Adenoid cystic carcinoma of the head and neck: evaluation with MR imaging and clinical-pathologic correlation in 27 patients. Twenty-seven adenoid cystic carcinomas (ACCs) of the head and neck in 27 patients were evaluated in a retrospective study based on findings at magnetic resonance (MR) imaging and pathologic and clinical examination. Clinical follow-up was obtained over a mean period of 6.3 years (range of follow-up, 3 months to 17 years); all patients underwent one to seven MR examinations. On T2-weighted images, lesions with low signal intensity corresponded to highly cellular tumors (solid subtype) with a poor prognosis; lesions with high signal intensity corresponded to less cellular tumors (cribriform or tubular subtype) with a better prognosis. MR images were not specific in differentiation of ACCs from other types of tumors; this result underscores the need for biopsy to ensure correct diagnosis. Local, intracranial, osseous, and perineural invasion was depicted, but because of its lack of specificity, MR imaging caused overdiagnosis of tumor extension, particularly perineural spread and bone abnormalities.


Synthesis of prostaglandins by pig blastocysts cultured in medium containing estradiol or catechol estrogen. Two experiments were conducted to determine the effects of 2-hydroxy-estradiol-17 beta (2-OH-E2; 0, 50 and 100 microM) and estradiol-17 beta (E2; 0, 25 and 50 microM) on prostaglandin (PG) E and PGF2 alpha synthesis by day-10 pig blastocysts (day 0 is first day of estrus). Blastocysts were incubated in a modified Krebs-Ringer bicarbonate medium, supplemented with bovine serum albumin (4 mg/ml) and the vitamins and amino acids (essential and nonessential) in Minimum Essential Medium (without phenol red or antibiotics). The incubations were conducted at 39 degrees C for three 2-h periods; the second and third periods included an E2 or catechol estrogen treatment. Release of PGF2 alpha into the culture medium decreased (p less than 0.001) linearly with increasing concentrations of 2-OH-E2 in both periods. Release of PGE was not affected by 2-OH-E2, therefore 2-OH-E2 increased (p less than 0.06) the PGE:PGF2 alpha. When E2 was added to the medium, release of PGE was decreased (p less than 0.01) during the second and third periods. Release of PGF2 alpha also was decreased (p less than 0.05) by E2 during period 2, but E2 did not alter the PGE:PGF2 alpha. Content of PGs in blastocysts at recovery was less than 10% of the PGs released in vitro. Therefore, these studies demonstrate effects of both the primary and catechol forms of E2 on the synthesis of PGE and PGF2 alpha. Catechol estrogens and E2 may inhibit PG synthesis and modify the PGE:PGF2 alpha during the establishment of pregnancy in pigs.


Radiotherapy enhanced by cis-platinum in stage III non-small cell lung cancer: a phase II study. From January 1984 to December 1986, 94 patients with unresectable, locally advanced, non-small cell lung cancer (NSCLC) were treated to assess both the efficacy and the toxicity of a combined modality treatment including radiation therapy (45 Gy/15 fractions/3 weeks) and daily low dose cDDP (6 mg/m2). The overall response rate for the 90 evaluable patients was 54.3% with 16.6% of complete responses. At a minimum follow-up of 4 years, the overall median survival time was 12 months. Provided adequate hydration is ensured, the cDDP regimen chosen as a radiosensitizer can be safely combined with radiation therapy.


Differential influence of prognostic factors on the occurrence of metastases at various anatomical sites in human breast cancer. The present study was initiated by data on the anatomical occurrence of metastases from breast cancer. Metastases may occur at various organs, here grouped into ten categories, and simultaneous occurrences at several sites are common. Our aim was to identify and compare risk factors for development of metastases at each site. The influence of the various risk factors for recurrence at one specific site can be modelled as a standard competing risk problem, using well-established survival analysis techniques such as the Cox regression model. Recently a generalization to joint occurrences at more than one site has been proposed. The more general models allow comparison of the differential prognostic influence of various risk factors on recurrence at different sites. We applied these methods to data from the Danish Breast Cancer Cooperative Group on recurrence after breast cancer, and examined the effect of degree of anaplasia, number of positive lymph nodes, size of primary tumour, skin or deep fascial invasion, age of the patient and adjuvant treatment regimen, as possible risk factors. There were no differential effects of type of therapy on recurrence in different sites. However, chemotherapy had a marginally significant positive effect at all sites. Moreover the number of positive lymph nodes was associated with an increased risk of metastases, homogenous across sites, except for brain where an opposing trend was found. Similarly the degree of anaplasia was associated with a somewhat increased risk of metastases, homogenous except for brain. For brain metastases a significantly more pronounced effect of degree of anaplasia was found. Comparison of the results obtained from the two types of models reveals that the new more general models confirm fewer differences between the influence of the prognostic factors than the traditional regression models suggest.


Histological distribution of the interleukin-4 receptor (IL4R) within the normal and pathological synovium. The authors sought the synovial distribution of the interleukin-4 receptor in the synovium of 3 normal subjects, 5 cases of osteoarthritis (OA) and 10 cases of rheumatoid arthritis (RA) and various other inflammatory arthritic conditions. The specific receptor for interleukin-4 was mainly found on cells around subintimal blood vessels and within lymphocytic aggregates where dendritic-like cells were strongly stained. Conversely, synovial lining cells did not express the interleukin-4 receptor. In conclusion, it is believed that the expression of interleukin-4 receptor is restricted to the early activation stage of the mononuclear cells in the pathological joint.


Strand-specific recognition of a synthetic DNA replication fork by the SV40 large tumor antigen. The mechanism by which DNA helicases unwind DNA was tested; an "unwinding complex" between the SV40 large tumor antigen (T antigen) and a DNA molecule designed to resemble a replication fork was probed. In an adenosine triphosphate (ATP)-dependent reaction, T antigen quantitatively recognized this synthetic replication fork and bound the DNA primarily as a hexamer. The T antigen bound only one of the two strands at the fork, an asymmetric interaction consistent with the 3'----5' directionality of the DNA helicase activity of T antigen. Binding to chemically modified DNA substrates indicated that the DNA helicase recognized the DNA primarily through the sugar-phosphate backbone. Ethylation of six top strand phosphates at the junction of single-stranded and double-stranded DNA inhibited the DNA helicase activity of T antigen. Neither a 3' single-stranded end on the DNA substrate nor ATP hydrolysis was required for T antigen to bind the replication fork. These data suggest that T antigen can directly bind the replication fork through recognition of a fork-specific structure.


Treatment of relapse of small cell lung cancer in selected patients with the initial combination chemotherapy carboplatin, etoposide, and epirubicin. Relapse of small cell lung cancer is not usually treated with further chemotherapy as results are considered to be disappointing. Six patients with relapse of small cell lung cancer after a complete initial response and remission of more than one year responded to repeat treatment with the initial chemotherapy comprising carboplatin, etoposide, and epirubicin. The second remission ranged from six months to more than 15 months. In patients with a relapse of small cell lung cancer after a complete initial response and prolonged remission retreatment with the initial combination chemotherapy cannot be dismissed and requires further study.


[Effect of feeding on the energy balance and fertility in dairy cows]. The effect of two different feeding regimes on energy metabolism and fertility was studied in dairy cows. The cows were divided according to the type of feed into a hay-urea and a silage group. Ketone body levels in blood were used as indicators of energy balance. Intake of dry matter and of energy was lower in the silage group than in the hay-urea group. The energy balance had almost equilibrated in the hay-urea group at 50 days p.p., in contrast to that of the silage group. The ketone body levels were lower in the hay-urea group than in the silage group in early lactation. The cows in the hay-urea group had a shorter interval from calving to first insemination and to conception than those in the silage group. Low energy level in the late puerperal period had an adverse effect on fertility. The higher the level of ketone bodies at day 50 p.p., the lower the fertility of the cows is likely to be.


[The effect of different zootechnical practice methods on the duration of labor in swine with reference to the body condition of the sow]. In a single large pig production unit of 6000 breeding sows, seven groups of 100 sows each were formed at random on the 110th day of pregnancy. Each group was evaluated and divided according to body condition in three subgroups. The groups were treated as follows: Group 1 received a single 3 mg intramuscular dose of alfaprostol on the 113th day of pregnancy; Group 2 received a single 3 mg intramuscular dose of alfaprostol on the 113th day of pregnancy, followed 24 hours later by a single intramuscular dose of 10 IU oxytocin; Group 3 received a single intramuscular dose of 10 IU of oxytocin after the birth of the first piglet; Group 4 received a single 100 mg intramuscular dose of prednisolone on 113th day of pregnancy; Group 5 received a single 0.2 mg intramuscular dose of carbamylcholine on the 113th day of pregnancy; Group 6 received feed containing 12% fiber, at 3 kg per sow per day from the 110th day of pregnancy; Group 7 received a 3 ml dose of physiologic NaCl solution i.m. on the 114th day of pregnancy. The parameter "duration of parturition" was evaluated. Groups 4 (prednisolone), 5 (carbamylcholine) and 6 (fiber) showed a shorter duration of parturition when compared to groups 1, 2, 3 and 7. The subgroups 5 in each group showed a longer parturition time, when compared to subgroups 3 and 4.


[Trichomonad infections of the oral cavity in cats in south Germany]. In this investigation trichomonads were isolated from the oral cavity in 21 of 110 examined cats, and only from those which were simultaneously FeLV, FIV or FIP positive. By means of scanning electron microscopy the trichomonads were shown to be round or piriform parasites which were on average 7.9 microns long and 6.2 microns wide at maximum width. They had 4 anterior flagella, which were on average 9.4 microns long, an undulating membrane measuring 6.7 microns with no trailing flagellum as well as an axostyle extending on average 5.0 microns beyond the body, and therefore should be attributed to the genus Trichomonas.


Lead nephrotoxicity and associated disorders: biochemical mechanisms. Lead may exert toxic effects on several organ systems, but those in the kidney are the most insidious. Acute lead nephropathy is characterized by proximal tubular dysfunction with the development of a Fanconi-type syndrome, alterations in mitochondrial structure and the development of cytosolic and nuclear inclusion bodies. Intracellular lead is associated with specific high affinity proteins and can also bind to metallothionein. Chronic lead nephropathy is irreversible and is typically accompanied by interstitial fibrosis, both hyperplasia and atrophy of the tubules, glomerulonephritis and, ultimately, renal failure. In addition, lead produces renal neoplasms in experimental animals. Chronic lead exposure is also implicated in the development of saturnine gout and hypertension. The metal interacts with renal membranes and enzymes and disrupts energy production, calcium metabolism, glucose homeostasis, ion transport processes and the renin-angiotensin system. This review summarizes the biochemical effects of lead on the kidney to understand the mechanisms of lead-induced nephropathy and other associated disorders.


Malignant myositis ossificans. A case report. A patient presented with an ossifying thigh mass suggestive of myositis ossificans. He had no antecedent trauma to the area. The mass was found to be an ossifying soft tissue metastasis from an occult gastric adenocarcinoma primary. Malignancy, and especially metastatic malignancy, is rarely considered in the differential diagnosis of a radiographic presentation of myositis ossificans.


