Analysis of two human monoclonal antibodies against melanoma.

B cells derived from peripheral-blood lymphocytes (PBL) and tumor-infiltrating lymphocytes (TIL) from a patient with a high serum antibody titer to autologous melanoma were transformed with Epstein-Barr virus (EBV) and evaluated for reactivity against autologous tumor. B cells producing antibody reactive with autologous tumor and unreactive with normal fibroblasts were detected both in TIL and in PBL. One cell line derived from PBL and another derived from TIL sustained production of tumor-reactive antibody for 10 weeks and over 15 months respectively. The cell line derived from PBL, 2D11, produced an antibody reactive with a trypsin-resistant antigen expressed on the cell membrane of autologous and allogeneic melanoma cell lines. The cell line derived from TIL, 1F6, produced an antibody reactive with a cell-surface glycoprotein expressed by 5 autologous melanoma cell lines derived from 5 different metastases and 16/19 allogeneic melanoma cell lines. 1F6 also showed reactivity with cell lines derived from a blue nevus, a congenital nevus, an astrocytoma, and 1/4 renal-cell carcinomas; but it was not reactive with 5 foreskin melanocyte cell lines, 2 normal fibroblast lines, 5 leukemia/lymphoma lines, 8 lung-cancer lines, 8 glioblastoma lines, or lines derived from 1 ovarian carcinoma, 1 colon carcinoma, 1 vulvar carcinoma, 1 fibrosarcoma, 1 murine melanoma, or 4 murine leukemia/lymphomas. We describe here an antibody that detects a new melanoma specificity obtained by EBV transformation of tumor-infiltrating B cells.


Cancer-cell traffic in the liver. I. Growth kinetics of cancer cells after portal-vein delivery.

Following the intrasplenic injection of B16F10 melanoma cells into mice, at first single cells, and later multicellular tumor foci were observed at different times in the liver. Cell numbers and tumor volumes were determined over the next 12 days, by confocal microscopy of thick liver sections. Fifteen minutes after injection, approximately 20% of the melanoma cells were identified in the liver microvasculature; after 48 hr, only 0.68% of these retained morphologic integrity; by 5 days only 0.13% of the originally detected cells incorporated BUdR; and, by 12 days, these subsequently grew into tumor nodules. Tumor volume changes with time were not exponential and, following a non-replicative period during the first 2 1/2 days, the volume doubling times increased from approximately 8 to 39 hr at 12 days. BUdR incorporation indicated that this was probably due to a diminishing proliferative fraction in the cancer-cell population. When animals were killed at 12 days, tumor foci were centered on the liver sinusoids and appeared to represent expansive growth of arrested cells, with no evidence of parenchymal invasion by migrating cancer cells, in spite of the absence of a subendothelial basement membrane. Our direct observations indicate that, in this model system, the post-delivery phase of hepatic colonization is characterized by high levels of inefficiency with respect to cancer-cell survival, growth and extravasation.


Proliferation of human colon cancer cells: role of epidermal growth factor and transforming growth factor alpha.

Human colon cancer cells produce and secrete a variety of polypeptide growth factors. The functional role of these growth factors, however, is poorly understood. Though the secretion of epidermal growth factor (EGF)-like activity and EGF-related molecules by human colon cancer cells in culture has been reported, it is not known whether colon cancer cells produce and secrete EGF, and the functional role of EGF in the growth control of these cells is also unknown. We have shown that EGF acts as a potent growth stimulator on the moderately differentiated Moser colon cancer cell line and as an inhibitor on the highly metastatic KM12SM cell line. In the present study, we show that EGF is produced by human colon cancer cells and characterize the levels of EGF mRNA expression and EGF protein secretion from 8 human colon cancer cell lines. The cell-surface EGF receptors on these cell lines were also characterized by radiolabeled ligand binding and Scatchard analyses. All the cell lines expressed EGF mRNA and secreted EGF. Both high- and low-affinity subtypes of EGF receptor were detected on 7 of the cell lines. These lines also secreted transforming growth factor (TGF)alpha. Some cell lines exhibited a proliferative response to treatment with either exogenous EGF or TGF alpha, while others did not respond to treatment with these growth factors. Antibody-blocking experiments, using anti-EGF or anti-EGF receptor antibody, suggested that these cell lines could be broadly classified into 2 groups in terms of their autocrine or paracrine growth regulation via the cell-surface EGF receptor: (1) cells that utilized EGF and/or TGF alpha; and (2) cells that did not utilize EGF or TGF alpha (via the cell-surface receptor), even though they secreted abundant amounts of these growth factors.


Cholesterol consensus: a trans-Atlantic perspective. Amsterdam, The Netherlands, 31-October-1 November 1991.

This consensus document has been drawn up following the round-table conference 'Cholesterol Consensus: a Trans-Atlantic Perspective', Amsterdam, 31 October and 1 November 1991. Its recommendations are as follows. Elevated plasma cholesterol level should be viewed as one of a number of important risk factors that contribute to the high incidence of coronary heart disease in industrialized societies. Control of lipid abnormalities forms an essential part of an overall preventive strategy of risk factor reduction. Both population and individual strategies are required. To reduce the mean plasma cholesterol levels of populations, a multidisciplinary approach is needed, involving the health-care professions, government, food manufacturers and distributors, community and consumer organizations, and the media. While it is desirable for all individuals to know their plasma cholesterol level, universal screening is recommended only where resources for treatment and follow-up are sufficient to cope with the case load detected. As an interim measure, screening should be focused on individuals at special risk. The recommendations of the European Atherosclerosis Society and the U.S. National Cholesterol Education Program should be followed with regard to thresholds for treatment by dietary counselling and pharmacological methods, and targets for cholesterol reduction. These recommendations should be periodically reviewed as new research data becomes available. A number of key areas for research are listed.


Catecholamine excretion and heart rate as factors of psychophysical stress in table tennis.

Table tennis, like tennis, squash and badminton, is a racket sport. All these sports have in common a rapid succession of mostly short-term maximal or submaximal efforts and short recovery phases. The goal of this paper is to investigate the psychophysical stress in table tennis by means of the stress hormones epinephrine and norepinephrine. The catecholamines were determined from urine samples. 16 Austrian top-level table tennis players were examined. There were 8 female and 8 male players in this group. The catecholamine excretion at rest (R), training (TR), practice competition (PC), competition (C) and treadmill ergometry (TE) are indicated in ng/min of collecting time. When the group is divided according to sex, we find marked differences in the catecholamine release. While the epinephrine excretion during and after training and practice competition is basically the same, it is lower during and after treadmill ergometry and higher after competition. The same result was found with respect to norepinephrine excretion. The ratio between norepinephrine and epinephrine was 4:1 at rest and during and after treadmill ergometry, 6:1 during and after training, 5:1 during and after the practice competition and 2:1 during and after the competition. The investigation showed that a table tennis competition puts high stress on the player. The mental component should therefore receive much more importance in order to keep the stress low.


Haemorheology and exercise: intrinsic flow properties of blood in marathon running.

Haemorheological and haematological parameters were studied in venous blood samples taken from 8 endurance-trained athletes before and after a marathon run. Viscometry was performed in a 20 microns glass capillary and in a Couette viscometer. Apparent blood viscosity was lower in the capillary than in the rotational viscometer, but significant differences between pre- and post-run samples were not observed in either system. This is presumably due to the absence of changes of haematocrit (pre-run [mean +/- SD]: 0.453 +/- 0.016, post-run: 0.456 +/- 0.018). Although protein concentration increased significantly from 70.8 +/- 2.1 g/l (pre-run) to 76.0 +/- 3.9 g/l (post-run), the difference of plasma viscosity was not statistically significant (pre-run: 1.26 +/- 0.03 cP, post-run: 1.30 +/- 0.07 cP). In all samples, plasma viscosity showed a significant positive correlation with total protein, globulin, albumin, but not with fibrinogen concentration. Cone-plate aggregometry and a filtration technique were used to evaluate red cell aggregation and deformability, two determinants of blood rheology which also were not different between pre- and post-run samples. The absence of viscosity changes in relation to the marathon run may be due to the relative stability of blood and plasma volume, which can be deduced from haematological parameters. Alterations of plasma composition (e.g. acidosis or hyperosmolality) seem to cause only minor, if any, changes of microrheological cell properties.


Determining the variability of performance on Wingate anaerobic tests in children aged 6-12 years.

In two parts, this study investigated variability of performance in Wingate anaerobic tests (WAnTs) of normal, healthy, male children aged six, eight, ten and twelve years. The purpose of the first part of the study was to investigate the mean coefficient of variation (C.V. = [S.D./X].100) of children who performed four WAnTs over a period of four weeks. The results revealed age-related differences in the performance data of peak power (PP) and mean power (MP) but no differences between age groups in fatigue index (FI%) = [PP-Pend)/PP.100]. C.V. data from this group of children did not differ significantly between ages in any of the dependent variables. Coggan and Costill (5) measured the variability of male adults during 30 seconds of highly intense cycling and reported C.V.s in PP and MP performances of 6.7 and 6.5%, respectively. C.V.s of children in the first part of the present study were 7.3 and 6.8% for PP and MP, respectively. The C.V. in the FI% of the children (26.7%) however differed substantially from the previously studied adults (10.3%). The second part of the study was conducted to examine the C.V. in FI% performances of children in the WAnT. The purpose of this part of the study was to examine the extent to which a computerized onscreen game, which was linked to pedal cadence affected the variability of children performing WAnTs. A significant decrease was obtained in the C.V. of the FI% in the presence of the computerized feedback game (16.3 and 23.6, with and without the game, respectively). It is suggested that game-based testing procedures may ensure more consistent result in the assessment of pediatric populations.


A new approach for the determination of ventilatory and lactate thresholds.

In order to determine the ventilatory threshold (VT) and the lactate threshold (LT) in a reliable way, a new method is proposed and compared with conventional methods. The new method consists of calculating the point that yields the maximal distance from a curve representing ventilatory and metabolic variables as a function of oxygen uptake (VO2) to the line formed by the two end points of the curve (Dmax method). Male cyclists (n = 8) performed two incremental exercise tests a week apart. Ventilatory/metabolic variables were measured and blood was sampled for later lactate measurement during each workload and immediately after exercise. No statistical differences were observed in the threshold values (expressed as absolute oxygen uptake; VO2) determined by the Dmax method and the conventional linear regression method (according to O2 equivalent; EqO2) and venous blood at the onset of blood lactate (OBLA), while VT assessed with the conventional linear method (according to the slope of CO2 output; Vslope) yielded significantly lower threshold values. Similar results were obtained from the reproducibility test. Thus, the Dmax method appears to be an objective and reliable method for threshold determination, which can be applied to various ventilatory or metabolic variables yet yield similar results. The results also showed that breathing frequency can be used to determine VT.


