The localization of the primary visual centers in the hen diencephalon was determined by anterograde transport horseradish peroxidase (HRP) techniques. Twelve fowls (Gallus gallus domesticus) were used for HRP study and four were used for cytoarchitectural study (Nissl and Klüver-Barrera stained preparation). One-hundred microliter of 30% HRP solution in physiological saline was injected into the vitreous body of one eye of each hen under anesthesia of sodium pentobarbital (30 mg/kg body wt). After a postoperative period of 48 hours, the animals were deeply anesthetized and perfused with an injection of 1,000 ml of Ringer solution which was followed by 2,000 ml of 1% paraformaldehyde and 1.25% glutaraldehyde in a 0.1 M phosphate buffer (pH 7.4) which was then followed by 1,000 ml of 10% sucrose in the same buffer. The brain was cut into serial transverse sections of 60 microns on freezing microtome. Every section was treated with tetramethylbenzidine (TMB). Retinal projections were found in the hypothalamic area, lateral geniculate nucleus (GL), lateral part of dorsolateral thalamus (DLL), medial part of dorsolateral thalamus (DLM), ventrolateral thalamus (VLT), rostrolateral part of dorsolateral anterior thalamus (DLAlr), magnocellular part of dorsolateral anterior thalamus (DLAmc), lateral anterior thalamus (LA), ectomammillary nucleus (EM), external nucleus (NE), and nucleus superficial synencephalica (SS), contralaterally. No labeled terminals were found in the ipsilateral brain stem. In the hypothalamic region, terminals were found to be just lateral to the rostral part of the third ventriculus and the bottom of the lateral margins of the hypothalamus, which we termed medial (MRH) and lateral (LRH) retinorecipient hypothalamic nucleus. LRH had high density terminals compared with MRH and caudal MRH continued into rostral LRH so that there was no boundary between MRH and LRH in HRP preparation. MRH is contained large cells (25-35 microns in diameter) and occupied rostral 1/4 of retinorecipient of hypothalamus (RH), whereas LRH contained small type cells (about 15 microns in diameter) and occupied caudal 3/4 of RH. In the retinorecipient nuclei of the thalamus, high density terminals were found in GL, LA, DLAlr, NE, SS and EM. In DLAlr and EM, granulae of HRP product were bigger than in other terminal nuclei and also the density of those terminals was high. GL and LA have large nuclei which receive retinal afferents. Labeled terminals of those nuclei were distributed homogeneously throughout the whole nucleus except for the inner layer of GL. Cytoarchitectonically, GL was divided into two layers.

The expressions of several cytokeratins (CKs) in the outer root sheath (ORS) of the human anagen hair follicles were immunohistochemically studied using 11 antikeratin monoclonal antibodies (MoAbs) and 10 specimens from scalp. CKs 1, 10, 11, which are markers for differentiating keratinocytes, were exclusively found in the intermediate cells and the granular cells at the infundibulum. In cytokeratin expression, a distinct linear demarcation between the infundibulum and the isthmus was observed. Trichilemmal keratinization appeared to go in an inner-upward direction toward the hair canal. CK 19, a marker of undifferentiated stem cells, was found in outermost cells of the ORS at the isthmus and in some cells of the lower ORS. CK 16, a marker of hyperproliferative keratin, was detected in the outermost cells of the infundibulum and all the cells of the ORS below the isthmus. Therefore, the keratinocytes at the infundibulum may show a differentiation similar to that of the interfollicular epidermal keratinocytes. The ORS cells below the isthmus seem to move up to inner-upward direction along the hair axis with differentiation.

To investigate whether each of cisplatin (CDDP) and carboplatin (CBDCA) was AUC (Area Under the Curve)-dependent or time-dependent drug, Human Tumor Clonogenic Assays (HTCA) were performed in various exposure times of CDDP or CBDCA using PC-9 cells. From the result of this study, it was shown that CDDP and CBDCA were AUC dependent drugs. Furthermore, to evaluate the combination effect of CDDP and CBDCA, we carried out HTCA using PC-9 and PC-14 (human lung adenocarcinoma cell lines) cells, and evaluated the combination effect by the median effect analysis of Chou, T.C. and Talalay, P. The combination effects were above the additive effects at the AUC ratios of free Pt in the exposure medium of 3.2, 6.5, 13.1 (CBDCA/CDDP). Especially in the combination of 3 hours exposure of CBDCA followed by 1 hour exposure of CDDP, the Combination Index by median effect analysis was from 0.74 to 0.86 at the AUC ratio of 13.1. But the combination effect was antagonistic at that of 19.5. CDDP and CBDCA, which are AUC dependent drugs, have shown different side effects in previous clinical practice. And the combination effect of both drugs is more than additive effect at the AUC ratios of free Pt of 3.2, 6.5, 13.1 (CBDCA/CDDP). It is considered that the combination chemotherapy of CDDP and CBDCA for patients of lung adenocarcinoma may be useful.

