Effects of tyrosine administration on serum biopterin in normal controls and patients with Parkinson's disease.

After administration of tyrosine, total concentration of biopterin, the cofactor for tyrosine hydroxylase, was increased in the striatum, adrenal glands, and serum of rats, and in the serum of humans. Serum biopterin is lower in patients with Parkinson's disease than in normal controls. After oral administration of tyrosine, the increase in serum biopterin concentration was smaller in patients with Parkinson's disease (less than twofold) than in healthy controls (three-to sevenfold). These results suggest that tyrosine may have a regulatory role in biopterin biosynthesis and that patients with Parkinson's disease may have some abnormality in the regulation of biopterin biosynthesis.

Tumor-associated mucin-type glycoprotein (CA54/61) defined by two monoclonal antibodies (MA54 and MA61) in ovarian cancers.

Two monoclonal antibodies, MA54 and MA61, were established by immunizing with culture medium supernatants of a lung adenocarcinoma cell line, and a double-determinant sandwich enzyme immunoassay system was developed by using these two monoclonal antibodies. The antigen recognized by this assay (CA54/61) was found to be often high in the sera of several cancers. The antigen recognized by MA54 (CA54) or MA61 (CA61) proved to be carbohydrate chain on a high molecular weight mucin-type glycoprotein, and CA54 has NeuAc alpha 2-6galactose in the terminal residue. CA54/61 was frequently found in the sera of ovarian cancer patients, the positive rate being 67, 64, 40, and 78% in serous, mucinous, endometrioid, and mesonephroid cancers, respectively, when the cut off value was set at mean + 4 SD. Since the positive rate of CA125, which is now the most widely used for the diagnosis of ovarian cancers, is rather low (approximately less than 50%) in mucinous cystoadenocarcinoma, CA54/61 will be of clinical value. In addition, CA61 was detected immunohistochemically in the fetal red blood cells with nuclei, indicating its oncodevelopmental character in nature.

Human monoclonal antibody (HMST-1) against lacto-series type 1 chain and expression of the chain in uterine endometrial cancers.

A human monoclonal antibody termed HMST-1 was produced by fusing lymphocytes from segments of human pelvic lymph nodes from an endometrial cancer patient with murine myeloma cells. The epitope recognized by HMST-1 was determined to be lacto-series type 1 chain-containing glycosphingolipid (Gal beta 1-3GlcNAc beta 1-3Gal beta 1-4Glc beta 1-1Cer) by isolating the antigen from endometrial cancer cell line SNG-II and analyzing with fast atom bombardment mass spectrometry, permethylation analysis, and exoglycosidase treatment. By the immunohistochemical avidin-biotin-peroxidase complex method, no normal endometrium and benign endometrial hyperplasia were stained with HMST-1, but HMST-1 reacted with about 35% of endometrial cancer cases. These facts indicate that the rate of expression of the antigen increases along with the course of malignancy in the endometrium. By sialidase treatment of the section, the positive rate increased to 57% in endometrial cancers and to 13% in normal endometrium, indicating that the antigen was masked with sialic acid and exposed by neuraminidase treatment. Immunohistochemistry also revealed that the antibody reacted with human fetal alimentary tract epithelium and mesothelium, indicating the oncodevelopmental nature of Gal beta 1-3GlcNAc beta 1-3Gal beta 1-4Glc beta 1-1Cer.

Cancer-associated galactosyltransferase as a new tumor marker for ovarian clear cell carcinoma.

Serum cancer-associated galactosyltransferase antigen (caGT) was assayed in gynecological cancer patients by means of a GT-II-reactive monoclonal antibody (MAb 3872)-based immunoassay. Thirty-six of 47 (75%) ovarian cancer patients showed a significant elevation of caGT in serum above the cutoff level of 200 milliunits/ml (mean +/- 2 SD) determined from normal controls. Particularly, serum caGT levels in eight of nine patients with ovarian clear cell carcinoma were above the cutoff value, and six of them gave more than 200 milliunits/ml. Elevation of caGT in serum from pregnant women was also detected, and the level increased during the course of gestation. Immunohistochemical study revealed that not only various ovarian carcinoma cells in vivo and in vitro, but also syncytiotrophoblast of early gestational placenta, fetal tissues such as mucus-producing cells in the lower alimentary tract, and renal tubules at the 11th week of gestation were stained with MAb 3872, thus indicating its oncofetal character. Compared with CA-125, caGT showed a lower false-positive rate (10%) in benign gynecological diseases, and there was no correlation between caGT and CA-125 values. Therefore, caGT will be a useful tumor marker for ovarian cancers, especially for clear cell carcinoma.

