Search

Forty-five patients with 63 bladder tumors were treated with transurethral neodymium: YAG laser irradiation. The operation was performed under lumbar or local anesthesia, the bladder being irrigated with a 10 per cent urigal solution. A five-second beam of 50 watt electric power was chosen as one unit of irradiation, and 5 to 50 units of irradiation were utilized for one operation. Forty-one tumors were completely resected with laser irradiation alone. In the other 22 tumors, electroresection or partial cystectomy was undertaken after the laser irradiation. Pronounced complications did not develop in any of the patients. Perforation was not experienced in our series. The bleeding at irradiation was minimal. Laser surgery through endoscopy thus represents a promising approach for the treatment of bladder tumors.

A cytochemical technique for demonstration of neomycin binding sites by electron microscopy was developed and applied to Escherichia coli. Neomycin was conjugated chemically with bovine serum albumin (BSA). Colloidal gold was coated with the conjugated neomycin-BSA. The neomycin-BSA-gold was applied to thin sections of Epon-embedded E. coli and examined. Gold particles were observed on the outer membrane and the cytoplasmic membrane of E. coli. It was probably the ribosomes that were being labeled in the cytoplasm. Different cytochemical controls, including a number of inhibition tests and the use of BSA-gold, proved the specificity of this cytochemical technique and provided the biochemical significance of the observations.

A concanavalin A (Con A)-mediated polyclonal helper assay system was established by using the thymus cells or splenic T cells as a source of helper T cells. When splenic B cells were cocultured with thymus cells or splenic Lyt-2- T cells in the presence of an optimal dose of Con A, B Cells were polyclonally activated and differentiated into immunoglobulin-secreting cells. This Con A-mediated helper activity was completely inhibited by the addition of alpha-methyl-D-mannoside and could not be substituted by culture supernatant of Con A-stimulated thymocytes or splenic T cells. Almost all the activity of the thymus cells was carried by peanut agglutinin low binding population. Genetic restriction between T and B cells was not observed in this helper function. In ontogeny, Con A-mediated helper activity in the thymus was first detected at a few days after birth and reached the adult level at about 1 week of age. The polyclonal helper assay system developed in the present study provides a sensitive system to analyse the helper function of thymus cells and also to delineate the early phase of the differentiation of helper T cell population.

Monoclonal antibodies (MAbs) against the optic tectum of Xenopus tadpoles were generated and screened by the immunofluorescent staining of frozen sections of tadpole brains. MAb-A5 stains the 8th and 9th plexiform layers of the optic tectum, whereas MAb-B2 stains all but the eighth and ninth plexiform layers of the optic tectum. MAb-A5 antigen is also detectable in the nucleus of Belonci, the corpus geniculatum thalamicum, the pretectal area, and the basal optic nucleus, all targets of the optic nerve, but is not detectable in the optic nerve or the optic tract. On the other hand, MAb-B2 does not stain any of these visual centers, though many fibers surrounding them are stained. Eye-enucleation experiments showed that MAb-A5 antigen is expressed in the optic tectum even when it is not innervated by optic nerves. Staining of viable brains with these MAbs indicates that these antigens are cell surface molecules. Immunoadsorption followed by SDS-PAGE suggests that proteins are constituents of these antigens. The MAb-A5 antigen in the diencephalon and the mesencephalon is not detectable at stage 35/36, but is detectable at stage 39 when the optic nerves begin to innervate the optic tectum. The spatial as well as the temporal patterns of the expression of the MAb-A5 antigen suggest that this molecule may be involved in the target recognition of optic nerve fibers.

Immunological tolerance in the mouse thymus cell population induced by the intravenous injection of deaggregated bovine gamma globulin was terminated by whole body irradiation. After irradiation, the weight of the thymus recovered biphasically, and the termination of tolerance occurred as early as in the first phase. Both Thy-1 antigen expression and helper activity of the thymus cell population in irradiated mice recovered in parallel with the recovery of the thymus weight. Sensitivity of the regenerating thymus cell to the tolerogen was not different from that of the normal thymus cell. The first phase of thymus regeneration may be caused by the proliferation and differentiation of relatively radioresistant and tolerogen insensitive precursors residing in the thymus. Tolerogen and/or immunogen reactive thymus cells may originate from the precursor.

