Prostate-specific antigen as a predictor of radiotherapy response and patterns of failure in localized prostate cancer. A study of preradiation and postradiation, serial serum prostate-specific antigen (PSA) levels was performed in patients who had clinically localized prostate cancer. The prognostic value of the PSA in pretreatment evaluation and posttreatment follow-up was assessed. Sixty-three patients who presented with clinically localized prostate cancer and who were treated with external-beam radiation therapy were followed-up for a median of 25 months. A serum PSA and physical examination were performed at 3-month intervals, and a bone scan was done yearly. An increase in PSA triggered an additional metastatic workup. Prostate rebiopsy was performed for new, palpable nodules or for a serial increase in PSA in the context of a negative metastatic workup. Forty-one patients remained recurrence-free and 22 recurred clinically, 15 distantly and seven locally. The PSA was the strongest, independent, pretreatment prognostic indicator (P = .019) among pretreatment PSA, stage, and grade, but lost significance when the serum prostatic acid phosphatase (PAP) status was included. The initial rate of the PSA decrease after radiation (median half-life, 2.6 months) failed to predict outcome. Recurrence-free patients reached postradiation PSA levels that were equivalent to those reported in disease-free male populations; failure of the PSA to reach such normal levels was a multivariate predictor of subsequent failure (P less than .037). All clinicopathologic documentations of failure were preceded by an increase in PSA levels during follow-up. Delayed versus early PSA increase was associated with clinically localized versus metastatic first recurrence. The serum PSA is an independent pretreatment and posttreatment predictor of outcome. Additionally, for a median follow-up of 25 months, delayed PSA failure is associated with clinically localized rather than metastatic recurrence, a relationship that may help in selection for local salvage therapy.


Samarium-153-EDTMP: pharmacokinetic, toxicity and pain response using an escalating dose schedule in treatment of metastatic bone cancer. Samarium-153 emits medium-energy beta particles and an imageable gamma photon with a physical half-life of 46.3 hr. When chelated to ethylenediaminetetramethylenephosphonic acid (EDTMP), it is remarkably stable in vitro and in vivo. In this study, we administered escalating amounts of 153Sm-EDTMP, from 0.1 to 1.0 mCi/kg (3.7-37 MBq/kg), to 22 patients with painful metastatic bone cancer. A complete concordance was found when the scintigrams of 153Sm-EDTMP were compared qualitatively to 99mTc-HDP bone images. Moreover, the skeletal uptake of the 153Sm-EDTMP related to the number of metastatic sites (r = 0.65; p = 0.001) showed an inverse proportion to the plasma radioactivity at 30 min following injection (r = -0.79; p = 0.0001) and was unaffected by the administered (mCi/kg), (r = 0.33; p = 0.13). Myelotoxicity was observed in 10 of the 29 treatment courses and leukopenia occurred in two. Thrombocytopenia occurred in patients who had low pretreatment platelet counts, albeit within the normal range (p = 0.001), most suffered from prostate cancer (p = 0.007) and retained a higher percentage of the 153Sm-EDTMP in their skeleton (p = 0.057). In four patients an exacerbation of the pre-existing pain ("flare reaction") was recorded. Pain palliation occurred in 65% of the treated patients (mean: 3.8 mo, range: 1-11 mo). Retreatment in first time responder patients was quite effective. Our preliminary results indicate that 153Sm-EDTMP is a promising radiotherapeutic agent for palliative treatment of metastatic bone cancer pain, and further study is necessary to ascertain its optimal dose, efficacy and toxicity.


Early response of canine temporomandibular joint tissues to arthroscopically guided neodymium: YAG laser wounds. A neodymium yttrium aluminum garnet (Nd:YAG) laser inserted through an operating arthroscope was used to introduce applied energy to synovial tissues, articular discs, and bone in the temporomandibular joint of mongrel dogs. Arthroscopic inspection of the wounds was performed at 1 and 2 weeks postoperatively. The animals were killed and the temporomandibular joints were examined grossly and histologically to determine the extent of injury and healing. The results show that laser wounds of bone and articular disc undergo no repair, whereas laser wounds of synovium show rapid repair. Laser damage to condylar marrow under an articular disc wound was unexpectedly found.


A neuroendocrine cause of oncogenic osteomalacia. All definite cases of oncogenic osteomalacia have, until now, been classified as mesenchymal tumours. We report here a case of oncogenic osteomalacia caused by a spinal tumour. Microscopically, it resembled the mixed connective tissue variant of previously described phosphaturic tumours. Immunohistochemical studies, however, showed the tumour cells to be positive for low molecular weight cytokeratin (CAM 5.2), S100 protein, PGP 9.5, and synaptophysin. Electron microscopy demonstrated neurosecretory granules. The histopathological findings strongly suggest that this is a neuroendocrine tumour.


A study of adhesion molecules as markers of progression in malignant melanoma. Adhesion molecules are substances which are involved in the interactions between cells, and between cells and the extracellular matrix in both benign and malignant tissues. Two members of this group--intercellular adhesion molecule-1 (ICAM-1) and MUC18--have previously been found to be expressed on melanoma; however, studies seeking a correlation between expression and metastatic behaviour have yielded conflicting results. In this study we investigated the expression of these two antigens and that of a number of other adhesion molecules [VCAM-1, ELAM, and the neural cell adhesion molecule (NCAM)] on a range of benign and malignant melanocytic lesions. Both ICAM-1 and MUC18 were found on a high percentage of all melanocytic lesions including benign naevi. VCAM-1 was found to be expressed on 79 per cent of benign naevi, 62 per cent of primary melanomas less than 1.5 mm in depth, and 6 per cent of thick primaries. The antigen was present on 14 per cent of lymph node metastases and on no extranodal deposits. This suggests that loss of melanoma cell adhesion mediated by VCAM-1 may be important in the development of metastatic melanoma.


Interstitial pneumonia in simian immunodeficiency virus infection. Interstitial pneumonia unrelated to Pneumocystis carinii or other infections was observed histopathologically in 5 of 25 rhesus monkeys infected with simian immunodeficiency virus (SIV). The predominant lesion was lymphocytic infiltration of interalveolar septa and hyperplasia of peribronchial and perivascular lymphoid tissue. Immunohistochemical staining using a panel of antibodies against human T and B lymphocytes, macrophages, and immunoglobulins showed that peribronchial aggregates and interstitial infiltrates were predominantly B cells, whereas perivascular masses consisted mainly of T cells. One animal with a primary B-cell lymphoma of the spinal cord had secondary plasmacytoid lymphomatous nodules throughout the lung which were accompanied locally by reactive B-cell lymphoid follicles. Another animal also had large areas of diffuse alveolar fibrosis and epithelial metaplasia to a bronchiolar type. In two monkeys, branches of the pulmonary arteries showed intimal proliferation and organizing occlusive thrombi, some of which were mineralized.


The Bologna Eubiosia Project: hospital-at-home care for advanced cancer patients. Since 1985 the Associazione Nazionale per lo Studio e la Cura dei Tumori Solidi (ANT), an Italian nonprofit anticancer organization, has provided a hospital-at-home care program for advanced and very advanced cancer patients. This program is part of the Eubiosia Project which also includes a number of complementary services. Teams of doctors, nurses, and psychologists work round-the-clock in the region and on hospital wards, assisted by diagnostic and therapeutic facilities applicable at home and by a round-the-clock on-call service seven days a week. The patients who were treated between December 1985 and December 1991 numbered 5603, of whom 2130 received treatment in 1991 alone, with 702 patients on line at the end of the period. The mean duration of home care for the first 2803 deceased patients was 62 days, and the percentage of deaths at home was 70%. We analyzed control of the main symptoms and the residual survival in three groups of patients--stomach, lung, and breast cancer sufferers--and found that acceptable pain control was achieved in 95% of cases. From the retrospective analysis of our data it can be seen that the eligibility criteria for entry to home care should not only depend upon performance status and the presence of symptoms, but should also take into account the factors that are inherent to the natural history of any kind of tumor as well as the social, domestic, and psychological characteristics of the patients.


Palliative care--could your patient have been managed at home? Palliative care, supportive care of the dying, is rapidly changing to better meet the needs of the patients and families. If palliative care is provided in the home rather than in hospital, there is a potential for improvement in the quality of life for patients and their families and a potential for cost reduction in the health care system. Our study was undertaken to determine whether or not palliative care patients admitted to University Hospital could have been cared for at home rather than in the hospital. The hospital charts of 96 palliative care patients were reviewed retrospectively. The results indicated that 61% of these palliative care patients did not receive any palliative care at home and that 94% died in an acute care hospital setting. Only 18% lived in a setting other than their own home, and 68% had a spouse or other family member living with them at the time of their final admission. Based on the level of support in the place of residence prior to final admission and the reasons for admission, we determined that many of the patients could have been managed at home for at least some of the palliative care period if appropriate support from a home care team had been available.


Palliative care needs of residents, families, and staff in long-term care facilities. While there is growing recognition that the physical needs of LTC residents have increased markedly in the 20 years, the palliative care needs of facility residents and their families are poorly understood. There also is a dearth of information on the educational and support needs of LTC facility staff vis-a-vis palliative care. Operating from the Juan de Fuca Hospital Society (a network of extended care facilities in Victoria), our Palliative Support Team (PST) was conceived in order to act as an educational resource to JdF staff as well as to provide expert consultation on palliative care issues. As part of the evaluation of this pilot program, a sample of 74 Juan de Fuca workers were interviewed to determine their perceptions of resident, family, and staff needs in relation to palliative care. In this paper, discussion is focused on the palliative care needs identified by professional caregivers. The ways in which a palliative care consultation team can address some of these needs are also discussed.


New perspectives in the treatment of decubitus ulcers. Although the consequences of prolonged lying on a hard surface are as old as the human race, publications on this subject are scarce. This is due to the fact that a decubitus ulcer never occurs in isolation but is nearly always a complication of some other condition. The term "decubitus" was already used by Hildnaus in 1590 and is derived from the Latin word decumbere which means "lying down". The most important fact to keep in mind is that pressure sores can be prevented. This is the reason why this article not only discusses the management of pressure sores but also the importance of preventive measures. The basis of effective treatment is early detection and an adequate knowledge of the fundamental pathological process. Only adequate preventive measures, careful examination of the lesions, and a thorough knowledge of the products used can avoid unnecessary suffering by patients.


The discovery of (2S,3S)-cis-2-(diphenylmethyl)-N-[(2-methoxyphenyl)methyl]-1- azabicyclo[2.2.2]-octan-3-amine as a novel, nonpeptide substance P antagonisst. We describe the structure-activity relationship development of a series of quinuclidines which culminated in the first potent, selective, nonpeptide substance P (SP) antagonist, (2S,3S)-cis-2-(diphenylmethyl)-N-[(2-methoxy-phenyl)methyl]-1- azabicyclo[2.2.2]octan-3-amine, 3 (CP-96,345). Compound 3 is a potent displacer of [3H]SP binding in human IM-9 cells and blocks SP-induced and capsaicin-induced plasma extravasation, as well as SP-induced salivation in the rat in vivo. This compound may both help to further our understanding of the interactions of small molecules with peptide receptors and serve to evaluate the therapeutic potential of a SP antagonist.


