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Purpose The objective of this study was to elucidate the impact on clinical outcomes resulting from re-irradiation for locally recurrent (LR) brain metastases (BM) using CyberKnife® stereotactic radiosurgery (SRS). Materials and methods Seventy-seven patients with 254 LR BM lesions treated using SRS re-irradiation between January 2014 and December 2018 were analysed in this retrospective study. The local control (LC), overall survival (OS) rates, and adverse events were assessed. The adverse events were classified according to the Common terminology for adverse event (CTCAE) v5.0. Results The median follow-up duration was 8.9 months. The median age of the patients was 55 years (IQR: 47–62). The 3, 6, and 9-month LC and OS rates were 92.2%, 73.4%, and 73.4% and 79.2%, 61.0%, and 48.1%, respectively. On multivariate analysis the gender (male vs. female; HR, 1.79; 95% CI, 1.06–3.01; P = 0.028), type of first brain radiation (WBI vs. SRS) followed by re-irradiation using SRS (HR, 9.32; 95% CI, 2.77–15.27; P 12cc vs. ≤ 12cc; HR, 1.84; 95% CI, 1.10–3.11; P = 0.02), and recursive partitioning analysis (RPA) (I vs. II & III; HR, 0.38; 95% CI, 0.19–0.70; P = 0.001) were independent predictive factor for OS. Radionecrosis was reported in 3 patients. Conclusion With acceptable toxicity, SRS re-irradiation for LR BM showed a favourable rate for LC and OS and reported better OS for the female gender, a patient undergoing first brain radiation with SRS, tumour volume ≤ 12cc, and RPA-I. This result needs to be further evaluated in future clinical studies.

Acid-sensing ion channel 4 (ASIC4) belongs to the ASIC gene family of neuronal proton-gated cation channels, and is the least understood subtype among the members. Previous studies of ASIC4 expression in the mammalian central nervous system have shown that ASIC4 is abundantly expressed in the spinal cord and in various brain regions, such as the cerebral cortex, the hippocampus, and the cerebellum. However, the detailed distribution of ASIC4 transcripts in mammalian brains still remains to be elucidated. In the present study, radioactive in situ hybridization histochemistry with an ASIC4-specific cRNA probe was performed on wild-type mouse brains, followed by X-gal staining experiments with Asic4-lacZ reporter mice Asic4tm1a(KOMP)Mbp. It was found that ASIC4 mRNAs were widely expressed throughout the wild-type brain, but preferentially concentrated in the olfactory bulb, the piriform cortex, the caudate putamen, the preoptic area, the paraventricular nucleus, the medial habenular nucleus, the pretectal area, the lateral geniculate nucleus, the amygdaloid complex, the superior colliculus, the interpeduncular nucleus, and the granule cell layer of the ventral hippocampus, and these results were in agreement with the X-gal-positive reactions observed in the mutant brain. In addition, X-gal staining combined with immunohistochemistry identified intense signals for ASIC4 transcriptional activity in most of the choline acetyltransferase (ChAT)-positive principal neurons located in the basal forebrain cholinergic nuclei. Our data provide useful information to speculate possible roles of ASIC4 in diverse brain functions.

We report a case of successful treatment of a ruptured distal aortic arch aneurysm with cardiac tamponade by using selective cerebral perfusion for protecting the brain. A 79-year-old man had sudden onset of severe chest and back pain. Chest computed tomography (CT) suggested an acute aortic dissection. He was immediately transferred to the emergency room of our hospital. Echocardiography performed on admission revealed intrapericardial fluid, and hemodynamic monitoring suggested cardiac tamponade. After pericardiocentesis and removal of 400 ml bloody fluid, his hemodynamic condition became stable. Enhanced chest CT showed ruptured distal aortic arch aneurysm with pericardial and pleural effusion. Emergency patch plasty of the aneurysm under extracorporeal circulation (ECC) was performed, assisted by selective cerebral perfusion and deep hypothermia. The patient's postoperative course was uneventful, except for minor transient respiratory troubles, and he was able return to his usual activity.

