimation and its clinical relevance.
Authors: Durham S R , Bignell A H , Wise R
Published: June 2007

Serum creatinine and creatinine clearance were measured in 10 healthy volunteers before and after an intravenous injection of 2 g cefoxitin sodium. Results were compared with in vitro work which demonstrated a positive interference by cefoxitin in the Jaffé reaction, the routine laboratory method of creatinine measurement. The serum creatinine estimation should be delayed until at least 2 hours after cefoxitin administration. The creatinine clearance may appear to be falsely high.
Early detection of Chlamydia trachomatis using fluorescent, DNA binding dyes.
Authors: Salari S H , Ward M E
Published: June 2007

HeLa 229 cells were infected with genital tract strains of Chlamydia trachomatis. After incubation for varying times the infected cells were fixed and stained with the fluorescent DNA binding dyes Hoechst 33258 or DAPI for comparison with conventional Giemsa stain. Fluorochrome-treated preparations were examined by incident ultraviolet fluorescence microscopy and the Giemsa-stained preparations by dark-ground light microscopy. Chlamydial inclusion bodies could be identified unambiguously as early as 18 hours after infection of HeLa 229 cells using either Hoechst 33258 or DAPI but not until some 48 hours in Giemsa-stained preparations. The DNA rich chlamydial elementary bodies in infected egg yolk suspension were readily detected using Hoechst 33258. The fluorescent dye technique was simpler and more rapid than Giemsa staining. Using Hoechst 33258 it is possible to speed up the identification of chlamydial isolates growing in tissue culture.
A sensitive single reverse passive haemagglutination test for detecting both HBsAg and anti-HBs.
Authors: Barbara J A , Harrison P J , Howell D R , Cleghorn T E , Dane D S , Briggs M , Cameron C H
Published: June 2007

A trial of a modified reverse passive haemagglutination test for HBsAg using a 0.1% cell suspension instead of the recommended 1% showed an approximately eight-fold increase in detection sensitivity. The test can be performed within 30 minutes and lends itself to mass screening techniques. Confirmation tests can be done using the 0.1% method. In addition, the same serological plates and cells used for HBsAg screening can then be used to screen for high-titre anti-HBs. This makes the overall screening for both HBsAg and high-titre anti-HBs donors cheap and convenient.
Ethylene oxide sterilisation--is it safe?
Authors: Gillespie E H , Jackson J M , Owen G R
Published: June 2007

Tests show that ethylene oxide penetrates and can sterilise long narrow tubes in a hospital ethylene oxide steriliser. Residual ethylene oxide levels in plastic tubing after sterilisation have been estimated. Although initially the levels were very high, storage for four days at room temperature reduced them to a safe level. If adequate controls of the sterilising process and storage are carried out, sterilisation by ethylene oxide is considered to be safe for new plastics and clean equipment.
Pseudo-leptospires in blood culture.
Authors: Rahman M , Macis F R
Published: June 2007

Spiral filaments seen in the blood cultures of two patients with fever and jaundice were initially thought to be leptospires; these were later proved to be artefacts. An investigation was carried out to exclude the possibility of laboratory contamination of the culture media and to find out how these objects were produced. The significance of the findings is discussed in relation to the possibility of a mistaken diagnosis in routine laboratories which have a limited experience of leptospires.
Grouping of streptococci by Streptex.
Authors: Waitkins S A , Ratcliffe J G , Anderson R D , Roberts C
Published: June 2007

Streptex was compared to routine laboratory identification methods available. The results from Streptex sometimes required several attempts before final identification could be achieved. In the main, group D streptococci other than Strep. faecalis failed to group with the Streptex antisera, and this method cannot therefore be used exclusively as a means of identifying this group of streptococci.
Urinary total porphyrins by ion exchange analysis: reference values for the normal range and remarks on preformed prophyrins in acute porphyria urine.
Authors: Fogstrup J , With T K
Published: June 2007

