[Gerhard Henrik Armauer Hansen--still of current interest].

Armauer Hansen described his first observation of Mycobacterium leprae on 28 February 1873 in detail. His discovery of the leprosy bacillus was the result of a logical process in which distinct stages can be clearly seen. The first step was clinical, and established criteria for leprosy as "a specific disease". The second stage was epidemiological. Contrary to the main view favouring a genetic basis, these observations convinced him of the infectious nature of the disease. The third stage was the search for the bacillus. His basic observations were reproduced, as documented. Armauer Hansen afterwards tried to fulfill Koch's postulates, in order to prove that the bacillus was the causative agent of leprosy. His second main contribution is the work on leprosy control, emphasizing isolation to contain the infection. The subsequent decline of leprosy in Norway differs from the trend in many developing countries, where leprosy has remained an important public health problem. In recent years Norwegian medical research has made significant contributions to leprosy research, in the field of epidemiology in Bergen, and through immunological studies of the disease originating from the Armauer Hansen Research Institute in Addis Ababa, Ethiopia.


[Leprosy and heredity. Reflections of a district physician in 1884].

During the 19th century leprosy was a serious health problem in Norway, especially in some western, rural districts. In 1856 it was decided that all leprous patients should be examined by the local doctor (District Health Officer), and registered in a national leprosy register. The patients' family relationships received special attention. Some patients tried to avoid registration, fearing that the data might be misused. After Armauer Hansen (1841-1912) discovered in 1873 that leprosy was an infectious disease, isolation of leprous patients was enforced. In 1884 Thomas Collett (1835-1898), the local doctor in a rural district of western Norway, carried out a survey of all leprous patients registered in his district, a total of 164 patients. The data from his survey provide convincing support for the view that hereditary factors play an important role in the development of the disease. Modern research has confirmed that an important gene controls the susceptibility to leprosy.


[No connection between scarlet fever and gouty fever. Historical analysis from Ytre Nordhordland during 1862-1884].

In 1987 there was an unexplained increase in severe streptococcal diseases in Norway and other western countries. In Norway this increase was not accompanied by a corresponding increase in acute rheumatic fever. This study investigated the occurrence of scarlet fever and acute rheumatic fever in a rural district (approximately 15,000 inhabitants) of western Norway during the years 1862-1884. Four epidemics of severe scarlet fever occurred during this period. The local doctor treated 1,155 patients (96% children), of whom 154 (13.3%) died. Acute glomerulonephritis with subsequent kidney failure seems to have been a major cause of death. During the same period 76 patients (96% adults) were treated for acute rheumatic fever. These cases were not related to the severe epidemics of scarlet fever. It is probable that different, co-circulating strains of streptococci caused the infections, which were followed by glomerulonephritis and rheumatic fever. It is possible that rheumatic fever was caused by the strain that induced the more benign "Angina tonsillaris".


[La pierre de la folie. The stone of madness].

During the 16th and 17th centuries the expression "to have a stone in the head" was synonymous with peculiar behaviour or mental symptoms. Many quacksalvers earned their living by treating such patients with a scalp incision, pretending to remove a stone. Some of the great painters from this period have used the subjects as models for their works of art. A series of paintings of this kind have been traced in different museums, catalogues and publications, and are described, followed by a short survey of popular beliefs and superstitions connected with stones.


[New electronic methods for clinical measurement of temperature].

A proposed Norwegian ban on clinical thermometers containing mercury will affect the equipment available for clinical measurements of temperature. The basic technological principles and applications of electronic thermometers and infrared tympanic thermometers are reviewed and discussed.


[How are we today? On physicians' health, well-being and job satisfaction].

The authors present a review of Scandinavian and English literature on physicians' health and well-being. A simple model with three groups of effect variables (health, well-being and job satisfaction) and 16 groups of possible affect variables (gender, age, personality, family background, childhood conditions, student years, house officer period, working conditions, specialty, conditions and attitudes in society, patients, colleagues, lifestyle, living conditions, social network and family) are used as a framework for the review. Suicide mortality is higher among physicians than in other occupations. Female and young physicians seem to be most at risk of experiencing stress and psychosocial problems. There may be an inherent personality incongruence between the ideal doctor and the good spouse and parent. Having an "instrumental" specialty and working style, and not having to cope with too much professional uncertainty, seem to protect against stress and burnout. The review shows that greater consideration should be given to factors like these when carrying out a comprehensive study of physicians' health, well-being and job satisfaction. The forthcoming survey of Norwegian physicians' health, well-being and job satisfaction (Legekårsundersøkelsen) will include all the areas reviewed.


[Foreign physicians in Norway. Where do they come from--how do they work--what are their plans for the future?].

In a market with almost 500 vacant positions for doctors it is of considerable interest to obtain more knowledge about immigrant doctors. In a survey of 408 immigrant doctors, the majority came from the Nordic (22%) and other West European countries (37%), whereas the rest came from Eastern Europe (19%), Asia (17%), America and Africa. One third (36%) obtained their first appointment in central Norway and 22% in remote North Norway. The corresponding figure for the present appointment were 40 and 17%. Most doctors (73%) had been well received in Norway. Language problems (21%) and social isolation (16%) were frequently encountered, but professional problems were less common (7%). Doctors from West European countries complained that their medical qualifications tended to be under-rated by their Norwegian colleagues.


Contractile behaviour of skinned papillary muscle in mitral valve disease.

The contractile behaviour of Triton-X 100 skinned left ventricular papillary muscle from 19 patients undergoing cardiac surgery for mitral valve stenosis: n = 6, mitral valve incompetence: n = 7, or combined mitral valve disease: n = 6 was analyzed. At supramaximal activation the "vibration induced force clamping technique" was used for isometric analysis of time course and extent of isometric postvibration force recovery. Afterloaded contractions were applied for extrapolation of the maximum shortening velocity at zero load (Vmax). The Calcium sensitivity was analysed by variation of the free EGTA-buffered Calcium concentration at a passive resting force of 2 mN at 26 degrees C. In different types of mitral valve disease the characteristics of isometric force development were unaltered in terms of maximum force development, force per square mm, Calcium sensitivity and the time course of isometric contraction after force clamping. However the capability to shorten as expressed by Vmax was reduced in mitral valve incompetence (3.87 +/- 0.37 ML/s) as compared with mitral valve stenosis (5.29 +/- 0.35 ML/s) or combined mitral valve disease (4.83 +/- 0.51 ML/s). The ratio between the inverse value of Vmax and the time constant of isometric force development after force clamping was significantly different in mitral valve incompetence as compared with other types of mitral valve disease (p < 0.0001). These data argue for the presence of different resistances against shortening in various types of mitral valve disease, due to altered cross-bridge cycling characteristics or to morphological factors.


Total correction of tetralogy of Fallot in adolescents and adults.

From 1976 to 1988, 23 adolescent and adult patients underwent total correction of tetralogy of Fallot. There were 13 males and 10 females, ranging in age from 16 to 47 years (mean 24.3 +/- 8.6 years). Eight patients were in New York Heart Association (NYHA) functional class II, 14 patients in class III, and one patient in class IV. Sixteen patients (69.6%) had undergone previous palliative operation. All shunts were patent at the time of repair. In 9 patients bovine pericardial monocusp patches were used for reconstruction of the right-ventricular outflow tract. Intraoperatively, the right-ventricular to left-ventricular systolic pressure ratio after repair ranged from 0.29 to 0.80 (mean 0.49 +/- 0.13). There were 2 early deaths (8.7%). Eight of 23 patients (34.8%) exhibited postoperative low cardiac output syndrome. One late death occurred: a 22-year-old male patient died of Staphylococcus sepsis 8 months postoperatively. All surviving patients were followed from 3 to 15 years (mean 8.3 +/- 2.7 years). No patient required reoperation in the follow-up period. The actuarial survival estimate for all 23 patients was 87% at the end of 15 years. At follow-up 17 patients were in NYHA class I, two were in class II, and one was judged to be in class III. We believe advanced age is no contraindication to surgery in tetralogy of Fallot. Adolescents and adults remain in need of total correction which can be performed with acceptable risk and long-term symptomatic improvement.


Different ways of application of the implantable cardioverter defibrillator (ICD)--which is the current approach?

The implantable cardioverter defibrillator (ICD) has proved to be effective in patients at risk of sudden cardiac death (SCD). Because of the development of more effective lead- and defibrillator systems the surgical implantation modes have changed from extensive operative interventions (sternotomy, lateral thoracotomy) to less traumatical (subxiphoidal/subcostal approach) and less invasive (transvenous) techniques. Procedures with less operative damage may be offered to patients with reduced cardiac function. However, the kind of operative procedure to be performed must be decided individually. The different system configurations, their advantages and disadvantages as well as their complications are surveyed and discussed in the light of our patient population, 102 cases between 1984 and 1991, and the literature. Even when the "non thoracotomy" implantations will become the favoured method, the standard incisions by thoracotomy will still be of importance. Considering that major complications occur also in transvenous implant procedures and that cardiac operations have to be performed in 20-30% of the patients simultaneously, we suggest--following the recommendations of the North American Society of Pacing and Electrophysiology (NASPE)--that the devices be implanted exclusively in units for cardiac surgery in cooperation with rhythmologically well-trained cardiologists.


Haemodynamic and oxymetric investigations on patients under rate-responsive dual-chamber stimulation during exercise.

Rate-response dual-chamber pacing combines the advantages of atrio-ventricular synchrony and rate adaptation in patients suffering from binodal disease. However, it is not known if patients really benefit from the additional rate response under exercise conditions. Therefore, 15 patients suffering from binodal disease received rate-responsive dual-chamber pacemakers. 1, 3, and 6 months postoperatively haemodynamic parameters and oxygen consumption were measured during treadmill ergometry under conventional (DDD) and rate-modulated AV-universal stimulation (DDDR). Using the NCCOM3-R7 (Osypka) technique, rate (f), cardiac index (CI), stroke volume index (SVI), and the end-diastolic volume index (EDVI) were determined non-invasively. Oxygen consumption (VO2) was measured with the Biotec Oxysuper. Based on preoperative status (NYHA) and ventricular contractility, which was determined by means of echocardiography, the patients were divided into two groups: Group I (n = 8) with normal ventricular contractility demonstrated a moderate increase of intrinsic heart rate during exercise. Group II (n = 7) with impaired ventricular contractility showed only a slight or no increase in intrinsic heart rate. When comparing DDD and DDDR pacing a significant increase of CI and VO2 was found for the rate-modulated pacing mode in group II. The reverse was found for patients of group I. The difference between the two groups was statistically significant. According to these results, only patients with nearly no increase in intrinsic sinus heart rate and impaired ventricular function can be expected to benefit from the additional rate-adaptation of a dual-chamber pacemaker.