Rhabdomyosarcomas developing in association with mediastinal germ cell tumours. Two mediastinal rhabdomyosarcomas that arose in association with germ cell tumours are reported. One presented as a small component of a mixed germ cell tumour with yolk sac and immature teratomatous elements. The other appeared as a large mass 4 months after diagnosis of a yolk sac carcinoma that had been treated with chemotherapy. The first patient was alive and free of disease 7 years later, whereas the second died of tumour 3 months post-operatively. The proportion of rhabdomyosarcoma within the germ cell tumours appears to have influenced the prognosis of these patients. This observation emphasizes the necessity of performing a thorough search for sarcomatous elements and quantifying their relative proportion in germ cell tumours of the mediastinum.


Intravenous anti-D gammaglobulin for the prevention of rhesus isoimmunization caused by platelet transfusions in patients with malignant diseases. Previous studies of sensitization to RhD by RhD-positive platelet transfusions in RhD-negative cancer patients have shown different frequencies of alloimmunization (max. 19%). We studied 37 RhD-negative patients who received RhD-incompatible platelet transfusions and simultaneously anti-D-immune globulin. We provide evidence that in this setting RhD-prophylaxis is highly effective in preventing alloimmunization due to RhD antigen, since none of the patients studied developed anti-D. Detection of other red blood cell antibodies than anti-D proves the possibility of immunization in these patients. Prevention of isoimmunization in patients with malignant diseases is recommended especially in young females, since an increasing number of patients are having successful pregnancies, despite prior or even during cytotoxic therapy.


Squamous cell carcinoma of the conjunctiva. Failure to demonstrate HPV DNA by in situ hybridization and polymerase chain reaction. Squamous cell carcinoma of the conjunctiva is a distinct rarity, often arising at the corneoscleral limbus and initially resembling pterygium or chronic keratoconjunctivitis. In this paper we report 4 patients with conjunctival squamous cell carcinoma/carcinoma in situ, which comprise all the cases found in the files of Kuopio University Hospital during 1959-1991. The clinical appearance, diagnosis and treatment of the lesions are described. All biopsies were studied for the presence of Human papillomavirus (HPV) DNA (recently demonstrated in conjunctival squamous cell papillomas, precancer lesions and carcinomas) by using in situ DNA hybridization (ISH) and polymerase chain reaction (PCR). Both techniques failed to demonstrate the DNA of any of the following HPV types: HPV 6, 11, 16 and 18 in any of the lesions. The results are discussed in the light of the recently proposed HPV etiology of these lesions.


Establishment and characterization of cell lines from human adenovirus type 12-induced murine tumors producing endogenous virus particles. Two cell lines designated IC-KMS and D-KMS were established from human adenovirus type 12-induced tumors of C3Hf/OK mouse. The cell lines retained the characteristics of the original tumor i.e., production of numerous C-type and intracisternal A-type particles, integration of Ad12 E1 region DNA and amplification of the myc gene family. Chromosomal analysis revealed chromosome aberrations in both IC-KMS and D-KMS cells. The modal chromosome number of IC-KMS cells was 54 and that of D-KMS cells was 48. Metacentric chromosomes and minichromosomes were found. Trisomy of chromosome 3, 7 and 12 was seen frequently in D-KMS cells. Although DNA aneuploidy was revealed by flow cytometry, the DNA indices of these cells showed no relation to the copy number of integrated Ad12 DNA. These cells have been propagated by serial culture during the past 17 months. Production of endogenous virus particles is a unique characteristic of IC-KMS and D-KMS cells. These cell lines would be useful materials for examining the contribution of Ad12 carcinogenesis to activation of endogenous virus particles, and also the correlation between Ad12 carcinogenesis and cancer-related genes.


Disseminated Pneumocystis carinii infection in a hemophiliac patient with acquired immunodeficiency syndrome. A case of disseminated Pneumocystis carinii (PC) infection in a 28-year-old Japanese male hemophiliac with acquired immunodeficiency syndrome (AIDS) is reported. The patient had displayed a high fever and diffuse faint interstitial infiltrates on chest X-ray films without dyspnea three months before his death. At that time, no PC was detected after four consecutive induced sputum tests. Serum anti-cytomegalovirus (CMV) IgM was positive by EIA. No treatment for PC and CMV was given at the patient's request. Autopsy findings disclosed disseminated PC infection consisting of granulomas with caseation-like necrosis and frothy exudate in the lungs and disseminated organized calcification in the blood vessels of extrapulmonary organs. PC cysts and/or trophozoites were detected in these lesions.


[Intracranial hemodynamics in phenobarbital administration]. The effect of phenobarbital application on cerebral hemodynamics was studied in premature lambs. Near infrared spectroscopy was used for noninvasive monitoring of oxyhemoglobin, deoxyhemoglobin, total hemoglobin and cytochrom a/a3. Total intracranial blood volume decreased shortly after giving the drug and remained then on a stable lowered niveau. This could be explained with shifting of blood in different brain regions or with a decreased total intracranial blood volume.


Jejunal perforation associated with cytomegalovirus infection in a patient with adult T-cell leukemia-lymphoma. A patient with adult T-cell leukemia-lymphoma suffered a jejunal perforation, which we believe was directly attributable to cytomegalovirus (CMV) infection. In the areas of ulceration and perforation in the small bowel, blood vessels penetrating the muscularis propria showed extensive lining of cytomegalic endothelial cells with CMV inclusions, accompanied by occasional disruption of the walls, partial occlusion of the lumina, fibrin thrombi, and hemorrhage. The CMV-induced vascular damage seemed to be closely related to the occurrence of ulcers and perforation. The recognition of CMV as a cause of lethal gastrointestinal lesions in immunocompromised hosts has become more important with the advent of anti-CMV therapy.


[Hemodynamic effects of stimulation of alpha 1-adrenoreceptors in healthy elderly and aged persons]. The effect of intramuscular administration of alpha 1-adreno-stimulator phenylephrine (standard dose of 0.15 mg/kg body wt.) on the chronotropic, pump, systolic, and diastolic heart function, renal blood circulation and microcirculation in different vessel regions was studied in 60 healthy human subjects of different ages (20-34, 60-74, and 75-89 years). In aging, the overall effect of standard phenylephrine dosage has been found to decrease. Parallel to this, there was a change in the structure of the response of cardiovascular system to alpha 1-adreno-stimulaor administration. In older population groups, the positive effect of phenylephrine upon the pump and systolic heart function decreased, while its negative effect on the renal blood circulation and microcirculation in different vascular regions increased.


Activation of muscarinic cholinergic receptors stimulates inositol phosphates synthesis in the developing avian cochlear duct. We previously reported that the inositol phosphates (IPs) synthesis is induced by muscarinic agonists in the rat cochlea and that this stimulation is maximal at postnatal day 12. This peak response is concomitant with the onset of the efferent synaptogenesis at the outer hair cell level. Whether the correlation between this neuronal plasticity and the enhanced IPs formation is unique to the rat or a general feature of the developing vertebrate cochlea is not known. To examine this question, we measured, in the presence of LiCl, the accumulation of (3H)-IPs induced by carbachol, in the developing chick cochlear duct during a period ranging from embryonic day (E) 8 to post-hatching day (P) 20. Carbachol (1 mM) causes a significant increase of IPs formation relative to basal values at all ages. This IPs accumulation is maximal at E8 (1854% of the basal level), then, rapidly decreases until P13 when it reaches a steady-state level of 294% of the basal level. Strikingly, this gradual decline in IPs formation is interrupted between E15 and E19, by a transient increase in IPs synthesis. This rise peaks at E16 with a stimulation value of 757% of the control level. This maximal stimulation is inhibited by atropine in a dose-dependent manner, as is the case at E9, suggesting the involvement of muscarinic receptors. Interestingly, the occurrence of the peak response is concomitant with the plastic events associated with the maturation of the efferent innervation of the cochlear duct. Thus, these results suggest that there may be a correlation between cochlear plasticity and enhanced IPs synthesis, which is not species-specific.(ABSTRACT TRUNCATED AT 250 WORDS)


Influence of prostaglandins on DNA and matrix synthesis in growth plate chondrocytes. Prostaglandins are locally produced in a number of tissues in response to a variety of stimuli, including local growth factors and systemic hormones. The present investigation characterizes prostaglandin effects on growth plate chondrocytes. Since cyclic adenosine monophosphate (cAMP) may act as a prostaglandin-stimulated second messenger, the effects of prostaglandins A1, D2, E1, E2, F2 alpha, and I2 (10(-10)-10(-6) M) on cAMP levels and thymidine incorporation were evaluated. The stimulation of cAMP and thymidine incorporation by the various prostaglandin metabolites were dose dependent and highly correlated (r = 0.99, p less than 0.001). The magnitude of the effect varied but was maximal at 10(-6) M for each of the prostaglandins. Prostaglandins of the E series (E1 and E2) were the most potent, causing significant effects at 10(-10) M and with maximal 12- and 13-fold increases in DNA synthesis after a 24 h exposure. Prostaglandins D2 and A1 maximally stimulated thymidine incorporation by 4.7- and 3.1-fold but caused significant increases only at 10(-8) M. Prostaglandins F2 alpha and I2 were the least stimulatory, producing small but significant increases in thymidine incorporation at 10(-6) M (30 and 100% stimulations). A causal relationship between cAMP and thymidine incorporation was further verified by the ability of dibutyryl-cAMP to increase DNA synthesis. Long-term chondrocyte cultures treated continuously with PGE2 demonstrated an increase in cell number, confirming the proliferative effect. Indomethacin did not alter the potent dose-dependent stimulations of chondrocyte DNA synthesis by TGF-beta 1, basic FGF, or PTH, indicating that these known mitogens act independently of prostaglandin metabolism. PGE2 was further examined for its effects of matrix synthesis. PGE2 inhibited collagen synthesis with a maximal 42% decrease but did not alter noncollagen protein synthesis. In contrast, PGE2 maximally increased sulfate incorporation by 35% and caused a small dose-dependent inhibition in alkaline phosphatase activity. Thus, prostaglandins alter DNA and matrix synthesis in growth plate chondrocytes and may have an important role in chondrocyte metabolism in the growth plate, fracture callus, and other areas of endochondral ossification.


