[The surgical treatment of malignant stomach ulcers]. The authors analyse the specific features of the clinical picture, diagnosis, pathomorphological changes and surgical treatment in 52 patients with gastric ulcer which underwent malignant degeneration. It is noted that malignancy occurs most frequently in close vicinity to the ulcerative defect and rarely in the mucosa beyond the ulcer. The morphostructural reorganizations are determined to a certain degree by the peculiarities of extension of the tumor along the mucosa and the organ as a whole. The authors discuss the principles of the choice of the method for treatment according to the findings of pathomorphological examination of biopsy material and the results of emergency intraoperative examination. Even if the merest suspicion of malignancy of the ulcer arises, the operation should be conducted with observance of all oncological principles. Postoperative mortality was 3.8%, complications occurred in 7.7% of cases. Five-year survival was 84.6% in the absence of metastases and the presence of carcinoma cells within the limits of the mucosa and 12.5% among cases in which metastases were found during the operation. Attention is drawn to the principles of regular medical surveillance over patients receiving nonoperative treatment for gastric ulcer.


[The groups at risk for a recurrence of rectal cancer after radical operations]. The article discusses a group of patients with rectal carcinoma who underwent radical operations at the Scientific Research Institute of Proctology in 1982-1983. The terms and frequency of rectal carcinoma recurrences are analysed and the factors influencing these indices are studied. The results made it possible to form a group with a risk of a recurrence among patients with carcinoma of the rectum and suggest new schedules of regular medical observation over these patients with consideration for the risk group.


[The surgical treatment of benign smooth-muscle tumors of the rectum]. From analysis of the surgical treatment of 36 patients with rectal leiomyoma the authors suggest and substantiate the indications for various types of operative interventions according to the size and location of the tumor. The late-term results of the treatment were studied in all patients who underwent surgery and a brief review of literature on the methods of diagnosis and surgical management of benign smooth-muscle tumors of the rectum is given.


[Kidney cancer morbidity in a population group under continuous observation in dispensary care]. Information on kidney carcinoma case rate among a contingent kept under continuous observation and subjected to regular medical examination is discussed. The authors had 390 cases under observation which allowed them to estimate the case rate according to the patients' sex and age group. A 5.8-fold increase of the case rate in a period of 15 years was found. In males it increased from 0.31:1000 to 1.8:1000, and was highest in the 50-59-year age group. Among females the kidney carcinoma case rate increased 2.5-fold in 15 years; it was highest among females aged over 70 (3.20:1000) and was 2.6 times higher than the case rate in the 50-59-year age group and 4.5 times higher than that in the 40-49-year age group (0.32:1000 and 1.44:1000, respectively). Kidney carcinoma takes a steady 6th place in the morbidity structure. Ultrasonic diagnosis of kidney carcinoma is the most effective method. The authors point out that study of the case rate of malignant kidney tumors is very important for elaborating the volumes of examination.


[The characteristics of kidney cancer invasion of the major retroperitoneal veins]. The authors discuss the variants of kidney carcinoma thrombus invasion of the major retroperitoneal veins, up to the right atrium, on basis of the literature data and their own findings during prosectorial (4) and clinical (7) practice. They emphasize that carcinoma may not only invade the large veins in the cardial direction in the blood flow but may also spread against the flow into the contralateral renal, hepatic, iliac, and other veins, which is illustrated by examples. Symptoms of cardiovascular insufficiency with venous congestion in the liver and lungs with the formation of red infarcts, the development of ascites and even with fibrous changes in the organs and insufficiency of their function came to the forefront in mild clinical signs of the main focus of affection. According to the clinical material, the incidence of thrombi in the inferior vena cava was 3.9%. All the thrombi in operated on patients were below the diaphragm and measured up to 11 cm in length. Histologically the thrombus consisted of formed elements of the blood, tumor cells (adenocarcinoma of light cells) which were inside it and fibrin which was outside. The thrombi grow slowly and undergo organization and vascularization.


[Tendoplasty of the fingers and the restoration of the skin-fatty layer of the dorsal surface of the hand in boutonnière contracture]. In the period from 1983 to 1990 operations were conducted on 63 patients with postburn boutonnière contracture of 116 fingers. In 19 patients (42 fingers) tendoplasty of the extension apparatus of the fingers was carried out with simultaneous restoration of the dermatoadipose layer on the dorsal surface of the hand with an inguinal or dermatoadipose graft. In 39 patients the tendon of the extensor digitorum muscle was injured over several proximal interphalangeal articulations. A new method for treating boutonnière contracture is suggested. It is based on restoration of the continuity of the central fasciculus and the normal position of the lateral fasciculi with a single tendon autograft. The suggested method produced good results in 90% of cases, i.e. in all cases in which a sufficient volume of passive movements at the proximal interphalangeal articulation was restored in the preoperative period.


[The combined treatment of acute suppurative diseases of the fingers and hand using decamethoxin]. The authors discuss the results of complex treatment of 286 patients with acute pyoinflammatory diseases of the fingers and hand with the use of a new Soviet-produced antiseptic decametoxin. Panaris was diagnosed in 196 (68.5%), phlegmons and abscesses in 82 (29.7%), furuncle in 6 (2.1%) and carbuncle in 2 (0.7%) patients. 224 (78.4%) patients received out-patient and 62 (21.6%) in-patient treatment. The authors established that as the result of the applied complex treatment with the use of various antiseptic compositions containing decametoxin the mean duration of treatment was 7.8 days. The article discusses the causes of the disease, the methods of operative treatment, and management of patients in the postoperative period.


[The technic of esophageal extirpation with one-stage esophagoplasty using a stomach tube]. The article deals with the technical aspects of operation for extirpation of the esophagus through a cervico-abdominal approach in carcinoma (133 operations) and benign esophageal strictures (117 operations) with one-stage plastics by means of an isoperistaltic tube formed from the greater curvature of the stomach. The relative safety (2.8% of patients died) and high efficacy of the described operation allow it to be recommended for wide introduction into the practice of institutions engaged in surgery of the esophagus.


[An analysis of the reasons for relaparotomy in the surgical clinic]. The results of surgical treatment of 4,078 patients who underwent operation in 1985-1989 were studied. Due to the occurrence of complications in the early postoperative period, 182 relaparotomies were carried out in 164 patients. The main indications for relaparotomies were advanced underlying disease, surgical errors (tactical, technical), changes of the patients' immune status. Early diagnosis, timely operation with the use of technically correct manipulations, and the application of a full complex of intensive therapy measures in the postoperative period are important in the prevention of complications.


[A system for the clinico-immunological prognosis of infectious-inflammatory complications in planned surgery]. Postoperative infectious complications in planned surgery are a pressing problem. The preoperative condition of the patient and his immune system play an important role in their development. Fifty patients of the "risk" group were selected from the standpoint of classification of the etiological factors of secondary immunodeficiency states. Fifteen patients who had no symptoms classified as SIDS risk factors formed the control group. The immune status was examined before and on the third day after the operation in all patients of the main and control groups. Various infectious complications developed after the operation in 23 patients of the risk group. No such complications occurred in the control group. The preoperative immune status of the risk group patients differed significantly from that of the controls in 13 of 19 parameters. The patients of the main and control groups formed the instructing selection. An expert system of preoperative clinico-immunological prognostication of postoperative infectious-inflammatory complications was created on oasis of the methods of the theory of image recognition and instructing selection. The system was tested in a direct clinical experiment on 66 patients. The prognoses were correct in 89% of cases. It was thus proved that symptoms classified as SIDS risk factors influence the initial immunity status and the course of the postoperative period in planned surgery. An accurate, sensitive, and specific method for prognosticating postoperative infectious complications is suggested.


[Colored light stimuli in ERG for differential diagnosis of cone dystrophies]. We recorded electroretinograms (ERG) with white and color stimuli in normal persons and four patients with cone dystrophies. Kodak Wratten-filters in blue, blue-green, green, yellow and red were used for the color flashes. ERGs to all color stimuli were recorded at dark and light adapted conditions with different stimulus intensities, to 30 Hz flicker stimulation and with special filtering for oscillatory potentials. Selective blue cone-responses were obtained using strong blue flashes at a yellow background. Patients with cone dystrophies showed slightly to moderately reduced responses at dark adaptation to blue, blue-green, green and yellow stimuli. Red stimuli elicited only small responses with a markedly delayed b-wave implicit time. The light adapted recordings, flicker responses and oscillatory potentials were reduced to all color stimuli. However, differences between patients with cone dystrophies could be detected concerning the responses to red and green. In two patients the responses were reduced to the same degree to all color stimuli. Another patient had very small responses and no oscillatory potentials to red, but his responses were only moderately reduced to green. A patient with combined red-green cone and rod dystrophy had a blue cone hypersensitivity. Responses to blue and blue-green were large at all stimulus conditions, but responses to all other stimuli were much smaller. The blue cone ERG showed a prominent blue cone response. ERG recording to colored stimuli allows a separation of retinal dysfunction in patients with cone dystrophies.


[Multiple evanescent white dot syndrome]. A 38-year-old male patient experienced a unilateral visual acuity decrease to 20/60 and showed white dots at the level of the retinal pigment epithelial interface characteristic of multiple evanescent white dot-syndrome. Fluorescein angiography demonstrated early hyperfluorescent defects and some late staining. In spite of improvement of the visual acuity and the alterations of the fundus, an enlargement of the blind spot and some sharply demarcated depigmentations of the retinal pigment epithelium remain. This case shows, that already at the beginning of symptoms the characteristic white dots may be present. Enlargement of the blind spot and depigmentations of the retinal pigment epithelium may remain as defects after multiple evanescent white dot-syndrome.


[Detection of phosphoglucomutase in the cornea and corneal cell cultures using isoelectric focusing and enzymatic stain methods]. PGM-1 and its isoenzymes had been studied in the human cornea, in cornea cell cultures, in erythrocytes and lymphocytes. Qualitative and quantitative comparisons between cornea of healthy persons and cornea cell cultures of patients with cornea dystrophies showed differences between certain phenotypes and their signals of activity.


[Cardiac myxoma with central retinal artery occlusion, hemiparesis and epileptic seizure in a 9-year-old patient]. A cardiac myxoma is a very rare cause for a central retinal artery occlusion (CRAO). We report the case of a nine-year-old boy who had an epileptic seizure, the computer-tomographically suspected diagnosis was intracerebral granulomas of unknown etiology. Five months later he suffered an acute hemiparesis and a CRAO. Through echocardiography the diagnosis of a cardiac myxoma was determined. This case shows the importance of the differential diagnosis "cardiac myxoma" in cases of central retinal artery occlusion in young patients especially with additional neurological disorders.


