[Enterobius gregorii Hugot, 1983 recovered from school children in Kangwon-Do, Korea]. Hugot (1983), Hugot and Tourte-Schaefer (1985) reported that nematodes of the genus Enterobius causing human enterobiasis might be classified into two different species on the basis of the size and shape of spicule in male worms. E. gregorii Hugot, 1983 has significantly shorter spicules in males than those in E. vermicularis. A total of 194 male Enterobius were collected from 17 schoolchildren by anthelmintics at Wonju, Kangwon-do. The sizes of spicules and morphologies were comparatively observed. Two different types of spicules in male Enterobius were also observed in Korea. Out of 112 male worms observed, the percentages of E. gregorii (short-spicule type) and E. vermicularis (long-spicule type) were 23.2% and 76.8%, respectively.


Egg production of Clonorchis sinensis in different strains of inbred mice. In order to compare the intraspecific variation in host-parasite relationship of Clonorchis sinensis, six strains of inbred mice, ICR, DDY, GPC, BALB/c, nude and DS, were infected orally with 20 metacercariae of C. sinensis. The biologic incubation period of C. sinensis was the shortest in DDY mice, 21.2 days in average, followed by GPC 21.4, BALB/c and DS 23.2, ICR and nude 23.4 days, respectively. The fertile period of the fluke was also the longest in the DDY strain, 164 days on average, followed by GPC 132, BALB/c 97, nude 37, DS 32 and ICR 28 days. The egg-laying capacity of the fluke in DDY and GPC was relatively high and stable compared with the other four strains of mice. It was found that there are intraspecific variations in biologic incubation period, fertile period, and fecundity of C. sinensis. The DDY mouse is likely to be the most suitable experimental animal among the six strains of the mice tested.


Platelet kinetics and other hematological profiles in experimental Plasmodium falciparum infection: a comparative study between Saimiri and Aotus monkeys. Levels of platelets and other hematological values were monitored in 21 Saimiri and 12 Aotus monkeys over a period of three weeks post-infection with monkey-adapted Indochina CDC-1 strain of Plasmodium falciparum. In both Saimiri sciureus boliviensis and Aotus nancymai karyotype-1 monkeys the severest thrombocytopenia was observed at 14 days post-infection coinciding with peak parasitemia, neutropenia, lymphocytosis, and anemia associated with severe hemoglobinemia and elevated fibrinogen degeneration products(FDP's). MCH and MCV profiles in Aotus monkeys decreased with ascending parasitemia. In contrast, these parameters in Saimiri were characterized by a significant compensatory increase correlating with parasitemia. In general, thrombocytopenia was one of the earliest clinical manifestations of the infection with the platelets returning to normal levels shortly after peak parasitemia at 14 days. Platelet kinetics had a strong correlation with hematologic and parasitologic values in the Aotus model. No consistent associations were observed between platelet kinetics and other parameters in the Saimiri model. These data indicate that the Aotus model for malaria is more predictable than the Saimiri. Further, platelet turnover rates and recovery provide a useful prognostic parameter during malaria infection. The results are discussed in relation to the value of the two species of monkeys as models for the pathogenesis of human malaria.


Karyotypes of Pneumocystis carinii from Korean rats. Molecular karyotyping was applied to Pneumocystis carinii(Pc) from two strains of experimental rats, Sprague Dawley(SD) and Fisher(F), in Korea. Field inversion gel electrophoresis and contour clamped homogeneous electric field electrophoresis resolved 15 chromosomal bands from the Pc. The size of the bands was estimated 270kb to 684kb from SD rats, and 273kb to 713 kb from F rats. The bands of 283 kb from SD rats and of 273 kb from F rats stained more brightly suggesting duplicated bands. Total number of chromosomes was at least 16, and total genomic size was estimated 7 x 10(6) bp. All of the bands from F rats hybridized to the probe of repeated DNA sequences of Pc and the band of 448 kb size was proved to contain rDNA sequences, but Pc. chromosome bands from SD rats showed no reactions to the probes. The 2 different karyotypes of P. carinii from 2 strains of rats were maintained consistently for 2 years.


Isolation and partial characterization of cysteine proteinase from sparganum. A proteolytic enzyme was purified from the tissue extract of spargana (plerocercoids of Spirometra erinacei) by DEAE-Trisacryl M ion exchange chromatography and thiopropyl-sepharose affinity chromatography resulted in a 21-fold purification. The proteinase activity was assayed with a synthetic fluorescent substrate, carbobenzoxy-phenylalanyl-7-amino-4-trifluoromethyl-coumarin. SDS-polyacrylamide gel electrophoresis of the purified materials revealed a single 28,000 dalton band. Inhibitor profiles of the band indicated that it belonged to cysteine endopeptidases. It exhibited identical pH curves with optimum at pH 5.5, and 50% activity from pH 4.7 to 8. It could completely degrade collagen chains to three identical products. It also showed some activity on hemoglobin. Furthermore, the band on immunoblots was reactive to the sera of sparganosis patients. These results suggest that the proteolytic enzyme belongs to cysteine proteinase which plays a role in the tissue penetration. Also it may be used as the antigen for diagnosis of active sparganosis.


Immunogenicity and protective efficacy of solubilized merozoite-enriched Theileria sergenti immunogens. II: Protection against natural exposure under field conditions. A Theileria sergenti soluble merozoite preparation containing the 29, 34, 35 and 105 KD as the immunodominant polypeptides, was evaluated for efficacy, safety and protectivity in Holstein calves against virulent field tick challenge. The soluble antigens (100 mg/dose) were fortified with either complete or incomplete Freund's adjuvant. Twenty naive calves, aged one month, were subcutaneously inoculated with the preparation and a booster dose was administered 4 weeks later. Twenty additional calves served as controls. Five weeks after the booster dose, vaccinates and uninoculated controls were moved to a pasture, a heavily tick infested area in Cheju-do, Korea. The vaccinates showed negligible change in hematocrit and total RBC count whereas control animals showed significant (p < 0.05) hematological changes and associated anemia. Only 30% of vaccinates required chemotherapy after the experiment was terminated. All control animals required chemotherapy and 25% received blood transfusion. The highest percent parasitized erythrocytes in vaccinated cattle was 0.4% as compared with 3.6% among controls during the month of July. A significant difference (p < 0.05) was observed in the rate of body weight increase. Significant differences were also noted in serum albumin, aspartate aminotransferase, total protein and bilirubin. Significantly more vaccinated cattle maintained normal ranges of hematological and biochemical values as compared with the control group. It is suggested that soluble merozoite T. sergenti antigens may be potential vaccine candidates for developing a genetic vaccine in Korea.


Immunoelectrophoretic analysis of major component proteins in cystic fluid of Taenia solium metacestodes. When cystic fluid of Taenia solium metacestodes (CF) was filtrated through Sephacryl S-300 Superfine, major proteins were in fractions III and IV. Major protein in fraction III was Band C protein of 150 kDa and that in fraction IV was Band N protein (Choi et al., 1990). When CF was electrophoresed in 0.9% agarose gel and reacted with anti-CF rabbit serum (RACF), two main bands, a long outer and a short inner band, were precipitated, together with 8 minor bands. RACF reacted with fraction III forming the long outer band whereas RACF formed the short inner band with fraction IV in immunoelectrophoresis (IEP). The long outer precipitin band of CF fraction III was similar to antigen B in hydatid fluid (HF) of Oriol et al. (1971), while the short inner band of CF fraction IV was similar to HF antigen 5 of Capron et al. (1967). When HF was reacted with RACF, the short inner band was immunoprecipitated without forming the long outer band. Common antigenicity between CF and HF seemed to exist in fraction IV rather than in fraction III of CF. Patient sera of neurocysticercosis reacted more frequently with fraction III than with fraction IV.


Component proteins and protease activities in excretory-secretory product of sparganum. Spirometra mansoni plerocercoid (sparganum) was incubated in saline at 4 degrees C or 37 degrees C up to 100 hours. Protein contents in the excretory-secretory product (ESP) were rather constant (mean 7.7 mg of protein/gram of sparganum) in the preparations. Reducing SDS-PAGE of ESP showed similar protein subunit compositions with those in crude extract. Antigenic 36 and 31 kDa proteins were major bands in ESP. ESP exhibited specific activities of protease (2.9-5.3 units/mg) at pH 6.0 and pH 7.5. Presence of protease activity in ESP may be a supporting evidence that hitherto known cysteine protease of sparganum is possibly secreted.


[A case of strongyloidiasis accompanied by duodenal ulcer]. A 58-year-old chronic alcoholism patient, who complained of epigastric discomfort, nausea, and frequent loose stool was diagnosed as strongyloidiasis accompanied by duodenal ulcer. The symptoms first appeared two years ago and aggravated during the recent 3 months, and he lost 4 kg of his body weight. Stool examination revealed rhabditoid nematode larvae, which were confirmed as those of Strongyloides stercoralis after cultivation of them to filariform larvae. At duodenoscopy, duodenal ulcer was found. The patient was treated with albendazole (200 mg, bid, for 14 days) for strongyloidiasis and with colloidal bismuth sulfate (240 mg, bid, for 6 weeks) for duodenal ulcer. After the medication, the symptoms of loose stool and epigastric discomfort were much improved and he was discharged with no clinical problems. This is an interesting case which suggests that S. stercoralis infection could be related with ulceration of the duodenal mucosa.


The red eye of renal failure: a crystal induced inflammation? Of 57 patients with chronic renal failure and calcification of the anterior membranes of the eye eight developed inflammatory reactions, clinically indistinguishable from pingueculitis. In three patients an acute inflammatory reaction of the episcleral tissue and the conjunctiva over it developed which we considered to represent the red eye of renal failure. In these patients massive shedding of calcium phosphate salts was clinically evident. Histopathologically in all these patients calcific deposits were observed. Although sporadically polymorphonuclear leucocytes were present in the inflamed tissues we could not demonstrate crystal phagocytosis. We believe that in the red eyes in renal failure and, more specifically, in the red eyes of renal failure a crystal induced inflammatory mechanism is not operative.


Herpetic keratitis in Tanzania: association with malaria. Of 103 patients who presented to a rural clinic in Africa with corneal ulceration, 62 (60.2%) had corneal ulcers which on clinical diagnosis were attributable to herpes simplex virus. There was a strong association between herpetic ulceration and a history of recent malaria; 37 of 62 (59.7%) herpetic ulcers occurred in the 3 month period from April to June which corresponds to the end of the peak season for malaria compared with 14 of 41 (34%) of the non-herpetic ulcers. Fifty per cent of herpetic ulcers had a geographic morphology, 27.4% were dendritic, and 22.6% presumed herpetic ulcers were stromal: 38.7% of herpetic ulcers occurred in children under 5 years of age. Bilateral herpetic ulcers occurred in 16.1% of patients but were more common in children under 2 years of age. Geographic and stromal ulcers tended to heal more slowly than dendritic ulcers (mean time to healing 12.6, 12.2, and 6.6 days respectively), and were more likely to result in severe corneal scarring (45%, 29%, and 0% respectively). Herpes simplex keratitis is a major cause of corneal scarring in Africa. It is often seen in children, may be bilateral, commonly is geographic in morphology, and has a strong association with malaria infection. Because it is not easily preventable, more effort must be made to ensure early diagnosis and prompt, effective treatment in order to prevent severe scarring and visual loss.