Human cystatin C, a cysteine proteinase inhibitor, inhibits bone resorption in vitro stimulated by parathyroid hormone and parathyroid hormone-related peptide of malignancy. The effect of human recombinant cystatin C, a cysteine proteinase inhibitor, on bone resorption in vitro was evaluated. Bone resorption was assessed by analyzing the release of 45Ca and 3H from mouse calvarial bones prelabeled in vivo by injections with 45Ca or [3H]proline, respectively. In 24 h cultures, cystatin C (50 micrograms/ml) significantly inhibited the release of 45Ca and 3H stimulated by parathyroid hormone (PTH, 15 nmol/liter) or parathyroid hormone-related peptide of malignancy (PTHrP, 15 nmol/liter). The degree of inhibition caused by cystatin C in these 24 h cultures was similar to that caused by calcitonin (30 ng/ml). The inhibitory effect of cystatin C on 45Ca release induced by PTH was sustained in 96 h cultures, whereas the initial inhibition caused by calcitonin was transient. Cystatin C, 10-100 micrograms/ml, caused a dose-dependent inhibition of PTH (15 nmol/liter), and PTHrP (15 nmol/liter) stimulated 45Ca release. Addition of 50 micrograms/ml of cystatin C to mouse bone cultures inhibited the release of 45Ca induced by PTH and PTHrP at a wide range of submaximal and maximal concentrations of hormones (0.01-10 nmol/liter). No effect of cystatin C on 45Ca release in dead bones could be observed, nor did the inhibitor decrease the release of calcium in control bones. The inhibition by cystatin C on PTH-induced mineral mobilization was reversible. Cystatin C (1-100 micrograms/ml) did not affect protein synthesis or mitotic activities in mouse calvarial bones as assessed by the incorporation of [3H]proline and [3H]thymidine, respectively. These data show that cystatin C is a potent inhibitor of mineral mobilization and matrix degradation in cultured bones stimulated to resorb by PTH and PTHrP and that this effect is not due to general cytotoxicity.


1,25-Dihydroxyvitamin D3 analog structure-function assessment of the rapid stimulation of intestinal calcium absorption (transcaltachia). The possibility is now emerging that 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] can mediate biologic responses via both genomic and nongenomic pathways. To understand the molecular basis of the nongenomic response of transcaltachia, defined as the 1,25-(OH)2D3-mediated rapid (2-10 minutes) stimulation of calcium transport from the brush border to the basal lateral membrane of the epithelial cell in vitamin D-replete chick intestine, and to address the issue of whether the same receptor for the secosteroid serves as the signal transducer for both genomic and nongenomic pathways, we carried out structure-function studies using seven analogs of 1,25-(OH)2D3 with different affinities for the classic nuclear 1,25-(OH)2D3 receptor as measured by determination in a steroid competition assay of the relative competitive index (RCI). The RCI of 1,25-(OH)2D3 is by definition 100. 1,25-(OH)2D3 initiates transcaltachia within 2-10 minutes of vascular perfusion and yields a biphase response curve. Dose-dependent stimulations of Ca2+ transport by the seven analogs indicates that different structural features are essential for initiating the transcaltachic response as contrasted with binding to the classic nuclear receptor. Vascular perfusion with analogs AT (25-OH-16-ene-23-yne-D3) and Y (25-OH-23-yne-D3), which are known to activate Ca2+ channels but bind very poorly to the classic receptor (RCI less than 0.5), are efficient in stimulating Ca2+ transport. By comparison, compounds BT [1 alpha,24S-(OH)2-22-en-26,27-dehydrovitamin D3] and V (1,25-(OH)2-16-ene-23-yne-D3], which bind very well to the classic nuclear receptor (RCI 75-111) but do not activate Ca2+ channels, are inefficient in stimulating Ca2+ transport. These results indicate that the membrane components that respond to the analogs of 1,25-(OH)2D3 with activation of Ca2+ channels have a different ligand specificity than the classic nuclear receptor.


The bradykinin receptor of the rat uterus affects the ligand properties of charged mercurials. Using monovalent organic mercurials to modify the response to the oxytocic action of bradykinin (BK) on the rat uterus, a charge effect, related to the reagent applied, was noted. With neutral, and positively charged mercurials, a bimodal pattern of activity, causing stimulation of BK contraction at lower, and inhibition at higher concentrations was observed. With negatively charged mercurials transient stimulation occurred, inhibition only appearing after very large doses and long periods of incubation. Anionic charges of the bradykinin receptor affect the ligand properties of mercurials, repelling negative charges. By themselves, mercurials did not cause any response.


Olive oil, corn oil, and n-3 fatty acids differently affect lipids, lipoproteins, platelets, and superoxide formation in type II hypercholesterolemia. To evaluate which dietary fat may provide the best response in terms of plasma lipids and lipoproteins and also of platelet aggregability and superoxide formation by white blood cells, 12 type II patients were randomly allocated to three different diets, which provided polyunsaturated fatty acids (corn oil), monounsaturated fatty acids (olive oil), and a supplementation of ethyl esters of n-3 fatty acids to a prudent diet. Olive oil and, more significantly, n-3 ethyl esters lowered total cholesterol best (-2.2% and -5.8%, respectively); the latter diet, as expected, also significantly lowered triglyceridemia (-21.4%). The corn-oil diet exerted a small, statistically significant reduction of high-density-lipoprotein cholesterol (HDL) (-4.3%), and it also lowered plasma total apo B concentrations (-3.8%). n-3 ethyl esters significantly raised both total (+3.1%) and particularly HDL2 cholesterol (+24%). Platelet reactivity was insignificantly reduced by the three regimens, but all three significantly reduced thrombin-stimulated formation of thromboxane B2. Finally, only the n-3 fatty acid supplementation significantly reduced O2- generation by adherent monocytes. Dietary unsaturated fatty acids are generally effective on the plasma lipid and lipoproteins in type II patients, but significant differences may be found between the three tested regimens.


Cholesterol-lowering effects of psyllium-enriched cereal as an adjunct to a prudent diet in the treatment of mild to moderate hypercholesterolemia. Psyllium hydrophilic mucilloid, rich in soluble fiber, significantly lowers serum cholesterol concentrations. To investigate the lipid-lowering effects of a ready-to-eat cereal enriched with psyllium, we studied 44 hypercholesterolemic ambulatory individuals for 7 wk, using a randomized, double-blind, parallel design. After a 1-wk baseline period, subjects were randomly assigned to consume 114 g/d of a psyllium-flake or wheat-bran-flake cerearl for 6 wk adjunctive to an American Heart Association Step I diet. Serum high-density-lipoprotein cholesterol and triglycerides remained unchanged throughout the study; however, serum cholesterol and low-density lipoprotein (LDL) concentrations were significantly lower on psyllium than on wheat-bran cereal. Additionally, serum total and LDL cholesterol values were significantly lower than baseline values in the psyllium group. The dietary nutrient intake of the two groups did not differ except for soluble fiber. Adding psyllium-enriched cereal to a prudent low-fat diet may enhance dietary management of hypercholesterolemia.


Effect of oat bran on plasma cholesterol and bile acid excretion in nine subjects with ileostomies. A higher excretion of dry matter, fat, nitrogen, energy, and total bile acids in ileal effluents; a lower plasma low-density-lipoprotein (LDL) and total cholesterols (12.1% and 9.0% lower respectively); but no change in plasma high-density-lipoprotein (HDL) cholesterol or apolipoproteins A-I and B were observed in nine subjects with ileostomies when they consumed an oat-bran, bread-based, high-fiber diet (HFD) as compared with a wheat-flour, bread-based, low-fiber diet (LFD) for 3 wk with a crossover design. Of the nine subjects only the subjects with a low daily excretion of bile acids had an elevated excretion of total bile acids during the HFD compared with the LFD. Total cholesterol, LDL cholesterol, and apolipoprotein B in plasma also decreased by 11.3%, 15.3%, and 10.7%, respectively, after consumption of the HFD for 3 wk.


Coincidental finding of May-Hegglin anomaly in a patient with end-stage renal failure. We present two cases of May-Hegglin anomaly incidentally discovered in a patient and his brother during investigation of the patient for end-stage renal failure and workup for renal transplantation. Routine laboratory tests were performed and included a basically normal clotting profile. Ultrastructural studies of the May-Hegglin inclusions proved diagnostic, findings were compared with those of two similar granulocyte inclusion bodies, and nomenclature discrepancies that still exist in most references are again emphasized. The finding of the May-Hegglin anomaly in our patient appears to be incidental to the underlying renal disease. A successful renal transplant has been carried out in this patient. We now report on a patient and his brother in which the MHA was discovered during workup of the patient for end-stage renal failure and renal transplantation. No association between the underlying renal disease and the MHA could be demonstrated.


Exclusion of the gelsolin gene on 9q32-34 as the cause of familial lattice corneal dystrophy type I. Familial lattice corneal dystrophy type I (LCD1) is a localized form of inherited amyloidosis limited to the corneal stroma. Recently the Finnish form of hereditary amyloidosis with lattice corneal dystrophy has been shown to be due to a mutation in the gelsolin gene (G654----A; Asp187----Asn). In this paper we exclude the gelsolin gene as the cause of the autosomal dominant form of isolated LCD1.


Evidence, from combined segregation and linkage analysis, that a variant of the angiotensin I-converting enzyme (ACE) gene controls plasma ACE levels. The hypothesis of a genetic control of plasma angiotensin I-converting enzyme (ACE) level has been suggested both by segregation analysis and by the identification of an insertion/deletion (I/D) polymorphism of the ACE gene, a polymorphism contributing much to the variability of ACE level. To elucidate whether the I/D polymorphism was directly involved in the genetic regulation, plasma ACE activity and genotype for the I/D polymorphism were both measured in a sample of 98 healthy nuclear families. The pattern of familial correlations of ACE level was compatible with a zero correlation between spouses and equal parent-offspring and sib-sib correlations (.24 +/- .04). A segregation analysis indicated that this familial resemblance could be entirely explained by the transmission of a codominant major gene. The I/D polymorphism was associated with marked differences of ACE levels, although these differences were less pronounced than those observed in the segregation analysis. After adjustment for the polymorphism effects, the residual heritability (.280 +/- .096) was significant. Finally, a combined segregation and linkage analysis provided evidence that the major-gene effect was due to a variant of the ACE gene, in strong linkage disequilibrium with the I/D polymorphism. The marker allele I appeared always associated with the major-gene allele s characterized by lower ACE levels. The frequency of allele I was .431 +/- .025, and that of major allele s was .557 +/- .041. The major gene had codominant effects equal to 1.3 residual SDs and accounted for 44% of the total variability of ACE level, as compared with 28% for the I/D polymorphism.(ABSTRACT TRUNCATED AT 250 WORDS)


Linkage of a variant or attenuated form of adenomatous polyposis coli to the adenomatous polyposis coli (APC) locus. Adenomatous polyps are an intermediate in the pathway to colon carcinoma. An inherited disorder, familial adenomatous polyposis coli (APC), is characterized by hundreds to thousands of adenomatous polyps. A previously reported family had colon cancer associated with a low average but highly heterogenous number of colonic polyps, this phenotype mapped to the APC locus on 5q. Four new families have been ascertained in which the phenotypic pattern was different from classical polyposis but similar to that of the "prototype" kindred reported earlier. By multilocus linkage analysis, the gene responsible for the disease phenotype was mapped, with a high level of confidence, to the APC locus in two of the four families with the attenuated or variant form of polyposis (AAPC); the results for the two remaining kindreds were inconclusive. A combined maximum LOD score of approximately 7.6 at a recombination fraction of 0 was obtained when the results were summed over the four pedigrees with markers closest to the APC locus. The establishment of genetic linkage in such families may point to the APC locus as having a more significant role in inherited predispositions to colorectal cancer than was previously thought.