Cardiac surgery in HIV-positive intravenous drug addicts: influence of cardiopulmonary bypass on the progression to AIDS.

In the last years the number of HIV-positive patients needing cardiac surgery has greatly increased. Cardiopulmonary bypass is suspected to have a role in the progression of HIV-infection to acquired immunodeficiency syndrome (AIDS). From October 1988 to December 1990, 6 intravenous drug addicts underwent cardiac surgery for infective endocarditis at our Department. Preoperative and postoperative absolute lymphocyte T-helper (CD4) and T-suppressor (CD8) counts did not show a close association between the temporary lymphopenia induced by cardiopulmonary bypass and progression to AIDS.


Myocardial ischemia and left-ventricular function after ligation of left coronary artery (Bland-White-Garland syndrome): a long-term follow-up.

Left-ventricular function and myocardial perfusion were studied in 8 patients for between 3 and 21 years (mean 16 years) after ligation of the anomalous left coronary artery from the pulmonary artery. While only one patient complained of exercise-dependent stenocardia, 4 patients had abnormal ST-segments in the exercise ECG and all patients showed myocardial ischemia at exercise scintigraphy. At rest, the ejection fraction of the left ventricle was within normal range in all patients. In 5 patients, it failed to increase adequately under exercise conditions, reflecting impaired ventricular function. These results confirm the advantages of surgical procedures establishing a 2-coronary system.


Can the chest tube draining the pleural cavity with persistent air leakage be removed?

The pleural drain with persistent air leak in six selected patients after pulmonary surgery was clamped or removed without causing pulmonary collapse. In all the patients, air leak through the chest tube was present when speaking or coughing but not seen when breathing normally at rest or taking deep breaths. The fact that the chest tube could be removed in selected patients is explained by supposing the air leakage through an alveolopleural fistula is dependent on pressure difference between the alveoli and the pleural cavity, and this was confirmed in a rethoracotomy case for persistent air leak. Removal of the chest tube following the above-mentioned rationale would reduce the number of rethoracotomy cases for air leak and facilitate early removal of the chest tube in some patients.


Surgical approach to pseudoaneurysms with Behçet's disease.

One of the arterial complications of Behçet's disease, arterial false aneurysms, has a sudden onset of symptoms and high mortality risk if left untreated. Four cases with arterial pseudoaneurysms of 3 male patients is reported and the choice of surgical intervention is emphasized. It is underlined that PTFE grafts are the graft of choice in reconstructions for arterial complications of Behçet's disease.


Desmoid tumour in thoracotomy scar 5 years after excision of a left giant atrial appendage aneurysm in female with a family history of Gardner's syndrome.

A very large desmoid tumour extensively involving the chest wall and the left anterior abdominal wall is described in a patient with a family history of Gardner's syndrome. The desmoid arose at the site of a thoracotomy scar due to the removal of a large aneurysm of the left atrial appendage five years before. Both a plastic surgeon and a thoracic surgeon were required to remove the tumour. Aneurysmal dilatation of the left atrium is extremely rare and has not been reported in association with Gardner's syndrome before.


Is cellular disruption the mechanism of release of basic fibroblast growth factor from anterior pituitary gonadotropes?

Adult female Fischer 344 (F344) and Sprague-Dawley (SD) rats, intact and ovariectomized (10-30d), have been used for immunolocalization of basic fibroblast growth factor (FGF). Tissues were selected from three specific sites (postero-lateral, lateral wing, and anterior wedge) of the periphery of the anterior pituitary (AP) carefully maintaining the association between the gland and the highly vascular meningeal connective tissue which envelopes it. In both rat strains, most of the periphery of the AP was characterized by intact parenchymal cells delimited from the meningeal connective tissue by an intact basal lamina. However, foci also were evident in which parenchymal cells projected directly into the connective tissue without a basal lamina intervening. These zones, designated the Non-Delimited Peripheral Parenchyma (NDPP), were present minimally in control rats, but were more numerous in ovariectomized rats. Profiles of focally disrupted gonadotropes were evident within the NDPP of 20-30d ovariectomized rats juxtaposed against intact, granulated parenchymal cells. Partially disrupted gonadotropes also were evident within the peripheral parenchyma within approximately 100 mu of the edge, and occasionally the disruptions resulted in an association of neighboring gonadotropes as a syncytium. FGF was localized only within the cytosol of gonadotropes, i.e., cells immunopositive for LH-beta and FSH-beta subunits. Gonadotropes nearer to the edges of the AP, especially the postero-lateral edge, were the most intensely stained. Electron microscopy and immunostaining for S-100 protein, a marker for folliculo-stellate cells (FSC), demonstrated that in intact and ovariectomized SD rats FSC were present in all peripheral zones of the AP, whereas portions of the postero-lateral periphery of the AP of intact and ovariectomized F344 rats often lacked FSC. We propose that FGF may be released from the cytosol of gonadotropes by a mechanism of cellular disruption. FGF released at peripheral sites of the AP would be well-positioned to stimulate angiogenesis from systemic blood vessels within the meninges. Since FSC are known phagocytes within the AP, their consistent presence in the periphery of the AP of SD rats may help regulate the effects of the released FGF, and by contrast, their absence in F344 rats may intensify or prolong the effects of released FGF. Such differences may underlie the higher incidence of pituitary tumors in F344 rats.


Synaptic contacts established by primary sensory fibers innervating the teeth of rats: an ultrastructural study of the pars interpolaris of the spinal trigeminal nucleus.

Transganglionic degeneration was used in an electron microscopic study of afferent synaptic contacts in the dorsomedial region of the pars interpolaris of rats. In one experiment, the left inferior alveolar nerve was transected and in the other, partial pulpectomy of the first and second left lower molars was performed. Well defined degenerating terminals, almost completely occupied by round synaptic vesicle profiles were found in both ipsi and contralateral sides. In both experiments, approximately 70% of these terminals formed single asymmetric contacts with intermediate or distal dendritic segments. Fewer contacts were observed with proximal dendritic segments, dendritic spines, perikarya and other terminals. In addition, double and multiple synaptic contacts (synaptic glomeruli), accounting for 10% of the total, were also observed. Quantitative data regarding ultrastructural synaptic elements suggest that there is no preference for post-synaptic sites of fibers related to different sensory modalities such as pain, conveyed by dental fibers or other modalities, conveyed by the inferior alveolar nerve fibers.


Proliferative activity of cells of the intermediate lobe of the rat pituitary during the postnatal period.

Cellular proliferation was studied in the intermediate lobe (IL) of the pituitary gland of developing rats by labelling cells at the S-phase of the cell cycle with bromodeoxyuridine (BrdU). The number of BrdU-labelled cells in the IL decreased from birth until the 14th postnatal day and was low from that day until the end of the first month after birth. Throughout the postnatal period a large proportion of BrdU-labelled cells was found in the marginal layer (ML) of the IL, suggesting for the ML a role as a germinative layer of the IL during postnatal growth. Double immunostaining with anti-BrdU and anti-MSH showed that MSH cells actively proliferate as from the day of birth. Cells doubly immunostained with anti-BrdU and anti-S100 protein were first seen on the 14th postnatal day. From then onwards, most proliferating cells were labelled with either anti MSH or anti S-100 protein. This, together with the high proportion of proliferating cells found in the ML marks a clear difference with the pattern of cellular proliferation previously reported during a similar period in the anterior lobe of the rat pituitary.


Age-related changes in the morphology and function of the zona glomerulosa of the rat adrenal cortex.

The age-related changes in the morphology and function of rat adrenal zona glomerulosa (ZG) were investigated by coupled stereological and radioimmunological techniques. For this purpose 4-, 8-, 16- and 24-month-old rats were studied. Aging caused a notable lowering in the plasma aldosterone concentration and a marked decrease in both basal and ACTH- or angiotensin II (ANG-II)-stimulated secretion of collagenase-dispersed ZG cells. Plasma renin activity (PRA) underwent an age-dependent decrease, while the plasma level of ACTH displayed a significant rise. ZG and its parenchymal cells did not evidence any age-related morphologically demonstrable alteration in their growth, nor ZG cells showed any marked ultrastructural change, with the exception of a severe lipid-droplet repletion. This last finding is in keeping with the aging-induced decrease in the secretory activity of ZG cells, inasmuch as lipid droplets are the intra-cellular stores of cholesterol esters, the obligate precursors of steroid hormones in rat adrenals. ACTH and ANG-II are well known to be involved in the maintenance of the growth of rat ZG; thus, the combined impairment of ANG-II production (as evidenced by PRA lowering) and increase in ACTH secretion may maintain unchanged ZG growth during aging.


Differences in cell division and thymidine incorporation with rat and primate fibroblasts in collagen lattices.

Human and gorilla dermal fibroblasts, primate cells, suspended in a collagen lattice, do not divide for the first 3 days. In contrast, rat fibroblasts divide within 24 hr. In this study, the proliferation of rat fibroblasts were compared to primate fibroblasts. Rat fibroblasts in monolayer culture increase from 100,000 to 355,000 in 2 days, and human cells increase from 100,000 to 436,000 in the same period. An initial seeding of 100,000 rat fibroblasts suspended in collagen increased to 163,000 cells in 2 days. An initial 100,000 human fibroblasts seeded in collagen decreased to 80,000 cells in 2 days. Retarded proliferation of human and gorilla fibroblasts in collagen is unrelated to a defect in DNA synthesis. By autoradiography human fibroblasts suspended in collagen incorporate labelled thymidine. By flow cytometry analysis, the DNA concentrations of human fibroblasts suspended in collagen exhibited 41% in a 4N chromosome state, compared to 14% in monolayer culture. Nuclei of gorilla fibroblasts from collagen displayed 42% in a 4N state, compared to 19% in monolayer culture. With nuclei of rat fibroblasts from collagen, 14% were in a 4N state, compared to 9% in monolayer culture. Primate fibroblasts show a three-fold increase in the number of nuclei in a 4N state compared to rat fibroblasts suspended in collagen. After replating fibroblasts released from collagen in monolayer culture in the presence of 1 mM hydroxyurea (an inhibitor of DNA synthesis) primate fibroblasts doubled in 24 hr. Under identical conditions, rat fibroblasts showed no cell division.(ABSTRACT TRUNCATED AT 250 WORDS)


Airway branching patterns and cytodifferentiation in cultured fetal hamster lung.

