In vivo function of immune inhibitory molecule B7-H4 in alloimmune responses.

B7 ligands deliver both costimulatory and coinhibitory signals to the CD28 family of receptors on T lymphocytes, the balance between which determines the ultimate immune response. Although B7-H4, a recently discovered member of the B7 family, is known to negatively regulate T cell immunity in autoimmunity and cancer, its role in solid organ allograft rejection and tolerance has not been established. Targeting the B7-H4 molecule by a blocking antibody or use of B7-H4(-/-) mice as recipients of fully MHC-mismatched cardiac allografts did not affect graft survival. However, B7-H4 blockade resulted in accelerated allograft rejection in CD28-deficient recipients. B7-1/B7-2-double-deficient recipients are truly independent of CD28/CTLA-4:B7 signals and usually accept MHC-mismatched heart allografts. Blockade of B7-H4 in these mice also precipitated rejection, demonstrating regulatory function of this molecule independent of an intact CD28/CTLA-4:B7 costimulatory pathway. Accelerated allograft rejection was always accompanied by increased frequencies of alloreactive IFN-γ-, IL-4- and Granzyme B-producing splenocytes. Finally, intact recipient, but not donor, B7-H4 is essential for prolongation of allograft survival by blocking CD28/CTLA4:B7 pathway using CTLA4-Ig. These data are the first to provide evidence of the regulatory effects of B7-H4 in alloimmune responses in a murine model of solid organ transplantation.

Teratoma in a feline unilateral cryptorchid testis.

A 14- x 12- x 6-cm (approximately 600 g) cryptorchid testis was surgically removed with the omentum from the posterior portion of the abdominal cavity of a 2-year-old male American Shorthair cat. At the time of a previous orchiectomy, a unilateral testis had been discovered in the scrotum. The cut surface of the cryptorchid testis consisted of solid areas with many cysts containing fluid and spongy soft areas. Histologically, the cryptorchid testis was composed of neuroectodermal components and diffuse immature glial tissues, cystic, tubular, and papillary epithelial tissues, and immature cartilaginous tissues. A metastatic mass in the omentum had histologic structures similar to those of the cryptorchid testis. The present case was diagnosed as teratoma in a feline unilateral cryptorchid testis.

Expression of GFRalpha-1, receptor for GDNF, in rat brain capillary during postnatal development of the BBB.

It is well known that the blood-brain barrier (BBB) matures at approximately 2 wk after birth in the rat. Recently, we showed that glial cell line-derived neurotrophic factor (GDNF) enhances the barrier function of porcine endothelial cells forming the BBB in culture. In the present study, we examined the relation between permeability of the BBB, using Evans blue as a tracer, and expression of the GDNF family receptor (GFRalpha-1) during postnatal development of the BBB. Morphometric analysis showed that exudation of Evans blue from capillaries of the cerebral cortex progressively decreased until postnatal day 21. Inversely, immunohistochemical examinations showed expression of GFRalpha-1 in the capillaries at postnatal day 3 and expression that reached the same levels as observed in adult rats by postnatal day 10. However, c-ret, which is thought to mediate a signal evoked by binding of GDNF to GFRalpha-1, was not expressed in the capillaries of the brain cortex in 3-mo-old rats. On the other hand, the tight junction proteins occludin and ZO-1 appeared to be fully expressed at birth. The reciprocal relation between GFRalpha-1 expression and the permeability of the BBB strongly suggests active participation of GDNF in postnatal development of the BBB, although the mechanism(s) involved is still veiled.

Effects of light on the photosynthetic apparatus and a novel type of degradation of the photosystem I peripheral antenna complexes under darkness.

