Risk factors associated with noncompliance with methadone substitution therapy (MST) and relapse among chronic opiate users in an Outer London community.

AIMS: The purpose of this study was to identify the risk factors associated with noncompliance with methadone substitution therapy (MST) and hence relapse in chronic opiate-dependent users, as this has major clinical implications especially in community-based detoxification programmes. BACKGROUND: Community mental health nurses (CMHNs) and other health/social care professionals need to be aware of the main risk factors associated with MST noncompliance among long-term opiate users living within their catchment areas. The sex-matched patterns of biopsychosocial risk factors can be useful predictors of the ability of clients to comply with MST and their likelihood to complete the detoxification programme. A knowledge of the patterns of these high-risk factors also allows the care professionals to: (1) draw-up or re-draw care contracts that reflect their patients' biopsychosocial circumstances; (2) initiate much broader, client-centred, relapse prevention strategies; (3) select suitable patients for specialized detoxification contracts; and (4) modify the care approach from detoxification to maintenance contracting particularly for clients with low predicted scores for the former contract-type. METHODS: As successfully demonstrated in this semi-quantitative descriptive investigation, identification of these sex-typed biopsychosocial high-risk factors can easily be undertaken during assessment and when the clients attend regular review. In this study, numerical information was gathered during personal face-to-face interviews, and was supplemented with that contained in past multidisciplinary case notes. RESULTS: Overall, the medication noncompliant female clients were associated with personality trait, decreased educational expectations, everyday life stresses, ambivalent thoughts, social company availability, comorbidity, boredom, and family-related conflicts; whereas the noncompliant male clients were associated with poor motivation, fashion and reputation, peer association, uncontrollable drug-cravings, drug availability, major life events, too stringent prescribing/poor client-centred care package, heavy intravenous users, young polydrug users, and triple users. CONCLUSIONS: These findings have important nursing practice connotations. This study is advocating the routine identification of biopsychosocial high-risk factors associated with MST noncompliance by all CMHNs working with chronic opiate-dependent users.

Osmium tetroxide-potassium ferrocyanide intensification of a diaminobenzidine product obtained by photoconversion of a fluorescent label: a study of human neutrophil granules.

A diaminobenzidine posttreatment employing osmium tetroxide and potassium ferrocyanide was successfully used to intensify the diaminobenzidine stain formed by photoconversion of immunofluorescent labelling. Lactoferrin labelled granules became visible following the photoconversion process. Adequate diaminobenzidine staining was obtained after irradiating the cytospin preparations with ultraviolet light for 30-40 min. The diaminobenzidine stain had advantages over the fluorescent stain owing to its greater stability, greater density, and ability to be intensified. The enhancement procedure intensified both the color and density of the diaminobenzidine product. Consequently, shorter irradiation times could be used. Osmium tetroxide solutions of 3-4% increased the intensity with minimal background staining. Ultrastructural immunogold cytochemistry on ultrathin sections confirmed the existence of the lactoferrin labelled structures observed by light microscopy of cytospin preparations indicating that these were secondary granules. The photomicroscopy procedures used in this study were simple to perform and could be applied to studies of other cellular antigens prior to using immunoultramicroscopy.

Changes in neutrophil granule protein and cytoplasmic fibrils in human acute myeloid leukemias.

Granule protein deficiencies in morphologically mature neutrophil cells of peripheral blood from human patients with acute myeloid leukemia was demonstrated using post-embedding immunocytochemistry. Abnormal immunoreactivity of granule proteins was detected in seven of nine patients. Decreased immunoreactivity patterns were found more for the primary granule markers elastase and myeloperoxidase than for the secondary granule marker lactoferrin. Leukemias with a predominant myeloid component, in contrast to those with a predominant monocytoid component, had more neutrophil cells showing immunodeficiencies for one or more granule markers. The proportion of neutrophil cells showing immunodeficiencies varied greatly for each granule marker; more variation was obtained for elastase, lactoferrin and myeloperoxidase than for lysozyme, possibly because lysozyme is a marker for both granule types. In addition, no correlation could be found between any of the immunoreactivity deficiencies for the neutrophil granule glycoproteins elastase, lactoferrin, lysozyme and myeloperoxidase and the abundance of a particular set of ultrastructural features in the circulating leukemic cells from any of the nine patients. Nonetheless, most of the immature myeloid cells from peripheral blood of leukemic patients showing neutrophil protein immunoreactivity abnormalities in one or more granule markers often and randomly displayed one or more unusual ultrastructural features. The clinical and pathological significance of neutrophil granule protein deficiencies and the abundance of fibrillar structures in malignant myeloid cells presently is uncertain.

Integration of family planning into maternal and child health and other services at Harare Central Hospital.

PIP: To increase the accessibility, availability, and acceptance of family planning methods and counseling, family planning services were integrated with maternal and child health care services at Harare Central Hospital in Zimbabwe. The Family Planning Project was implemented with hopes that mothers would seek contraceptive methods and counseling concurrent with their or their children's hospital admission, thereby making facility and service inaccessibility a thing of the past. Ante-natal, post-natal, and postabortal women were targeted for project outreach at the facility, along with patients suffering chronic medical and psychiatric problems, and mothers of malnourished children. Weaknesses of family planning provision at the hospital prior to the project are presented in the component parts of pharmacy, emergency gynecology unit, outpatient department, ante-natal clinic, post-partum care, post-natal clinic, and the general hospital. 2 full-time nurse- midwives and 2 part-time gynecologists counsel and provide services for the Family Planning Project. Other programmatic changes and improvements are described. There were 3,822 new acceptors and 5,423 return visits during the 1st project year, with the nurse-midwives providing 3,114 couple-years of protection, equal to 5.1% of the total provided by all 35 national family planning council clinics. Additional results, plans for the future, and problem areas are further discussed. The project, undertaken with few resources and high motivation, yielded high family planning acceptance rates with markedly less inconvenience for acceptors.^ieng

Analysis of human neutrophil granule protein composition in chronic myeloid leukaemia by immuno-electron microscopy.