Radial keratotomy: a review of 300 cases. Three hundred consecutive cases of radial keratotomy performed between 1985 and 1990 were reviewed. There were no sight threatening complications of surgery and no patient lost one or more lines of corrected Snellen acuity. Overall 78.7% saw 6/12 or better unaided postoperatively and 51.7% saw 6/6 or better. Refraction showed 61.3% to be within 1 dioptre of emmetropia and 86.7% were within 2 dioptres. Further analysis demonstrated that results of unaided acuity and proximity to emmetropia were much better for low (< -2.87 D) and moderate (-3.0 to -5.87 D) than for high (> -6.0 D) myopes.


Life expectancy in keratoconus.

Life expectancy in keratoconus. It is observed that few patients over the age of 60 regularly attend the keratoconus clinic at Moorfields Eye Hospital. The hypothesis that patients with keratoconus have a shorter life expectancy owing to underlying connective tissue related disease was tested. From patient records a sample of 337 keratoconus patients aged at least 40 years by 1991 were identified, of which 279 were living, 13 were deceased, and 45 were untraceable. The mortality rate for keratoconus patients was compared with that of the general population using actuarial English life tables. Results show no significant difference between the general population mortality rate and that of the keratoconus sample even with adjustment for social class. Possible explanations for the non-attendance of older patients are discussed.


Foveal photopigment kinetics--abnormality: an early sign in myotonic dystrophy? Twelve subjects with minimal expression of the myotonic dystrophy (MyD) gene were investigated by retinal densitometry, a technique which has been used to study the properties of photopigments in the living eye and to detect photoreceptor abnormalities. Other investigations included slit-lamp examination, funduscopy, raleigh matches with the anomaloscope, tonometry, and neurological examination, including electroretinography (ERG) and pattern visual evoked potentials recording. Foveal densitometry demonstrated reduced values of the macular photopigment density difference with normal photopigment kinetics in early phases of the disease, even in asymptomatic individuals. The densitometric values correlated with decreased amplitudes of the photopic ERG a-wave. These findings may be explained by loss or dysfunction of the outer segments of foveal receptors. It is yet unknown whether or not these changes are secondary to other observed neuroretinal abnormalities in MyD. The most likely explanation might be an abnormality of the Na, Ca:K exchanger at the level of the outer segments of the photoreceptors whether or not in combination with a dysfunction of voltage generation systems, involving both photoreceptors and retinal pigment epithelium.


Intravenous sedation for ocular surgery under local anaesthesia. Anterior segment ophthalmic surgery is commonly performed under local anaesthesia. In order to improve patient comfort, a variety of sedation techniques has been employed in the past. The object of this study was, firstly, to determine whether continuous intravenous sedation during surgery offered any advantages in patients premedicated with temazepam and metoclopramide, and, secondly, to compare midazolam to propofol for this purpose. Forty nine patients were randomly allocated to receive no intravenous sedation (n = 15), continuous propofol infusion (n = 17), or continuous intravenous midazolam infusion (n = 17) after peribulbar anaesthesia. Each technique provided cardiovascular and respiratory stability and allowed early recovery with minimal postoperative sequelae. Unexpected ocular field movement occurred more commonly in the patients receiving intravenous sedation, although statistical significance was not shown (p = 0.06). Significantly more patients in the intravenous sedation groups reported amnesia (p = 0.03). Patient acceptability was good irrespective of the technique used. This study suggests that continuous sedation using propofol or midazolam is not beneficial and should be avoided in ophthalmic patients who have received a simple premedication.


Type IV collagen and laminin in Bruch's membrane and basal linear deposit in the human macula. Tissue obtained from the macula in 10 human eyes (53-77 years) was used for an investigation into the extracellular matrices of the retinal pigment epithelium (RPE), Bruch's membrane, and the choriocapillaris. The ultrastructural distribution of type IV collagen and laminin was documented using immunogold labelling. Labelling for type IV collagen was strongly positive in all the specimens in the basement membranes of the choriocapillaris but not that of the RPE where labelling was either weak or absent. Laminin was localised to deposits of granular material in Bruch's membrane but was absent from the basement membrane of the RPE and the choriocapillaris. Basal linear deposit, observed in three cases, demonstrated labelling for laminin but not for type IV collagen. The series was too small for correlation of these morphological changes with age.


Induced chorioretinal venous anastomosis in experimental retinal branch vein occlusion. Iatrogenic retinal vein to choroidal vein anastomoses were created using laser photocoagulation in six of seven dog eyes in which a partial branch retinal vein occlusion had previously been created photochemically. A similar attempt to create an anastomosis was made in six control eyes in which no branch vein occlusion was present. In the eyes in which a branch retinal vein had been created, a venous chorioretinal anastomosis appeared to be present by 3 to 6 weeks. In three control eyes similar venous anastomosis was created; however this took 6 to 8 weeks to develop and was of much smaller calibre than the one that developed in the presence of a partial branch retinal vein occlusion. No adverse complications were noted in the period of the study (3 months). This study demonstrates that chorioretinal venous anastomoses can be created and may be of use in the treatment of partial retinal vein occlusions that show signs of progression.


AgNOR counts in conjunctival malignant melanoma lack prognostic value. Using a silver staining technique, argyrophilic nucleolar organiser region-associated proteins (AgNORs) have been studied in routinely processed paraffin sections of 46 invasive malignant melanomas (MM) of the conjunctiva. The aim of this retrospective study was to assess the value of the AgNOR method as a prognostic indicator for this neoplasm. The 46 cases were divided into two groups: (A) 14 cases of MM that metastasised and caused death of the patient within 5 years of (histological) diagnosis, and (B) 32 cases of MM that did not metastasise and in which patients survived beyond 5 years. The mean of the AgNOR counts per nucleus was 7.03 (95% CI: 5.81-8.24) in group A, and 7.15 (95% CI: 6.53-7.77) in group B. A comparison using a multifactor analysis of variance (ANOVA) model, which corrected for possible confounding effect of tumour thickness, site, and cell type showed no significant difference in AgNOR counts between groups A and B (p = 0.8). Analysis by the Cox proportional hazards regression model showed that survival was not influenced significantly by the mean AgNOR number (hazard ratio: 0.92). Whereas the AgNOR technique may be used to distinguish benign from malignant melanocytic lesions of the conjunctiva, we conclude it has no value in predicting the outcome for patients with conjunctival MM.


Laser trabeculoplasty.

Laser trabeculoplasty. Using the now well established technique ALT is a safe and effective means of lowering IOP in eyes with POAG, pigmentary glaucoma, and exfoliative glaucoma though the effect diminishes with time. Pressure reduction with ALT is seldom greater than 30% and eyes with an IOP of more than 28 mm Hg are unlikely to be controlled for any length of time without additional treatment. ALT is probably most suitable either for older patients whose glaucoma is not well controlled on medical treatment, or for patients with newly diagnosed open angle glaucoma, but without advanced field loss, as a primary treatment or in combination with a non-miotic topical medication. In many it will defer surgery, in some indefinitely.


Bilateral multiple cavernous haemangiomas of the orbit. Orbital cavernous haemangioma, a common orbital tumour, is usually single and unilateral. We report the first case of histologically confirmed bilateral multiple cavernous haemangiomas of the orbit.


Vanishing retinal arterial aneurysms: a case report. An 18-year-old woman with retinal vasculitis developed multiple retinal arterial aneurysms over a period of 3 years. Subsequently she developed macular oedema, peripheral neovascularisation, and vitreous haemorrhage and was treated with systemic steroids, laser photocoagulation, and vitreoretinal surgery. No systemic cause for vasculitis was found. Serial fluorescein angiography demonstrated gradual resolution of the arterial aneurysms over the subsequent 7 years.


The electro-oculogram in central retinal vein occlusion. As part of a prospective masked study, the electro-oculogram (EOG) was recorded from 28 patients within 48 days of developing central retinal vein occlusion (CRVO). The EOG light peak/dark trough ratio (Lp/Dt) x 100 was significantly lower in the affected than in the unaffected eyes of patients (p < 0.001), and abnormally low in absolute terms in 20 patients (71%). All unaffected fellow eyes had a normal EOG ratio. The mean Lp amplitude of affected eyes was significantly smaller than that of unaffected eyes (p < 0.001), whereas the differences in mean Dt amplitudes between affected and unaffected eyes were not statistically significant. The Lp amplitude in the affected eye was 48% or less of that in the unaffected eye in the eight patients (29%) who developed rubeosis iridis during the 9 month follow-up, and in six others. No patient whose Lp amplitude in the affected eye was greater than 48% of that in the unaffected eye, developed rubeosis. It is concluded that the Lp amplitude is abnormal in patients with acute CRVO. The degree of this abnormality bears a relation to the development of rubeosis, which might prove a useful indicator of whether to institute or withhold panretinal photocoagulation.


Controlled study of ocular morbidity in school children born preterm. Two hundred children who were of very low birth weight (VLBW) (1500 g or less) and 193 controls who were of normal birth weight (NBW) were examined at approximately 9 years of age. Binocular visual acuity of 6/6 or better was noted in 178 (89.5%) VLBW children and 189 (97.9%) NBW children. Visual morbidity was significantly higher among VLBW children. Strabismus was present in 38 (19%), cicatricial retinopathy of prematurity in 13 (6.5%), and optic atrophy in six (3%) children in this group. Children who were VLBW were also more myopic than the NBW controls. Optic atrophy was frequently associated with cerebral dysfunction. Regular assessments to identify ocular abnormalities in children who were VLBW are recommended.


Vitreon, a short-term vitreoretinal tamponade. This investigation of the liquid perfluorocarbon, perfluorophenanthrene (Vitreon), establishes its safety and efficacy as a short-term vitreoretinal tamponade. We utilised Vitreon as an intraoperative tool and postoperative vitreoretinal tamponade in 16 patients. Proliferative vitreoretinopathy (PVR) (six), giant retinal tear (four), rhegmatogenous retinal detachment (three), retinal detachment with keratoprosthesis (two), and submacular and vitreous haemorrhage (one) were successfully repaired. Vitreon was left in the eye and removed 5 days to 4 weeks postoperatively. Complications encountered included proliferative PVR (five), limited peripheral retinal detachment (three), macular pucker (two) cataract (three), hypotony (two), excessive fibrin reaction (one), and elevated intraocular pressure (one). At the latest evaluation, all retinas are attached with a follow-up of 1.25 to 12 months (mean 6.8 months).


