Presence of proliferating (MiB-1-positive) cells in cervical smears of women infected with HIV is associated with clinical outcome: a study of Brazilian women.

The hypothesis tested was that there is an association between the presence of proliferating (MiB-1-positive) cervical cells and clinical outcome of women infected with human immunodeficiency virus (HIV). Female partners (attending the Gynecology Outpatients Clinic of the University Hospital of Rio Grande, Brazil) of known HIV-positive (HIV+) men were used for this pilot study. Among these women, 25 were also HIV+. Papanicolaou smears of these 25 HIV+ women and of 44 HIV- women were graded as negative, CIN I, CIN II, or CIN III, using neural network screening. MiB-1 grading and HPV identification were also performed. The immune status of patients was determined using the current Centers for Disease Control classification. In agreement with the scientific literature, in these Brazilian women both CIN and HPV were associated with HIV. In the HIV+ women, the immune status tends to correlate with MiB-1 grading. Also, in the one case in whom progression from CIN I to invasive cervical carcinoma was observed, the smear contained many MiB-1-positive cells. Staining cervical smears of HIV+ women is a simple procedure to get an indication of clinical outcome of the patient.

Analysis of three-dimensional architecture in Feulgen-stained fine-needle liver aspirates, using confocal scanning laser microscopy.

Early and accurate diagnosis of hepatocellular carcinoma in liver nodules using fine-needle aspiration biopsy can still pose difficulties, particularly in cirrhotic nodules containing dysplastic hepatocytes. Loss of architectural clues, which may aid diagnosis, can be overcome by using confocal scanning laser microscopic examination of tissue fragments in smears without the need for further processing of slides. The basic fuchsin of the Feulgen method resulted in excellent confocal images without the need for further processing of the smears. Clear morphological differences in three-dimensional reconstructions of optically sectioned tissue fragments were demonstrated in normal, hyperplastic, and malignant smears as an aid to diagnosis.

Microwave-stimulated Jones-Marres method for staining fungi in brain tissue of immunocompromised patients.

Life-threatening fungal infections have increased significantly in the past decade due to the rising number of immunocompromised patients. Serological diagnosis of most fungal infections is unreliable and blood cultures are positive in only 50% of premortem cases; therefore, tissue sampling together with fast, reliable staining of fungi should be carried out to reach the correct, timely diagnosis. We developed, partly serendipitously, a microwave silver staining method for fungi in histological sections. During a differentiation step in periodic acid, background staining is removed. This rapid staining method can be combined with immunostaining, for example, the alpha smooth muscle actin method, to visualize blood vessels. Silver staining results were optimized using the recently developed MicroMED BASIC microwave labstation for histology (Milestone srl, Italy), featuring no-touch temperature measurements and PC control.

T lymphocytes and immunosuppression in the burned patient: a review.

Extensive thermal trauma results in impaired immune function which has been attributed to a reduction in T lymphocyte numbers, increased suppressor cell activity, serum suppressive factors and altered cytokine synthesis and receptor expression on T cells. Numeric and phenotypic changes in T lymphocytes, T cell proliferation and functional responses of T lymphocytes in recent studies using murine models and patients are described.

Childbirth: management of labour through the ages.

This paper examines the management of labour through the ages, and looks particularly at the emergence of invasive techniques in childbirth and the development of pain management. Obstetric practices throughout the 17th and 18th centuries are explored.

Plasminogen activators in normal and malignant oral epithelium in vivo and in vitro.

Urokinase-type (uPA) and tissue-type (tPA) plasminogen activators were identified by fibrinolytic autography in the sulcus epithelium of human gingival mucosa but not in the orthokeratinized gingival epithelium. Fibrinolytic activity was present only over blood vessels in frozen sections of oral squamous cell carcinomas, the malignant epithelial cells showing no plasminogen activator activity. Plasminogen activators could not be demonstrated in either the sulcus or gingival epithelium by immunofluorescence, but both uPA and tPA were found in occasional squamous carcinoma cells. Fibrinolytic activity of culture fluids from epithelial explants grown in vitro from human gingival mucosa showed marked variation, but activity was much higher in the culture supernatants than in the cell lysates. Fibrinolytic activity of culture fluids from epithelial explants of squamous cell carcinomas was low both in supernatants and lysates. Zymogram overlays of sodium dodecyl sulphate-polyacrylamide electrophoretic gels from culture supernatants showed that the low fibrinolytic activity of culture supernatants of oral squamous cell carcinomas was due to the associated presence of plasminogen activator inhibitors. The fibrinolytic activity in the zymogram was due predominantly to uPA but some lysis was due also to tPA.

Pathophysiology of soft tissue repair.