A novel type of degradation of photosystem I peripheral antenna complexes has been observed in rice leaves under darkness in the present study. Photosynthesis, chlorophyll content, the chlorophyll a/b ratio, and relative amounts of ribulose-1,5-bisphosphate carboxylase/oxygenase decrease during dark treatment. The levels of photosystem II reaction-center complex and cytochrome f on the basis of units of chlorophyll also decline rapidly under darkness. In contrast, the levels of photosystem I reaction-center complex remain stable under darkness for six days. Low-temperature fluorescence emission spectra ascribed to photosystem I antennae clearly show a blue shift. A similar shift is also observed in the photosystem I complexes resolved with dodecyl maltoside-polyacrylamide gel electrophoresis. Moreover, polypeptide analysis of the thylakoids and photosystem I complexes isolated from the green gels shows that some polypeptides originating from photosystem I peripheral antenna complexes disappear during the dark treatment. A curve-fitting method also displays remarkable changes in the chlorophyll components between the light and dark treatments. It is likely that these results indicate the disconnection/disassembly of the photosystem I antenna as well as the photosystem II complexes induced by dark treatment. Moreover, these findings also imply the existence of different degradation mechanisms for the photosystem I and II complexes.

Glial cell line-derived neurotrophic factor induces barrier function of endothelial cells forming the blood-brain barrier.

Since a deep involvement of astrocytes, a kind of glial cells, in differentiation of the blood-brain barrier (BBB) has been suggested, we examined the relation of glial cell line-derived neurotrophic factor (GDNF) to the BBB. First, immunohistochemical examination of the cerebral cortex of rats revealed that glial cell line-derived neurotrophic factor receptor (GFRalpha1) was preferentially expressed on the cell membranes of capillary endothelial cells. Second, to elucidate the effects of GDNF on the BBB, capillary endothelial cells isolated from the porcine cerebral cortex were cultured and then changes in tight junction function of the endothelial cells were examined after addition of GDNF, in terms of transendothelial electrical resistance (TER) and permeability. GDNF at concentrations of 0.1 and 1 ng/ml significantly activated the barrier function of the endothelial cells in the presence of cAMP. Since GDNF is secreted from astrocytes sheathing capillary endothelial cells in the brain cortex, our results strongly suggest that GDNF enhances the barrier function of tight junctions of the BBB on the one hand, and also supports the survival of neurons on the other hand.

DNA helicase III of Saccharomyces cerevisiae, encoded by YER176w (HEL1), highly unwinds covalently closed, circular DNA in the presence of a DNA topoisomerase and yRF-A.

Previously, we have purified and characterized DNA helicase III from the yeast Saccharomyces cerevisiae [Shimizu, K. and Sugino, A. (1993) J. Biol. Chem. 268, 9578-9584]. Here, we have further characterized DNA helicase III activity. It was found that the combined action of the helicase III, yeast DNA topoisomerase I (yTop I), and yeast RPA protein on a covalently closed, circular DNA generates a highly underwound DNA species that has been called form I* or form U. Furthermore, these underwound structures can be accessed by yeast DNA polymerase I (alpha)-primase to initiate DNA synthesis. These reactions mimic in vivo initiation of chromosomal DNA replication. In order to clone the gene encoding DNA helicase III, a partial amino acid sequence of the purified DNA helicase III polypeptide was determined. Using a mix oligonucleotides synthesized based on the amino acid sequence of the helicase, we cloned the gene encoding the helicase III and found it to be identical to YER176W (HEL1) on chromosome V. The amino acid sequence predicted from the nucleotide sequence of the gene has conserved DNA helicase domains that are highly homologous to those of DNA helicases required for DNA replication. However, complete deletion of the gene from the chromosome did not result in any growth defect, suggesting that the gene product is not required for DNA synthesis or that it is functionally substituted by other helicase(s). Furthermore, the deletion strain does not exhibit sensitivity to any DNA-damaging reagents, although it is hypersensitive to calcofluor white, hygromycin, and papulacandin.

[Effects on the grieving process of the families of people who use home care stations].