In this study immuno-electron microscopy was used to assay, semi-quantitatively, the granule contents of elastase, lactoferrin, lysozyme and myeloperoxidase in human peripheral blood neutrophils from 13 chronic myeloid leukaemia patients in the chronic phase of the disease and from normal non-smoking donors. The fixation conditions that adequately preserved the antibody binding capacities of these antigens and reasonably preserved the ultrastructure of the neutrophils were selected by light-microscopic immunoperoxidase cytochemistry on cytospin smears. Immunogold cytochemistry on LR White resin sections localised elastase and myeloperoxidase to the primary granules, lactoferrin to the secondary granules and lysozyme to both types of granule. When applicable, peroxidase cytochemistry was combined with immunogold staining making it easier to distinguish the primary from the secondary granules. A comparison of the immunolabelling density values obtained for the leukaemic and normal states revealed no significant abnormalities in the immunoreactivity patterns for any of these neutrophil granule antigens in the leukaemic patients. All 13 patients gave normal immunostaining reactivities for these neutrophil granule proteins. Consequently the distribution patterns of these proteins, as shown in this study, cannot be used as indices in distinguishing chronic myeloid leukaemic neutrophils from normal neutrophils.

Combination of diaminobenzidine staining and immunogold labeling: a novel technical approach to identify lysozyme in human neutrophil cells.

In this study, a combination of the diaminobenzidine staining procedure for myeloperoxidase and the immunogold labeling technique was successfully used to show that lysozyme is indeed found in both the primary and secondary type granules of human neutrophils. Following the systematic selection of processing conditions by light microscopic peroxidase anti-peroxidase cytochemistry, on slide preparations, consistent gold labeling was obtained over both types of granules. The combination of myeloperoxidase and immunogold cytochemical procedures permitted the lysozyme-labeling pattern of the small-sized granules to be studied in isolation, thereby confirming the existence of lysozyme in secondary granules. In addition, myeloperoxidase was observed in the large-sized, lysozyme-positive, granules by both cytochemical and immunocytochemical methods, thereby confirming that these labeled structures were primary granules. Morphometrical analysis confirmed that there was a significant difference in mean size between the lysozyme-positive, myeloperoxidase-positive, granules and the lysozyme-positive, myeloperoxidase-negative, granules. The former were significantly larger in size than the latter. In conclusion, although the localization of lysozyme in human neutrophils by the immunogold technique is confirmatory, the combination of enzyme cytochemistry and immunocytochemistry is a novel technical approach that permits the lysozyme-labeling patterns of granule types to be studied in isolation. This double labeling technique is relatively straightforward and, as such, consistent immunostaining can be routinely obtained using intact cells.

Applicability of using acrylic resins in post-embedding ultrastructural immunolabelling of human neutrophil granule proteins.

In this study, quantitative assessments were carried out, (1) by light microscopy during tissue preparation for electron microscopy and (2) by electron microscopy after on-grid immunogold staining, to determine the suitability of using LR White and Lowicryl K4M thin sections to identify lactoferrin and elastase in the granules of human neutrophil leucocytes. Quantitative assessment of the effect of fixation, dehydration and embedding on the preservation of antigenicity during tissue preparation for electron microscopy, using light microscopic peroxidase anti-peroxidase immunocytochemistry, enabled the selection of preparation conditions that adequately preserved both antigenicity and ultrastructure. OsO4 post-fixation, following primary aldehyde fixation, improved the retention of antigenicity during dehydration and embedding and the preservation of fine structure. Partial rather than complete dehydration retained more of the antigenicity. The efficiency, sensitivity and resolution of immunolabelling and the ultrastructure and quality of sections achieved after embedding in LR White were superior to those obtained after embedding in Lowicryl K4M. Consequently room temperature embedding in LR White following double fixation and partial dehydration is a better and more reliable preparation technique than low-temperature embedding in Lowicryl K4M following single fixation and partial dehydration for localizing lactoferrin and elastase to the specific and primary granules respectively in human neutrophilic granulocytes by the on-grid immunogold staining method.

Use of light microscopic immunotechniques in selecting preparation conditions and immunoprobes for ultrastructural immunolabelling of lactoferrin.

Successful postembedding immunolabelling for electron microscopy is sometimes difficult to achieve. We propose that light microscopy can be used (1) to detect quickly processing steps which have an adverse effect on the tissue antigenicity and (2) to check the specific reactivity of the immunogold detecting system normally employed at the ultrastructural level. The individual steps of fixation, dehydration and embedding were tested for their ability to preserve antigenicity by light microscopic peroxidase--anti-peroxidase cytochemistry. Steps that severely reduced antigenicity were replaced by less destructive alternatives compatible with reasonable ultrastructural preservation. The specific reactivity of the immunogold detecting system was assessed by using the light microscopic immunogold-silver staining method. We studied the antigen lactoferrin in human neutrophilic granulocytes from patients with chronic myeloid leukaemia. We obtained strong immunolabelling of specific granules and good ultrastructural preservation using routine methods at room temperature. For lactoferrin the method of choice was to fix in 3% paraformaldehyde/0.1% glutaraldehyde followed by 1% OsO4, dehydrate in 70% ethanol, embed in LR White resin and polymerize at 40 degrees C for 40 h. These conditions may not be suitable for all antigens and we emphasize that for each new antigen a similar study should be carried out.