Accuracy of pulse oximetry during exercise stress testing.

Pulse oximetry is used extensively during exercise stress-testing in the clinical and sports medicine settings. There are few validation studies to assess the appropriateness of using pulse oximetry under conditions of potentially compromised peripheral blood flow. To study the accuracy of pulse oximetry during severe exercise stress, 10 athletes undertook 3 bouts of exhaustive exercise; once at an intensity requiring VO2max (max), once at 115% of VO2max (Smax), and once at Smax while FIO2 was increased to 0.30. The results indicate relatively large underestimations occur when pulse oximetry is used to estimate %SaO2 during exercise, when compared to the criterion samples of gas analysis in arterial blood. These differences were exacerbated as the exercise intensity increased from a mean(+/- SE) difference of 2.9 +/- 0.7 %SaO2 at max to 4.6 +/- 0.7 %SaO2 at Smax. Breathing a higher FIO2 reversed the hypoxemia that occurred during the normoxic exercise, however, pulse oximetry measurements failed to detect this alteration in %SaO2. Estimates of oxygen saturation during severe exercise using pulse oximetry should be viewed with caution, as potentially large errors may occur.


Maximal-intensity intermittent exercise: effect of recovery duration.

Seven male subjects performed 15 x 40m sprints, on three occasions, with rest periods of either 120 s (R120), 60 s (R60) or 30 s (R30) between each sprint. Sprint times were recorded with four photo cells placed at 0, 15, 30 and 40 m. The performance data indicated that whereas running speed over the last 10 m of each sprint decreased in all three protocols (after 11 sprints in R120, 7 sprints in R60 and 3 sprints in R30), performance during the initial acceleration period from 0-15 m was only affected with the shortest rest periods increasing from (mean +/- SEM) 2.58 +/- .03 (sprint 1) to 2.78 +/- .04 s (spring 15) (p < .05). Post-exercise blood lactate concentration was not significantly different in R120 (12.1 +/- 1.3 mmol.l-1) and R60 (13.9 +/- 1.2 mmol.l-1), but a higher concentration was found in R30 (17.2 +/- .7 mmol.l-1) (p < .05). After 6 sprints there was no significant difference in blood lactate concentration with the different recovery durations, however, there were significant differences in sprint times at this point, suggesting that blood lactate is a poor predictor of performance during this type of exercise. Although the work bouts could be classified primarily as anaerobic exercise, oxygen uptake measured during rest periods increased to 52, 57 and 66% of maximum oxygen uptake in R120, R60 and R30, respectively. Evidence of adenine nucleotide degradation was provided by plasma hypoxanthine and uric acid concentrations elevated post-exercise in all three protocols. Post-exercise uric acid concentration was not significantly affected by recovery duration.(ABSTRACT TRUNCATED AT 250 WORDS)


Physiological changes in male competitive cyclists after two weeks of intensified training.

To study the physiological response to heavy training, seven male competitive cyclists intensified their normal training program for two weeks (IIT) in order to achieve a state of short-term overtraining. The subjects underwent a graded cycle ergometer test to exhaustion, an outdoor 8.5 km time trial and a computerized test to study reaction time and visual perception, before, during and after the two weeks of intensified training and after two weeks of recovery. Furthermore subjects kept a daily log in the form of a questionnaire. After two weeks of IIT all subjects showed symptoms of overtraining: the general state of well being declined as indicated by the questionnaire while performances on time trial (mean +/- SEM: 830 +/- 14 sec-871 +/- 19 sec), contests and maximal power output (mean +/- SEM: 336 7 watt-310 +/- 5 watt) declined significantly. Maximal (mean +/- SEM 11.8 +/- 1.1 mmol.l-1-5.9 +/- 0.5 mmol.l-1) and submaximal lactate values were significantly lowered during ergometer test after the IIT, while the workload at the 4 mmol point increased significantly (mean +/- SEM 234 +/- 10 watt-267 +/- 13 watt). Sleeping heart rate increased significantly (mean +/- SEM 49.5 +/- 9.3 BPM-54.3 +/- 8.8 BPM). Maximal heart rate (mean +/- SEM 185 +/- 3 BPM-178 +/- 2 BPM, mean heart rate during the time trial (mean +/- SEM 178 +/- 2 BPM-169 +/- 2 BPM) and VO2max (mean +/- SEM 4801 +/- 121 ml.min-1-4409 +/- 101 ml.min-1) were all significantly lowered by the IIT.(ABSTRACT TRUNCATED AT 250 WORDS)


Response of beta-endorphin and estradiol to resistance exercise in females during energy balance and energy restriction.

The acute effect of weightlifting on beta-endorphin and estradiol was studied in experienced female recreational weightlifters. Five eumenorrheic females completed two months of testing, each with a different sequence of testing conditions (SEQ1 and SEQ2). In SEQ1, a week of weight maintenance diet and prescribed exercise (3 d.wk-1, 3 sets, approximately 85% 1 RM, 10-12 reps, eight lifts) beginning on d 11 of their menstrual cycle was followed by measurement of hormone response to a weightlifting bout during energy balance (EBAL) on d 18. This included blood sampling via a catheter before, just after, and at 15 and 30 min of recovery. The women consumed 500 kcal per day for the next 48 hrs and then repeated the weightlifting test during negative energy balance (NEBAL). SEQ2 was similar except that the 48 hrs of NEBAL preceded the EBAL test condition. Estradiol and beta-endorphin increased from baseline to immediately post exercise under both dietary conditions but was significant only during NEBAL. Estradiol increased 1.6 fold and beta-endorphin 3.7 fold by the end of the resistance exercise bout during NEBAL. Both hormones were also elevated for a longer time during recovery in the NEBAL condition. Since estradiol and beta-endorphin can suppress gonadotropin release, it is possible that repeated elevations in these hormones during weightlifting, especially concurrent with energy restriction, could contribute to disruption of the menstrual cycle.


Seasonal deterioration of selected physiological variables in elite male skiers.

The purpose of this study was to determine whether seasonal deterioration in physiological variables could be observed in skiers. Eighteen international male British downhill, free-style, and speed skiers were subjected to a maximal treadmill running test, a 30-s Wingate test, and isokinetic dynamometry at the beginning, middle, and end of the 1989-90 competition season. Maximal oxygen intake (VO2max) and respiratory anaerobic threshold (T vent) were among the parameters measured on the treadmill test, while peak and mean power outputs were measured during the Wingate test. During dynamometry, knee flexors and extensors were bilaterally assessed at 1.04 and 3.14 rad.s-1. Mean VO2max (p < 0.01) and mean T vent (p < 0.05) were lower at the end compared to the beginning, but not compared to the middle of the competition season. The isokinetic test demonstrated lower mean peak torques at 1.04 rad.s-1, for the knee extensors measured at the end of the season, compared with both the start (p < 0.01) and the middle (p < 0.05). Also at 1.04 rad.s-1, knee flexors produced lower torques at the end than the start of the season (p < 0.05). No further statistical differences were found. It was concluded that seasonal deterioration in key physiological variables such as aerobic endurance and muscle strength, can be observed in elite alpine skiers, and that in-season fitness training programmes should take account of this.


The effect of exercise on anterior knee laxity in female basketball players.

To study the effect of physical activities on the anterior laxity of the knee joint, the anterior knee laxity of female semi-professional basketball players was measured during a typical day. The participants worked in the office in the morning, and practiced in the afternoon, which included a 60 minute warm-up, followed by a 150 minute game style practice. The laxity was measured six times in one day at different activity levels. The anterior knee laxity did not change during sedentary work in the morning, but increased significantly with game style practice. After 1 1/2 hours the increased anterior knee laxity had not recovered completely. However, after five hours it had recovered completely.


Vitiligo in children.

In our study the relative incidence of vitiligo among new patients was 2.6%. Twenty percent were children and 74% were adults. Of the 90 children, 38.9% were boys, and 61.1% were girls. This sex difference was statistically highly significant. The adult sex-ratio was not statistically significant. The relative incidence of the clinical subtypes in children and adults was compared, and the difference was found to be statistically highly significant only in the case of vitiligo vulgaris and segmental vitiligo. On the basis of the difference in the sex-ratio and in the relative incidence of the subtypes of vitiligo vulgaris and segmental vitiligo, we feel that childhood vitiligo is a distinct subtype of vitiligo.


Specific cutaneous lesions in a CD8+ peripheral T-cell lymphoma.

Histopathologic, immunohistochemical, and ultrastructural studies were carried out on cutaneous lesions of a 43-year-old man with an aggressive peripheral T-cell lymphoma involving the lung, central nervous system, bone marrow, and skin. Some results are distinctive and not previously reported, such as extremely strong epidermotropism, aberrant CD8+ immunophenotype with lack of one pan T antigen (CD5), and giant cytoplasmic granules. We discuss these features comparing them with other hematologic malignancies usually involving the skin, such as cutaneous T-cell lymphoma, adult T-cell leukemia/lymphoma, angiocentric lymphomas, and malignant histiocytosis.


Lack of predictive factors in late recurrence of stage I melanoma.

Through the UCLA tumor registry, patients treated for late-relapse malignant melanoma (after a 10-year disease-free interval) were identified and studied retrospectively. The patients were studied in regards to age, sex, primary site of the lesion, the disease-free interval, elective lymph node dissection, mode of recurrence and survival time following relapse. The histologic characteristics studied included the maximal tumor thickness, Clark's levels, rate of mitoses, and presence of ulceration. Based on the 0.84% incidence of late-relapse, the recommendation for annual life-time follow-up examinations for all patients with malignant melanoma is supported.


Reaction in leprosy: acute phase reactant response during and after remission.

Sera from 25 patients with type 1 (Lepra), upgrading and downgrading, and type 2 (erythema nodosum leprosum [ENL]) reactions were assayed, during the reaction and after its clinical remission, for changes in levels of alpha-1-antitrypsin (A1A) and C-reactive protein (CRP). The results were compared with those from normal healthy adults and patients of leprosy without history and/or clinical evidence of reaction. The A1A levels correlated better with changes in status of type 1 reaction; whereas CRP levels correlated well with alterations in type 2 reactions and were definitely superior to A1A in this situation for monitoring the course of these episodes.


Multiple basal cell carcinoma in tropical Australia.

No association between HLA DR1 and the development of multiple basal cell carcinomas (BCC) was found among patients who had lived at least two-thirds of their lives in the tropics. The percentage of patients with multiple BCCs increased with age; this was different from what has been found in people living in the temperate zone of Australia.