A hyperpolarization-activated inward current in human myocardial cells. Normally-polarized tissue from the human atrial myocardium usually exhibits a diastolic depolarization phase which can be suppressed reversibly by Cs+ or enhanced by inhibiting the inward rectifier K+ current, iK1, with Ba2+. (Escande et al., 1986). Because the suppression of the diastolic slope by Cs+ leads to a hyperpolarization of the cell membrane at the end of the diastolic phase, it was suggested that Cs+ might inhibit an inward current responsible for diastolic depolarization. Among the ionic mechanisms underlying the diastolic depolarization phase of cardiac tissues, the hyperpolarization-activated inward current, if, fits well to explain the small diastolic slope of human atrial fibres. In other preparations, this inward current carried both by Na+ and K+ ions is rapidly deactivated during the action potential and entirely blocked by millimolar concentrations of Cs+ (DiFrancesco 1981; DiFrancesco, et al., 1986; Kokubun et al., 1982; Callewaert et al., 1984; Denyer and Brown, 1990). Such a current in human myocardial cells has not been characterized so far although its existence in human atrial trabeculae was previously reported in an abstract (Carmeliet, 1984). In the present study, we describe an inward current which activates upon hyperpolarization in patch-clamped single human atrial cells and shares similar characteristics with the if pacemaker current described in unicellular and intact preparations of mammalian cardiac tissues.


Antiproliferative potencies of interferons on melanoma cell lines and xenografts: higher efficacy of interferon beta. Human melanomas have shown only limited responsiveness to clinical therapy with interferon (IFN). Our aim was to determine the most effective class of IFN for inhibiting growth of melanoma cells and to establish whether variation exists in response of various cell lines to different IFNs. We compared the direct antiproliferative effects of the type I IFN alpha-2b, IFN alpha-4a, and IFN-beta and the type II IFN-gamma on eight melanoma cell lines grown in vitro. We did this comparison by determining the concentration of each IFN that resulted in 50% growth inhibition, using the MTT [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium tetrazolium bromide] dye uptake method. We also tested IFN alpha-2a and IFN-beta for their ability to inhibit the growth of xenografts of the LiBr melanoma cell line in vivo in nude mice. Receptor binding was determined using [35S]methionine-labeled IFN alpha-4a, in competition with unlabeled IFN alpha-2b, IFN alpha-4a, and IFN-beta. The melanoma cell lines differed markedly in their sensitivity to the IFNs tested: Five were sensitive to low concentrations (less than 30 pM) of IFN-beta, only one was sensitive to similar concentrations of IFN alpha-2b, and none were sensitive to IFN alpha-4a at concentrations up to 920 pM. For all cell lines, the antiproliferative potency of the type I IFNs was IFN-beta greater than IFN alpha-2b greater than IFN alpha-4a. IFN-gamma was less active than IFN-beta on all except one of the cell lines. Similarly, IFN-beta was more potent than IFN alpha-2a in inhibiting the growth of the LiBr xenograft in nude mice. Labeled IFN alpha-4a bound with high specificity in all four melanoma lines tested, and competitive binding experiments showed that the order of binding affinity (IFN-beta greater than IFN alpha-2b greater than IFN alpha-4a) correlated with the order of antiproliferative potency. The finding that melanoma cell lines differ intrinsically in their sensitivity to IFNs may explain differences in clinical response. Our results suggest that IFN-beta may be the most effective IFN in the treatment of melanoma, although confirmation will require clinical trials involving large numbers of patients.


Transrectal microwave hyperthermia for benign prostatic hyperplasia: long-term clinical, pathological and ultrastructural patterns. Transrectal microwave hyperthermia of the prostate was administered to 191 patients with bladder outlet obstruction due to benign prostatic hyperplasia who were either at poor operative risk or who refused surgery. Patients were divided in 2 groups according to age and they underwent either 5 or 10, 60-minute sessions of hyperthermia, with a calculated intraprostatic temperature of 42.5 plus or minus 0.5C. Light and electron microscopy showed no irreversible damage at the glandular epithelium but did demonstrate a significant increase in neoformed intraprostatic capillary-like vessels. At 1, 12 and 24 months residual urine volume was significantly decreased in the majority of patients but only a minor amelioration of urinary flow rates and subjective symptoms was observed. According to maximum flow nomograms all patients were still obstructed postoperatively. Transrectal hyperthermia cannot be considered a genuine alternative to surgery for patients with bladder outlet obstruction due to benign prostatic hyperplasia.


Effect of Casodex on sleep-related erections in patients with advanced prostate cancer. Sleep-related erections in 5 patients with stage T3N0M0 prostate cancer treated with a new nonsteroidal antiandrogen, Casodex, were evaluated with continuous monitoring of penile tumescence and rigidity on multiple nights. Mean serum luteinizing hormone levels were 7.2 +/- 1.2 IU/l. before therapy and increased to 14 +/- 3.6 IU/l. after 6 months of therapy. Serum testosterone and estradiol levels also increased from a basal level of 5.05 +/- 1.9 ng./ml. and 102 +/- 18 pmol./l., respectively, to 8.04 +/- 1.32 ng./ml. and 175 +/- 20 pmol./l., respectively, after 6 months of therapy. No significant modifications in regard to number of nocturnal penile tumescence episodes, maximum penile circumference and total rigidity time were found before and after therapy. Only 1 patient reported a decrease in sexual drive and libido. All patients presented with stable disease (National Prostatic Cancer Treatment Group criteria) and an unmodified performance status (Eastern Cooperative Oncology Group) after 6 months. Pure antiandrogen therapy did not seem to interfere significantly with the erectile capability of men with prostate cancer.


Transrectal microwave hyperthermia for advanced prostate cancer: long-term clinical results. Transrectal microwave hyperthermia was applied to 46 stages D1 and D2 prostate cancer patients to treat urinary symptoms and local pain unrelieved by total androgen ablation therapy. Hyperthermia was administered in 10, 60-minute sessions twice a week for 5 weeks. A calculated intraprostatic temperature of 43.5 +/- 0.5C was maintained throughout the treatment. At 2 years the mean residual urine volume was significantly decreased (p less than 0.05), while the mean peak flow rate and maximum flow nomogram were improved but not significantly. The majority of patients reported a notable amelioration of subjective symptoms and quality of life. The only complication was a prostatorectal fistula that was cured by leaving a urethral catheter in place for 4 weeks. Prostatic hyperthermia is a safe and effective palliative procedure for bladder outlet obstruction due to advanced prostate cancer.


Quantitative morphometry of the adult human bladder. The primary objective of the present retrospective study was to characterize the effects of aging and BPH on bladder morphometry. Eighty-six bladder specimens were obtained from the autopsy archives of the Milwaukee County Medical Complex. The bladder specimens were divided into 4 groups based upon age and gender: Group I: males between the ages of 35-45 years; Group II: males between the ages of 65-75 years; Group III: females between the ages of 35-45 years; and Group IV: females between the ages of 65-75 years. The age groups were selected in order to identify a group of males with and without BPH. The area density of smooth muscle:connective tissue was determined in bladder specimens using color assisted computer image analysis. Masson-trichrome and double immunoenzymatic staining techniques were used to discriminate the smooth muscle and connective tissue elements of the bladder. The area density of smooth muscle:connective tissue in the Masson-trichrome stained sections was significantly greater in Group I vs. Group II (2.90 +/- 0.22 vs. 2.33 +/- 0.16) and in Group III vs. Group IV (2.85 +/- 0.13 vs. 2.03 +/- 0.20). Aging was associated with a decrease in the area density of smooth muscle:connective tissue ratio in both males and females. The area density of smooth muscle:connective tissue was not significantly different in younger males and females (Group I vs. Group III) and older males and females (Group II vs. Group IV). The present morphometric study suggests that aging and not BPH, is associated with a relative increase in detrusor fibrosis. Infravesical obstruction in BPH may effect bladder function via mechanisms unrelated to the histologic composition of the bladder.


Urinary tissue factor levels in transitional cell carcinoma of the bladder. Production of procoagulant activity by host and tumour cells may be increased in patients with cancer. Using a simple chromogenic assay, we have determined urinary tissue factor (TF) levels in patients presenting with transitional cell carcinoma of the bladder (TCC, n = 63), normal controls (n = 20) and patients with benign prostatic hypertrophy (BPH, n = 35). In addition, a separate cohort of patients undergoing endoscopic surveillance for superficial bladder cancer were studied to determine whether there was any difference in levels in those with recurrent disease compared to those with normal cystoscopies. Urinary TF activity was higher in TCC compared to controls (p less than 0.001) and patients with BPH (p less than 0.05). In patients undergoing check cystoscopy, those with recurrent disease (n = 32) had higher levels (p less than 0.01) than those with normal examinations (n = 21). It is concluded that urinary TF levels are elevated in bladder cancer and that this reflects disease activity in those at risk of recurrent superficial disease.


[Clinical evaluation of effects of KRN8601 (rhG-CSF) on neutropenia]. Clinical effects of KRN8601 (recombinant human granulocyte colony-stimulating factor:rhG-CSF) were studied in 26 patients with chronic neutropenia including 4 Kostmann's disease, 1 Shwachman's syndrome, 1 Lonsdale's syndrome, 1 glycogen storage disease Ib-associated, 6 chronic benign, 5 chronic hypoplastic, 2 cyclic, 4 autoimmune and 2 miscellaneous neutropenia. The patients were given rhG-CSF intravenously at doses of 20-540 micrograms/m2 or subcutaneously at doses 20-400 micrograms/m2, over the periods of 2-32 weeks. Increases in neutrophil counts occurred after rhG-CSF administration in 23 of the 26 patients. Patients with Kostmann's disease, Shwachman's syndrome and chronic hypoplastic neutropenia responded poorly compared to patients with other types of neutropenia. There were no serious side effects which caused interruption of the study. These results indicated a beneficial effect of KRN8601 in various types of chronic neutropenia.