Monocytic differentiation modulates apoptotic response to cytotoxic anti-Fas antibody and tumor necrosis factor alpha in human monoblast U937 cells.

Interferon-gamma (IFN-gamma), vitamin D3 (VD), and retinoic acid (RA) induce differentiation of human monoblastic leukemia U937 cells to macrophage-like cells with potential superoxide anion-generating activity upon further stimulation. Here we report that U937 cells thus differentiated show various responses to apoptotic induction with a cytotoxic anti-Fas antibody and tumor necrosis factor (TNF). VD-or RA-treated U937 cells acquired resistance against Fas- or TNF receptor (TNFR)-mediated apoptosis, whereas apoptotic cell death was accelerated in IFN-gamma-treated cells. By flow cytometric analyses, no decrease in expression of surface Fas antigen or p55 TNFR was observed in differentiated U937 cells. Cell surface expression of CD11b was seen only when differentiation was induced with VD or RA but not with IFN-gamma. The growth of VD- or RA-treated cells was retarded but IFN-gamma-treated cells were prolific. These findings suggest that the differentiation state differs with the inducer and that the cellular response to apoptotic induction is closely related to the state including the cell cycle.

Effects of luteinizing hormone and dibutyryl adenosine 3',5'-monophosphate in cultured granulosa cells from polycystic ovaries.

The present study was undertaken to assess the ability of granulosa cells from subjects with normal and polycystic ovaries (PCO) to secrete progesterone throughout a 10-day culture period. LH levels in serum and follicular fluid from PCO patients were significantly (P less than 0.001) higher than those in normal subjects. In the absence of LH, progesterone secretion by granulosa cells cultured from PCO follicles did not differ significantly from that of cells from normal early and midfollicular phase follicles. Granulosa cells cultured from follicles from normal subjects in the early and midfollicular phases responded to LH (100 ng/ml) with an 8- to 20-fold increase in progesterone production. In contrast, LH increased progesterone production to a much lesser extent (up to 4-fold) in cells from the ovaries of patients with PCO. Progesterone secretion by granulosa cells from normal ovaries in response to LH diminished as intrafollicular endogenous progesterone and LH levels increased. Cells from PCO follicles cultured with (Bu)2cAMP (100 micrograms/ml) secreted progesterone in quantities comparable to those secreted by (Bu)2cAMP-stimulated normal ovaries in the early and midfollicular phases. These data demonstrate the discrepancy between the ability of granulosa cells from PCO and normal follicles to secrete progesterone in response to stimulation by LH and (Bu)2cAMP. These results suggest that in women with PCO, the persistent elevation of follicular LH may lead to impaired progesterone production in response to exogenous LH.

Levels of inhibin-F activity and steroids in human follicular fluid from normal women and women with polycystic ovarian disease.

To examine inhibin-F activity (FSH-suppressing activity) in human follicular fluid of polycystic ovary (PCO) patients, 13 follicles from 5 documented PCO patients and an additional 31 follicles from normal women in the follicular phase of the menstrual cycle were sampled, and inhibin-F activity was measured in rat anterior pituitary cell cultures. Inhibin-F activity was measured in follicular fluid after stripping steroids from the fluids using treatment with dextran and activated charcoal. Estrogen, progesterone, and delta 4-androstenedione in the follicular fluid were also determined by RIA. Estrogen and progesterone levels in follicular fluid from PCO follicles 3.9 +/- 0.34 mm in diameter were comparable with those in follicular fluid obtained from viable follicles (which had a delta 4-androstenedione to estrogen ratio of 10 or less) from normal women. delta 4-Androstenedione levels in PCO follicles were higher (P less than 0.01) than those in viable and atretic follicles of normal women. Inhibin-F levels in PCO follicles were comparable to those in viable follicles, but significantly greater (P less than 0.01) than levels in atretic follicles of normal women. If inhibin-F levels in both atretic and viable follicles of normal women were pooled, the levels were less (P less than 0.05) compared to the level in PCO follicular fluid. As an additional control, follicular fluid was collected from 90 follicles of normal women throughout the menstrual cycle, and follicular size was determined as well as inhibin-F and steroid content. Small follicles less than 8 mm; (comparable in size to the PCO follicles examined) obtained at each stage represented 79%, 24%, 0%, and 94% of the total follicles obtained in the early to midfollicular, late follicular, preovulatory, and luteal phases of the cycle, respectively. Inhibin-F activity in the fluids of these follicles was less than that in PCO follicular fluid. The average inhibin content of all of the small normal follicles was 197 +/- 34 U/10 microliter, which was significantly less (P less than 0.05) than the level in PCO follicles (332 +/- 13 U/10 microliters). These data represent the first observation of inhibin-F activity in PCO follicular fluid and suggest the possibility of involvement of inhibin-F in bringing about low or normal basal levels of FSH in the presence of elevated basal LH levels often observed in PCO patients.