Functions of T cells derived from single progenitor cells were investigated. B10. Thy-1.1 recipient mice were either whole body-irradiated and marrow reconstituted or thymus-shielded, irradiated and marrow reconstituted, and limited numbers (3 x 10(3) or 6 x 10(3] of a 1:1 mixture of bone marrow cells from C57BL/6 and B6.Lyt-2.1 mice were transferred intrathymically (i.t.). Donor-type (Thy-1.2+) cells of the thymus of a small portion of recipients which expressed the phenotype of either Ly-2.2 or Ly-2.1 but not both were regarded to be a clone of T cells derived from a single progenitor cell, and such clones were assayed for polyclonal helper (Th) and polyclonal cytolytic (CTL) activities as well as alloantigen-specific proliferative (mixed lymphocyte reaction; MLR) and CTL activities. Clones taken 4 weeks after transfer (4-week-old clones) which were generated in the thymus of whole body-irradiated recipients showed polyclonal CTL but not polyclonal Th activity, whereas 4-week-old clones generated in the thymus of thymus-shielded recipients showed both polyclonal CTL and Th activities. Similarly, 4-week-old clones generated in whole body-irradiated recipients responded with CTL to alloantigens when induced in the presence of T cell growth factors but not with MLR, whereas 4-week-old clones generated in thymus-shielded recipients showed both MLR and CTL to alloantigens. Repertoire diversification of 4-week-old clones, however, was incomplete, since clones generated in whole body-irradiated recipient did not necessarily respond with CTL to all alloantigens examined, and those generated in thymus-shielded recipients did not necessarily respond with MLR to all the antigens. On the other hand, T cell clones were shown to fully mature by 7 weeks after transfer in terms of cell function as well as repertoire diversification.

In the course of study to obtain murine dendritic cell lines using oncogenic retroviruses, we have established several immortalized cell lines with characteristics different from those of dendritic cells. The transformants were mainly round nonadherent cells, capable of growing in soft agar, and negative for nonspecific esterase activity. Profiles of cell surface antigens were examined by indirect immunofluorescence technique. The cell lines were positive for Fc receptor (2.4G2), J11d (J11d.2), and B220 (RA3-3A1/6.1) antigens and negative (or dull positive in small percentages) for Ia (M5/144.15.2), IL-2 receptor (3C7), Thy-1 (B5-5), Mac-1 (M1/70.-15.11.5), and macrophage (F4/80) antigens. They were negative for both surface and cytoplasmic immunoglobulins. Several clones were established from these transformant cell lines and cell surface antigens were examined. Antigenic profiles of these clones were very similar to those of the parental cell lines. Some of these clones, however, seemed to increase their Ia antigen expression. The results suggest that the transformants originated from early B-lineage cells.

Neomycin/bovine serum albumin/gold was used as a probe to detect the binding sites of aminoglycosides on the thin sections of the cochlea embedded in Spurr. The binding sites were mainly located on the stereocilia, the cuticular plate of hair cells, the head plates of Deiters' cells, fibrous structures in pillar cells, in the spiral limbus and tectorial membrane and basilar membrane, plasma membranes, mitochondria and the chromatin of various kinds of cells. Triphosphoinositide, acidic glycosaminoglycans, and RNA were considered to be responsible for the binding activity.