[Reversible mydriasis and cycloplegia after isocyanate exposure]. We saw three patients working in the production of polyurethanes, who showed mydriasis and cycloplegia of unknown origin. In one patient there was also a transient opacification of corneal stroma. After discontinuation of working both mydriasis and cycloplegia resolved completely within 3-5 days. Because of the history and the analysis of chemical reactions involved in the production of polyurethanes there is strong evidence that both mydriasis and cycloplegia have been caused by topical and (/or) systemic intoxication with the aryl diet of the isocyanate molecule.


[Congenital facial hemiatrophy. Report of two pediatric cases]. In facial hemiatrophy two different types are known: the progressive acquired type, which is relatively more common, and the rare congenital non progressive form. Two children--both with congenital facial hemiatrophy--are presented, both showed a (horizontally and vertically) smaller lid fissure and antimongoloid configuration of the lid. In addition in one case we could observe: small corneal diameter, staphyloma posticum, paving stone degeneration of the retina, convergent strabismus, and vertical deviation, in the other case: ptosis, major astigmatism, and latent divergent strabismus.


[Endogenous candida endophthalmitis in a drug dependent patient: intravenous therapy with liposome encapsulated amphotericin B]. We report this exemplary case of a drug addict with candida endophthalmitis where we used the new form of amphotericin B, encapsulated in liposomes. We were able to reconfirm the reduced number of side effects and the minimized nephrotoxicity reported by authors of other specialties. In our patient, a reduction or elimination of the yeast was probably achieved, nevertheless, he developed a traction retinal detachment. In future cases of fungal endophthalmitis, we recommend liposomal amphotericin B in higher doses.


[Further development of a streak retinoscope with calibrated collimator]. Development and rationale of a new streak retinoscope have been discussed in an earlier paper. In contrast to classical retinoscopy this retinoscope determines the refraction by forming the narrowest streak in the test person's pupil with a calibrated collimator. In addition to the first prototype the extension of the lamp according to variation of the refraction is measured electronically and the data are transposed into a computer. This allows to freely choose a working distance in a range from about 30 to 120 cm and therefore it is possible to examine restless patients. Accuracy of the measurement was less than or equal to 0.25 D for Ametropia from -0.5 to 5.0 D in a working distance of 30 cm or from -4.0 to -1.5 D in 100 cm. In most persons it is not necessary to use additional glasses during refraction by choosing the adequate distance. Therefore this method can help to refract kids and other persons which could not be refracted by use of glasses. In addition this method excludes the accommodation of the patient as apposed to regular refractometers.


[Success of retinal surgery comparing phakic and pseudophakic eyes with reference to preoperative findings and the kind of lens implant]. We analyzed retrospectively 597 eyes over a minimum follow-up of 6 months and compared the results of pseudophakic eyes with phakic eyes. The repair of pseudophakic retinal detachment is more difficult than the surgery in aphakic retinal detachment. Pseudophakic retinal detachment had a more advanced retinal detachment and PVR-stages at the time of diagnosis. Localisation of the tear was more complicated and the surgery was more invasive than in phakic eyes. In 61.2% the repair was for pseudophakic eyes with posterior chamber lens 78%, with anterior chamber lens 81.8% and for iris-fixated IOL's 75%. Compared to this, the anatomic reattachment rate in phakic eyes was 94.8%. The pseudophakic group had less favorable visual results.


[Ocular findings in infection with Borrelia burgdorferi]. During the years 1988 to 1990 ten patients were observed in the eye clinic of RWTH Aachen with ocular findings of infection with Borrelia burgdorferi. 6 of them showed neuro-ophthalmological signs, 4 had uveitis. In both groups recent infections were documented in fresh and chronic stages of the disease, lasting longer than one year. The various clinical pictures of the disease following infection with Borrelia burgdorferi are described. As a vaccination against Borreliosis is not yet available, it is important to detect the antibodies against Borrelia burgdorferi early enough, to initiate an effective treatment.


[Errors of monocular localization in strabismic amblyopia. Two-dimensional distortion]. Spatial distortions, i.e. spatial localization errors, and precision of localization were measured monocularly in the central visual field of 7 normal observers, 17 strabismic amblyopes and 2 anisometropic amblyopes. The task of the subjects was to construct circles of 2 degrees, 4 degrees and 6 degrees radius around a fixation point, using the dominant and the amblyopic eye in turn. Normal observers set distances on the vertical meridian smaller than distances on the horizontal meridian. Anisometropic amblyopes showed localization errors and variances similar to those of normal observers. The amblyopic eyes of strabismic observers with a large angle strabismus and deep amblyopia showed significant, individually different localization errors correlating with the depth of amblyopia. Strabismics with microstrabismus exhibited parallels between monocular localization and dichoptic retinal correspondence.


[Objectified measurement of eye lens transparency in elderly probands. Results of a Scheimpflug photography study over the course of three and a half years]. Lens density has been followed-up in 32 healthy volunteers (mean age 65 years) over a period of 3.5 years. The negatives of Scheimpflug-photos (Topcon SL-45) were evaluated by linear microdensitometry in three measure planes (Joyce Loebl 3 CS). Correlation coefficients, curves of regression as well as calculations of the percentage of density change made obvious, that a minimal increase in lens density occurred in the anterior cortical region.


[Detoxication methods in pancreatogenic enzymatic peritonitis]. A therapeutic response was analyzed in 536 patients with acute destructive pancreatitis complicated by pancreatogenic enzymatic peritonitis. All the patients had the syndrome of endogenic intoxication and underwent intensive resuscitation with protease inhibitors, cytostatic agents, antibacterial, desensitization and detoxication treatment. If indicated, the patients with pronounced endogenic intoxication were subjected to forced diuresis, abdominal laparoscopic drainage and intraperitoneal dialysis, drainage of the thoracic lymphatic duct, hemosorption. Routine surgical intervention was performed. Up to 11% fell the general lethality due to a differentiated therapeutic approach.


[Reactive arthritis in young men in organized collectives]. The authors followed up 398 young males with reactive arthritis who had been admitted from organized collectives to hospital settings. Only one third of them could connect the development of reactive arthritis (ReA) with a survived infection: acute nasopharyngeal (26%), acute enterocolitis (7%), urethritis venerea (1%). Analysis of the clinical picture demonstrated that in case of indefinite ReA etiology together with the signs of myocardiac involvement one should not only exclude the presence of rheumatic attack but also to establish a possible relation of the disease with a prior survived poorly manifest intestinal infection. This fact should be taken into account if in the collectives, where from the patients were hospitalized, epidemic outbreaks of acute dysentery or cases of ReA and Reiter's disease etiologically connected with manifest intestinal infection were documented. Postenterocolitic origin of ReA could be suspected in patients with its continuous course and resistance to antiinflammatory therapy in whom arthralgia was preserved for a long time as was joint rigidity after a subsidence of joint changes due to the effect of acute inflammation. Besides, the detection of certain features of the articular syndrome and separate extraarticular signs characteristic of Reiter's disease and other conditions which comprised the group of seronegative HLA--B27-positive spondyloarthropathy are of high diagnostic value.


[Possibilities of the use of trypsinogen activation peptide in the pancreatic tissue in the diagnosis of pancreatic necrosis]. The test of peptide of trypsinogen activation (PTA) in various biological media is a principal new diagnostic test for acute pancreatitis. PTA levels in the pancreatic tissue and the plasma of the blood in acute experimental pancreatic necrosis reflects the process of the pathological activation of trypsin in the diseases focus and confirms the source of higher levels of activation peptide in the blood flow. Together with the values of other enzymatic systems of the pancreatic gland the discussed value permits one to make a more precise assessment of the mechanism and stages of autodigestive processes which develop in acute experimental pancreatitis.


[Clinico-morphological correlations of chronic pseudotumorous pancreatitis and cancer of the pancreas]. Examination of 57 pancreatic cancer patients and 40 subjects with chronic pseudotumorous pancreatitis revealed a resemblance in the clinical picture and uniform changes in the blood levels of pancreatic enzymes in both groups. Histological examination of 32 patients who expired from pancreatic cancer revealed total chronic pancreatitis manifested in inflammatory infiltration, focal destruction of the duct epithelium, atrophy, interstitial, periductular and perivascular sclerosis, dilatation of the ducts and pancreolithiasis. The character of the morphological changes in patients with pancreatic cancer corresponds to both primary and secondary origin of pancreatitis. Similarity of clinical manifestations and uniform changes in the blood content of pancreatic enzymes could be explained by the presence of chronic pancreatitis concurrent to pancreatic cancer.


[Indicators of lipid metabolism and lipid peroxidation system in patients with different types of lesions of the vessels of the lower limbs]. A total of 66 patients were investigated with IR imager and television capillaroscopy for the blood circulation in the vessels of low extremities as well as for the values of lipid metabolism and the system of lipid peroxidation. According to the status of the vascular system in the low extremities the examinees were divided into the groups with the normal status of the vascular system, the groups with the signs of venous insufficiency, microcirculatory disorders and atherosclerosis of major vessels. With the disorders of microcirculation in low extremities staged increments in the atherogenic shifts in the exchange were demonstrated. It was suggested that atherosclerotic changes in the arteries could be preceded with the hemodynamic changes in the venous and capillary systems due to rheological disorders due to the development of hypercoagulation which accompanied dyslipoproteinemias and other atherogenic shifts in the metabolism.


[Course and therapy of nonpancreatic complications of chronic pancreatitis]. Out of 647 patients with chronic recurrent pancreatitis followed up for 10-12 years 27 patients (4.2%) developed symptomatic gastroduodenal ulcers, 29 (4.5%) multiple gastroduodenal erosions. Ulcers and erosions emerged in patients with pronounced pancreatic bicarbonate insufficiency. Sucralfate treatment produced the best effect, while almagel plus vicalin were superior to gastrozepin. Relapses of ulcerogenesis were registered in 8 cases, multiple erosions in 11 cases, left pleural exudate in 8 cases in the presence of chronic pancreatitis exacerbation. Pancreatocardiac syndrome with cardialgias, a trend to arterial hypotonia, reduced voltage of ECG waves, occasional extrasystolic arrhythmia occurred in 45 patients (7%). It is shown that metabolic disorders of biogenic amines and lowered blood levels of insulin and C-peptide may underlie pathogenesis of pancreatocardiac syndrome.