A preliminary study of free amino acids in the postmortem temporal cortex from Alzheimer-type dementia patients.

Concentrations of free amino acids were measured in the temporal cortex of postmortem brains from four histologically verified cases with Alzheimer-type dementia (ATD) and eight histologically normal controls. The concentrations of taurine, glutamate, and gamma-aminobutyric acid, which are all neurotransmitter candidates, were significantly lower in the ATD brains than in the controls. These findings suggest that the involvement of amino acid neurons in ATD cannot be ruled out.

New pathologic observations in juvenile onset parkinsonism with dystonia.

A patient with hereditary juvenile onset parkinsonism with dystonia died at age 39. There were Lewy bodies and regionally selective neuronal damage in the substantia nigra pars compacta. These changes closely resemble those seen in Parkinson's disease, and emphasize the selective vulnerability of the ventral tier of the pars compacta in these degenerations.

Current comprehensive therapy of habitual abortion.

Habitual abortion is sometimes an incurable pathogenetic state. Even more serious to both the patient and gynecologist is that the incidence of repeated reproductive wastage is higher in subsequent pregnancies. We registered more than 1,000 women with repeated spontaneous abortions in this clinic for 5 years and analyzed their pathogenesis for the wastages by several different approaches. One hundred twenty-four women were diagnosed as having a congenital uterine abnormality. Metroplasty was performed in 50 patients, resulting in 82% successful subsequent pregnancies, whereas more than 98% of the pregnancies had terminated in spontaneous abortion before the operation. By measuring anticardiolipin antibody (ACA) and lupus anticoagulant (LA), the present study diagnosed several cases as antiphospholipid syndrome, and successfully pregnant patients were observed and treated with appropriate medications. Fifty-five partners of 54 couples had either a chromosomal abnormality or normal variants, showing a high incidence of spontaneous abortion in each type of abnormality. In the other 311 cases, female partners were treated by the immunotherapy of the husbands' lymphocytes in pre- and postgestational periods. Among them, 200 women became pregnant, and 147 cases successfully maintained the pregnancy. These studies indicate that early diagnosis through comprehensive examinations is essential for effective treatment of curable fetal loss in human reproduction.

Free amino acids in post-mortem cerebral cortices from patients with Alzheimer-type dementia.

Concentrations of free amino acids were measured in the cerebral cortices of post-mortem brains from 5 histologically verified cases of Alzheimer-type dementia (ATD) and 8 histologically normal controls. The concentration of glutamate in the ATD brains was significantly lower in the superior frontal, orbital, cingulate and inferior temporal cortices when compared with the control brains. The concentrations of taurine and gamma-aminobutyric acid in the ATD brains were significantly lower in the inferior temporal cortex. These findings suggest that amino acid neurons could be involved in ATD.

A behavioural pharmacological study on intracerebroventricularly administered CCK-8 related peptides in mice.

The sulfated form of cholecystokinin octapeptide (CCK-8) and ceruletide (CER), but not their non-sulfated forms of CCK-4, significantly decreased the rates of locomotor activity and rearing during the first 10 min test session 10 min after intracerebroventricular (ICV) administration at doses more than 25 and 3.125 mg, respectively. CER-S antagonized methylphenidate-induced hypermotility after ICV administration at a dose of 800 ng. Plasma levels of CER-like immunoreactivity (CER-LI) measured at 120 min after subcutaneous injection, when the locomotor suppressive activity induced by 100 and 200 micrograms was no longer observed, were similar to or much higher than that 30 min after ICV administration at a dose of 800ng, suggesting that the effects of ICV CER-S are not mediated by a peripheral redistribution. These findings indicate that (1) the structural requirement for the locomotor suppressive activity is sulfated tyrosine residue; (2) the behavioural effects of ICV-administered CCK-8-S and CER-S are due to their central actions and mediated by the/inhibition of the central dopamine (DA) function; and (3) CCK-8-S in the brain is functionally associated with the central DA system.