The development of cytotoxic T cells and helper T cells in an intact or irradiated thymus was investigated. C57BL/6 (H-2b, Thy-1.2) mice were whole body-irradiated, or were irradiated with shielding over either the thymus or right leg and tail, and were transferred with 1.5 X 10(7) bone marrow cells from B10.Thy-1.1 mice (H-2b, Thy-1.1). At various days after reconstitution, thymus cells from the recipient mice were harvested and a peanut agglutinin low-binding population was isolated. This population was further treated with anti-Thy-1.2 plus complement to remove host-derived cells and was assayed for the frequency of cytotoxic T cell precursors (CTLp) and for the activity of helper T cells (Th). In the thymus of thymus-shielded and irradiated mice, Th activity reached normal control level by day 25, whereas CTLp frequency remained at a very low level during these days. In the thymus of whole body-irradiated mice, generation of CTLp was highly accelerated while that of Th was retarded, the period required for reconstitution being 25 days and more than 42 days for CTLp and Th, respectively. Preferential development of CTLp was also seen in right leg- and tail-shielded (L-T-shielded) and irradiated recipients. Histological observation indicated that Ia+ nonlymphoid cells were well preserved in the thymus of thymus-shielded and irradiated recipients, whereas in L-T-shielded and irradiated recipients, such cells in the medulla were markedly reduced in number. These results suggest strongly that the generation of Th but not CTLp is dependent on radiosensitive thymic component(s), and that such components may represent Ia+ cells themselves in the medulla or some microenvironment related to Ia+ cells.

This study reports the establishment of cloned hybridomas between mouse myeloma cells (P3-X63-Ag8, 6.5.3) and spleen cells from BALB/c mice immunized with lung homogenates of nude mice (BALB/c nu/nu) heavily infected with Pneumocystis carinii. A hybridoma subclone, designated 1E12-8, produced antibodies of an IgG1 subclass. Using indirect immunofluorescent antibody techniques, the monoclonal antibody was found to be directed toward the pellicle antigen of air-dried P. carinii both in trophozoite and cyst forms, and to recognize the P. carinii antigen not only from nude mice but also from rats. The antibody did not cross-react with other components of the infected host lung tissue and showed little cross-reactivity with fungi examined. The monoclonal antibody should be useful in the isolation and identification of corresponding antigenic substances of P. carinii and may provide a useful tool in epidemiology, taxonomy, and diagnosis of this organism.

E-643, a newly developed alpha 1-adrenergic blocker with a postsynaptic action, was tried on 17 subjects, 10 patients with neurogenic bladder and 7 patients with lower urethral obstruction. The results of treatment were excellent in 5, good in 7, and poor in 5 of them, the rate of effectiveness therefore being 70.6%. The inhibitory effect of E-643 on the contractility of the detrusor was no stronger than that of other alpha-blockers. The rate of effectiveness for neurogenic bladders was as high as 80% whereas that for lower urethral obstruction was much lower: 57%. Concerning benign prostatic hyperplasia (B.P.H.) alone, this agent was markedly beneficial. In the case of B.P.H the urethra pressure in the prostatic urethra tends to drop and urination is facilitated even without a change in maximum urethral closure pressure, which reduces the amount of residual urine, this sequence of events taking place on condition that the contractility of the detrusor is adequate. According to urodynamic criteria, the underlying causes of the so-called bladder neck contracture can be classified into organic and functional obstruction, but it is mistaken to use this term for functional disturbance of the bladder neck. It seems worthwhile to determine the clinical effect of alpha-blockers on functional urethral obstruction. E-643 produced few if any appreciable signs or symptoms of side effects either subjectively or objectively.

Using cloned lines with the morphology of large granular lymphocytes (LGL) from BALB/c mice, we studied the exact requirements for proliferation and their functional characteristics, as well as their regulation. Although these cloned LGL lines were interleukin 2 (IL-2) dependent for growth, experiments using human recombinant IL-2 (rIL-2), known to be active on murine cells, indicated that IL-2 was a necessary but not sufficient factor. Coexistance of normal macrophages in addition to rIL-2 was found to support continuous proliferation of cloned LGL in vitro. This role of macrophages could be replaced by partially purified IL-1 derived from macrophage-conditioned medium. An IL-2 binding assay using 125I-rIL-2 suggested that the role of normal macrophages was to selectively induce and/or maintain high affinity IL-2 receptors (IL-2R) (Kd, 0.2-0.5 nM) without affecting low affinity ones (Kd, 10-30 nM). Functional studies indicated that most of the LGL clones killed various combinations of representative groups of natural killer (NK)-susceptible target cells, including leukemic cells (YAC-1, RL male 1), virus-infected cells (HeLa-measles, HeLa-herpes simplex virus), and normal bone marrow cells (BMC), whereas none of them affected any of NK-resistant target cells, including uninfected HeLa cells. Some of these clones also suppressed in vitro hematopoiesis. Such characteristic cytotoxic spectra, as well as serological phenotypes (Thy-1+, Lyt-1-2-, asialo GM1-positive, T200+, TdT-, Fc receptor-positive) indicated that these LGL clones exactly represent endogenous NK cells, rather than a variety of anomalous killer cells generated in various culture conditions. Although there was significant heterogeneity of cytotoxic spectrum among LGL clones, no clonotypic distribution of specificities was observed. Normal macrophages were found to modulate the functional expression of LGL clones. They augmented the cytotoxic potential of the clones against leukemic and virus-infected targets, but suppressed intrinsic reactivity against normal BMC. Similarly, LGL clones maintained with macrophages showed much less suppressive effect on in vitro hematopoiesis. The present observations on the interaction of cloned LGL and normal macrophages provide a basic explanation for the mechanisms by which the immediate responsiveness to IL-2 of the NK effector system, without exogenous stimulation, and the functional selectivity toward abnormal rather than normal cells, are actively maintained in vivo.