[Immunocytochemical staining of decay-accelerating factor (DAF) on erythrocytes: paroxysmal nocturnal hemoglobinuria (PNH)]. Affected erythrocytes in patients with paroxysmal nocturnal hemoglobinuria (PNH) have been detected as complement-sensitive cells by the complement sensitivity assay. Decay-accelerating factor (DAF), a molecular mass of 70 kDa complement-regulatory membrane glycoprotein, has been reported to be deficient on affected PNH blood cells. In the present study, DAF on erythrocytes from 12 patients with PNH were stained by an immunocytochemical method and the ratio of DAF+ erythrocytes was compared with their laboratory data concerning hemolysis, almost all normal human erythrocytes stained positively for DAF. In contrast, various percentages of DAF+ erythrocytes were found in the patients with PNH. The percentages of DAF+ erythrocytes correlated positively with the total amounts of DAF on erythrocytes measured by an enzyme-linked immunosorbent assay (ELISA), negatively with % hemolysis in Ham's test and in sucrose hemolysis assay and with percentages of PNH type III (PNH-III) erythrocytes. In some patients with PNH, subpopulations of erythrocytes weakly-positive for DAF were demonstrated in addition to DAF- erythrocytes. The immunocytochemical method for staining DAF on erythrocytes developed in the present study is useful for the detection of affected PNH erythrocytes and applicable to further studies on membrane defects in PNH.


Alterations of anionic charge and/or sites of the glomerular basement membrane in the heterologous phase of passive Heymann nephritis. Alterations of the anionic charge and/or sites of the glomerular basement membrane (GBM) in the heterologous phase of passive Heymann nephritis (PHN) were studied. Rats with PHN induced by a single injection of anti-Fx1A IgG were examined at days 1, 2, 3 and 4. The left kidney was perfused with ruthenium red (RR) solution as a cationic probe. The RR particles (= anionic sites) in the GBM were counted and expressed as the number of RR particles per unit length of GBM. For quantitative determination of the total anionic charge of the GBM, the GBM-bound ruthenium (= anionic charge) was measured with an atomic absorption spectrophotometer (AAS). Abnormal proteinuria corresponding to a decrease in anionic charge was detected at days 3 and 4. The anionic sites in the lamina rara externa (LRE) adjacent to immune complex (IC) deposits were found to have diminished earlier from day 1 onwards. This diminution was largely confined to areas adjacent to the IC deposits and was significantly correlated with the amount of urinary albumin excretion. Proteinuria in the heterologous phase of PHN would thus appear to be causally related to a decrease in the number of anionic sites in the LRE adjacent to IC deposits.


Enzyme studies of methotrexate-resistant human leukemic cell (K562) subclones. Five methotrexate (MTX)-resistant K562 cell subclones (K562/MTX-1 approximately -5) were established and were examined for mechanisms of drug resistance. Impairment of MTX-polyglutamate formation, with membrane transport alteration, in the resistant cells was demonstrated in the previous studies (Koizumi, S. (1988) Jpn. J. Cancer Res. 79, 1230). Further analysis of sensitivity of the cells to trimetrexate (TMQ), which is not polyglutamated and does not require the reduced folate transporter, but is a potent inhibitor of human DHFR, revealed a modest decrease in sensitivity to TMQ (2.4- to 15-fold). Enzyme studies showed that the dihydrofolate reductase (DHFR) activities of these resistant subclones were very similar to that of the parent cells. The number of binding sites of these subclones for MTX calculated from Scatchard analysis was increased up to 7-fold in the K562/MTX-1 and -4 subclones and up to 3-fold in the other 3 subclones as compared to the parent cells. KD values of MTX for the DHFR in the K562/MTX-1 and -4 subclones also appeared to be altered relative to the parent cell line. Further, thymidylate synthase (TS) activity of the resistant subclones was reduced to 50% in K562/MTX-1 and -4 cells, and to 11-25% in the other subclones as compared to the parent cell line. These findings suggest that antifolate resistance in the newly established K562/MTX subclones in multifactorial with polyglutamation and transport defects accounting for the majority of resistance to MTX, and that alteration in the binding affinity of DHFR for MTX and diminished levels of TS may contribute to the 'residual' drug resistance to TMQ and have importance with respect to MTX.


Characterization by enriched polyclonal antibodies of developmentally regulated and cell type specific mouse testis antigens. Polyclonal antisera directed against testicular proteins were characterized by immunocytochemistry and Western blot analysis. Antibodies binding to testis-specific, developmentally regulated protein bands were eluted from their antigens and used for further characterization of the developmental profile and cell type-specific expression of two antigens, PSM33 and NNA75. While PSM33 was found to be present in spermatocytes from the late pachytene stage on, NNA75 could be localized in neonatal interstitial cells. NNA75 expression ceases by to postnatal day ten, when first meiosis starts within the seminiferous tubules, thus suggesting an interactive role of Leydig cells during the onset of meiotic divisions.


Synergistic induction of the differentiation of WEHI-3B D+ myelomonocytic leukemia cells by retinoic acid and granulocyte colony-stimulating factor. Retinoic acid (RA) has been shown to be an inducer of the terminal differentiation of several leukemia cell lines in vitro and in clinical trials to produce a high percentage of remissions in patients with acute promyelocytic leukemia. In an effort to increase the therapeutic efficacy of RA, we have measured the capacity of granulocyte colony-stimulating factor (G-CSF) to enhance the differentiation inducing activity of RA in WEHI-3B D+ monomyelocytic leukemia cells. Combinations of G-CSF and RA produced a supra-additive increase in the percentage of WEHI-3B D+ cells reducing nitro blue tetrazolium and expressing Mac-1 (CD11b) antigen on the cell surface, two markers of the mature state. In the presence of 50 ng/ml of G-CSF, which produced only 12% differentiation when used alone, 0.5 microM RA induced the same degree of cellular differentiation as the optimum concentration of RA (i.e. 7 microM) employed alone. The supra-additive differentiation produced by this combination was prevented by the presence of G-CSF monoclonal antibody in the culture medium, resulting in a degree of maturation comparable to that produced by the retinoid alone. When cells were sequentially exposed to G-CSF followed by RA, a much higher level of differentiation was obtained than when the order was reversed, suggesting that WEHI-3B D+ cells were primed to enter a differentiation pathway by G-CSF. The supra-additive terminal differentiation exhibited by the mixture of G-CSF and RA suggests that these agents should be evaluated for the therapeutic efficacy of the combination in patients with acute non-lymphocytic leukemia.


Mutagenesis of Escherichia coli: a method for determining mutagenic specificity by analysis of tRNA suppressors. A method for estimating mutagenic specificity in Escherichia coli (argE3, hisG4, thr-1, supE44), based upon the isolation of Arg+ or His+ revertants and identification of tRNA suppressors, is described. The method gives an insight not only into mutagenic pathways but also into the functioning of tRNA. With N-methyl-N'-nitro-N-nitrosoguanidine, 98% of mutations are GC----AT transitions. With N4-hydroxycytidine, 100% are AT----GC transitions. With hydroxylamine, apart from GC----AT transitions, approximately 30% of Arg+ revertants are formed by GC (or AT)----TA transversions. When the chemistry of the mutagenic attack is known, the method allows us to discriminate whether mutations occur on the transcribed or non-transcribed strands of DNA. It has been found that reversion of argE3 to Arg+ is a better monitor of mutagenic pathways than reversion of hisG4 to His+.


Characterization of two novel pre-B-cell lines (LK63 and LiLa-1): potential models of pre-B-cell differentiation. In this report we describe two newly isolated pre-B acute lymphoblastic leukaemia cell lines. Both cell lines lack EBV as detected by the EBNA-1 gene probed Southern-blots. Neither cell line expressed the B-cell-specific CD20 antigen on the cell membrane. However surface expression of CD20 was induced by phorbol ester (TPA) on both LiLa-1 and LK63 cell lines. Other pre-B and B-cell lines, such as Reh, Nalm-1, and BALL-1 did not exhibit these changes in phenotype. Previous immunoprecipitation studies have noted that a broad 50-55 kD band co-precipitates with the characteristic 33-37 kD CD20 protein. We demonstrate that, while the 33-37 kD CD20 species was undetectable on resting LiLa-1 and LK63 cells, in each case a 50-55 kD protein was immunoprecipitated by the CD20 antibody. However, the failure to detect any cell surface CD20-associated antigen on the control cells by immunophenotyping indicated that the CD20 epitope of the 50-55 kD molecule was not expressed on the cell surface. Following exposure to TPA the 50-55 kD species was reduced over 48-72 h while the level of the p33-37 CD20 protein was increased. Northern-blot analysis showed that the 50-55 kD protein was not a cryptic form of CD20 as the uninduced cells contained no detectable CD20 mRNA. The decrease of the 50-55 kD protein and the acquisition of the mature CD20 molecule were paralleled by a decline in proliferative activity in both cell lines. As expression of CD20 by normal pre-B cells also coincides with the cessation of cell division and maturation towards a mature B-cell phenotype, these cell lines appear to represent models for a discrete stage of B-cell differentiation which may be valuable in defining the signals regulating pre-B-cell proliferation.


Spermine and related polyamines produce a voltage-dependent reduction of N-methyl-D-aspartate receptor single-channel conductance. Several polyamines have been shown to interact with a site on the N-methyl-D-aspartate (NMDA) receptor that regulates the binding of open channel blockers. Spermine (SP) and spermidine (SD), polyamine agonists, enhanced binding of open channel blockers, whereas arcaine (ARC), diethylenetriamine (DET), and putrescine (PUT), polyamine antagonists, reduced the polyamine enhancement of open channel blocker binding. We previously showed that SP had multiple actions on NMDA receptor single-channel currents that underlie its effect on whole-cell NMDA receptor current. At high concentrations, SP produced a voltage-dependent decrease in NMDA receptor single-channel conductance and average open time. In the present study, another polyamine agonist (SD) produced a similar reduction of NMDA receptor single-channel conductance at higher concentrations. The polyamine antagonists (ARC, DET, and PUT), however, produced a voltage-dependent reduction in NMDA receptor whole-cell currents and reductions in single-channel conductance and average open time, even in the absence of polyamine agonists. The rank order of potency for reduction of NMDA receptor single-channel conductance by polyamines was ARC greater than SP greater than SD greater than PUT = DET, a rank order similar to that for the inhibitory actions of polyamines in receptor binding assays and for the effects of the antagonists on NMDA receptor whole-cell currents. The polyamine antagonist DET did not block the reduction of single-channel conductance by the polyamine agonist SP. In fact, the effects of SP and DET on single-channel conductance were additive. DET also showed a variable enhancement of NMDA receptor whole-cell currents in some neurons, suggesting polyamine agonist-like properties. These results are not consistent with the standard pharmacological profile for agonists and antagonists acting at the same site. Potential mechanisms for the effects of the polyamines on single-channel conductance are discussed.


Myeloblastoma formation in acute myeloid leukemia. The cell surface markers on the leukemic cells of 76 patients with adult acute myeloid leukemia (AML) have been analyzed by indirect immunofluorescence, and the presence of CD56+ leukemic cells was detected in ten of these patients. Four of these 10 CD56+ AML patients developed extramedullary myeloblastomas and in two of them an intracranial myeloblastoma. In contrast, in the remaining 66 CD56- AML patients, only one patient developed a myeloblastoma formation of the subcutaneous. It may be that the CD56 antigen which is an isoform of the neural cell adhesion molecule (NCAM), expressed on neurons, satellite cells of skeletal muscle cells, and on stromal cells, binds these tissues by a homophilic mechanism. CD56+ leukemic cells are capable of invading and of surviving in extramedullary tissues, where they proliferate and develop into a myeloblastoma. Because of this possibility, CD56+ AML patients should be carefully monitored for signs of myeloblastoma formation.


