The effect of phosphatidylserine administration on memory and symptoms of attention-deficit hyperactivity disorder: a randomised, double-blind, placebo-controlled clinical trial.

BACKGROUND: Attention-deficit hyperactivity disorder (ADHD) is the most commonly diagnosed behavioural disorder of childhood, affecting 3-5% of school-age children. The present study investigated whether the supplementation of soy-derived phosphatidylserine (PS), a naturally occurring phospholipid, improves ADHD symptoms in children. METHODS: Thirty six children, aged 4-14 years, who had not previously received any drug treatment related to ADHD, received placebo (n = 17) or 200 mg day(-1) PS (n = 19) for 2 months in a randomised, double-blind manner. Main outcome measures included: (i) ADHD symptoms based on DSM-IV-TR; (ii) short-term auditory memory and working memory using the Digit Span Test of the Wechsler Intelligence Scale for Children; and (iii) mental performance to visual stimuli (GO/NO GO task). RESULTS: PS supplementation resulted in significant improvements in: (i) ADHD (P < 0.01), AD (P < 0.01) and HD (P < 0.01); (ii) short-term auditory memory (P < 0.05); and (iii) inattention (differentiation and reverse differentiation, P < 0.05) and inattention and impulsivity (P < 0.05). No significant differences were observed in other measurements and in the placebo group. PS was well-tolerated and showed no adverse effects. CONCLUSIONS: PS significantly improved ADHD symptoms and short-term auditory memory in children. PS supplementation might be a safe and natural nutritional strategy for improving mental performance in young children suffering from ADHD.

Inhibitory effect of cinnamaldehyde, derived from Cinnamomi cortex, on the growth of influenza A/PR/8 virus in vitro and in vivo.

We have investigated the inhibitory effect of trans-cinnamaldehyde (CA), one of the principal constituents of essential oil derived from Cinnamomi cortex, on the growth of influenza A/PR/8 virus in vitro and in vivo. When 1-h drug treatment was initiated at various times post-infection (p.i.) in Madin-Darby canine kidney cells using a fixed dose of CA (40 microM), the maximum inhibitory effect (29.7% virus yield of control) was obtained when drug treatment was started at 3h p.i. Under the same treatment schedule, CA inhibited the virus growth in a dose-dependent manner (20-200 microM), and, at 200 microM, the virus yield was reduced to an undetectable level. RT-PCR and SDS-PAGE analyses showed that CA inhibited viral protein synthesis at the post-transcriptional level. In mice infected with the lung-adapted PR-8 virus, inhalation (50mg/cage/day) and nasal inoculation (250 microg/mouse/day) of CA significantly increased survival rates on the 8 days to 100% and 70%, respectively, in contrast to a survival rate of 20% in the untreated control group. Importantly, inhalation of CA caused virus yield reduction by 1 log in bronchoalveolar lavage fluid on day 6 after infection, compared with that of the untreated control group. These findings might provide further support to the empirical indication of Cinnamomi cortex-containing Kampo medicines for acute respiratory infectious diseases.

Expression of Tn and sialyl-Tn antigens in the neoplastic transformation of uterine cervical epithelial cells.

The expression of simple mucin-type carbohydrate antigens, Tn and sialyl-Tn antigens, was evaluated by immunohistochemical staining with monoclonal antibodies in normal squamous epithelium, dysplasia, carcinoma in situ, and invasive squamous cell carcinoma of the uterine cervix. The expression of the Tn antigen detected by HB-Tn1 and B1.1 was found in 17 (20%) and 19 (23%) of the 83 invasive carcinomas, respectively, but was not found in the 36 normal squamous epithelia, 22 severe dysplasias, or 24 carcinomas in situ. The sialyl-Tn antigen was detected by HB-STn1 and TKH-2 in 14 (64%) and 11 (50%) of the 22 severe dysplasias, 13 (54%) and 10 (42%) of the 24 carcinomas in situ and 48 (58%) and 42 (51%) of the 83 invasive carcinomas, respectively, but was completely absent in 36 normal squamous epithelia. Coexpression of the sialyl-Tn antigen was observed in 89% of the cases expressing the Tn antigen. No significant difference was observed between the immunoreactivities of the antigens in the metastatic lymph nodes and primary tumors. No correlation was found between the expression of each antigen and clinical state, histologic type, depth of invasion, parametrial spread, lymphatic and vessel permeation, lymph node metastasis, or 5-year survival rate. The expression of Tn and sialyl-Tn demonstrates a specific change in the neoplastic progression from carcinoma in situ to invasive carcinoma and from normal to dysplasia, respectively, in squamous cell neoplastic lesions of the cervix. Tn and sialyl-Tn antigens may be useful markers for biologic investigation of neoplastic transformation in cervical squamous cell carcinoma.