Inflammation with subsequent migration of leucocytes and connective tissue cells to the site of damage, together with the release of cytokines by these cells are essential for healing in common sports injuries. Injury to the musculotendinous unit resulting from either blunt trauma, tears or laceration, heal primarily by formation of granulation tissue and scarring. Early diagnosis with appropriate therapy may minimize any potential loss of function. Ligament repair also follows a classical healing response, although the quality of healing is site dependent and may be related to exposure to synovial fluid. In contrast, cartilage, which is avascular, lacks the inflammatory response seen in other connective tissues and this frequently results in poor tissue repair with subsequent degeneration of the injured cartilage. Mechanisms of repair in these tissues are described.

Tissue reactions induced by hydrocolloid wound dressings.

Porcine full-thickness excisional wounds were treated with 4 different hydrocolloid (HCD) dressings--DuoDERM (ConvaTec/E. R. Squibb), Intrasite HCD (Smith and Nephew Medical), Tegasorb (3M) and Replicare (Smith and Nephew Medical). Animals were killed at 4, 10, 21 and 90 d post-wound, excision sites were fixed in formalin and processed for histological analysis. Granulomatous lesions were observed following treatment with each of the 4 HCD dressings. Such lesions developed between 4 and 10 d post-wound, exhibiting little evidence of resolution at 90 d post-wound. Of the 4 dressings examined, DuoDERM and Intrasite HCD precipitated the most severe reaction, each treatment resulting in granulomata with a distinct and different morphology. Treatment with DuoDERM resulted in granulomata characterised by a random distribution of dendritic cells, epithelioid cells, multinucleated giant cells, lymphocytes and plasma cells. In contrast, treatment with Intrasite HCD resulted in highly organised granulomata, consisting of a central focus of epithelioid cells surrounded by a peripheral cuff of macrophages, lymphocytes and plasma cells. This experimental study highlights chronic inflammatory lesions that may, if reflected in the clinical environment, question the efficacy and indication of HCD dressings in the treatment of wounds having a number of different aetiologies.

Ultrastructural alterations associated with in vivo and in vitro bound pemphigus antibodies in cultured oral epithelial cells.

Pemphigus vulgaris is characterized by bound and circulating IgG antibodies to an epithelial antigen, antibody binding resulting in acantholysis. This study examines the role of pemphigus serum in the initiation and perpetuation of acantholysis. Oral epithelial cells derived from a patient with pemphigus vulgaris were cultured in vitro in the presence of normal serum. Also, normal oral epithelial cells were cultured in pemphigus serum for between 0.5 and 72 h or exposed to pemphigus serum for 72 h and then cultured in normal serum for 2 wk. Oral epithelial cells from the pemphigus patient continued to display acantholysis for the 3-wk duration of the culture. Normal oral epithelial cells showed acantholysis within 30 min exposure to pemphigus serum and the acantholysis persisted in the cultures, pulse exposed to pemphigus serum, for the duration of the explant culture. These results support the notion that pemphigus serum is directly responsible for acantholysis which may then be persistent.

Keratinocyte culture.

The in vitro growth of keratinocytes has proved to be an important tool in the study of the normal biology and disease processes involving the skin, e.g., the influence of extrinsic regulators of growth and differentiation, effects of pharmacological agents, dermo-epidermal interactions, tissue antigenicity, and models of carcinogenesis.

T suppressor efferent circuit which affects contact sensitivity to picryl chloride: the late-acting, second nonspecific T suppressor factor bears I-A determinants which are responsible for the I-A genetic restriction in its interaction with its target cell.

The T suppressor efferent circuit in the picryl (TNP) system, which inhibits the passive transfer of contact sensitivity, involves at least two antigen-nonspecific factors. The second nonspecific T suppressor factor (ns-2) bears I-A determinants of both the alpha and the beta chain as shown by affinity chromatography on immobilized anti-I-A monoclonal antibodies. Sequential absorption shows that the determinants of the alpha and beta chain occur on the same molecular complex. No absorption was obtained with anti-I-E antibody. There are two genetic restrictions associated with ns-2--the first is in its release from the second T suppressor efferent cell (on exposure to antigen) and the second is in its inhibitory interaction with its target cell. Both are MHC restricted and matching in I-A (but not I-E, or I-J) is sufficient. The question was asked whether the I-A of the ns-2 was directly responsible for the I-A genetic restriction in its action. F1 TsF was made in (H-2k X H-2b)F1 mice by injecting picrylated parental cells intravenously and triggering the release of ns-2 with the corresponding picrylated parental cells. Both I-Ak- and I-Ab-positive ns-2 were produced and were separated by affinity chromatography on immobilized anti-I-A monoclonal antibody. The I-A phenotype of these separated ns-2 of F1 origin determines the genetic restriction in their action; i.e., I-Ak+ ns-2 only inhibits passive transfer by H-2k cells and I-Ab+ ns-2 only acts on H-2b cells. In contrast, the I-A haplotype of the picrylated cell used to induce the Ts cell which makes ns-2 is unimportant. It was concluded that the I-A on the ns-2, and not a possible recognition site for I-A, serves as a restriction element. This finding suggests that ns-2 may act directly on the I-A-restricted T cell which mediates contact sensitivity.