Intact fetal hamster lungs were taken for culture on gestational day 12, when only lobar bronchi and primary bronchioles are established and the epithelial cells are undifferentiated. Explants were maintained on Transwell collagen membranes for 2 and 4 days in BGJb medium alone, with 5% FBS, or with the following additives: insulin, transferrin, hydrocortisone, cholera toxin, EGF, and vitamin A. Development of the respiratory tree was affected differently by each medium formulation. BGJb medium with 5% FBS permitted near normal branching of airways and presumptive alveoli. In contrast, BGJb medium alone permitted only limited branching of these structures. BGJb medium with additives permitted branching but markedly altered normal development. The differentiation of endocrine and secretory cells was monitored by immunolabeling for serotonin and calcitonin gene-related peptide, and Clara cell protein, respectively. Ciliated cells were identified by morphology. All medium formulations supported the timely differentiation of endocrine, secretory, and ciliated cells. The ultimate goal of our studies is to characterize factors that influence airway branching and cytodifferentiation during fetal lung development. This study showed that near normal airway branching and cytodifferentiation were supported in vitro by BGJb medium with 5% FBS. Although cytodifferentiation occurred with the two other formulations, airway development was impaired.


Female sex steroid induced epithelial changes in the gallbladder of the ovariectomized Syrian hamster.

Ovariectomized Syrian hamsters treated by female sex steroids during a 1-month period show gallbladder surface epithelial changes in the fundic area consistent with apical bulging and decapitations of the epithelial cells. These events were detected in the infundibulum and the fundic or body regions of estrogen- and estrogen+progesterone-treated hamsters. In control hamsters, these events were restricted to the region in the vicinity of the bile duct. Following steroid treatment, intraluminal deposits detected resembled Ca-bilirubinate deposits described in previous studies while decapitations are similar to endometrial epithelium changes associated with hormonal physiological changes or treatments. Moreover some small electron-dense deposits are comparable to those found in human cholesterol gallstones. This report indicates that, besides an alteration in bile composition, cell fragments originating from the surface epithelium of the bile duct and/or of the gallbladder mucosal epithelium could participate in gallstone nucleation.


Phenotypic responses of mouse mammary tumours and normal mammary epithelium cultured in collagen gels: correlation with tumour type and progression.

Mammary tumours in female BR6/Icrf mice and the corresponding contralateral normal mammary glands were disaggregated with collagenase and the epithelial structures released ('organoids') separated from other cellular components by filtration. The organoids were established in primary culture in a collagen matrix and the outgrowths obtained were studied by light microscopy and time-lapse cinemicroscopy. The pattern of three-dimensional outgrowths produced was found to be specific to the original tissue. Organoids from normal tissue formed a characteristic outgrowth designated Pattern A. Normal tissue from pregnant mice formed an additional characteristic outgrowth (Pattern A') which has not been described previously. Pregnancy-dependent tumours produced a distinctive phenotypic outgrowth designated Pattern D, whereas pregnancy-independent tumours gave a different distinctive Pattern B as well as a unique specific outgrowth designated Pattern C. Outgrowths of Pattern D from a pregnancy-dependent tumour were removed from culture and implanted into a syngeneic female mouse. Tumours arising in the host were found to be pregnancy-independent and showed phenotypic outgrowths in subsequent culture of pregnancy-independent Patterns B and C. The results show that the type of outgrowths in these cultures correlates with the biology of the tissue in vivo and that changes in tumour progression in vivo are accompanied by alterations in phenotypic outgrowths in culture.


A cytological and cytoskeletal comparison of Sertoli cells without germ cell and those with germ cells using the W/WV mutant mouse.

The distribution of F-actin and intermediate filaments in the W/WV mouse was investigated by light and transmission electron microscopy, and fluorescence methods. No spermatogenic cells were detected in the seminiferous epithelium of the W/WV mouse. Its seminiferous tubule was one-half the diameter of that in the normal (+/+) mouse. The Sertoli cell which was an only component of the W/WV mouse seminiferous epithelium was decreased in height, but still retained the polarity as evidenced by light microscopy. The Sertoli cell organelles were similar in appearance when normal and mutant mice were compared. F-actin was recognized at ectoplasmic specialization (ES) of the W/WV mouse Sertoli cell and appeared similar to the normal mouse. However, the junction with ES was more extensive compared with that of the normal mouse Vimentin in the W/WV mouse Sertoli cell was distributed around the nucleus and extended towards the tubular lumen similar to the normal mouse. Its extension within the Sertoli cell trunk, however, was restricted to a lesser degree as compared with that in the normal. Thus, the subcellular Sertoli cell and the distribution of F-actin and intermediate filaments (vimentin) in the W/WV mouse Sertoli cell seemed not to be strikingly affected by lack of spermatogenic cells, suggesting minimal influence of germ cells on Sertoli cell cytology and cytoskeleton.


Phagocytosis of collagen fibrils by periosteal fibroblasts in long bone explants. Effect of concanavalin A.

In an attempt to determine whether phagocytosis of collagen by fibroblasts involves binding of the fibril to the plasma membrane, the effect of the lectin concanavalin A (Con A) was studied in an in vitro model system. Metacarpal bone rudiments from 19-day-old mouse fetuses were incubated with varying concentrations of the lectin. Quantitative electron microscopic analysis indicated that Con A caused a dose-related increase in the amount of phagocytosed collagen fibrils in periosteal fibroblasts, suggesting either an enhanced uptake or a decreased intracellular breakdown of fibrils. Since a Con A-inducible increase was not seen in the combined presence of both the lectin and the proteinase inhibitor leupeptin, which is known to inhibit the intracellular digestion of phagocytosed fibrillar collagen, it is unlikely that Con A stimulated phagocytosis. Based on the finding that Con A interfered with the digestion of a synthetic substrate by the collagenolytic lysosomal enzyme cathepsin B it is suggested that the augmentation of intracellular fibrillar collagen under the influence of the lectin was due to a decreased intracellular digestion. Since Con A did not inhibit the uptake of collagen fibrils by the fibroblasts it is concluded that Con A-inhibitable binding sites for collagen molecules are unlikely to be involved in phagocytosis of collagen fibrils by fibroblasts.


Evaluation of four different immunogens for the production of snake antivenoms.

Four different immunogens were used to produce polyvalent antivenom in rabbits to the venoms of Bothrops atrox, Crotalus atrox, Crotalus adamanteus and Crotalus durissus terrificus. The immunogens were: (1) unfractionated mixture of the four crude venoms, and three fractions of the mixture as follows, (2) HPLC gel filtration high (> 50,000) mol. wt fraction, (3) HPLC gel filtration medium (14,000-50,000) mol. wt fraction, and (4) HPLC gel filtration low (< 14,000) mol. wt fraction. The resultant immune sera were compared with commercial antivenom (Wyeth, polyvalent Crotalidae) for total IgG content, ELISA reactivities, patterns of Western blots and ability to neutralize lethal and local hemorrhagic activities of the four venoms. The results indicate that the rabbit antivenoms had significantly higher ELISA reactivity and blotting signals than Wyeth antivenom. However, neither ELISA nor Western blotting signals correlated with the ability of the antivenoms to neutralize the lethal or hemorrhagic activities of the venoms. The protective ability of the antivenoms varied considerably. In general, antivenoms generated by using fractionated venoms as immunogens exhibited greater protective ability than antivenom produced by using the mixture of four venoms as immunogen. Some of the antivenoms provided greater or comparable protective ability for certain venoms when compared to Wyeth antivenom. It appears that the use of certain venom fractions as immunogens is a promising alternative for production of effective antivenoms.


Sequence determination and characterization of a phospholipase A2 isozyme from Trimeresurus gramineus (green habu snake) venom.

In addition to phospholipase A2-I (PLA2-I) reported previously (ODA et al., 1991, Toxicon 29, 157), a new PLA2 named PLA2-II was isolated from Trimeresurus gramineus (green habu snake) venom, and its amino acid sequence was determined by sequencing the native protein and the peptides produced by enzymatic (Achromobacter protease I and clostripain) cleavages of the carboxamidomethylated derivative of the protein. The protein consisted of 122 amino acid residues and His-47, Asp-48, and Asp-98 which have been assumed to be essential for PLA2 activity were conserved. Its sequence similarity to PLA2-I was 79%, with 26 residual differences. In contrast to the unique presence of Phe-28 in PLA2-I, PLA2-II contains Tyr-28 as seen in most of other PLA2s. There was no significant difference between the dissociation constants of PLA2-I and PLA2-II for Ca2+. Secondary structure compositions of PLA2-II were similar to those of PLA2-I and Crotalus atrox PLA2. A striking difference was found between these isozymes in contractile activity of isolated smooth muscle preparation of guinea-pig ileum. PLA2-II was over ten times more potent than PLA2-I, although its lipolytic activity toward egg-yolk was even slightly weaker (73%) than that of PLA2-I. The difference in contractile activities of PLA2-I and PLA2-II could be assumed to be due to discriminative lipid recognition brought about by different amino acid residues at the 58th position (Asp for PLA2-I and Asn for PLA2-II).


Isolation and physiological characterization of taicatoxin, a complex toxin with specific effects on calcium channels.

Taicatoxin is a new complex oligomeric toxin that was isolated from the venom of the Australian taipan snake Oxyuranus scutellatus scutellatus. It is composed of three different molecular entities: an alpha-neurotoxin-like peptide of mol. wt 8000, a neurotoxic phospholipase of mol. wt of 16,000 and a serine protease inhibitor of mol. wt 7000, linked by non-covalent bonds, at an approximate stoichiometry of 1:1:4. The most active form of the complex was isolated by ion exchange chromatography through DE-Cellulose followed by two steps of CM-Cellulose chromatography at pH 4.7 and pH 6.0, respectively. At this stage the complex migrates as a single component in beta-alanine-acetate-urea gel electrophoresis and is very toxic to mice (1 or 2 micrograms of the complex protein kills a mouse of 20 g within 2 hr). It blocks the high threshold calcium channel current of excitable membranes in heart and does not affect the low threshold calcium channel current. The block occurs at a site that is accessible extracellularly but not intracellularly. The block is selective for calcium channels, reversible, does not affect single channel conductance but only changes channel gating, and is voltage dependent with higher affinity for inactivated channels. The phospholipase activity of the complex toxin can be separated by affinity-chromatography using a phospholipid analog (PC-Sepharose). The resulting complex contains only alpha-neurotoxin and protease inhibitor and is still capable of blocking calcium channels, although with less potency than the native oligomeric form. Sephadex G-50 gel filtration chromatography in the presence of high salt (1M NaCl) at alkaline pH (8.2), separates the alpha-neurotoxin-like peptide from the protease inhibitor, but at this stage the resulting peptides lose physiological activity towards the calcium channels. The amino acid sequence of the protease inhibitor was determined by automatic Edman degradation. The alpha-neurotoxin-like peptide and two isosubunits displaying phospholipase activity were sequenced at the N-terminal part of the molecule.


Local heat and cold application after eastern cottonmouth moccasin (Agkistrodon piscivorus) envenomation in the rat: effect on tissue injury.