[Ultrasonic methods of diagnosis of late complications after reconstructive surgery of the aorta and major arteries of the lower limbs]. Long-standing results were studied in 587 patients who sustained reconstructive vascular surgeries when aged under 16 years (the average age was 6 years and 4 months). Out of them 141 patients faced the necessity of repeated surgeries: in 115 (81.6%) patients the extremity function was preserved, in 19 (13.5%) a repeated amputation was performed, 7 patients (4.9%) expired. All those operated on had been actively followed up with the use of periodical instrumental investigation which included dopplerography and ultrasonic angiography. High significance of ultrasonic technique was proved as the use of it permitted one to detect late complications on the early stages of their development and perform surgical corrections before the development of life-threatening complications. Therefore repeated preventive surgeries were performed in 53 patients, 52 of them were a success. Repeated amputations were performed in 1 patient. All those operated on survived.


[Diffuse toxic goiter].

[Diffuse toxic goiter]. Diffuse toxic goiter is a common disease of the thyroid gland. As organospecific autoimmune condition the disease is not infrequently combined with endocrine ophthalmopathy and in this connection a more precise preparation of the patients to surgery is found to be mandatory. Elderly persons demonstrate atypical patterns of the disease course (insignificant enlargement of the gland, manifest depression, isolated cardial disorders without other symptoms of thyrotoxicosis, resistance to thyrostatic treatment). The patients need a life-long dispensary follow-up as diffuse toxic goiter is a condition of a relapsing type.


[Organization and methodology of the process of examinations in internal diseases in higher education]. The authors developed a system for examinations at the stage of postgraduate advanced training for therapists which included fundamental and practical classes. Fundamental classes are presented with certain tests with regard to the curriculum while the practical ones are based on the evaluation of professional clinical skill demonstrated at the examination of the patient. The developed system is realized with the help of computerized programmes CM-4. High objectivity and the wide spectrum of the programme evidence the practicability of the methods developed in the evaluation of knowledge in therapists.


Factors associated with intestinal amyloidosis in pigtailed macaques (Macaca nemestrina). Factors associated with intestinal amyloidosis in pigtailed macaques (Macaca nemestrina) were studied in 74 cases at the Washington Regional Primate Research Center. The medical records of monkeys during the 5-year period from 1983 to 1988 were analyzed to determine the age at death, age at first episode of diarrhea, number of episodes of diarrhea, episode and cumulative duration of diarrhea, and etiologies of diarrhea. Univariate analysis, using one control for each case, indicated that only episode duration was related to intestinal amyloidosis. Affected monkeys had significantly longer mean episode durations of diarrhea. None of the etiologies examined--bacteria, protozoa, fungi, and simian retrovirus--were significant risk factors for amyloid deposition in the intestinal tract.


Diarrhea rates and risk factors for developing chronic diarrhea in infant and juvenile rhesus monkeys. Ten independent risk factors were evaluated in an effort to identify predictors of problem diarrhea at weaning and chronic diarrhea in infant and juvenile rhesus monkeys (Macaca mulatta) at the California Primate Research Center. None of the variables proved to be a significant predictor of problem diarrhea at weaning; however, two of the variables were significant predictors for developing chronic diarrhea. Odds ratios, adjusted for other variables in the logistic regression model, showed that compared with females, males were nearly three times more likely to develop chronic diarrhea, and nursery-reared animals were 7.5 times more likely to develop chronic diarrhea than were breast-fed animals. The annual incidence rates for problem diarrhea at weaning for 1978, 1979, and 1980 were 49%, 37%, and 41%, respectively. A weighted average annual incidence rate for problem diarrhea at weaning for the 3-year period was 39%. The incidence rate for chronic diarrhea for the 3-year period was 49%.


Endometrial venous aneurysms in three New Zealand white rabbits. Hematuria in rabbits has been associated with uterine adenocarcinoma, uterine polyps, renal infarction, urolithiasis, cystitis, bladder polyps, and pyelonephritis. Three adult female New Zealand White rabbits (Oryctolagus cuniculus) developed apparent hematuria, as suggested by blood in their excreta pans. They had been immunized with antigen-adjuvant emulsions, but had uneventful clinical histories. Physical examination disclosed no abnormalities, and laboratory tests, including hematology, serum chemistries, urinalyses, urine cultures, ultrasonography, and intravenous pyelography disclosed mild anemia, hematuria, and proteinuria in two of the rabbits. Antibiotic therapy failed to alleviate clinical signs. Two rabbits were euthanized because of persistent urogenital bleeding and the third rabbit underwent exploratory laparotomy and ovariohysterectomy. Multiple endometrial venous aneurysms were present in the uteri of all rabbits and urogenital bleeding was attributed to episodic bleeding from these lesions. Varices and aneurysms of uterine subserosal and myometrial venous plexuses, but not of endometrial vessels in women have been reported. To our knowledge, endometrial venous aneurysms have not been reported in animals previously. Our findings indicate that the differential diagnoses for sporadic apparent hematuria in female rabbits should include endometrial aneurysms.


Effects of sex hormones on fulminant hepatitis in LEC rats: a model of Wilson's disease. LEC rats, which have hereditary hepatitis and have recently been proposed as an animal model for Wilson's disease, were examined to determine the effects of sex hormones on fulminant hepatitis. After the rats had undergone ovariectomies or orchidectomies (castration) and were compared with intact rats, the age at the onset of fulminant hepatitis was not substantially altered but the survival rates decreased from 50% to 12.5% for females and 75% to 14.3% for males, indicating that sex hormones did not influence the occurrence of fulminant hepatitis but influenced mortality due to fulminant hepatitis. When testosterone was administered to the ovariectomized or orchidectomized rats, the survival rate increased to over 90% in both sexes. In contrast, estradiol did not affect the survival rate of either sex but affected the onset of fulminant hepatitis. That is, with the administration of estradiol, the age at which serum GPT activity reached its maximum was delayed 4 weeks in ovariectomized rats and 6 weeks in orchidectomized rats as compared with intact rats. A similar but somewhat weaker tendency appeared in rats given progesterone. The results of our study indicate that sex hormones have no effect on the rate of occurrence of hepatitis but affect the progression of hepatitis. In particular, testosterone increased the survival rate of rats with fulminant hepatitis, and exogenous estradiol delayed the onset of hepatitis for several weeks.


A reproducible method for producing and quantifying the stages of fracture repair. Male CD-1 mice, 4 to 6 months of age, were used to establish a reproducible model to study the stages of fracture repair. A custom-designed fracture apparatus was constructed, and trials with it demonstrated its capacity to reliably reproduce a closed fracture of the tibia. Dietary and sleep habits in the treated mice were the same as unfractured control mice. Four stages of fracture repair were documented and the duration of each stage was quantifiable and reproducible. The last stage of fracture repair was completed by 21 days postfracture. The reproducibility of the fracture, the reproducibility of the times and stages of fracture repair, the relatively short time to complete the fracture repair process, and the minimal discomfort which allowed the mice to maintain a normal daily routine, suggest that this is an ideal animal model for studying the fracture repair process.


Lymphoblastic lymphoma in a colony of N:NIH(S)-bg-nu-xid mice. During a 1-year period, 28 animals from a breeding colony of N:NIH(S)-bg-nu-xid mice were discovered to have rapidly enlarging subcutaneous swellings in the ventral, cervical, and axillary regions. Five of the mice also had hind limb paresis. Twenty-two of the mice were heterozygous nude females, five were homozygous nude males, and one was a homozygous nude female. All of the above mice were homo- or hemizygous for the beige and X-linked immunodeficiency mutations. The average age of the mice was 8.3 months. Generalized enlargement of the peripheral and internal lymph nodes was present at the time of necropsy examination. Other lesions commonly noted at necropsy included splenomegaly (15 mice), pale and thickened ventral lumbar spinal musculature (11 mice), and opaque, thickened meninges of the brain (10 mice). Histologic examination consistently disclosed infiltrates of neoplastic lymphoblasts in multiple tissues including lymph nodes, spleen, bone marrow, and meninges of the brain and spinal cord. The cells were positive for IgG on immunofluorescent staining, suggesting that the tumors were of B cell origin. The neoplasms observed in these mice have several similarities to tumors found in immunodeficient humans, suggesting that these mice may serve as useful animal models of lymphoma.


Convulsions in senescence-accelerated mice (SAM-R/1/Eis). Senescence-accelerated mice (SAM) are one of the animal models used for studying senescence, which consist of several substrains such as SAM-R/1, R/2, P/1, P/2. SAM-R/1/Eis maintained in Eisai Tsukuba Research Laboratories, Ibaraki, Japan, was originally introduced as a substrain of a normal control SAM-R/1 from Kyoto University, Japan. We have noted signs of convulsions in SAM-R/1/Eis mice during routine animal care, particularly while changing cages. We identified the clinical signs and determined the concentrations of glucose and immunoreactive insulin in plasma of SAM-R/1/Eis mice. There were no differences in the male:female ratios of mice showing prodrome only, grand mal, or no-signs. The ages at which prodrome and grand mal were first noted peaked between 20 and 25 weeks. Concentrations of glucose and immunoreactive insulin in plasma did not indicate the mice were in insulin hypoglycemia, which is one cause of convulsions. AKR strain mice, some of which originated with the SAM strain are known to become convulsive by repeated "throwing" stimulations. Conversely, in SAM-R/1/Eis, throwing stimuli are not needed to cause convulsive signs. Thus it is likely that in SAM-R/1/Eis mice the signs are triggered by repeating mild environmental changes, such as changing cages. The results of this study show that SAM-R/1/Eis is neither a normal control strain, nor an original SAM-R/1 strain. But it is possible that SAM-R/1/Eis is another useful animal model for studying spontaneous convulsion.