Characterization of bicuculline/baclofen-insensitive gamma-aminobutyric acid receptors expressed in Xenopus oocytes. I. Effects of Cl- channel inhibitors. Poly(A)+ RNA from bovine retina expressed gamma-aminobutyric acid (GABA)-activated membrane current responses in Xenopus oocytes, consisting of two pharmacologically distinct components. One component (IG-Aret) was mediated by GABAA receptors, and the other component (KG-BR) by atypical GABA receptors that were resistant to inhibition by bicuculline and insensitive to activation by baclofen. To further characterize the bicuculline/baclofen-insensitive GABA receptors, electrical recordings were made measuring the sensitivity of IG-BR to two Cl- channel inhibitors, t-butylbicyclophosphorothionate (TBPS) and picrotoxin. For purposes of comparison, effects of TBPS and picrotoxin were also assayed on currents mediated by GABAA receptors expressed in oocytes by rat cerebral cortex RNA (IG-Actx). The main finding of this study was that TBPS was a surprisingly weak inhibitor of IG-BR, whereas IG-Actx was potently suppressed. Assays on maximum responses indicated that IG-Actx was at least 500 times more sensitive to TBPS than was IG-BR (IC50 values of approximately 0.2 microM and greater than 50 microM, respectively). Moreover, inhibition of IG-Actx by micromolar concentrations of TBPS was largely insurmountable, whereas the weak inhibitory effects on IG-BR showed strong dependence on agonist concentration. For example, 10 microM TBPS reduced maximum IG-Actx by greater than 90%, an effect that was not significantly reversed by 10-fold increases in the concentration of agonist. In contrast, the same concentration of TBPS caused a 2-fold increase in the EC50 for IG-BR but had only marginal (less than 5%) inhibitory effects on maximum responses. Picrotoxin inhibited both types of current, but assays on maximum responses indicated that IG-Actx was approximately 30 times more sensitive than IG-BR (IC50 values of approximately 1 and 30 microM, respectively). Inhibitory effects of picrotoxin on IG-BR again showed strong dependence on agonist concentration, but in this case there was also a clear insurmountable component. Comparisons between IG-Actx and IG-Aret suggested that GABAA receptors expressed by either brain or retina RNA showed approximately the same sensitivity to TBPS and picrotoxin. Our experiments indicate that the bicuculline/baclofen-insensitive GABA receptors expressed by retina RNA differ markedly from GABAA receptors in their sensitivity to TBPS and picrotoxin. Defining the structural features responsible for these differences at the molecular level will provide a further means of investigating the complex mechanisms underlying interactions between inhibitors and GABA-activated Cl- channels.


Susceptibility of acute myelogenous leukemia blasts to lysis by lymphokine-activated killer (LAK) cells and its clinical relevance. To help understanding host-tumor relationships in acute myelogenous leukemia (AML) and better define indications for interleukin 2 (IL-2) therapy in this disease, we studied the relationship between the susceptibility of leukemic cells of 44 AML patients to lysis by autologous (26 cases) and/or allogeneic (41 cases) lymphokine-activated killer (LAK) cells and characteristics of the leukemia. Lymphocytes were activated in the presence of 1000 u/ml recombinant IL-2 for 5 days. Lysis of AML cells was studied by 51Cr release. Average lysis of AML cells by autologous LAK cells was 9 +/- 13% and by allogeneic LAK cells 10 +/- 9% with a significant correlation between lyses by both effectors (p = 0.01). Autologous (p = 0.005) and allogeneic (p = 0.004) lyses were higher in patients with initial infection. Allogeneic lysis was correlated with initial WBC count (p = 0.009), serum lactic-dehydrogenase level (p = 0.05), and expression of CD13 (p = 0.01). Autologous lysis was inversely correlated with expression of CD34 (p = 0.003). Expression of adhesion molecules CD54 (ICAM-1) and CD58 (LFA-3) by the leukemic cells did not correlate with their lysis by LAK cells. Susceptibility of leukemic cells to lysis by LAK cells did not correlate with prognosis of the leukemia.


Neutralization of bleomycin hydrolase by an epitope-specific antibody. Bleomycin hydrolase (BH) is a cysteine proteinase that terminates the pharmacological action of bleomycin (BLM). Amino acid sequence data obtained from a tryptic digest fragment of purified rabbit lung BH were used to synthesize a 14-amino acid peptide (LAVLEQEPIVLPAK; BHP14), which was conjugated to horseshoe crab hemocyanin and used to produce rabbit antiserum that was immunoreactive to both BHP14 and rabbit BH. Anti-BHP14 binding to BHP14 could be competitively blocked by the presence of either BHP14 or BH. Anti-BHP14 recognized both purified rabbit liver BH and postmicrosomal fraction from rabbit liver on Western blot, as a single band of M(r) approximately 48,000. Anti-BHP14 inhibited, in a concentration-dependent manner, BH activity in rabbit liver cytosolic fractions, as measured by deamido-BLM A2 formation. Thus, we have generated an epitope-specific neutralizing antibody to rabbit BH, which can block the metabolism of BLM by homogenates from rabbit tissue. These results suggest that the LAVLEQEPIVLPA epitope of rabbit BH is involved in the metabolism of BLM or is topologically near the active site. Furthermore, a BLM-resistant squamous carcinoma (C-10E) exhibited slightly more immunoreactivity, by enzyme-linked immunosorbent assay, to anti-BHP14 than did the parental A-253 cells, and a partially revertant (C-10E ND) cell line had intermediate anti-BHP14 binding. BH activity in these cell lines was in the same rank order as antibody binding, but differences in immunoreactivity were less than differences in enzymatic activity. Our epitope-specific neutralizing antibody should be useful in the further characterization of BH.


Epitope size, specificity and equilibrium constant for four monoclonal antibodies binding to the O:4 polysaccharide antigen of Salmonella serogroup B bacteria. One rat (MAST 83) and three mouse (MAST 107, 108 and 112) monoclonal antibodies (mAbs) directed against Salmonella serogroup BO lipopolysaccharide (LPS) were characterized and found to bind to the O:4 epitope but recognizing different surfaces of the polysaccharide chain. The epitopes were defined from the combined results of: (i) binding specificities in enzyme immuno assay (EIA) against chemically defined LPS and glycoconjugates; (ii) studies of affinity constants in Farr-assay for binding to oligosaccharides purified from LPS, or chemically synthesized; and (iii) knowledge of the conformation of the O-polysaccharide chain of Salmonella BO bacteria. Two of the antibodies, MAST 83 and 108, bound to the O:4 epitope when present in the terminal non-reducing end as well as an intrachain determinant of the O-polysaccharide, whereas MAST 107 and 112 bound only to the O:4 epitope when present as an intrachain determinant. The equilibrium constants (K values), determined for binding of the mAbs and a Fab-fragment isolated from one of them to a 125l-labelled tyramine-derivative of a Salmonella BO dodecasaccharide, were: 4.3 x 10(5) (MAST 83), 1.0 x 10(5) (MAST 107), 1.3 x 10(5) (MAST 107 Fab), 4.5 x 10(4) (MAST 108) and 1.9 x 10(5) l/mol (MAST 112). The results suggest that each epitope encompasses the O:4 specifying D-abequosyl residue together with different numbers of saccharides varying in size from di- to tetrasaccharides from the linear backbone chain. The antibodies also bind to different surfaces of the O-polysaccharide chain as suggested by its conformation.


Variable regions of antibodies to synthetic polypeptides--III. Antibodies arising in response to administration of anti-idiotope. Antibodies elicited by the synthetic polypeptide antigen (T,G)-A-L are directed against two distinct epitopes. The majority of antibodies bind to a GT containing epitope and bear an idiotope defined by monoclonal antibodies I7 and I9. In this study, we have examined the effect of in vivo administration of the I7 and I9 antibodies to mice. Administration of anti-idiotope elicits anti-(T,G)-A-L antibodies in all strains of mice tested, including genetic non-responders to (T,G)-A-L. These antibodies bind to GT and express the idiotope. Additionally, idiotope expressing antibodies that fail to bind to antigen are also produced. Monoclonal anti-(anti-idiotope) antibodies were made. One antibody bound to (T,G)-A-L, the other did not. Sequence analysis was performed and the V-regions of (T,G)-A-L binding antibodies were compared to those of the antibody that failed to bind antigen. Both sets of antibodies are derived from the same germline V-genes as the anti-(T,G)-A-L antibodies. These results have implications for understanding the nature of network regulation of the immune system and for those attempting idiotypic vaccination.


Hamster female protein binding to chromatin, histones and DNA. Hamster female protein (FP) is a member of the family of proteins known as pentraxins which share amino acid sequence homology, cyclic pentameric structure and calcium-dependent binding to ligands. Other members of this family include C-reactive protein (CRP) and serum amyloid P component (SAP), and most species synthesize both CRP and SAP. FP is unusual in that it is apparently the only pentraxin produced in hamsters, it is under hormonal control and it shares binding characteristics with both CRP and SAP. CRP has been defined and isolated by its calcium-dependent binding to pneumococcal C-polysaccharide via phosphocholine (PC) residues. SAP has been isolated by calcium-dependent binding to agarose. FP binds to both PC and agarose. Recently, both SAP and CRP have been found to bind to chromatin in a calcium-dependent manner and involvement of these proteins in the clearance of nuclear material has been proposed. In this paper we test whether FP shares the ability to bind to chromatin and histones, and compare its relative avidities for these ligands. Similar to CRP, FP bound to histones H1 and H2A, and chromatin. FP shared with SAP the ability to bind to DNA. However, FP binding was inhibited by PC for all ligands, whereas SAP binding was not. FP and SAP also failed to compete with each other for binding to DNA. By cross-inhibition FP bound much less well to PC than CRP, but was a very effective inhibitor of CRP binding to H2A. These studies demonstrate that chromatin and histone binding are conserved among these pentraxins. The role of the proposed PC binding site in these binding reactions is discussed.