Male PD strain rats are sterile in the homozygous condition (pd/pd) due to abnormal spermatogenesis detectable at around nine weeks of age. Previous studies have indicated electron microscopic abnormalities in the Sertoli cells of pd/pd males at three and 12 weeks of age. Since spermatogenesis and Sertoli cell function depend on gonadotropins (luteinizing and follicle-stimulating hormones [LH and FSH]) and testosterone, production and/or secretion of these hormones might be altered in pd/pd males. The aim of the study reported here was to investigate the hormonal status of pd/pd males at three, six, and nine weeks of age. Although alteration was not evident in the LH-immunoreactive cells, FSH-immunoreactive cells in pd/pd males were small in size with scant cytoplasm and were reduced in number and area (73 and 51% of phenotypically normal pd/+ males, respectively) at three weeks of age, although serum FSH concentration was similar to that in pd/+ males. At six and nine weeks of age, percentages of the areas occupied by LH- and FSH-immunoreactive cells in pd/pd males were higher than those in pd/+ males. Serum FSH concentration in pd/pd males was significantly high at nine weeks of age, although a difference in serum LH and testosterone concentration was not evident. These results suggest that FSH production in pd/pd males is decreased at three weeks of age. This might be associated with the Sertoli cell abnormalities and subsequent abnormal spermatogenesis seen in adult life.

Background: Most Japanese pediatric neurologists attempt other treatments before using adrenocorticotropic hormone (ACTH) therapy for West syndrome (WS), and even then, they use only a low-dose synthetic ACTH to avoid serious adverse effects. In this multi-institutional study, the authors analyzed the initial effects, adverse effects, and long-term outcome in patients treated with low-dose synthetic ACTH in Japan. Methods: The medical records of 138 patients with WS, who were treated with low-dose synthetic ACTH therapy for the first time at the authors’ institutions between 1989 and 1998, were analyzed. Results: At the end of ACTH therapy, excellent effect on seizures was noted in 106 of 138 (76%) patients, good effect in 23 (17%), and poor effect in 9 (7%). Initial effects on EEG were excellent in 53 of 138 (38%) patients, good in 76 (55%), and poor in 9 (7%). As for seizure prognosis at the time of follow-up, 51 of 99 (52%) patients were seizure-free, whereas 48 (48%) patients had seizures. Mental outcome was normal in 6 of 98 (6%) patients, mild mental retardation in 16 (16%), moderate mental retardation in 26 (27%), and severe mental retardation in 50 (51%). The initial effects of ACTH on seizures and long-term outcome were not dose dependent (daily dosage 0.005 to 0.032 mg/kg, 0.2 to 1.28 IU/kg; total dosage 0.1 to 0.87 mg/kg, 4 to 34.8 IU/kg). The severity of adverse effects correlated with total dosage of ACTH, and the severity of brain volume loss due to ACTH correlated well with the daily dosage and total dosage of ACTH. Conclusion: Low-dose synthetic ACTH therapy is as effective for the treatment of WS as the higher doses used in previous studies. The dosage of synthetic ACTH used in the treatment of WS can be decreased as much as possible to avoid serious adverse effects.

Significance of Wnt signaling with beta-catenin mutations on solid-pseudopapillary neoplasm (SPN) of the pancreas was studied by immunohistochemistry and molecular analysis. On immunohistochemistry, all 18 SPNs tested showed diffuse cytoplasmic/nuclear positivity for beta-catenin. Upon direct DNA sequencing of exon 3 of the beta-catenin gene, 15 (83%) of the 18 SPNs showed 1-bp missense mutation in codons 32 (5 cases), 33 (3 cases), 34 (3 cases), 37 (3 cases), and 41 (1 case). Immunoreactivity for cyclin D1, one of the intranuclear targets of beta-catenin complexes, was found in tumor cells of more than half the tumor cells of all of the 18 SPNs. The present study strongly suggested a significant role of Wnt signaling, mostly associated with beta-catenin mutations in the tumorigenesis of SPN.