We studied the effect of local application of heat or cold on the development of tissue necrosis in envenomated rats. Anesthetized animals had 6 mg/kg venom from Agkistrodon piscivorus injected subcutaneously into the lateral aspect of a hind limb. Heat or cold was applied to the site of envenomation in the experimental groups for 4-6 hr, and the injected area was examined histologically after 24 hr. Neither local treatment, with or without the presence of systemic antivenin, significantly affected the severity of tissue necrosis induced by the venom in comparison to a control group left at ambient temperature.


A fibrinogen-clotting serine proteinase from Cerastes cerastes (horned viper) venom with arginine-esterase and amidase activities. Purification, characterization and kinetic parameter determination.

An enzyme displaying proteolytic activity toward the natural substrate casein as well as clotting activity on fibrinogen was purified to homogeneity from Cerastes cerastes (horned viper) venom and characterized. The enzyme is constituted of two identical subunits of mol. wt 48,500 as determined by SDS-polyacrylamide gel electrophoresis, and has an isoelectric point of 3.75. N-terminal sequencing up to the 33rd residue evidenced a high homology with other snake venom proteinases. The proteinase is of serine-type as indicated by high sensitivity to DFP and shows both arginine-ester hydrolase and amidase activities on synthetic substrates. Both specific activities were 30-fold higher than the respective activities found in the crude venom. The Km value determined for arginine-containing substrate BAEE was 3.0 x 10(-4) M and the Km for chromogenic substrate CBS 34-47 0.65 x 10(-4) M. The Vm/Km ratio, however, was two-fold higher for BAEE than for CBS 34-47; the arginine-esterase activity of this enzyme is thus slightly higher than its amidase activity.


Effects of canatoxin on the Ca(2+)-ATPase of sarcoplasmic reticulum membranes.

Canatoxin, a toxic protein isolated from the seeds of Canavalia ensiformis, was studied for its effects on the sarcoplasmic reticulum vesicles obtained from rabbit skeletal muscle. Canatoxin inhibited Ca2+ accumulation catalysed by the Ca(2+)-ATPase, without affecting the hydrolytic activity of this enzyme or membrane permeability to Ca2+. The effects of canatoxin were dose dependent, but not time dependent. It is concluded that canatoxin interacts with the Ca2+ pump and uncouples Ca2+ uptake from Ca(2+)-dependent ATP hydrolysis.


Role of Tyr and Trp in membrane responses of Pyrularia thionin determined by optical and NMR spectra following Tyr iodination and Trp modification.

Pyrularia thionin is a strongly basic and bioactive 47 amino acid peptide which contains two Tyr residues at positions 13 and 45 and one Trp at position 8. Limited iodination does not have a significant effect, but prolonged iodination of the peptide leads to progressive inactivation for all known cellular responses (Evans, J. et al. (1989) Proc. natn. Acad. Sci. U.S.A. 86, 5849-5853). 1H NMR spectra of the native Pyrularia thionin show four Tyr bands, two arising from each Tyr residue. One resonance band for the epsilon hydrogens of Tyr 45 disappears early during limited iodination and the band from the delta hydrogens shifts to low field. The two bands corresponding to Tyr 13 remain during limited iodination, but both decrease in intensity during prolonged iodination, with the epsilon hydrogen band decreasing somewhat more. The resonance bands arising from Trp disappear during prolonged iodination. This sequence of reactions is verified by the optical absorbance properties of two small peptide fragments obtained by Staphylococcal V8 protease hydrolysis of thionin which had been iodinated to varying degrees. Limited iodination did not significantly inhibit the thionin's biological activity, yet the fragment from the -COOH terminus showed the conversion of Tyr 45 to diiodoTyr. This treatment did not significantly modify the Tyr 13 or Trp 8 located in the -NH2 terminal fragment. More extensive iodination resulted in a disappearance of Trp 8 absorbance with an accompanying conversion of Tyr 13 to the monoiodo form. Extensive iodination yielded two atoms of iodine in the Tyr 45-containing fragment, and only one atom in the Tyr 13 fragment. The data indicate that Tyr 45 of the native thionin is more readily iodinated, proceeding to the diiodo form without significant loss of activity. Prolonged iodination does not lead to the formation of any diiodoTyr 13, but does lead to modification of Trp 8 and probably formation of monoiodoTyr 13. Modification of Trp 8 with N-bromosuccinimide inhibits the hemolytic activity of the thionin, showing that Trp 8 is necessary for Pyrularia thionin activity. It is most likely Trp 8 modification during prolonged iodination which results in the loss of biological activity.


An idiotypic-anti-idiotypic competitive immunoassay for quantitation of okadaic acid.

A competitive indirect enzyme-linked immunosorbent assay for the measurement of okadaic acid, a marine toxin, was developed. The assay uses a murine monoclonal anti-idiotypic antibody bearing an internal image of okadiac acid epitope to capture an anti-okadaic acid monoclonal antibody in the presence of free okadaic acid. Bound anti-okadaic acid antibody is detected with peroxidase-conjugated anti-mouse immunoglobulin antiserum. If present, free toxin will lessen the amount of anti-okadaic acid antibody binding to its corresponding anti-idiotypic antibody in a dose-dependent manner that can be quantified from the standard curve. The assay permits reliable measurement of okadaic acid in the 9-81 ng/ml range. The intra- and interassay coefficients of variation in the measurement of OA in the toxin spiked mussel samples averaged 9% and 12%, respectively. The assay is rapid, accurate, reproducible and relatively simple to perform. It may be of potential use to laboratories involved in monitoring the toxin levels in plankton, seafood or sponges.


Two new L-serine variants of microcystins-LR and -RR from Anabaena sp. strains 202 A1 and 202 A2.

Two new microcystins, [L-Ser7]microcystin-LR (1) and [L-Ser7]microcystin-RR (2), were isolated from a filamentous fresh water cyanobacterium (blue-green alga), Anabaena sp. strain 202 A1, along with the two major toxins, [Dha7]microcystin-LR (3) and [Dha7]microcystin-RR (4) and their minor components the D-Asp variants [D-Asp3,Dha7]microcystin-LR (5) and [D-Asp3,Dha7]microcystin-RR (6). Anabaena sp. strain 202 A1 also produced another new toxin, whose structure is tentatively proposed as [D-Asp3,L-Ser7]microcystin-XR (7), where X is a leucine homologue. Anabaena sp. strain 202 A2 produced one new microcystin, 1, and three known microcystins, 3, 4, and 5. The structures of the toxins were assigned based on their amino acid analyses, and fast atom bombardment mass spectrometry data.


Two methyl ester derivatives of microcystins, cyclic heptapeptide hepatotoxins, isolated from Anabaena flos-aquae strain CYA 83/1.

Cultured cells of Anabaena flos-aquae strain CYA 83/1, isolated from Lake Edlandsvatn, Norway, produced two microcystin mono-methyl ester derivatives (1 and 2) at the D-Glu unit in addition to microcystin-LR (3), [D-Asp3]microcystin-LR (4), microcystin-RR (5), and [D-Asp3]microcystin-RR (6). Structures of these compounds were assigned based on their amino acid analysis with a Waters Pico Tag HPLC system plus fast atom bombardment mass spectrometry (FABMS), including tandem FABMS, analysis on the two new microcystins, [D-Glu(OCH3)6]microcystin-LR (1) and [D-Asp3, D-Glu(OCH3)6]microcystin-LR (2). Toxicity data were not obtained for 1 and 2 because of the small amounts isolated from the cells.


Isolation and structures of microcystins from a cyanobacterial water bloom (Finland).

A hepatotoxic cyanobacterial (blue-green algal) water bloom was collected from a constructed water reservoir in Finland. The water bloom contained two cyanobacterial species, Microcystis aeruginosa and Aphanizomenon flos-aquae. Two hepatotoxins, 1 and 2, were isolated from extracts of lyophilized cells. The structures of 1 and 2 were assigned based upon their amino acid analyses on a Waters Pico Tag HPLC system and a chiral GC capillary column (Chirasil Val III), fast atom bombardment mass spectrometry (FABMS), high resolution FABMS, and tandem FABMS data. Toxin 1 was identical to a previously reported compound, [D-Asp3]microcystin-RR. Toxin 2 was new and was assigned the structure [D-Asp3]microcystin-YR.


Isolation and structures of five microcystins from a Russian Microcystis aeruginosa strain CALU 972.

Five microcystins were obtained from Microcystis aeruginosa strain CALU 972 isolated from a hepatotoxic water bloom collected in Lake Kroshnosero (Russia). The structure of a new toxin (1) was determined as [Dha7]microcystin-YR by amino acid analyses and fast atom bombardment mass spectrometry, and the toxins 2, 3, 4, and 5 were assigned the structures as [Dha7]microcystin-LR, [D-Asp3,Dha7]microcystin-LR, [Dha7]microcystin-RR, and [D-Asp3,Dha7]microcystin-RR, respectively, by direct comparison with authentic samples.


A zinc-protease specific domain in botulinum and tetanus neurotoxins.

Neurotoxins produced by Clostridium botulinum are classified into groups (A to G) based on their serological nature. They consist of two subunits, heavy and light chains, linked by one or more disulphide bridges. The light chain is responsible for the blocking of acetylcholine release. Amino acid sequences of light chains have already been reported for botulinum toxins types A, C, D and E. Five highly homologous regions are found between these four toxins. One of these homologous regions, sequence HELIHSL, shows strong similarity with the active site of zinc-proteases. We suggest that inhibition of acetylcholine release might be associated with this protease activity.


Experimental acute intoxication with ripe fruit of Karwinskia humboldtiana (Tullidora) in rat, guinea-pig, hamster and dog.

Previous acute toxicity studies with 'Tullidora' in mice showed mainly hepatic and pulmonary complications. We tested samples in rat, guinea-pig, hamster and dog, searching for the mechanism of death. A single oral preparation of ripe fruit of Karwinskia humboldtiana was given to all animals. Clinical signs and histopathology were reproduced in all species except dogs. We propose a selective toxicity of the toxins of this fruit to the lungs and liver.


Dendrotoxin-like effects of noxiustoxin.

Noxiustoxin from the Mexican scorpion (Centruroides noxius Hoffmann) is known to block neuronal K+ channels. Noxiustoxin facilitated acetylcholine release in chick biventer cervicis nerve-muscle preparations, but not in mouse phrenic nerve-hemidiaphragm preparations. Noxiustoxin displaced binding of a radiolabelled dendrotoxin from synaptosomal membranes from rat brain, with a Ki of 10(-10) M. It is concluded that noxiustoxin shares some pharmacological properties with the K+ channel blocking dendrotoxins.


An EDTA.Ca2+ complex inhibits the enzymatic activity but not the lethality of beta-bungarotoxin.