Persistent free-running circannual reproductive cycles during prolonged exposure to a constant 12L:12D photoperiod in laboratory woodchucks (Marmota monax). Serum levels of gonadal steroid were assayed at approximately 3-month intervals in groups of 5 to 8 male or female woodchucks which were exposed to a natural photoperiod for 1 year as yearlings or 3 years as adults (Study 1), or a constant photoperiod of 12L:12D from birth for 4.5 years (Study 2). After 4.5 years of 12L:12D, food intake was measured in November and compared with that in natural photoperiod animals (Study 3). Other groups of 11 males and 3 females were housed in 12L:12D for 2.5 years after capture at 2 months of age, and gonadal structure and serum steroid levels in November were compared with those of animals at selected times in the normal annual cycle (Study 4). All animals were provided food and water ad libitum and were not induced to hibernate. In Study 1, normal circannual breeding season elevations in testosterone in males and in progesterone in females were detected in most animals maintained in natural photoperiod. In Study 2, similar cycles persisted for 4.5 years in animals exposed to 12L:12D. However, based on quarterly blood samples, obvious asynchrony relative to natural light animals appeared to develop after 2, 3, or 4 years, with apparent free-running intervals of about 10 to 11 months. In Study 3, mean daily food consumption in late autumn for woodchucks in the 12L:12D group was 72% greater than animals in the natural photoperiod. In Study 4, some woodchucks exposed to 12L:12D for only 2.5 years had prematurely increased spermatogenic activity, Leydig tissue development, and elevated serum testosterone levels in November. They were similar in November to those in natural photoperiod animals in March, and significantly greater than those in natural photoperiod animals in November when normal regression and repair of the testis was complete. Likewise, females in the 12L:12D group had luteinized follicles and elevated progesterone in November which were not noted in natural photoperiod animals and which were similar to those observed during the spring in unbred females under normal conditions. The results suggest that circannual cycles of metabolic and reproductive activity in woodchucks persist in the absence of normal changes in photoperiod, are entrained to seasonal changes in the natural photoperiod, and can recede to a periodicity of less than 12 months within 2.5 to 4 years of laboratory maintenance in 12L:12D.


Pickle barrels as enrichment objects for rhesus macaques. Two breeding groups of rhesus monkeys (Macaca mulatta) housed in outdoor enclosures on Key Lois island were observed for 84 hours. Instantaneous scan sampling of a focal animal was used to gather data to test hypotheses concerning frequencies of agonistic and affiliative behaviors as well as differential use of pickle barrels as enrichment objects. Type of barrel used, behavior, and age/sex class of the animal were noted. Barrels were arranged three ways: unattached, on a swivel, and stationary. The behaviors of animals in an enriched environment were compared with control condition animals, which did not have pickle barrels. Animals in an enriched environment accounted for 60.8% (n = 56) of total affiliative contact, 62.2% (n = 399) of total social grooming, and 26% (n = 5) of total agonistic noncontact. A total of 134 scans of barrel use were observed. Analyses of the data showed that swivel and stationary barrels were used the most (55% of all scans of barrel use). Yearlings, juvenile females, and old males used barrels most often (82.8% of all scans of barrel use), although they constituted only 39% of the enriched environment population. In this study, pickle barrels provided enrichment for young and old animals of both sexes.


Ultrasound-guided blood sampling of rabbit fetuses. A rabbit animal model for hemolytic disease of the newborn has been previously described. However, evaluating the effects of this disease was limited to histologic and hematologic examinations of liveborn kitlings. To assess the feasibility of in utero blood sampling, we performed ultrasound-guided cardiac sampling of 50 fetuses in 16 New Zealand White does on days 26 and 27 of gestation. The overall rate of successful sampling was 80%. The procedure-related mortality declined to 35% by the third phase of the study. The mean (+/- SD) hematocrit (%) and reticulocyte values (#/100 RBCs) on day 26 were 26.3 +/- 3.3 and 35.6 +/- 5.1, respectively; values on day 27 were 31.3 +/- 4.9 and 27.5 +/- 7.6. The results of this study suggest that hematologic data can be obtained from rabbit fetuses in the majority of cases with only moderate fetal loss.


Evaluation of subcutaneous chambers as an alternative to conventional methods of antibody production in chickens. We compared antibody levels among serum, egg yolk extract, and granuloma fluid in chickens immunized with bovine serum albumin (BSA). One group of hens was immunized by intramuscular and subcutaneous injection of bovine serum albumin in complete Freund's adjuvant, followed by two subsequent booster injections in incomplete Freund's adjuvant. Two other groups were surgically implanted with plastic, perforated wiffle balls (subcutaneous chambers). After a 30-day recovery period, one of the groups with subcutaneous chambers was immunized with BSA in sterile water with two subsequent boosts. The other group was injected with only sterile water. Serum samples, eggs, and granuloma fluid were collected biweekly and analyzed to determine specific IgG, total IgG, and total protein. The subcutaneous chambers were well tolerated. Quantitative ELISAs of serum, egg yolk extract, and granuloma fluid specimens disclosed that specific antibody levels were present in all specimens by 2 weeks after primary immunization. During the course of the experiment, specific antibody levels of serum and egg yolk specimens were significantly higher than those of granuloma fluid (P less than 0.05). However, an additional injection of antigen into the subcutaneous chambers resulted in specific antibody levels in granuloma fluid specimens that were comparable to those of serum and egg yolk extract. Use of subcutaneous chambers in chickens may be a viable alternative to routine antibody production methods.


An occipito-temporal syndrome in adolescents with optimally controlled hyperphenylalaninaemia. The study included 16 adolescents with optimally controlled hyperphenylalaninaemia (McKusick 26160), of whom six did not require treatment according to conventional criteria. All except the two patients with lowest median serum phenylalanine level throughout childhood (most values at 200-300 mumol/L) had white matter abnormalities detectable with magnetic resonance imaging. The lesions were particularly prominent in the watershed regions between the posterior and middle cerebral arteries. In most patients with moderate or severe hyperphenylalaninaemia frontal white matter lesions were present as well. Normal proton magnetic resonance spectra indicated that the lesions were stable. Occipital EEG abnormalities were frequent, and deficient performance on a pattern-recognition test was a characteristic neuropsychological finding. Serum phenylalanine levels at about 300 mumol/L or below throughout childhood and early adolescence may be required to avoid lesions. The present study demonstrates the limitations of even an optimally controlled dietary regimen in hyperphenylalaninaemia.


Allopurinol challenge test in children.

Allopurinol challenge test in children. The allopurinol challenge test was performed on 44 healthy subjects (28 children and 16 adolescents) in order to establish normal values of urinary orotic acid excretion following allopurinol ingestion in the paediatric population. The subjects were divided into three groups according to their age: 6 months to 6 years; 6 years to 10 years; and 10 years to 17 years. They were given 100 mg, 200 mg, or 300 mg of allopurinol, respectively (based on age) in a single oral dose. Maximum peak urinary orotic acid levels following ingestion of allopurinol were 13.0 (n = 14), 9.3 (n = 14), and 10.2 (n = 16) mumol/mmol creatinine in the three groups, respectively. In all children tested the peak orotic acid level was 3.1 +/- 2.7 mumol/mmol creatinine (mean +/- SD, n = 44). This allopurinol challenge test was also performed in six children with urea-cycle disorders, including five females with ornithine transcarbamylase (OTC) deficiency, all of whom demonstrated abnormally elevated levels of urinary orotic acid (peak levels of 26-134 mumol/mmol creatinine) following allopurinol ingestion.


Prenatal diagnosis of non-ketotic hyperglycinaemia. Non-ketotic hyperglycinaemia (NKH) is a devastating neurological disease for which there is no effective therapy. Consequently, most couples with a pregnancy known to be at risk for NKH request prenatal diagnosis. We have applied the combination of chorionic villus (CVS) assay for glycine cleavage enzyme activity and determination of amniotic fluid glycine concentration to increase the reliability of prenatal diagnosis for this disorder beyond that of each of these methods alone. All 15 of the at-risk pregnancies monitored had CVS glycine cleavage assay and five also had amniotic fluid glycine measurements. Two cases had no detectable cleavage activity in CVS and one gave uninterpretable enzyme results. Amniotic fluid glycine concentration was increased in all three and NKH was confirmed by abortus tissue assays for cleavage activity and amino acids. The remaining 12 case had activity in CVS (two also had normal amniotic fluid glycine levels) and delivered unaffected infants. Four of these 12 cases had cleavage activities below or at the low end of the normal range, perhaps indicating carrier status. We believe that the combination of CVS glycine cleavage assay and amniotic fluid glycine measurement is currently the best approach to the prenatal diagnosis of NKH.


Fructose and glucagon loading in siblings with fructose-1,6-diphosphatase deficiency in fed state. Hypoglycaemia induced by fructose administration is one of the diagnostic clues to fructose-1,6-diphosphatase (FDPase) deficiency (McKusick 229700). However, the pathological mechanism of this reactive hypoglycaemia is not fully known. This paper describes two siblings with FDPase deficiency, diagnosed enzymatically in leukocytes, who failed to correct reactive hypoglycaemia after glucagon administration even in the fed state, supporting a possibility that disturbed hepatic phosphorylase activity may be a main cause of reactive hypoglycaemia.


In vivo 13C-NMR evaluation of glycogen content in a patient with glycogen storage disease. Glycogen storage disease was suspected in a 10-month-old boy. Initial technical problems did not permit the determination of the precise enzyme, deficiency, and type VI glycogen storage disease was only diagnosed at the age of 2 years. In the mean time, natural abundance 13C nuclear magnetic resonance evaluation of muscular and hepatic glycogen content indicated normal muscular glycogen and increased hepatic glycogen in our patient, a finding which strongly argued for the diagnosis of type VI glycogen storage disease. Even though the use of nuclear magnetic resonance might seem, in this situation, a somewhat circuitous means of reaching the diagnosis, it appears that nuclear magnetic resonance could provide a useful tool for a non-invasive diagnosis of glycogen storage diseases.


Measurement of acyl-CoA dehydrogenase activity in cultured skin fibroblasts and blood platelets. The measurement of acyl-CoA dehydrogenase activity is an essential part of the investigation of patients with suspected defects of fatty acid oxidation, and recently the organometallic oxidant ferricenium hexafluorophosphate has been introduced as an electron acceptor for these assays. However, we show that when medium-chain acyl-CoA dehydrogenase activity was measured in cultured skin fibroblasts and platelets from patients with proven defects of this enzyme, there was considerable residual enzyme activity when this electron acceptor was used. The ferricenium assay is not as specific as the anaerobic ETF-linked assay in the biochemical diagnosis of medium-chain acyl-CoA dehydrogenase deficiency in fibroblasts, and therefore is of limited clinical applicability in its present form.


