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Biomedical subjects

K Hyde

Publications and source records attributed to K Hyde.

At least 19 recordsLinked to original sources
PubMed1995

Immunostaining of whole agar cultures by APAAP.

We have adapted the alkaline phosphatase-anti alkaline phosphatase (APAAP) technique to demonstrate cell antigen distributions in intact agar culture. The method facilitates batch processing and is no less convenient to perform than standard APAAP procedures. Myeloid and lymphoid antigens generally demonstrated strong staining intensity. However, staining at day 0 consistently produced no antigen expression for two monoclonals (CD11c and CD34) in contrast to positivity in parallel cytospins. CD11c showed rapidly increasing antigen expression over subsequent days of culture whereas the expression of CD34 could not be shown in conventional agar culture at any time from day 0 to day 14. Positivity was only restored in CD34-positive leukaemic cells using a modified culture technique in which cells were cultured as pre-formed small aggregates. Assessment of these aggregates extended to cell cycle analysis using anti-bromodeoxyuridine. CD71 positivity in normal culture samples correlated with colony configuration (whether clones were 'spread' or 'tight' in appearance). CD38 staining of normal bone marrow culture at day 7 showed asymmetrical staining of cells in a small number of micro-groups. The clonal detection of aberrant antigens (CD7, CD2) for assessment of minimal residual disease in AML was a disappointment due to the relative frequency of positive clones in normal culture.

Agar↗
PubMed1992

Two-colour immunoenzymatic technique using sequential staining by APAAP to evaluate two cell antigens.

AIMS: To extend the alkaline phosphatase-antialkaline phosphatase (APAAP) immunoenzyme single stain method to a more generally applicable double stain technique. This will allow two primary antibodies of the same isotype of IgG and specifically the nuclear antigen bromodeoxyuridine (BRdU) to be evaluated with a cell surface antigen identifier. METHOD: Sequential applications of the APAAP method showed two antigen sites by different dye couplings to a common alkaline phosphatase substrate, producing blue and red reaction products on the same slide. Antigens on different cell populations as well as those in different compartments of the same cell were analysed. The method allowed a surface antigen monoclonal to be revealed first, using an optimal fixative, before alcohol/gluteraldehyde fixation was used to start the second (BRdU) staining sequence. RESULTS: An analysis of double staining of T lymphocyte subsets (CD4 and CD8) showed no significant difference in the order of application of the primaries (n = 10) and no significant difference from their corresponding single stain results (n = 50), confirming the validity of the technique where antigens are exclusively distributed. Other examples, including antigens distributed in different compartments of the same cell, displayed discrete staining which implied validity. CONCLUSION: Double staining by APAAP with this technique seems to be applicable to those cases where antigens are exclusively distributed and includes cases where different compartments of the same cell are stained. It is especially useful in revealing antigens that require different fixation and preparation--that is DNA incorporated BRdU with a surface antigen. But it does seem to have a limited ability to produce a dual colour at a common site.

Antibodies, Monoclonal↗
PubMed1992

A computerized database for haemophilia patient records.

A microcomputer database system for the storage, retrieval, and statistical analysis of data associated with the treatment of haemophilia and other defects of haemostasis is described. The hardware requirements are an IBM compatible PC with both hard and floppy disc drives and a suitable printer. The system was written using Smartware II, a powerful integrated software package which incorporates database, word processor, spreadsheet and communications functions. The programs were written with flexibility in mind and can be readily adapted to accommodate the work patterns of any haemophilia centre. This system has now been operational in the Regional Haemophilia Centre at the Manchester Royal Infirmary since January 1990. Its introduction has led to a marked improvement in the efficiency of patient data handling with significant savings in staff time.

England↗
PubMed1991

Continuous assessment of performance in External Quality Assessment Schemes.

A simple, standardised procedure was used for processing data from External Quality Assessment Schemes and for producing performance charts. A graphic presentation of cumulative performance is generated for each test, using an integrated software package to display results in a standardised format. This permits interpretation of satisfactory and poor performances and facilitates identification of "outlying" results. Although most results are recorded as Deviation Indices, the system is flexible and can be used to present other performance variables, such as Percentage Deviations and relevant textual information relating to interpretative assessments. It is therefore readily adaptable for use by other pathology disciplines.

Data Interpretation, Statistical↗
PubMed1991

Comparative ferrokinetic study with initial and extended iron clearance models.