Determination of serotonin in microdialysis samples from rat brain by microbore column liquid chromatography with post-column derivatization and fluorescence detection.

The present paper describes a new method for on-line determination of 5-HT in brain microdialysates from awake rats by microbore column liquid chromatography with post-column derivatization and fluorescence detection. The derivatization reagent contained 1 mM benzylamine and 0.5 mM potassium hexacyanoferrate (III), both dissolved in a mixture of acetonitrile and 25 mM borate buffer (pH 11.0) (1:1, v/v). The limit of detection (S/N=3) for 5-HT was 0.5 fmol/20 microl. The samples were injected every 20 min onto a microbore column packed with C18 silica gel. The method exhibits an excellent stability over the periods of at least 12-24 h. The basal levels of 5-HT from 25 awake rats were 7.10+/-1.06 fmol/20 microl in the dorsal hippocampus and 4.64+/-0.91 fmol/20 microl (mean+/-SD) in the striatum. The 5-HT release increased to about 1500% during the perfusion with 100 mM K(+) containing Ringer solution or it was reduced to 60 or 40% during the perfusion with 1 microM tetrodotoxin or calcium free Ringer, respectively. The new method can be used to monitor extracellular 5-HT following acute systemic drug administration.

Extrahypophyseal distribution of alpha-melanocyte stimulating hormone (alpha-MSH)-like immunoreactivity in postmortem brains from normal subjects and Alzheimer-type dementia patients.

Using a sensitive double-antibody solid-phase enzyme immunoassay method alpha-melanocyte stimulating hormone-like immunoreactivity (alpha-MSH-LI) was measured in 21 regions of postmortem brains from 8 normal subjects and 5 patients with Alzheimer-type dementia (ATD). In the brains from the normal subjects, the highest concentration of alpha-MSH-LI was found in the hypothalamus. Relatively high concentration were also measured in the locus coeruleus, substantia innominata, substantia nigra, amygdala and medial nucleus of thalamus. alpha-MSH-LI in other regions was approximately 1/100 of the hypothalamic content. This data is consistent with the existence of alpha-MSH in extrahypophyseal regions and indicates its regional distribution in the human brain. In the Alzheimer brains, although the temporal cortex and hippocampus had normal concentrations of alpha-MSH-LI, the cingulate cortex, caudate and substantia nigra showed significantly lower concentrations of alpha-MSH-LI than those of the control brains. This data suggests that further studies of alpha-MSH content in a larger number of ATD brains would be useful.

Loss of dendritic synapses in the medial amygdala associated with kindling.

Kindling stimulation was given in the basolateral amygdala (BLA), the septal area or the corpus callosum in the right hemisphere of adult rats. The density of dendritic synapses was electron microscopically studied in the medial amygdaloid nucleus (MAN) ipsi- and contralateral to the stimulation side. The number of dendritic synapses was markedly decreased in both sides of the MAN of 3 groups of kindled rats. Such reduction occurred in both dendritic shaft and spine synapses. The most remarkable decrease was obtained in the BLA kindling. These results suggest that a decrease of synapses may provide a morphological basis for kindling.

Calbindin-immunoreactive cholinergic neurones in the nucleus basalis of Meynert in Alzheimer-type dementia.

An antibody to the calcium binding protein, calbindin D28K (CaBP), was used to study the number and size of CaBP-immunoreactive neurones in the nucleus basalis of Meynert (nbM) of postmortem human brains from neurologically normal controls and from patients with neuropathologically diagnosed Alzheimer-type dementia (ATD). In controls, almost all the large neurones and their processes in the nbM were CaBP immunoreactive. Compared to neurologically normal controls the number of CaBP-immunoreactive neurones in the nbM in patients dying with ATD was significantly reduced and there was a clear loss of the majority of CaBP immunoreactive neurones. The few remaining nbM CaBP immunoreactive neurones in the ATD cases were smaller than those in the neurologically normal controls. Double-staining experiments revealed that many of the nbM CaBP-immunoreactive neurones contained choline acetyltransferase immunoreactivity, so that CaBP is an alternative marker for the nbM cholinergic neurones in the human fore-brain. These findings suggest that a disturbance in calcium homeostasis may be a possible factor contributing to the loss of these cholinergic/CaBP-containing neurones.

