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.

Potentially difficult smears of women with squamous cell carcinoma pose fewer problems when PAPNET is used for primary screening.

The diagnosis of squamous cell carcinoma (SCC) on a cervical smear is often far from easy. This study reports the analysis of 40 true-positive SCC smears detected in primary PAPNET screening and eight false-negative (FN) conventionally screened smears. All FN cases contained sparse abnormal material (< 10% of the slide). In these potentially difficult cases the diagnosis on the PAPNET images was not hard. Statistical analysis of the quantitative data indicated that the PAPNET images of the FN cases and the true-positive cases differed in some aspects. PAPNET highlighted the importance of background information (old blood, fibrin and necrosis). In addition, all FN smears contained cancer cells in the PAPNET images, allowing a correct diagnosis.

Analysis of lifestyle data and cytologic findings in a pilot cervical screening project in rural Vietnam.

OBJECTIVE: To evaluate the possibilities of a cervical cytology screening program and the importance of lifestyle parameters in women living in rural Vietnam. STUDY DESIGN: Screening took place in Quangninh province in the north of Vietnam. From 700 women screened for the first time, smears were prepared and read by a cytotechnologist. The number of children and abortions (some women having had 10 abortions) allowed us to study the relationship of these life-style parameters with disturbed vaginal ecology (defined as the presence of Candida, trichomonads or cocci in the smear). RESULTS: The smears from farmers showed the greatest ecologic disturbances in vaginal flora. Water supply turned out to be important for coccoid overgrowth. Scores for both fungal infection and coccoid overgrowth were related to profession. Candidosis proved to be very prominent in cytologically negative smears: the organisms were found (often in abundance) in 95 of 661 negative smears but were absent from cytologically positive smears. CONCLUSION: Since only two cases of CIN 3 were found and no cases of invasive carcinoma, cervical screening for cancer ought not to be the first priority for rural Vietnam.

Cytotoxic T lymphocytes and antibodies after orthotropic penetrating keratoplasty in rats treated with dichloromethylene diphosphonate encapsulated liposomes.

PURPOSE: To investigate the immunological basis for the prolonged corneal allograft survival after subconjunctival injections of liposomes filled with dichloromethylene diphosphonate (Cl2MDP-LIP). METHODS: F344 rats received orthotropic DA corneal grafts. One group of rats was treated with subconjunctival injections of Cl2MDP-LIP on days 0, 2, 4, 6 and 8 postoperatively, the control groups received no treatment. Nineteen or 42 days postoperatively cytotoxic T lymphocyte (CTL) activity was measured in the lymph nodes draining the grafted and the contralateral eyes, in the spleen and the mesenteric lymph nodes. Sera taken at the same time points were tested for presence of lytic alloantibodies. RESULTS: On day 19 CTL activity in submandibular lymph nodes draining the grafted eyes was similar in the 2 groups. In the mesenteric lymph nodes high CTL activity was found in the untreated rats and low in the Cl2MDP-LIP rats. The spleen showed high CTL activity in the untreated group but no activity in the liposome group. Forty two days postoperatively a decline in CTL activity was seen in both groups. Complement dependent anti-donor antibodies were absent in the Cl2MDP-LIP group at both time points whereas antibodies were present on days 19 and 42 in the untreated group. CONCLUSIONS: Repeated subconjunctival injection of Cl2MDP-LIP restricts the induction of cellular cytotoxicity against donor antigens to the regional lymph nodes and suppresses cytotoxic antibody formation.

Influx of immunoglobulins from the vascular compartment into a grafted cornea.

PURPOSE: To determine the effect of a fresh corneal wound or a healed corneal scar on the immunodiffusion of immunoglobulins into the cornea. METHODS: F344 rats were immunized with human serum albumin (HSA) 1 week before an autologous rotational keratoplasty of the right cornea or 1 year after an autograft was performed. One group of rats also was treated with gentamicin-dexamethasone ointment in the grafted eye for 1 week after transplantation to reduce the postsurgical inflammatory signs. A serum sample was drawn every week and booster injections with HSA were given after 2 and 3 weeks. At various times after immunization, groups of rats were killed, blood and aqueous humor samples were taken, and the corneas of both eyes were removed. The corneas were divided into the graft or a 3-mm central button and the peripheral rim and weighed. The anti-HSA titer was determined in serum, aqueous humor, and both parts of the corneas. RESULTS: Up to 5 weeks after transplantation, the grafted cornea contained more anti-HSA immunoglobulins than did the control eye. One year postgrafting, no difference was seen. In the first weeks after keratoplasty, influx of anti-HSA from the peripheral into the central cornea was, however, neither obstructed nor enhanced. CONCLUSIONS: Surgical trauma in itself causes increased influx of anti-HSA immunoglobulins into the cornea. Within the cornea, a wound or a scar does not appear to be a barrier for centripetal immunoglobulin diffusion.