Carrier detection in glutaric aciduria type I using interleukin-2-dependent cultured lymphocytes. Cultured interleukin 2 (IL-2)-dependent leukocytes from 13 patients with glutaric aciduria type I, 12 obligate carriers, 105 family members and 31 normal controls were assayed for glutaryl-CoA dehydrogenase activity. Of the 13 affected patients, 10 (all Ojibway Indian) had residual enzyme activity (2-13% of control) and 3 patients (all non-Indian) had undetectable enzyme activity. There was partial overlap between the distribution of enzyme activity in obligate heterozygotes and in normal controls (mean values +/- SD: 6.29 +/- 0.94 and 10.75 +/- 2.58 nmol/h per mg protein respectively). Using an arbitrary cutoff level of < 7 nmol/h per mg protein as presumptive evidence of carrier status, the observed frequency of carriers did not differ significantly from that expected from their a priori risk of carrier status. Thirteen per cent of the family members had inconclusive status (activity between 7 and 8.5 nmol/h per mg protein). The method appears suitable for carrier detection, although definitive carrier assignment awaits identification of the mutation(s) responsible for glutaric aciduria type I.


Characterization of phosphofructokinase-deficient canine erythrocytes. Dogs homozygously affected with muscle-type phosphofructokinase (PFK) deficiency had about 20% of normal erythrocyte PFK activity and exhibited a compensated haemolytic anaemia. Erythrocyte glucose-6-phosphate and fructose-6-phosphate concentrations were increased and dihydroxyacetone phosphate and 2,3-bisphosphoglycerate values were below normal in affected dogs. Other intermediates distal to the PFK step were not significantly below normal and fructose-1,6-bisphosphate was even above normal. Erythrocyte ATP was higher than normal in affected dogs owing to the reticulocytes present. Abnormal adenylate metabolism was demonstrated by low ATP/AMP and ADP/AMP ratios and the inability to maintain ATP content when affected erythrocytes were incubated with cyanide. Glucose-1,6-bisphosphate content was normal, and fructose-2,6-bisphosphate content in affected canine erythrocytes was higher than normal. Studies of erythrocyte PFK isozymes revealed altered enzyme kinetic properties in affected dogs which appeared to be due to the loss of the M-type subunit.


N-acetylglucosamine 6-sulphatase deficiency in a Nubian goat: a model of Sanfilippo syndrome type D (mucopolysaccharidosis IIID). A male Nubian goat (SD-1) presented at birth with neurological manifestations consistent with a lysosomal storage disease. Histological studies of tissue obtained at autopsy suggested glycosaminoglycan storage. Total urinary glycosaminoglycan levels, as measured by the uronic acid method, were elevated but overlapped with levels in a younger control goat. However, N-sulphate content was increased 2- to 5-fold, suggestive of heparan sulphate excretion, and this elevation was confirmed by cellulose acetate electrophoresis. Further, urinary levels of free N-acetylglucosamine 6-sulphate were increased 6-fold over controls, SD-1 cultured skin fibroblasts, labelled with [35S]sulphate from the incorporated twice as much radioactivity into macromolecular material as did normal fibroblasts. Forty-eight hours after removal of [35S]sulphate from the medium the SD-1 fibroblasts retained 58% of the label, whereas in control fibroblasts it had declined to 20%, indicative of [35S]proteoglycan storage in SD-1. The assay of fibroblast extracts revealed a profound deficiency of N-acetylglucosamine 6-sulphatase whereas eight other activities including beta-mannosidase, arylsulphatase B, iduronate 2-sulphatase, N-acetylgalactosamine 6-sulphatase, and heparin sulphamidase were normal. Mixing of SD-1 sonicates with normal sonicates showed no evidence of an inhibitor, and mixing of SD-1 sonicates with Sanfilippo D cell sonicates yielded no activity. These data ruled out multiple sulphatase deficiency and suggested the first example of the human Sanfilippo syndrome, type D (N-acetylglucosamine 6-sulphatase deficiency) in goats.


Serum selenium levels in individuals on PKU diets. Serum selenium levels in 73 patients with phenylketonuria were significantly lower than in controls. The phenylketonuric and hyperphenylalaninaemic individuals taking the non-supplemented amino acid mixture generally had lower levels: 36% were below the normal range as defined by our laboratory, compared with 19% in the supplemented group. The low levels were present even in those on diet, who had a greater phenylalanine tolerance--that is, a tolerance for more than 9 x 50 mg phenylalanine exchanges per day, in other words a higher intake of natural protein. Individuals on long-term synthetic diets may be at risk for selenium deficiency even on selenium supplements. In areas where the soil may be low in selenium, the deficiency may be aggravated. Long-term low levels may impair health but the required amount of selenium supplementation remains uncertain.


Elevated plasma carnitine in the hepatic form of carnitine palmitoyltransferase-1 deficiency. In a boy with a defect in fatty acid oxidation due to the hepatic form of carnitine palmitoyltransferase-1 deficiency, plasma carnitine concentrations were found to be twice normal. The elevation in plasma carnitine levels was accompanied by an unusually high renal threshold for free carnitine, suggesting a secondary increase in carnitine transport. Similar to other fatty acid oxidation disorders involving the carnitine cycle, urinary dicarboxylic acids were not abnormally elevated during illnesses. The combination of elevated plasma carnitine levels and absence of dicarboxylic aciduria may help to distinguish the hepatic form of carnitine palmitoyltransferase-1 deficiency from other defects in fatty acid oxidation.


Topographic dissociation between mitochondrial dysfunction and cell death during low-flow hypoxia in perfused rat liver. Although it has been postulated that mitochondrial dysfunction and oxygen radicals may play a role in the mechanisms of hypoxic liver injury, little is known about the intralobular topographic relationship between a disorder in oxygen metabolism and cell death during hypoxia. Intralobular heterogeneity of mitochondrial dysfunction and its topographic relationship to cell death during 25% low-flow hypoxia was visually demonstrated in perfused rat hepatic microcirculatory units by dual-color digital microfluorography using rhodamine 123, a fluorochrome sensitive to the mitochondrial membrane potential, and propidium iodide, which labels the nuclei of nonviable cells with a concomitant enhancement of fluorescence. In the normoperfused liver from fasted rats, no significant depletion of the mitochondrial membrane potential values was observed in any portion of lobules during 100 minutes. In the liver treated with the 40-minute hypoxia, the mitochondrial membrane potential values within the entire lobule were significantly decreased. After a longer period of hypoxia, the pericentral fluorescence decreased to background levels, and the intralobular gradient showed a steep decline midway between the periportal and pericentral regions. Cell death was at first prominent in the midzone at 40 minutes, and then extended towards the pericentral regions, forming centrilobular necrosis at 100 minutes. Although allopurinol diminished the early midzonal cell death and retarded the development of centrilobular necrosis, it did not attenuate depression of the mitochondrial membrane potential. It is conceivable that mitochondrial dysfunction per se may not be related to the intralobular distribution of cell death during hypoxia. On the other hand, mitochondrial dysfunction in the centrilobular regions seems to be an important event leading to early midzonal oxidative tissue damage in that this change may involve ATP breakdown and the subsequent enhancement of xanthine oxidase-mediated oxidative cell injury in the midzonal regions, which are incompletely oxygenated.


Regulation of p26-bcl-2 protein levels in human peripheral blood lymphocytes. The protein encoded by the bcl-2 (B cell lymphoma/leukemia-2) proto-oncogene has been implicated in the regulation of lymphocyte cell survival and has been shown to interfere with programmed cell death (apoptosis). Recently, controversy has emerged regarding the expression of bcl-2 in circulating peripheral blood lymphocytes (PBL) and its correlation with lymphocyte proliferation. Using immunohistochemical methods, Pezzella et al. (Pezzella F, Tse G, Cordell J, Pulford K, Gatter K, Mason D. Am J Pathol 1990; 137:225-32) detected abundant amounts of Bcl-2 protein in resting PBLs and observed an inverse correlation between bcl-2 expression and cellular proliferation in lymphoid tissues. In contrast, previous in vitro studies of bcl-2 mRNAs showed that expression of this gene is very low in unstimulated PBLs but can be markedly induced when lymphocytes are stimulated to proliferate in culture (Reed JC, Tsujimoto Y, Alpers JD, Croce CM, Nowell PC. Science 1987; 236:1295-9; Granniger W, Seto M, Boutain B, Goldman P, Korsmeyer S. J Clin Invest 1987; 80:1512-5). To resolve this issue, the regulation of p26-Bcl-2 protein levels was examined in freshly isolated and in cultured human PBLs by immunoblotting using two different polyclonal antisera specific for the human Bcl-2 protein. When freshly isolated from whole blood, resting PBLs contained easily detectable amounts of p26-Bcl-2 protein that did not significantly change in culture when cellular proliferation was stimulated with mitogenic lectins and lymphokines. If processing of the blood was delayed, however, p26-Bcl-2 protein was low or undetectable in resting PBLs and underwent marked increases in its relative levels after in vitro stimulation of PBLs by mitogenic lectins and lymphokines. The findings indicate that bcl-2 is normally expressed in quiescent circulating lymphocytes, consistent with a role for this gene in the maintenance of lymphocyte survival, and illustrate the importance of correlating in vitro and in vivo expression data.


Chinese hamster ovarian cells transfected with human parathyroid hormone-related protein cDNA cause hypercalcemia in nude mice. Parathyroid hormone-related protein (PTH-rP) has been implicated as a causative factor in the humoral hypercalcemia of malignancy. Animal models of hypercalcemia of malignancy that have traditionally utilized human or animal tumors or injections or infusions of hypercalcemic factors have limitations that may prevent exact delineation of the biologic effects of tumor-produced PTH-rP. To assess the effects of tumor-produced PTH-rP in vivo, we have transfected Chinese hamster ovarian (CHO) cells with cDNA encoding human preproPTH-rP-(1-141) which then stably express only PTH-rP. We inoculated these tumor cells into nude mice, and compared tumor-bearing nude mice with similar tumor-bearing nude mice inoculated with non-transfected CHO cells using standard parameters of calcium homeostasis. The nude mice carrying tumors expressing PTH-rP became hypercalcemic over 12 to 18 days. Blood ionized calcium values correlated positively with plasma PTH-rP concentration and tumor volume. Plasma PTH-rP concentrations in hypercalcemic mice were similar to those reported in humans with hypercalcemia of malignancy. Bone histology and histomorphometry from hypercalcemic nude mice demonstrated increased bone resorption when compared with mice bearing nontransfected CHO tumors. We have produced an animal model of tumor-produced PTH-rP by transfecting CHO cells with the cDNA for PTH-rP and inoculating these tumor cells into nude mice. Hypercalcemia in this model is mediated in part by the effects of PTH-rP to increase osteoclastic bone resorption. The model is advantageous because PTH-rP alone is secreted in a prolonged, constitutive fashion with pharmacokinetics similar to naturally occurring tumors. It should prove to be a useful model to study the effects of tumor-produced PTH-rP in vivo.