An EDTA.Ca2+ complex inhibits the phospholipase A2 activity of the presynaptic neurotoxin beta-bungarotoxin without affecting its lethal potency. The EDTA.Ca2+ complex induces a conformational change in the enzymatic active site region of beta-BuTx, as indicated by the suppression of the 340 nm tryptophan fluorescence peak. Modification of the enzymatic site without loss of toxicity supports the presence of separate loci for the two activities.


Age changes of Vipera berus venom amidolytic activity.

The study of age changes of amidolytic activity of Vipera berus venom is investigated using mixtures of chromogenic peptide substrates differing by detected groups. Quantities of venom (total protein content) and its proteolytic activity from snakes of different ages were compared. The venom composition of newly born adders was shown to be considerably different from the venom composition of young (12-month) adders of the same population. The next evolution of protease activity is weakly expressed.


Molecular karyotype analysis in Leishmania.

The advent of pulsed field electrophoresis has allowed a direct approach to the karyotype of Leishmania. The molecular karyotype thus obtained is a stable characteristic of a given strain, although minor modifications may occur during in vitro maintenance. Between 20 and 28 chromosomal bands can be resolved depending on the strain, ranging in size from approximately 250 to 2600 kb. The technique has revealed a striking degree of polymorphism in the size and number of the chromosomal bands between different strains, and this seems independent of the category (species, zymodeme, population) to which the strains belong. It appears that only certain strains originating from the same geographic area may share extensive similarities. This polymorphism can largely be accounted for by chromosome size variations, which can involve up to 25% of the chromosome length. As a result, homologous chromosomes can exist in versions of markedly different size within the same strain. When this occurs with several different chromosomes, the interpretation of PFE patterns appears difficult without prior identification of the size-variable chromosomes and of the chromosome homologies. DNA deletions and amplifications have been shown to account for some of these size modifications, but other mechanisms are probably involved; nevertheless, interchromosomal exchange does not seem to play a major role in these polymorphisms. These chromosomal rearrangements, yet in an early stage of characterization, exhibit two relevant features: they seem (1) to affect essentially the subtelomeric regions and (2) to occur in a recurrent nonrandom manner. Chromosomal rearrangements sharing the same characteristics have been identified in yeast and other protozoa such as Trypanosoma and Plasmodium. The significance of this hypervariability for the biology of the parasite remains unknown, but it can be expected that such mechanisms have been maintained for some purpose; genes specifically located near chromosome ends might benefit from rapid sequence change, alternating activation, or polymorphism of expression. The chromosomal plasticity could represent a general mode of mutation in these parasites, in parallel with genetic exchange which may be uncommon in nature. The molecular characterization of these rearrangements, the identification of each chromosome with the help of physical restriction maps and linkage maps, and the collation of such data on a number of strains and species should allow a significant progress in the understanding of the genetics of Leishmania, in particular as regards ploidy, generation of phenotypic diversity, and genome evolution. Finally, like other models, this is susceptible to improve our knowledge of DNA-DNA interactions and of the chromosome functional structure and dynamics.


Acute aspirin overdose: mechanisms of toxicity.

The case history described herein illustrates many of the salient clinical and laboratory features of acute salicylate overdose. Aspirin overdose remains a commonly used means of attempting suicide in young adults. This case demonstrates the role of the laboratory in the management of salicylate overdose. Medical treatment of several critical concurrent metabolic responses was dependent on frequent evaluations of serum electrolytes, blood and urine pH, blood PO2, blood PCO2, and blood HCO3- concentrations. In addition, essential to her excellent medical care was the ongoing close interaction between the clinical toxicology laboratory and the clinical staff.


Monoethylglycinexylide formation in assessing pediatric donor liver function.

Lidocaine metabolism to monoethylglycinexylide (MEGX) has been described as a novel method to assess liver function in adult transplant donors and recipients. While this assay appears to offer a number of advantages over existing liver function tests, limited work has been done to evaluate its potential in the pediatric population. This study evaluated MEGX formation in potential pediatric liver donors (n = 35) and a control group of children (n = 16). The mean MEGX formation was significantly higher in pediatric donors than in the control group (156 +/- 62 vs 106 +/- 33 ng/ml, p < 0.05). No correlation with age, total bilirubin, liver transaminases, or alkaline phosphatase could be made within each group. Significant differences in MEGX levels were noted when each group was compared to its adult counterpart. Both pediatric donors and controls had greater mean MEGX formation than has been reported for adult donors and controls (156 +/- 62 vs 127 +/- 61 ng/ml, p < 0.05 and 106 +/- 33 vs 72 +/- 36 ng/ml, p < 0.05, respectively). Drugs that alter lidocaine pharmacokinetics and their potential influence on MEGX formation were evaluated in the pediatric donor group. Donors exposed to hepatic enzyme-inducing drugs had a higher mean MEGX formation (187 +/- 60 vs 146 +/- 63 ng/ml). No significant differences were noted between donors receiving and not receiving vasopressors. In conclusion, the significant differences between pediatric and adult MEGX formation should be noted when establishing reference or normal ranges for this diagnostic test. Furthermore, concomitant drug therapy may significantly alter MEGX formation.


A within-subject analysis of carbamazepine disposition related to development in children with epilepsy.

The effect of aging on carbamazepine (CBZ) plasma level/dose ratio was evaluated retrospectively in 15 children who were receiving CBZ monotherapy and who were followed up for at least 3 years. Subjects of the study were selected from a population of roughly 4,500 patients attending our therapeutic drug monitoring service during a 12-year period. Results showed that the CBZ plasma level/dose ratio increases within subject during childhood, in agreement with data obtained in between-patient studies. However, the increase is not linear with age, the greatest modifications being observed between 9 and 13 years of age. Weight gain alone does not seem to explain this finding, implicating the involvement of complex physiological changes occurring during puberty.


Postmortem redistribution of morphine in rats.

Because morphine is often found in the postmortem (PM) blood samples of patients treated with or abusing the drug, its causation in death has to be considered. The possibility of PM redistribution of the opioid drug has not been studied previously. We treated adult Wistar rats with 4 mg/kg of morphine i.m. and measured cardiac cavity levels at the time of death, by killing (2 h postdose) 24 and 96 h PM. Morphine concentrations rose from 41.4 +/- 13.2 ng/ml at death to 111.9 +/- 66.6 ng/ml at 24 h (p = 0.00036), with no additional increase at 96 h (98.7 +/- 21.7 ng/ml). After death there was accumulation of endogenous substances cross-reacting with the morphine radioimmunoassay. Although these attribute only 3% to total accumulation of morphine, they may complicate interpretation of morphine "readings" in patients who had not received the drug. Our study suggests that after death there is substantial redistribution of morphine. Elevated PM levels cannot necessarily by interpreted as representing antemortem concentrations.


Variability in the disposition of ibuprofen enantiomers in osteoarthritis patients.

The pharmacokinetics of the enantiomers of ibuprofen have been investigated following oral administration of 300 or 600 mg of racemic ibuprofen four times daily to 45 patients with osteoarthritis. Fifteen of these patients also received single doses of 300 or 600 mg of racemic ibuprofen. Serum concentrations of R- and S-ibuprofen and urine concentrations of the stereoisomers of ibuprofen and its metabolites were measured by high-performance liquid chromatography. The fraction inverted (F(inv)) of the inactive R- to active S-ibuprofen was estimated by a urinary metabolite method. For the 15 patients in both the chronic and single dose studies, there were no significant differences in the clearance of R-ibuprofen or F(inv). The elimination half-lives of R- and S-ibuprofen were comparable for the single and chronic dosing studies. The area under the curve (AUC) values, 6-h trough concentrations, and average steady state concentrations of the R- and S-enantiomers were statistically different after chronic dosing. Despite considerable variability in the clearances in these patients, e.g., clearance (CL) of R-ibuprofen showed 28-49% CV, much less variability was seen in F(inv) (range 9-19% CV), implying that patients would receive similar effective doses of active S-ibuprofen in spite of large differences in kinetics.


Use of theophylline in neonates as an aid to ventilator weaning.

Respiratory diseases are major causes of morbidity and mortality in premature neonates. Theophylline has been utilized as an adjunct in facilitating ventilator weaning and in the management of apnea with or without bradycardia. Patient characteristics associated with improved outcome from theophylline have not been determined. The purpose of this study was to evaluate parameters associated with improved outcome in neonates with respiratory diseases receiving theophylline. The study population consisted of premature neonates that were studied retrospectively. Criteria for entry into the study were (1) less than 40 weeks gestation, (2) a diagnosis of respiratory distress syndrome (RDS), apnea of prematurity, hyaline membrane disease (HMD), or bronchopulmonary dysplasia (BPD), (3) dependence on intermittent mandatory ventilation, (4) failure to wean from the ventilator 24 h or more before the study, or (5) receiving theophylline. In this study, we found no correlations between time to wean from the ventilator and postnatal age at the time theophylline was initiated, 5-min APGAR score, and final theophylline serum concentration before complete weaning from the ventilator. However, there were significant negative correlations between birthweight and gestational age with respect to time to wean from the ventilator. The average theophylline serum concentration before weaning from the ventilator for this population of neonates was approximately 5-10 micrograms/ml, indicating that theophylline is not beneficial as an aid to ventilator weaning at serum concentrations < 10 micrograms/ml.


Pharmacokinetics of rapamycin: single-dose studies in the rabbit.

The pharmacokinetics of rapamycin was investigated in five New Zealand white rabbits following intravenous administration of 0.05 and 0.5 mg/kg rapamycin in a randomized crossover fashion. Whole blood concentrations of rapamycin were analyzed by high-performance liquid chromatography (HPLC). Model-dependent and -independent parameters were calculated. The volume of distribution at steady state and total body clearance increased significantly as the dose increased. Rapamycin pharmacokinetics appear to be nonlinear. The whole blood volume of distribution, especially at the higher dose, indicated distribution out of the blood component. The drug is not cleared rapidly, with a terminal half-life of > 13 hours as calculated by model-independent parameters. The 24-h whole blood trough concentrations of the drug are well within the analytical range of the HPLC procedure. This should permit trough level monitoring for therapeutic range studies involving the drug.


Comparative oxycodone pharmacokinetics in humans after intravenous, oral, and rectal administration.