Variability in the clinical and pathological findings in the neuronal ceroid lipofuscinoses: review of data and observations. We reviewed the clinical and pathological data on 319 neuronal ceroid lipofuscinosis (NCL) cases to determine the degree of variability within the different forms and among and within families. Thirty-six cases (11.3%) were the infantile form; 116 cases (36.3%), late infantile; 163 cases (51.1%), juvenile; and four cases (1.3%), the adult form (Kufs disease). Clinical variability was found in all forms studied, but was most striking in the juvenile and late infantile forms of NCL. The expected initial findings of seizures, dementia, blindness, or motor impairment were evident in 255 cases (80%), and rarer, less typical initial neurological symptoms were seen mainly in the 64 cases (20%) of the juvenile form: behavior abnormalities (18/64), psychoses (12/64), neuropathy (2/64), involuntary movements (15/64), ataxia (9/64). Six juvenile and two adult cases had no detectable impairment of vision. All 319 NCL cases had skin or conjunctive biopsies or buffy coats that showed the characteristic ultrastructural abnormalities of NCL. Variability was evident in 16.7% in that a combination of fingerprint, curvilinear, and membranous profile inclusion bodies was observed in storage lysosomes, although one type of inclusion was distinctly predominant for each form. Postmortem examination of brains of 19 NCL cases (three with the infantile form, six with the late infantile form, nine with the juvenile form, and one with the adult form) revealed characteristic changes. Sixteen of the 19 NCL brains (84%) showed pathological variability in that they contained more than one kind of characteristic inclusion body in the neuronal lysosomal storage compartment. In all 19 NCL brains, small amounts of aging lipofuscin were also found.(ABSTRACT TRUNCATED AT 250 WORDS)


Ceroid lipofuscinosis in the border collie dog: retinal lesions in an animal model of juvenile Batten disease. Ceroid lipofuscinosis, an inherited disorder of lipopigment accumulation, was identified in a group of Border Collie dogs. The dogs developed mental, motor, and visual signs between age 15 and 22 months and progressed rapidly to severe neurological disease. The principal signs were blindness and gait and behavioural abnormalities with progressive dementia. Lipopigment accumulation was severe in neurones and glial cells of the central nervous system and was present in some visceral cells. Inclusions with variable ultrastructure were common in all cells of the retina, but the pigment accumulation did not damage the retinal architecture. The cytoplasmic inclusions were granular, sudanophilic, eosinophilic, and autofluorescent. Ultrastructural morphology varied, but fingerprint and curvilinear patterns predominated. The retinal lesions in the Border Collies were similar to those in English Setters with ceroid lipofuscinosis, but were much less severe than in juvenile human ceroid lipofuscinosis. This disorder bears a close resemblance to ceroid lipofuscinosis in English Setters and is another useful model for Batten's disease.


Anaesthesia and the 'inert' gases with special reference to xenon. Xenon has many of the properties of the ideal anaesthetic agent and has been proposed as a suitable replacement for nitrous oxide in routine clinical anaesthesia. Xenon, krypton and argon are chemically inert under most circumstances, yet all have anaesthetic properties. Xenon is of particular interest because it is the only 'inert' gas which is an anaesthetic under normobaric conditions. Because of this property, xenon has an important place in the history of the development of theories of anaesthetic action and of concepts such as MAC. Cost is likely to be a major impediment to the regular use of xenon.


Immunolabelling patterns of gap junction connexins in the developing and mature rat heart. The distribution of gap junctions in prenatal, postnatal, and adult rat hearts was studied by laser scanning confocal microscopy, using antiserum raised to a peptide (HJ) matching part of the sequence of connexin43 (a cardiac gap junction protein). Using digital reconstruction of optically-sectioned tissue volumes, a highly sensitive detection of immunolabelled gap junctions was achieved. The distribution of positive anti-HJ immunolabelling was regionalised in the prenatal heart from its first detection at 10 days post-coitus. High levels of immunopositive staining occurred in the trabeculae of the embryonic ventricles. Other zones of the early myocardium including early central conduction tissues had no detectable signal. The prenatal outflow tract, interventricular septum and a narrow zone of myocardium subjacent to the epicardial free wall also had low levels of immunopositive signal. During postnatal growth and in the adult rat heart, a marked distinction emerged between the central conducting tissues of the atria and ventricles. Whilst small immunostained gap junctions became detectable within the atrioventricular node on the atrial side of the junction, between the interatrial and interventricular septa, no immunolabelling was found within the ventricular branching bundle. This difference between the atrioventricular node and branching bundle is consistent with potential functional distinctions between these two structures, and is not consistent with the recent proposal that the His bundle and its branches act as an extended atrioventricular node in smaller mammals such as the rat. Ventricular Purkinje fibres, distal to the branching bundle, showed high levels of anti-HJ immunostaining. Organisation of gap junctions into intercalated disks within the ventricle proceeded late into intercalated disks within the ventricle proceeded late into the adolescent stages of heart growth. The distribution of a second connexin protein, MP70, not previously characterised in the heart, was studied using monoclonal antibodies. MP70 was transiently immunolabelled in the heart during the postnatal period, but only within valves. Previously, this protein has been reported only in the eye lens. MP70-containing gap junctions may represent a specialisation in avascular tissues, since blood vessels are not present in either the eye lens or the cusps of heart valves.


Isoflurane does not vasodilate rat thoracic aortic rings by endothelium-derived relaxing factor or other cyclic GMP-mediated mechanisms. Endothelium-derived relaxing factor (EDRF) is a potent endogenous vasodilator that has been indirectly suggested to play a role in isoflurane-mediated vasodilation. To examine directly the possible role of EDRF in isoflurane-mediated vasodilation, isolated rat thoracic aortic rings were suspended for isometric tension measurements, equilibrated to a resting tension of 2 g, and constricted with a 50% maximal concentration (EC50) dose of phenylephrine or KCl. Three groups of rings were studied: endothelium-intact, endothelium-denuded, and endothelium-intact rings treated with nitro-L-arginine methyl ester (L-NAME), a specific inhibitor of EDRF synthase. Isoflurane was then added at 1, 2, and 3% in a cumulative manner, allowing 10 min for each concentration to equilibrate. Indomethacin was present in all experiments to prevent the formation of vasoactive prostanoid metabolites. Since EDRF causes vascular relaxation by stimulating soluble guanylyl cyclase and increasing cyclic GMP, the effect of isoflurane on vascular ring cyclic GMP content was determined as an additional indicator of EDRF-mediated dilation. Rings with intact and denuded endothelium were isolated as described above, constricted with phenylephrine, and challenged with methacholine (positive control) or 1, 2, or 3% isoflurane. After 8 min exposure, the rings were flash-frozen in dry-ice-cooled acetone and homogenized in 1 N HCl for subsequent analysis of cyclic GMP content by radioimmunoassay. Isoflurane caused dose-dependent vasodilation of both KCl- and phenylephrine-constricted rings. In the phenylephrine group, at 2% and 3% isoflurane, endothelium-denuded and L-NAME-treated rings relaxed to a greater extent than endothelium-intact rings (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)


Spontaneous regression of juvenile nasopharyngeal angiofibroma. There is debate concerning the natural history of juvenile nasopharyngeal angiofibromas, especially whether or not they can spontaneously regress. Often claimed, spontaneous regression has not been well documented. To our knowledge, this is the first report in which a biopsy-proven juvenile nasopharyngeal angiofibroma spontaneously resolved. A second, less well-documented case is discussed.


Report on the first outbreaks of the porcine reproductive and respiratory syndrome (PRRS) in France. Diagnosis and viral isolation. We describe the first known occurrence in France of the porcine reproductive and respiratory syndrome (PRRS), a new porcine disease that first appeared in Germany in November 1990. Outbreaks of the disease appeared in November 1991 in Brittany (France), with comparable clinical features to those observed in other countries, were definitively confirmed by a serological analysis from the affected animals using an immunoperoxidase monolayer assay with PRRS-virus infected alveolar macrophages. Furthermore, we report the isolation from one serum of a filtrable agent that, in view of its cytopathic effect for porcine alveolar macrophages and of the serologic reactivity of the infected cells, appears similar to the Lelystad virus that has been implicated in the etiology of the disease.


An enzyme linked immunosorbent assay (ELISA) for the detection of antibodies to the porcine reproductive and respiratory syndrome (PRRS) virus. An indirect enzyme-linked immunosorbent assay (ELISA) has been developed for the rapid detection of antibodies to the porcine reproductive and respiratory syndrome (PRRS) virus in pig sera. Compared to the immunoperoxidase monolayer assay (IPMA) which was the only test available up till recently for serodiagnosis of the disease, the ELISA test proved to be more sensitive, particularly for early detection of antibodies. The test was also highly specific for the PRRS virus inasmuch as it scored negative all the hyperimmune sera directed to other swine viruses and the field sera sampled before the outbreak of the disease in Brittany (France). This easy test now provides further possibilities for epidemiological surveys and could also be a reliable tool for new sanitary prophylaxis of the disease.


[Laparotomy in HIV-infected patients. Indications, results apropos of 104 operated patients]. From May 1st 1983 to March 1st 1991, 126 laparotomies were performed on 104 patients infected by human immunodeficiency virus (HIV). We report the operative indications, macroscopic findings, anatomopathologic and microbiological results, and hospital mortality in this population. In 12.5% of cases, infection by HIV was established after operation. Sixty-eight percent of patients presented criteria for AIDS. The population was divided into 4 groups of indications: Group I: 30 patients had an emergency exploratory laparotomy. Laparotomy provided a diagnosis in 80% of cases, with a hospital mortality in 66%. Group II: 45 patients had an emergency laparotomy with a preoperative diagnosis. Hospital mortality was 24.4%. Group III: 29 patients had a non-urgent laparotomy with no mortality. Group IV: 14 patients were reoperated (22 laparotomies). For the entire population, hospital mortality was 9.3% for seropositive patients and 38.8% for AIDS patients.


[Cancer at confluence of the superior bile ducts with proliferating form. Nosologic, diagnostic and therapeutic aspects. Apropos of a case]. The authors report a case of fungating cholangiocarcinoma of the porta hepatis, revealed by jaundice in a 73-year- old cholecystectomized female. The diagnosis of obstruction of the hepatic duct junction suspected by ultrasonography and computed tomography was confirmed intraoperatively. Disobstruction was followed by laterolateral choledocoduodenostomy. An early recurrence required a second disobstruction with surgical then endoscopic insertion of a drain, with 23 months of satisfactory survival. The discussion is focused on hilar fungating cholangiocarcinoma. The extrahepatic (Klatskin tumor) or intrahepatic origin of these lesions, the circumstances and morphological investigations for the diagnosis are discussed. Particularities of the treatment are mentioned, with emphasis on the possible participation of the endoscopist for choledocho or hepaticoduodenostomy.