BACKGROUND—Human β-defensin 2 (hBD-2) plays a role in the innate defence system at mucosal surfaces. Colonisation of Helicobacter pylori in the stomach is an important pathological factor in gastrointestinal illnesses, including gastritis, peptic ulcer, and gastric adenocarcinoma. AIMS—To evaluate the antibacterial role of hBD-2 against H pylori infection in the gastric mucosa. SUBJECTS—Biopsied gastric mucosa specimens from H pylori positive (n=6) and H pylori negative (n=6) individuals were used. H pylori was determined by the presence of urease activity and microscopic examination. METHODS—The specimens were examined for hBD-2 expression by reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemistry, and in situ hybridisation. The antibacterial effect of hBD-2 against H pylori was evaluated by the number of colony forming units of H pylori after incubation with 0, 10−9, 10−8, 10−7, 10−6, or 10−5 M of hBD-2 peptide. RESULTS—All six H pylori positive specimens expressed a high level of hBD-2 mRNA while hBD-2 mRNA was not detected in the H pylori negative specimens by RT-PCR. Immunohistochemistry using anti-hBD-2 antiserum revealed that hBD-2 was expressed in the surface epithelium of H pylori infected specimens. In gastric specimens obtained after H pylori eradication, hBD-2 immunoreactivity had dramatically decreased. In situ hybridisation confirmed that hBD-2 transcripts were localised in the epithelium of H pylori infected gastric specimens. Incubation with hBD-2 reduced the growth rate of cultured H pylori in a dose dependent manner, and incubation with 10−5 M hBD-2 completely inhibited the proliferation of H pylori. CONCLUSIONS—H pylori infection induces hBD-2 expression in the human gastric epithelium. hBD-2 inhibited the growth of H pylori in vitro, suggesting that hBD-2 plays an antibacterial role in H pylori induced gastritis. Keywords: human β-defensin 2; Helicobacter pylori; gastritis; antimicrobial peptide

Myoclonus can be caused from a variety of central nervous system disorders. With regard to its pathophysiology, much work has been done in adult patients, but reports from pediatric clinics are scarce. We describe the clinical and electrophysiological features of four different types of myoclonus in children using back averaging technique. In a patient with cortical reflex myoclonus, myoclonic jerks were induced in the distal right arm by taps on the ipsilateral fingers. Myoclonus was preceded by the cortical spike which was localized posterior to the central sulcus contralateral to the myoclonus, and the cortical spike was time-locked to the myoclonus. In a patient with subcortico-cortical myoclonus, myoclonus occurred as a focal motor seizure. Myoclonic jerks were preceded by generalized cortical waves which were of maximal amplitude over the cortex contralateral to the myoclonus, and cortical waves time-locked to the myoclonus. A patient with spinal myoclonus had rhythmical and segmental myoclonus. Myoclonus was induced in both arms after electrical stimulation at the wrist with a latency which was slightly longer than that of the F wave. In a patient with brainstem myoclonus, muscle activity was bilaterally synchronous and was predominant in the proximal flexors. The pattern of muscle recruitment suggested that the myoclonus signal travelled downward along the spinal cord and upward through the brainstem. The back averaging technique is essential in the differential diagnosis of myoclonus.

Our previous studies using 12-week-old PD (pd/pd) male rats have indicated high incidence of morphologically abnormal Sertoli cells. Because sterility in these males is attributable to abnormal spermatogenesis, it is suggested that the abnormal Sertoli cells are related to abnormal spermatogenesis. In this study, the testes of prepubertal 7- and 21-day-old male pd/pd rats were examined ultrastructurally for the presence of abnormal Sertoli cells. The pd/+ males served as controls. At 7 days of age, light and dark types of Sertoli cells were found in pd/pd males. However, no anatomic abnormality existed in those cells. Cytoplasmic organelles were underdeveloped. The only difference between the light and dark Sertoli cells was the high electron density in the nucleus and cytoplasm of the dark cells. These features were similar between pd/pd and pd/+ males. At 21 days of age, the dark Sertoli cells had disappeared, and light Sertoli cells and abnormal Sertoli cells were observed in pd/pd males. The light Sertoli cells had well-developed cytoplasmic organelles. On the other hand, abnormal Sertoli cells had a dark appearance, irregularly shaped nuclei, and a large quantity of lipid droplets in the cytoplasm. These findings suggested the presence of functional disorders in lipid metabolism. The light and abnormal Sertoli cells also were found in pd/+ males; however, incidence of the abnormal Sertoli cells was significantly higher in pd/pd males (11.3%), compared with that in pd/+ males (4.9%). Thus, these results indicate that in pd/pd male rats, a number of abnormal Sertoli cells appear by 21 days of age.

A patient with post-hypoxic brainstem reflex myoclonus was examined. In spontaneous jerks electromyographic activity started in the sternocleidomastoid and spread down the spinal cord bilaterally. A similar jerk was elicited by somaesthetic or acoustic stimulation. The patient's blink reflex suggested bilateral pontine lesions at least. Thus, the pattern of muscle activation pointed to a brainstem origin of the myoclonus. Spinal conduction studies conducted during the jerks suggested that the myoclonus signal facilitated a path identical to the spinobulbospinal reflex demonstrated in animals.