Erythrokinetic studies were performed on 10 patients with chronic myelofibrosis and 11 patients with myelodysplasia (MDS). Values for plasma iron turnover, marrow iron turnover, and erythron transferrin uptake were derived using two ferrokinetic models. One entailed analysis of the extended plasma iron clearance over a number of days, the other comprised analysis of the initial plasma iron clearance during the first few hours of the study. A close correlation was found between the variables quantifying total erythropoiesis (marrow iron turnover and erythron transferrin uptake) in the two methodologies. Functional classifications produced by both models and based on the values for plasma iron turnover, marrow iron turnover, and erythron transferrin uptake were compared. Both models identified functional heterogeneity in the group with myelofibrosis and functional homogeneity within the MDS group. Each method produced comparable data on erythropoiesis. The main reason for analysing the extended plasma iron clearance is to differentiate levels of effective and ineffective erythropoiesis. The short analysis presents the practical advantages associated with a one-day study. This could be further enhanced if the level of effective erythropoiesis could be clearly defined.

Bone Marrow↗
PubMed1989

Success of the Fasanella-Servat operation independent of Müller's smooth muscle excision.

In an attempt to elucidate the mechanism whereby the Fasanella-Servat operation corrects ptosis, the authors examined the histopathologic features of 40 consecutive surgical specimens from 37 patients. Because all specimens contained tarsus, this tissue was graded into two groups according to vertical height: (1) minimal (30%) and (2) moderate (70%). Müller's smooth muscle was graded into four groups: (1) absent to negligible (42.5%); (2) minimal (45%); (3) moderate (10%); and (4) large (2.5%). Levator aponeurosis was absent, and conjunctive was present, in all resections. Accessory lacrimal gland tissue was present in 42.5% of cases and did not cause decreased tear production. Although 87.5% of cases had absent to minimal smooth muscle resections, these patients had equally successful results in comparison to patients with moderate to large amounts of smooth muscle resections. Based on these data, the authors have concluded that the effectiveness of the Fasanella-Servat operation does not depend on a Müllerectomy, but instead is probably due to a combination of other factors: (1) a vertical posterior lamellar shortening; (2) secondary contractile cicatrization of the wound; and (3) plication or advancement of the Müller's smooth muscle-levator aponeurosis complex on the tarsus.

Adolescent↗
PubMed1989

Detection of erythroid hypoplasia in myelofibrosis using erythrokinetic studies.

The iron kinetic model described by Ricketts et al was used to study haemopoiesis in chronic myelofibrosis. The clearance of 59Fe-labelled transferrin from the plasma was analysed to quantify total, effective, and ineffective erythropoiesis, denoted by the terms marrow iron turnover (MIT), red cell iron turnover (RCIT), and per cent ineffective iron turnover (IIT%), respectively, in 12 cases of this disease. The patterns obtained were variable: values for MIT ranged from 24.4 to 510 mumol/l blood/day; those for RCIT from 0.4 to 119 mumol/l blood/day; and those for IIT% from 67 to 98%. One noteworthy feature was the presence in two cases of functional erythroid hypoplasia; these were characterised by severely reduced values for MIT (24.4 and 28 mumol/l blood/day) and RCIT (0.4 and 8 mumol/l blood/day.) A systematic study of the erythrokinetic features of myelofibrosis may indicate that erythroid hypoplasia is a more common cause of anaemia in this disease than has been previously recognised.

Anemia, Aplastic↗
PubMed1989

Simple method for granulocyte-macrophage cell culture and staining in soft agar: comparison with a standard research technique.

A modified, small volume, two phase, disc culture system for CFU-GM (seven and 14 days of incubation) was compared with a standard single layer system. The 1 ml single layer cultures were counted unstained in situ before both sets of cultures were transferred to glass slides for staining. Bone marrows were cultured from forty eight subjects, including normal controls and patients with acute non-lymphoblastic leukaemia, acute lymphoblastic leukaemia, and myelodysplastic syndrome. Observer error was least with the disc cultures, whereas variation between replicate cultures was similar for both methods. A high degree of correlation was found between the two methods for both day 7 (r = 0.90) and day 14 (r = 0.91) cultures. The number of colonies and clusters was higher with the disc system, indicating better cloning efficiency. Analysis of subsets of clinical groups showed similar patterns of abnormality with both systems. The simplicity of the method makes the use of this technology possible in most laboratories, and the superior morphological resolution may increase the clinical usefulness of such studies.

Bone Marrow↗
PubMed1988

A proposal for further standardization of red blood cell glucose 6-phosphate dehydrogenase determinations.

A number of factors were identified which influence the determination of red blood cell glucose 6-phosphate dehydrogenase (G6PD). White blood cells were effectively removed from whole blood by filtration, resulting in a mean reduction of G6PD activity of greater than 1.0 U/g Hb. Optimal enzyme activity was found using haemolysates with a Hb concentration in the range 0.4-1.2 g/dl. At a constant temperature of 30 degrees C, maximum linear rate of activity was achieved between the 10th and 20th min after initiation of the reaction. As a result of these findings, modifications to standardize the assay system were introduced. The new method exhibits excellent precision (CV less than 2.0%) with normal samples, and 95% reference ranges of 7.00-11.15 U/g Hb for adult females and 7.05-10.75 U/g Hb for adult males.