Maturation of mitochondrial and other isoenzymes of creatine kinase in skeletal muscle of preterm born infants. We studied pre- and postnatal changes in total creatine kinase (CK) activity, mitochondrial creatine kinase (Mi-CK) activity and immunochemical reactivity with anti-Mi-CK antibodies in skeletal muscle specimens from 12 infants, 10 of them preterm born, after a pregnancy varying between 28 and 40 weeks. Our results demonstrate that Mi-CK is present in fetal human quadriceps muscle and that the specific activity of Mi-CK increases during prenatal development from week 28 to 40 by a factor about two. Generally, adult levels have not been reached at birth, indicating a further postnatal increase of the activity of the enzyme. The Mi-CK protein content also increases during prenatal development. These results suggest that in human skeletal muscle the expression and accumulation of Mi-CK starts at mid-gestation, later than is known to occur for cytosolic CK.


The contribution of steroids to digoxin-like immunoreactivity in cord blood. Samples of cord serum from 29 healthy neonates were analysed for digoxin-like immunoreactive substance (DLIS), cortisol, 17 beta-oestradiol, progesterone, dehydroepiandrosterone-sulphate (DHEAS), 17 alpha-hydroxyprogesterone (17OHP), androstenedione, oestriol and ouabain-like activity (OLA; by inhibition of Na+, K+ATPase activity). The mean serum concentration of DLIS was 0.91 (SD = 0.19) nmol/L and the mean OLA was 26.1 (SD = 11.5) nmol/L. There was no correlation between DLIS and OLA. DLIS correlated significantly with oestriol (r = 0.521), progesterone (r = 0.534) and 17OHP (r = 0.43). Stepwise multiple regression analysis showed that 17 beta-oestradiol, progesterone and androstenedione contributed to DLIS and the intercept was 0.64 (SD = 0.127). The concentrations of steroids (17 beta-oestradiol, progesterone, androstenedione) required to displace digoxin by 50% in the digoxin immunoassay and inhibit Na+,K+ATPase in the OLA assay were 10(3)-10(4)-fold higher than those found in cord serum. We conclude that the contribution of these steroids to DLIS is small and that DLIS and OLA measure different compounds.


Beliefs about the satiating effect of bread with spread varying in macronutrient content. Beliefs about the satiating effect of foods varying in contents of proteins, fats, carbohydrates, and fibre were investigated by face-to-face interviews with a random telephone sample of 101 subjects. Respondents were presented with photographs of two slices of bread covered with common spreads. Fibre level was manipulated by using white bread or wholemeal bread. Levels of proteins, fats and carbohydrates were manipulated by using different kinds of spread. Low and high levels of proteins, fats and carbohydrates had similar energy contents. The results showed that protein-rich spreads (lean meat products) and high-fibre (wholemeal) bread were believed to have a strong satiating effect. The satiating effect of high-fat spreads (butter, bacon) was believed to be much smaller, whereas the sweet carbohydrate spreads (jam, honey) were believed to have no satiating effect.


Development of a lactococcal integration vector by using IS981 and a temperature-sensitive lactococcal replication region. A vector (pKMP10) capable of Campbell-like integration into the Lactococcus lactis subsp. lactis LM0230 chromosome via homologous recombination with chromosomal IS981 sequences was constructed from the replication region of lactococcal plasmid pSK11L, an internal fragment of IS981, and the erythromycin resistance gene and Escherichia coli replication origin of pVA891. The pSK11L replication region is temperature sensitive for maintenance in L. lactis subsp. lactis LM0230, resulting in loss of unintegrated pKMP10 during growth at greater than 37 degrees C. pKMP10 integrants made up 8 to 75% of LM0230(pKMP10) erythromycin-resistant cells following successive growth at 25 degrees C with selection, 39 degrees C without selection, and 39 degrees C with selection. pKMP10 integrants were also isolated from L. lactis subsp. lactis MG1363(pKMP10) but at a 10-fold-lower frequency (4%). No integrants were isolated form L. lactis subsp. lactis MMS368(pKMP10) (a Rec-deficient strain) or LM0230(pKMP1-E) (the corresponding plasmid lacking the IS981 fragment). Examination of 17 LM0230 integrants by Southern hybridization revealed pKMP10 integration into five different chromosomal sites. Four of the integration sites appeared to be chromosomal IS981 sequences, while one was an uncharacterized chromosomal sequence. The four IS981 integrants seemed to have pKMP10 integrated in a tandem repeat structure of undetermined length. Integrated pKMP10 was more stable (0 to 2% plasmid loss) than unintegrated pKMP10 (100% plasmid loss) when grown for 100 generations at 32 degrees C without selection.


IS946-mediated integration of heterologous DNA into the genome of Lactococcus lactis subsp. lactis. The lactococcal insertion sequence IS946 was used to construct suicide vectors for insertion of heterologous DNA into chromosomal and plasmid sequences of Lactococcus lactis subsp. lactis. Electroporation of L. lactis strains, including the recombination-deficient strain MMS362, with the suicide vector pTRK145 yielded 10(1) to 10(3) transformants per micrograms of DNA. pTRK145 insertions occurred primarily in the chromosome, with one insertion detected in a resident plasmid. Vector-specific probes identified junction fragments that varied among transformants, indicating random insertions of pTRK145.


Closure of fenestra in Clagett procedure: use of rectus abdominis musculocutaneous flap. Empyema developed in a 62-year-old man after right pneumonectomy for lung cancer. According to the Clagett procedure, an open window thoracostomy was made with two ribs removed. After 5 weeks, primary closure of the fenestra was attempted. Because the fenestra was too large to be primarily closed, a rectus abdominis musculocutaneous flap was successfully transposed to cover the chest wall. There was no evidence of recurrence of empyema during 11 months' observation after closure. In patients with a large fenestra and with little tissue left for closure, the rectus abdominis musculocutaneous flap could be of great help in completing the Clagett procedure.


Imaged thoracic lobectomy: should it be done? Imaged thoracic surgery is a new modality that is rapidly gaining acceptance from thoracic surgeons. Procedures that traditionally required a thoracotomy can now be done successfully using this technique in some patients. Three patients with primary carcinoma of the lung have undergone lobectomy using imaged thoracic surgery.


Receptor alterations in manganese intoxicated monkeys. The density of four different receptors and one marker of dopamine uptake sites were analyzed in monkey brains after manganese exposure (0.1 g manganese per month during 26 months, a dose comparable to that workers might inhale in dusty environments) by means of quantitative receptor autoradiography. The binding of 3H-mazindol to the dopamine uptake sites was reduced by 75% in both the head of the caudate nucleus and putamen, while it remained unchanged in the other regions analyzed. The binding of the D1 receptor ligand 3H-SCH 23,390 was reduced about 45% in the same areas as mazindol binding, while the density of D2 receptors was unaffected. The muscarinic acetylcholine receptors as well as GABAA receptors remained also unchanged in all brain areas analyzed after manganese exposure. Thus the dopaminergic neurons must be considered to be vulnerable to manganese concentrations attainable in the work environment. Our results also indicate that postsynaptic structures containing D1 receptors are sensitive while cells containing D2 receptors are either spared or compensated for by up-regulation of the number of receptors on remaining sites.


Calcium movement in vivo and in vitro in secretory-stage enamel of rat incisors. The lower incisors of young rats were dissected, immersed in physiological saline containing 45Ca under various conditions, and processed for autoradiography. The data were compared with those from in vivo 45Ca autoradiography. In secretory-stage enamel, wiped free of the enamel organ and immediately immersed in radioactive saline, there was intense labelling in the surface layers. The labelled area expanded only gradually into the deeper layers at a rate similar to that observed in vivo. Labelling in the enamel was similar in pattern but much weaker in intensity when the incisor was identically treated in vitro with the enamel organ attached. Glutaraldehyde pretreatment of the exposed enamel abolished expansion of the labelled area, whereas a hypochlorite pretreatment allowed a rapid diffusion of the isotope into the deeper layers of the secretory-stage enamel. The findings confirm the role of the enamel organ as a diffusion barrier to the penetration of calcium from the extracellular fluid to the secretory-stage enamel, and suggest an intimate correlation between physicochemical properties of the organic enamel matrix and the rate of surface-to-interior diffusion of calcium within the secretory-stage enamel of rat incisors.


Microwave tissue coagulator in liver resection for cirrhotic patients. The use of the microwave tissue coagulator was studied on 20 consecutive elective hepatic resections carried out for symptomatic hepatocellular carcinoma with liver cirrhosis. The mean operative blood loss (excluding one patient with hepatic vein injury) was 1132 mL. Five patients required no blood transfusion. The average time taken to coagulate the anticipated liver transection plane was less than 15 min. Apart from the complications similar to those occurring in hepatic resections for cirrhotic patients, higher incidences of intra-abdominal sepsis (20%), sympathetic pleural effusion in the absence of chest or intra-abdominal sepsis (20%), and persistent fever lasting more than 1 week (40%) were encountered. It was considered that these complications were related to the coagulated tissue present in the liver remnants (mean depth of tissue coagulation = 3.8 mm) and concluded that although the hospital mortality rate of 10% and the mean operative blood loss of 1132 mL were acceptably low, microwave liver surgery carried a high morbidity rate which is a drawback in major hepatic resectional surgery.


Turcot's syndrome.

Turcot's syndrome. A non-familial case of cerebellar medulloblastoma associated with adenomatous polyposis coli is described and the literature is reviewed. This is the second reported case of Turcot's syndrome in Australia. A greater understanding of the genetic implications of this rare condition might be achieved through reporting of cases and long-term documentation with polyposis registries.


Simultaneous use of rheoencephalography and electroencephalography for the monitoring of cerebral function. Many of the changes in human performance under stress or in hazardous environments may be attributed, in part, to altered cerebral functions. These changes may take place in either the cerebral neurologic or cerebral circulatory systems. Investigation of the interaction between intracranial blood flow and neural activity during these exposures would provide a better understanding of the human response to these conditions. Experimental and analytical procedures are described whereby the combination of rheoencephalography (REG) and electroencephalography (EEG) may provide additional information during the study of cerebral function. Examples are given to demonstrate the simultaneous use of these techniques during mental arithmetic and antiorthostatic bed rest. Results of these tests in both the time and frequency domains show that REG and EEG can be used to provide graded quantitative measures of cerebral responses to externally applied stressors.


Structure-function studies of the epidermal growth factor domains of human thrombomodulin. Structure-function relationships in the 6 epidermal growth factor-like domains of human thrombomodulin (TME, residues 227-462) were studied by deletion mutagenesis. Purified and characterised proteins were used for kinetic studies. Deletion of EGF1, EGF2 and residues 310-332 in EGF3 had no effect on thrombin binding (Kd) or on kcat/KM for protein C activation by the thrombin-thrombomodulin complex. Deletion of the rest of EGF3 and the interdomain loop between EGF3 and EGF4 had no effect on Kd but decreased kcat/KM to 10% of TME. Deletion of residues 447-462 of EGF6 had no effect on kcat/KM but increased Kd for thrombin approximately 6-fold. Thus, the region 333-350 in EGF3-4 is critical for protein C activation by the thrombin-thrombomodulin complex and the region 447-462 in EGF6 is critical for thrombin binding.