Cell membrane changes in brains manifesting senile plaques: an immunohistochemical study of GM1 membranous ganglioside.

To investigate structural changes in cell membranes of the Alzheimer-type dementia (ATD) brain, we immunostained for GM1 ganglioside which is a major component of the cell membrane. Our results have shown that astrocytic membranes and senile plaques (SPs) have the same immunoreactivity against the monoclonal anti-ganglioside GM1 antibody. Moreover, the astrocytic processes within the SPs were altered and their abnormal membranes seemed to contribute to the formation of SPs.

Parvalbumin-immunoreactive neurons in the cortex in Pick's disease.

Parvalbumin (a calcium-binding protein)-immunoreactive (PV-Ir) neurons in the cerebral cortex were examined in 20 postmortem brains obtained from elderly controls and patients with Pick's disease (PD). The type of PV-Ir neurons and their distribution in control and PD brains were similar. The number of PV-Ir neurons in PD brains did not differ significantly from that in the control brains either. These findings suggested that PV-Ir neurons in the cortex are not affected in PD brains. A significant loss of PV-Ir neurons has already been reported in brains obtained from patients with Alzheimer-type dementia (ATD), and the present results suggest the possibility that the damage of PV-Ir neurons might be comparatively selective for ATD brains.

Clomiphene citrate and its effects upon ovulation and estrogen.

PIP: This paper reports a clinical evaluation of the mechanism of action of clomiphene citrate and describes selection of the most responsive patients. Patients were 121 women, aged 21-37 years, who desired pregnancy. Their infertility was diagnosed as being due to anovulation. Primary amenorrhea or special endocrine disorders were not present. All the women who had no vaginal bleeding for more than 2 months were diagnosed amenorrhea and treated with 65 mg of progesterone capronate intramuscularly. They were then divided into two subgroups on the basis of the presence or absence of vaginal bleeding within 2 weeks. Clinical studies included: basal body temperature charts; daily vaginal smears evaluated by the ink acidophilic stain index (ISI); cervical mucus evaluated by amount, spresence of spinnbarkeit, and ferning; 24-hour urines examined for estrogen and total gonadotropic activity; and a pregnanediol determination. Each group received daily 50 mg doses of clomiphene citrate for 5 days. Estrogen inhibiting effect of the drug was suggested by vaginal cytology and the disappearance of ferning and decrease in quantity of cervical mucus. However, the excretion of the total urinary estrogen was increased in ovulatory cases (81 of the 121 patients). In 17 patients having no bleeding within 2 weeks after progesterone injection no ovulation could be induced. In patients with withdrawal flow 54 of 70 achieved ovulation. Of 37 patients with previous anovulatory bleeding 27 achieved ovulation. There were 11 of the 121 who became pregnant. In those with early ovulation the antiestrogen effect is believed to be in the hypothalamus and pituitary bringing about the estrogen surge and stimulating LH secretion. In those with later ovulation the antiestrogenic effect increased FSH secretion followed by ovulation. The type of patient most likely to respond to clomiphene citrate is one with nearly normal pituitary-gonadal axis. Inducing withdrawal bleeding with progesterone in those amenarrheic patients who are to be treated with clomiphene citrate is believed to be a good method of anticipating the result of the treatment.^ieng

Immunoreactive luteinizing hormone-releasing hormone in the seminal plasma and human semen parameters.

A luteinizing hormone-releasing hormone (LH-RH)-like substance has been detected in human seminal plasma by a radioimmunoassay (RIA) with a highly specific anti-LH-RH antiserum. The seminal samples--not only the plasma itself but also the sample extracted by an acid/alcohol method--showed satisfactory displacement curves in our RIA system. The relationship between fertility and the LH-RH values in the seminal plasma was studied by comparing the peptide levels with sperm concentration and motility. By these two parameters, 103 samples were divided into four groups. In the low-concentration groups (oligozoospermic patients), the hormonal concentrations differed significantly (P less than 0.05) between those specimens demonstrating good and poor motility. These data suggest that this immunoreactive LH-RH may play a role in human spermatogenesis.