Cutaneous myiasis: a simple and effective technique for extraction of Dermatobia hominis larvae.

We describe two patients with cutaneous myiasis caused by Dermatobia hominis. Three larvae were removed successfully by a simple technique that involved the injection of each larva with 2 mL of lidocaine, making surgical extraction by incision and exploration unnecessary.


Lymphedema treated by microwave and elastic dressing.

For lymphedema, surgical treatment is contraindicated and other conservative treatments including oral medicine are not sufficient. Treatment with microwave and elastic dressing has been evaluated in 30 cases of lymphedema with 11 control patients who received only one of the two treatments. Twenty-four patients were effectively treated. In six, edema was not decreased as compared with reduction due to daily physical activity. It was observed that the earlier the patient was treated after onset of edema, the better the results. Six of 10 control patients who received only elastic dressing showed slight decrease in edema. A patient who received only microwave did not show any improvement. There was no recurrence in half of the treated patients. Treatment of lymphedema by microwave and elastic dressing appears satisfactory, particularly when given at an early stage of the disease.


The survival of Ostertagia circumcincta and Trichostrongylus colubriformis in faecal culture as a source of bias in apportioning egg counts to worm species.

When cultured alone or concurrently with Trichostrongylus colubriformis in sheep faeces, Ostertagia circumcincta produced fewer infective larvae per 100 eggs than did T. colubriformis. Averaged over five trials 60% of T. colubriformis eggs were recovered as infective larvae while for O. circumcincta the figure was only 39%. This result was observed for two strains of O. circumcincta and was independent of when larvae were harvested from culture (days 6-10 at 25 degrees C). The mortalities of both species occurred at the first and second larval stages. These observations are of concern when using larval differentiation from faecal culture to make quantitative estimates of worm egg numbers for each species present. Species such as T. colubriformis which have a low mortality during culture are likely to have their egg numbers overestimated when cultured with a species, like O. circumcincta, that suffers high mortality in culture.


Metabolism of lipid peroxidation products by the gastro-intestinal nematodes Necator americanus, Ancylostoma ceylanicum and Heligmosomoides polygyrus.

Somatic extracts of the three parasitic nematodes Necator americanus, Ancylostoma ceylanicum and Heligmosomoides polygyrus were able to detoxify a model hydroperoxide and a putative natural peroxide by glutathione-dependent peroxidase activity while cytotoxic carbonyls could be metabolized by NADPH-linked reduction activities. Unlike cestodes and digeneans, the nematodes in this study could not enzymatically conjugate carbonyls with glutathione. The results indicate that the three nematodes can protect themselves against possible host-immune initiated lipid peroxidation of their membranes at the level of the hydroperoxide and at the level of cytotoxic carbonyl, although other protective enzymatic mechanisms are also likely to exist (superoxide dismutase and catalase).


A strategy for production of monoclonal antibodies to Echinococcus granulosus antigen 5 and antigen B.

Serum antibody responses to sheep hydatid cyst fluid (SHCF) and a purified Antigen 5 (Ag5) were examined in ELISA, immunoelectrophoresis (IEP) and immunoprecipitation (IP) to facilitate production of monoclonal antibodies (MAb) to E. granulosus Ag5 and Antigen B (AgB). Although sera from mice immunized with SHCF contained antibodies of various classes, the fusions using these donor mice resulted in mainly anti-AgB MAb, possibly due to the preferential selection of MAb to AgB by the SHCF-based ELISA screening system. Donor mice immunized with Ag5 also produced several classes of antibodies, and the resultant fusions enabled selection of IgG MAb to Ag5.


Echinococcus granulosus in sheep: transfer from ewe to lamb of 'Arc 5' antibodies and oncosphere-killing activity, but not protection.

Fourteen ewes were orally dosed with 2000 E. granulosus eggs at 2 weeks of age, were mated at 19 months, and produced lambs when the cysts were 2 years old. One week after parturition, all 14 ewes had 'Arc 5' antibodies in their serum, as did 11/14 of their lambs. Fourteen uninfected ewes were immunized three times before parturition with E. granulosus eggs injected intramuscularly. Cysts grew at the first, or first and second site, but not the third, indicating that the ewes were immune prior to parturition. Most sera from these ewes and their lambs, and from 14 control ewes and their lambs, produced precipitin arcs with hydatid cyst fluid, but no 'Arc 5'. All lambs were challenged with 500 eggs 2 weeks after birth. At necropsy, cyst numbers within groups ranged from 3 to > 200, but there was no significant difference between the three groups of lambs. The immunized ewes did not pass a protection to their lambs that was effective when the lambs were challenged. 'Arc 5' antibodies were induced by prolonged infection with cysts, and were not seen in the sera of ewes immunized with eggs, although the eggs developed into cysts at the injection sites. 'Arc 5' antibodies did not protect lambs against infection, and were not correlated with protection in ewes. Subcutaneous injection of oncospheres into four ewes from each group at the time of lamb challenge showed that the immunized ewes were immune to this method of challenge, but the infected and control ewes were not.(ABSTRACT TRUNCATED AT 250 WORDS)


Apparent lack of genetic variation within Pelecitus roemeri (Nematoda: Filarioidea) from three Australian species of macropodid marsupial.

An electrophoretic study of Pelecitus roemeri from Macropus robustus, M. giganteus and Wallabia bicolor revealed no genetic differences at 23 enzyme loci. The genetic data support the existing morphological evidence that P. roemeri from these three hosts represents a single species. The data show no genetic variation between nematodes from the same or different host species collected in northern and southern Australia. This result is discussed briefly in relation to Price's model of parasite speciation.


Mucosal mast cell response to Hymenolepis diminuta infection in different rat strains.

Five-week-old DA male rats infected with 10 Hymenolepis diminuta cysticercoids showed significant mastocytosis 6 weeks post-infection and low persistence of worms. In F344/N rats, however, no mastocytosis and no worm loss occurred during a 6 week infection. Mucosal mast cells appear to be associated with the expulsion of H. diminuta from DA rats.


Persistence of immune response in human toxocariasis as measured by ELISA.

The antibody titer was followed in a group of patients, clinically diagnosed with toxocariasis, during a 5 year period. We observed that larvae can survive for at least 5 years in humans. Antigenic stimulation was enough to keep high levels of immunoglobulins over this period. Antibody levels decreased slowly and this pattern is similar to that shown by animal models.


Sensitive detection of trypanosomes in tsetse flies by DNA amplification.

African trypanosome species were identified using the Polymerase Chain Reaction (PCR) by targeting repetitive DNA for amplification. Using oligonucleotide primers designed to anneal specifically to the satellite DNA monomer of each species/subgroup, we were able to accurately identify Trypanosoma simiae, three subgroups of T. congolense, T. brucei and T. vivax. The assay was sensitive and specific, detecting one trypanosome unequivocally and showing no reaction with non-target trypanosome DNA or a huge excess of host DNA. The assay was used to identify developmental stage trypanosomes in the tsetse fly. The use of radioisotopes was not necessary and mixed infections could be detected easily by incorporating more than one set of primers in a single reaction. The use of crude preparations of template made the process very rapid. The methodology should be suitable for large-scale epidemiological studies.


Effect of genetic type, lactation and management on helminth infection of ewes in an intensive grazing system on irrigated pasture.

A survey of helminth infection was conducted in a flock of 290 ewes distributed into Romanov (R), Merinos d'Arles (M) and Romanov x Merinos (R x M) genetic types, grazing irrigated pasture in the south of France. Faecal egg and larval counts were done seven times per year from 1981 to 1984 on homogeneous groups of ewes and then individually once to four times every autumn from 1985 to 1988. Helminth fauna was diverse and more abundant during autumn. High levels of strongyle infection occurred in the ewes that remained on the same irrigated pastures during summer. Moving to Alpian pastures during the summer lowered autumnal infection. Significant differences between genotypes in intensity of infection were observed in the order R > R x M > M for strongyles (Teladorsagia circumcincta and/or Trichostrongylus vitrinus, Chabertia ovina and/or Oesophagostomum venulosum, Nematodirus spp.), Moniezia spp. and Dictyocaulus filaria. The hierarchy was reversed for Fasciola hepatica infection and not consistent from one year to another for protostrongylid infections. The effect of lactation intensity on the postparturient rise was studied by equilibrating number of ewes according to reproductive status (zero, one or two lambs in lactation). Merino ewes with two lambs in lactation, as well as primiparous Romanov ewes, had significantly higher strongyle infections than the others. The repeatabilities of the larval and egg counts between the four trial years were 0.24, 0.23 and 0.16, respectively, for protostrongyles, Nematodirus and strongyles, with higher intra-annual values for protostrongyles and inconsistently significant results for strongyles due to the presence of several species.


Synergistic growth studies of Entamoeba gingivalis using an Ecologen.

A unique multiple diffusion growth chamber, an Ecologen, designed for the study of interactions among microorganisms, was introduced as a means of growing xenic cultures of Entamoeba gingivalis with Crithidia sp. or Yersinia enterocolitica. Entamoeba gingivalis was grown in the central diffusion reservoir of the Ecologen connected to separate growth chambers inoculated with the microorganisms to be evaluated. Growth of the accompanying bacteria in the E. gingivalis compartment was almost completely eliminated, except for sparse Pseudomonas sp. growth. The most vital E. gingivalis cultures were observed when either Crithidia sp. or Y. enterocolitica were added to the Ecologen 48 h prior to the E. gingivalis inoculum. The medium which provided the best growth of the oral protozoan in this system was the new improved E. gingivalis medium containing antibiotics.


Dictyocaulus viviparus: isolation and characterization of a recombinant antigen with potential for immunodiagnosis.

Crude adult worm antigen of Dictyocaulus viviparus was examined for specific antigens by SDS-PAGE and immunoblotting using sera from cattle experimentally infected with D. viviparus, vaccinated with a normal or a reduced dosage of the commercial lungworm vaccine, and helminth-free cattle. A D. viviparus-specific region M(r) 18,000 was identified and isolated. A lambda ZAP II cDNA expression library consisting of 4.4 x 10(5) recombinant clones (88% of the total number of clones) was constructed from D. viviparus adult worm mRNA. Rabbit antiserum to the M(r) 18,000 antigen was used to screen the cDNA library and eight positive clones were picked and allocated to the same antigenic family by sibling analysis. All clones were subcloned into the plasmid pGEX-2T, and the clone with highest expression yields was expressed as a glutathione S-transferase fusion protein (DvGST3-14) or, after cleavage with thrombin, as pure recombinant parasite protein (Dv3-14). The native parasite antigen encoded by the clone was identified. The immunodiagnostic potential of the recombinant proteins was assessed by immunoblotting.