An adult T cell leukemia associated with pure red cell aplasia-like lesion was described in this paper. A 51 year-old woman was admitted because of headache and palpitation in October 1988. On admission, physical examination showed marked pallor but no detectable superficial lymphadenopathies. Hepatosplenomegaly was not observed. The blood examination revealed normocytic anemia with Hb of 6.6 g/dl and marked leukocytosis of 18,800/microliters with 43% ATL cells. The bone marrow aspirate showed moderate infiltration of ATL cells and a few erythroblasts. The bone marrow biopsy disclosed moderate infiltration of ATL cells, only a few erythroblasts with maturation arrest and marked fibrosis. The erythropoietin in serum was elevated (686 IU/microliters). To clarify the mechanism of development of the PRCA-like lesion, the peripheral blood lymphocytes (ATL cells) or serum of the patient was added to in vitro erythroid colony formation. The patient's serum increased BFU-E but either serum or lymphocytes didn't inhibit the growth of CFU-E compared with control.

It is well known that Epidermal Growth Factor (EGF) is a cell-regulating factor for variety of tissues in vitro including normal and malignant cells. Furthermore, Takano et al reported that a decreased expression of EGF receptor in clones of human cancer KB cell line might be one of the pleiotropic properties of multidrug-resistant cells. However, both the influence of EGF on human urological cancer cell lines and the relation between EGF receptors and sensitivities of antitumor drugs on these cell lines have not been fully described. We have studied the effects of EGF on growth of 4 transitional carcinoma cell lines of bladder (TCCaB), 1 squamous cell carcinoma cell line of bladder (SCCaB), 5 renal cell carcinoma cell lines (RCC) and 3 prostatic carcinoma cell lines (CaP), as well as the relationship between the number of EGF receptors and drug sensitivities of these cell lines in vitro against methotrexate, vinblastine, adriamycin, cisplatin and etoposide (VP16). The present results determined by the in vitro colony forming efficiency method showed that exogenous addition of EGF to cell cultures at 0.1 ng/ml stimulated the growth of SCCaB by 169.0%, and at 1 ng/ml inhibited that of RCC by 2.9%-79.0%, relative to control. The more EGF receptors by 125I-EGF binding assay, the higher inhibition of VP16 on the growth of these cell lines. These results suggested that EGF stimulated the growth of SCCaB and inhibited the growth of RCC in vitro, and we found that these phenomena were correlated with neither the number of EGF receptors nor affinities of that receptors.(ABSTRACT TRUNCATED AT 250 WORDS)

Both in vitro and in vivo effects of lithium on mouse hematopoiesis were investigated. The addition of 1 mmol lithium carbonate to the culture enhanced in vitro colony formation of the granulocyte macrophage precursor (CFU-GM) by 26%, the megakaryocyte precursor (CFU-Meg) by 29%, and the erythrocyte precursor (CFU-E) by 46% as compared to the control culture without lithium carbonate. Lithium resulted in an increase of colony size for both the CFU-Meg and the CFU-E. Oral administration of lithium chloride at daily doses of 0.3 mg caused a significant increase in granulocytes 3 to 12 days after the start of administration. It also resulted in a mild elevation of the platelet count, but it had no influence on hematocrit. The number of the CFU-GM increased transiently on day 3 of the lithium administration and subsequently returned to the pretreatment level. Furthermore lithium accelerated the recovery of the granulocyte count and bone marrow CFU-GM content after 300 rad total body X ray irradiation.

The stem cell number is one of the important factor determining tumor control radiation dose. It is now clear that 50% tumor cell doses in various rodent tumors inversely related to their tumor control radiation doses. Roles of radiation-sterilized cells on the clonogenicity of survived stem cells are controversial, and should be explored by future research. Methodological innovations are also desirable to measure the stem cell number in human tumors.