Ocular circulatory changes following scleral buckling procedures. The effect of segmental scleral buckling (SB) on ocular circulation was evaluated by measurements of the ocular pulse amplitude (PA) and the ophthalmic artery pressure (OAP). In this study the OAP was defined as the intraocular pressure (IOP) at which the PA disappeared during increasing IOP. Twenty four patients with unilateral rhegmatogenous retinal detachment who underwent SB of varying extent were studied; the unoperated fellow eyes served as controls. Both the OAP and the ophthalmic perfusion pressures (OAP minus IOP) decreased significantly (p < 0.01) as the area of SB treatment increased. Follow-up measurements showed that the OAP was relatively lower up to 3 months postoperatively than after 3 months, and then remained stable. Our results indicated that SB affects the ocular pulse measurements and may decrease ocular blood flow because of decreased ophthalmic perfusion pressure, probably due to increased choroidal vascular resistance, and that the greater the extent of the SB treatment, the greater the possibility of decreased ocular blood flow.


Cautery for lower lid entropion.

Cautery for lower lid entropion. A prospective study was undertaken to evaluate a simple cautery technique for the correction of involutional lower lid entropion in 50 patients. After a 12 month follow-up period all patients were free of entropion. Only one patient needed to have the procedure repeated because of recurrence. The technique was found to be simple, effective, safe, and required very little time and skill.


Prevention of acute postoperative pressure rises in glaucoma patients undergoing cataract extraction with posterior chamber lens implant. Acute elevations in intraocular pressure (IOP) commonly follow extracapsular cataract extraction and lens implant in glaucoma patients. Thirty six patients with glaucoma undergoing cataract extraction and posterior chamber lens implantation received one of three treatments. Group 1: 500 mg of Diamox Sustets (acetazolamide) 1 hour preoperatively (10 patients); Group 2: peroperative intracameral Miochol (acetylcholine) (11 patients); Group 3: the above treatments combined (15 patients). IOPs were measured at 3, 6, 9, and 24 hours postoperatively. The average of the maximum pressure rises above the preoperative level over the 24 hour period was greatest for the group receiving acetazolamide only at 8.9 mm Hg; for the acetylcholine group the average maximum rise was 6.3 mm Hg; while the combined treatment group showed a decrease of 0.7 mm Hg. IOP rises of > 6 mm Hg were seen in 7% of patients (one of 15) in the combined treatment group, 45% (five of 11) of the acetylcholine group, and 70% (seven of 10) of the acetazolamide group. IOP rises of > 10 mm Hg were seen in 7% of the combined treatment group, in 18% of the acetylcholine only group, and in 50% of the acetazolamide only group. A pressure rise > 20 mm Hg was seen in one patient receiving acetazolamide only and one patient receiving acetylcholine only. The difference between the acetylcholine group and the combined group for rises > 6 mm Hg was significant using the chi 2 test while the acetazolamide group showed a significant difference for rises > 6 and 10 mm Hg compared with the combined group. All acute pressure rises were recorded before or at 9 hours following operation except in the combined treatment patient where the rise occurred at 24 hours. To prevent the acute IOP rises seen following cataract surgery with lens implant in glaucoma patients we recommend combined ocular hypotensive therapy.


Tono-Pen tonometry in normal and in post-keratoplasty eyes. Oculab Tono-Pen tonometry was compared with Goldmann applanation tonometry in 82 eyes of 82 patients with normal corneas and in 54 eyes of 54 patients who had undergone penetrating keratoplasty and whose corneas did not preclude the use of Goldmann tonometer. We found that the intraocular pressure (IOP) in 48% of the eyes with normal corneas and in 57% after keratoplasty has different measurements with Goldmann and Tono-Pen pressures of 3 mm Hg or more. Despite the correlation between the Goldmann tonometer and the Tono-Pen in the group of eyes with normal corneas (r = 0.83) as well as in the group of eyes after keratoplasty (r = 0.79) the Tono-Pen tended to significantly overestimate the Goldmann tonometer reading (p < 0.0001). The mean difference between the two instruments was highest across the lower IOP range (< 9 mm Hg) in the group of eyes after keratoplasty. Because the mean absolute values of the paired differences between Goldmann and Tono-Pen measurements varied significantly across all IOP intervals it was not possible to establish a correction factor which could be used when comparing the two measurements. Based on this study the Tono-Pen consistently overestimated the actual IOP in an unpredictable manner. Where possible Goldmann measurements of the IOP are still to be preferred.


Visual evoked potentials after retrobulbar or periocular anaesthesia. The effect of the local anaesthetic agent, etidocaine, on the optic nerve function was examined at regional ophthalmic anaesthesia. Visual evoked potential (VEP) was recorded before and 15 minutes after injection of the anaesthetic agent in 19 patients scheduled for elective cataract surgery (seven retrobulbar and 12 periocular). Both the anaesthetised--that is, the eye to be operated on--and the fellow eye were examined. In the retrobulbar group, two patients displayed non-recordable VEPs while one had virtually non-detectable waves following the anaesthesia. In two retrobulbarly anaesthetised eyes, later peaks were unidentifiable while two other eyes had decreased amplitudes. In the periocular group, in nine patients, there was no clearcut effect on VEP resulting from the anaesthetic. In three patients of this group mild changes in the anaesthetised eyes were found. The differences in the effect of retrobulbarly or periocularly injected anaesthetics on VEP are probably due to the different concentration of the anaesthetic agent around the optic nerve.


Immunopathological findings in conjunctival cells using immunofluorescence staining of impression cytology specimens. The conventional technique of impression cytology provides a non-invasive method for the evaluation of conjunctival epithelium alterations. Using indirect immunofluorescence procedures two inflammatory markers, class II MHC antigens HLA DR and the receptor to IgE (CD23), were sought in impression cytology specimens obtained from 80 patients. In normal subjects conjunctival epithelial cells did not show any reactivity. Only scattered dendritic cells were found to express class II antigens but not the receptor to IgE. In contrast patients with chronic conjunctivitis of various aetiologies, mainly infectious or allergic, had 40-100% of brightly positive conjunctival cells for one or both antigens. In these cases epithelial cells and goblet cells reacted similarly. Twenty four eyes in 12 patients with idiopathic dry eye syndrome disclosed results similar to those from normal conjunctival specimens. However 18 other specimens from patients suffering from idiopathic tear deficiency but undergoing multiple substitutive treatments for dry eye had moderate to strong positivity for HLA DR and/or the receptor to IgE (20-100% of cells). As these results were independent of the degree of squamous metaplasia the expression of these membrane markers may reflect local inflammation in addition to tear deficiency, possibly due to sensitisation to the eye drops used. These immunocytological techniques thus provide useful methods of investigating conjunctival inflammation and allergy. They may constitute valuable aid in the diagnosis and appropriate treatment of ocular surface disorders.


Proliferative retinal diseases: myofibroblasts cause chronic vitreoretinal traction. Myofibroblasts are contractile cells having features intermediate between those of the fibroblast and smooth muscle. Myofibroblasts in preretinal membranes may cause retinal traction in proliferative retinal diseases. In this study, we demonstrate the presence of a chronically contractile subpopulation of myofibroblasts in pre-retinal membranes. Apha smooth muscle actin or desmin, both markers of the myogenically differentiated and chronically contractile subpopulation could be detected in a large majority of the examined membranes. Our findings may serve to explain the cellular basis of vitreoretinal traction in proliferative vitreoretinopathy and proliferative diabetic retinopathy.


Experimental autoimmune uveoretinitis in the RCS rat: the influence of photoreceptor degeneration on disease expression. S-antigen induced experimental autoimmune uveoretinitis (EAU) was produced in the Royal College of Surgeons (RCS) strain of rat which develops a photoreceptor dystrophy within 2 weeks of birth. Animals were sensitised at 60, 90, and 105 days of age: all animals developed disease, but onset was significantly delayed in older (105 day) animals compared with those aged 60 days at sensitisation (p 0.003). Disease was characterised by the early development of complete serous retinal detachment which resolved in a few days: the prevalence of retinal detachment was increased to 80% in dystrophic animals compared with 10% in the congenic, non-dystrophic controls (p < 0.001). Anterior uveitis was seen in 17/30 control strain eyes, but in none of 30 dystrophic eyes (p < 0.001). Genetically determined photoreceptor and retinal pigment epithelium dysfunction in the RCS rat, which may involve the local accumulation of altered S-antigen, predisposes the dystrophic strain to display an acute retinal detachment in the early stages of EAU. This phenomenon illustrates how biochemical dysfunction of a target organ may influence susceptibility, form, and severity of an experimental autoimmune disease.


Ciliary block (malignant) glaucoma after cataract extraction with lens implant treated with YAG laser capsulotomy and anterior hyaloidotomy. An 84-year-old woman developed high intraocular pressure with a shallow anterior chamber 2 months after an extracapsular cataract extraction with posterior chamber lens implant. The condition did not respond to peripheral iridectomy and removal of the implant but was treated successfully with YAG laser capsulotomy and anterior hyaloidotomy.


Congenital adenoma of the iris and ciliary body: light and electron microscopic observations. A 23-year-old man had a lesion in the right inferior iris which appeared to have enlarged since it was first seen when the patient was aged 5 years. The lesion was excised by a partial iridocyclectomy. Histopathologically the neoplasm was composed of both pigmented and non-pigmented cells. Pseudoacini, containing acid mucopolysaccharides, were present throughout the tumour matrix. Electron microscopically the non-pigmented cells were found to possess a convoluted plasmalemma, abundant rough endoplasmic reticulum, and numerous desmosomes and gap junctions. The pigmented cells contained large, round, mature melanosomes, occasional premelanosomes, and desmosomes, which resembled the posterior pigment epithelium of the iris. The intercellular matrix contained fine collagen fibrils resembling vitreous. We believe that this neoplasm represents a congenital adenoma of the ciliary body and iris.


Late onset Leber's optic neuropathy: a case confused with ischaemic optic neuropathy. A case is reported of a 63-year-old man with progressive central visual loss in one eye followed 11 months later by involvement of the fellow eye. A diagnosis of chronic ischaemic optic neuropathy was considered. However, despite a negative family history, the absence of electrocardiographic abnormalities, and minimal fundus changes a diagnosis of Leber's optic neuropathy was made on the basis of magnetic resonance imaging findings and the mitochondrial DNA mutation at base pair 11778.