Primary B-cell response to neuropeptide Y and bovine pancreatic polypeptide. An analysis of the murine primary response to protein epitopes has been made with two small highly structured proteins, neuropeptide Y (NPY) and bovine pancreatic polypeptide (BPP), both of 36-amino acid residue length and containing helical structures. A group of cell lines producing monoclonal IgM antibody have been prepared consisting of six anti-NPY and two anti-BPP. The VH nucleotide sequences have been determined and characterized as germ-line either by identity to established germ-line sequences or by inference from the germ-line character of the D and JH segments. The intrinsic association constants for the homologous ligands have been estimated to range from 10(4) to 10(7) M-1 based on competitive ELISA. No severe restriction in the utilization of VH families, D segments or JH segments appears to be involved in this response. Among the eight cell lines, three VH families were represented as well as all three families of D segments and all of the JH segments, although some preference for JH3 was indicated. The length of the N(D)N sequences was also not subject to restriction, ranging from 9 to 29. Two unusual features of the CRD3s were noted, one involving the utilization of an uncommon DSP2 segment and the other the apparent occurrence of a D-D fusion.


Immunodominant structures in the aminoterminal portion of human interferon alpha 1. The analysis of the antigenic structure of human interferon (IFN)-alpha 1 with a panel of monoclonal antibodies revealed four immunodominant regions. Three of them, recognized by 12 of 14 antibodies were mapped into the aminoterminal portion of IFN-alpha 1 around residues 31-38, 43-53 and 63-85. The region 31-85 proved important also for the antiviral and antiproliferative activity of the IFN-alpha 1 molecule. The antibody recognizing the sequence around residues (54)63-67 also inhibited the cellular binding of IFN-alpha 1 to the high-affinity receptors.


Immunochemical analysis of a snake venom phospholipase A2 neurotoxin, crotoxin, with monoclonal antibodies. Crotoxin is the major neurotoxic component of the venom of the South American rattlesnake, Crotalus durissus terrificus. The crotoxin molecule is composed of two subunits: a basic and weakly toxic phospholipase A2 (PLA2) called component-B (CB), and an acidic, nonenzymatic and nontoxic subunit called component-A (CA). Crotoxin exists as a mixture of several isoforms (or variants) resulting from the association of several subunit isoforms. We prepared monoclonal antibodies (MAbs) against each isolated subunit. Six anti-CA MAbs and eight anti-CB MAbs were tested for their cross-reactivities with each subunit and with other toxic and nontoxic PLA2s. Four of the six anti-CA MAbs cross-reacted with CB, whereas only one of the eight anti-CB MAbs cross-reacted with CA. Two anti-CB MAbs were found to cross-react with agkistrodotoxin, a single chain neurotoxic PLA2 purified from the venom of Agkistrodon blomhoffii brevicaudus. We determined the dissociation constants of each MAb for CA and CB isoforms and their capacities to neutralize the lethality and to inhibit the catalytic activity of crotoxin. We defined three epitopic regions on CA and four on CB, and used a schematic representation of the two subunits to characterize these epitopic regions with respect to: (1) the "toxic" and the "catalytic" sites of CB, and (2) the zone of interaction between the two subunits. We propose three-dimensional structures of the crotoxin subunits in which we localize amino acid residues that might be involved in the epitopic regions described here.


Immune recognition of a molecule naturally presented as a monomeric or an oligomeric structure: the model of the human chorionic gonadotropin alpha subunit. The immune recognition of a molecule naturally presented as a monomeric or an oligomeric structure is analyzed using the human chorionic gonadotropin alpha subunit (hCG-alpha) as a model. Indeed, hCG-alpha circulates as either a free subunit or combined to the beta subunit (hCG-beta) to form the dimeric hCG hormone. A T cell study was performed in BALB/c (H-2d) mice which were found to be high responders to hCG-alpha. Mice were immunized with the free hCG-alpha or the dimeric hCG alpha/beta, and their lymph node cells were challenged in vitro with either alpha subunits from different species, hCG or peptides spanning the entire primary structure of hCG-alpha. Proliferation and IL-2 assays demonstrated that hCG-alpha-primed lymph node cells responded equally well to hCG-alpha and hCG alpha/beta, suggesting that both the free and combined hCG-alpha subunits are processed in a similar way. Among the various synthetic peptides used, only those mimicking the hCG-alpha(59-92) C-terminus portion were able to stimulate hCG-alpha-primed lymph node cells, demonstrating that this region contains immunodominant T cell recognition site(s). The hCG-alpha(23-43) and (32-59) peptides, although incapable of stimulating T cells primed with hCG-alpha, elicited a T cell response when used as immunogens. These regions encompassed cryptic epitopes which were not generated during hCG-alpha processing in H-2d mice. The T cell epitopes of hCG-alpha above described as immunodominant or cryptic on the free alpha subunit, had similar characteristics when the alpha/beta dimer was used as the immunogen. In contrast, T cells primed with peptides mimicking immunodominant sites recognized differently the hCG-alpha and the hCG alpha/beta antigens. Moreover, the analysis of the B cell response to all the immunogenic hCG-alpha peptides indicated that they bear B and T cell epitopes as well. Antibodies elicited against the hCG-alpha(59-92) or (32-59) peptide were capable of recognizing the alpha subunit in its free form but not in the alpha/beta hCG dimer. Such study deserves attention for the comprehensive mechanisms of the immune response to hCG as well as for the design of anti-hCG vaccines.


Nitric oxide synthase activity in infantile hypertrophic pyloric stenosis. Hypertrophic pyloric stenosis is a common infantile disorder characterized by enlarged pyloric musculature and gastric-outlet obstruction. Its physiopathologic mechanism is not known, but a defect in pyloric relaxation (pylorospasm) has been postulated. Nitric oxide is a mediator of relaxation in the mammalian digestive tract, raising the possibility that pylorospasm could be caused by a defect in nitric oxide production. Since neuronal nitric oxide synthase and NADPH diaphorase are identical, we used the NADPH diaphorase histochemical reaction to study the distribution of nitric oxide synthase in pyloric tissue from patients with infantile hypertrophic pyloric stenosis. We studied pyloric tissue from nine infants with infantile hypertrophic pyloric stenosis and seven control infants and children. Cryostat sections were processed for NADPH diaphorase histochemical analysis. A polyclonal tau antiserum was used to identify the enteric nervous system by immunohistochemical methods. NADPH diaphorase activity was restricted to the enteric nervous system and blood vessels. In the pyloric tissues from the control patients, intense diaphorase activity was present in the nerve fibers of the circular musculature, in the neurons and nerve bundles of the myenteric plexus, and in some nerve fibers of the longitudinal musculature. In the pyloric tissues from patients with infantile hypertrophic pyloric stenosis, the enteric nerve fibers in the hypertrophied circular musculature were enlarged and distorted and did not contain diaphorase activity, whereas the activity in the myenteric plexus and the longitudinal musculature was preserved. We suggest that a lack of nitric oxide synthase in pyloric tissue is responsible for pylorospasm in infantile hypertrophic pyloric stenosis.


Analysis of C5b-8 binding sites in the C9 molecule using monoclonal antibodies: participation of two separate epitopes of C9 in C5b-8 binding. C5b-8 binding sites in C9 were examined using mAbs raised against C9. Among 16 mAbs, two, designated P40 and X197, blocked C9-mediated EAC1-8 lysis. C9 pretreated with the mAbs failed to bind to EAC1-8 at 4 degrees C. In addition, the mAbs became inaccessible to the C9 that had been incorporated into EAC1-8 at 4 degrees C. These findings suggest that C9 binding to EAC1-8, but not its membrane spanning or polymerization, is blocked by mAbs. By immunoblotting analysis using alpha-thrombin proteolytic fragments derived from C9 [a N-terminal fragment of mol. wt 25,000 (C9a) and a C-terminal one of mol. wt 37,000 (C9b)] and tryptic fragments of C9 (mol. wts 53,000 (C9a') and 20,000 (C9b')), the epitopes of P40 and X197 were mapped to the N-terminal and C-terminal regions of C9b, respectively. Both P40 and X197 bound to the C9 polymerized with Zn2+ in the fluid phase, whereas X197 but not P40 reacted with the membrane attack complex (MAC) formed on membranes. The results suggest that two distinct epitopes are involved in C9 binding to EAC1-8, and behave in a different manner for globular C9 bound to EAC1-8 at 4 degrees C, C9 assembled in MAC, or poly-C9 induced by Zn2+. These mAbs may be useful in clarifying the conformational states of C9 and in analyzing the molecular interaction between C9 and its inhibitors.


Examination of B lymphoid cell lines for membrane immunoglobulin-stimulated tyrosine phosphorylation and src-family tyrosine kinase mRNA expression. Crosslinking of membrane immunoglobulin (mIg) on B cells induces two signal transduction pathways: protein tyrosine phosphorylation and phosphoinositide turnover. A panel of murine and human B cell-lines, representing different stages of B cell development, was examined for the presence of anti-immunoglobulin-induced protein tyrosine phosphorylation. Of 10 B cell lines examined, only one, the human Raji cell line, had no detectably induced protein tyrosine phosphorylation. The pattern of proteins that were phosphorylated on tyrosine in response to mIg crosslinking differed somewhat in cell lines representing different stages of B cell development. Differences in the levels of constitutive phosphorylation of proteins were also observed between the cell lines. The identity of the tyrosine kinase(s) activated by membrane immunoglobulin ligation is not known. However, members of the src family of intracellular tyrosine kinases have been implicated as signal transduction molecules. As the tyrosine phosphorylation of proteins is a general phenomenon of signal transduction by membrane immunoglobulin, the tyrosine kinase(s) activated by it might be expected to be present in all cell lines in which the tyrosine phosphorylation signalling occurs. Therefore we examined these B cells for expression of mRNAs encoding the eight known src-like tyrosine kinases. Surprisingly, all eight kinase mRNAs were expressed in at least some of the B cell lines examined. The expression pattern of the fyn, hck, and lck genes suggests that expression of these kinases may be developmentally regulated in the B cell lineage. Three of the kinases, p55blk, p53/p56lyn and p60src, were detected in all 10 B cell lines. Whereas the src gene shows a ubiquitous pattern of expression, the expression of the blk and lyn genes is mostly restricted to cells of hematopoietic origin, and more especially B lymphoid cells. Thus, p55blk and p53/p56lyn may be particularly good candidates for the membrane immunoglobulin-activated tyrosine kinase.


Characterization of immobilized anti-CD3 antibody-activated T lymphocytes for use in adoptive immunotherapy of patients with brain tumors. Large numbers of T lymphocytes were successfully cultured for use in adoptive immunotherapy using immobilized anti-CD3 antibody. This method allows more than 1.5 x 10(10) T lymphocytes to be cultured within 2 weeks from only 20 ml of blood. Proliferated T lymphocytes were less cytotoxic than lymphokine-activated killer cells induced with a high dose of interleukin-2, but were more specific for autologous tumor cells as determined by cold target competition. Signal transduction of anti-CD3 antibody induced a population of CD8 positive cells, i.e. cytotoxic T lymphocytes. Adoptive immunotherapy using autologous lymphocytes requires a sufficient number of lymphocytes and high induced cytotoxic activity. This method is promising for clinical adoptive immunotherapy of patients with brain tumors.