The kinetics of mebendazole binding to Haemonchus contortus tubulin.

The kinetics of the binding of mebendazole (MBZ) to tubulin from the third-stage (L3) larvae of the parasitic nematode, Haemonchus contortus, have been characterized. In partially purified preparations, the association of [3H]MBZ to nematode tubulin was rapid, k1 = (2.6 +/- 0.3) x 10(5) M-1 min-1, but dissociation was slow, k-1 = (1.58 +/- 0.02) x 10(-3) min-1. The affinity constant (K(a)) for the interaction, determined by the ratio k1/k-1, was (1.6 +/- 0.2) x 10(8) M-1. Similar results were obtained with crude cytosolic fractions. In equilibrium studies, performed with partially purified nematode tubulin under similar conditions, a K(a) of (5.3 +/- 1.6) x 10(6) M-1 was obtained. The best estimate for the K(a) of the MBZ-nematode tubulin interaction is considered to be the 'kinetic' value determined from the ratio of rate constants. The slow dissociation of MBZ from nematode tubulin, which contrasts with the rapid dissociation of MBZ from mammalian tubulin, supports the hypothesis that the selective toxicity of the benzimidazole anthelmintics results from a difference between the affinities of mammalian and nematode tubulins for these drugs.


Selection of Merino sheep for increased and decreased resistance to Haemonchus contortus: peri-parturient effects on faecal egg counts.

Peri-parturient worm egg counts were compared in 395 Merino ewes bred for either increased (IRH) or decreased (DRH) resistance to Haemonchus contortus, or from an unselected control flock (CH). Following a 10 month period with no anthelmintic treatment, a rise in egg counts began about 4 weeks before parturition and continued into lactation. At all stages, egg counts were significantly lower in IRH ewes, even those failing to conceive. Before lambing, IRH ewes averaged 263 epg, compared with 1113 epg in CH and 1618 epg in DRH ewes. After 2 weeks of lambing, lactating ewes averaged 1050, 2173 and 3708 epg, respectively, in the three lines. At the end of lambing, egg counts had increased to 1645, 3959 and 4124 epg, respectively. Counts appeared to have peaked in IRH ewes but not in DRH ewes. Ewes with twins had higher counts than those with singles, and ewes suckling lambs had higher counts than those losing lambs. No significant variation was associated with age of ewe (2-7 years). Although there were several nematode species present, the results suggest that, in comparison with DRH ewes, IRH ewes were particularly effective in reducing pasture contamination with H. contortus, both before and during the peri-parturient period.


Association of blood eosinophilia with the expression of resistance in Romney lambs to nematodes.

The dynamics of blood eosinophilia was studied in Romney lambs dosed twice weekly with 5000 infective larvae of Trichostrongylus colubriformis. A marked rise in blood eosinophil counts of the dosed lambs coincided with the start of the decline of faecal egg counts (FEC). Eosinophilia was also studied in grazing Romney lambs from a breeding programme, based on selection for high or low FEC. Comparison of the sire groups suggested that the magnitude of the eosinophil response was under genetic control. The significant negative correlations between blood eosinophil counts and FECs confirmed the association between eosinophilia and the expression of resistance in Romney lambs to nematodes. However, comparison of the mean blood eosinophil counts and FECs for both the ram and ewe lambs of the resistance sire groups suggested that the association of eosinophilia and resistance may be greater for some sire groups than others.


Inflammatory responses in the intestine during tapeworm infections. Mucosal mast cells and mucosal mast cell proteases in Sprague-Dawley rats infected with Hymenolepis diminuta.

Comparative studies were made of two populations of Sprague-Dawley rats infected with Hymenolepis diminuta. The time course of infection, the development of mucosal mastocytosis and the levels of rat mucosal mast cell (MMC) protease (RMCP II) in serum and in jejunal mucosal tissues were monitored at intervals after infection with 40 cysticercoids of the tapeworm. Worm expulsion patterns differed markedly between the two populations, rats of New Zealand origin showing an abrupt and clear-cut loss of worms, rats of English origin showing a more gradual decline over a longer time period. In both populations, however, numbers of MMC and levels of tissue RMCP II were positively correlated with time after infection and negatively correlated with worm numbers. In only one of the three experiments (using English strain rats over a short time period) did levels of serum RMCP II change with time. In the other two experiments, in which English-strain and New Zealand-strain rats were used, there were no correlations between serum RMCP II and time, numbers of MMC, numbers of worms or levels of tissue RMCP II. The absence of correlation between serum RMCP II and worm loss in these experiments implies that MMC have no direct role in expulsion of H. diminuta. The data do show, nevertheless, that this purely luminal tapeworm is fully capable of activating the mucosal T lymphocyte-MMC precursor axis to elicit a mucosal mastocytosis.


Secondary Echinococcus multilocularis infection in severe combined immunodeficient (scid) mice: biphasic growth of the larval cyst mass.

E. multilocularis infection was suppressed in C.B-17 mice after intraperitoneal inoculation of protoscoleces, with larval cysts weighing no more than 1.0 g. In scid mice, which are genetically identical to C.B-17 except for a deficiency in functional lymphocytes, infection progressed and larval cysts reached a mass of 17.5 g at 15 weeks post-infection. The growth of the larval cyst mass in scid mice was similar to that in other susceptible mouse strains, with a biphasic pattern. Histological observations revealed giant cells and granulomatous inflammation in the C.B-17, but not in the scid mice. These results led to the conclusion that suppression of the growth of the larval cyst mass in the initial stage of infection in susceptible mice strains is caused by factors other than the host's lymphocytic immune response.


The effect of adjuvant and specific or non-specific vaccination on development of protective immunity of rabbits against Trichostrongylus colubriformis infection.

Adult New Zealand rabbits were vaccinated subcutaneously with one dose of 100 micrograms adult nematode phosphate buffered saline-soluble proteins (PBS-ASP, groups I and II), a detergent-soluble fraction of adult somatic proteins (DS-ASP, group III) or three doses of 1 mg normal rabbit serum proteins (group IV). Injections of the immunogens in groups II, III and IV were accompanied with beryllium hydroxide, Be(OH)2 as an adjuvant. Vaccinated rabbits and also those of group V (naive) were challenged orally with 10,000 infective larvae of T. colubriformis 14 days after antigen injection and necropsied 2 weeks later. A single dose of PBS-ASP induced 33.5% protection when the antigen was given alone (group I) and 69.4% when injected with Be(OH)2 (group II). A detergent-soluble fraction of ASP given with the adjuvant provided 87.2% protection (group III), whilst non-specific vaccination with serum proteins plus Be(OH)2 elicited 99% protection (group IV). Mesenteric lymph node leukocyte responses were measured using a leukocyte migration inhibition assay. A significant response was observed only in group IV. In ELISA tests IgA antibodies specific to PBS-ASP reached the highest level in the intestinal mucosa of groups I and II and in the bile of groups I and III. Antibody levels of IgG isotype were similar in the intestinal mucosa of all the immunized groups. Nematode antigen was detected using a 'sandwich' ELISA method in faecal protein extracts of rabbits of groups II and III on days 2-6 after challenge.


Population dynamics of Trichostrongylus colubriformis and Ostertagia circumcincta in single and concurrent infections.

Twenty-one-week-old worm-free pen-reared lambs were infected weekly with either 10,000 T. colubriformis larvae, 5000 O. circumcincta larvae, or with both species (15,000 larvae per week). Larval establishment and total worm burdens were estimated after 4, 7, 10 and 13 weeks of infection. Faecal egg counts and lamb bodyweights were measured weekly, and numbers of eosinophils in blood were estimated before infection and at weeks 5, 8 and 14. For both species of worms, the dynamics of infection (establishment, worm burdens, egg counts) were not affected by concurrent or pre-existing infection with the other species. Infection with T. colubriformis alone did not protect against O. circumcincta, but infection with O. circumcincta alone provided slight protection against the T. colubriformis larvae. Blood eosinophils increased between 5 and 8 weeks of infection and were similar for the three infections. This corresponded to the reduction in establishment for both species.


Immunoperoxidase labelling of albumin at the endothelial cell surface of frog mesenteric microvessels.

Albumin was visualised at the endothelial cell surface of perfused frog mesenteric microvessels using immuno-peroxidase labelling. Vessels in the mesenteries of pithed frogs were washed free of blood and then perfused with frog Ringer solutions containing bovine serum albumin (BSA) at a concentration of 20 mg BSA ml-1, followed by a brief Ringer flush to remove excess albumin from the vessel lumen. The tissues were fixed in 1% glutaraldehyde and a double antibody labelling technique used to identify albumin within the tissue. A dense layer of peroxidase reaction product was seen, which extended 25-50 nm into the vessel lumen. It appeared as a continuous layer lining the luminal openings of interendothelial cell clefts and vesicles open to the luminal cell surface. In some vessels a more irregular layer of peroxidase labelled albumin was seen extending 150 to 200 nm into the vessel lumen, whilst in others clumps of peroxidase labelled albumin were also seen within the vessel lumen. These data offer direct evidence that BSA does interact with the endothelial cell surface of perfused frog mesenteric microvessels but suggest that a proportion is loosley or non-specifically bound to the cell surface and can be removed by a brief Ringer flush. The remainder appears more tightly bound and resistant to Ringer flush.


Subcutaneous interstitial fluid pressure and arm volume in lymphoedema.

Interstitial fluid pressures were measured by the wick in needle method in the swollen and normal arms of 38 patients with lymphoedema resulting from treatment for breast cancer. The mean increase in arm volume, calculated from sequential circumferential measurements, was 33% (range 0.25 to 85.9). Subcutis interstitial fluid pressure in the swollen arm (2.0 cmH2O range -4.5 cmH2O to 6.8 cmH2O) was significantly greater (p < 0.001) than in the contralateral, non-swollen arm (-2.6 cmH2O range -11 cmH2O to 0 cmH2O). Interstitial fluid pressure in the oedematous arm did not correlate with the duration of the swelling (1-324 months), but did correlate with the increase in volume relative to the normal arm (r = 0.38, p < 0.05), the slope ('apparent compliance') being 28 ml.dl-1.cmH2O-1. The pressure-volume curve was less steep than the classic curve for acute oedema of dog limbs (Guyton, 1965). Vascular pressures were normal. The interstitial fluid pressures were not as high as those reported for lymphoedema of the lower extremity (mean 17.9 mmHg, Christensen et al., 1985). Nevertheless, the rise in interstitial fluid pressure by an average of 4.6 cmH2O constitutes a force opposing further microvascular fluid filtration and perhaps promoting fluid drainage out of the arm.