Membrane-altering effects of velnacrine and N-methylacridinium: relevance to tacrine and Alzheimer's disease. The interaction of pharmacological agents potentially useful in Alzheimer's disease, 9-amino-1,2,3,4-tetrahydroacridine (THA or tacrine) and its major metabolite velnacrine (or HP-029), along with related compounds with cytoskeletal proteins in human erythrocyte membrane was investigated using electron paramagnetic resonance spin labeling techniques. The results suggest that: (1) the position of the positive charge of tacrine may be important in the mechanism of its interaction with the membrane cytoskeleton; (2) like tacrine, velnacrine also strengthens cytoskeletal protein-protein interactions in erythrocyte membranes, but appears to be only about half as potent as tacrine. These results are discussed with relevance to therapeutic use of these agents in Alzheimer's disease.


Chemotactic LTB4 metabolites produced by hepatocytes in the presence of ethanol. Ethanol in low concentrations significantly alters the hepatocyte metabolism of the neutrophil chemotactic lipid leukotriene B4 (LTB4). Two novel metabolites of LTB4 which are encountered only when ethanol is present, retained significant biological activity. One metabolite, 3-hydroxy-LTB4 increased intracellular free calcium in the human neutrophil at concentrations as low as 3 x 10(-10) M as well as induced shape change and adherence to albumin-coated latex beads at 10 nM. The 3-hydroxy-LTB4 and 3,20-hydroxy-LTB4 metabolites were also potent chemotactic agonists with an ED50 at 3.0 and 9.0 nM, respectively. These results suggest that the presence of ethanol can substantially alter inactivation of LTB4 by the liver and may mediate neutrophil accumulation into the liver, thereby contributing to the pathogenesis of alcoholic hepatitis even when LTB4 biosynthesis occurs at some site distant to the liver.


Cigarette smoke exposure increases the cell water organization and membrane order of cultured T cells. In order to determine the effects of cigarette smoke (CS) exposure on the physical properties of cells, NMR water-proton relaxation time (which measures the intracellular water organization) and ESR spin labeling (which measures membrane order) measurements were performed on cultured Jurkat T cells exposed to CS. NMR spin-lattice relaxation time (T1) decreased with CS exposure in a dose-dependent fashion. A significantly depressed T1 value was obtained even when CS was delivered through a filter. Cell viability was not affected in this condition. Superoxide dismutase (SOD) prevented the depression of T1 value. These results suggest that superoxide radicals or subsequently generated species contained in the gas phase of CS increase the intracellular water organization in viable cells. CS exposure also increased the ESR membrane order parameter of nitroxide spin label. These physical characteristic changes may be important in CS-induced cell responses and cytopathology.


Antibodies directed against synthetic peptides distinguish between the catalytic subunits of protein phosphatases 1 and 2A. Two antipeptide antibodies (designated type 1 antibody and type 2A antibody) were raised against synthetic peptides, Cys-Thr-Pro-Pro-Arg-Asn-Ser-Ala-Lys-Ala-Lys-Lys and Cys-Val-Thr-Arg-Arg-Thr-Pro-Asp-Try-Phe-Leu, corresponding to the carboxyl termini of the catalytic subunits of protein phosphatase 1 and phosphatase 2A (Cys was added for specific coupling to carrier protein). These antipeptide antibodies were highly specific and were useful in discriminating between protein phosphatase 1 and phosphatase 2A in crude extracts or purified preparations. Type 2A antibody reacted with both native and denatured protein phosphatase 2A whereas under the same condition type 1 antibody reacted only with denatured protein phosphatase 1.


Enhancement of Ca2+-induced Ca2+ release in calpain treated rabbit skinned muscle fibers. Calpain treatment of rabbit skinned muscle fibers resulted in proteolysis of junctional foot protein or Ca2+ release channel of the sarcoplasmic reticulum. Electrophoretic and immunoblot analyses indicate that calpain cleaves off approximately 130 kDa peptide from the N-terminus. After such treatment, Ca2+ capacity of the sarcoplasmic reticulum remained normal and both Ca2+ and adenine nucleotide dependence of Ca2+-induced Ca2+ release mechanism were retained. However, the Ca2+-activated Ca2+ release rate was increased by two fold after the proteolysis. The results suggest the presence of functional domains in the junctional foot protein, and the N-terminus domain controls the activity of the Ca2+ channel without changing Ca2+ and nucleotide sensitivities.


Brefeldin A affects the cellular distribution of endocytic receptors differentially. The cell surface expression of three endocytic receptors was studied in human hepatoma Hep G2 cells treated with brefeldin A (BFA). Ligand binding and cell surface iodination revealed that BFA increased the number of mannose 6-phosphate/insulin-like growth factor II receptors twofold and decreased the amount of asialoglycoprotein and transferrin receptors by 40-60%. The altered expression of receptors at the cell surface was paralleled by changes in the respective ligand uptake. The implications of this finding on our understanding of intracellular trafficking are discussed.


A ring-reversed analog of atrial natriuretic peptide retains receptor binding, guanylate cyclase stimulation. We have prepared an atrial natriuretic peptide analog, ANP[13-27][1-12], in which the connectivity of the disulfide-linked ring has been reversed by formally cleaving the ring and cyclizing the N- and C-terminal tails. This analog, which retains many of the spatial relationships of the native molecule, binds to both ANP-A and ANP-C receptor subtypes, and triggers the production of cyclic-GMP by ANP-A. ANP-C binding of ANP[13-27][1- 12] is roughly equipotent to that of ANP itself, although the ring cleavage falls within the putative ANP-C binding domain. ANP[13-27][1-8], a truncated analog in which much of this binding domain has been removed, surprisingly maintains a high affinity for ANP-C; however, this peptide has lost the ability to activate the ANP-A-linked guanylate cyclase.


Regulation of flagellar motility by temperature-dependent phosphorylation of a 43 kDa axonemal protein in fowl spermatozoa. Phosphorylation of fowl sperm proteins was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) after incubating the demembranated spermatozoa with [gamma-32P]ATP at 30 degrees C or 40 degrees C. A marked difference of phosphorylation between 30 degrees C and 40 degrees C was observed in a 43 kDa protein. This protein was slightly phosphorylated at 40 degrees C, but strongly phosphorylated at 30 degrees C in a cAMP-independent manner. The motility of demembranated spermatozoa was negligible at 40 degrees C, but vigorous movement was observed at 30 degrees C. These results showed that phosphorylation of a 43 kDa protein is likely to be a regulatory step in the maintenance of fowl sperm motility.


Limited selectivity of a synthetic erbstatin derivative for tyrosine kinase and cell growth inhibition. The natural tyrosine kinase inhibitor erbstatin and a synthetic analog (1302) were co0pared for their inhibitory activity on EGF receptor kinase, PDGF receptor kinase and a src-type kinase from bovine brain. Erbstatin inhibited both growth factor receptor kinases equally well and had little effect on the src kinase. The analog exhibited similar potency for inhibition of purified EGF receptor tyrosine kinase as erbstatin, however, was clearly less effective for inhibition of purified PDGF receptor kinase as well as the src-type kinase. The selectivity was, however, not seen when the derivative was assayed with respect to inhibition of autophosphorylation of both growth factor receptors in Swiss 3T3 cell membranes. The latter finding might explain a lack of selectivity of the compound for inhibition of DNA synthesis in 3T3 cells when the cells were stimulated comparatively with EGF, insulin, EGF plus insulin or PDGF. The results suggest that the environment of growth factor receptors in the cell membrane can remarkably modify their susceptibility to tyrosine kinase inhibitors.


The protein kinase inhibitor, staurosporine, does not inhibit phosphoinositide kinases. The effects of the protein kinase inhibitor staurosporine on phosphatidylinositol (PI) kinase and phosphatidylinositol 4-phosphate (PIP) kinase activities in brain membranes, and the synthesis of polyphosphoinositides in cultured NIH 3T3 fibroblasts were examined. Staurosporine, at concentrations up to 1 microM did not significantly modulate either PI kinase or PIP kinase activity in brain membranes. DNA synthesis was dose-dependently inhibited by sub-nanomolar concentrations of staurosporine; however, cells treated with up to 1 microM staurosporine contained levels of PIP and phosphatidylinositol 4,5-bisphosphate (PIP2) similar to those observed in untreated cells. Therefore, phosphoinositide kinases do not appear to be either directly or indirectly affected by staurosporine.


Effect of chloride ions on the kinetic parameters of the potato tuber and mung bean pyrophosphate-dependent phosphofructokinase. The affinity constant Ka of PPi-PFK for Fru 2,6-P2 is equal to 1.56 nM for the potato enzyme and to 6.67 nM for that of the mung bean in the absence of chloride ions. These results are notably lower than the currently reported 5.5 nM and 30, 50 nM respectively. It is shown that the chloride ion is a competitive inhibitor of Fru 2,6-P2 for both enzymes. The inhibition constant Ki is equal to 15.6 mM for potato PPi-PFK up to 40 mM chloride. For the mung bean enzyme, the Ki is 19.0 mM up to 30 mM chloride. No effects are detected on the Michaelis-Menten constants Km of the substrates Fru-6-P and PPi up to 40 mM chloride. Other halide ions are also found to inhibit the potato PPi-PFK: bromide is competitive like chloride, whereas fluoride and iodide have a mixed inhibition towards Fru 2,6-P2.


Alterations induced by acylphosphatase in the activity of heart sarcolemma calcium pump. We studied the effect of muscle acylphosphatase on the Ca2+ pumping ATPase of heart sarcolemma. Acylphosphatase addition to calmodulin-depleted sarcolemmal vesicles produced a significant increase in the rate of Ca(2+)-dependent ATP hydrolysis, even higher than obtained with exogenously added calmodulin. Maximal stimulation (about four fold over basal value) was obtained with 550 units/mg vesicle protein, a concentration that fall within the physiological range. Conversely, similar amounts of acylphosphatase decreased the rate of ATP-dependent Ca2+ transport into the sarcolemmal vesicles. The maximal statistically significant inhibition of Ca2+ uptake was observed with the same acylphosphatase concentration that gave the maximal stimulation of Ca(2+)-ATPase activity. From these findings acylphosphatase appears to reduce the efficiency of heart sarcolemmal Ca2+ pump with an impairment of the coupling between ATP hydrolysis and Ca2+ transport. A possible mechanism of this effect is discussed.