The pharmacokinetics of oxycodone have been determined after single-dose administration by the intravenous (4.6-7.3 mg), oral (tablets, 9.1 mg and syrup, 9.1 mg), and rectal (30 mg) routes, in 48 patients undergoing minor surgery. There were no significant differences in the mean elimination half-lives between the intravenous (5.45 +/- 1.43 h), oral tablets (5.65 +/- 1.13 h), oral syrup (4.80 +/- 1.13 h), and rectal suppository (5.40 +/- 1.19 h) formulations of oxycodone. After intravenous administration, the mean plasma clearance of oxycodone was 25.5 +/- 10.1 L/h and the mean volume of distribution at steady state was 2.5 +/- 0.8 L/kg. The mean normalized area under the curve (AUC/D) obtained after intravenous dosing (48.2 +/- 30.2 micrograms.h/L/mg) was more than twice the AUC/D values obtained after the administration of oxycodone tablets (19.8 +/- 3.5 micrograms.h/L/mg), oxycodone syrup (17.5 +/- 5.3 micrograms.h/L/mg), and rectal suppository (20.3 +/- 5.1 micrograms.h/L/mg), indicating that the amount of oxycodone reaching the systemic circulation after the extravascular routes of administration was < 50% of that obtained after intravenous dosing. The mean absorption lag times after oxycodone tablets (0.52 +/- 0.33 h), oxycodone syrup (0.48 +/- 0.40 h), and rectal suppository (0.76 +/- 0.47 h) were consistent with the onset of pharmacological effects reported by the patients.


Pharmacokinetics of diltiazem in patients undergoing coronary artery bypass grafting.

The pharmacokinetics of oral diltiazem (DZ) were studied in 10 patients on the day before cardiac surgery and on the next day during surgery and cardiopulmonary bypass (CPB). Six patients were taking DZ 60 mg q.i.d. and four patients were taking DZ 90 mg q.i.d. All had been receiving DZ treatment for at least 3 months. The plasma concentration profile of DZ on the day before surgery was assumed to reflect the steady-state condition. DZ showed dose-dependent kinetics. On the day of surgery, the levels of total DZ (TDZ) at 15 min and 1 h after the initiation of CPB were significantly reduced (approximately 50%) when compared with the pre-CPB level. The levels of unbound DZ (FDZ) and the two major metabolites, N-demethyl DZ (MA) and deacetyl-DZ (M1) were not changed significantly by CPB. The plasma unbound fraction value increased sharply from 0.43 +/- 0.12 before the onset of CPB to a peak value of 0.83 +/- 0.12 during CPB, and returned to baseline level 24 h after dosing. We conclude that CPB decreases the TDZ concentration but has little effect on FDZ, MA, and M1 levels. The lack of effect of CPB on FDZ was related to the reduction of plasma protein binding of DZ.


Single and multiple oral dose fluvoxamine kinetics in young and elderly subjects.

The pharmacokinetics of fluvoxamine maleate was studied in two separate studies in healthy young and elderly subjects. In a single and multiple oral dose administration study, six healthy young subjects received an initial 50 mg oral dose followed by 50 mg tablets every 12 h for 28 days. In a second study, 13 elderly subjects received 50 mg tablets every 12 h for 28 days. Fluvoxamine peak plasma concentrations were reached in approximately 5-6 h following oral administration of single and multiple 50 mg doses to healthy young and elderly volunteers. The area under the curve (AUC) of fluvoxamine tended to be larger following multiple (873 ng.h/ml) as compared to single-dose administration (652 ng.h/ml). Also the terminal half-life after chronic dosing (22 +/- 6 h) tended to be longer than after single dosing. Steady-state plasma levels were obtained within 10 days' administration. The pharmacokinetics of fluvoxamine in elderly healthy subjects were no different from those recorded in young subjects. These results suggest that it is not necessary to adjust the dosage of fluvoxamine in elderly depressed patients, on the basis of pharmacokinetic arguments. Independent of the age group, approximately 3% of a dose was recovered as unchanged drug in the urine.


A radioimmunoassay for the new antiviral agent 1-beta-D-arabinofuranosyl-E-5-(2-bromovinyl)uracil.

A specific and sensitive radioimmunoassay (RIA) for the measurement of 1-beta-D-arabinofuranosyl-E-5-(2-bromovinyl)uracil (BV-araU) in biological fluids has been developed. The assay has a range of 2.5-1,000 ng/ml and 10-1,000 ng/ml for serum and urine, respectively, and has the sensitivity to detect 2.5 and 25 ng/ml of BV-araU in serum and urine, respectively. A satisfactory zero binding and sensitivity were obtained after an overnight incubation at 4 degrees C. Separation of the antibody-bound and free ligand was achieved by employing polyethylene glycol-goat anti-rabbit gamma globulin separant. A quantitative recovery of the exogenous analyte was obtained at all concentrations of BV-araU tested. The assay is specific for the parent drug and is not affected by the presence of its metabolite, BV-U (bromovinyl uracil) or serum components (nucleotides, nucleosides, or sugars). Intraassay coefficients of variation were 3.1-4.4% and 2.5-4.2% for serum and urine controls, respectively. Interassay variability was < 8.6% for all serum and urine controls. Linear regression analysis showed that the correlation between RIA and high-pressure liquid chromatography was excellent (r = 0.997). The ascending dosage studies have been analyzed by the BV-araU RIA, and results indicate that the values of area under the serum concentration-time curve increased proportionally with the administered dose of BV-araU up to 80 mg. Cumulative urinary excretion data showed that approximately 50% of unchanged BV-araU was excreted in the urine within 24 h.


A liquid chromatographic assay using a high-speed column for the determination of lamotrigine, a new antiepileptic drug, in human plasma.

A sensitive, specific and rapid liquid-chromatographic method for the determination of the new antiepileptic drug lamotrigine (LTG) in human plasma is described. The method involves the use of a commercially available 3-microns particle size normal-phase column and a microflow-cell-equipped ultraviolet detector. Extraction is carried out with ethyl acetate after alkalinization on a 100-microliters plasma sample containing LTG and 3,5-diamino-6-(2-methoxyphenyl)-1,2,4-triazine as internal standard. The residue is reconstituted with 50 microliters of ethanol, and 5 microliters of the final solution is injected into the column. Elution is carried out at 35 degrees C using n-hexane/absolute ethanol/35% ammonia (80/20/0.25 by volume) as mobile phase at a flow rate of 2.0 ml/min. Detection is at 313 nm. The chromatographic separation requires < 3 min and the sensitivity limit is < 0.1 mg/L. Recovery is 88-96.2%, whereas within-day and day-to-day coefficients of variation are between 4.1 and 7.7%.


Determination of free valproic acid: evaluation of the Centrifree system and comparison between high-performance liquid chromatography and enzyme immunoassay.

Valproic Acid (VPA) is an important drug for the treatment of several types of seizures because it has a wide spectrum of activity. Since VPA has an unusual nonlinear binding characteristic and a wide interindividual variation, monitoring of its free concentration can be helpful in patient management. The determination of unbound VPA is more difficult because an extra sample preparation step is needed and the concentration of free VPA is low. Free drug monitoring can assume a more important role if there is a refinement in the technology. A high-performance liquid chromatography (HPLC) method with isocratic elution has been established for the analysis of the 4-bromomethyl-7-methoxycoumarin (BrMMC) derivative of free VPA. This method has a better sensitivity, linearity, and precision than enzyme immunoassay (EIA). Ultrafiltration with the Centrifree system was evaluated for the sample preparation. The influence of centrifuge times, relative centrifugal forces, and the starting sample amounts on the final results of the ultrafiltration were investigated. There was a satisfactory correlation between the free VPA levels determined by the HPLC method and the concentrations obtained by EIA. The total and free VPA were determined on 100 samples from 36 patients. The total VPA levels were in a range of 25 to 208 micrograms/ml, free VPA concentrations ranged from 1.92 to 55.75 micrograms/ml with the free fractions from 7 to 37%.


Bayesian estimation of streptomycin pharmacokinetics.

Streptomycin dose requirements were determined in an 83-year-old man with renal impairment who was being treated for miliary tuberculosis. Concentration measurements were interpreted using a Bayesian parameter estimation program. Estimated creatinine clearance (1.1 L/h) was used as a starting value for streptomycin clearance, and volume was initially assumed to be 0.3 L/kg. Bayesian estimates of clearance were close to the starting value and declined from 1.4 L/h to 0.9 L/h during the course of therapy. Volume was higher than the initial estimate (0.4-0.5 L/kg), possibly due to the patient having a low albumin and being underweight. Satisfactory concentrations were maintained for several weeks with doses of 500 mg every 36-48 h.


Analysis of ofloxacin in hair as a measure of hair growth and as a time marker for hair analysis.

The distribution of ofloxacin (OFLX) along a single hair shaft was analyzed in detail for use as an index of hair growth and as a time marker for drug analysis in hair. A single hair obtained from each of seven subjects, who had taken OFLX for 1-4 days (total of 200-1,200 mg) 2.7-5.3 months before hair sampling, was cut into 1-cm-long portions successively from its scalp end. OFLX in each hair portion was measured by high-performance liquid chromatography with a fluorescence detector, and the distance from the scalp end of the hair portion containing OFLX was determined. Then the other 2-cm long segment of hair, which had the above-determined distance at its middle, was cut successively into 2-mm-long pieces and OFLX was determined in each piece. This procedure was repeated in a total of three to four hair strands collected from one subject. OFLX was observed to distribute only in one to three consecutive 2-mm-long pieces of hair, showing no large diffusion of OFLX along the hair shaft with time. Therefore, OFLX distribution may serve as a time marker for analyzing other drugs in hair. Hair growth rate could be thus estimated and ranged from 0.99 to 1.27 cm/month (1.12 +/- 0.11 cm/month, mean +/- SD) among individuals. The intraindividual variability of hair growth rate was 4.8-18.1% (10.3 +/- 5.1%) as coefficient of variation.


A review of 43 cases of tetanus neonatorum.

Forty-three patients with neonatal tetanus admitted to the Infectious Diseases Ward of the Istanbul University Faculty of Medicine Children's Hospital at Capa from 1982-1989 are presented. Thirty-two (74.4%) of the subjects were boys. The overall mortality was found to be 60 percent and the mean incubation period 6.1 +/- 20 days. The mean period between onset of symptoms and hospital admission was 1.7 days. Thirty-nine (90.7%) of the 43 patients were unhygienic home deliveries and only two cases were delivered with a midwife in attendance. None of the mothers of the children in our series had been immunized with tetanus toxoid. The incidence of neonatal tetanus in Turkey is on the decline, but a fraction of the population, the so-called urban poor, is still at high risk for preventable diseases. Every contact an unvaccinated person has with a health care professional should be viewed as an opportunity for tetanus immunization.


The evaluation of vaccination against measles at nine months of age (report of an epidemic).

Sixteen measles cases were studied during an epidemic that broke out in Etimesgut district of Ankara. Eight of these children had never been vaccinated against measles while the remainder had been vaccinated at nine months of age. In the sera obtained during the course of the illness, anti-measles antibody was not detectable in six vaccinated children and in four unvaccinated children. Upon observing the siblings of the subjects, it was determined that one out of three who had not been vaccinated against measles and three out of seven who had been vaccinated at nine months of age contracted the disease within a month. However none of the siblings who had been vaccinated against measles at 15 months contracted the disease. In our cases, although vaccination at nine months of age could not prevent measles, it resulted in a milder form of the disease. It seems that measles vaccine administered to infants at around nine months of age does not prevent the occurrence of the disease in many children.