Tissue-type plasminogen activator and von Willebrand factor plasma levels as markers of endothelial involvement in patients with Raynaud's phenomenon.

In order to investigate the significance of high circulating levels of von Willebrand factor (vWF), recently observed in patients with vascular diseases, we compared the plasma levels of vWF with those of tissue-type plasminogen activator (t-PA) and the platelet content of serotonin (5-HT) in 40 patients with Raynaud's phenomenon (RP), primary or associated to systemic sclerosis (SSc), and in 14 patients with chronic peripheral obstructive disease due to arteriosclerosis (PAOD). VWF and t-PA plasma levels were significantly increased (p < 0.001) in SSc (vWF: 158.2, range 116.3-305.0%; t-PA: 10.2, range 6.4-17.8 ng/ml). By contrast, normal plasma levels of both vWF (85.3, range 53.5-157.0%) and t-PA (6.5, range 2.7-9.3 ng/ml) were observed in primary RP. VWF and t-PA were normal in PAOD patients, compared with age-matched healthy controls (vWF: 143.0, range 57.0-204.0%; t-PA: 7.5, range 3.4-13.6, ng/ml). The platelet content of 5-HT was within the normal range (37.3-99.7 ng/10(8) platelets) in RP patients, but significantly reduced (P < 0.05) in PAOD patients (39.0, range 14.7-91.4). Our data suggest that the different pattern of circulating vWF and t-PA between SSc and arteriosclerotic patients may be related to a different endothelial cell involvement. Whether this increase may reflect active attempts of regeneration and repair, indicating endothelial cell viability rather than damage is a matter of speculation.


Circulatory adaptation to the increased metabolism in the skin at the site of the tuberculin reaction.

The sequence of changes at the site of a positive tuberculin test response were studied in 19 healthy young adults who had been immunised with BCG in childhood. The development of erythema preceded that of induration and both were most intense at 48-72 h. The strongest reactions showed higher laser Doppler (LD) flux at the periphery than at the center (central relative slowing). All showed a substantial reduction in steady-state (ss) tcpO2 from 24 h onwards and the oxygen consumption rate (mlO2.kg-1.min-1), calculated from the rate of fall in tcpO2 during temporary cuff occlusion of arterial input, was raised (greater than two-fold) throughout the period of study (to 96 h). The density of lymphocytes and macrophages in the inflammatory infiltrate in the dermis was related to the fall in tcpO2.ss and to the extent of thickening of the dermis. These experiments showed that the previously healthy dermal microcirculation can adapt to temporary increase in metabolic demands of leucocytes emigrated from the circulation into the tissue: in intense delayed hypersensitivity (DHS) reactions there is considerable hypoxia and respiratory debt, but maintenance of viability in the short-term. It is likely that similar adaptations occur in the period of establishment of microbial infection.


Red cell velocity and microvessel diameter measurement by a two fluorescent tracer method under epifluorescence microscopy: application to cerebral microvessels of cats.

Fluorescein isothiocyanate (FITC)-labeled red blood cells (RBCs) and rhodamine-B isothiocyanate (RITC)-labeled dextran were used as two fluorescent tracers under intravital fluorescence microscopy. RBC velocity in cerebral microvessels of cats was measured with a dual window technique using FITC-labeled RBCs as a flow tracer. Measurement of the vessel diameter was performed using a digital image processor system by staining plasma with RITC-labeled dextran. The RBC velocity, obtained directly with digital cross-correlation of videodensitometric signals derived from the two windows, coincided with those obtained with the frame-by-frame analysis. The obtained RBC velocity ranged between 12.9 and 0.45 mm/sec in arterioles (< 60 microns) in diameter, 4.2 and 0.18 mm/sec in venules (< 60 microns in diameter), and 1.1 and 0.26 mm/sec in capillaries. The use of the labeled RBCs as the flow tracer enabled us to measure a wide range of RBC velocity (up to about 7 mm/sec) by setting the distance between the two windows beyond about 200 microns.


In vivo responses of allografted cerebral parenchymal arterioles to ethanol and angiotensin II: effect of calcium channel blockade.

The goals of these studies were to determine: 1) the effect of ethanol and angiotensin II on the diameter of allografted cerebral parenchymal arterioles in vivo, and 2) the effect of the calcium antagonist, verapamil, in modulating the responses of allografted cerebral parenchymal arterioles to ethanol and angiotensin II. Using a chamber technique, neonatal (< 24 hours old) cortical tissue was transplanted onto the cheek pouch of adult hamsters. Eight to thirteen days after allografting, hamsters were anesthetized with sodium pentobarbital (6.0 mg/100 grams i.p.), and allografted cerebral parenchymal arterioles were viewed using intravital microscopy. Diameter of allografted cerebral parenchymal arterioles was measured before (control), during and after topical application of ethanol (0.1%, 0.25%, 0.5%, 1.0%, and 2.0%) and angiotensin II (0.1 and 1.0 ng/ml). Application of ethanol and angiotensin II was repeated after changing the suffusion fluid to one containing verapamil (50 mg/L). We found that ethanol and angiotensin II produced dose-related constriction of allografted cerebral parenchymal arterioles. In addition, verapamil significantly attenuated vasoconstriction produced by ethanol and angiotensin II. Thus, our findings suggest that ethanol and angiotensin II cause constriction of allografted cerebral parenchymal arterioles through a calcium-dependent mechanism.


Xanthofibroma of the adrenal gland.

The authors report on a 43-year-old woman who had undergone unilateral adrenalectomy for a right suprarenal mass. The removed specimen was histopathologically defined as xanthofibroma, a hitherto unpublished adrenal tumour. The connective tissue protein of this rare tumour contained great amounts of collagen, non-collagenous protein and elastin. The significance of such abnormal proliferation of fibrous tissue and the conventional sonographic and CT features of this neoplasm are discussed.


ICN/3M scholars: tomorrow's nursing leaders.

For over 20 years ICN and the Minnesota Mining and Manufacturing Company (3M) have sponsored a scholarship programme directed toward helping nurses further their education in nursing. Since then 48 nurses have been provided the needed funds to reach their professional goals and subsequently to help answer their country's healthcare needs. Their degrees and expertise have opened up professional doors but most of all have given them the confidence to become driving forces in initiating research and other programmes that aim to provide quality care to all their countries' citizens.


Inverted papilloma of the renal pelvis.

The authors describe a case of inverted papilloma. It is relatively rare and generally appears as a benign tumour.


Nursing in Africa today.

In any description of nursing in Africa, one must consider the continent's vast size and complexity of cultures. Even the concept of nursing does not have the same meaning in all African countries. Below, Osei-Boateng updates the status of African nurses, derived from her paper that appeared in the 10th anniversary brochure of the West African College of Nursing (WACN). To simplify the analysis, she focuses only on the region south of the Sahara--excluding South Africa--in her overview of the slow progress and vast education needs of nurses caught in the web of their countries' socioeconomic and political chaos ... and lethargy.


Multiple fibroepithelial polyps of the renal pelvis and calyces: a case report.

Fibroepithelial polyp of the renal pelvis is an extremely rare entity. We report a case of multiple fibroepithelial polyps of the renal pelvis and calyces and discuss this rare and confusing condition with its clinical, radiological and pathological findings.


A unique concept of nursing care.

The form of caring may vary across cultures; but the perception of the caring by the receiver is universal. The patient knows what hurts, what problems are crucial and what experiences have been deeply buried. As a Chinese from Taiwan, raised in Japan and educated in the US, the author provides a unique concept of nursing care that relies on the patient for direction.


Usefulness of non-invasive diagnostic methods for early detection of neoplasms of the urinary tract and male genitals among workers of the Mazovian Refining and Petrochemical Plant in Płock.

The studies were aimed at the evaluation of the usefulness of non-invasive diagnostic methods including inquiry, clinical examination, transabdominal ultrasonography, cytologic examination of the urinary sediment, determination of blood serum concentrations of certain neoplastic markers, and urinalysis for early detection of neoplasms of the urinary tract and male genitals; and at the determination of the sensitivity and specificity of percutaneous ultrasonography combined with cytologic examination of the urinary sediment for detection of neoplasms of the kidneys and urinary tract, particularly of the urinary bladder. Urological examinations were performed in 300 workers occupationally exposed to oil derivatives. Neoplasms of the urinary tract and male genitals were detected in 37 (12.3%) workers. Neoplasms of the urinary bladder were found most frequently. They were diagnosed by non-invasive methods in 14 (4.7%) and verified clinically and histologically in 11 (3.7%) persons. The results seem to indicate the usefulness of non-invasive diagnostic methods for urological screening studies among occupational populations with increased risk of neoplasms of the urinary system and male genitals. The high sensitivity (93%) and high positive predictive coefficient (0.90) for the combination of transabdominal ultrasonography and cytological examination of the urinary sediment for detection of urinary system neoplasms seem to be tokens of their high diagnostic value and clinical usefulness.


Participatory research: getting the community involved in health development.

The Alma Ata Declaration states that "the people have the right and duty to participate individually and collectively in the planning and implementation of their health care." This means that community involvement in health development is fundamental in the battle for health. And, as the author demonstrates below, its participation in health research is necessary in the process of finding better ways and methods to provide health for all.


In situ prone ESWL for the treatment of lower ureteral stones: experience with 28 patients.

Twenty-eight patients with lower ureteral stones underwent in situ extracorporeal shock wave lithotripsy (ESWL) in the prone position over the period of 7 months between March 1990 and September 1990. For stone disintegration the spark gap shock wave lithotripter Tripter XI (Direx) was used. Satisfactory disintegration was achieved in 93 per cent of patients. The stone-free rate at 12 weeks was 82 per cent, and 11 per cent had residual fragments less than or equal to 4 mm in diameter. Twenty-one per cent of patients required repeat treatments. For only 2 patients general anaesthesia was required (7 per cent). There were no remarkable complications except for haemospermia which resolved spontaneously 15 days after treatment. It was concluded that in situ prone ESWL is an effective and safe procedure for the treatment of lower ureteral stones.


Fifty years of antireflux ureterovesicoplasty.

In spite of the impressive progress in medical science, surgery remains an empirical skill. Surgery of the ureterovesical junction is a permanent challenge for all generations of urologists. Half a century of intensive development of modern surgery evidences the efforts to reestablish function by a perfect copy of natural anatomy.


The permanent ureterocutaneostomy (UCST).