Childhood acid maltase deficiency. A clinical, biochemical, and morphologic study of three patients.

Three children, including two siblings and a patient with sporadic glycogenosis type II (childhood form of acid maltase deficiency [AMD] ), were studied clinically, biochemically, and morphologically. In addition to a delay in developmental milestones and mild generalized muscle weakness, nasal vocalization and an electromyographic finding of abnormal insertion voltage, followed by pseudomyotonic discharge, were assumed to be characteristic diagnostic findings for the childhood form of AMD. Since the neutral maltase activity was highest in the muscle biopsy specimen from the patient with the least severe weakness, the enzyme may play a role in reducing muscle involvement. Selective type 2A fiber atrophy and type 2B fiber deficiency in the affected muscles were the common histochemical findings in this particular form of AMD.

An interleukin-1 binding region oligopeptide from the human type I IL-1 receptor reduces acute inflammation, delayed hypersensitivity reaction, and lethal endotoxemia in animals.

The effect of an interleukin-1 binding region oligopeptide from the interleukin-1 receptor on various inflammatory responses was investigated in animal models. A synthetic peptide (KICIRIQIS) corresponding to 86-93 of the extracellular domain of the human type I interleukin-1 receptor was used. Carrageenan-induced rat paw edema, a model of acute inflammation, was dose dependently suppressed by intraperitoneal administration of the peptide. The delayed hypersensitivity reaction to sheep red cells was diminished by pretreatment of mice with the peptide at a relatively high dose. In a murine lethal endotoxemia model, animals treated with the interleukin-1 receptor peptide (10 mg/kg x 4) showed significantly better survival than vehicle-treated animals when the peptide was administered from 20 minutes after lipopolysaccharide injection. Improved survival was accompanied by suppression of lipopolysaccharide-induced production of colony-stimulating factor, although the peptide did not improve hypoglycemia. These findings suggest that the interleukin-1 receptor peptide may be a potential treatment for various inflammatory processes.

Efficacy of Choto-san on vascular dementia and the protective effect of the hooks and stems of Uncaria sinensis on glutamate-induced neuronal death.

Two different multicenter studies on the efficacy of Choto-san on patients with vascular dementia, one a well-controlled but non-double blind (60 patients), the other a double-blind controlled study (139 patients), were performed. In the well-controlled study, Choto-san was superior in global improvement rating, utility rating and improvement of subjective symptoms, psychiatric symptoms and disturbance in daily living activities. In the double-blind study, with more objective criteria than the well-controlled study, Choto-san was also superior in global improvement rating, utility rating and improvement of subjective symptoms, psychiatric symptoms and disturbance in daily living activities. These results suggest that Choto-san is effective in the treatment of vascular dementia. Uncaria sinensis (OLIV.) HAVIL. (US) is the main medicinal plant composing Choto-san. Glutamate-induced cell death of cultured cerebellar granule cells was protected by the application of water extract of US in a dose-dependent manner, and concentrations of 10(-5) to 10(-4) g/ml had a significant effect compared to exposure to glutamate only. Further, the increase of 45Ca2+ influx into cells by glutamate was also blocked by the water extract in a dose-dependent manner. These results suggest that US has a protective effect on glutamate-induced neuronal death in cultured cerebellar granule cells through the inhibition of Ca2+ influx.

Telomerase activity in malignant ovarian tumors with deregulation of cell cycle regulatory proteins.

Using a semiquantitative telomeric repeat amplification protocol assay, telomerase-positive frequencies and enzyme levels were measured. Out of 95% of 49 human ovarian tumors, the highest level of telomerase activity was observed in malignant tumors. Furthermore, by immunohistochemical staining of cell cycle regulatory proteins (pRB, p16, cyclin D1, cyclin E and p53) at the G1 checkpoint, we evaluated the relation between each protein alterations and the levels of telomerase activity. We could not demonstrate a clear relation with each molecule except for cyclin E, but suggesting that aberrant accumulation of these proteins was considered as a reason for telomerase deregulation, which may play an essential role in the pathway of telomerase regulation.