Immune complex (IC) was made of bovine serum albumin (BSA) and anti-BSA guinea pig serum, and instilled into the tubotympanic cavity of untreated guinea pigs. The location of IgG was determined by using the protein A-gold technique to trace IC in the tubotympanic mucosa of the otitis media with effusion (OME) caused by the instillation. IgG was found on the effusion, degenerated ciliated cells and on granules of goblet cells. There was no evidence of intense accumulation of IgG on the basement membrane of the mucosal epithelium or the capillaries. It is likely that IC activates complements in the tubotympanic cavity to cause OME.

We compared various strains of Propionibacterium with regard to protection of young adult mice against lethal infection with herpes simplex virus type 2 (HSV-2). Propionibacterium acnes, P. granulosum, and P. avidum were protective, while P. acidi-propionici and P. lymphophilum were ineffective. The protective effect proved to be in the cell wall fraction. Attempts were made to elucidate possible mechanisms of the protection using both effective and ineffective strains. The results strongly suggest that induction of interferon rather than activation of macrophages and natural killer cells by Propionibacterium pretreatment plays a crucial role, directly or indirectly, in protection against infection by herpes simplex virus. Propionibacterium only moderately protected newborn mice against HSV-2 infection.

The plaque-forming antibody response of C57BL/6 mice to rabbit gamma globulin (RGG) decreases as a function of ae. RGG in tolerogenic form induces tolerance of young mice but sensitizes older animals. If antigen is administered together with lipopolysaccharide, the age-dependent decline in immune responsiveness is not observed, nor are older animals sensitized by tolerogen. The age-dependent decline in immune responsiveness is due to a loss of T helper capacity; sensitization by tolerogen is attributable to a subpopulation of B cells which becomes sensitized by the tolerogen. Older animals, treated with tolerogen, show a degree of central tolerance of T cells and a relatively slight, age-dependent diminution in colchicine- or cyclophosphamide-sensitive precursors of suppressor cells.

The primary site for the growth and differentiation of B cell precursors in irradiated recipient mice was investigated. Bone marrow (BM) cells from lipopolysaccharide (LPS)-responder C57BL/6 mice were transferred into irradiated LPS-nonresponder C57BL/10ScCr mice, and the generation of donor-derived B cells in the recipient was monitored by determining the immunoglobulin-producing cells developed in the LPS-stimulated cultures of recipient's spleen cells as well as BM cells. As previously stated, the transfer of fresh BM cells resulted in the development of LPS-reactive cells both in spleen and BM simultaneously. On the other hand, when long-term cultured BM cells which were shown to be devoid of B cells and pre-B cells were used as the donor cells, the development of LPS-reactive cells was first observed only in BM, and subsequently in both BM and spleen. The failure to detect LPS-reactive cells in the spleen in the early phase, or day 11 after irradiation and reconstitution, was not attributed to the culture condition or the existence of suppressive activity in day 11 spleen cells. These results indicate that B cell precursors lodge only in the BM of irradiated recipients, grow and differentiate in the same place, and then the differentiated progeny migrate to peripheral lymphoid organs.