During a period of eleven years, ureterocutaneostomy (UCST) as a permanent urinary diversion was performed in 44 highly selected patients. Postoperative complications, in particular stenoses or strictures, can be avoided if subtle operative techniques are applied. Ureterocutaneostomy, which is functionally similar to an incontinent ileal conduit, turns out reasonable especially as a palliative procedure in patients with unresectable pelvic mass, dilated ureter, or solitary kidney. Thus, the authors conclude that UCST as a permanent urinary diversion still has to be considered valuable in a highly selected group of patients.


Bladder leiomyosarcoma: report of three cases.

Three patients with leiomyosarcoma of the urinary bladder are presented. One case showed urothelial carcinoma in situ also. The mode of presentation and operative findings were similar in two female patients, whereas the lesion seemed to be more aggressive in a male patient.


Percutaneous drainage of prostatic abscess.

The treatment results of 2 patients with prostatic abscess who underwent perineal percutaneous drainage under transrectal ultrasonographic guidance are described. Both patients were treated definitely and without complications. It is concluded that the use of transrectal ultrasound during the procedure increases the effectivity and safety of treatment.


Prostatectomy in the very aged.

The number of very aged men who demand prostatectomy for benign disease is increasing. We have assessed surgery results in 79 patients operated during the last four years. Sixty-two underwent endoscopic and 17 open procedure. Mortality rate was 3.7%. Two patients died in the first week and one patient three weeks after surgery. Morbidity rate was 61% but did not significantly affect the final operative outcome. Operative success rate six weeks postoperatively was 87%, satisfactory for this age group. After detailed preoperative evaluation and postoperative care by specialized age care team, prostatectomy is safe, effective and involves low-cost treatment.


Prognostic variables in patients with prostate cancer: influence of blood group ABO (H), the Rhesus system, age, differentiation, tumour stage and metastases.

Among 279 patients with carcinoma of the prostate no relationship was observed in survival between blood groups ABO (H) or the rhesus system. Age at diagnosis is relevant for the outcome of the disease, but grade of differentiation, tumour stage and metastases are more important predictors of survival.


Update of experience in the treatment of localized carcinoma of the prostate by definitive external irradiation.

In 1987 a retrospective analysis was performed of 116 patients with carcinoma of the prostate localized to the pelvis who were treated by radiotherapy at the Northern Israel Oncology Center during the period between 1977 and 1982. It was shown that definitive external beam irradiation is an effective, simple, and easy treatment modality for localized prostatic cancer.


Psychosomatic evaluation of patients with congenital penile deviation. A postoperative catamnestic follow-up.

Thirty-three patients with congenital penile deviation were examined by a semi-structured interview and psychodiagnostic tests (Giessen test, Giessen complaint sheet) prior to undergoing surgery according to the Nesbit-Kelâmi technique. A prognosis was then made to determine the extent to which the problems and disturbances reported by the patient or uncovered by the examiner could be improved by the operation. This prognostic assessment was then verified in a subsequent catamnestic examination (N = 20), at an average of 21.3 months after the intervention. An attempt is made to identify and define preoperative indicators that permit to predict the postoperative course.


Prostaglandin E2 in renal transplant recipients.

The concentrations of prostaglandin E2 (PGE2), sodium, potassium and creatinine were determined in the blood and urine of 50 renal transplant recipients treated with cyclosporine A or azathioprine, as immunosuppressive agents, for at least one year post transplantation. Fourteen healthy subjects served as control group. The urinary PGE2 excretion was significantly decreased in all treated renal transplant recipients and this reduction was associated with significant decrease in urinary excretion of sodium and potassium. On the other hand, a high elevation in blood PGE2 concentration was observed while no significant changes were seen in sodium and potassium levels in the blood of these renal transplant recipients. The observations suggest an association between urinary PGE2 reduction and immunosuppressive treatments in renal transplant recipients; also, PGE2 may regulate intra-renal haemodynamics and influence the renal tubular electrolyte excretion. Moreover, urinary PGE2 can be used as an indicator of successful renal transplantation.


Vitamin B6 requirements in chronic renal failure.

According to our results the long-term daily oral supplementation of 6 mg vitamin B6 was sufficient for prevention of vitamin B6 deficiency in chronic renal failure, regular dialysis treatment and CAPD groups of patients. Haemodialysis and charcoal haemoperfusion have led to non-significant decrease of erythrocyte vitamin B6. A favourable effect was found of daily oral administration of 50 mg pyridoxine on electrophoretic mobility of peripheral blood lymphocytes and cellular immunity.


Laparoscopic surgery in urology.

In recent months, laparoscopy has been used for such urological procedures as varicocelectomy, pelvic lymph node dissection and even nephrectomy. It would appear that laparoscopic surgery is going to make an important contribution to the field of endourology. In order to assess the current status and value of laparoscopic techniques to urologists, we have reviewed the history of laparoscopy in general and urologic laparoscopy in particular.


Spontaneous peripelvic extravasation of urine.

Spontaneous peripelvic extravasation of urine is relatively uncommon. We experienced 11 cases of spontaneous peripelvic extravasation. Urinary obstruction was caused by calculi in 9 cases, invasion by sigmoid carcinoma in 1 case and ureteral tumour (transitional cell carcinoma) in 1. Six patients presented with signs of peritonitis and 4 out of 9 with calculus did not develop microhaematuria. It is important to differentiate peripelvic extravasation from other acute abdominal conditions. We must pay special attention to patients with signs of peritonitis and normal urinalysis.


Urinary tuberculosis: experience of a teaching hospital in Syria.

This study was designed to show our experience of urinary tuberculosis in one of the large teaching hospitals in Syria. It was a prospective study involving 48 patients (29 males and 19 females) with confirmed tuberculous lesions in the urinary tract. The study period was between 1982-1987. The presenting symptoms were protean, and there were often delays between onset of symptoms and eventual diagnosis. The highest age incidence was in the second and fourth decades. Beside the suggestive clinical manifestations, final diagnosis was reached by various means. Repeated examinations of EMU smears were positive in about 20% of cases. Urine culture was positive in 33.3%, while varied percentage of cases showed some radiological changes suggestive of tuberculosis. Other investigations included cystoscopy and biopsy of suspected lesions. Treatment was conservative by chemotherapy in 45% of cases, with complete recovery in about 33%, while acceptable results were shown in congruent to 19%, and failure of treatment or recurrence of active disease in 48% of cases. Some forms of surgical intervention were carried out in 55% of patients. Cure was seen in 50% of them, while 25% had acceptable results and failure was shown in the remaining 25%. It is concluded that urinary tuberculosis remains an important infectious disease problem in our country. The high rate of failure of both medical or surgical treatment is mainly due to late diagnosis.


Experience with the Direx Tripter X-1 shock-wave lithotripter.

The authors give account of the experience they gained with the Direx Tripter X-1 (ESWL) apparatus. In a random material they succeeded to render 81.5% of the patients stone free within three months. They judge the apparatus suitable for in situ treatment of pelvic and ureteric stones and for the monotherapy of staghorn calculi. It is easy to handle, inexpensive to buy and maintain but noisy to operate, moreover the patient has to be anaesthetized. All in all it is highly useful in the treatment of renal lithiasis.


Late development of pelviureteric junction obstruction (PUJO) in a girl with previously normal pyelogram: a case report.

A case of ureteropelvic junction obstruction in a girl with previously normal pyelogram is presented. The aetiology of this development is unclear but this report emphasizes the need for a close follow-up of patients with persistent flank pain even if the initial pyelograms are normal.


Transurethral ultrasonography, fiberoptic cystoscopy and bladder washout cytology in the evaluation of bladder tumours.

The impact of transurethral ultrasonography, fiberoptic cystoscopy and bladder washout cytology on assessment of bladder tumours was investigated in this study. Transurethral ultrasonography had an accuracy rate of 96.5% in diagnosing and staging bladder tumours. The accuracy rates of fiberoptic cystoscopy and washout cytology were 90.9% and 73.7%, respectively, in diagnosis. The efficacy of transurethral ultrasonography was found to be high enough for routine employment in the evaluation of the bladder tumours. Fiberoptic cystoscopy in conjunction with washout cytology as a combination relatively easy to perform can be used especially for follow-up purposes of the bladder tumours.


Terodiline in the treatment of nocturnal enuresis in children.

The effect of terodiline was investigated in 17 children with nocturnal enuresis. Twelve of them were evaluated urodynamically before treatment and 6 of these were proved to have detrusor instability with or without provocation. Terodiline in a daily dose of 12.5 mg was administered for 4 weeks. After treatment the number of wet nights per week has significantly decreased. Urodynamically, maximum cystometric capacity has statistically increased and in 2 patients involuntary contraction has disappeared. During these periods no serious side effects were encountered. Terodiline could improve urodynamic parameters in nocturnal enuresis. These changes appear to be favourable in clinical improvement.


Primary lymphoma of the penis with rationale of treatment.

Appropriate care of a patient with primary lymphoma of the penis requires consideration of the diagnoses, thorough evaluation, and knowledge of the various therapeutic approaches. A patient is presented with brief review of similar cases together with the rationale for the use of radiation therapy and chemotherapy rather than surgery.


Positron emission tomography in the study of acute radiation effects on renal blood flow in dogs.

Renal blood flow measurements have been carried out by means of positron emission tomography (PET) to facilitate the detection of radiation-induced injuries. The advantages of the method employed in animal experiments are described.


Central catecholamine, sympathetic nerve and vascular protein in the acute phase of two-kidney, one-clip renovascular hypertension in rats.

Incorporation of 3H-proline into the non-collagenous protein in mesenteric arteries in two-kidney, one-clip hypertensive rats was greater than that in normotensive rats. Splanchnicotomy predominantly over the root of mesenteric arteries or intracranioventricular injection of 6-hydroxydopamine prevented the development of hypertension in 2K-1C rats concomitant with the reduction of incorporation of 3H-proline into the non-collagenous protein in mesenteric arteries. The content of norepinephrine in the hypothalamus in 2K-1C rats was lower than that in normotensive control rats. These findings indicate that increased non-collagenous protein synthesis in mesenteric arteries or low level of hypothalamic norepinephrine has facilitative effects on the development of 2K-1C hypertension.


The variety of clinical and histopathologic presentations of glomerulonephritis associated with latent syphilis.

Glomerulonephritis is a well established but rather uncommon complication of latent secondary syphilis. We present three cases of glomerulopathies associated with luetic infection, observed and managed in our institutions in the past three years. They illustrate a variety of clinicopathologic presentations of this nephropathy, from acute nephrotic syndrome through membranous glomerulopathy up to rapidly progressive glomerulonephritis. Regardless of the clinical course and histologic type, they were all characterized by strongly positive results of serologic tests for syphilis. Our observations suggest the necessity of eliminating luetic infection in aetiologic considerations of each newly diagnosed case of nephrotic syndrome.