Concurrent uveoretinitis and pineocytoma in a child suggests a causal relationship. Uveoretinitis was observed in a 9-year-old girl 6 months prior to the clinical appearance of a pineal tumour. Surgical removal was not successful but biopsy revealed a parenchymal neoplasm with differentiated pinealocytes and absent mitotic activity. Some of the tumour cells contained S-antigen, rhodopsin, and serotonin. Systemic glucocorticoid therapy followed by radiation therapy caused considerable reduction in size of the tumour and a complete normalisation of all eye symptoms. This report demonstrates for the first time that a pineocytoma can occur together with uveoretinitis in humans. The latter resembles the experimentally induced autoimmune uveoretinitis described in animals. It is speculated that the retinitis might reflect an autoimmune response to S-antigen present in some tumour cells of the pineocytoma.


Severe lower tracheal stenosis in infancy. 16 infants with severe distal tracheal stenosis (TS) seen from 1978-1988 are reviewed. Diagnosis was established by endoscopy and confirmed by radiographic methods, including magnetic resonance imaging (MRI) in three cases. Types of TS ranged from segmental stenosis to complex lesions involving the carina and bronchi. Associated vascular anomalies were found in nine patients (56%). All patients needed long-term intubation or a tracheostomy (seven cases) for stenting of the airway. Patients with congenital TS (CTS) were treated by tracheal resection (two survivors), dilatation (one survivor, two non-survivors), or observation (two non-survivors). Patients with pulmonary artery sling (PAS) were treated by division of the vascular sling (three survivors, one non-survivor), or combined vascular and tracheal surgery (one non-survivor). One patient with dilated pulmonary arteries compressing both main bronchi died in spite of oxygen therapy. Two cases of compression by an anomalous aorta were managed with dilatation (one survivor, one non-survivor). One patient with a severe stricture after a tracheal resection was successfully treated by dilatation. The overall mortality was 50%. We conclude that 1. endoscopic examination of the tracheobronchial tree and thorough evaluation of associated anomalies is mandatory in symptomatic patients; 2. if indicated surgical correction is the treatment of choice for tracheal stenosis; 3. dilatation and stenting of the stenosis are invaluable tools in the management of these infants.


Plasma concentrations of fat-soluble vitamins A and E in neonates with myelomeningocele. A group of newborn infants with myelomeningocele was compared with a group of neonates suffering from gastrointestinal malformations and a group of healthy children less than one year of age. The parameters studied were prealbumin, retinol-binding protein and the fat-soluble vitamins A and E. There were no significant differences between the groups in the plasma levels of the proteins or vitamin E. Vitamin A levels in newborn infants with myelomeningocele were found to be significantly lower compared to both the other groups.


Long-term results after partial and unilateral nephrectomy in childhood. In a clinical follow-up, 47 patients after unilateral or partial nephrectomy were examined. The average age at operation was 5.8 years (range 3 days-17.1 years), the average follow-up period after surgery was 11.9 years (range 8.2-18.3 years). Main indications for surgery were hydronephrosis (n = 27), other congenital malformations (n = 6) and neoplasia (n = 9). In one patient, we found hypertension, two patients presented with moderately diminished creatinine clearance, two further patients suffered from unilateral hydronephrosis after partial nephrectomy. We found by ultrasonography marked compensatory hypertrophy in cases of solitary kidneys (225.6 +/- 49.3%) as well as contralateral kidneys after partial nephrectomy (158.7 +/- 55.6%).


Predictability of duration and severity of congenital hydronephrosis as a cause of smooth muscle deterioration in pyelo-ureteral junction obstruction. Between 1989 and 1990, 31 new unilateral PUJO, prenatally diagnosed (16th-37th week of gestational age), underwent pyeloplasty between 19 and 105 days of age. The correlation between severity (V) (volume of hydronephrotic kidney) and duration (dT) of congenital hydronephrosis from its first detection in utero to surgery and histological findings were prospectively evaluated in this series. A dT longer than 70 days and a V larger than 50 ml., were considered as predictable for hypotrophy with fibrosis of the upper urinary tract smooth musculature. In conclusion, time seems to play an important role in the increase of the volume of the obstructed urinary tract to the extent of a large-scale hydronephrosis.


20 years of functional reconstructive surgery in bladder exstrophy--balance 10 years after a preliminary report. This is an update of a consecutive series of 19 patients with bladder exstrophy presented as a preliminary report 10 years ago with a follow-up of 6 years. The average time of follow-up now is 14 years. In difference to the former report several additional procedures have been performed in most patients. 3 early "good" patients stay "good and continent". Only 3 out of 5 patients categorized as "fair" are now "good and continent" and only one without further intervention. 3 out of 6 patients who were classified as "poor" are now diverted. The overall rate of secondary diversions is 40% including 2 patients with ureterosigmoidostomies. All patients have normal renal function and normal blood pressures. These results show that an early "good" result generally remains "good" but additional surgery may be warranted. The initially "fair" results usually necessitate a secondary procedure, e.g. augmentation cystoplasty or/and bladder neck tightening to show an improvement. Patients with early "poor" results unfortunately remain "poor" and have a great chance to get diverted.


Hyperplastic callus formation in osteogenesis imperfecta. Osteogenesis imperfecta, an inherited disorder of connective tissues, affects roughly (OI) 4000 people in Germany (11). The main clinical symptoms are fragile bones, progressing skeletal deformities, generalized osteoporosis and short stature. Incidentally, the clinical manifestations can range from perinatal lethal forms to phenotypical normal adults. In many instances the underlying causes of the disease are mutations in gene coding for collagen I, the predominant protein in most connective tissues. Fracture healing is usually not impaired, although in a unique group of OI-patients, a tumor-like hyperplastic callus occurs with excessive deposition of extracellular matrix constituents. Biochemical analysis of the callus is reminiscent of bone from early stages of human development and normal fracture healing (e.g. collagen type composition, degree of posttranslational modification). This underlines that, besides collagen mutations, the regulation of collagen synthesis and their posttranslational processing might be disturbed in patients with hyperplastic callus formation.


Does fibrin glue cause foreign body reactions? The study was performed in 20 Wistar rats. Approximately 4 mm perforations were created in the left colon of ten rats to which fibrin glue was applied (Group D). The control group had the same injury without any therapeutic or surgical measures (Group K). Of the 20 rats in both groups none died during the postoperative period. All animals sacrificed on the 11th postoperative day. All ten rats of Group D displayed inflammatory foreign body reactions. The control group showed no such reaction.


Manometric evaluation of children with chronic constipation using a suction-stimulating electrode. Thirteen children with chronic constipation (8 males and 5 females, aged 1 month to 9 years) were evaluated manometrically. A suction-stimulating electrode was used to induce the rectoanal reflex electrically so as to examine the threshold voltage and latency of this reflex. The rectoanal reflex was induced in all patients by balloon dilatation of the rectum. However, electrical stimulation of the rectal mucosa triggered a reflex in only nine of them and the threshold voltage and latency was 22.2 +/- 3.6 volts (mean +/- SD, n = 9) and 2.04 +/- 0.38 seconds (n = 9), respectively. Hence, it is suggested that a suction-stimulating electrode is a useful supportive tool in manometric anorectal studies, and that disturbances in the rectal nervous system might be one of the causative factors in chronic constipation in childhood.


Rupture of the duodenum with avulsion of the papilla of Vater due to blunt trauma in a child, and review of the literature. Duodenal rupture with avulsion of the papilla of Vater is an uncommon injury due to blunt abdominal trauma in children. This report describes a 9-year-old boy who sustained this injury and was treated successfully two days delayed by a Roux-en-Y choledocho-pancreaticojejunostomy and temporary implantation of a zipper for abdominal wall closure. The rarity of this complex injury and its initial presentation as a posttraumatic pancreatitis warrant its description. A review of the literature, modes of diagnosis and surgical technique for repair are described.


Acute acalculous cholecystitis caused by Salmonella typhi in a 6-year-old child. A rare case of acute acalculous cholecystitis caused by Salmonella typhi in a 6-year-old child is presented. The clinical signs were fulminant, with diffuse peritonitis being suspected. Cholecystostomy and i.v. ceftriaxone proved efficacious and the girl was discharged in less than two weeks. The appropriate literature is reviewed.


Penile dorsal angulation in a child without epispadias. This is a report on a 5-year-old boy with dorsal penile angulation without epispadias. The surgical correction is described.


Polyorchidism: a case report and review of the literature. We report on a case of polyorchidism associated with left-sided inguinal hernia and hydrocele. Biopsies of both testes revealed normal histological pattern. After the hernia repair and hydrocelectomy both testes were left at their original places in the scrotum and fixed. Polyorchidism and its management are discussed briefly.


Thermography in a child with varicocele. A 12-year-old boy with a left-sided varicocele (Grade 3) was assessed preoperatively and followed postoperatively by computer-assisted infrared thermography. The temperature of the left scrotum was 35.4 degrees C before surgery which was 4 degrees C higher than that of the right scrotum. After ligation of the left internal spermatic vein and removal of the varicocele, scrotal thermograms 39 days and 12 months after operation were normal. Therefore, it seems that thermography is very valuable in the evaluation of testicular function in children with varicocele, in whom semen analysis is impossible.


Reconstruction of the perineal body after recurrence of a recto-perineal fistula in an intermediate type of anorectal malformation in a young woman. The appearance of a recto-perineal fistula post-operatively in an intermediate type of anorectal malformation is rare. It always gives rise to incontinence. However, the closure of this fistula is particularly difficult since recurrence is frequent. The following case report concerns a patient who after several unsuccessful attempts to close the fistula, had a local rotation flap which permitted good healing and a beneficial effect on continence thanks to a good reconstruction of the perineal body.


Further characterization of the acetyl LDL (scavenger) receptor expressed by rabbit smooth muscle cells and fibroblasts. We have previously demonstrated that the acetyl low density lipoprotein (LDL), or scavenger, receptor is expressed by rabbit smooth muscle cells (SMCs) and fibroblasts. Moreover, receptor activity in rabbit fibroblasts was regulated over a wide range by preincubating the cells with secretion products from human platelets or with phorbol esters. The current studies were undertaken to examine the regulation of receptor expression in rabbit SMCs, to characterize further the receptor expressed by rabbit SMCs and fibroblasts, and to determine whether incubating these cells with chemically modified lipoproteins would lead to foam cell formation. Receptor activity was increased fivefold in rabbit SMCs by preincubation with platelet secretion products and nine- to 20-fold by preincubation with phorbol esters or mezerein, a non-phorbol activator of protein kinase C. The phorbol ester-induced increase in receptor activity was due to an increased mass of scavenger receptor as determined by both ligand blots and immunoblots of membrane proteins from these cells. The immunoblotting studies suggest that the SMC scavenger receptor activity is due to type I or type II scavenger receptors. The scavenger receptor expressed by phorbol ester-treated rabbit SMCs and fibroblasts bound chemically modified LDL with an order-of-magnitude higher affinity (Kd 5.1 x 10(-10) M) than did the scavenger receptor expressed by mouse peritoneal macrophages (Kd 4.1 x 10(-9) M), whereas rabbit macrophages bound chemically modified LDL with intermediate affinity (Kd 1.0 x 10(-9) M). Both the phorbol ester-stimulated rabbit fibroblasts and SMCs expressed 10-30% as many receptors per cell as did mouse peritoneal macrophages. Consistent with the difference in the number of receptors, uptake of the chemically modified LDL at 37 degrees C by SMCs led to approximately 25% as much lysosomal degradation and stimulation of cholesterol esterification as in mouse peritoneal macrophages. Incubation of phorbol ester-treated rabbit fibroblasts or SMCs with chemically modified LDL (50 micrograms/ml) for 24 or 48 hours, respectively, resulted in a threefold increase in total cholesterol and a 15-fold increase in cholesteryl ester within the cells. These studies suggest that the scavenger receptor-mediated uptake of modified lipoproteins may contribute to the formation of SMC foam cells in atherosclerotic lesions.