Prevention of corneal allograft rejection in rats treated with subconjunctival injections of liposomes containing dichloromethylene diphosphonate.

PURPOSE: The drug dichloromethylene diphosphonate (CL2MDP) encapsulated in liposomes depletes macrophages but not other immunocompetent cells. The authors investigated whether subconjunctival injection of CL2MDP containing liposomes (CL2MDP-LIP) could prolong survival of corneal allografts in rats. METHODS: Male Fisher rats received orthotopic corneal grafts of Dark Agouti origin. Rats were treated postoperatively with subconjunctival injections of 0.1 ml CL2MDP-LIP at the time of transplantation and on days 2, 4, 6, and 8 after transplantation. Control groups received either liposomes containing phosphate-buffered saline subconjunctivally at the same time points or no additional treatment. Corneal grafts were evaluated every other day and were scored for neovascularization, opacity, and edema. Immunohistologic evaluation was performed 12 and 19 days after surgery. RESULTS: Corneal grafts in both control groups were rejected within 17 days. In the Cl2MDP-LIP treated rats, grafts were not rejected during the maximum follow-up of 100 days. Cellular infiltration in these grafts was clearly reduced. There was also a strong reduction in neovascularization of the cornea. CONCLUSIONS: Rejection of orthotopic allogeneic corneal grafts could be prevented by repeated subconjunctival injection of Cl2MDP-LIP.

New material for low-cost intraocular lenses.

A UV-hardening lacquer material based on polyurethane, used in Philips compact disc lens systems, was tested as suitable material for low-cost intraocular lenses (IOLs). A slightly changed composition (code number I-0.5A) came out as the best and was subsequently tested, with reference to poly(methyl methacrylate) (PMMA), as a 'lobster-claw' lens in Hollander rabbit eyes for a period of 24 wk. No carcinogenic effects were found, while only minor vascularization of the cornea at the incision area occurred. The in vitro biocompatibility tests of the material were positive. Scanning electron microscopy images of the IOLs at 2, 6, 12 and 24 wk postoperative showed portions of an acellular membrane, multinuclear giant cells which hyalinize and stay as pigmented plaques, microphages and fibroblasts with fibrillar strands, long pseudopodia, and broad microplicea. The chosen UV lacquer showed a foreign body reaction and properties comparable with PMMA, and is suitable for manufacturing low-cost IOLs.

Cell-ingrowth in a silicone plombe. Interactions between biomaterial and scleral tissue after 8 years in situ: a SEM and TEM investigation.

A male patients (42 years) who had been treated for retinal detachment by the implantation of a silicone plombe into the sclera, returned to the clinic 8 years after implantation because of inflammation of the tissue and partial protrusion from the sclera. After removal of the plombe this was processed for TEM and SEM and examination of the plombe material after 8 years in situ could be carried out in order to get information about the cell-biomaterial interface. A large proportion of the silicone pores was filled with cellular material, including macrophages, giant cells and erythrocytes thus indicating a foreign body granuloma. The external surface of the pores showed a granular osmiophilic dense amorphous layer including extracellular debris. Engulfing of silicone particles by macrophages and the evidence of long-term tissue response suggest partial biodegradation of the silicone and certainly not complete inertness as was formerly claimed.

A combined scanning and transmission electronmicroscopic investigation of human (secondary) cataract material.

Cataract lenses from patients of advanced age were processed for SEM by standard pre-fixation followed by treatment by the Tannin-Arginine-Osmium-tetroxide (TAO) method and critical point drying, and for TEM by standard pre-fixation followed by vibratomation, standard post-fixation, ultramicrotome sectioning and staining with uranyl acetate/lead citrate. Secondary cataract material was brought onto a Millipore filter, fixed by standard methods, dried in air and sputter-coated with Au. Both SEM and TEM images revealed degeneration processes in lensfibre material, such as swelling of the lensfibre, protrusion of the cytoplasm, fibrillation of the cell membrane, loss of the nucleus, spherical bodies of various sizes between 0.5-1.5 microns, sometimes surrounded by a (double) membrane with different contrast but without cellular evidence, and small and large vacuoles partly filled with granular material both in and at the periphery of the lensfibre-body. The secondary cataract material on the Millipore filter revealed erythrocytes and more or less spherical bodies with high contrast, measuring between 0.5-1.5 microns, often referred to as Elschnig's pearls, besides non-definable organic material. The SEM and TEM micrographs of the cataract lens material strongly suggest that the spherical bodies with sizes of approximately 0.5-1.5 micrometer and high contrast without cellular evidence, are similar to the more or less spherical bodies found in the secondary cataract material on the filter, referred to as Elschnig's pearls.