Expression and assembly of Torpedo californica (Na,K)ATPase alpha-subunit truncated at N-terminal end in Xenopus oocytes. cDNAs for mutant alpha-subunits of Torpedo californica (Na,K)ATPase variously truncated at the N-terminal end were constructed and transcribed in vitro. Each of the mRNAs thus synthesized was co-injected into Xenopus oocytes together with mRNA for wild-type beta-subunit. Truncation of the alpha-subunit at trypsin accessible site T2(removal of the N-terminal 36 residues, alpha delta K37) led to a decrease in ouabain-sensitive ATPase activity and ouabain-binding capacity, leaving the amount of immunoprecipitable alpha-subunit unchanged. The Km values for Na+ and K+ of alpha delta K37 were about 10mM and 2mM, respectively, and fall in the same range for the wild-type ATPase. Truncation of the alpha-subunit leaving lysine-54(alpha delta K54) or alanine-79(alpha delta A79) resulted in the loss of the ATPase activity as well as a substantial decrease in the amount of immunoprecipitable alpha-subunit. Since the beta-subunit assembles with and thereby stabilizes the alpha-subunit, which is otherwise degraded rapidly, these results suggest that the segment of the alpha-subunit between lysine-37 and lysine-54 is involved in the assembly with the beta-subunit leading to the formation of the stable and active alpha beta complex.


Pyrimidine nucleoside monophosphate kinase isolated from pig brain homogenate catalyzes disproportionation of phosphate between two CDP molecules. An enzyme fraction that catalyzes CTP formation from CDP was purified from pig brain homogenate to a single band on SDS-PAGE. The preparation had a molecular weight of about 36,000 and showed a high activity of phosphorylating CMP and UMP by ATP and turned out to be one of pyrimidine nucleoside monophosphate kinases. It also catalyzes UTP formation from UDP, although rather weakly. These characteristics show that the enzyme species from pig brain homogenate has a rather broad specificity toward phosphate donor in phosphorylating nucleoside monophosphate.


Some biochemical changes in mouse after in-vivo irradiation of pancreas with ultrasound. 875 KHz continuous wave of ultrasound at 2.5 W/Cm2 intensity revealed certain biochemical and enzymological changes in mouse pancreas and liver following the irradiation of pancreas in-vivo for a total of 300 seconds spread over five days. The sacrifice of the animals was carried out on day 0, day 1, day 5 and day 10. Blood glucose was reduced significantly with concomitant increase in liver glycogen. Glucose-6-phosphatase in liver was decreased significantly while glycogen phosphorylase showed marginal variations. Increased calcium pool in pancreas along with Ca2+ activated ATPase was observed. These alterations were prevalent in all the days of sacrifice and also for more than 10 days after the last exposure. The results are suggestive of ultrasound could stimulate the release of pancreatic secretions.


Aldosterone-mediated Na+ transport in renal epithelia: time-course of induction of a potential regulatory component of the conductive Na+ channel. To correlate synthesis of aldosterone-induced proteins (AIPs) with the hormone's effect on transepithelial Na+ transport, protein synthesis in two model renal epithelia was examined during the natriferic response to aldosterone. Newly synthesized epithelial cell proteins were pulse-labelled with [35S]-methionine, separated by 2D-PAGE and detected by autoradiography. In both toad urinary bladder and A6 cells, a constellation of polypeptides which are a component of the renal epithelial conductive Na+ channel first appeared prior to, or concordant with, the increase in Na+ transport. The early synthesis of these proteins taken together with the fact that they appear to be the only aldosterone induced component of the multimeric Na+ channel, is compatible with the hypothesis that they are responsible for regulating Na+ channel activity during aldosterone-stimulated epithelial Na+ transport.


The identification of a membrane-bound myo-inositol 1-phosphatase in rat brain, liver, and testes. A membrane-bound myo-inositol 1-phosphatase has been solubilized and partially purified from rat tissues. This particulate enzyme was detected in brain, liver and testis and certain physicochemical and enzymological properties were examined. Previously this major enzyme of the inositol signaling system was considered strictly cytosolic. The ratio of activity in the membrane form was approximately one-eighth of the activity found with the cytosolic fraction. The molecular weight of this phosphatase was found to be 59,000 by gel filtration chromatography and a subunit molecular weight of 29,000 by Western blot analysis, values comparable to the cytosolic form. This phosphatase cleaves both D- and L- myo-inositol 1-phosphates which originate from two different cellular pathways and is inhibited by lithium ions. Polyclonal antibodies were raised against homogeneous testicular cytosolic myo-inositol 1-phosphatase and cross-reacted with this membrane form as determined by western blot analysis showing immunological identity.


Insulin alters the sensitivity of glycogen synthesis to inhibition by okadaic acid, a protein phosphatase inhibitor. We investigated the interactions of insulin and okadaic acid, an inhibitor of protein phosphatases type-2A and type-1, on glycogen synthesis in rat, guinea pig and rabbit hepatocytes. Insulin stimulated glycogen synthesis in rat and guinea pig but not in rabbit hepatocytes. In rat and guinea pig hepatocytes, the stimulation of glycogen synthesis by insulin was inhibited by low concentrations of okadaic acid (2.5-5 nM), which did not inhibit glycogen synthesis in the absence of insulin. In rabbit hepatocytes, insulin increased the sensitivity of glycogen synthesis to inhibition by low concentrations of okadaic acid even though it did not stimulate glycogen synthesis, and in the presence of insulin and okadaic acid (5 nM) glycogen synthesis was significantly lower than in the presence of okadaic acid alone. An increase in extracellular pH from 7.4 to 7.8 in a bicarbonate-free buffer, decreased the concentration of okadaic acid causing half-maximal inhibition of glycogen synthesis. It is suggested that an increase in cytosolic pH may be one mechanism by which insulin alters the sensitivity of glycogen synthesis to phosphatase inhibition.


Inhibition of 5-lipoxygenase and cyclo-oxygenase in leukocytes by feverfew. Involvement of sesquiterpene lactones and other components. Leaves or infusions of feverfew, Tanacetum parthenium, have long been used as a folk remedy for fever, arthritis and migraine, and derived products are widely available in U.K. health food shops. Previous reports have suggested interactions with arachidonate metabolism. Crude chloroform extracts of fresh feverfew leaves (rich in sesquiterpene lactones) and of commercially available powdered leaves (lactone-free) produced dose-dependent inhibition of the generation of thromboxane B2 (TXB2) and leukotriene B4 (LTB4) by ionophore- and chemoattractant-stimulated rat peritoneal leukocytes and human polymorphonuclear leukocytes. Approximate IC50 values were in the range 5-50 micrograms/mL, and inhibition of TXB2 and LTB4 occurred in parallel. Isolated lactones (parthenolide, epoxyartemorin) treated with cysteine (to neutralize reactive alpha-methylene butyrolactone functions of the sesquiterpenes). Inhibition of eicosanoid generation appeared to be irreversible but not time-dependent. We conclude that feverfew contains a complex mixture of sesquiterpene lactone and non-sesquiterpene lactone inhibitors of eicosanoid synthesis of high potency, and that these biochemical actions may be relevant to the claimed therapeutic actions of the herb.


Inhibitory effect of eugenol on non-enzymatic lipid peroxidation in rat liver mitochondria. The anti-peroxidative activity of eugenol on Fe(2+)-ascorbate- and Fe(2+)-H2O2-induced lipid peroxidation was studied using rat liver mitochondria. Eugenol inhibited thiobarbituric acid reactive substance (TBARS) formation induced by both the systems in addition to oxygen uptake and mitochondrial swelling induced by Fe(2+)-ascorbate. Time course studies on TBARS formation indicated the ability of eugenol to inhibit initiation and propagation reactions. There was no measurable chemical modification of eugenol during the course of mitochondrial peroxidation by both the systems. Mitochondrial peroxidation by Fe(2+)-H2O2 was inhibited by hydroxyl radical (OH) scavengers like mannitol, benzoate, formate and dimethyl sulfoxide apart from eugenol. The OH scavenging ability of eugenol was evident from its inhibitory effect on OH-mediated deoxyribose degradation. The second-order rate constant for the reaction of OH with eugenol was about 4.8 x 10(10) M-1 sec-1. Eugenol reduced Fe3+ ions and Fe3+ chelated to citrate or ADP but it did not exhibit pro-oxidant activity in OH-mediated deoxyribose degradation. Incubation of mitochondria with eugenol resulted in the uptake of small but significant quantities of eugenol which inhibited subsequent lipid peroxidation by acting as a chain breaking antioxidant.


Characterization of camptothecin-resistant Chinese hamster lung cells. Three camptothecin-resistant sublines (V79r, IRS-1r and IRS-2r) of V79 cells and their irradiation-sensitive mutants, IRS-1 and IRS-2, were developed by stepwise, continuous exposure to camptothecin (CPT). The degree of resistance varied among these cells. Based on the biochemical characterizations of these resistant cell lines, the mechanisms which could be responsible for the resistance to CPT were proposed to be: (a) a decrease in the intracellular accumulation of CPT with or without alteration of DNA topoisomerase I, (b) a decrease in the amount of DNA topoisomerase I, or (c) a decrease in the sensitivity of DNA topoisomerase I to CPT. The resistant cells which exhibited down-regulation of DNA topoisomerase I were collaterally sensitive to etoposide (VP-16) and its analogue, 4'-demethy-4 beta-(4"-fluoroanilino)-4-desoxypodophyllotoxin, despite the fact that there were equal amounts of DNA topoisomerase II in the parental and in the resistant cell lines. Alternating the usage of CPT and VP-16 for the treatment of cancer is indicated.


Inhibition kinetics and selectivity of the tyrosine kinase inhibitor erbstatin and a pyridone-based analogue. The inhibition mechanisms of the epidermal growth factor (EGF) receptor tyrosine kinase and the cAMP-dependent kinase activities by erbstatin and its analogue, RG 14921, were studied by kinetic analysis. Both compounds were slow-binding inhibitors of the EGF receptor kinase. Erbstatin inhibited the EGF receptor kinase as a partial competitive inhibitor with respect to both ATP and the peptide substrate, suggesting that it binds at a site distinct from the ATP and peptide binding sites of the enzyme, and thus lowers the binding affinities of the enzyme for both substrates. In contrast, the analogue RG 14921 inhibited EGF receptor kinase activity as a non-competitive inhibitor with respect to both ATP and the peptide substrate. The distinct modes of inhibition by structurally related compounds suggest a dynamic and possibly extended structure of the catalytic center of the kinase domain of the receptor. Erbstatin and RG 14921 exerted similar effects on cAMP-dependent protein kinase activity. In this system, both compounds displayed potent inhibition and acted by a mode of competitive inhibition with respect to ATP and non-competitive with the peptide substrate.


The effect of anion transport inhibitors and extracellular Cl- concentration on eosinophil respiratory burst activity. Furosemide has been shown recently to protect asthmatic patients against certain bronchoconstrictor challenges. We investigated the effect of furosemide on eosinophil function. Since furosemide may be exerting its inhibitory effect on the eosinophil by inhibiting anion transport, we also assessed the effects of the anion transport inhibitors 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB) and 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS). Furosemide, NPPB and DIDS inhibited the eosinophil respiratory burst in response to leukotriene B4 (LTB4) and, to a smaller extent, inhibited the response to opsonized zymosan (OZ). To assess whether the anion transport inhibitors were achieving their inhibitory effect by inhibiting an influx of Cl- ions into the eosinophil, the effect of removing extracellular Cl- on eosinophil function was determined. OZ-induced H2O2 production was inhibited by removing extracellular Cl- whereas the LTB4 response was not affected by the concentration of extracellular Cl-.