Fine specificity of the murine antibody response to HIV-1 gp160 determined by synthetic peptides which define selected epitopes. In this report, we assess the humoral immune response in inbred strains of mice immunized with baculovirus-derived recombinant HIV-1 gp160 (rgp160). Six inbred strains of mice were each immunized with two different concns (5 and 50 micrograms) of rgp160, and the antibody response to rgp160 and synthetic peptides which define distinct gp160 epitopes was examined. Within a given inbred strain of mice, no significant difference in antibody titers to gp160 was observed in those groups receiving either 5 or 50 micrograms of rgp160 per injection. Following three immunizations with rgp160, differences in anti-gp160 titers were observed among the various inbred strains; however, these differences became less apparent after additional injections with rgp160. In addition, each mouse strain exhibited a unique reactivity pattern to seven gp160 epitopes defined by synthetic peptides. Multiple injections with rpg160 were required to induce responses to certain gp160 epitopes. The observed differences in the fine specificity of the humoral immune response to distinct gp160 epitopes among the six inbred strains suggest a genetic basis for regulating the antibody response to these epitopes. This apparent regulation can be overcome by multiple injections with rgp160.


Accessory middle cerebral artery and duplication of middle cerebral artery--terminology, incidence, vascular etiology, and developmental significance. A series of 455 bilateral carotid angiographies included 14 accessory middle cerebral arteries (Acc-MCAs) and seven duplication of middle cerebral arteries (Dup-MCAs). The branching patterns of Dup-MCA could be classified as "direct bifurcation" from the internal carotid artery, since most lacked the essential bifurcation or trifurcation at the distal end of the M1 portion. On the other hand, Acc-MCAs are probably residual congenital arteries. These anomalous MCAs were apparently associated with epilepsy. Five Acc-MCAs were associated with anterior communicating artery aneurysm at the origin. In addition, a rare case of Dup-MCA with arteriovenous malformation at its origin was found.


Prediction of motor function by magnetic brain stimulation in patients with intracerebral hematoma. Corticospinal motor pathways were monitored with motor evoked potentials (MEPs) elicited by transcranial magnetic stimulation in 13 patients with radiologically confirmed hypertensive intracerebral hemorrhage and varying degrees of hemiparesis. The electromyographic responses of the thenar muscles were recorded. The motor weakness of the upper extremity was assessed at initial monitoring and 3 months after hemorrhage, and correlated with changes in MEP. Absence of MEP in the acute stage indicated poor recovery of muscle strength. No false negative results were seen in our series. The presence of MEP in a completely hemiplegic patient predicted some recovery of motor function. The suppression of amplitude was more accurate than prolongation of latency in predicting the functional recovery. MEP monitoring of patients with hypertensive intracerebral hemorrhage in the acute stage can predict the outcome of motor function.


Intramedullary subependymoma with neurofibromatosis--report of two cases. Two cases of subependymoma in the cervical spinal cord associated with stigmata of neurofibromatosis are reported. Magnetic resonance (MR) imaging showed one tumor with a sharp margin, which was well-demarcated intraoperatively and was totally removed. MR imaging showed the other tumor with an irregular margin, which was partly invasive at operation. Subependymomas are rare in the spinal cord and these are the first reported associations with neurofibromatosis.


Characterization of T cell epitopes on the envelope glycoprotein of simian retrovirus 1 and 2 (SRV-1 and SRV-2) in several mouse strains. Various mouse strains were immunized with either SRV-1 or SRV-2 virus adsorbed on alum. Seven to 14 days later spleen cells were removed, and spleen cells were cultured with varying amounts of SRV-1 virus and SRV-2 virus, or varying amounts of selected SRV-1 and SRV-2 synthetic envelope peptides to determine their ability to initiate T cell proliferative responses. Our studies demonstrated that all mouse strains tested gave strong proliferative responses with SRV-2 virus. In contrast, SRV-1 virus induced T cell proliferative responses only in H-2k mouse strains. This apparent major histocompatibility complex (MHC)-restriction of SRV-1 virus-induced T cell proliferation correlates with the increased pathogenicity of SRV-1 virus in rhesus monkeys. The SRV envelope peptide 233-249 which is shared by both SRV-1 and SRV-2 virus initiates strong proliferative responses in both SRV-1 and SRV-2 virus immunized mice. The SRV-2 envelope peptide 96-102 initiates significant proliferative responses in SRV-2 immunized mice, and constitutes both a T and B cell epitope. The SRV-2 envelope peptide 127-152 has a 70% homology with the C-terminal region of SRV-1 peptide 142-167. The ability of SRV-2 peptide 127-152 to initiate T cell proliferation in SRV-1 virus immunized mice and the failure of the SRV-1 peptide 142-162 to initiate proliferation suggests that the region encompassing residues 160-167 must represent a T cell epitope in mice immunized with SRV-1 virus.


Intracranial seeding following surgery for spinal cord astrocytoma--case report. We report a case of intracranial dissemination developing approximately 6 months after partial removal of a spinal cord astrocytoma in a 40-year-old male. The clinical course and postmortem findings indicate that the tumor originated in the cervical cord and extended into the subarachnoid space, first the spinal canal and later intracranially. Spinal cord glioma dissemination through the cerebrospinal fluid is more common than previously considered and indicates a dismal prognosis. An aggressive approach, including radical surgery, entire neuraxis irradiation, and adjuvant chemotherapy, is suggested as the initial treatment for malignant spinal cord glioma to prevent subsequent dissemination.


Auto-antibodies to human sperm basic nuclear proteins in infertile and vasectomized men: characterization of antigens and epitopes recognized by antibodies. The sera of vasectomized men and of patients with immune infertility were used to study the antigens and epitopes of sperm nuclear proteins that bind antibodies in these sera. No reaction with sperm histones was observed except for one serum. P1, P2 protamines and pro-P2 protamines were recognized by auto-antibodies. Studies with peptides derived from P1 and P2 protamines and with mammalian protamines related to HP1 showed that antibodies are mainly specific for a folded protamine molecule, more especially antibodies from vasectomized men. These results disagree with the random coil model proposed for protamines by several previous works. A cross-reactivity between P1 and P2 protamines was observed only for the whole molecules and not for peptides derived from them. This observation suggests that the two classes of protamines, different in sequence, may have a similar folding and thereby may be functionally equivalent.


High cervical intradural lipoma--case report. A rare high cervical intradural subpial lipoma unassociated with spinal dysraphism manifested by a slowly progressive myelopathy simulating Brown-Sequard syndrome in a 37-year-old male. The diagnosis was based on neuroradiological imaging. Intraoperative recording of somatosensory evoked potentials showed recovery of the prolonged N20 latency, indicating adequate decompression was achieved.


Persistent trigeminal artery associated with brainstem infarct--case report. A 47-year-old male presented with a sudden onset of right hemiparesis and numbness of the left face. Magnetic resonance imaging demonstrated a lacunar infarct in the ventral pons. Cerebral angiography demonstrated a persistent trigeminal artery (PTA) anastomosing the left internal carotid artery to the distal basilar artery. Bilateral vertebral arteries and the basilar artery below the PTA junction were extremely hypoplastic. The bilateral posterior communicating arteries were embryonic. The posterior fossa circulation was almost independent from the circle of Willis. The poor vascular supply to the posterior fossa probably caused the brainstem infarct.


Coexistence of extracranial internal carotid artery aneurysm and multiple intracranial aneurysms--case report. A rare case of extracranial internal carotid artery (ICA) aneurysm coexisting with intracranial multiple aneurysms in a 64-year-old female is reported. The three intracranial aneurysms were clipped uneventfully by two-stage craniotomies. The extracranial ICA aneurysm at the infratemporal region was excised through a high cervical route and ICA was reconstructed by an end-to-end direct anastomosis. Ours is the first case reported of extra- and intracranial aneurysms surgically treated successfully.


Anomalous origin of the anterior cerebral artery and congenital skull dysplasia--case report. A 37-year-old male had a ruptured anterior communicating artery aneurysm associated with an anomalous right anterior cerebral artery originating from the right internal carotid artery immediately distal to the ophthalmic artery and running between the optic nerves. The anomaly was associated with congenital skull dysplasia and other systemic bone anomalies, apparently an incomplete form of cleidocranial dysostosis. The aneurysm was successfully clipped with hematoma evacuation.


Spontaneous epidural hematoma from a hepatocellular carcinoma metastasis to the skull--case report. A rare case of acute epidural hematoma originating from a hepatocellular carcinoma metastasis to the skull in a 52-year-old male is reported. The skull metastasis and epidural hematoma were completely removed, but he died of large liver tumor. Histological examination of the removed tumor showed many sinusoid-like blood vessels, which probably lead to hemorrhage and formation of epidural hematoma.


Effects of human, rat and porcine galanins on cardiac vagal action and blood pressure in the anaesthetised cat. Galanin (GAL) is distributed in sympathetic nerves in the cat, and exogenous GAL inhibits cardiac vagal action and lowers blood pressure in this species. This study on anaesthetised cats compares the effects on cardiac vagal action and blood pressure of human, rat and porcine GAL. Human GAL has only recently been sequenced. It is of particular interest as it is not C-terminally amidated, unlike porcine and rat GAL. Many regulatory peptides require a C-terminal amide group for their action. However, human GAL showed similar biological activity to the other (amidated) GALs here. Omission of a single amino acid (Ser6) from rat GAL significantly attenuated both cardiovascular actions studied here.


Immunologic identification of Na/Ca exchange protein in rat brain synaptic plasma membrane. Polyclonal antibodies raised against partially purified dog cardiac Na/Ca exchanger react with cardiac sarcolemmal proteins of 160, 120 and 70 kDa on SDS-PAGE. Using the same specific antiserum, we detected three prominent immunoreactive bands of about 150, 120 and 70 kDa on immunoblots with rat forebrain synaptic plasma membrane proteins. These data indicate that the Na/Ca exchange protein in rat brain synaptic plasma membrane is structurally and antigenically similar to the exchange protein in dog cardiac sarcolemma.


5-Hydroxytryptamine, substance P and thyrotropin-releasing hormone synapses in the intermediolateral cell column of the rat thoracic spinal cord. Serotonin-, substance P-, and thyrotropin-releasing hormone-immunoreactive profiles were studied in the intermediolateral cell column at the thoracic level of the rat spinal cord with light- and electron-microscopic immunocytochemistry. For each transmitter, a dense immunoreactive deposit was observed with the light microscope. At ultrastructural level, morphologically identified synapses amounted to 47% of all serotonergic varicosities, to 49% for substance P and 50% for thyrotropin-releasing hormone. Synapses appeared both symmetrical and asymmetrical. In each case, these synapses were mainly axodendritic (98%). These synaptic connections could mediate the physiological influence of these 3 substances in the spinal cord on the cardiovascular system.