Anti-intercellular adhesion molecule-1 (anti-ICAM-1) and anti-lymphocyte-function-associated antigen-1 (anti-LFA-1) monoclonal antibodies (mAbs) were injected into mice and their effects on tumor metastasis were investigated using two murine models. Depending on the dose, the anti-ICAM-1 mAb (KAT-1) expressed both inhibitory and promoting effects on liver metastases of ICAM-1 + LFA-1 + P815 mastocytoma cells, whereas it enhanced lung metastases of the ICAM-1-LFA-1-Meth-A fibrosarcoma cells at any doses. In contrast, anti-LFA-1 mAb (KBA) showed promoting effects only on the metastases of both tumor lines. Treatment of mice with either mAb enhanced metastases of P815 mastocytoma cells in the spleen.

Pseudosarcomatous myofibroblastic tumor (PMT) is the result of reactive proliferation of myofibroblasts. In children, PMT of the urinary bladder can be mistaken for embryonal rhabdomyosarcoma clinically, radiologically, and by light microscopy. We are reporting the clinical, histological, and immunohistological features of 11 patients with childhood PMT of urinary bladder that were diagnosed initially as a sarcoma, usually rhabdomyosarcoma. The morphologic spectrum of PMT is broad, with mixtures of myxoid, leiomyomatous, and sclerosing matrix patterns, the myxoid type being the most common. The proliferating cells consist of three forms of myofibroblastic cells: long spindle cells (type I), intermediate spindle cells (type II), and ganglion-like cells (type III), together with various types of inflammatory cells. The immunohistologic profile of the proliferating cells was characterized by positive reactions to vimentin, muscle-specific actin, alpha-smooth-muscle actin, polyclonal desmin, and keratin. Ultrastructural studies showed myofibroblastic differentiation of the tumor cells. No patients have had metastases or local recurrence. Histologic, immunohistochemical, and clinical data from 71 cases of PMT, including the 11 cases in this report, confirm the benign behavior of these lesions. The etiology of these lesions is unclear, including the absence of surgical or other trauma in all of the children.

We reported clinicopathological, histopathological and immunohistological features of two suggestive cases with post transfusion graft-versus-host disease (GVHD) and examined. In addition, we have declared histopathologic characteristics of acute and chronic GVHD with literatural consideration. On the basis of a review of two cases, we came to the following conclusion; It is implied that early lymphoid proliferation may be useful to make a diagnosis of acute GVHD. Case 2 had pathological confirmation of acute GVHD in the liver and small intestine. In the liver, a damage of interlobular bile ducts and T lymphocytes infiltration close to ducts were the most specific feature of acute GVHD. From an immunohistological study in case 2, it was confirmed that abnormal expression of HLA-DR antigen and the appearance of activated T lymphocytes played an important role in the development of GVHD reaction in the liver.

The postoperative survival of patients with preoperatively identified stage III N2 non-small cell cancer (NSCLC) is poor. A pilot study of induction chemotherapy consisting of cisplatin, mitomycin C and vindesine (CMV) was undertaken in an attempt to improve the curative potential of surgery in this group of patients.Thirty-two patients with stage III N2a NSCLC received two cycles of CMV, (80 mg/m2 cisplatin i.v. on day 1, 8 mg/m2 mitomycin C i.v. on day 1, and 3 mg/m2 vindesine i.v. on day 1 and 8, every 4 weeks) preoperatively. The responding patients received a further two courses of CMV after surgery.The response rate was 63% (20 PR). All patients underwent operation including radical mediastinal node dissection. Twenty-six resections were curative and six were absolute non-curative. The toxicity was generally mild, but there were two hospital deaths, due to bronchopleural fistula and postoperative pneumonia. Sixteen patients have shown recurrent disease up to now. When the recurrence patterns were classified by clinical response, 11 responding patients were disease-free (9-84 months), but all non-responding patients had recurrent diseases. The median survival of curatively resected patients with pathological N2 was 39 months with a 5-year survival of 29%.This induction chemotherapy was well tolerated and revealed a high response rate. The current survival is obviously improved compared with the historical control, indicating that subsequent phase III clinical trials are urgently needed.