Calcium blockers enhance cisplatin-induced nephrotoxicity in rats.

To examine the effects of calcium blockers on nephrotoxicity caused by cisplatin, the renal function and renal accumulation of Pt in Sprague-Dawley rats given 6.5 mg/kg i.v. cisplatin simultaneously with several doses of verapamil or nicardipine were evaluated. BUN, serum creatinine and kidney Pt concentrations in rats given more than 5.0 mg/kg of verapamil were significantly higher than those of the control animals injected with 6.5 mg/kg i.v. cisplatin alone, and the increase of each value was dependent upon the dose of verapamil. BUN, creatinine and kidney Pt in rats injected with more than 0.5 mg/kg i.p. nicardipine were also significantly higher than those of the controls. Calcium blockers enhanced the renal accumulation of Pt and the nephrotoxicity of cisplatin.


Influence of prostacyclin infusion on haemodialysis efficiency and blood cells.

Dialysis efficiency, platelet and leukocyte counts, as well as malonyldialdehyde (MDA) level and lactate dehydrogenase (LDH) activity in serum were assessed in 10 patients (8 males, 2 females, aged 28-58 years) treated with repeated haemodialysis due to terminal renal failure. Patients were examined twice: during a 4-hour haemodialysis in the presence of heparin as the anticoagulant, and a week later in the course of another haemodialysis combined with infusion of heparin and prostacyclin. Statistically significant lower level of urea at the end of dialysis and significantly higher urea clearance were found during haemodialysis with prostacyclin-heparin infusion in comparison with infusion of heparin alone. As compared with the initial values obtained prior to dialysis, neutropenia and thrombocytopenia were observed during haemodialysis with heparin alone but the counts remained generally unaltered when both prostacyclin and heparin were administered. During and after haemodialysis with heparin and prostacyclin both MDA level and LDH activity were lower than in case of haemodialysis with heparin alone.


Peritoneal dialysis in Hungary.

In 1991 the technical conditions and the number of patients receiving peritoneal dialysis were surveyed in the Hungarian nephrology and dialysing units. Not only the number of patients with chronic uraemia (undergoing dialysis + transplantation) is lower in this country as compared to the European average (106 and 236 per one million people, respectively), but also their distribution according to the type of treatment is different. For several years patients under intermittent peritoneal dialysis make up more than 10% of the cases and those under continuous ambulatory peritoneal dialysis less than 2% (in Europe: < 2% and 4-43%, respectively). The survey also included the types of solution, disinfection and connecting devices used in peritoneal dialysis, as well as the incidence of peritonitis and the administration of antibiotics. The principles of biocompatibility, the function of interleukin, as well as the effectiveness and the conditions of continuous ambulatory peritoneal dialysis are summarized.


CAPD versus haemodialysis: a comparison in the same patients.

Most studies comparing CAPD and haemodialysis (HD) were done in different populations, matched for sex and age. The present report compared 13 non-diabetic end-stage renal failure patients who were treated for at least six months with each type of therapy. Analysis of the data revealed a higher haemoglobin during CAPD but no differences in the blood transfusion requirements. Serum creatinine, BUN and potassium were lower during CAPD and serum calcium was higher during HD. Serum cholesterol levels were higher during CAPD and returned to pre-CAPD levels during the fourth month after being transferred to HD. Hospitalization rates were similar with the two treatments. Our study confirmed previous sex- and age-matched studies comparing CAPD and HD therapy.


Adjustment problems among Soviet immigrants at risk. Part I: Reaching out to members of the "1000 families" organization.

An outreach, preventive mental health project with Soviet immigrants implemented during and immediately following the Gulf War is described. One hundred and fifty immigrant families were visited by immigrant mental health professionals when the level and nature of their psychological distress was assessed. In the analyses of data, patterns of distress over time and factors which contributed to psychological distress were also examined. The main finding of the study concerned the subjects' well-being which was found to be marginally related to the Gulf War. Furthermore, no post-traumatic reactions could be discerned. Instead, immigration related factors, both emotional and objective, were correlated with the psychological distress of the subjects.


The psychological profile of Jewish late adolescents in the USSR: a pre-immigration study.

Psychological well-being and the level of psychological autonomy were studied in a group of 109 Jewish late adolescents in the USSR. The subjects were not found to be more distressed than their American peers, but their dependency on their parents was significantly higher. The findings are discussed in the light of Soviet social values and child rearing attitudes and their impact on the adjustment of Soviet immigrant adolescents in Israel.


Clinicians' assessments of suicide risk: can self-report measures replace the experts?

This study examined the possibility of using standardized self-report measures as "stand-ins" for clinicians' assessments of suicide risk. Subjects were 252 new applicants to a military mental health clinic who completed a battery of questionnaires and underwent clinical interviews. The self-report measures of psychiatric symptomatology (general and depression), personality (impulsivity, anger), and cognitions (hopelessness, attitudes toward life and death) were not highly correlated with clinicians' assessments of suicide risk and some showed a non-linear association. These findings suggest that the two methods of assessment are not interchangeable and that clinicians base their assessments of suicide risk, at least in part, on factors not assessed by the above questionnaires.


Program and process: designing the physical space of a day hospital.

Programmatic changes in the delivery of mental health services demand a reevaluation of the space devoted to those services. A growing body of mental health literature has been reported concerning the need to involve multidisciplinary teams in the design of mental health facilities. The development of Day Hospitalization Units offers a team the professional challenge of dealing with the physical environment. The following case study describes a given physical space and the process of change therein; based on this study and other relevant examples, we will discuss the relationship between physical space and the process of change.


Traffic accident injuries of children: the need for prospective studies of psychiatric sequelae.

This paper presents a literature review of epidemiological data relevant to traffic accidents of children, including their personality and behavioral predisposition, social factors, association with risk taking behavior, rehabilitation, and incidence of post traumatic stress syndrome. The need for prospective studies of psychiatric sequelae of such accidents is highlighted. Three case vignettes are presented.


Is there a place for paradoxical "maneuvers" in conventional psychotherapy?

This paper reviews the literature concerning the use and theoretical background of paradoxical maneuvers in conventional psychotherapy. By demonstrating a clinical example we highlight the usefulness of such an approach and the necessity for further research in finding valid indications and contraindications for its use.


Evolutionary patterns in chromosome numbers in neotropical Lepidoptera. I. Chromosomes of the Heliconiini (family Nymphalidae: subfamily Nymphalinae).

Chromosome counts in meiotic metaphase plates in the gonads of 67 of the probable 68 species of mimetic neotropical heliconian butterflies (Nymphalidae), representing 1524 individuals in 617 subspecies and geographically separate populations from southern Texas to northern Argentina, revealed a consistent haploid number of n = 21 in the genus Heliconius (except for the most advanced species with n = 33, 37, 56, and 60) and n = 31 in the more primitive genera (Eueides, Dryas, Dryadula, Agraulis, and Dione), with a transitional genus (Neruda) showing three species with n = 28-32, 21-22 + 5-10 "microchromosomes", and 20-22 + 1-5 "microchromosomes". The genus Laparus, with a single polymorphic species doris, probably an offshoot of early Heliconius, shows wide karyotypic variation (n = 20-30, 38) sometimes even within a single individual. The two most primitive genera also show much variation: Podotricha has two species with n = 9 and n = 26-29; and Philaethria shows many phenotypically similar species, two with n = 29 and a still uncertain number (at least 3) with n = 88 (most common), 67-72 (most widespread), 62 (very restricted geographically), 52, 21, and 12. Several interspecific hybrids (Heliconius cydno x H. melpomene) showed normal chromosome pairing, while deficient pairing was seen in intersubspecific hybrids in Eueides tales and Heliconius sara. The importance of these results in the evolutionary study of polytypic tropical species is discussed.


Putative fragile sites in the horse karyotype.

After fluorouracil/5-bromodeoxyuridine synchronization and subsequent FPG-staining, the karyotype of 15 phenotypically normal horses displayed several breaks and gaps. Twelve bands 1q24, 4p12, 8q23, 11p12, 16q21, 17q21, 23q31, 23q32, Xp21, Xq22, Xq25 and Xq27 showed relatively frequent fragility. After thymidine/cytidine synchronization and subsequent GWL-banding the same horses display karyotypes without any fragility. Hence it is suggested that the above listed bands harbour folate and/or 5-bromodeoxyuridine sensitive fragile sites.


Frequency of multiple insemination in a natural population of Drosophila montana.

The frequency of multiple insemination was studied in a boreal Drosophila montana population using mother-offspring data for a sex-linked allozyme locus. Mating with respect to the marker studied was random. In crowded laboratory cultures the heterozygous offspring had higher viability than homozygotes, but no deviations from Hardy-Weinberg proportions were found in natural populations. In multiply sired progenies the males did not contribute equally to the progeny, but the proportion sired by the second male was 0.76. The estimated frequency of multiple inseminated females was 1.19 +/- 0.31, indicating that practically all the females carried sperm of at least two males. The estimate is the largest ever reported in natural populations of Drosophila. However, the interspecific comparisons may not be relevant, because the frequency of multiple insemination does not necessarily reflect the real lifetime frequency of multiple matings. The effects of local ecology and life history characters, e.g., the uniform age structure and the temporal patterning of matings, on the high degree of detected multiple inseminations are discussed.


Effects of genetic and environmental factors on the a-b, b-c and c-d interdigital ridge counts.

A study of 100 MZ (55 female and 45 male) and 97 DZ (50 male and 47 female) same-sexed twin pairs was carried out to analyse the genetic component of the variance of the a-b, b-c and c-d interdigital ridge counts by means of the Christian method. Especially for the a-b interdigital ridge count, we found it important to analyse both sexes separately. Our results suggest that the a-b count in males seems to be more influenced by environmental factors than the other counts. For females, the three interdigital counts seem to have a strong genetic component influencing their phenotypic expression. Factor analysis with VARIMAX rotation showed each interdigital ridge count to be genetically independent.


The expression of parathyroid hormone-related protein (PTHrP) in normal parathyroid: histochemistry and in situ hybridization.