Specificity and potency of N-methyl-D-aspartate glycine site antagonists and of mephenesin on the rat spinal cord in vitro. The potency, specificity and reversibility of various presumed glycine site N-methyl-D-aspartate (NMDA) antagonists was studied on neonatal rat spinal cord using the grease gap technique. 5,7-Dichlorokynurenate was the most potent and specific glycine site antagonist among the compounds tested. On the other hand mephenesin was a weak non-specific excitatory amino acid (EAA) antagonist; reduction of the response to NMDA was not reversed by D-serine. The EAA antagonist properties of mephenesin could explain its mode of action at the cellular level. The lack of effect of D-serine alone suggests that in our experimental conditions glycine sites on spinal neurones are occupied by an endogenous ligand.


Urinary concentrations of beta core fragment of hCG throughout pregnancy. We sought to determine a reference range for urinary immunoreactive beta core fragment of hCG (beta C-hCG) in pregnancy, the ratio between beta C-hCG and intact hCG, and the earliest detectable rise of beta C-hCG in urine. Urine was obtained from 741 pregnant women between 6-41 weeks' gestation, as well as from women undergoing donor insemination with timed ovulation peaks. The beta core fragment of hCG reached a maximum between 8-15 weeks, with a decrease between 20-29 weeks. The molar ratio of beta C-hCG to intact hCG was always greater than 1. In pregnancy, beta C-hCG concentrations increase in the urine in parallel to intact hCG but at a higher molar ratio, suggesting either placental production of beta C-hCG or enhanced metabolism of hCG to beta C-hCG in peripheral organs.


Corneal staining characteristics after sequential instillations of fluorescein. Twenty-one healthy noncontact lens-wearing subjects were screened for the presence of corneal staining with fluorescein after sequential instillations of the vital stain (Phase I). These staining characteristics were further explored by monitoring the results of sequential instillations of fluorescein at 2-h intervals over a 12-h period (Phase II) and by patching 1 eye overnight and observing corneal staining after sequential instillations of fluorescein in the morning after patch removal (Phase III). In Phase II more eyes stained in the early morning. Throughout the day the most common staining areas were inferior and nasal. In Phase III, 9 of 12 patched eyes presented with fluorescein staining, as did 7 of the 12 unpatched eyes. Corneal staining after sequential instillations of fluorescein is more likely to be observed early in the day and is not reduced by overnight patching.


[Successful treatment of hyperthyroidism simulating acute abdomen and psychosis]. A 49 years old female patient entered the surgical department because of epigastric and ileocoecal pains with the symptoms of acute abdomen. A surgical intervention was performed because of supposed appendicitis, but it was not verified. During the surgical observation the patient was confused and negativistic so she was transferred to the psychiatric department. Because of loss of 20 kg weight, high blood sedimentation and anaemia she was sent to our department with the suspicion of an organic disease. A moderate exophthalmos, glittering eyes and Graefe's sign was noted, therefore hyperthyroidism was diagnosed, which was proved by Kocher's blood picture, low serum cholesterol, extremely high T3 and T4 level, and iodine storage diagram. The antithyreotic treatment resulted a dramatic improvement in the extremely serious moreover hopeless case and after a long-term treatment the patient became symptom-free without complaints. Later because of regression of hyperthyreoidism and the growing nodular goitre the patient was treated on two occasions with radioactive iodine. At present the patient is in remission.


Decreased G-CSF and IL-3 production and gene expression from mononuclear cells of newborn infants. Newborns are predisposed to neutropenia and thrombocytopenia during bacterial sepsis. The presence of peripheral cytopenias during overwhelming infection may be secondary to decreased hematopoietic growth factor production during states of increased demand. We therefore examined circulating levels of granulocyte-colony stimulating factor (G-CSF) and IL-3, production of G-CSF and IL-3 from unstimulated and stimulated mononuclear cells (MNC), expression of G-CSF and IL-3 genes during unstimulated and stimulated conditions, and equilibrium and binding of G-CSF receptors on mature effector peripheral blood cells of adults and neonates. Serum from cord and adult peripheral blood contained negligible amounts of both G-CSF (less than or equal to 50 pg/mL) and IL-3 (less than or equal to 5 pg/mL). Constitutive supernatant levels of G-CSF and IL-3 from cord and adult unstimulated MNC were also undetectable. However, there was a significant difference in G-CSF and IL-3 production from stimulated cord and adult MNC. Supernatants from stimulated adult MNC had significantly more G-CSF (p less than 0.007) and IL-3 (p less than 0.02). Additionally, Northern blot hybridization and densitometry of autoradiographs demonstrated significantly more G-CSF and IL-3 mRNA transcripts from adult than from cord MNC. Lastly, affinity, binding, and number of G-CSF receptors on cord and adult peripheral effector cells were equal. These data suggest that, during states of increased demand, cord MNC produce less G-CSF and IL-3 than do adult MNC and have an associated reduction in their respective mRNA transcripts. These findings may have implications in the pathogenesis of neonatal cytopenias during states of increased demand, such as sepsis.


Selective impairment of taurine transport by cyclosporin A in a human placental cell line. We investigated, using a human placental choriocarcinoma cell line as a model, the effects of the immunosuppressive drug cyclosporin A on several placental transport systems mediating the transfer of glucose and amino acids from mother to fetus. The results of the investigation show that the transport system responsible for the transfer of taurine is selectively impaired by the drug, whereas the other transport systems are either stimulated or not affected. The inhibitory effect of the drug on taurine transport appears to be due to interference with calmodulin-dependent processes because calmodulin antagonists such as W-7, calmidazolium, and CGS 9343B mimic the effects of cyclosporin A. FK506, another immunosuppressive drug that is currently undergoing clinical trials, does not have this inhibitory effect.


Comparison of T cell functional changes during childhood with the ontogeny of CDw29 and CD45RA expression on CD4+ T cells. The ontogeny of the peripheral blood mononuclear cells' responsiveness to various activators during childhood was studied and compared to the expression of CDw29 and CD45RA molecules at the surface of CD4+ T cells. The results show that newborn peripheral blood mononuclear cells are characterized by a responsiveness to mitogens that is higher than that observed in adults, at least shortly after stimulation. This contrasts with a clear decreased response to CD2 and CD3 MAb at any time after stimulation. These functional characteristics correlate with a low density of CDw29 antigen on virtually all CD4+ T cells and a high density of CD45RA antigen on most CD4+ T cells at birth. These patterns of reactivity and phenotype are similar to those found among naive adult T cells. When ageing, the response to mitogens becomes rapidly similar to the adult's values, whereas the responses to CD2 or CD3 MAb are more gradually acquired. This slow rate of functional changes grossly parallels the increase of CDw29+ CD4+ and the decrease of CD45RA+ CD4+ T cell subsets. These changes finally lead to the immunophenotypic and functional characteristics that are typical of adult memory T cells. These results suggest that iterative antigenic stimulations both induce memory T cells and create the conditions to improve the overall immune competence.


[Computer-assisted speech training for aphasic patients--STACH and WEGE--in a self-help group]. Reported is our experience with two computer programmes for aphasic people (language loss after stroke). The STACH and WEGE programmes combine data banks of written language, pictures, and auditory speech output (delta modulation/Audicard 300 E by Speech Design, Munich) using MS-DOS. Special attention is given to the social environment, the patients' families and the context of self-help groups, who show great interest in this new facility for speech-training.


Human placental choline acetyltransferase activity at parturition. Choline acetyltransferase (ChAT) activity measured in human placentae obtained during labour, or after labour of spontaneous onset, was significantly lower than that measured in placentae obtained before the onset of labour. The reduction in ChAT activity may partly account for the reduction in placental acetylcholine (ACh) content and release into maternal vessels at this time. When cycloheximide (10 micrograms/ml) was infused for 3 h into both fetal and maternal vessels of the perfused placental lobule, it did not significantly alter placental ChAT activity or ACh output from the fetal vessels, though it significantly reduced placental beta HCG release from the maternal side of the perfused lobule. These results suggest that regulation of placental ChAT activity at the time of human parturition may be due to endogenous modulators of its activity, rather than to acute changes in the synthesis of this enzyme.


Intranephron distribution of glycine-amidinotransferase activity in rats. Guanidinoacetic acid (GAA), a precursor of creatine, is an essential substrate for muscle energy metabolism, and synthesized by glycine-amidinotransferase (transamidinase) mainly in the kidney. Since the intranephron distribution of transamidinase activity has never been quantified yet, the purpose of this study is to provide evidence about the localization of transamidinase activity using microdissected individual nephron segments. Synthesized GAA was separated by HPLC and detected fluorometrically after reacting with 9,10-phenanthrenequinone. Results obtained were as follows. (1) Transamidinase activity was distributed only in the first (S1) and the second (S2) portion of the proximal tubule, S1 being significantly higher than S2. (2) In S2, arginine and glycine were better substrates for GAA synthesis than canavanine and glycine. These results clearly indicate that GAA is synthesized in definite portions of the proximal tubule, and would be transported to the liver for further creatine production.


Tubular CO2 production from glutamine in the rat: segmental profile and modulation. The present study was designed to test whether tubular carbon dioxide production from the carbon skeleton of uniformly 14C-labelled glutamine exhibits quantitative and qualitative segmental heterogeneity. Our results show that CO2 production from glutamine in the proximal convoluted tubule (PCT) was dependent on substrate concentrations and is saturable at 10(-4) M of glutamine. Glutamine oxidation was demonstrable in all nephron segments examined. The PCT is the quantitatively predominant site of glutamine oxidation. Intermediate nephron segments, however, such as the thick ascending limb (MAL) and the distal convoluted tubule possess a significant capacity for glutamine oxidation, particularly when examined in terms of tubular protein content. Modulation of glutamine oxidation by extracellular pH was segment specific. Stimulation by acidosis and inhibition by alkalosis were observed in the PCT while carbon dioxide production from glutamine in the MAL was pH-insensitive. Glutamine oxidation was closely linked to sodium transport and greatly decreased by inhibition of Na-K-ATPase. In both the PCT and MAL, glutamine oxidation was inhibited by high extracellular potassium concentrations and in the PCT enhanced by extracellular hypokalemia. N-Ethyl maleiamide, an inhibitor of proton ATPase, led to almost complete cessation of CO2 production from the substrate in both PCT and MAL. Acetazolamide, an inhibitor of carbonic anhydrase, led to a partial reduction in carbon dioxide formation in the PCT, but did not affect glutamine oxidation in the MAL. We conclude that segmental qualitative heterogeneity characterizes oxidation of the carbon skeleton of glutamine with proximal segments showing the predictable effects of pH changes and carbonic anhydrase inhibition. The MAL appears to be nonmodulating.


[A case of apraxia and global aphasia without apraxia for axial body movement]. The case of a right handed 58 year old woman is presented who suffered an ischemic stroke after angiography following a vasospasm in the left internal carotid artery. The neuropsychological examination revealed global aphasia and severe apraxia for movements of the face and the extremities. However, the patient was able to carry out adequately axial movements to imitation and also to verbal command. These particular findings are discussed and explained within the context of the existing literature.