Experimental atherosclerosis at the carotid bifurcation of the cynomolgus monkey. Localization, compensatory enlargement, and the sparing effect of lowered heart rate. We have characterized plaque localization, the extent of compensatory artery enlargement, and the effect of heart rate in experimental atherosclerosis at the carotid bifurcation of the cynomolgus monkey. We altered heart rate by sino-atrial node ablation (SNA) and then fed the animals an atherogenic diet for 6 months. Heart rate was measured at four time points by 24-hour telemetry. Of nine animals with SNA, heart rate was reduced significantly in six (from 148 +/- 11 to 103 +/- 20 beats/min, p < 0.001) and was unchanged in three. Sham-operated monkeys had no significant change in heart rate. On the basis of comparison with the preoperative mean for all 17 animals (136 +/- 22 beats/min), animals were separated into a low-heart-rate (LHR) group (111 +/- 16 beats/min, n = 12) and a high-heart-rate (HHR) group (150 +/- 16 beats/min, n = 5). Blood pressure, serum cholesterol level, and body weight did not differ for the two groups. As in the human, plaques formed predominantly in the proximal portion of the internal carotid artery at the lateral wall opposite the flow divider. Plaque cross-sectional area increased progressively from the relatively uninvolved, adjacent common carotid artery to the mid-sinus region of the internal carotid artery and decreased from the mid-sinus region to the internal carotid artery beyond the sinus. Plaque distribution was the same for the LHR and HHR groups, but lesion area and percent stenosis were greater for the HHR group than for the LHR animals (2.01 +/- 1.19 compared with 0.76 +/- 0.42 mm2 for lesion area [p < 0.02] and 30.7 +/- 4.4% compared with 15.2 +/- 7.3% for stenosis [p < 0.002]).(ABSTRACT TRUNCATED AT 250 WORDS)


Vascular endothelium responds to fluid shear stress gradients. In vitro investigations of the responses of vascular endothelium to fluid shear stress have typically been conducted under conditions where the time-mean shear stress is uniform. In contrast, the in vitro experiments reported here have re-created the large gradients in surface fluid shear stress found near arterial branches in vivo; specifically, we have produced a disturbed-flow region that includes both flow separation and reattachment. Near reattachment regions, shear stress is small but its gradient is large. Cells migrate away from this region, predominantly in the downstream direction. Those that remain divide at a rate that is high compared with that of cells subjected to uniform shear. We speculate that large shear stress gradients can induce morphological and functional changes in the endothelium in regions of disturbed flow in vivo and thus may contribute to the formation of atherosclerotic lesions.


Monoclonal antibodies against LDL further enhance macrophage uptake of LDL aggregates. Self-aggregates of low density lipoprotein (LDL) are taken up and degraded more rapidly by macrophages than is native LDL. That enhanced uptake is attributable in part to phagocytosis via the LDL receptor pathway. However, arterial macrophages appear to express little LDL receptor activity. The present studies demonstrate an alternative mechanism by which LDL aggregates could contribute to foam cell formation. This could occur by the formation of large immune complexes that are taken up by macrophages via the Fc receptor. When immune complexes were formed with native, soluble LDL and MB47, a monoclonal antibody specific to the apoprotein B domain recognized by the LDL receptor, the subsequent uptake and degradation of the LDL by macrophages were inhibited 50-80% compared with native LDL alone. In contrast, when aggregated LDL was bound to MB47 at a similar molar ratio, the subsequent degradation of the insoluble immune complexes was two- to fivefold greater than that of aggregated LDL alone. The enhanced uptake was abolished when Fab or F(ab')2 fragments of MB47 were substituted for the intact antibody, indicating that the increased uptake was via the Fc receptor pathway. Furthermore, the uptake of the immune complexes of aggregated LDL was reduced by competition for the Fc receptor with heat-aggregated immunoglobulin. There was also an increase in the rate of cellular cholesterol esterification and an increase in macrophage cholesteryl ester mass. Since aggregates of LDL as well as autoantibodies against modified LDL have been demonstrated in atherosclerotic lesions, it is possible that immune complexes of aggregates of modified LDL may be generated in the intima.(ABSTRACT TRUNCATED AT 250 WORDS)


Time course and cellular characteristics of the iliac artery response to acute balloon injury. An angiographic, morphometric, and immunocytochemical analysis in the cholesterol-fed New Zealand white rabbit. Evaluation of the response of the arterial vessel wall to acute arterial injury in experimental models has taken on substantial importance because of an increasing interest in angioplasty treatment of human atherosclerotic lesions. In this study, the response of normal arterial vessels to acute balloon injury was studied in 45 iliac artery segments from 24 New Zealand White rabbits fed a 2% cholesterol diet. At specified time points between 1 and 41 days after the initial balloon pullback injury, the iliac arteries were analyzed by angiographic, morphometric, and immunocytochemical techniques. Angiographic measurements indicated progressive compromise of the iliac artery lumen with increasing duration of time from injury. Morphometric measurements showed that intimal area increased from 0.004 +/- 0.01 mm2 3 days after injury to 1.15 +/- 0.30 mm2 34-41 days after injury. Cell line-specific immunocytochemical analysis identified the macrophage as a prominent component of the earliest intimal cellular infiltrate. Smooth muscle cells appeared within the intima 7-9 days after injury. As the intima increased in area, macrophages predominated along the internal elastic lamina aspect of the intimal lesion while smooth muscle cells occupied the portion of the intima adjacent to the lumen. In summary, retrograde balloon pullback injury followed by cholesterol feeding results in progressive arterial luminal narrowing due to a progressively enlarging intimal cellular infiltrate. The temporal and spatial contributions of smooth muscle cell and macrophage components of the developing intimal cellular infiltrate have been characterized.


Hepatic ACAT activity in African green monkeys is highly correlated to plasma LDL cholesteryl ester enrichment and coronary artery atherosclerosis. Previous studies and this study of African green monkeys show a strong positive correlation between plasma low density lipoprotein (LDL) size and the extent of coronary artery atherosclerosis (CAA). Increased LDL size was principally due to the accumulation of cholesteryl oleate molecules within the particle core, suggesting that many of these cholesteryl esters were of tissue origin, i.e., from the acyl-coenzyme A:cholesterol acyltransferase (ACAT) reaction instead of the lecithin:cholesterol acyl-transferase (LCAT) reaction. The current study was conducted to test the hypothesis that ACAT in the liver is the source of the increased numbers of cholesteryl oleate molecules in plasma LDL particles that appear to increase the atherogenic potential of LDL. Monkeys were fed diets rich in fat (lard, safflower oil, or fish oil) and cholesterol for 3-6 years before liver perfusion, ACAT assay, and evaluation of CAA. Hepatic ACAT activity was positively correlated with hepatic cholesteryl ester secretion (r = 0.61, p < 0.001), plasma LDL cholesteryl ester content (r = 0.60, p < 0.0001), and the extent of CAA (r = 0.62, p < 0.0001). The number of cholesteryl oleate molecules within LDL increased proportionally with LDL size in each of the diet groups. Hepatic cholesteryl oleate concentration was correlated with the accumulation of cholesteryl oleate in liver perfusate (r = 0.72, p < 0.01) and with plasma LDL cholesterol oleate content (r = 0.73, p < 0.0001). Our interpretation is that these data, obtained in a relevant primate model of CAA, suggest that hepatic ACAT increases the atherogenicity of LDL by augmenting both the secretion by the liver and accumulation in plasma LDL of cholesteryl oleate.


Change in LDL particle size is associated with change in plasma triglyceride concentration. Low density lipoprotein (LDL) particle size is inversely associated with plasma triglyceride concentration in cross-sectional analyses. In the present study, changes in the LDL particle size of 227 participants of the Framingham Offspring Study were analyzed longitudinally by nondenaturing gradient gel electrophoresis at two examinations that were separated by 3-4 years. All subjects had triglyceride concentrations < 400 mg/dl at both exams. Using laser scanning densitometry to assess mean LDL particle size, 56% of samples displayed a change in size: 41% had a one-band size change, 13% had a two-band change, and 2% had a three-band change. These changes in size corresponded to a 15% change in pattern type, based on pattern A and B terminology. There was a significant inverse association between change in LDL size and change in triglyceride (p < 0.0001) and glucose (p < 0.004) concentrations, body weight (p < 0.02), and age (p < 0.03). There was also a significant positive association with change in high density lipoprotein (HDL) cholesterol concentration (p < 0.0001). Change in LDL cholesterol concentration, as calculated by use of the Friedewald formula, however, showed no significant association with change in LDL size (p < 0.9). There was also no significant association with change in smoking or blood pressure, but there was a nonsignificant inverse trend associated with alcohol intake (p < 0.08).(ABSTRACT TRUNCATED AT 250 WORDS)