Disk diffusion sensitivity testing and antibacterial activity of azithromycin. Azithromycin (CAS 83905-01-5) disks with the selected loading (10, 15, 20 micrograms) were used for determination of the most suitable azithromycin disk concentration. Estimation was carried out by means of the regression line related to the zone size inhibition. Testing was performed on a variety of freshly isolated gram-positive, gram-negative and anaerobic bacteria derived from various specimens collected from patients. Using the disk diffusion method with 10 micrograms of azithromycin per disk in total 431 gram-positive, 875 gram-negative bacterial strains and 59 anaerobic bacteria were analysed. It was concluded that azithromycin disk containing 10 micrograms is sufficient for determination of bacterial sensitivity.


Further SAR in the new antitumor 1-amino-substituted gamma-carbolines and 5H-benzo[e]pyrido[4,3-b]indoles series. Using previously described techniques, various new 1-amino-substituted 5H-pyrido[4,3-b]indoles (gamma-carbolines, gamma-C) and 5H-benzo[e]pyrido[4,3-b]indoles (BPI) have been synthesized and evaluated. For known compounds containing a 1-[(dimethylamino)propyl] group, 1a and 1b in the gamma-C series and 2 in the BPI series are the most active. Studies with newly synthesized derivatives show that: (i) in the gamma-C series, the 4-unsubstituted-8-hydroxy-compound was inactive, whereas the 4-unsubstituted-9-hydroxy-5H-benzo[e]pyrido[4,3-b]indole is active; (ii) the 4-ethyl-8-hydroxy-5H-pyrido[4,3-b]indole derivative retains antitumor properties, but the 1-amino-substituted 4-ethyl-9-hydroxy-5H-benzo[e]pyrido[4,3-b]indole analog is devoid of biological activity; (iii) in the 5H-benzo[e]pyrido[4,3-b]indole series, the displacement of a hydroxyl group from the 9- to 10-position leads to inactive compounds. Based on the structural analogies, these results were unexpected because the same substituents on the 4-position lead to different biological properties in the two series.


Cellular regulation of the benzodiazepine/GABA receptor: arachidonic acid, calcium, and cerebral ischemia. The effects of cellular mediators that contribute to ischemia-induced neuronal degeneration on gamma-aminobutyric acid (GABAA)-receptor function were studied. In vitro, phospholipase A2 (PLA2) inhibited muscimol-induced 36Cl- uptake in cerebral cortical synaptoneurosomes. The major hydrolysis product of PLA2 activity, arachidonic acid, also inhibited GABA-mediated 36Cl- uptake. The unsaturated nature of arachidonic acid makes it (and its metabolites) highly susceptible to peroxidation by oxygen radicals. Incubation of synaptoneurosomes with the superoxide radical-generating system, xanthine and xanthine oxidase, decreased muscimol-induced 36Cl- uptake, suggesting that the peroxidation of arachidonic acid and/or its metabolites interferes with GABAA-receptor function. Another factor involved in ischemia-induced neuronal degeneration is an increase in intracellular Ca2+. Calcium also inhibited GABA-mediated 36Cl- flux, consistent with its ability to activate PLA2. In contrast, Mg2+, which blocks Ca2+ channels, enhanced muscimol-induced 36Cl- uptake, consistent with its neuroprotective effects. Each of these cellular processes is activated during cerebral ischemia and can lead to neuronal degeneration. We used a model of transient forebrain ischemia in gerbils to determine if GABAA-receptor regulation is altered in vivo at a time when CA1 hippocampal cells have degenerated. Four days after a 5 minute bilateral carotid artery occlusion, receptor autoradiography was performed to measure the binding of [35S]t-butylbicyclophosphorothionate (TBPS) to the GABA-gated chloride channel. Significant decreases in TBPS binding were observed only in the dendritic layers (stratum oriens and lacunosem moleculare) of the CA1 hippocampus. The results suggest that ischemia-induced cellular processes that contribute to cell death can decrease GABA-gated chloride channels on dendrites of CA1 pyramidal cells, and that GABAA receptors may also reside on neurons afferent to or intrinsic to the dendritic layers of CA1 hippocampus.


Limbic-hypothalamic-pituitary-adrenal axis activity and ventricular-to-brain ratio studies in affective illness and schizophrenia. Some investigators have speculated that structural brain alterations observed in some psychiatric patients might be related to increased limbic-hypothalamic-pituitary-adrenal axis (LHPA) activity. To explore this hypothesis, we prospectively studied 166 research volunteers (19 patients with research diagnostic criteria (RDC) major depression, 9 patients with RDC bipolar depression, 45 patients with RDC schizophrenia, and 94 RDC normal controls), examining the relationship between magnetic resonance image-determined ventricular-to-brain ratio (VBR) and indices of LHPA axis function (cerebrospinal fluid (CSF) corticotropin-releasing factor (CRF), CSF adrenocorticotropic hormone (ACTH), and 24-hour urinary-free cortisol secretion). We observed no significant differences in mean VBR among the three patient groups and the normal control volunteers. Of the indices of LHPA activity, only CSF CRF concentrations distinguished the four subject groups, with CSF CRF being significantly elevated in the more severely depressed major depression patients. Indices of LHPA activity were not significantly correlated with VBR in any of the three patient groups or in the normal volunteers. These preliminary results suggest that VBR is not highly associated with alterations in LHPA activity, at least as determined cross-sectionally. Further longitudinal studies with reference to diagnostic subtypes, severity, symptom profiles, and more specific neuroanatomic regions may allow the elucidation of possible relationships between LHPA pathology and structural brain alterations.


The role of bovine intraepithelial leukocyte-mediated cytotoxicity in enteric antiviral defense. The defense of a mucosal surface against viral infection is dependent in part on the leukocyte population resident at that site. In this study, leukocytes from the bovine intestinal epithelium were isolated and assessed for their ability to inhibit in vitro replication of an enteric pathogen, bovine coronavirus (BCV). As well, the intraepithelial leukocytes (IEL) were tested for their ability to mediate different types of cytotoxicity. The IEL were able to inhibit virus replication, and this activity was markedly enhanced by interleukin-2 and tumor necrosis factor. This combination of cytokines has similar effects on IEL-mediated cytotoxicity, which implicated cytotoxicity as a mechanism by which viral replication was limited. The IEL demonstrated enhanced cytotoxic function when compared with lymphocytes isolated from other sites in the gut-associated or systemic immune system. The IEL mediated higher levels of IL-2-activated, antibody-dependent, and lectin-dependent cytotoxicity than did lymphocytes from mesenteric lymph nodes, Peyer's patches, or the spleen. This function may be a reflection of the type of cell recruited to the epithelium, as indicated by the increased prevalence of T cells, and particularly CD8+ cells in the IEL population. Cytokine activation and the presence of a recognition signal, such as antibody, resulted in a synergistic increase in the level of IEL-mediated cytotoxicity. This type of interaction could serve to enhance the efficiency of IEL cytotoxic cells in vivo. Thus IEL-mediated cytotoxicity has the potential to serve as a mechanism of defense to enteric viral infection.


Differences in membrane order between C3H 10 T1/2 cells and their transformed counterparts as measured by EPR. Membrane order of mouse embryo fibroblasts and their ionizing radiation and chemically transformed counterparts was investigated using EPR spectroscopy after labeling the membrane of the cells with the fatty acid spin label, 5-nitroxy stearic acid. The EPR spectra were recorded at temperatures ranging from 18 to 38 degrees C for both control and transformed cells. The distance between the outer hyperfine splitting (2T' parallel), which is used as indicator of membrane order, varies in these two cell types. Below 28 degrees C. 2T' parallel is higher in transformed fibroblasts than in normal cells, whereas above this temperature membrane order is the same. Lipid analysis as carried out by the measurement of the cholesterol/membrane proteins and sphingomyelin/lecithin ratios, showed no difference in the amounts of the main membrane rigidifiers. These findings suggest that cell transformation of mouse fibroblasts induced by radiation or chemicals may produce alterations in the cell membrane, as evidenced by variations in its order at low temperature. These measured differences are presumably not attributable to its fatty acids composition but to its glycoproteins content, since changes in membrane rigidifiers were not observed between normal and transformed cells.


Correlation between levels of immunoglobulins and immune complexes in plasma of C57BL/6 and C57L/J mice infected with MAIDS retrovirus. Infection with helper-free, defective MAIDS murine leukemia virus (MuLV) caused a rapid polyclonal activation of B cells in 0.75-, 2-, and 6-month-old C57L/J mice (H-2b, Fv-1n/n), similar to that in C57BL/6 mice (H-2b, Fv-1b/b), which was recognized by elevated plasma immunoglobulin concentrations. However, changes in plasma immunoglobulin levels differed in C57BL/J and C57BL/6 mice. In C57L/J mice, infection resulted in a rapid increase in plasma IgM and IgG2a, and the elevation of IgG2a persisted undiminished for 21 weeks. Levels of IgG2b also became slightly elevated, but those of IgG1 and IgG3 were not significantly affected. Plasma of 6 to 7-month-old C57BL/6 mice contained already high levels of IgM (30-40 mg/ml), which persisted undiminished in uninfected mice but decreased progressively in infected mice to 10% of the original concentration during 25 weeks of observation. In C57BL/6 mice, plasma IgG1 and IgG2b as well as IgG2a became similarly elevated after infection but also only transiently. Their levels began to decrease progressively about 10 weeks after infection and fell to far below the maximum concentration observed. The drastic loss of plasma IgM and IgGs observed in C57BL/6 mice during the later stages of MAIDS MuLV infection did not seem to be a consequence of the polyclonal activation of B cells per se but seemed to reflect additional immunological abnormalities arising in infected C57BL/6 but not C57L/J mice. In both mouse strains these changes in plasma Ig levels correlated with the formation of Ig-containing immune complexes that bound to high-affinity, protein-binding ELISA plates in the absence of antigen coating, which may represent unusual forms of self-antigen-antibody complexes.


Recombinant baculovirus-expressed rotavirus protein (VP4) in an ELISPOT assay of antibody secretion. Studies on the protein specificity of the intestinal antibody response to rotavirus infection have been hampered by lack of antigenically conserved isolated proteins to serve as antigens in immunochemical assays. In this report, the use of an antigenically conserved baculovirus-expressed rotavirus protein (VP4) as a capture antigen in the ELISPOT assay is described. Anti-VP4 antibody-secreting hybridoma cells are used as a test population to show that expressed VP4 as the capture antigen detects numbers of antibody secreting cells comparable to intact rotavirus particles. Hybridoma cells specific for other rotavirus proteins are used to ensure the specificity of the expressed VP4 in the assay. The flexibility and ease of use of a recombinant expressed protein product as a capture antigen in this assay dramatically enhances the ability to quantitate intestinal antibody responses to specific viral proteins.