Carbohydrate metabolism in human renal clear cell carcinomas. Renal cell carcinomas can be subclassified into clear cell carcinomas, chromophobe cell carcinomas, chromophilic cell carcinomas, and oncocytomas. Previous studies, in which no distinction among the different types of renal cell tumors and their grades of malignancy was performed, showed that these tumors had high glycolytic rates. The carbohydrate metabolism of control human kidney samples and renal clear cell carcinomas with different degrees of cytologic malignancy (G I, G II, and G III) was studied by determining the glycogen and glucose-6-phosphate levels and the activities of key enzymes involved in glycolysis (hexokinase, glucokinase, pyruvate kinase), gluconeogenesis (glucose-6-phosphatase, fructose-1,6-diphosphatase), and the pentose phosphate pathway (glucose-6-phosphate dehydrogenase) in these tissues and compared with those of a limited number of chromophilic cell carcinomas, chromophobe cell carcinomas, and oncocytomas. The glycogen and glucose-6-phosphate levels were significantly higher in G I, G II, and G III clear cut carcinomas than in control kidneys; glucokinase, hexokinase, and glucose-6-phosphate dehydrogenase activities remained unchanged, pyruvate kinase activity was enhanced, and glucose-6-phosphatase as well as fructose-1,6-diphosphatase activities were strongly reduced when compared with control kidney values. In chromophilic cell carcinomas glycogen content, glucose-6-phosphate dehydrogenase, and pyruvate kinase activities were elevated, while fructose-1,6-diphosphatase activity was reduced. In chromophobe cell carcinomas glycogen content was elevated and gluconeogenesis was reduced, whereas glycolysis was not activated. In oncocytomas glycogen was not detected and glucose-6-phosphate dehydrogenase, pyruvate kinase, and fructose-1,6-diphosphatase activities remained unchanged. It has been demonstrated that a series of characteristic changes occur in the carbohydrate metabolism of renal clear cell carcinomas: glycogen and glucose-6-phosphate levels increase, glycolysis is activated, and gluconeogenesis is reduced. Furthermore, the alterations of the carbohydrate metabolism within clear cell carcinomas are clearly distinct from those observed in chromophilic cell carcinomas, chromophobe cell carcinomas, and oncocytomas.


Platelet-derived growth factor receptor-beta is induced during tumor development and upregulated during tumor progression in endothelial cells in human gliomas. Endothelial cells proliferate during brain development, are quiescent in normal adult brain but proliferate again under pathologic conditions such as glioma growth. The vascular phenotype of low grade glioma is comparable to normal brain, however high grade gliomas are focally highly vascularized and there is associated prominent endothelial cell proliferation. The mechanisms of this change in vascular phenotype are unknown but there is evidence that growth factors play an important role in this process as well as in normal angiogenesis and vascular differentiation. To investigate whether endothelial cells become activated during tumorigenesis and progression of human gliomas by a platelet-derived growth factor (PDGF) dependent pathway, we analyzed platelet-derived growth factor receptor-beta (PDGFR-beta) expression by in situ hybridization and immunocytochemistry in normal human brain, astrocytoma (grade II), anaplastic oligo-astrocytoma (grade III), and glioblastoma multiforme (grade IV). PDGFR-beta mRNA was not detectable in the vessels of normal human brain, but was expressed in the vasculature of low and high grade gliomas, particularly in endothelial cell proliferations in glioblastomas. The expression of the receptor in the tumor microvessels, was confirmed by double immunofluorescence in which the staining appeared to be in the endothelial cells. Primary cultures of endothelial cells derived from glioblastoma multiforme maintained receptor expression for 2 days in vitro, whereas it was not detectable in vitro in endothelial cells derived from normal brain. Tumor cells in all grades of glioma expressed very little PDGFR-beta mRNA in situ. Our results indicate that the malignant phenotype in human glial tumors is associated with an upregulation of the PDGFR-beta on endothelial cells of vessels which vascularize the tumor. These findings may contribute to our understanding of the mechanisms that regulate vessel growth and differentiation in normal and pathologic states.


Relationship of cytochrome P-450 activity to Clara cell cytotoxicity. III. Morphometric comparison of changes in the epithelial populations of terminal bronchioles and lobar bronchi in mice, hamsters, and rats after parenteral administration of naphthalene. The purpose of this study was to define, in quantitative terms, epithelial alterations produced by the cytochrome P-450-activated Clara cell cytotoxicant, naphthalene, in lobar bronchi and terminal bronchioles of three species with differing sensitivity: mouse, rat, and hamster. Adult mice, hamsters, and rats were treated intraperitoneally with a single dose of naphthalene ranging from 0 mg/kg up to the approximate LD50. The animals were killed 24 hours postinjection and the changes in airway epithelium characterized by light microscopic morphometry. In mouse, bronchiolar epithelial thickness was significantly elevated by low, but not high, doses; ciliated cell number increased and Clara cell number decreased in a dose-dependent fashion. Vacuolated Clara cell number increased in all treated mice. In rat and hamster, bronchiolar epithelial thickness or cell number did not change. In mice, bronchial epithelial thickness was unchanged except at high doses, but both ciliated and Clara cell number was decreased. In bronchi of rats, epithelial thickness and numbers of nonciliated, ciliated, and basal cells were unchanged. In bronchi of hamsters, both ciliated and nonciliated cell number were decreased. (a) In mice, naphthalene-induced acute bronchiolar toxicity involves not only Clara cells, but also affects the purported nontarget cell type (ciliated cells). (b) In rats and hamsters, bronchiolar epithelium is insensitive to naphthalene injury. (c) In mice, injury to bronchi occurs at higher doses than in bronchioles and involves both ciliated and nonciliated cells. (d) In rats, bronchi are insensitive. (e) In hamsters, bronchi are more sensitive than bronchioles. This study emphasizes the variability of response by species, airway and epithelial cell type to cytochrome P-450-mediated pulmonary toxicants and the need for precise quantitative methods of defining both cytotoxic and metabolic events.


Thrombospondin induces glomerular mesangial cell adhesion and migration. Extracellular matrix components are known to modulate mesangial cell functions as adhesion, motility and proliferation. Among other extracellular matrix components, mesangial cells have been recently described to secrete thrombospondin (TSP), a high molecular weight glycoprotein, produced by several cell types, and known to play a role in embryogenesis, wound healing, angiogenesis, and tumorigenesis. The aim of this work was the functional and molecular characterization of TSP interactions with mesangial cells. Adhesion of mesangial cells to TSP-coated plastic, and chemotaxis in the Boyden chamber assay were tested. In order to identify TSP active domains, heparin, known to bind to the amino-terminal region of TSP, four monoclonal antibodies directed against specific domains of the molecule, and TSP fragments obtained by enzymatic digestion were used. We found that TSP induces mesangial cell adhesion and chemotaxis, in a dose dependent manner. Adhesion was inhibited by antiserum against TSP, and by an anti-CD36 monoclonal antibody tested in the presence of heparin, but not by the peptide Gly-Arg-Gly-Asp-Ser. We have also found that only the carboxy-terminal end of TSP retains the adhesive properties of the molecule, while all the fragments tested showed some degree of chemotactic activity. We conclude that TSP modulates mesangial cell adhesion and motility, thus acting as a potential autocrine and paracrine regulator of mesangial cell functions in normal and pathologic conditions.


Pathophysiologic implications of proteinuria in a rat model of progressive glomerular injury. Males of a substrain of Munich-Wistar rats (MWF/Ztm), selected for their large number of superficial glomeruli, develop spontaneous proteinuria with age, whereas females of this strain have a normal urinary protein excretion rate. We investigated the relationship between functional and structural glomerular alterations in four groups of male and female MWF/Ztm rats, respectively at 20 and 35 weeks of age. Systolic blood pressure, urinary protein excretion and composition of urinary proteins were periodically measured during the study. At the end of the observation period, renal function was evaluated with solute clearance technique and kidney tissue processed for morphologic and morphometrical analysis using light and electron microscopy. Systolic blood pressure in males was significantly higher than in females, and progressed toward systemic hypertension with age. Urinary protein excretion became spontaneously abnormal in males. About 50% of urinary proteins consisted of albumin, whereas 35% was a sex-dependent low molecular weight protein. Albumin excretion increased with age in these animals, whereas excretion of the sex-dependent protein decreased. Urinary protein excretion in females was significantly lower than in males of the same age, remaining near normal levels. No decline in renal function with age was observed in all animal groups. Glomerular structural alterations developed progressively with age in males, leading to important glomerular and tubular alterations. Female rats maintained almost intact glomerular morphology until the end of the study. Morphometric analysis showed an important increase in glomerular volume with age in males but not in females. This glomerular tuft enlargement was the result of an increase in the number of glomerular cells and a concomitant increase in cell volume. Male MWF/Ztm rats develop spontaneously systemic hypertension, proteinuria of glomerular origin, and glomerulosclerosis. Female rats develop less severe hypertension and are protected from proteinuria and glomerulosclerosis.


A molecular biologic study of extracellular matrix components during the development of glomerulosclerosis in murine chronic graft-versus-host disease. We studied the development of glomerulosclerosis in murine chronic graft-versus-host disease, a model for human systemic lupus erythematosus. The disease was induced in (C57BL10 x DBA/2)F1 hybrids by injection of DBA/2 lymphocytes leading to deposition of auto-antibodies in the glomeruli, and a lupus type of nephritis morphologically. We have determined the levels of mRNA coding for laminin (B1 and B2), a 67 kilodalton laminin binding protein, and types I and IV collagen, in control and graft-versus host disease mice at various times after disease induction. Laminin and collagen mRNAs were increased in whole kidneys 4 weeks after induction of the disease. At week 10, all animals displayed dramatic stimulation of alpha 1(I), alpha 1(IV), laminin B1, and B2 mRNAs. The 67 kilodalton laminin binding protein mRNA was also doubled from week 4 to 16. In isolated glomeruli, the mRNA level coding for laminin B2 was already significantly increased from week 8. This enhancement of laminin synthesis corresponds to the mesangial expansion and to the development of laminin-containing spike formations of the glomerular basement membrane at week 8. The expansion of the mesangial matrix in murine chronic graft-versus-host disease is caused at least in part, by an increased production of extracellular matrix components by glomerular cells. These results demonstrate that the increase of specific extracellular matrix components mRNAs precedes light microscopic changes. Quantitative evaluation of the mRNA levels coding for extracellular matrix proteins may reveal a useful method for the early detection of the development of glomerular sclerosis at the stage preceding the onset of anatomo-clinical changes.