Cholesterol efflux is defective in macrophages from atherosclerosis-susceptible White Carneau pigeons relative to resistant show racer pigeons. White Carneau (WC) pigeons are susceptible to the development of aortic atherosclerosis, whereas Show Racer (SR) pigeons are resistant, even though there are no differences in the known risk factors, including plasma cholesterol levels, lipoproteins, blood pressure, etc. Although this suggests that the difference in atherosclerosis susceptibility between WC and SR pigeons may be mediated at the level of the arterial wall, we have yet to identify a mechanism that can account for this difference. In pigeons as in other species (including humans), macrophages play a major role in the pathogenesis of atherosclerosis. Pigeon macrophages have multiple mechanisms for the uptake of lipoproteins and the accumulation of cholesteryl esters. To date, however, no differences in lipoprotein uptake between macrophages of WC and SR pigeons have been identified that could explain the difference in atherosclerosis susceptibility. In the present study we explored the alternative hypothesis that there are differences in the rate of cholesteryl ester clearance from peritoneal macrophages isolated from the two breeds of pigeons. Cholesterol efflux studies were conducted with elicited pigeon peritoneal macrophages that were loaded with cholesteryl ester either in vitro by incubation with rabbit beta-very low density lipoprotein or in vivo by isolation of macrophages from birds fed a cholesterol-containing diet. Using these techniques we were able to load WC and SR macrophages consistently with cholesteryl esters to levels typical of arterial foam cells (150-1,150 micrograms/mg cell protein). Under these cholesterol loading conditions there was no net efflux of cholesterol from either WC or SR macrophages when incubated for up to 24 hours in the presence of pigeon or human high density lipoprotein (HDL), fetal bovine serum, or lipoprotein-deficient serum. Under the same conditions, efflux of cholesterol from mouse peritoneal macrophages was stimulated by human and pigeon HDL. Despite the inability of HDL, lipoprotein-deficient serum, or fetal bovine serum to promote net cholesterol efflux, apoprotein (apo) HDL/phosphatidylcholine (PC) vesicles stimulated cholesteryl ester clearance from both WC and SR pigeon macrophages but at a significantly slower rate from WC pigeon macrophages. When incubated in the presence of excess apoHDL/PC (400 micrograms/ml), the rate of depletion of cellular cholesteryl esters was log-linear for at least 48 hours. WC macrophages cleared an average of only 9% of their cholesteryl esters in 24 hours when incubated with excess apoHDL/PC, whereas SR macrophages reduced their cholesteryl ester content by an average of 42%.(ABSTRACT TRUNCATED AT 400 WORDS)


Long-term nicotine exposure increases aortic endothelial cell death and enhances transendothelial macromolecular transport in rats. Repeated endothelial cell injury has been suggested as an initiating factor in atherogenesis. Dying or dead endothelial cells have been shown to make significant contributions to the local enhancement of transendothelial macromolecular transport. Since cigarette smoking is one of the major risk factors for atherosclerosis, we examined the hypothesis that smoking accelerates atherogenesis by increasing the frequency of endothelial cell death and hence transendothelial macromolecular transport. Sixteen male Sprague-Dawley rats were given nicotine at a weight-adjusted dose of 5 mg/kg body wt per day in their drinking water over a period of 6 weeks. A group of 16 age-matched male Sprague-Dawley rats not exposed to nicotine and maintained over the same time period served as the control group. In en face preparations of thoracic aorta, immunoglobulin G-containing dying or dead endothelial cells were identified by the indirect immunoperoxidase method, and endothelial leakage to Evans blue-albumin (EBA) complexes (5 minutes after intravenous injection) was visualized by fluorescence microscopy. The results showed that in nicotine-treated rats, 51% of dead endothelial cells were associated with EBA leakage, which was responsible for 57% of total EBA leaky foci. Both the frequency of endothelial cell death (0.94 +/- 0.11% versus 0.40 +/- 0.04%, p < 0.0001 by two-tailed, unpaired Student's t test) and the number density of EBA leaky foci (6.45 +/- 1.23/mm2 versus 3.30 +/- 0.49/mm2, p < 0.05 by two-tailed, unpaired t test) were significantly greater in nicotine-treated rats than in control rats.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of endothelial integrity, transmural pressure, and time on the intimal-medial uptake of serum 125I-albumin and 125I-LDL in an in vitro porcine arterial organ-support system. This report describes a new in vitro, metabolically supported, Sinclair Research Farm minipig aortic preparation in which the intimal-medial uptakes (M, mg.cm-2 of intimal surface) of porcine 125I-albumin and normocholesterolemic (nonoxidized) porcine 125I-low density lipoprotein (LDL) from a stirred, autogenous serum (containing a 125I-protein concentration of c0, mg.cm-3 at 37 degrees C and pH 7.4) were studied as functions of transmural pressure (0 < or = P < or = 150 mm Hg), time (30 < or = t < or = 120 minutes), and endothelial integrity. The following new observations were made: 1) The normalized transendothelial uptakes (M/c0, cm) of both albumin and LDL across normal intact aortic endothelial surfaces were insensitive to P. This indicated that these macromolecular solutes were not readily convected across the normal aortic endothelial surface despite increasing P. 2) However, the associated transendothelial M/c0 versus t relations for the normal intact surfaces were shown to increase monotonically with t in a manner consistent with a simple diffusive transport across a large surface barrier into the subjacent media, either with (Cases 2A and 2B) or without (Case 1) an associated transmural water convection. 3) The shapes of these temporal M/c0 curves of albumin and LDL were virtually the same; however, the magnitude of the albumin M/c0 curve was about sevenfold greater than that of LDL. 4) The M/c0 across the injured endothelial surface (Case 2C) not only increased monotonically with t but also increased significantly with P, indicating that in the absence of a normal endothelial surface, a very large convective component was added to the transport processes across the exposed aortic endothelial basement membrane and internal elastica. We conclude that: 1) the normal porcine aortic endothelial surface can provide a virtually complete barrier to the transendothelial convective transport of both albumin and LDL, 2) the diffusive barrier of the normal endothelial surface to LDL was sevenfold greater than that to albumin, 3) loss of the endothelial cell layer was associated with a threefold increase in the (P = 0) diffusive intimal-medial uptake of serum albumin in contrast to an eightfold increase in the pressurized (P = 150 mm Hg) combined diffusive-convective intimal-medial albumin uptake in the same vessel.


Mast cell-mediated inhibition of reverse cholesterol transport. Net cholesterol efflux from cholesterol-loaded macrophages, i.e., foam cells, was induced by incubating the foam cells with high density lipoprotein3 (HDL3). However, when the incubation system included rat serosal mast cells stimulated to trigger exocytosis of their cytoplasmic secretory granules, the ability of HDL3 to induce cholesterol efflux was largely lost. This loss was found to be due to the proteolytic action of chymase, the neutral protease of the granules, which degraded the apolipoproteins of HDL3, so rendering them unable to mediate cholesterol efflux from the foam cells. The observation defines a novel cell-dependent mechanism that blocks the initial steps of reverse cholesterol transport and suggests a role for mast cell chymase in cellular accumulation of cholesterol, an early stage in atherogenesis.


Relation of triglyceride metabolism and coronary artery disease. Studies in the postprandial state. The status of fasting triglycerides as a risk factor for coronary artery disease (CAD) has been considered weak because in multivariate analyses, triglycerides tend to be eliminated by high density lipoprotein (HDL) cholesterol. To further evaluate the role of triglycerides in CAD, we employed postprandial lipemia as a more informative means of characterizing triglyceride metabolism. In 61 male subjects with severe CAD and 40 control subjects without CAD as verified by angiography, we measured cholesterol; triglycerides; HDL cholesterol; HDL2 cholesterol; and apolipoproteins A-I, A-II, and B in fasting plasma and triglycerides before and 2, 4, 6, and 8 hours after a standardized test meal. Both the maximal triglyceride increase and the magnitude of postprandial lipemia (area under the triglyceride curve over 8 hours after the meal) were higher in cases than in control subjects. Single postprandial triglyceride levels 6 and 8 hours after the meal were highly discriminatory (p < 0.001), and by logistic-regression analysis displayed an accuracy of 68% in predicting the presence or absence of CAD. In this respect, accuracy was higher than that of HDL2 cholesterol (64%) and equal to that of apolipoprotein B (68%), the most discriminatory fasting parameter. Multivariate logistic-regression analysis was performed to reduce the number of risk factors to those that were statistically independent. This statistical procedure selected postprandial but not fasting triglycerides into the most accurate multivariate model, which also contained the accepted risk factors HDL2 cholesterol, apolipoprotein B, and age. This model classified 82% of subjects correctly. We conclude that triglycerides are independent predictors of CAD in multivariate analyses including HDL cholesterol, provided that a challenge test of triglyceride metabolism such as postprandial lipemia is used. The study suggests that the metabolism of triglycerides is a critical determinant of cholesterol metabolic routing. The findings support the concept that the negative association between HDL cholesterol levels and CAD actually originates in part from a positive relation between CAD and plasma triglycerides, as ascertained in the postprandial state.


Serum Lp(a) as a discriminant marker of early atherosclerotic plaque at three extracoronary sites in hypercholesterolemic men. The PCVMETRA Group. To investigate the role of lipoprotein (a) (Lp[a]) as an atherogenic condition related to hypercholesterolemia, we studied the serum concentration of Lp(a) as measured by immunonephelometry in relation to the presence of asymptomatic echographic plaques in the peripheral arteries of 103 untreated hypercholesterolemic, normotensive, middle-aged men. Plaque was found at carotid, aortic, and femoral sites in 36%, 51%, and 53% of subjects, respectively. The Lp(a) level was higher in the group with carotid plaques than in the group without (0.29 +/- 0.20 versus 0.17 +/- 0.14 g/l, p < 0.01), not significantly higher in the group with aortics plaque than in the group without (0.24 +/- 0.19 versus 0.19 +/- 0.16 g/l), and not different between groups with and without femoral plaques (0.21 +/- 0.18 versus 0.22 +/- 0.17 g/l). A logistic regression analysis confirmed that Lp(a) was associated with carotid plaques (p = 0.004), independent of other risk factors. However, in patients with low density lipoprotein cholesterol values above the group median value (4.7 mmol/l), Lp(a) was associated not only with carotid plaques (p < 0.01) but also with aortic plaques (p < 0.05), as well as with the number of diseased sites (p = 0.02). In contrast, in patients with low density lipoprotein cholesterol levels below or equal to 4.7 mmol/l, Lp(a) only remained associated with carotid plaques (p < 0.05). Thus, in symptom-free, hypercholesterolemic men, early atherosclerosis was influenced by serum Lp(a), particularly in the carotid arteries, as well as by the presence of a higher level of low density lipoprotein cholesterol.


LDL binding sites on platelets differ from the "classical" receptor of nucleated cells. Washed human platelets bound radioiodinated low density lipoprotein (125I-LDL) to a class of saturable binding sites; they numbered 1,348 +/- 126 per platelet, and the dissociation constant (KD) was 50.7 +/- 9 nM. 125I-LDL binding to platelets was reversible, and apparent equilibrium was attained within 25 minutes at 22 degrees C and was characterized by forward and reverse rate constants of 1.47 x 10(4) x sec-1 x M-1 and 8 x 10(-4) x sec-1 x M-1, respectively. Such binding was largely unaltered by temperature, divalent ions, and chelating agents. In addition, neither did receptor regulation (up or down) occur when platelets were loaded with cholesterol, nor did prostaglandin E1 (PGE1) increase the binding of 125I-LDL to platelets. On the other hand, the specificity of LDL binding was not typical of the LDL receptor of nucleated cells. Lipoproteins competed for the occupancy of LDL binding sites in platelets with the following order of potency: very low density >> intermediate density > high density subfraction 2. High density lipoprotein subfraction 3, heparin, and PGE1 had no effect on this binding. 125I-LDL binding to lymphocytes and fibroblasts and proteolytic degradation of 125I-LDL by lymphocytes was inhibited by the monoclonal antibody IgG-C7 directed against the LDL receptor to 88%, 85%, and 85% (p < 0.001), respectively. However, with this monoclonal antibody, a blocking effect on neither 125I-LDL binding to platelets nor on LDL-enhanced platelet aggregation induced by ADP and collagen was found. Moreover, we confirmed the existence of LDL binding in platelets from patients with familial hypercholesterolemia. Our results indicate that human platelets bind LDL by saturable sites, which clearly differ from the "classical" LDL receptor in their binding properties, absence of receptor regulation, presence in platelets of familial hypercholesterolemia patients, and the lack of a blocking effect of IgG-C7 on LDL binding and LDL biological activity.