A 36-year-old man was diagnosed as having acute non-lymphocytic leukemia (AML M5b) in 1985 and received allogenic bone marrow transplantation from an ABO-mismatched sibling in January 1987. Recovery of erythropoiesis in this patient was delayed, then the hemoglobin level improved in parallel with disappearance of anti-A antibody in the serum on day 260 post transplantation. However, as anemia occurred again despite no relapse of leukemia on day 350, we tried to determine the presence of erythropoietic inhibitory factor in this patients. Erythroid colony formation was decreased when bone marrow cells were co-cultured with peripheral mononuclear cells from the patient. Further, erythroid colony inhibitory activity was found in conditioned medium of PHA-stimulated T cells from the patient. Sephadex gel fractionation showed that the molecular weight of this inhibitory factor was approximately 11,000 and addition of a high concentration of EPO did not eliminate the inhibitory activity. These findings suggest that the novel inhibitor described in this manuscript, produced by T cells, was differed from previously reported inhibitors such as anti-EPO antibody, spermine and uremic toxins.

Effect of tumor necrosis factor-alpha (TNF), which is supposed to be one of the mediators responsible for endotoxin, on metallothionein (MT) synthesis was determined in various tissues of rats. At 18 hr after the injection of TNF, MT-I concentrations significantly increased in the liver, lung and heart, but not in the kidney and thymus. In the spleen of TNF-treated rats, MT-I concentrations decreased. Thus, synthesis of MT-I was tissue specific and dose-dependent. In the liver, the synthesis of both metallothionein-I and metallothionein-II was observed and the rate of the induction was greater in the latter. Antioxidative role of the MT synthesized by TNF was discussed.

Acute lymphoblastic leukemia (ALL) patients with a Philadelphia chromosome (Ph+ ALL) were treated with a combination of antineoplastic drugs recommended for both myeloid and lymphoid leukemia (BHAC-DMPV: behenoylcytosine arabinoside, daunorubicin, 6-mercaptopurine, prednisolone, and vincristine). Ph+ ALL patients with chromosome breaks which occur within the major breakpoint cluster region (M-BCR rearranged Ph+ ALL) were treated with natural interferon-alpha (IFN-alpha) after entering complete remission. In this study, four of seven patients with Ph+ ALL had M-BCR rearrangement, and all achieved complete remission with karyotypic normalization. Subsequent cytogenetic analysis during complete remission in two ALL patients with M-BCR rearrangement revealed that the percentage of bone marrow cells with the Ph chromosome increased, while the bone marrow maintained remission status. This cytogenetic-hematological discrepancy led us to consider that M-BCR rearranged Ph+ ALL might be a variant of chronic myelogenous leukemia, therefore, three Ph+ ALL patients with M-BCR rearrangement were treated with IFN-alpha after achieving complete remission. In contrast, only one of three patients with M-BCR non-rearranged Ph+ ALL obtained complete remission.

In the PD strain of rats, male pd homozygotes never sire, whereas male heterozygotes and female hetero- and homozygotes are fertile. In order to investigate the cause of infertility, pd/pd males were examined for their reproductive ability and gross and histological changes in the reproductive organs. Mating ability of pd/pd males was comparable to that of pd/+ males when a 2 week cohabitation procedure was employed. Cryptorchidism, which is considered to be a cause of infertility, was observed in 63% of the 12 week old pd/pd males. However, this abnormality frequently was observed unilaterally and the incidence of bilateral cryptorchidism was only 13%. Although the relative weights of descended testes and epididymides were comparable between the pd/pd and pd/+ males at 3 and 6 weeks of age, the values at 9 and 12 weeks of age were significantly lower in pd/pd males than in pd/+ males. A marked difference was not noted in relative weight changes of the seminal vesicles and prostate between the pd/pd and pd/+ males. Histological examination revealed defects in spermatogenesis in both descended and undescended testes in pd/pd males. The changes first appeared at 9 weeks of age and included vacuolation of nuclei of spermatocytes and spermatids, degeneration of spermatids, and occasionally multinucleated giant cells in the seminiferous tubules. In accessory sexual organs such as the epididymides, seminal vesicles, and prostate, no histological abnormalities were detected. These findings indicate that the substantial cause of infertility in pd/pd males is a disorder of spermatogenesis through which functionally normal spermatozoa are not produced.