The relationship between blood glucose and serum electrolyte levels in children with acute diarrhea.

The relationship between simultaneous serum electrolyte and blood glucose levels was studied in 119 moderately or severely dehydrated children with acute infectious gastroenteritis. Their ages ranged between two months to fifteen years. A positive correlation was found between blood glucose and serum sodium levels in the children with serum bicarbonate values below 15 mmol/L. This correlation is more prominent in cases of severe metabolic acidosis with serum bicarbonate values less than 10 mmol/L. No correlation was found between blood glucose, serum potassium and chloride levels. Our results show that a positive correlation between blood glucose and serum sodium levels is present only in cases with metabolic acidosis, which is contrary to classical knowledge.


The correlation between the clinical, laboratory and histopathological features of childhood membranoproliferative glomerulonephritis and response to treatment.

In this study, the clinical, laboratory and histopathological features of 50 children with membranoproliferative glomerulonephritis are reviewed. Age distribution varied from 5 to 15 years. The clinical presentation in the patients was nephrotic syndrome (24%), acute nephritic syndrome (20%) and nephritic/nephrotic syndrome (56%). Hypertension, macroscopic hematuria and hypocomplementemia were present in 40 percent, 58 percent and 34 percent of the patients, respectively. Light microscopic findings were as follows: glomerular lobulation (36%), mesangial sclerosis (20%), tubulointerstitial findings (36%), and crescents (26%). C3 (93%) was the most common immunofluorescence and IgM (86%), the most frequently encountered immunoglobulin. Response to treatment could not be anticipated by the initial clinical and laboratory features. Patients who did not have tubulointerstitial changes tended to have a greater response to therapy.


Two-dimensional and Doppler echocardiographic findings of congenital coronary artery fistula.

Three patients with coronary artery fistula confirmed by coronary angiography and surgery were studied using two-dimensional and Doppler echocardiography. Two of these patients had left coronary artery-right ventricle fistula. The proximal dilatation of the coronary arteries was visualized in all three patients by two-dimensional echocardiography. The course of the dilated right coronary artery and left circumflex artery was demonstrated by two-dimensional echocardiography in two patients. We were able to detect the disturbed flow at the drainage site in one patient by using the conventional Doppler echocardiography. In conclusion, it should be emphasized that two-dimensional and Doppler echocardiography can be useful methods in diagnosing coronary artery fistula.


Neonatal peritonitis.

A retrospective study was undertaken over an eight-year period to assess the mortality rate of 66 newborns who had undergone surgery in our clinic because of peritonitis. The mortality rate for the patients admitted in poor condition was 95.2 percent, whereas it was 45.5 percent for those in good condition. There was a 100 percent mortality associated with newborns that had hypothermia and with those that had severe respiratory difficulties, whereas it was 92.3 percent for low-birth-weight infants and 86.5 percent for dehydrated infants. All babies with sclerema neonatorum died. The newborns with white blood cell counts under 5,000/mm3 also did not survive (83.3%). Etiologically, congenital megacolon, meconium ileus and spontaneous gastrointestinal perforations were the most frequent anomalies leading to death (100%). In the newborns with gastrointestinal perforations, most deaths occurred in patients with perforations of the cecum, duodenum and stomach (100%). Mortality seemed to be greater in patients with complications (77.3%), and it rose to 83.3 percent for patients who had to undergo a repeat operation due to complications. The overall mortality rate was found to be 71.2 percent.


Bronchogenic cyst in a 14-month old boy (a case report).

A patient with a bronchogenic cyst in the mediastinum is presented. This rare cyst was found by chance in a 14-month-old boy suffering from respiratory distress due to partial compression of a bronchus. Ultrasonographic and tomographic examinations revealed a cystic mass measuring 3 x 3 cm in diameter located within the apex of the right lobe of the lung. Bronchogenic cyst should be considered when these signs present in early childhood.


Atrial natriuretic peptide and aldosterone in sick premature neonates.

Plasma atrial natriuretic peptide (ANP) and aldosterone concentrations were investigated in 25 sick premature neonates. Group A consisted of 11 premature neonates with sepsis and group B of 14 neonates with hyperbilirubinemia. In both groups, ANP and aldosterone levels were found to be higher than in the controls. Group A concentrations of ANP and aldosterone were also higher than in group B.


Duplication of the rectum resembling a juvenile polyp.

A five-year-old boy with a rectal mass mimicking a rectal polyp, which proved to be a cystic duplication of the rectum, is presented. In a child with painless rectal bleeding, a mass palpated during rectal examination is usually diagnosed initially as being a rectal polyp. However, the case presented revealed the possibility of rectal duplication.


Congenital coronary arteriovenous fistula in a ten-year-old boy with angina pectoris.

Congenital arteriovenous fistula (CAVF) is a rare cardiac lesion. Angina pectoris is uncommon in younger patients with CAVF. Fistula-related symptoms, complications of this anomaly and surgical complications have a strong correlation with the age of the patient. A ten-year-old male patient with angina pectoris in whom the diagnosis of CAVF was established, and who, following surgical ligation recovered, is presented.


Meckel Gruber syndrome: a case diagnosed in utero.

A case of Meckel Gruber syndrome is presented, diagnosed prenatally from the medical history of the mother which revealed a previous malformed stillborn with anencephaly, meningomyelocele, polydactyly and ambiguous genitalia. This was the first prenatally diagnosed case ever reported in Turkey. The clinical, computed tomography and postmortem findings and the related literature are reviewed.


Relationship between latency of brainstem auditory-evoked potentials and head size in dogs.

Cranium and brainstem dimensions were measured in 32 postmortem dog heads. Positive correlations were found between cranium length (CL) and brainstem length (BL) (r = 0.87), between cranium width (CW) and brainstem width (BW) (r = 0.83), and between cranium distance (CD = CL+CW/2) and brainstem distance (BD = BL+BW/2) (r = 0.91). Positive correlation coefficients were also found between CL and CW (r = 0.90), and between BL and BW (r = 0.85). It was concluded that head size accurately reflected brainstem size. A least squares estimation of the brainstem distance (BD) from CL and CW values was BD = 10.9 + 0.16 (CL+CW/2) (BD, CL and CW in mm). Brainstem auditory evoked potentials (BAEPs) and cranium dimensions were measured in 43 dogs (86 ears) with different head size, body size, sex and age. Wave form, absolute and interpeak latencies and correlation coefficients, relating latencies to cranium dimensions and body weight, were analysed. CL, CW, and CD were positively correlated with body weight (r = 0.93, 0.70 and 0.93, respectively), and CL, CW, and CD were correlated with age (r = 0.33, 0.52, and 0.40, respectively). BAEPs consisted of five distinct positive peaks (I to V). Secondary positive peaks following peaks I and II were seen in 60% (I') and 90% (II') of the recordings. Late waves were recorded in 90% (VI), 50% (VII), and 25% (VIII) of the recordings. Latencies increased with decreasing stimulus intensity level (from 90 dB to 10 dB hearing level, HL), especially for peaks I, II, V, and the I-V interpeak interval.(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of methods used for blood collection on plasma concentrations of luteinising hormone, testosterone, and cortisol in male dogs.

The effects of two putative stressors relative to the collection of blood, namely the environment of the treatment room and the pain associated with venepuncture, on plasma levels of luteinising hormone (LH), testosterone and cortisol were examined in six trained male experimental dogs. Blood samples were collected from the dogs in a treatment room as well as in the kennels (control), and by venepuncture as well as via an indwelling intravenous catheter (control). No significant influence of either stressor on plasma levels of LH, testosterone or cortisol was found. Plasma concentrations of these hormones varied considerably both between and within dogs. Mean (+/- SEM; n = 6) plasma concentrations were 4.3 +/- 1.0 micrograms/l for LH, 4.6 +/- 1.9 nmol/l for testosterone and 68 +/- 10 nmol/l for cortisol. It was concluded that the putative stressors, the environment of the treatment room and the pain associated with venepuncture, did not significantly influence plasma levels of LH, testosterone or cortisol in trained male experimental dogs. This conclusion implies that under the experimental conditions described, the validity of results will not be affected by the method of blood collection used.


Ethical issues in the ownership of companion animals: an empirical and normative analysis.

This article deals with the ethical aspects of companion animal ownership. Research was done by means of interviews (n = 40) and questionnaires (n = 871). Respondents were randomly chosen from Dutch telephone directories. Euthanasia, docking tails, ear clipping and sterilisation are discussed on the basis of the opinions of respondents. Owners of companion animals very often seem to have insufficient knowledge about animal behaviour. Companion animal owners as a group differ rather strongly in their moral opinions about the acceptability of certain actions towards animals. At the individual level also people are not always consequent in their reasoning as was illustrated by their opinions about ear clipping and the stopping of tail docking and vice versa. Respondents live in a community with animals and usually use a utilitarian balancing model. The different outcomes of the uses of this balancing model indicate how far the respondents are willing to adapt their life style to the needs of an animal; in other words to what extent intrinsic values and integrity are taken into account within the process of (re)creating a living community.


The relationship between homozygosity and the occurrence of specific diseases in Bouvier Belge des Flandres dogs in The Netherlands.

Since purebred dog populations represent closed gene pools, a relatively high level of consanguinity between individuals, and hence of inbreeding, is common. This case control study was conducted to establish the possible connection between the actual level of homozygosity due to inbreeding and specific diseases (flea allergy, osteochondrosis, laryngeal paralysis, neoplasm, autoimmune disease, hypoplastic trachea, and food allergy) occurring in the Bouvier Belge des Flandres breed. One hundred and sixty-eight animals referred to the Utrecht University Clinic formed the patient group. Each of the seven diseases was chosen because of the demonstrability of the diagnosis. Each animal was chosen because one of the selected diseases was established in that individual, and because its pedigree was complete. All dogs of the patient group were born between 1 January 1980 and 31 December 1985. A control group (n = 123) was randomly chosen from the total population of registered Bouviers born in the Netherlands in the same period. pedigree data were obtained from the Dutch Kennel Club. The extent of inbreeding was determined for all animals using Wright's inbreeding coefficient. The distribution of inbreeding coefficients in each patient group was compared with the distribution in controls. Inbreeding coefficients in the control group ranged from 0.0 to 0.406. Animals in which osteochondrosis, food allergy, autoimmune disease, neoplasm, or hypoplastic trachea was diagnosed had higher inbreeding coefficients than controls. It was concluded that in the Bouvier Belge des Flandres dog population examined, the level of homozygosity was positively correlated with occurrence of these diseases.


An epidemiological study of Fasciola hepatica in The Netherlands.