The aim of this study was to clarify the effects on the grieving process of bereaved families, and to investigate what visiting nurses should do to provide support. Twelve cases in which nurses from our station provided home care in 1998 were analyzed with regard to the grieving state of the families, the families' physical conditions, social situations, and support systems. The results showed that the number of carers during the patient's period of illness, the support system, and level of communication with medical providers affected the grieving process. To minimize the load on the grieving family and help them achieve closure at an early stage, it is necessary to begin developing care ability and preparing a support system during the period of home treatment. Sufficient emotional care must also be provided for the family, including education in preparing for death. The families lean heavily on medical staff for this, and building a good relationship is therefore vital.

[Oral and dental health and oral and dental support of home patients--role of dental hygienist in the home service nursing station].

Home patients have few chances for going out, so communication with their family means a lot. Talking and eating are particular pleasures. Therefore, oral and dental health and oral and dental support are very important for home patients. A dental hygienist from our clinic visits and offers oral and dental health (oral care) and oral and dental support (oral rehabilitation) to home patients as part of a care plan with home care nurses. Moreover, as general conditions are closely related with oral function, maintaining oral and dental health and regular oral and dental support are very important in order to improve the quality of life (QOL) of home patients.

The 1.5 GHz electromagnetic near-field used for cellular phones does not promote rat liver carcinogenesis in a medium-term liver bioassay.

We have recently established that local exposure to a 929.2 MHz electromagnetic near-field, used for cellular phones, does not promote rat liver carcinogenesis in a medium-term bioassay system. In the present study, a 1.439 GHz electromagnetic near-field (EMF), another microwave band employed for cellular phones in Japan, was similarly investigated. Time division multiple access (TDMA) signals for the Personal Digital Cellular (PDC) Japanese cellular telephone standard system were directed to rats through a quarter-wavelength monopole antenna. Numerical dosimetry showed that the peak SARs within the liver were 1.91-0.937 W/kg, while the whole-body average specific absorption rates (SARs) were 0.680-0.453 W/kg, when the time-averaged antenna radiation power was 0.33 W. Exposure was for 90 min a day, 5 days a week, over 6 weeks, to male F344 rats given a single dose of diethylnitrosamine (200 mg/kg, i.p.) 2 weeks previously. At week 3, all rats were subjected to a two-thirds partial hepatectomy. At week 8, the experiment was terminated and the animals were killed. Carcinogenic potential was scored by comparing the numbers and areas of the induced glutathione S-transferase placental form (GST-P)-positive foci in the livers of exposed (48) and sham-exposed rats (48). Despite increased serum levels of corticosterone, adrenocorticotropic hormone (ACTH) and melatonin, the numbers and the areas of GST-P-positive foci were not significantly altered by the exposure. These findings clearly indicated that local body exposure to a 1.439 GHz EMF, as in the case of a 929.2 MHz field, has no promoting effect on rat liver carcinogenesis in the present model.

Lack of promoting effects of the electromagnetic near-field used for cellular phones (929.2 MHz) on rat liver carcinogenesis in a medium-term liver bioassay.

The possible cancer promotion potential of local exposure to a pulse modulated 929.2 MHz electromagnetic near-field on chemically-initiated rat liver carcinogenesis was investigated employing a medium-term bioassay. A 929.2-MHz electromagnetic near-field of time division multiple access (TDMA) signal for PDC (Personal Digital Cellular, Japanese cellular telephone standard) system was directed to rats through a quarter-wavelength monopole antenna. Maximum local specific absorption rates (SARs) on temporal average were 7.2-6.6 W/kg within the whole body and 2.0-1.7 W/kg within the liver, which was the target organ. The whole-body average SARs on temporal average were 0.80-0.58 W/kg. Temporal peak SARs had three times these values due to the duty ratio of the PDC signal. Exposure was for 90 min a day, 5 days a week, over 6 weeks. The exposure apparatus was specially designed for this experiment, to allow exposure of the lateral mid-section of the rat body to the electromagnetic near-field. Male F344 rats, 6 week-old, were initially (at week 0) given a single dose of diethylnitrosamine (DEN, 200 mg/kg body wt, i.p.). At 2 weeks later, exposure (48 rats) or sham-exposure (48 rats) was started. The exposure of electromagnetic near-fields was performed using the exposure apparatus mentioned above. At week 3, all rats were subjected to a 2/3 partial hepatectomy. At week 8 (i.e. after 6 weeks exposure or sham-exposure), the experiment was terminated and all rats were killed. Carcinogenic potential was scored by comparing the numbers and areas of the induced glutathione S-transferase placental form (GST-P) positive foci in the livers of the exposed and sham-exposed rats. A further group of 24 animals, given only DEN and partial hepatectomy, served as the controls. The numbers (no./cm2) of GST-P positive foci were 4.61 +/- 1.77, 5.21 +/- 1.92 (P < 0.05, versus control) and 4.09 +/- 1.47 and the areas (mm2/cm2) were 0.30 +/- 0.16, 0.36 +/- 0.21 and 0.28 +/- 0.15, for the exposed, sham-exposed and control groups, respectively. There were no significant differences between the exposed and sham-exposed groups. These findings clearly indicated that local body exposure to a 929.2-MHz field, modulated in a PDC waveform, has no significant effect on rat liver carcinogenesis under the experimental conditions employed.