Inhibitory effect of bafilomycin A1, a specific inhibitor of vacuolar-type proton pump, on the growth of influenza A and B viruses in MDCK cells.

We studied the effect of bafilomycin A1 (Baf-A1), a novel and highly specific inhibitor for vacuolar-type proton (V-H+) pump, on the growth of influenza A and B viruses in Madin-Darby canine kidney cells. Vital fluorescence microscopic study showed that Baf-A1 induced the complete disappearance of acidified compartments such as endosomes and lysosomes both in infected and uninfected cells by the treatment with 0.1 microM inhibitor for 1 h at 37 degrees C. In addition, virus growth was inhibited when Baf-A1 was present from 1 h before infection to the end of incubation, or added within as early as 5-10 min after infection. Conversely, the virus growth was recovered in correlation with the reappearance of acidified compartments after removal of Baf-A1. These data suggest that Baf-A1-sensitive V-H+ pumps are solely responsible for the acidification of endosomes and lysosomes, and thus Baf-A1 inhibits the growth of influenza A and B viruses by affecting the acidified compartments in which low pH is essential for the uncoating process of influenza virus growth at an early stage of infection.

Inhibitory effect of Ephedrae herba, an oriental traditional medicine, on the growth of influenza A/PR/8 virus in MDCK cells.

Using several herbal extracts, we investigated whether certain Kampo medicines exert an inhibitory effect on the acidification of intracellular compartments such as endosomes and lysosomes (referred to as ELS), and thereby inhibit the growth of influenza A virus in Madin-Darby canine kidney cells. The vital fluorescence microscopic study showed that the extract of Ephedrae herba (EHext) among five herbal extracts inhibited acidification of endosomes and lysosomes in a concentration-dependent manner (100-400 microg/ml). Moreover the growth of influenza A/PR/8/34 (H1N1) (PR8) virus was inhibited when the cells were treated with EHext for 1 h immediately after infection, or treated as early as 5-10 min after infection. Conversely, virus growth resumed concomitantly with the reappearance of acidified ELS after removal of EHext. The fact that the inhibitory effect of EHext was completely or partially reversed by FeCl3, a tannin-reactive agent, strongly suggests that tannin is one of the active components in the extract.

Affinity chromatography on immobilized anhydrotrypsin: general utility for selective isolation of C-terminal peptides from protease digests of proteins.

Recently we have succeeded in the efficient isolation of the C-terminal peptides from tryptic digests of the tail sheath protein (with C-terminal Gly) and the tube protein (with C-terminal Glu) of bacteriophage T4, by taking advantage of a unique property of immobilized anhydrotrypsin, that is, a strong specific affinity for peptides containing Arg or Lys residues at their C-termini. In this study, the utility of affinity chromatography on immobilized anhydrotrypsin was further demonstrated in the cases of Streptomyces subtilisin inhibitor (as a reduced and S-carboxymethylated form, with C-terminal Phe) and alpha 1-antitrypsin (with C-terminal Lys). By subjecting a tryptic digest of the former protein and a chymotryptic digest of the latter protein to the affinity chromatography, the C-terminal peptides were specifically recovered in the breakthrough fraction and in the adsorbed fraction, respectively. It was further shown that immobilized anhydrotrypsin can also adsorb peptides with C-terminal S-aminoethyl-Cys residues and exerts adsorptive ability even toward the peptides in solution containing urea at a high concentration if appropriate precautions are taken. These findings suggest the general utility of this simple method for C-terminal peptide isolation, which is extremely helpful for studies to confirm amino acid sequences deduced from nucleotide sequences of the cDNA (or genomic DNA) of proteins.

Immobilized anhydrochymotrypsin as a biospecific affinity adsorbent for the peptides produced by chymotryptic hydrolysis.

Anhydrochymotrypsin immobilized on Sepharose specifically adsorbed various peptides containing L-tryptophan, L-tyrosine, or L-phenylalanine residues at their carboxy-termini. These peptides correspond to the specific products of chymotryptic cleavage of polypeptides. A mixture of the chymotryptic peptides once adsorbed on the column could be effectively separated by eluting them with a pH gradient. This adsorbent, on the other hand, showed no substantial affinity toward the substrate-type peptides that contain the aromatic amino acid(s) within the peptide chain, or toward the C-terminal leucine peptides. By taking advantage of this unique property of anhydrochymotrypsin-Sepharose in combination with reversed-phase high-performance liquid chromatography, we have succeeded in separating the C-terminal peptides from chymotryptic digests of reduced and S-carboxymethylated bovine insulin and from tryptic digests from reduced and S-carboxymethylated Streptomyces subtilisin inhibitor.