Adolescent↗
PubMed1988

Complications of blepharoplasty.

Various involutional eyelid changes, such as fine rhytids, dynamic laugh lines in the lateral canthal area, secondary malar bags, and dermal pigmentation are not effectively treated by a blepharoplasty. Preoperative discussions are important to ensure that adequate expectations are held. Too often, a patient is informed that fine rhytids and dermal pigmentation can be successfully treated with lid surgery alone. The importance of the preoperative interview in determining patient expectations cannot be minimized. Preoperative ophthalmic and periocular examination is essential to identify any pre-existing pathology. Even though the surgical approach is adapted to the individual needs of each patient, the most meticulous of surgeons is predetermined to encounter his or her share of complications. There is a fine line between a "complication" and a common, but unwanted, sequela of surgery. Patients, overwhelmingly, do have some widening of their palpebral fissures, even following conservative surgery. Unfortunately, this occasionally creates ocular irritation or gross discomfort in allergic or borderline dry eyed patients. Lower eyelid bowing is so common even in the face of minimal skin resection that we do not consider it a "complication" if it does not produce a functional problem. It is humbling for all surgeons to review their blepharoplasty problems and helpful to recognize that, statistically, we are all due to face these events eventually.

Eye Hemorrhage↗
PubMed1985

Assessment of the role of an immunofluorescent terminal deoxynucleotidyl transferase kit.

The laboratory use of a commercial kit for the immunofluorescent measurement of terminal deoxynucleotidyl transferase (BRL Terminal Deoxynucleotidyl Transferase Immunofluorescent Assay System) has been investigated. The precision of the assay system with stored patient material and its performance with regard to ease of microscopic examination were assessed. The intensity of the fluorescence obtained was found to vary. However, the precision of the assay system on fixed slides stored at -30 degrees C was satisfactory; the method simple, and results obtainable in a relatively short space of time.

Bone Marrow↗
PubMed1982

Microcomputer data handling for the Phoenix system.

A modified Phoenix data processing system has been introduced into the laboratories at the Manchester Royal Infirmary. Specifications for the design and installation of microcomputers to link automated instruments to this host computer are described. The operation and data manipulation facilities of the microcomputer are included. It is concluded that these devices aid the management of data within the department and improve the quality of service.

Blood Cell Count↗
PubMed1981

Chromium genotoxicity as influenced by complexation and rate effects.

Conclusions as to the mutagenicity and carcinogenicity of metal salts can be ambiguous and misleading, especially for metal ions having a high charge/radius ratio, hence a strong tendency to hydrolyze. Using the rec-assay, we determined whether the mutagenicity of chromium salts was reduced by complexation, as in the case of Cr(VI), or induced in the case of Cr(III). We find that several chelants, in proportion to concentration, reduce or eliminate the mutagenicity of Cr2O32-. These include EDTA, salicylate (SA), and Tiron (disodium 1,2-dihydroxylbenzene-3,5-disulfonate). Cr(III) was rendered slightly mutagenic by salicylate and citrate. None of the chelating agents or their combinations were mutagenic.

Bacillus subtilis↗
PubMed1981

Comparison of liquid and dried sodium citrate as the anticoagulant for Thrombotest and prothrombin time estimations.

Samples which are taken for the Quick one-stage prothrombin time estimation for the control of patients receiving oral anticoagulant treatment are by convention taken into liquid sodium citrate anticoagulant. Dried sodium citrate has been rejected on the grounds that excessive haemolysis causes activation of clotting factors and therefore falsely short clotting times. Tests were performed with both liquid and dried sodium citrate as an anticoagulant for prothrombin time and Thrombotest. No clinically significant difference between the values given by the two anticoagulants was observed.

Anticoagulants↗
PubMed1980

Evaluation of the Hematrak differential leucocyte counter.

The main objectives of the evaluation were to: (1) assess accuracy and precision of cell recognition, (2) establish 'reference ranges' and compare with existing ranges, (3) assess machine reliability and suitability for normal working patterns. The accuracy and precision of the instrument were found to be acceptable, within the limits imposed by counting 100 cells in a wedge smear. Reference ranges compared well with established ranges with the exception of monocytes and eosinophils. the instrument was initially evaluated over a 12-week period and has been in routine use for 18 months durng which time it has been found to be very reliable. It has fitted in with the laboratory working patterns without causing any changes in work flow.

Blood Specimen Collection↗