A patient with pure red cell aplasia (PRCA), who had the inhibitor to erythroid precursors in serum, is described. A 72-year-old female was referred to Nagoya National Hospital because of progressing anemia in April 1988. On admission, her hemoglobin was 4.8 g/dl, reticulocyte 0.8%, and bone marrow specimen contained only 1.2% erythroblast. On these bases, she was diagnosed as pure red cell aplasia. After small amount of blood was transfused, her hemoglobin and erythroblast in bone marrow (EBM) increased to 7.8 g/dl and 39.1%, respectively, and she was discharged. However, after a month, her hemoglobin dropped to 4.6 g/dl, reticulocyte to 0.1%, and EBM to 0%. Soon after corticosteroid therapy (prednisolone, 40 mg, daily) was started, a marked elevation of reticulocyte count was observed, and then her hemoglobin increased to 11.0 g/dl, and EBM to 31.6%. The reason for a transient spontaneous remission at the onset of her disease was occurred is unclear. The number of BFU-E in her bone marrow was within normal range, but it was suppressed significantly (65%) after the addition of her serum and the complement purified from rabbit plasma. This finding suggest the presence of inhibitor to erythroid precursors in her serum.

Bone marrow and spleen are the major hematopoietic tissue in adult mice. However, little is known about the specific mechanism regulating hematopoiesis within these tissues. Since Dexter et al. first described conditions to maintain bone marrow hematopoiesis, long term bone marrow culture (LTBMC) has been developed in order to analyze the mechanism of the maintenance of proliferation and differentiation of hematopoietic stem cells in vitro. Furthermore, several stromal cell lines which are able to support the growth and differentiation of hematopoietic lineage, has been established from LTBMC. Although it is well known that bone marrow stromal cell lines are able to produce colony stimulating factors, it has been suggested that the stromal cell factors which involve membrane bound moieties must have a key role in the regulation of hematopoiesis. We expect that monoclonal antibodies to the surface of bone marrow stromal cells could detect such a critical stroma-associated protein that bounds the cell surface of the bone marrow stroma.

The levels of peripheral progenitor cells was measured serially after cancer chemotherapy in 4 patients with non-Hodgkin's lymphoma and one patient with rhabdomyosarcoma who received recombinant human granulocyte colony-stimulating factor (rG-CSF). This study was composed of two independent phases: in the first phase, patients received a course of cytotoxic chemotherapy only, and in the second phase, they received the same chemotherapy followed by subcutaneous injection of rG-CSF (2 micrograms/kg/day) for 10-14 days. In the control phase, the levels of granulocyte-macrophage colony-forming units (CFU-GM) and erythroid burst-forming units (BFU-E) per milliliter increased during the early recovery phase, but rG-CSF treatment increased the number of CFU-GM 3 to 18-folds, and the number of BFU-E increased 1.3 to 4.6-folds. An overshoot in the blood progenitor levels occurred at the day 8-10 of rG-CSF administration. And then, the peak of neutrophil count followed 3-5 days later. After the discontinuation of rG-CSF, the number of blood CFU-GM and BFU-E fell rapidly. This results suggest that in vivo expansion of circulating hemopoietic progenitors can be achieved by the administration of rG-CSF, and this approach might be clinically applicable to cancer patients who are a candidate of peripheral blood stem cell autotransplantation.

Granulocyte colony-stimulating factor is a 18 kD glycoprotein hormone which exhibits the following biological actions; (1) amplification and release into blood of bone marrow hematopoietic stem/progenitor cells, (2) enhancement of neutrophil production and stimulation of their release into blood and into tissues, (3) prolongation of neutrophil survival and activation of their various functions, (4) stimulation of myeloid leukemia cell growth. In patients with malignant neoplastic disorders, therefore, its recombinant human form (rhG-CSF) may be an effective strategy for various therapeutic purposes. These include reduction of the risk for infectious complications, more effective killing of malignant cells, and promotion of bone marrow transplantation. We have recently performed several clinical studies of rhG-CSF derived from Chinese hamster ovary cells. Results so far obtained from these studies strongly indicate that rhG-CSF can be safely and efficiently used for the above purposes.