Cytogenetic analyses were performed on 12 adult patients with abnormal megakaryoblastic proliferation which was detected by ultrastructural cytochemical study (platelet peroxidase) and platelet-megakaryocytes-specific monoclonal antibodies (TP-80, Plt1, AN51, and KOR-77). The patients consisted of two patients with myelodysplastic syndromes (MDS), three with acute megakaryoblastic leukemia (AMKL), six with megakaryoblastic transformation in Philadelphia-positive chronic myelogenous leukemia (CML-meg-BC), and one case of chronic myeloproliferative disorder (CMPD). Among them, an inversion of the long arm of chromosome 3 [inv(3)(q21q26)] was found in one AMKL patient with a normal platelet count. Chromosome change at band 3q26 was also found in one MDS patient without thrombocythemia. Furthermore, the long arm of chromosome 13, where rearrangements in myelofibrosis are clustered (13q12----q22) was seen in one MDS patient. Trisomoy of chromosome 19 was found in one AMKL patient and three CML-meg-BC patients. These findings indicate that cytogenetic abnormalities involving 3q26, 13q, and trisomy 19 are associated with hematologic neoplasia with megakaryocytic lineage in adult patients, although these abnormalities were not related to the survival of the patients. During the period of this study, two acute myelogenous leukemia patients (AML-M2 and AML-M5b) with chromosome rearrangements at band 3q21 and thrombocythemia were found, indicating that chromosome abnormality at band 3q21 is related to quantitative platelet dysfunction, whereas that at 3q26 is related to hematologic malignancies with a proliferation of megakaryocytic lineage.

Five children with renovascular hypertension were treated with percutaneous transluminal angioplasty, and followed for 12 to 32 months. Blood pressure fell to normal in four patients and improved in one. Angiographic and endocrinological improvements were associated with a fall in blood pressure and no complications were observed. Angioplasty is considered a safe, effective treatment for children with renovascular hypertension.

Five monoclonal antibodies to human (h) FSH have been prepared, isolated, and characterized. They were produced by hybridomas derived from FO myeloma cells and spleen cells from mice immunized with hFSH or its beta-subunit (hFSH beta). Two of the antibodies (A101 and A102) which recognized the alpha-subunit of hFSH bound much better when alpha was associated with the beta-subunit forming the intact hFSH molecule than when alpha was in free form. These antibodies showed 9-10%, 2-3%, and 1-3% cross-reactions with alpha-subunits in hTSH, hCG, and hLH, respectively. Two antibodies (B201 and B202) recognized only free beta-subunit. One antibody (B305) recognized free beta-subunit and native hFSH. There was no significant cross-reaction of these antibodies to FSH beta with hTSH, hLH, and hCG. Using solid phase competitive binding and sandwich assays, we compared the epitopes for these antibodies. Antibodies A101 and A102 recognize the same epitope on hFSH alpha. Antibodies B201 and B202 recognize different epitopes, but they seemed to be adjacent. Antibody B305 bound a different epitope than B201 and B202. Such characteristics of these antibodies can be useful for sensitive and specific assay of hFSH or hFSH beta and also may be helpful in studying FSH interaction with its receptor.

Sinomenine, an epimorphinan alkaloid, was tested for the immunosuppressive effect in mice. This compound produced a decrease of plaque-forming cells (PFC) to a T cell-dependent antigen, sheep red blood cells, in vivo. The depression of the PFC response induced with sinomenine was dose and time dependent. On the other hand, it failed to suppress the PFC response to a T cell-independent antigen, lipopolysaccharide. The immunosuppressive dose of sinomenine did not alter the cellularity of spleen, thymus, bone marrow and peripheral blood leucocytes, the DNA synthesis activity of bone marrow cells nor the proliferative responses of spleen cells induced by T cell and B cell mitogens in unprimed mice. These data suggest a selective effect of sinomenine on lymphoid cells. This compound has a potential for use in studies of immuno-deficiencies or clarifing some aspect of immunity.

The presence of tumor-specific or tumor-associated antigens in primary urinary bladder tumors induced by N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) in ACI/N rats was examined by means of mixed lymphocyte-tumor cell culture. Bladder tumors (papilloma with cancer foci or cancer) induced by BBN treatment for 18 weeks or more (BT), epithelial cells from papillary hyperplasia induced by 6-week BBN treatment (PH), and normal bladder epithelial cells (NBE) were isolated from the bladder and used as stimulator cells after mitomycin C treatment. BT, PH and NBE all strongly stimulated the blastogenesis of the autologous lymphocytes on culture in medium containing fetal calf serum (FCS), but the spleen cells also stimulated the autologous lymphocytes in this medium. Although the magnitudes of blastogenesis were lower than those in FCS medium, on culture in medium containing rat serum, BT, PH and NBE, but not spleen cells, stimulated the blastogenesis of the autologous lymphocytes. The response with BT was significantly higher than the response with NBE, but there was no statistically significant difference between the response with BT and the response with PH. These results suggest that FCS medium, but not medium with rat serum, evokes the blastogenic response of lymphocytes cultured with autologous spleen cells, and that blastogenic responses with autologous bladder cancer can be produced by stimulation with some sort of tissue antigen present in normal or pre-neoplastic urinary bladder epithelial cells.