We report the details of our treatment of 5 patients with recessive dystrophic epidermolysis bullosa (RDEB). Good results were obtained by esophageal dilatation using the balloon catheter method. Prior to the operation, we corrected the patients' nutritional status with oral supplementation of Hinex-R as well as intravenous drip infusion of Intra-lipid, based on the nutrition index obtained by body weight, serum albumin, rapid turnover proteins, and blood minimum essence. After sufficient anesthesia of the nasopharyngeal mucosa with xylocain spray, we inserted a Microvasive Rigiflex Balloon Dilator into the stenotic region while scanning the upper GI. Then we dilated gradually by barium injection. The patients' condition improved after this treatment and follow up X-ray examination revealed no recurrence of stenosis. There were several advantages to this method: 1) It was safer and could be done repeatedly. 2) It caused less stress for patients. 3) Its effectiveness appeared rapidly. We analyzed the frequency and severity of esophageal stenosis of 12 cases with EB in our department. Ten out of 12 cases complained of subjective esophageal symptoms. Esophageal stenosis was observed in all 8 cases examined by esophagogram. We concluded that this surgical procedure for esophageal stenosis was as important as digit reconstruction in the management of RDEB.

Ribavirin and Rifamycin SV, given from birth to adult life, arrest the loss of suppressor capacity, normally occurring in older SJL mice. The two drugs differ in their effect on Th-2-type help for the indirect plaque-forming response, i.e. for help which is independent of the carrier-hapten bridge. This type of help normally increases with age. Ribavirin treatment inhibits development of Th-2-type of help; Rifamycin SV does not interfere with the development of this type of help. Neither of the two drugs had an effect on the ability of SJL mice, not treated with aggregate-freed RGG, to make a plaque-forming response to aggregated RGG.

The characteristics of natural killer (NK) activity of human peripheral blood lymphocytes treated with human leukocyte interferon (HuIFN-alpha) were compared with those of the untreated original NK activity. HuIFN-alpha, at a concentration of 50 IU/ml or more, augmented the NK activity against BALL-1, Daudi, and Namalwa target cells. On the other hand, the augmentation was less evident for the activity against Raji and Molt-4 cells. Original NK activity was detected mainly in the fraction that passed through a nylon wool column. Nylon-retained cells, however, could be activated by the addition of HuIFN-alpha to express activity against BALL-1. The activity of the nylon-passed fraction was augmented by HuIFN-alpha as well as that of unfractionated cells. NK cells could be activated by HuIFN-alpha under calcium-free conditions that allow NK cells to bind but not to lyse the target cells; augmented cytotoxicity was observed when calcium was added 2 hr later. This suggests that the augmentation of NK activity by HuIFN-alpha may have resulted from an increase in the number of NK cells.

Adult normal mice and those mice which had been thymectomized, X-irradiated, and reconstituted either with bone marrow cells and thymus cells (TxXBT) or with bone marrow cells along (TxXB), were given varying amounts (0.1-5 mg) of deaggregated soluble bovine serum albumin (sBSA). They were challenged 10 days later with an immunogenic form of BSA. TxXB mice were supplemented with normal thymus cells 3 days before the challenge. With any dose of sBSA, TxXBT and normal mice were made tolerant. Only 5 mg of sBSA, the highest dose in these experiments, was effective in inducing the tolerance in TxXB mice. The simultaneous administration of 5 mg of sBSA and 0.1 mg of alumprecipitated BSA plus 0.01 mg of endotoxin resulted in the priming in TxXBT mice but induced tolerance in TxXB mice. These results indicate that: (a) B-cell tolerance could be induced independently of the influence of T cells; (b) the tolerogen susceptibility of B cells may be lower than that of T cells; (c) such a weakly immunogenic agent as the mixture of tolerogen and immunogen could either activate the antibody response in the presence of T cells or induce B-cell tolerance in the absence of T cells.