Comparisons of LET distributions for protons with energies between 50 and 200 MeV determined using a spherical tissue-equivalent proportional counter (TEPC) and a position-sensitive silicon spectrometer (RRMD-III).

Experiments have been performed to measure the response of a spherical tissue-equivalent proportional counter (TEPC) and a silicon-based LET spectrometer (RRMD-III) to protons with energies ranging from 50-200 MeV. This represents a large portion of the energy distribution for trapped protons encountered by astronauts in low-Earth orbit. The beam energies were obtained using plastic polycarbonate degraders with a monoenergetic beam that was extracted from a proton synchrotron. The LET spectrometer provided excellent agreement with the expected LET distribution emerging from the energy degraders. The TEPC cannot measure the LET distribution directly. However, the frequency mean value of lineal energy, y(-)(f), provided a good approximation to LET. This is in contrast to previous results for high-energy heavy ions where y(-)(f) underestimated LET, whereas the dose-averaged lineal energy, y(-)(D), provided a good approximation to LET.

Effects of progesterone on the ultrastructure of the golden hamster parathyroid gland.

The ultrastructure of the parathyroid glands in golden hamsters after administration of progesterone was investigated. In the parathyroid glands of the female hamsters after administration of progesterone, the mean serum calcium concentration was significantly higher when compared to that of the control hamsters. In the male experimental hamsters, the percentage area occupied by Golgi complexes and lipid droplets was significantly increased when compared to that of the control hamsters, respectively. In the female hamsters after administration of progesterone, the percentage area occupied by large vacuolar bodies was significantly decreased when compared to that of the control hamsters. In the male and female experimental hamsters, the mean number of secretory granules per 100 microns2 of cytoplasm showed a significant increase compared with that of the male and female control hamsters, respectively. These findings suggest that the secretory activity of the parathyroid glands may be stimulated in response to hypocalcemia induced by progesterone.

Effects of short-term treatment with CaCl2 or EDTA on the parathyroid glands in pregnant golden hamsters, with special reference to large vacuolar bodies.

The large vacuolar bodies in the parathyroid glands of pregnant golden hamsters after administration of CaCl2 or EDTA were investigated. In the parathyroid glands of the pregnant animals 15 min after administration of CaCl2, the mean serum calcium concentration was significantly high when compared to that of the control animals. In the parathyroid glands of the pregnant animals 15, 30 and 60 min after administration of EDTA, the mean serum calcium concentration was significantly low when compared to that of the control animals. In the parathyroid glands of the pregnant animals 15 min after administration of CaCl2, the percentage area occupied by large vacuolar bodies was significantly increased when compared to that of the control animals. In the parathyroid glands of the pregnant animals 15 min after administration of EDTA, the percentage area occupied by large vacuolar bodies was significantly decreased when compared to that of the control animals.

Effects of prostaglandin E2 on the ultrastructure of the golden hamster parathyroid gland.

The effects of different ages on large vacuolar bodies in the parathyroid glands of golden hamsters after administration of prostaglandin E2 (PGE2) were investigated. In the parathyroid glands of the young and senile animals 15 min and the senile animals 60 min after administration of PGE2, the mean serum calcium concentration was significantly higher when compared to that of the young and senile control animals, respectively. In the experimental adult animals 60 min after administration of PGE2, the serum calcium concentration was seen to increase. In the parathyroid glands of the young animals 15 min and the adult and senile animals 60 min after administration of PGE2, the percent area occupied by large vacuolar bodies was significantly increased when compared to that of the young, adult and senile control animals, respectively. These findings suggest that the percent area occupied by large vacuolar bodies is increased in response to hypercalcemia induced by PGE2. It is thought that in the parathyroid glands suppressed by hypercalcemia there is a relationship between the percent area occupied by large vacuolar bodies and aging.

An in vitro study on the effects of melatonin on the ultrastructure of the hamster parathyroid gland.