Determination of total porphyrin with a rapid and east method (ion exchange; spectrophotometry) was performed on 57 morning urine samples from laboratory personnel, 59 arbitrary day urine samples from blood donors, and 90 24-hour urines from medical inpatients. The upper reference limit for morning urine was 0.32 mumol/l but even a value as high as 0.7 mumol/l was found. In the donor urines the upper limit was 0.155 mumol/l. The 24-hour urines showed an upper reference limit of 271 microgram/24 h. These values are in good agreement with values from the literature, mostly based on extraction analyses. Tracings of the absorption curves in the region of 380--430 nm were performed in all analyses and showed that the non-porphyrin absorption was close to linear in most cases. Studies of porphyric urines gave no support to the claim that preformed porphyrins not formed from porphobilinogen are excreted in this disease.
An evaluation of simultaneously administered free and intrinsic factor bound radioactive cyanocobalamin in the diagnosis of pernicious anaemia in the elderly
Authors: Pathy M. S. , Kirkman S. , Molloy M. J.
Published: June 2007

The definitive diagnosis of pernicious anaemia (PA) in the elderly is by no means always straightforward, particularly when inappropriate medication has been introduced before the institution of specific investigatory procedures. A detailed haematological study was carried out on 301 patients aged 60-95 with a serum B12concentration at the laboratory's lower level of normal of 150 ng per litre (Euglena gracilisassay).

The diagnosis of PA was based on strict predetermined haematological criteria. All patients were subsequently studied by the simultaneous oral administration of the dual isotopes57Co-labelled B12bound to intrinsic factor and free58Co-labelled B12(Dicopac test), and urine was collected over 24 hours after an intramuscular dose of 1 mg nonradioactive B12for estimation of the57Co/58Co B12ratio; 255 patients satisfied all criteria for final analysis.

The Radiochemical Centre, Amersham suggests an upper limit of the normal range for the57Co/58Co ratio of 1·3 with a lower limit for PA of 2·0. We were unable to show a sharp borderline in the57Co/58Co B12ratio between those patients shown by other criteria to have PA and those who do not have PA; 34% of the 71 established patients had a ratio below 2·0. From our series a ratio borderline drawn at 1·4 gave only one false negative (1·4% of the PA group). Of the 175 non-PA cases, nine (5%) gave false positive results; four of these had58Co excretion levels high enough to make misdiagnosis unlikely.

In a proportion of patients the57Co/58Co B12ratio was estimated at regular intervals for 36-hour periods. Maximum accuracy of isotope measurement on a single specimen was obtained 8-20 hours after isotope dosing.

The Dicopac investigation is a useful simple screening test in the differential diagnosis of patients with a megaloblastic bone marrow and combined low serum B12and folate concentrations. When carried out by the standard technique, the degree of discrimination between normal and abnormal ratios is of limited diagnostic significance in one-third of patients.
Toxocariasis: serological diagnosis by enzyme immunoassay.
Authors: de Savigny D H , Voller A , Woodruff A W
Published: June 2007

An enzyme-immunoassay was developed to measure the concentration of serum antibody specific for the secretory antigens released by migrating toxocaral larvae. This technique was evaluated by testing sera from healthy UK adults, and from patients with and without toxocariasis. In 922 healthy adults, 2.6% were found to have elevated specific antibody levels. Elevated values were observed twice as frequently in males as in females but showed no significant regression with age between 20 and 65 years. Of 62 patients with non-toxocaral helminthic infections, all had antitoxocaral antibody levels within the range of values observed in healthy controls and had a mean level which was not significantly elevated. All of 13 patients with clinical toxocariasis had enzyme-linked immunosorbent assay (ELISA) antibody levels above the 100th percentiles of both the healthy population and the helminth-infected group and had a significantly high mean value (p less than 0.001) more than 12 times that of the healthy or infected controls. The high degree of sensitivity and specificity of the toxocariasis enzyme-immunoassay indicates that this new test should be useful in reference immunodiagnostic applications and in large-scale seroepidemiological surveys.