Etoposide treatment suppresses atherosclerotic plaque development in cholesterol-fed rabbits. To study the mechanisms by which monocytes/macrophages and smooth muscle cells contribute to atherosclerotic lesions, we studied atherosclerotic plaque formation in cholesterol-fed rabbits treated with etoposide, a drug that has been shown to have several effects that could interfere with the proposed interactions between these two cell types (M.W. Aarnoudes et al, Virchows Arch B 1984;47:211-216 and M. Rozencweig et al, Cancer 1977;40:334-342). Our results show that long-term etoposide treatment of New Zealand White rabbits maintained on a high-cholesterol diet decreases the extent of fatty streak formation in the aortic intima. Moreover, the plaques formed in the presence of etoposide are thinner and at least focally have less fibrous tissue and fewer smooth muscle cell-derived foam cells than do plaques in control rabbits. These effects are independent of the extent of the diet-induced hyperlipemia or an effect of etoposide on blood cell count and may be related to the inhibition of intimal cell proliferation by etoposide.


Dentigerous cysts involving permanent incisors: four case reports. Four cases of dentigerous cysts involving permanent incisors are reported. In each case the primary predecessor had been traumatized, was discoloured and suspected of being non-vital. Epithelial proliferation of the follicular tissues induced by chronic periapical inflammation of the predecessors is suggested as the most likely pathogenesis of cystic transformation in these cases. Regular radiographic review of discoloured primary incisors is recommended.


A longitudinal study of the effects on the oral mucosa of treatment for acute childhood leukaemia. A longitudinal study was undertaken of the effect of therapy on the oral mucosa of 12 children with acute leukaemia. The major oral problem was ulceration, which was associated with neutrophil counts of less than 1.0 x 10(9)/l. Candida carriage rates were consistently high in patients with or without oral candidosis. Salivary flow was not significantly different between leukaemic children and the control group.


Dental health and salivary Streptococcus mutans levels in a group of children with heart defects. One-hundred children aged 2-16 years who were attending the Outpatients Department of the Yorkshire Regional Cardiac Centre were examined for dental caries, gingivitis, plaque and calculus. A control group of 100 children was also examined. A saliva sample was taken from each child in the study group to assess the level of Streptococcus mutans colonization. Comparing study and control groups, dental caries experience (dmft) was significantly higher only in the primary teeth of 5-9-year-old children in the study group, and there were no significant differences in gingivitis, plaque or calculus. The S. mutans count was found to be positively correlated to the number of decayed teeth in the study group.


Dental caries in 12-year-old children in Salford, UK: changes over 28 years. A sample of 111 12-year-old children in Salford, UK was examined in 1988, and the findings compared with those of a survey carried out in 1960 of 12-year-old children in the same city. Between 1960 and 1988, the mean DMFT of the boys fell from 6.04 to 2.34 and that of the girls from 6.54 to 3.40. All types of teeth were affected by caries in 1960 but in 1988 none of the canines and mandibular lateral incisors were affected. In 1960 the permanent first molars contributed 57% of the boys' and 54% of the girls' total DMFT while in 1988 they contributed 87% of the boys' and 73% of the girls' DMFT. Jackson's Treatment Index showed that the dental services were making a greater impact in 1988; for the boys it rose from 29.5 to 63.2 and for the girls from 27.8 to 62.3.


Flow rate and chemistry of parotid saliva related to dental caries and gingivitis in patients with thalassaemia major. Twenty-one child patients with thalassaemic major (TM) and 83 healthy control children were examined for dental caries and gingivitis. Stimulated parotid gland secretions were collected from each child. Parotid saliva flow rate was measured and the saliva samples were tested for calcium, phosphorus, potassium, sodium, urea, lysozyme and immunoglobulin levels (IgA, IgG, IgM). The results showed that dental caries experience was significantly higher in the TM group. Parotid saliva flow rates in TM patients were not significantly different from those in the healthy controls. However, the median saliva concentrations of phosphorus and IgA were significantly lower in the patients than in the controls. The concentration of lysozyme was also lower in the TM group, but the difference was not statistically significant. The findings could provide an explanation for the higher dental caries experience and gingivitis observed in the TM group.


Dental management of a child with EEC syndrome. The EEC syndrome is a condition characterized by ectrodactyly, ectodermal dysplasia and clefting of the lip and/or palate. Here we report a case of a boy with the EEC syndrome. The clinical and radiologic aspects of this condition, as well as the dental treatment, are reviewed.


[Significance of the so-called symptomless periods of disease]. Considerable change has occurred to nosology due to the advent of electron microscopy and its use in investigations of pathological processes. These investigations have shown that morphological alterations under the impact of pathogenic factors may persist for extended periods of time without any clinical manifestation due to soon effectiveness of mechanisms of compensation in the form of regeneration and hyperplasia of cells and intracellular ultrastructures (intracellular regeneration). Clinical symptoms will not grow manifest until regeneration and hyperplasia are not capable any more of compensation for tissue lesions that have resulted from dystrophy and necrosis. This is likely to offer an explanation for delay of treatment, when a patient will not see a physician until the disease has become incurable. It is the established purpose of early diagnosis to detect structural alterations in organs during the asymptomatic preclinical period, that is before the disease has reached an advanced stage. The idea of "primary existent functional and secondarily added morphological alterations" is just as erroneous as that of the existence of "purely functional diseases". Such outdated concepts used to be based on underestimation of compensatory reactions of the organism and the enormous role played by them in the development and outcome of pathological processes.


[Pathology of the placenta. XI. Feto-fetal transfusion syndrome]. An account is given in this part of the series of placental alterations in the foeto-foetal transfusion syndrome of monochorial twin pregnancies. Arteriovenous or less often arterio-arterial anastomoses between the two parts of the placenta were found to be conditions for such foeto-foetal transfusion. Blood is discharged on the foetal side from one twin to the other, with one of the twins being adversely affected in growth and general development (donor), which may be as bad as acardia in case of very early onset of transfusion. The other twin (recipient) will undergo better development but will as well be impaired by cardiac hypertrophy and other organic lesions due to excessive inflow of blood. The placenta of the donor--quite often meeting intra-uterine death--usually is smaller than that of the recipient. It is severely anaemic and, additionally, may be of inadequate maturity. The placenta of the recipient may exhibit strongly pronounced hyperaemia, up to A and B angiomatosis. The author describes two of his own cases, with reference being made to morphological findings.


[Pathomorphology of experimental Candida mycosis]. An account is given in this paper of evaluated results obtained from over 30 years of research on the pathogenesis and morphology of experimental candidiasis. No success has so far been recorded from efforts to set up a fully valid animal experimental model of all forms of candidiasis in man. Models so far can be successfully constructed only for certain links in the process of infection. Particular importance, in this context, should be attributed to effects on the organism which would cause improvement or reduction of specific and unspecific resistance. Critical analysis of results obtained from differently conceptualized experiments has contributed to comprehensive understanding of the morphogenesis of opportunistic candida infections in man. Models established in the wake of animal experiments have provided a basis for formulation of principles of a combined candidiasis therapy, using both antibiotics and immunomodulators.


Analysis of expression of erythropoietin-binding sites in human lung carcinoma by the biotinylated ligand. Recombinant glycosylated erythropoietin (EPO) was biotinylated with biotin-aminocaproyl hydrazide via periodate-treated sialic acid moieties and applied to sections of 64 tumors of the lower respiratory tract, comprising 19 primary adenocarcinomas, 19 epidermoid carcinomas, 13 large cell anaplastic carcinomas, 11 small cell lung carcinomas, 11 intrapulmonary metastases, 1 mesothelioma and 1 lymphocytic interstitial pneumonia. The formalin-fixed, paraffin-embedded specimens were incubated with labelled EPO at room temperature and a concentration of 10 micrograms/ml for 60 min. The expression of the EPO-binding sites was visualized by the ABC technique. All of the analyzed large cell anaplastic carcinomas and the majority of the epidermoid carcinoma (89%), adenocarcinoma (79%), and metastases (82%) displayed binding capacities for EPO. Five out of the eleven small cell lung carcinomas, the analyzed mesothelioma and lymphocytic interstitial pneumonia revealed definite staining, too. Binding sites could, in addition, be seen in air dried, non-fixed, acetone-fixed, and ether-ethanol-fixed cytological specimens. The data indicate that the expression of binding sites with specificity for EPO can be frequently seen in human bronchial malignancies.


Comparative lectinhistochemical studies on paraffin- and glycol methacrylate-embedded CNS tissue specimens from AIDS autopsies. Mistletoe lectin I (ML I) as cell-marker. Brain tissue and spinal cord tissue from 12 patients who had died of AIDS was fixed in neutral formalin; then after the embedment of some of it in paraffin and some of it in glycol methacrylate, it was analyzed lectinhistochemically with mistletoe lectin I (ML I). Mistletoe lectin (ML I) is a reliable marker for microglia cells and macrophages and for special cell forms (polynuclear giant cells, so-called pericytes) belonging to this cell system. In both the embedding procedures used, the representation of the cells is very clear and subtly differentiated so that the preparations are very well suited to the study of AIDS-associated tissue damage in the CNS. Whereas the immunological macrophage markers (CD 68 markers) KPI, PG-M1 produce good results on paraffin-embedded material but are completely useless on material embedded in glycol methacrylate, the cells of the monocyte-macrophage system can be represented very well and reliably after being embedded in glycol methacrylate with mistletoe lectin I (ML I).


[Ultrastructure of amitosis and mitosis of the liver]. Thousands of liver cells of several ultrastructural investigation programs of our laboratory have been evaluated in respect to amitosis and mitosis. Multiplication of hepatocytes by amitosis and mitosis often leads to bi-, tri- and multinucleated cells. No striking ultrastructural changes have been observed in the hepatocytes of human and animal livers. There are no indications of enhanced autophagocytosis or other signs of dedifferentiation. However, non-hepatocytes suffer ultrastructural changes during mitosis: endothelial cells reduce their organelles in number, round up their shape and form microvilli reaching between the microvilli of hepatocytic sinusoidal membrane. Mitotic Kupffer cells contain only small phagosomes and are separated from hepatocytes by endothelial cell processes.