Isolated parathyroid glands from adult female golden hamsters were incubated on a black Millipore filter in an incubation vessel containing Ham's F-12 medium, with or without melatonin at final concentration of 10(-5) M for 1 hour. In the parathyroid glands used for in vitro treatments with melatonin, the Golgi complexes associated with a few prosecretory granules and cisternae of the rough endoplasmic reticulum showed a significant decrease, and lipid droplets and lysosomes appeared to be increased compared with those of the control parathyroid glands. These changes are considered to be induced by suppression of the synthesis of parathyroid hormone in parathyroid glands incubated in a vessel containing medium with melatonin.

Ultrastructure of the parathyroid gland of the young golden hamster after short-term treatment with ethanol.

We studied the ultrastructure of the parathyroid gland of young golden hamsters after short-term treatment with ethanol (1.5 g/kg bw or 6.0 g/kg bw). We did not find any ultrastructural changes of the parathyroid gland after administration of 1.5 g/kg ethanol. In the hamsters, 3 hours after administration of 6.0 g/kg ethanol, the mean serum calcium concentration was significantly low as compared to that of the control animals. In the parathyroid gland 1 hour after administration of 6.0 g/kg ethanol, the Golgi complexes associated with a few prosecretory granules and the volume density occupied by the Golgi complexes decreased compared with that of the control animals. In the parathyroid glands 3 hours after administration of 6.0 g/kg ethanol, the Golgi complexes decreased as compared with those of the control animals, while the large vacuolar bodies increased. These findings suggest that the cellular activity of the parathyroid gland is suppressed after short-term treatment with ethanol. Intracellular lumen was found in the parathyroid chief cells 3 hours after administration of 6.0 g/kg ethanol, and the significance of this structure is discussed.

Ascending projections to the lateral thalamic nuclei from the substantia grisea centralis in the rat: a retrograde WGA-HRP study.

Ascending projections to the lateral thalamic nuclear group from the substantia grisea centralis (SGC) were studied by injections of wheat germ agglutinin-conjugated with HRP (WGA-HRP) into the laterodorsal (LD) and lateroposterior (LP) thalamic nuclei. The present study demonstrated that the pars ventralis of the SGC at the levels of the intercollicular region and of the locus ceruleus sent fibers to both the LD and LP on both sides with homolateral predominance. Distribution pattern and morphological characteristics of the LD-projection neurons in the SGC were similar to those of SGC-LP projection cells. WGA-HRP injections into the LD or LP labeled also a considerable number of neurons in the dorsal raphe nucleus and the dorsal tegmental nucleus bilaterally with homolateral predominance, but the nucleus of Darkschewitsch contained labeled neurons only after the LD injection.

Descending pathways of the nucleus of the optic tract in the rat.

The descending pathways of the nucleus of the optic tract (pars lateralis) (NOTL) in the rat were examined in Nauta-Gygax impregnated sections and further confirmed by the horseradish peroxidase labeling technique. Following stereotactic lesions in the NOTL, visual cortex, and superior colliculus, projections which orginate in the NOTL were identified by elimination of the tectofugal and tectopetal fibers of the superior colliculus. The descending fibers from the NOTL appeared to be organized into three bundles. (1) The NOTL-contralateral NOTL fibers, which connect bilateral NOTLs through a part of the posterior commissure. (2) The NOTL-nucleus tegmenti pontis and inferior olivary nucleus fibers, which project to the ventromedial part of the nucleus tegmenti pontis and the dorsal cap of the inferior olivary nucleus, ipsilaterally to the NOTL. (3) The NOTL-pontine nuclei fibers, which terminate within the medial one-third of the pars lateralis of the ipsilateral pontine nuclei with topical relations.

Quantitative morphological changes in the superior colliculus and the parabigeminal nucleus in the bilaterally microphthalmic rat.

Quantitative morphological changes in the superior colliculus (SC) and the parabigeminal nucleus (PB) were studied in hereditary bilaterally microphthalmic rat, which lacks the optic nerve completely. Volumes of the retinorecipient superficial collicular layers of SC (SCS) to the central gray matter were decreased by 35% with respect to the normal. However, the cell density in SCS was increased as much as 150% of the normal. The stratum griseum superficiale was packed densely and irregularly with small-sized round nerve cells. The stratum opticum of the mutant rat appeared as a narrow band with few fiber components, but it contained some medium-sized polygonal neurons. No significant changes were found in the deeper layers of the microphthalmic SC. Bilaterally microphthalmic PB reduced both its volume and cellular density per unit area (about 30 and 75% of the normal, respectively). Furthermore, in contrast to the normal rat, the mutant PB could not be subdivided into the dorsal, middle and ventral subgroups.