Characteristic storage inclusions of Gaucher cells contain numerous tubular structures and are demonstrated enzyme-cytochemically to be transformed secondary lysosomes. Tubular structures are composed mainly of twisted multilayers and partially of flat layers with or without twisting. Glucocerebroside extracted and purified biochemically from the Gaucher spleen resembles these flat layers. Therefore, the flat layers are considered to be a fundamental fine structural unit of the tubular structures made by crystallization of glucocerebroside molecules. The tubular structures are observed by electron microscopy to be formed in the digestive process of blood cells, especially erythrocytes, in the lysosomes of Gaucher cells. In one culture experiment, monocytes obtained from Gaucher patients developed tubular structures in the phagolysosomes after ingesting heat-denatured erythrocytes and became Gaucher cells. In the other culture experiment, monocytes were also transformed into Gaucher cells by taking up tubular structures added to the culture medium. Thus, both the intralysosomal formation of tubular structures in the macrophages and their ingestion of extracellular tubular structures released from other Gaucher cells seem essential for the development of Gaucher cells.

The characteristics of the cytotoxic cells induced by ip injections of an immunoadjuvant, OK-432 (Picibanil), into ACI/N rats bearing syngeneic bladder cancer, BC47, were examined. The cytostatic activity, but not the cytolytic activity, of peritoneal macrophages was augmented when either normal or cancer-bearing rats were treated with OK-432. In contrast, the plastic nonadherent cells of the peritoneal exudate cells from OK-432-treated cancer bearing rats, but not lymph node cells or spleen cells, killed all ACI/N rat bladder cancers tested as well as ACI/N rat hepatoma cells and Meth-A mouse sarcoma cells. The plastic nonadherent cells from OK-432-treated normal rats also killed hepatoma cells and Meth-A cells, but not bladder cancer cells. The cytolytic cells that were induced in cancer-bearing rats by OK-432 treatment and showed cytolytic activity specific for bladder cancer were found to be sensitive to anti-rat thymocyte serum and complement, nylon-adherent, and Fc receptor-negative. The cells that showed nonselective cytolytic activity were nylon-adherent and insensitive to anti-rat thymocyte serum and complement.

A rare case of clear cell carcinoma of the palatal salivary gland in a 69-year-old man is reported. Histopathologically, nests and cords of tumor cells, which showed predominantly clear cytoplasms, and occasionally eosinophilic cytoplasms, proliferated on the overlying epithelium and capsules of the fibrous connective tissue. The clear cell cytoplasms were found to be PAS positive, diastase soluble, and Alcian-blue negative, indicating the presence of glycogen. Ultrastructurally, the tumor cells were found to contain glycogen and cytoplasmic filaments in various proportions. These observations suggest that clear cell carcinoma is myoepithelial in origin.

The effect of multilamellar vesicles (MLV) on reticuloendothelial (RE) activity, as measured by the carbon clearance method, was investigated in mice. MLV were prepared with a mixture of dipalmitoyl phosphatidylcholine, cholesterol and a charged agent (7:2:1 molar ratio). RE activity was moderately depressed shortly (1-4 hr) after i.v. injection of all MLV tested; negatively charged MLV with dicetyl phosphate, positively charged MLV with stearylamine and neutral MLV without a charged agent. In the later time, either MLV with dicetyl phosphate or MLV with stearylamine caused a marked stimulation of reticulo-endothelial system (RES), but neutral MLV did not. A peak of RE activity resulted from the treatment of MLV with dicetyl phosphate appeared 48 hr after injection and then gradually returned to the normal level within 7 days. The depression and the enhancement of RE activity induced by the treatment of MLV with dicetyl phosphate were observed to be dose-related and significant at a dose of 1 mg/mouse and 0.3 mg/mouse, respectively. These results should be taken into consideration upon the use of liposomes as a drug carrier, or for blocking RES.