The expression and localization of parathyroid hormone-related protein (PTHrP), a major factor responsible for humoral hypercalcemia of malignancy (HHM), was investigated in 14 cases of surgically resected normal parathyroid glands. For light microscopic immunohistochemistry, formalin-fixed and paraffin-embedded specimens were stained with avidin-biotin-peroxidase complex (ABC) using the anti-PTHrP monoclonal antibody (MoAb), 4B3. Four percent paraformaldehyde (PFA)-fixed and OCT compound-embedded specimens were used for pre-embedded immunoelectron microscopy. For in situ hybridization, 4% PFA-fixed, frozen sections were studied using a bromodeoxyuridine (BrdU)-labeled PTHrP cDNA probe. Immunohistochemically, 12 of the 14 cases were positive for PTHrP, which was observed mainly in the oxyphil and transitional oxyphil cells. The chief and clear cells, on the other hand, were faintly positive. Electron microscopically, secretory granules positive for PTHrP were observed in cells containing abundant mitochondria. Consistent with the PTHrP immunoreactivity, transcripts of PTHrP were observed also in the oxyphilic cells by in situ hybridization. Thus the production and secretion of PTHrP was shown by the oxyphil cell lineage in the normal parathyroid glands.


Comparison of 35S- and digoxigenin-labeled RNA and oligonucleotide probes for in situ hybridization. Expression of mRNA of the seminal vesicle secretion protein II and androgen receptor genes in the rat prostate.

The sensitivity of radiolabeled and digoxigenin-labeled RNA probes and synthetic oligonucleotide probes for the detection of seminal vesicle secretion protein II (SVS II) and androgen receptor (AR) mRNA was compared by in situ hybridization in paraformaldehyde-fixed cryostat sections of the rat prostate. Both genes are expressed in different amounts in the various prostatic lobes and contiguous glands. SVS II or AR RNA probes were either labeled with digoxigenin-11-UTP or [35S]UTP by in vitro transcription. A synthetic SVS II oligonucleotide probe was 3' end-labeled (tailed) with either digoxigenin-11-dUTP or [35S]dATP. Hybridized 35S-labeled probes were detected by autoradiography and digoxigenin-labeled probes by immunohistochemistry using alkaline phosphatase conjugated anti-digoxigenin antibody or gold-labeled antibody followed by protein A-gold and silver enhancement. Digoxigenin-labeled probes provided the same degree of sensitivity as their 35S-labeled counterparts for the detection by in situ hybridization of weakly and strongly expressed mRNA. Using both labeling methods, the SVS II RNA probes were more sensitive than the oligonucleotide probes and background labelling of the 35S-labeled oligonucleotide probe was high. The digoxigenin method produced less background with all probe types, hybridization signals showed higher resolution and results were obtained faster than with radiolabeled probes. The immunogold silver enhancement system provided the fastest detection of digoxigenin-labeled probes with a sensitivity and resolution similar to that provided by alkaline phosphatase anti-digoxigenin immunohistochemistry. It is concluded that digoxigenin probe labeling and detection provides a sensitive, reliable, and efficient alternative to radiolabeled probes for in situ hybridization of mRNA.


Versatility of anti-horseradish peroxidase antibody-gold complexes for cytochemistry and in situ hybridization: preparation and application of soluble complexes with streptavidin-peroxidase conjugates and biotinylated antibodies.

In previous studies we have employed a gold-labelled, affinity-purified polyclonal antibody against horseradish peroxidase (anti-HRP--gold) in the avidin-biotin peroxidase complex (ABC) technique and indirect labelled avidin-biotin methods. The gold-labelled antibody was used as final revealing reagent to replace the 3,3'-diaminobenzidine (DAB) reaction by immunogold silver staining. The anti-HRP--gold reagent proved to be advantageous since blocking of endogenous peroxidase activity in the tissue sections was not further required and staining of superior contrast and resolution could be achieved in paraffin sections. In the present study we have optimized this technique by combining the last two incubation steps, i.e. HRP-conjugated streptavidin and anti-HRP--gold. Different ratios of the two reagents were tested empirically to establish the conditions for the formation of a soluble complex with optimal staining properties. Quantitative evaluation by densitometry of the staining intensity showed that the soluble streptavidin-HRP/anti-HRP--gold complex and the indirect labelled avidin-biotin method employing the gold-labelled anti-HRP antibody performed equally well. Thus, the availability of this complex simplifies the streptavidin-biotin immunogold technique for immunohistochemistry, lectin histochemistry and in situ hybridization and further demonstrates the versatility of anti-HRP--gold complexes.


Identification of brush cells in the alimentary and respiratory system by antibodies to villin and fimbrin.

Brush cells represent a population of epithelial cells with unknown function, which are scattered throughout the epithelial lining of both the respiratory system and the alimentary system. These cells are reliably distinguished from other epithelial cells only at the ultrastructural level by the presence of an apical tuft of stiff microvilli and extremely long microvillar rootlets that may project down to the perinuclear space. In the present study we show that brush cells can be identified in tissue sections even at the light microscopic level by immunostaining with antibodies against villin and fimbrin, two proteins that crosslink actin filaments to form bundles. In brush cells, villin and fimbrin are not only present in the actin filament core bundles of apical microvilli and their long rootlets but, in addition, both proteins are also associated with microvilli extending from the basolateral cell surface of the brush cells. Basolateral immunostaining specific for villin and fimbrin does not occur in any other epithelial cell type of the respiratory and alimentary tract. Thus immunostaining with antibodies against both proteins allows unequivocal identification of individual brush cells even in sectional planes that do not contain the brightly stained apical tuft of microvilli and their long rootlets.


Neuron-specific enolase-like immunoreactivity in the vertebrate retina: selective labelling of Müller cells in Anura.

Neuron-specific enolase (NSE) immunocytochemistry was carried out in retinae of goldfish, axolotl, clawed frog, cane toad, lizard, chick, guinea-pig, rabbit, rat, cat and human. With the exception of Anura, strong immunoreactivity was seen in the large ganglion, amacrine cells and horizontal cells of the retina in all of the other species. Photoreceptors were found to be labelled in the rat and human retina and only one cone type in rabbit. Photoreceptor pedicles and ellipsoids were stained in the goldfish and the somata and inner segments of some photoreceptors in axolotl. In the axolotl retina, besides neurons, Müller cells (MCs) were also immunolabelled. In the retina of the cane toad and the clawed frog MCs were the only stained elements. Similarly in other parts of the central nervous system of the cane toad, glial elements of the optic tectum and spinal cord were immunoreactive. In contrast, in the peripheral nervous system, neurons of the 1st sympathetic ganglion and the 2nd dorsal root ganglion were labelled. In double-labelling experiments, glial fibrillary acidic protein and NSE showed colocalisation both in the glial elements of the optic tectum and spinal cord and in MCs of the retina of the cane toad.


Patching and capping of LFA-1 molecules on human lymphocytes.

The distribution and dynamics of LFA-1 molecules over the surface of human lymphocytes were analysed using immunogold label-fracture and fracture-flip methods. Patching and capping were induced by incubation at 37 degrees C with antibodies directed against the alpha and beta chains respectively of the heterodimeric LFA-1 molecule, and were followed by immunofluorescence. Treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA) to link LFA-1 molecules to the cytoskeleton increased the percentage of capped cells, implying a faster and more efficient process of capping. At all times of clustering or upon phorbol ester treatment, the concentration of LFA-1 in patches and then in caps was not accompanied by a parallel concentration of membrane particles on the freeze-fractured plasma membranes. Our results support the role of the cytoskeleton in regulating the capping phenomenon and in controlling the structural organization of the plasma membranes.


Does ex vivo labelling of proliferating cells in colonic and vaginal mucosa reflect the S-phase fraction in vivo?

The ex vivo labelling of DNA-synthesizing epithelial cells in colonic and vaginal mucosa was compared with in vivo labelling. For this purpose, in vivo S-phase cells were labelled with [3H]thymidine (Tdr) and ex vivo labelling was continued by culturing tissue specimens in bromodeoxyuridine (BrdU). Various methods of tissue culture were employed in order to improve diffusion of medium (and BrdU) in the tissue. BrdU and 3H-TdR labelling were evaluated by immunohistochemistry and autoradiography respectively. Ex vivo labelling resulted in a patchy distribution of labelled cells, which did not correspond with the 3H-TdR labelling pattern obtained in vivo. Under the described conditions ex vivo labelling does not appear to be a reliable for estimation of the proliferative activities in vivo.


Vimentin antibodies stain membranous epithelial cells in the rabbit bronchus-associated lymphoid tissue (BALT).

The lymphoepithelium covering the bronchus-associated lymphoid tissue (BALT) of the rabbit lung was studied with monoclonal antibodies against vimentin, using the indirect immunoperoxidase technique. In the lymphoepithelium single cells which had a membranous apical cytoplasm and engulfed intraepithelial lymphocytes were vimentin-immunoreactive. All other epithelial cells of the lymphoepithelium and of the surrounding airway epithelium did not bind vimentin antibodies. The results support the hypothesis that the membranous epithelial cells in the lymphoepithelium of rabbit BALT are analogous with intestinal M-cells, which in rabbit Peyer's patches and appendix are selectively labelled by vimentin antibodies.


Transplantation in the treatment of paralysis agitans (Parkinson's disease).

Over the past 3 years, there has been great interest in transplantation therapy in the treatment of Parkinson's disease. Following the impressive results reported by Madrazo in the spring of 1987, more than 350 cases of adrenal medullary implantation have been performed worldwide. There has been a significant reduction in "off" time and an increase in "on" time without chorea in 40% of patients having this procedure. The duration of effect is 1 year in half of these cases, with the other half (20% of all patients) still demonstrating significant improvement 3 years after the procedure. The mechanism of the bilateral beneficial improvement is unknown. The survival of adrenal medullary tissue has not been demonstrated at autopsy. It is thought that the mechanism of improvement involves either regenerative sprouting of the remaining dopamine producing neurons as a consequence of the release of neurotrophic factors or an interruption of the striatal pallidal output inhibitory influence of the basal ganglia on the thalamus (or both). Fetal mesencephalic implantation has also been attempted in more than 100 cases worldwide. The improvements when seen are not any more dramatic than those following the best results of adrenal medullary implantation. Graft survival has not been proved; it remains a possibility that interruption of the putamino-subthalamic pallidal pathway or a trophic influence of the tissue provides an alleviation in parkinsonism. The ethical controversy, need for long-term immunosuppression, and difficulty with obtaining tissue of the appropriate age and delivering the appropriate quantity to the putamen have made this technique less than adequate.(ABSTRACT TRUNCATED AT 250 WORDS)


Managing a patient with progressive HIV disease.

This monthly series was developed from the AOA Task Force on AIDS Writers' Workshop, held August 16 to 18, 1991, in New York. The workshop was sponsored by an education grant from Burroughs Wellcome. It will provide brief clinical updates and perspectives on the human immunodeficiency virus (HIV). Readers may request tear sheets from the AOA editorial offices.