Dopamine-beta-hydroxylase (DBH) immunohistochemistry was used to demonstrate the noradrenergic fibers and terminals in the anterior column of the rat lumbosacral spinal segments. PAP-positive varicose fibers were widely distributed in the gray matter with preferential accumulation in the nuclear regions containing motoneurons involved in the contraction of perineal striated muscles. Unmyelinated DBH fibers were composed of nodular enlargements (varicosities, 0.4-3.0 microns in diameter) and very fine, short intervals (intervaricose segments, 0.1-0.2 micron in diameter and 1.0-4.0 microns in length). DBH-positive dense products were electron microscopically often confined within small granular particles and less frequently within large granules. Additionally, in order to characterize the innervation pattern of noradrenergic fibers on dendritic bundles organized in the motoneuronal pools innervating the pelvic small muscles, semi-quantitative analysis was done in the area of the dorsolateral nucleus endowed with especially well-developed dendritic bundles. DBH terminals contacting with unreactive dendrites were more common (67.9%) than those with neuronal somata (15.1%), and the remainder (17%) had no contacts with surrounding neuronal elements. Furthermore, specialized synaptic formations were observed in only 20.1% of these nodules. The results suggest that bulbospinal descending noradrenergic neuron systems influence the functioning of pelvic muscles principally via the neuronal contacts with dendritic bundles in the spinal cord.

Neuroblastoma (NB) arises from primitive sympathetic neuroblasts in the adrenal gland or the sympathetic ganglion. NB in situ, sometimes observed in the adrenal glands of autopsied infants, is considered to be a premalignant lesion that may develop into NB. Little is understood about the morphological and biochemical changes that accompany this malignant progression. In this study, a unique monoclonal antibody, KP-NAC8, raised against a human NB cell line is described. This binds to NB cells but not to fetal neuroblasts. The antibody recognizes a Mr 200,000 surface protein on NB cells. KP-NAC8 binds to 15 of 17 human NB cell lines and all 26 fresh NB samples either from tumor tissues or from marrow aspirates involved with tumor. The antibody was found to cross-react with some other tumor cell lines, namely, Ewing's sarcoma (1 of 2), melanoma (1 of 4), lung cancer (3 of 3), and leukemia (2 of 14) cell lines. However, KP-NAC8 did not bind to any rhabdomyosarcoma (0 of 4), Wilms' tumor (0 of 4), retinoblastoma (0 of 2), glioma (0 of 4), and gastric cancer (0 of 2) cell lines examined. Among fetal tissues, KP-NAC8 did not react with normal neuroblasts in the adrenal glands of 5 fetuses. In a further study, the membrane phenotype of fetal adrenal neuroblasts was analyzed by a panel of 12 monoclonal antibodies including KP-NAC8. A comparison of the binding of the same panel of antibodies to fresh NB revealed that antibodies UJ13A, UJ127:11, PI153/3, anti-Thy-1, A2B5, BA-1, BA-2, HSAN1.2, and Leu-7 bound to both fetal adrenal neuroblasts and NB cells. Monoclonal antibodies OKIa-1 and J5 did not bind to either tissues. The only antibody that could distinguish fetal adrenal neuroblasts from NB cells was KP-NAC8. KP-NAC8 may, therefore, define a differentiation-related antigen that may prove helpful in understanding the biological nature of NB and NB in situ.

In the period of intestinal adaptation following massive resection of intestine, patients have several difficulties and problems. This study was done in 17 survivals of 31 infants and children received massive intestinal resection (residual intestine; less than 75 cm) between 1966-1987 to define these problems and difficulties. 1) Postoperative hepatic dysfunction occurred in 11 of the 17 survivals. The relationship between relapsing frequency of hepatic dysfunction and length of remaining ileal segment suggested that the reduced ileal length may be one of the etiologic factors of the hepatic dysfunction. 2) Evaluation of serum bile acid levels suggested that the disturbance of bile acid reabsorption and its rapid turn over might still remain even after catch-up growth of these patients. 3) Abnormal levels of serum minerals (Ca, InP, Mg, Zn and Cu) were observed in 4 cases during TPN, and these abnormalities disappeared by discontinuation of parenteral nutrition and beginning of oral feeding. 4) Serum vitamin levels (Vit.A, Vit.E, 25-OH-D3, and Vit.B12) still remained in abnormal even after the patients reached school age. These results suggested that very long-term nutritional cares, especially in lipid and vitamin nutrition, are necessary for the children with massive intestinal resection.