Transmission of F. hepatica under natural conditions was analysed in a three year programme. The variables used were the indirect haemagglutination (IHA) technique, worm establishment in tracer lambs and the population dynamics, infection rate and shedding pattern of Lymnaea truncatula. It is concluded that fluke eggs, infected snails and metacercariae on herbage can survive the winter in the Netherlands. Metacercarial availability was positively correlated to the amount of rainfall in the grazing period. The role developed eggs that survive the winter is important, because this results in earlier infections in the herd. The use of the serological diagnosis method IHA is important to detect F. hepatica infection in an early stage. Use of cellophane paper on floats is a useful method for determining the shedding pattern of cercariae from L. truncatula. It is concluded that collection of metacercariae on cellophane floats, inventarization of L. truncatula and its infection level are useful tools for the prediction of liverfluke infections.


Subcutaneous tissue reaction to polyethylene terephtalate-covered electronic identification transponders in pigs.

Polyethylene terephtalate-covered identification transponders were injected in 4 week old piglets to examine clinically and histologically the reaction in the surrounding tissue after 4, 7 and 21 days and 6 months. Inflammatory signs at the injection site were clinically noticed from 2 days onwards and gradually decreased after day 3. A second series of inflammatory events occured in some animals around day 7. Swelling was observed thereafter in a few animals. In the pigs slaughtered at day 4, all samples showed a layer of exudate and debris surrounding the transponder. Afterwards a fibrous capsule developed. The mean thickness of reactive tissue decreased significantly (p < 0.01) between days 4 and 7 and days 14 and 21, and remained unchanged between day 21 and 6 months. The mean (+/- s.e.) capsule thickness was 0.32 +/- 0.12 mm after 6 months. It is concluded that PET-covered transponders are encapsulated in fibrous connective tissue within 3 weeks after injection at the base of the ear. After 6 months the capsule around the transponder revealed no or only minor signs of inflammation.


An immunohistochemical study of bovine antral follicles, with special attention to non-atretic follicles with and without atypical granulosa cells.

Inhibin and oxytocin were immunohistochemically demonstrated in all non-atretic and light-atretic follicles > 2 mm from untreated and pregnant mare's serum gonadotrophin (PMSG)-treated heifers and cows. Immunostaining for luteinizing hormone (LH) and oestradiol was observed in all non-atretic follicles > 4 mm, but only in follicles from PMSG-treated cows. Inhibin and oestradiol immunoreactivity was restricted to the granulosa. Oxytocin and LH immunoreactivity was visualized in both the theca interna and the granulosa. Within the granulosa, LH immunoreactivity was mainly present in cells that were located near the basement membrane. Normal granulosa cells differed from atypical granulosa cells (AGCs) with respect to their ability to bind LH and oestradiol. It is concluded that immunostaining for alpha-inhibin, oxytocin, oestradiol and LH cannot be used as a marker of follicle quality to discriminate between non-atretic follicles with AGCs and non-atretic follicles without AGCs in mid-luteal bovine ovaries.


Comparison of an indirect haemagglutination assay and an ELISA for diagnosing Fasciola hepatica in experimentally and naturally infected sheep.

An enzyme-linked immunosorbent assay (ELISA) with somatic (S) or excretory-secretory antigens (ES) was compared with an indirect haemagglutination assay (IHA) for ability to detect antibodies against Fasciola hepatica in sheep. The specificity of both assays was determined by testing sera collected from sheep experimentally or naturally mono-infected with Fasciola hepatica, Haemonchus contortus, Ostertagia circumcincta, Cooperia curticei, Taenia ovis, Eimeria spp., Trichostrongylus vitrinus, Trichostrongylus colubriformis or Nematodirus battus respectively. With S or ES antigens the specificity of the ELISA was 98% or 95% respectively, whereas the specificity of the IHA was 86%. Antibodies directed against Fasciola hepatica were detected by the ELISA with S or ES antigens from 2 weeks after infection until the end of the experiment, whereas the IHA detected antibodies from week 3. We conclude that the ELISA with S antigens compares favourably with the IHA and can be used for the serodiagnosis of ovine fasciolosis in the Netherlands.


Total CK and CK-BB activity in serum from sheep with scrapie.

Total CK and iso-enzyme CK-BB activity was measured in serum from four sheep with scrapie and in serum from four healthy control sheep. Blood samples were taken weekly for about six months. There was a clear overlap between the total CK and CK-BB activity in serum from sheep with scrapie and that in serum from control sheep. Thus measurement of these enzymes does not aid the clinical diagnosis of scrapie.


Hepatic lipidosis in pregnant cows on a dairy farm.

A syndrome very similar to hepatic lipidosis is described in dairy cows during the dry period. After being sent to pasture the animals did not eat well for undetermined reasons. The disease phenomena were mainly observed in animals carrying twins. At post mortem examination severe falty infiltration was found in the 3 animals made available for post mortem examination. Increase of the energy supply to the dry cows by addition of maize silage to the ration prevented new cases.


The survival of Sarcocystis gigantea sporocysts following exposure to various chemical and physical agents.

Using in vitro excystation as a measure of viability, it was found that at 4 degrees C Sarcocystis gigantea sporocysts survived considerably better in tap water (85% excystation after 174 days) than in either 2.5% potassium dichromate (15% excystation after 174 days) or 2% sulphuric acid (0% excystation after 5 days). Although they were able to resist 48 h suspension at room temperature in most laboratory reagents and disinfectants tested, six (sulphuric acid, ammonia, methanol, ethanol, potassium hydroxide, sodium hydroxide, Medol) had substantial sporocysticidal properties. Further investigation with three of these showed that sporocyst excystation was reduced from 65% to less than 10% following contact with 2.5% sulphuric acid for 1 h or with 2% ammonia or 4% Medol for 4 h. Sporocysts were either killed or had their ability to excyst severely impaired by heating to 60 degrees C and 55 degrees C for 5 and 60 min, respectively, by exposure to ultraviolet radiation at a dose of 4000 ET, or by prolonged storage in water at 24 degrees C. Sporocysts exposed to either constant or intermittent freezing at -18 degrees C suffered a comparatively slow decline in excystation rate with time, as did those subjected to desiccation. The duration of survival of desiccated sporocysts was inversely related to relative humidity and after 245 days at 33% relative humidity and temperatures of 15 degrees C or 24 degrees C, 60% of such sporocysts excysted.


Infectivity of Moniezia benedeni and Moniezia expansa to oribatid mites from Ohio and Georgia.

Six species of oribatid mites belonging to four families and five genera were successfully infected by M. benedeni. They were Scheloribates laevigatus (Scheloribatidae), Exoribatula sp. cf. biundatus (Scheloribatidae), Xylobates capucinus (Haplozetidae) and Zygoribatula undulata (Oribatulidae) from Ohio, and Galumna ithacensis (Galumnidae) and Scheloribates lanceoliger (Scheloribatidae) from Georgia. Cysticercoids were also recovered from S. laevigatus infected with M. expansa. Among the six species of mites, S. laevigatus is the major intermediate host of M. expansa and M. benedeni worldwide, while the other five species have not been previously reported as transmitters of Moniezia spp. This is also the first report of any species from the genera Exoribatula and Xylobates as intermediate hosts of tapeworms. Different species of oribatid mites varied in their efficiency as intermediate hosts of M. benedeni. Moniezia benedeni had higher infection rate and infection intensity in S. laevigatus than in other species. Development in S. laevigatus was also more rapid than in the other species. Mite size was only one of the determinants of intermediate host efficiency. Storage at 4 degrees C for 1 to 5 weeks reduced the viability of M. benedeni eggs. Stored eggs had lower infection rates and infection intensity and slower development in S. laevigatus. They also had lower infection rates in X. capucinus and Exoribatula sp. cf. biundatus.


Haemonchus contortus resistance to ivermectin and netobimin in Brazilian sheep.

Suffolk, Texel, Hampshire Down and Ile de France sheep from the municipalities of Porto Amazonas, Piraquara and Araucaria in the State of Paraná, and Bagé in the State of Rio Grande do Sul were brought to Sobral, State of Ceará, to be used in a cross-breeding project. On arrival they had clinical signs of nematode parasitosis, and one Suffolk female died. The animals were treated orally with ivermectin (0.2 mg kg-1) and fifteen days later with netobimin (20.0 mg kg-1). Neither drug reduced the egg counts (measured in eggs per gram, EPG) significantly, and this suggested that the nematodes in the sheep were resistant to the anthelmintics used. Haemonchus contortus was the species involved. The egg counts were reduced after oral treatment with trichlorfon (100.0 mg kg-1). Haemonchus contortus larvae obtained from these animals before trichlorfon treatment and passaged through two nematode-free sheep were used in a further experiment. Twenty 6- to 9-month-old nematode-free lambs were infected with the H. contortus larvae (10,000 per animal) and after the infection was confirmed, were randomly divided into four groups of five animals. Group I was orally treated with ivermectin at 0.2 mg kg-1, Group II with oral netobimin at 20.0 mg kg-1, Group III with oral trichlorfon at 100.0 mg kg-1 and Group IV was a non-treated control. Egg counts and faecal cultures were taken before dosing on the day of treatment and seven days later when all animals were necropsied and the nematodes were collected from the abomasa and counted.(ABSTRACT TRUNCATED AT 250 WORDS)


Epidemiology of trichostrongylidosis in cattle: development of infective larvae in the second part of the grazing period and their contribution to infections in the same year.

A grazing experiment was performed in the Swiss midland region with 22 first-season calves which were grazed together from May to mid July when they were allocated to two equal groups, one receiving a morantel sustained-release trilaminate bolus (Paratect Flex) bolus, Pfizer, Zurich, Switzerland) and the other one remaining as an untreated control. Each group was moved to a clean pasture on 21 July. The larval contamination on the pasture with the control animals reached 4652 third stage larvae (L3) per kilogram of dry matter (L3 kg-1 DM) after 10 weeks. Parasitic gastroenteritis was observed in two calves of the control group in August, and mean serum pepsinogen levels exceeded 4000 mU tyrosine in this group in September. Eggs from Ostertagia sp. and Cooperia sp. that were excreted by the animals at the beginning of July and the end of August showed similar hatching rates of 94% and 99%, respectively. In the calves of the bolus group the egg output was reduced by 95% within 14 days of administration of the bolus. Larval contamination on the pasture of the bolus group was zero until the beginning of September when a slight increase occurred, reaching 793 L3 kg-1 DM in October. At the end of the trial the bolus-treated calves had a mean weight gain advantage of 18 kg (P < 0.05) compared with the controls. The results confirm that a second generation of larvae developed on pasture in August and September and caused parasitic gastroenteritis in first-year grazing calves in late summer. The relevance of the results for the prophylaxis of parasitic gastroenteritis in calves in mixed grazing systems with calves and older cattle is discussed.