Morphological aspects of human lens capsules. A comparative LM, SEM and TEM examination.

Lens capsules of patients of advanced age, obtained after extracapsular cataract surgery, were carefully prepared for a combined LM, TEM and SEM investigation, after preliminary washing and mounting onto a holder in a buffer solution. After pre-fixation with GA, samples were postfixed for LM/TEM and OsO4/K4Fe)CN)6 and stained with toluidine-blue/basic fuchsin for LM and with uranyl acetate/lead citrate for TEM; for SEM the GA-pre-fixed samples were post-fixed by the Tannin Arginine-OsO4 non-coating technique. At LM-level discrimination between healthy and degenerating cells was possible after toluidine staining. At SEM-level protrusion of the cell nucleus and fibrillation and blebbing of the cell membrane as the result of capsular degeneration could be observed with the TAO-method. At TEM-level protrusion of the cell nucleus, degeneration of the cytoplasm, ballooning of the mitochondria, the presence of microfilaments and the occurrence of vacuoles were visible as the result of capsular degeneration on cataract formation.

Refractive surgery for high myopia. The Worst-Fechner biconcave iris claw lens.

Surgical correction of high myopia is indicated in a limited number of cases. Techniques include (epi)keratophakia, keratomileusis and implantation of a negative-power IOL. The iris claw lens, fixed to the midperiphery of the iris, has proved its safety in aphakic eyes. The model has been adapted by giving it a vault in order to make it suitable for implantation in high myopic phakic eyes. The implantation technique and short-term results of patients operated by Worst are presented.

Reaction of the rabbit corneal endothelium to nylon sutures. A SEM study.

Nylon and stainless steel sutures separately placed deeply into rabbit corneas by splitting the stroma for a few millimeters, without closing sutures, remained in the cornea for two, four and six weeks respectively. In contrast to the stainless steel sutures an extensive tissue reaction could be observed clinically around the nylon sutures within a few days and was still present after 4-6 weeks. On the endothelial side, covering of the nylon with fibroblast cells took place very slowly, this in contrast with stainless steel. Irritation of the tissue and an oedematous appearance of the endothelium around the nylon suture was the result. At first a kind of collagen network, often mixed with inflammatory cells, was deposited on the nylon material before fibroblast cells could grow in. Covering of the stainless steel started quite soon after implantation, without preliminary deposition of collagen material. The reason for this phenomenon must be sought in the high free surface energy of the stainless steel, which attracts cells, in contrast to nylon which has a very low free surface energy.

The Gonioseton, a surgical treatment for chronic glaucoma.

A new and simple technique for glaucoma filtering surgery is presented. It is based on the introduction of a stainless steel microspiral or gonioseton into a goniopuncture by way of an ab interno procedure, minimising surgical trauma to the filtration area. Preliminary results are presented.

Secondary cataract material collected with a glass cannula. A SEM study.

Secondary cataract material from three patients, collected with a glass cannula approx. 18, 24 and 30 months respectively after operation, was prepared for SEM examination. For the soft samples this was done by filtration through a millipore filter followed by fixation and drying. The more solid material was suspended in a fixation solution, followed by centrifuging, suspension in 70% ethanol and drying on a specimen-holder. The short residence samples (18 months) showed mainly erythrocytes, some (inflammatory) cells and degenerated lens-fibre material. Most of the more solid material, which was collected more than 20 months after operation, showed fragments of (regenerated) capsule epithelium and pieces of solid lens-fibre material with fragments of capsule epithelium attached.

Progesterone in obese patients with respiratory failure.

Two obese patients with acute respiratory failure are described. Both required mechanical ventilation to reverse the respiratory acidosis. On recovery, the major abnormality was failure of ventilatory response to inhaled carbon dioxide. After oral progesterone (100 mg/day and 20 mg/day, respectively) the slope of the carbon dioxide response line increased from 0,02 to 1,14 I min-1 mmHg-1 and from 0,04 to 1,14 I min-1 mmHg-1, respectively. This was associated with sustained clinical remission in 1 patient.