[The meaning of eat in home patients with end-stage cancer].

When a patient difficult to eat, he will not be only malnutrition, but loses a volition of his life and falls into depression. This time, we present a patient with relapse advanced pharynx cancer, who could not ingestion. Therefore she had said "DEATH". She tried eagerly rehabilitation of ingestion, swallowing and breathing, and she was able to eat again and could be positive posture. Ingestion is very important for human, and the patient found a meaning of her life and she was able to take home care nursing.

Venous malformation in the posterior fossa: guidelines for treatment.

Venous malformations in the posterior fossa are relatively rare. Although the introduction of CT and MRI has made them easier to detect, their treatment is still controversial. Based on our experience with six patients and a review of the literature, we have tried to establish guidelines for their treatment. Since they have a benign natural course and may provide venous drainage in the posterior fossa, venous malformation found incidentally, unruptured venous malformation with nonhaemorrhagic complications, and those accompanied by small intracerebellar haematoma of less than 2 cm in diameter due to their rupture, should be treated conservatively unless they are associated with a coexistent malformation. Venous malformation with intracerebellar haematoma larger than 3 cm or reexpansion of the haematoma due to rebleeding should be treated surgically by evacuation of the haematoma. Resection of venous malformations in the posterior fossa should be restricted to cases in which the malformation is small and does not serve as a functional venous drainage route. Radiation therapy, including the "gamma-knife", may be a treatment of choice in the future.

High sensitivity of LEC rats with chronic hepatitis to hepatocarcinogenesis: decreases in unscheduled and replicative DNA synthesis of the hepatocytes.

We carried out the following three experiments to clarify the mechanism of hepatocarcinogenesis in Long-Evans Cinnamon (LEC) rats. (1) Sensitivity to diethylnitrosamine (DEN): LEC rats (8 and 25 weeks old) without and with hepatitis and age-matched F344 rats were administered an intraperitoneal injection of a low dose of DEN. Eight weeks after the injection, the numbers of glutathione-S-transferase placental-form (GST-P)-positive foci in the 33-week-old LEC rat liver were significantly higher than those in the livers of the other three groups of rats. (2) Potential for unscheduled DNA synthesis (UDS): Isolated hepatocytes of 25-week-old LEC rats with chronic hepatitis showed about one-third the level of UDS induced by UV irradiation, as compared to that of age-matched F344 rats, while no significant difference was found between the UDS of isolated hepatocytes of 8-week-old LEC rats and age-matched F344 rats. (3) Potential for proliferation: Isolated hepatocytes from 8-week-old LEC rats responded well to epidermal growth factor (EGF) in culture, to almost the same degree as F344 rat hepatocytes, while a remarkable decrease in the responsiveness of hepatocytes isolated from 25-week-old LEC rats to EGF was found. These results suggested that LEC rat hepatocellular carcinoma could be naturally initiated after the onset of hepatitis by carcinogens contaminating food and the environment, probably due to the reduction of DNA repair activity, after which initiated hepatocytes selectively proliferate in response to growth stimuli endogenously produced as a result of continuous loss of hepatocytes (chronic hepatitis), because of a decrease in growth activity of non-initiated hepatocytes.