A comparative analysis of glycosaminoglycans from human umbilical arteries in normal subjects and in pathological conditions affecting pregnancy. Vascular cells express different phenotypes in adult and fetal vessels, and the extracellular matrix they synthesize should reflect these differences. Alterations of vascular proteoglycan/glycosaminoglycan is verified in disorders such as hypertension and diabetes, and when occurring during pregnancy, they bring about structural changes to fetal vessels that often lead to impaired fetus growth. Yet there is little data about the extracellular matrix of an important human fetal vessel, the umbilical artery. This study involved the biochemical characterization of the extracellular matrix of normal umbilical arteries, umbilical arteries from complicated pregnancies (maternal hypertension and diabetes and intrauterine growth retardation syndrome), and, for purpose of comparison, normal adult arteries (aorta and iliac and pulmonary arteries). Although the collagen types I:III ratio was determined in some cases, emphasis was placed on analysis of glycosaminoglycans. Normal umbilical arteries differ from normal adult arteries in that they contain greater concentrations of hyaluronic acid and lesser concentrations of heparan sulfate and chondroitin 4- and 6-sulfate. The umbilical artery also differs from adult arteries in the disaccharide composition of its chondroitin and heparan sulfates and in the molecular weight of this latter glycosaminoglycan. The glycosaminoglycan distribution in umbilical arteries derived from complicated pregnancies is roughly similar to that of controls. However, total glycosaminoglycan and collagen were significantly reduced, and the collagen I:III ratio was increased in the umbilical arteries from hypertension-complicated pregnancies. The glycosaminoglycan composition of the normal umbilical artery, a fully differentiated tissue, differs in many aspects from that of normal adult arteries. Of the cases of complicated pregnancies studied, the extracellular matrix of umbilical arteries was altered only in maternal hypertension. The changes, notably a mild fibrosis, were not very pronounced and should not impair hemodynamic properties of the vessel.


Thrombin enhancement of interleukin-1 and tumor necrosis factor-alpha induced polymorphonuclear leukocyte migration. Cytokines such as IL-1 alpha and tumor necrosis factor-alpha (TNF-alpha) activate vascular endothelium to express leukocyte adhesion molecules that promote polymorphonuclear leukocyte (PMNL) migration and to synthesize tissue factor, thus making the endothelium a procoagulant surface. alpha-Thrombin, generated during coagulation, also activates endothelial cells. Since all these processes are likely involved in inflammation, the effect of alpha-thrombin on PMNL interaction with cytokine activated endothelium was investigated. Human umbilical vein endothelium was grown on polycarbonate filters to investigate the effects interleukin-1 alpha (IL-1 alpha), TNF-alpha, and alpha-thrombin on PMNL transendothelial migration quantitated with 51Cr-labeled PMNL, and on endothelial monolayer permeability, quantitated with 125I-labeled albumin (HSA). To evaluate the expression of endothelial-leukocyte adhesion molecules, enzyme-linked immunosorbent assay was performed on human umbilical vein endothelium monolayers. The effect of thrombin on PMNL accumulation and plasma exudation in inflammation was studied in a rabbit dermal model, using 51Cr-labeled blood leukocytes and [125I]HSA respectively. On resting human umbilical vein endothelium, alpha-thrombin induced a transient increase (2.5- to 4-fold) in monolayer permeability lasting 30 minutes. Slight but significant transendothelial migration of 51Cr-labeled PMNL was induced by alpha-thrombin (7.4 +/- 0.6% of cells added, unstimulated = 1.9 +/- 0.4%), although this response was less than that induced by f-norLeu-Leu-Phe (17%), IL-1 alpha (29%) or TNF-alpha (21%). alpha-Thrombin enhanced the initial rate of IL-1, TNF-alpha and f-norLeu-Leu-Phe induced PMNL transendothelial migration in an additive or supradditive manner (e.g., with IL-1 alpha+alpha-thrombin, migration was 58% greater than additive at 15 to 30 minutes, p < 0.001). Catalytically inactivated alpha-thrombin, D-phenylalanyl-L-propyl-L-arginine chloromethyl ketone and diisopropyl-fluorophosphate alpha-thrombin, did not enhance migration or permeability. In dermal inflammation in rabbits, alpha-thrombin (10 units/site) induced an increase in plasma protein exudation, with only a mild infiltration of PMNL. However, alpha-thrombin synergistically enhanced the PMNL infiltration induced by IL-1 alpha, TNF-alpha, but not that induced by zymosan activated plasma (C5a) or IL-8 (neutrophil-activating peptide-1). These measurements were confirmed histologically. Investigations into the mechanisms of the enhancement of PMNL migration indicated that individually vascular permeability changes, prostaglandins, platelet activating factor, and P-selectin expression did not account for the observation effects. Alpha-thrombin may have a role in synergistically enhancing PMNL infiltration at sites of inflammation, in part via enzymatic action on the cytokine activated endothelium. The mechanisms involved in this effect are likely a complex interaction.


Langerhans cells, macrophages and lymphocyte subsets in the cervix and vagina of rhesus macaques. The lower reproductive tract is an important site of contact with pathogenic microorganisms and local immune responses to a variety of antigens have been reported. The purpose of this investigation was to define the morphology of the mucosa-associated lymphoid tissue in the genital tract of rhesus monkeys. Monoclonal antibodies were used in an immunoperoxidase staining technique to identify immunophenotypic markers on mononuclear cells in the vaginal and cervical mucosa of 14 cycling, multiparous rhesus macaques. CD1a+ Langerhans cells were present in the stratified squamous epithelium of the vagina (14 animals) and ectocervix (11 animals). Surprisingly, CD1a+ dendritic cells were also found within the columnar epithelium of the endocervix (5 animals). Moderate numbers of CD68+ macrophages were located in the submucosa of the vagina, ectocervix, and endocervix of all the monkeys. In all of the animals, moderate numbers of CD8+ lymphocytes were present in the submucosa and squamous epithelia of the vagina and ectocervix. Variable numbers of CD20+ B cells and CD4+ lymphocytes were located in the submucosa of all the areas examined. Lymphoid nodules were present in the submucosa of vagina (14 animals) and ectocervix (4 animals), and these nodules contained macrophages, CD4+ T cells and B cells, with fewer numbers of CD8+ T cells and Langerhans cells. These observations provide a morphologic basis for mucosa-associated lymphoid tissue in the female genital tract. Langerhans cells in the vaginal mucosa and endocervix may be well suited to sample antigen in the lumen of the reproductive tract, travel to the draining lymph node, present the antigen to T lymphocytes and initiate an immune response. This pathway of antigen presenting cell migration from the mucosa to the genital lymph node may represent the inductive arm of the mucosal immune system in the lower female reproductive tract.


Astrocyte activation correlates with cytokine production in central nervous system of Trypanosoma brucei brucei-infected mice. During the late-stage disease associated with human African trypanosomiasis, caused by infection with either Trypanosoma gambiense or T. rhodesiense, parasites invade the central nervous system (CNS), eventually leading to development of CNS pathology. This can be exacerbated by subcurative chemotherapy. The mechanisms through which the inflammatory processes within the CNS are controlled remain unclear. Mice infected with T. b. brucei were treated with a trypanocidal drug regimen on day 28 postinfection that cleared parasites from all sites except the brain. Brains of mice killed at different times during infection and after chemotherapy were analyzed, using immunocytochemistry for astrocyte activation and polymerase chain reaction assisted amplification of RNA to detect cytokine transcripts. Drug-treated animals developed a posttreatment meningoencephalitis similar to that which can occur in humans with late-stage African trypanosomiasis. Between days 14 and 21 postinfection, before chemotherapy and the subsequent development of inflammatory lesions in the brain, astrocytes became activated. The production of several cytokines correlated with this astrocyte activation. Low levels of interleukin-1 alpha transcripts were detected in uninfected controls, but levels increased with astrocyte activation in the infected animals. Transcripts for the macrophage inflammatory protein-1 and tumor necrosis factor-alpha were first detected on day 21 postinfection, with higher levels in mice after development of the posttreatment meningoencephalitis, whereas granulocyte macrophage-colony stimulating factor was detected only in animals that developed posttreatment reaction. Interleukin-6 and interferon-gamma were also first detected on day 21 postinfection, correlating with astrocyte activation but subsequently declined with time in both treated and untreated mice. These results indicate that cytokines are being produced within the CNS before any inflammation and that astrocytes may be the source of some of these cytokines. Thus astrocyte activation may be key in the control and development of the CNS inflammatory processes that occur in African sleeping sickness.


Characterization of high affinity binding between laminin and Alzheimer's disease amyloid precursor proteins. In vivo amyloid formation apparently involves several extracellular matrix components that are usually found associated with basement membranes. These include laminin, heparan sulfate proteoglycan, collagen type IV, and entactin. These components have also been found in neuritic plaques. We have therefore been examining interactions between extracellular matrix components and the Alzheimer's amyloid precursors (AAPs). Binding interactions of laminin with AAP-695, -751, and -770 were examined using a solid phase enzyme-linked immunosorbent assay technique. Objective, quantitative analyses of the laminin AAP-695, -751, and -770 binding data reveal two binding sites for laminin, with Kd values of 1 x 10(-10) M and 1 x 10(-8) M. Zinc and dithiothreitol profoundly stimulate laminin binding to AAPs. Furthermore, zinc fingers were found in the laminin amino acid sequences. Previous binding studies of AAPs with the basement membrane heparan sulfate proteoglycan revealed similar affinities. A particular order of addition of laminin and heparan sulfate proteoglycan to AAPs can be demonstrated. These avid interactions with extracellular matrix proteins likely reflect normal functions of the AAPs and may be involved in nucleation events in Alzheimer-type amyloid formation.