In vitro cytotoxicity against tumor cells of lymphocytes in sc implanted BC47 bladder tumor of ACI/N rats with or without Propionibacterium avidum (P. avidum) treatment was studied. Tumor-associated lymphoid cells (TAL) were obtained from tumor tissues by mechanical treatment (NDi fraction) and by enzymatic treatment with Dispase I, a proteolytic enzyme (Di fraction), followed by passage through glass wool columns to deplete tumor cells. NDi fraction of TAL from P. avidum-treated animals showed a significant cytolytic activity against BC47 cells, but not against other ACI/N bladder tumor cell lines, BC12 and BC50. These TAL lost the cytolytic activity on treatment with anti-rat thymocyte serum or anti-rat T cell monoclonal antibodies, R1-3B3 and R1-10B5, and complement. Natural killer activity determined with YAC-1 cells was low in the cells of NDi fraction and scarcely detectable in the cells of Di fraction from both P. avidum-treated and untreated rats. These results indicate that the antigen-specific cytotoxic T cells in the tumor in situ are induced by in vivo P. avidum treatment. On the other hand, P. avidum treatment augmented nonspecific cytolytic activity of peripheral lymphoid cells such as plastic-nonadherent peritoneal cells, spleen cells and blood lymphocytes in normal and BC47-bearing rats. However, the antigen-specific cytolytic T cells were predominantly induced and recovered in the plastic nonadherent peritoneal cells of BC47-bearing rats by the treatment with P. avidum.

The clinical efficacy and the safety of fluconazole as given at an intravenous dose of 100-400 mg daily were assessed in 7 patients with deep mycosis associated with hematological malignancy. Enrolled in the study were 1 patient with acute lymphatic leukemia, 1 with acute myelocytic leukemia, 2 with acute myelomonocytic leukemia, 2 with malignant lymphoma and 1 with myelodysplastic syndrome. Pathogens isolated from 4 patients were all Candida species including 2 Candida albicans, 1 Candida parapsilosis and 1 Candida krusei. Diagnoses of fungal infections of the patients were Candida pneumonia in 3 patients, candidemia in 1 and fungemia suggested in 3. Assessment of the clinical efficacy was made on 4 patients from whom pathogens were isolated. The global clinical improvement was good in 2 patients and fair in 1 with Candida pneumonia and good in 1 with candidemia. In the mycological assessment, pathogenic fungi were eradicated in 3 patients and decreased in 1 patient. No significant adverse reactions nor abnormality in clinical laboratory tests related with the dosing of fluconazole were observed in any of the patients.

Sonicated liposomes containing actinomycin D in the membranes were chemically coated with the subunits of monoclonal immunoglobulin M (IgM) antibody against a mouse mammary tumor-associated antigen (MM antigen) and examined for their in vitro and in vivo antitumor effects against MM46 (MM+) and MM48 (MM-) tumors of C3H/He mouse origin. The antibody-bearing, actinomycin D-containing liposomes (chemoimmunoliposomes) were selectively bound to MM+ tumor cells and showed much more in vitro cytotoxicity against the tumor cells than that shown by free actinomycin D. The in vivo antitumor effect of the chemoimmunoliposomes was tested on the mammary tumor cells (5 X 10(4) to 5 X 10(6) transplanted i.p. into syngeneic mice. A single i.p. injection of the chemoimmunoliposomes containing 0.3, 0.5, or 1 microgram of actinomycin D into MM46 tumor-bearing mice resulted in the cure of some mice and a prolonged survival time in the rest of the mice as compared to results in controls. In this test, free actinomycin D, anti-MM IgM antibody, and bovine serum albumin-coated liposomes containing actinomycin D were marginally effective or ineffective. To examine a systemic antitumor effect of chemoimmunoliposomes, mice were inoculated with MM46 tumor cells and then treated with a single i.v. injection of liposomes 4 days later. If the mice were pretreated with an i.v. injection of unmodified multilamellar liposomes, an injection of the chemoimmunoliposomes containing 1 microgram of actinomycin D resulted in a significant inhibition of tumor growth. Both free actinomycin D and bovine serum albumin-coated liposomes containing actinomycin D were ineffective against the s.c. tumor. These results indicate that an antitumor drug entrapped in the membranes of small sonicated liposomes bearing antitumor monoclonal antibodies can be delivered to antigenic tumor cells and exert more efficient antitumor activity than does the free drug.