[Predictive factors for in-hospital mortality and delayed mortality following aneurysmal subarachnoid hemorrhage].

It is important to detect predictive factors for in-hospital and delayed mortality of patients with subarachnoid hemorrhage (SAH) due to ruptured aneurysm. Forty-eight patients with initial bleeding of aneurysmal SAH were referred to our hospital from January 1982 to December 1985. In-hospital mortality was 16.7% (8 to 48), and 15% (6) of forty patients died later during the follow-up period. Using the Kaplan-Meier method we were able to conclude that, cumulatively, there was 70.8% probability that much patients would survive for 10 years. We analysed predictive factors of in-hospital and delayed mortality retrospectively. The most significant predictive factor for in-hospital mortality was SAH grading on admission, and for delayed mortality (29.2%) age on admission was the best predictive factor. In fact, two patients died with cardiac event during the follow-up period. This result suggests that, although the SAH grading on admission was the second most significant factor for delayed mortality, patients who survived in the acute phase had a survival probability similar to those in a normal control group.

[Ossifying fibroma in the cranial vault. Case report].

Ossifying fibroma is relatively common in the maxilla, but rare in the cranial vault. A 10-year-old girl was referred for painful swelling of the left temporal region. On admission, she presented no abnormal physical and neurological findings except for the painful swelling. Plain skull X-ray films showed a radiolucent lesion of the left temporal bone about 4 cm in diameter, with a hyperostotic area of the parietal side. Computed tomography scan using bone window level also showed an abnormal density lesion in the same site. Curettage of this tumor was performed from a cosmetic point of view and at the family's petition. Histological examination showed vascular fibrous tissue in which lamellar bone was surrounded by osteoblasts.

[Gonadotropin, as a tumor marker, in body fluid and tumor tissues of germ cell tumors].

The value of gonadotropin in the body fluids of germ cell tumor patients is its usefulness as a tumor marker. It is also used for differential diagnosis and/or judgement of therapeutic effects. In order to clarify the most effective value of gonadotropin as a tumor marker in the body fluids, we compared the value in serum, liquor and urine with one another. The liquor contained highest (1650 IU/l) value of gonadotropin in the primary intracranial germ cell tumors, mostly in choriocarcinoma. But the gonadotropin value was highest (3050 IU/l) in the serum of secondary intracranial choriocarcinoma. Chiasmal germ cell tumor, except choriocarcinoma, which does frequently secrete gonadotropin (alpha, beta) showed moderate or very high values in the liquor. However, pineal germ cell tumors rarely secrete gonadotropin and sometimes mild high value are obtained in the serum without gonadotropin secretion immunohistochemically. In such cases, the gonadotropin may be increased by indirect mechanism of gonadotropin-secretion following pineal disorder. In most of such cases, the gonadotropin was not human chorionic gonadotropin (HCG) but lutein hormone (LH). Because alpha-subunit of such gonadotropin has the same structure, their antibodies show immunologic cross reaction. So, a count of beta-subunit gonadotropin in the serum or liquor is the best way for differential diagnosis or judgement of therapeutic effects. From our results, it is considered that the tumor secretes HCG if the serum beta-HCG value was higher than 30 IU/l, and that it doesn't secrete HCG if beta-HCG value was lower than 10 IU/l or non calculable. The mild increased HCG may be caused by hypothalamo-diencephalic disorder such as pineal tumor.(ABSTRACT TRUNCATED AT 250 WORDS)

Large intramedullary lipoma of the cervical cord and posterior fossa. Case report.

A large intramedullary lipoma of the cervical cord extending into the posterior fossa is reported in a 7-year-old boy. Magnetic resonance imaging was very useful for delineation of the anatomy of the lipoma as an aid in planning the operation.