A new marker for rhabdomyosarcoma. Insulin-like growth factor II. Accurate diagnosis of soft tissue tumors is often difficult and considered important because the clinical outcome and the modality of therapy vary according to tumor type. We have realized that rhabdomyoblasts seen in occasional Wilms' tumors express high levels of insulin-like growth factor II (IGF-II) transcripts. This observation prompted us to see if IGF-II could be a useful marker for differential diagnosis of rhabdomyosarcoma (RMS) from other soft tissue sarcomas. Five cases of RMSs (including embryonal, botryoid, and alveolar types), various other soft tissue tumors, and 10-week gestation fetal muscle tissue were studied to demonstrate IGF-II transcripts by means of Northern blot hybridization and in situ mRNA hybridization using radio-labeled probes. Hybridization results were then compared with data obtained from immunohistochemistry using antibodies against desmin and vimentin. Northern blot hybridization demonstrated that all five RMSs contained three major sizes of IGF-II transcripts. In situ hybridization indicated that all RMSs showed highly strong and specific IGF-II mRNA expression, whereas other soft tissue tumors showed very low or no signal. A characteristic finding was that the expression of IGF-II mRNA was inversely correlated with the degree of tumor cell differentiation. Thus poorly differentiated RMSs showed the highest signal, whereas well-differentiated RMSs showed low expression, albeit, still significantly higher than normal skeletal muscle fibers. Among five RMSs, desmin was seen in three tumors and vimentin in all tumors. Desmin was negative in other nonskeletal muscle origin tumors while vimentin was present in all tumors examined. IGF-II has potential as a new marker for differential diagnosis of RMS from other soft tissue tumors and could be especially useful in differential diagnosis of small blue cell tumors.


Multiple-modified desialylated low density lipoproteins that cause intracellular lipid accumulation. Isolation, fractionation and characterization. The basic differences between sialylated (sialic acid rich) and desialylated (sialic acid poor) human low density lipoproteins (LDL) are not fully defined. It is not known whether there are any differences in the LDL composition of coronary atherosclerosis patients and healthy individuals. Sialylated (45 to 94% of total LDL) and desialylated (6 to 55%) LDL were separated by affinity chromatography on Ricinus communis agglutinin-agarose, and their chemical composition and physical properties were examined. Sialic acid contents in sialylated LDL fractions of healthy subjects and patients were the same and 1.5 to 3-fold higher than in desialylated LDL. Desialylated LDL had smaller sizes and greater electrophoretic mobility than sialylated ones. Desialylated, but not sialylated LDL, induced 1.5- to 4-fold accumulation of neutral lipids in human aortic smooth muscle cells and human blood monocytes. Subfractions of desialylated LDL containing lower amount of sialic acid revealed higher ability to accumulate lipids in cultured cells. Desialylated LDL contained lower amounts of cholesteryl esters, free cholesterol and triglycerides as compared with sialylated LDL. On the other hand, concentration of di-, monoglycerides and free fatty acids in desialylated LDL was 2 to 3-fold higher than in sialylated lipoproteins. Desialylated LDL fraction was characterized by lower levels of phosphatidylcholine, sphingomyelin, phosphatidylethanolamine, but higher content of lysophosphatidylcholine. Freshly isolated sialylated and desialylated LDL contained equal amounts of thiobarbituric acid reactive substances, but oxidation of desialylated LDL was more pronounced in presence of Cu(2+)-ions. Desialylated LDL had higher level of oxysterols and lower amounts of vitamin A and E. Content of free amino groups of lysine in desialylated LDL of patients was 2-fold lower than in sialylated LDL. This difference was partially due to masking of amino groups caused by conformational change in the tertiary structure of apolipoprotein, partially to chemical modification of amino groups. When subfractionated by density gradient ultracentrifugation, desialylated LDL was represented by higher density particles than sialylated LDL. Sialic acid content in desialylated LDL subfractions decreased with rise of lipoprotein density. Higher density desialylated LDL and in less extent sialylated LDL contained smaller amounts of free and esterified cholesterol and phospholipids. Only the densest subfractions of desialylated LDL from healthy subjects caused intracellular lipid accumulation. Ability of patients' desialylated LDL to accumulate cholesterol in cells increased with particle density. Extensive biochemical and biophysical analysis performed in this study shows that desialylated LDL differ from these sialylated LDL in many respects. The LDL of coronary atherosclerosis patients differ from those in healthy individuals in several parameters.


Use of a school referendum to deter teen-age tobacco use. Montana's 1991 school "tobacco referendum," authorized by the state legislature and approved by about 60% of approximately 50,000 students voting in grades 7-12, represents an innovative attempt to include youth in decisions regarding tobacco and health and to decisively demonstrate peer disapproval of tobacco use. The referendum asked students whether tobacco sellers should voluntarily refuse to sell cigarettes and tobacco to persons younger than age 18. Montana is one of five states which allow tobacco sales to minors but has one of the nation's lowest youth smoking rates. The statewide referendum follows the successful example of the Bozeman, Montana, school district's 1990 tobacco referendum, in which 80% of the 2,200 grade 7-12 students and staff voted to make their schools "tobacco-free."


Susceptibility to peer pressure as an explanatory variable for the differential effectiveness of an alcohol misuse prevention program in elementary schools. A school-based alcohol misuse prevention program had differential effects on students' susceptibility to peer pressure, depending on prior experience with alcohol. These effects paralleled those on alcohol use and misuse, indicating program effects on use and misuse were mediated by reductions in the rate of increase on susceptibility to peer pressure. Experimental group students with prior unsupervised use of alcohol showed a significantly greater reduction than their controls in the rate of increase in susceptibility to peer pressure, alcohol use, and alcohol misuse. This difference was not found among students without prior unsupervised use of alcohol.


Peer perceptions of adolescent health behaviors. Perceptions adolescents form of peers in relation to modeled health behaviors were examined. Five hundred ten adolescents, ages 12-15, from eight midwest schools were shown a slide of a male or female adolescent displaying a health behavior artifact (apple, tennis racket, cigarette, beer can), or without an artifact and asked to rate the model on 16 characteristics using a semantic differential scale. Data were factor analyzed using principle components analysis and a 2x2x5 MANOVA. Results indicated that models appeared less mature when holding a beer can or cigarette. In addition, the female model was rated more popular than the male in the control and beer can depictions. The influence of modeled behaviors on traits adolescents want to develop must be understood to present effective health education programs. Educational efforts should include consideration of the perceptions adolescents hold of their peers' health behaviors.


Demographics of adolescent sexual behavior, contraception, pregnancy, and STDs. The demographics of fertility-related behavior of youth ages 10-18 are reviewed. Data were collected from U.S. vital statistics, and birth rates, contraceptive use, sexual behavior, number and types of sexual partners, patterns of sexual initiation and sexual intercourse, and sexually transmitted diseases were examined. Despite data limitations, the demographic profile of adolescent sexual intercourse is striking. Age clearly is the single most important predictor of sexual debut.


Risk factors for adolescent sexual behavior, fertility, and sexually transmitted diseases. Current understanding of the risk factors related to adolescent initiation of sexual activity, use of contraception, pregnancy, and STDs is examined. From recent research on adolescent fertility, findings that have particular relevance to school health or reflect new understandings of adolescent sexuality are summarized. In selected cases, prevention programs that build directly on an understanding of these risk factors are cited.


School-based programs to reduce sexual risk-taking behaviors. This article reviews the major approaches implemented during the last two decades to reduce sexual risk-taking behaviors, examines their evidence for success, and provides several recommendations for effective programs and program evaluations. This article does not discuss more broad-based sexuality education programs which address sexuality in a broader context. Instead, this article focuses primarily on programs that educators believed would reduce unprotected sexual intercourse.


Teen contraception in the 1990s.

Teen contraception in the 1990s. Basic information about appropriate contraceptive methods for teen-agers is provided. Methods most commonly used will be reviewed and an overview of alternative methods available for use in special subgroups will be provided.


Adolescent reproductive health: roles for school personnel in prevention and early intervention. The practical roles school staff can play in addressing adolescent reproductive health issues such as teen-age pregnancy, STDs, and HIV/AIDS are reviewed. Particular attention is given to identification and assessment of adolescents most at risk for pregnancy, STDs, and HIV/AIDS, and the regulations that address protection of students' privacy in this area.


Adolescent pregnancy options.

Adolescent pregnancy options. The range of pregnancy options available to adolescents each have significant ramifications for future educational and economic achievement. The changing societal context of adolescent pregnancy decision-making are described, and the characteristics of adolescents who choose to terminate their pregnancy, parent their child, or place for adoption are examined. The role of significant others in decision-making and the implications of mandatory parental involvement in pregnancy decision-making is discussed, as well as the roles of schools in promoting the well-being and potential of adolescents considering pregnancy decisions.


School-based prenatal and postpartum care: strategies for meeting the medical and educational needs of pregnant and parenting students. Adolescent pregnancy is not a new phenomenon, but it is a source of concern to U.S. educators because of the many young women choosing to become parents and the profound effect early childbearing has on the educational and vocational careers of many young Americans. Strategies for addressing the unique medical, educational, and psychosocial needs of pregnant and parenting students will be examined. Documented physical and psychosocial risks associated with adolescent childbearing will be reviewed, and data related to the effect of young maternal age on pregnancy outcome and the effect of pregnancy on the life course development of adolescents will be emphasized. Specific elements of comprehensive, adolescent-oriented prenatal and postnatal care will be discussed, as well as the effectiveness of existing prenatal and postnatal programs at preventing the most serious sequelae of adolescent childbearing. The role of school-based services will be examined, and ways will be discussed for educators and health care providers to collaborate in providing medical, educational, and social services for adolescent parents and their children. In addition, topics for future research will be suggested.


Consequences of teen-age parenting.

Consequences of teen-age parenting. The changing context of teen-age childbearing and current related controversies are reviewed. Recent research about the consequences of teen-age childbearing for the teen-age mother, the father, and for the children born is examined. The article also summarizes current knowledge about the consequences of teen-age childbearing with regard to the mother's educational attainment, marital experience, subsequent fertility behavior, labor force experience and occupational attainment, and experience with poverty and welfare.


Abortion among adolescents: research findings and the current debate. Utilizing research that focuses on adolescents as well as findings in samples which might have special relevance to young, unmarried women, this report summarizes research on the consequences of abortion among adolescents. It discusses prior literature in the area of parental notification and parental consent, subjects on which public opinion is not divided along familiar pro-choice/anti-choice lines. Following a discussion of methodological problems identified in prior research, it reports on a study designed to address these problems in an adolescent population; it discusses implications for the current debate of this and other studies' findings that there are no identifiable adverse sequelae of the abortion process.


Epidemiology of AIDS, HIV prevalence, and HIV incidence among adolescents. Health educators, policy analysts, and public health officials are becoming more aware of the serious threat HIV poses to the health of U.S. adolescents. While AIDS among adolescents remains relatively uncommon, considerable data indicates this age group has alarmingly high HIV infection rates and that minority adolescents are at disproportionately greater risk of HIV infection relative to their White peers. Recent seroconversion studies of active duty military personnel indicate that the number of new HIV infections (incident cases) are especially high among Black adolescents. Findings suggest the urgent need for more tailored HIV prevention programs, especially gender- and ethnic-specific programs.