High-dose chemotherapy with autologous hematopoietic stem cell transplantation (HD-CT with AHSCT) has been studying to overcome drug resistance and to obtain higher antitumor efficacy on hematological malignancies and solid tumors. The annual rate of transplants is increasing steadily. This method has boon now performed safely owing to the development of researches about hematopoietic growth factors and stem cells. It was demonstrated that the administration of recombinant human granulocyte colony stimulating factor (rhG-CSF) and granulocyte Macrophage colony stimulating factor (rhGM-CSF) could shortened the duration of leukopenia. Autologous peripheral stem cell transplantation has the advantage for the earlier bone marrow reconstitution and the unnecessary of purging. Recently, it was found that rhG-CSF and rhGM-CSF could increase the number of peripheral blood stem cells, so further studies are expected. For recurrent breast cancers resistant to conventional chemotherapy, response rates of 60-80% may be achieved using HD-CT with AHSCT, but the duration of response is generally short. So, trials consisted of induction conventional chemotherapy followed by HD-CT with AHSCT in the earlier course of the illness have conducted to obtain higher complete response (CR) rate and survival rate. Although the observed duration is short, higher CR rate has been achieved in several clinical studies. There are still remaining with many problems to be settled, HD-CT with AHSCT may have the potentiality to cure not only recurrent breast cancer but also another malignancies.

We described clinical and neuropathological findings of a case of primary cerebral lymphoma with central neurogenic hyperventilation (CNH). A 54-year-old awake woman with a primary cerebral lymphoma presented hyperventilation for two weeks. Arterial blood gas showed severe respiratory alkalosis; PH 7.603, PaCO2, 10.5 mmHg, PaO2 129.8 mmHg, HCO2 10.4 mmol/L, BE -8.0, O2SAT 98.9%. Rebreathing from a paper bag, and intravenous administration of diazepam and sodium bicarbonate failed to alter the respiratory pattern. Consecutive CAT scans indicated that CNH didn't occur when the tumor extensively invaded the cerebral cortex, cerebellum, thalamus and basal ganglia but was initiated when the lymphoma invaded the brain stem. Pathological study showed lymphoma cells invaded the cerebral cortex, cerebellum, thalamus and basal ganglia severely, and the brain stem moderately and recently. Contrary to the cases reported by Plum, Lange and Bateman, the lower medulla was also involved. Possible mechanisms for CNH are discussed in relation to the pathological findings and consecutive CAT scan findings.

A case of cerebellar ganglioglioma in a 5 year-old girl is presented. She came to our hospital on January 30, 1989 with complaints of headache of one year duration. CT scans disclosed a low density lesion suggesting a cystic tumor in the left cerebellar hemisphere with moderate hydrocephalus. Preoperative MRI demonstrated more clearly the location and extent of the tumor. She was operated on using suboccipital craniotomy, on March 3. Subtotal removal of the tumor was performed because the tumor had invaded the brain stem. She made an uneventful recovery without any neurological deficits. Histologically, the tumor was composed of ganglion cells and astrocytic cells, so it was diagnosed as ganglioglioma. Cerebellar ganglioglioma is a rare tumor, and only 17 cases have been reported including the present case. Clinical and radiological study of these cases revealed that there are no specific findings to indicate cerebellar ganglioglioma and preoperative diagnosis is impossible. But practically, MRI is the most sensitive method for identifying the extent of the lesion and, thus, is of benefit for deciding operative strategy.

A case of herpes simplex encephalitis with high signal intensity area in brainstem on MRI was presented. A 44 year-old woman suffered from oral aphthous ulcerations in the end of 1988, and then it improved naturally. Oral aphtha appeared again on February 1988 followed by resistant fever to antibiotics and right hemiparesis. She was admitted to our hospital on 25 February 1988. Neurological examination revealed mild consciousness disturbance, neck stiffness, right-side deviation of tongue with dysarthria and right hemiparesis with bilateral plantar extensor reflex. Right hemisensory deficit in all modalities and truncal ataxia was also detected. Some aphthous ulcerations were revealed in oral cavity, but there were no ulcers on genitalia nor uveitis. CSF showed 32 mononuclear cells/mm3, protein 52 mg/dl and glucose 97 mg/dl. CSF culture and india ink stain, and serum autoantibodies were all negative. EEG and CT scan with contrast enhancement showed no significant abnormalities. T2-weighted brain MRI revealed high intensity area in the center of the pons. Anti-herpes simplex virus (HSV) type I antibody titer (FA method) in both serum and CSF were highly positive. Neurological symptoms gradually improved on the therapy of aciclovir and adrenal cortico-steroid. High intensity area in the pons on MRI was also gradually reduced. In this case, complete diagnostic differentiation from neuro-Behçet disease was difficult, but this case did not meet its diagnostic criteria. From the change of anti-HSV antibody titer both in serum and CSF, we diagnosed this case HSV brainstem encephalitis presenting high intensity area in the pons on MRI which has never been reported.