Effect of renal decapsulation on lithium excretion in the presence and absence of volume expansion. The relationship between fractional sodium excretion (FENa) and fractional lithium excretion (FELi) was determined in Munich-Wistar rats with intact capsules (control, n = 16), and in rats with acute bilateral renal decapsulation (n = 16) during hydropenia and acute saline volume expansion. In response to volume expansion, the glomerular filtration rate increased significantly in both decapsulated and intact groups, but was similar in the two groups of rats at the same period. The FENa and FELi increased significantly from 0.49 +/- 0.10 and 20.09 +/- 1.76% to 1.71 +/- 0.20 and 34.14 +/- 2.82% in control rats with volume expansion. In decapsulated rats, FENa and FELi were 0.17 +/- 0.03 and 11.64 +/- 1.39% during control and increased to 1.04 +/- 0.21 and 26.23 +/- 1.17% during volume expansion. The FELi and FENa were significantly greater in control rats compared with decapsulated rats during both control and volume expansion periods. The lower FELi in bilateral renal decapsulation suggests reduced delivery of sodium from the proximal tubule.


[Responsibility and guilt from the psychiatric/psychotherapeutic viewpoint]. The sense of universal responsibility of the individual is based on a religious (in a pre- and overconfessional sense) relationship to a transpersonal whole. It is a prerequisite for the philosophical discourse on ethics. The special responsibility of the psychiatrist in his anthropological constructs, his concepts of illness and disease, and his method of treatment are discussed. The patient's being responsible (at least partly) for falling ill, his coping with his disease and impairment, his cooperation during treatment and also the pathology of inadequate guilt feelings and deficits are dealt with. Taking the blame as part of the human condition may be seen as an indicator of a mature development of a person.


Isoproterenol infusion increases the maximal tubular capacity of phosphate reabsorption. Isoproterenol, a beta-adrenoreceptor agonist, decreases urinary phosphate (Pi) excretion; however, plasma phosphate concentration also decreases. The purpose of the present study was to determine the effect of isoproterenol infusion on phosphate reabsorption with concomitant phosphate infusions and in the presence and absence of parathyroid hormone (PTH). Clearance experiments were performed on male Sprague-Dawley rats which were acutely thyroparathyroidectomized (TPTX) and successive infusions of phosphate (1, 2, and 3 mumol/min) were used to determine the maximal tubular capacity of phosphate reabsorption (TmPi) factored for the glomerular filtration rate (GFR) in four groups of rats. In the saline-infused control group the TmPi/GFR was 2.87 +/- 0.19 mumol/ml (n = 8). When isoproterenol was infused intravenously at a rate of 0.005 mg/kg/min, urinary cAMP excretion was significantly increased and the TmPi/GFR was 3.53 +/- 0.17 mumol/ml (n = 10, p less than 0.05). In the PTH-infused group (33 U/kg bolus followed by a sustaining infusion of 1 U/kg/min) TmPi/GFR was 1.69 +/- 0.15 mumol/ml (n = 9). Coadministration of isoproterenol and PTH significantly increased the TmPi/GFR to 3.25 +/- 0.64 mumol/ml (n = 9). Basal cAMP excretion was similar in both groups. These results demonstrate that the stimulation of renal beta-adrenoreceptors by isoproterenol infusion markedly increases phosphate reabsorption and reverses the decrease in the maximal tubular capacity of phosphate reabsorption induced by PTH infusion.


Effects of uninephrectomy and high protein feeding on lithium-induced chronic renal failure in rats. Rats with lithium-induced nephropathy were subjected to high protein (HP) feeding, uninephrectomy (NX) or a combination of these, in an attempt to induce glomerular hyperfiltration and further progression of renal failure. Newborn female Wistar rats were fed a lithium-containing diet (50 mmol/kg) for 8 weeks and then randomized to normal diet, HP diet (40 vs. 19%), NX or HP+NX for another 8 weeks. Corresponding non-lithium pretreated groups were generated. When comparing all lithium treated versus non-lithium-treated groups, lithium caused a reduction in glomerular filtration rate (GFR) without significant changes in effective renal plasma flow (as determined by a marker secreted into the proximal tubules) or lithium clearance. Consequently, lithium pretreatment caused a fall in filtration fraction and an increase in fractional Li excretion. Lithium also caused proteinuria and systolic hypertension in absence of glomerulosclerosis. HP failed to accentuante progression of renal failure and in fact tended to increase GFR and decrease plasma creatinine levels in lithium pretreated rats. NX caused an additive deterioration in GFR which, however, was ameliorated by HP. NX+HP caused a further rise in blood pressure in Li-pretreated rats. The results indicate that Li-induced nephropathy, even when the GFR is only modestly reduced, is associated with proteinuria and arterial systolic hypertension. In this model of chronic renal failure the decline in GFR is not accompanied by a corresponding fall in effective renal plasma flow, which may be the functional expression of the formation of nonfiltrating atubular glomeruli. The fractional reabsorption of tubular fluid by the proximal tubules is reduced, leaving the distal delivery unchanged.(ABSTRACT TRUNCATED AT 250 WORDS)


Comparisons of models of cortical necrosis with segmental infarction. Sodium and water excretion were studied by standard clearance techniques in three experimental models where renal mass was reduced by superficial cortical necrosis (CN) or ischemic segmental infarction (SI). During hydropenia either CN or SI were able to conserve and regulate sodium to a very similar degree. After expansion of extracellular volume, CN reabsorbed less sodium and water than SI. In free-water clearance (CH2O) experiments, the 'apparent distal' sodium delivery was higher in CN than in SI, suggesting a decreased sodium and water reabsorption in the proximal tubules of juxtamedullary nephrons (JM). Both kidneys had similar CH2O when factored for inulin clearance but when CH2O was corrected for 'apparent distal' sodium delivery it was lower in CN than in SI, demonstrating an incapacity of JM to dilute urine. CN also showed less capacity to reabsorb free water than SI. Thus, the use of CN and SI within the same animal was useful to study functional differences between superficial and juxtamedullary nephrons. The present study also suggests that the kidney with superficial CN was unable to perform maximal urine dilution and concentration.


[The first interview and construction of the psychodynamic hypothesis]. The development of psychoanalitic short term therapy confirms the importance of the first interview for estimating the expectations of the patient and elaborating the first hypothesis. The therapist is immediately caught in the dynamics of pretransference and precountertransference. He depends upon the comprehension of his own emotional reactions to understand the actual affection and the nodal conflict. The psychodynamic hypothesis should be inferred from what both the patient and the therapist are saying and acting.


[Recalling the acute psychotic experience]. In this study we have analysed the memory of the acute psychotic experience in chronic schizophrenics to evaluate the influence of memory on present symptomatology. We briefly examine the literature on this subject, then we describe some clinical cases trying to give a sense to the different way of "recovery", in schizophrenics. In particular we note that sensorial events of the acute psychotic experience have an important role in memory. Besides this study deepens some particular aspects of memory in schizophrenics.


Gold labelling of RNA in virus-induced mitochondrial vesicles in the sheep blowfly Lucilia cuprina. Membranous vesicles are thought to be the replication site for viral RNA of many plant and animal viruses. A relatively rare site of virus-associated vesiculation is that of the mitochondrial outer membrane. In this study, virus-induced mitochondrial vesicles of the blowfly, Lucilia cuprina, were labelled with ribonuclease/gold and an antibody against double stranded RNA (anti-polyinosinic:polycytidylic acid). Both methods showed the presence of RNA in the vesicles thus indicating they may be a site for viral RNA replication in Lucilia.


Endothelium-derived nitric oxide synthase inhibition. Effects on cerebral blood flow, pial artery diameter, and vascular morphology in rats. We determined the effects of inhibiting the production of cerebral endothelium-derived nitric oxide on pial artery diameter, cortical blood flow, and vascular morphology. An inhibitor of endothelium-derived nitric oxide synthesis, NG-nitro-L-arginine methyl ester hydrochloride (L-NAME), or an equivalent volume of 0.9% saline was infused into rats intra-arterially in a retrograde fashion via the right external carotid artery at a rate of 3 mg/kg/min to a total dose of 190 mg/kg or intravenously at 1 mg/kg/min to a total dose of 15 mg/kg. Large pial arteries were continuously visualized through an operating microscope, and cortical cerebral blood flow was monitored by laser-Doppler flowmetry. To localize areas of morphological interest, the protein tracer horseradish peroxidase was injected 15 minutes before termination of the L-NAME infusion and the rats were perfusion-fixed 15 minutes later for light and electron microscopic analysis. Infusion of L-NAME significantly raised arterial blood pressure at both doses (for 190 mg/kg, from 103.2 +/- 3.4 to 135 +/- 3.4 mm Hg; for 15 mg/kg, from 125 +/- 2.8 to 144.4 +/- 4.0 mm Hg). Pial arteries constricted within 10 minutes after the start of the intracarotid infusion to 40% of the preinfusion diameter, while cortical cerebral blood flow decreased to an average of 72.5% of that at baseline. Morphological abnormalities in the experimental rats included microvascular stasis and focal areas of blood-brain barrier disruption to protein. Ultrastructural examination of cortical leaky sites revealed constricted arterioles with many endothelial pinocytotic vesicles and microvilli. These observations suggest that inhibition of endothelium-derived nitric oxide synthesis affects the relation between cerebral arterial diameter and cerebral blood flow and can lead to subtle cerebral vascular pathological changes consistent with focal brain ischemia.


Immunotherapy for superficial bladder cancer. A developmental and clinical overview. Superficial bladder cancer is one of the few solid human malignancies in which immunotherapy has been proved to be effective. Bacillus Calmette-Guérin was the vaccine which opened the door for this innovative approach. In an era of remarkable achievements in biotechnology, it is truly amazing that this throwback to the Stone Age of tumor immunology has not yet been replaced by a more (or equally) effective substitute. Potential candidates are already on the horizon and deserve a comprehensive evaluation. They must show not only that they are devoid of significant adverse effects but that they possess, beyond a doubt, superior antineoplastic activity. Even more remarkable is that one of the oldest vaccines still in use could emerge in a new role as an effective antineoplastic agent. Bacillus Calmette-Guérin has demonstrated an uncanny capacity for effectiveness as therapy for human diseases. Its protective effect against tuberculosis is well recognized, and its contribution to cancer therapy is widely known. A new and increasing repertoire has recently been presented: two separate groups of researchers have employed the vaccine successfully as a vehicle to express antigen-encoded genes from other pathogens. The exciting aspect of these recent studies resides in the demonstration that the altered vaccine is able to induce humoral and cell-mediated immune responses to the recombinant antigens, including the human immunodeficiency virus (HIV). Thus, BCG once more attracts enormous interest from the scientific community for its versatility and potential as a therapeutic agent.