Congenital malformations of the respiratory tract. Malformations of the respiratory tract were found in 21 out of 205 foetuses (10%) in whom various congenital malformations had been prenatally detected by ultrasonography and amniocentesis. Pulmonary hypoplasia was the most common of all pulmonary malformations and was established in 17 cases (81%). Lung alobulation was established in two cases (9.5%), and pulmonary cysts were recorded in another two cases in post-mortem examinations. Correlations established between ultrasonographic and pathomorphological findings were not satisfactory as only 29% of all ultrasonographic diagnosis were complete and correct. Hence, much closer cooperation between the pathologist and clinician is essential to all investigations of congenital malformations, in particular to those of complex nature.


[Syncytial variant of the nodular sclerosing type of Hodgkin's disease in cervical lymph nodes with simultaneous sarcoidosis-like granulomatosis in the intrathoracic lymph nodes and liver]. A woman aged 31 years had been afflicted with mediastinal lymph node enlargement and hepatopathy for two years. Epithelioid-cell granulomatosis was diagnosed at another institute on the basis of biopsies taken from the liver and thoracic lymph nodes, resulting in the differential diagnosis of sarcoidosis or tuberculosis. Another biopsy was taken from enlarged cervical lymph nodes, after tuberculostatic therapy had remained unsuccessful and had not prevented deterioration of the patient's condition. We diagnosed from that biopsy the syncytial variant of nodular sclerosis of Hodgkin's disease. Immunohistochemically, the tumour cells exhibited positive reactions to antigens CD 15 and CD 30, whereas no evidence was provided to the presence of cytokeratins, lysozyme and S-100 protein. In grading, we associated our case with subtype 2 of nodular sclerosis and clinical stage II. Combined radiotherapy and chemotherapy resulted in complete remission of the tumour disease. Presence of granulomatosis similar to sarcoidosis was confirmed by follow-up examination of the liver and lymph node biopsies which originally had been histopathologically examined at another institute. The question is discussed whether or not this granulomatous reaction reflected an increased immunological defence reaction of the organism to Hodgkin's disease and thus offered an explanation for the unexpected favourable course of the patient's disease.


[Aggressive angiomyxoma of the vulva and perineum]. Reported in this paper are two cases of aggressive perineovulvar angiomyxoma. Soft, painless tumours with gelatinous cut surface were detected in either patient, two women aged 32 and 46 years. Histological findings included fibromyxoid stroma with spindle-shaped and stellate cells as well as vascularization in striking abundance. Most of the vessels were thin-walled. There was no plexiform arborization. Strongly atypical nuclei or mitoses or necrotic foci were not recordable. The tumour exhibited infiltrative growth. Tumour cells could be immunohistochemically associated with antibodies against vimentin, whereas negative responses were recorded from antibodies against S-100 protein, factor-VIII-associated protein and pancytokeratin. Aggressive angiomyxoma is a biologically benign neoplasia prone to recurrence and typically localized in soft tissue of the pelvic region. Metastases so far have never been found. Wide local excision has proved to be the optional therapeutic approach.


[Primary intracranial plasma cell granuloma. Report of two cases]. Investigation of biopsies of a suprasellar tumour in a man, 33 years of age, and a supratentorial tumour in a woman, 18 years of age, resulted in histological diagnosis of intracranial plasma cell granuloma, a rare condition. Polyclonality of the plasma cells involved in the process could be verified by immunohistochemical investigation. Chordoma was clinically assumed in the first case and meningioma in the second. Biological behaviour and dubious prognosis are discussed, with reference being made to relevant literature.


Twelve HLA-DR6-related DRB1 alleles and associated DR, DQ haplotypes in traditional Australians and other populations of Asia-Oceania. The relative distributions of 12 HLA-DR6-related HLA-DRB1 alleles in indigenous populations of Australia, Melanesia, Polynesia, Micronesia, and northern and southern China have been determined by analysis of nucleotide sequence polymorphisms in 364 examples of HLA-DR6 positive chromosomes. Oligonucleotide hybridizations of polymerase chain reaction products of HLA-DQA1, DQB1, DRB1 and DRB3 genes generated 24 HLA-DR6-related haplotypes. The study aimed to determine the regional distribution of DR,DQ haplotypes associated with three novel HLA-DR6 alleles, namely DRB1*1408, 1409, and 1410, known to occur in Australian Aborigines, to gain further insights into the molecular phylogeny of these alleles. DRB1*1408 was the most common HLA-DR6 subtype in Oceania, although it was not detected in Chinese. In Australian Aborigines and Papua New Guinean highlanders, DRB1*1408 was associated with DRB3*0202, while in Polynesians and Micronesians it was associated with DRB3*0101. The different haplotype arrangements, together with the near absence of DRB1*1408 in coastal Melanesians, suggest the possibility that two independent mutations have generated DRB1*1408 in Australia and Oceania. DRB1*1409 and 1410 alleles were confined to Australian Aborigines, while DRB1*1407 was found exclusively in Melanesians; DRB1*1401 was the only HLA-DR6 allele represented in all study populations. The population-specific HLA-DR6 alleles and haplotypes have important implications for unrelated bone-marrow donor registries in Australia and Oceania.


Diversity in DNA rearrangements and in RNA expressions of immunoglobulin gene on common variable immunodeficiency. Six heterogeneous common variable immunodeficiency (CVID) patients were analysed for germ-line DNA, DNA rearrangements, and RNA expressions of immunoglobulin (Ig) gene by Southern or northern blotting using appropriate probes. We detected no polymorphism in neutrophil DNA hybridized to a C mu and a C gamma probe. In three patients, both serum Ig and Ig-bearing cells were scarcely detected, and by northern hybridization methods, neither mu mRNA, gamma mRNA, alpha mRNA nor kappa mRNA was detected. However, one Epstein-Barr virus-transformed B lymphoblastoid cell line (LCL) of these three patients was different from the germ line in the region of JH, C gamma, and C kappa, and expressed mu mRNA at a higher level. The B cell defects of these three patients lay on the B cell maturation stage similar to X-linked agammaglobulinaemia (XLA). In two others among the six CVID patients, serum IgM and IgM-bearing cells were detected to a certain degree, and by northern hybridization, mu mRNA was detected at a lower level, but neither mu mRNA, alpha mRNA, nor kappa mRNA was detected. One LCL of these two patients could express mu mRNA at the normal level. In the last patient, the serum IgM was normal, serum IgG and IgA were somewhat low, Ig-bearing cells were normal, mu mRNA and kappa mRNA were detected at the normal level, and gamma mRNA and alpha mRNA were detected at a lower level. The defect of this patient affected the class switch stage. These results showed that primary B cell defects in CVID occurred at several B cell differentiation stages which could be classified by expression of the Ig gene, and at the degree of clonal diversity in the B cell repertoire. Furthermore, this study provides support for the idea that the CVID defect is related to a more generalized cellular function, such as regulating the proliferation and/or clonal expansion of cells of the B lymphoid lineage.


Nucleotide sequencing of HLA-DQ gene second exons in Chinese homozygous cells. Six HLA class I and class II-homozygous Chinese cell lines with unique HLA-Dw types were studied. Since the majority of HLA class II nucleotide sequence polymorphism is localized within the second exons of the genes, we used the polymerase chain reaction (PCR) to amplify these regions in HLA-DQA and DQB genes and subsequently determined the nucleotide sequences. No unique DQA1 or DQB1 alleles were found. However, a new haplotype of DQA1*0601-DQB1*0301-DRB1*1202 was found in two cells; and DQA1*03011 was found in association with DR9 in another two cells. This indicates that new DR-DQ associations may explain the observed new HLA-Dw types. The DQB2 sequences were identical in all six cells and were identical to a sequence previously reported in a DR6 haplotype. The DQA2 sequences from two clones obtained from two cells differed from each other and from previously reported sequences. The results show that the DQA1 and DQB1 alleles in the Chinese individuals studied are as previously reported in Caucasian populations and as such may be typed by restriction fragment-length polymorphism (RFLP) or PCR-sequence-specific oligonucleotide typing (PCR-SSO) or PCR-RFLP using conventional probe or restriction enzyme sets.


Serological and molecular analysis of HLA in Israeli primary sclerosing cholangitis patients. HLA class I and class II were investigated in 15 Israeli primary sclerosing cholangitis patients and compared to healthy controls. None of the well established serological specificities were found to be associated with the disease. HLA-DR52 is serologically defined, but its subtypes DR52a, DR52b, and DR52c cannot be precisely defined by serological means. Therefore, we have used HLA-DNA typing in order to assign the DR52 splits in PSC patients. Genomic DNA was amplified by PCR, dot-blotted and hybridized with sequence specific oligonucleotide probes defining the known HLA-DR52 associated alleles. Only 4 out of the 15 PSC patients tested were found to express DRB3*0101 the allele that encodes DR52a. Of the remaining 11 patients, 9 expressed DRB3*0202 haplotypes, with 2 patients expressing both DRB3*0101 and DRB3*0202, and the remaining 2 patients expressed no DRB3 allele. Our data indicate that there is no apparent association between PSC and the HLA antigens and alleles studied including the alleles of the DRB3 locus in the Israeli population. Thus HLA pheno/genotyping of PSC patients in the Israelis will not be useful for early and/or differential diagnosis of this disease.


A likelihood approach to HLA serology.

A likelihood approach to HLA serology. A likelihood approach to HLA serology has been developed in which the aim is not to define a recognition set for a serum but to describe the serum's ability to react with each and every antigen in the test cells, this ability being quantified in terms of the probability of a positive reaction. For a given set of probabilities, one for each antigen, it is possible to derive the probability of the observed set of reactions (the likelihood of the set of probabilities). The maximum possible value of the likelihood for any possible combination of the probability set can then be sought, but this requires a maximization of likelihood with respect to 60-100 independent parameters. Theoretical considerations of the shape of the likelihood surface prove that, in this particular case, this is a feasible proposition. This approach allows the recognition of three groups of antigens: those for which there is considerable evidence of a specificity, those for which there is either no specificity or a very weak specificity, and those for which there is insufficient evidence on which to base a conclusion. The existence of a specificity can be tested using a log likelihood ratio as a statistic, but the usual assumption of a chi 2 distribution of this statistic cannot automatically be made in this situation. Therefore, the distribution is estimated by simulation. A serologist using this approach would receive considerably more information as to the serum's reaction patterns and valid statistics for the existence, or not, of a specificity.


Frequency of HLA-DPB1 alleles in multiple sclerosis patients from Northern Ireland. Using sequence-specific oligonucleotide probes, no altered distribution in the frequency of HLA-DPB1 alleles was found in multiple sclerosis patients from Northern Ireland. Although present in the controls, linkage disequilibrium between HLA-DPB1*0101 and HLA-DR17 was not found in multiple sclerosis patients.