Characterizing sleep disorders in an autism-specific collection of electronic health records.

OBJECTIVE/BACKGROUND: Sleep problems are common in people on the autism spectrum. This study reviews one detailed approach to querying the electronic health record (EHR) in a large tertiary care center. PATIENTS/METHODS: We developed methods for identifying people on the autism spectrum and defined their sleep problems using the key words, "sleep" or "melatonin", or International Classification of Diseases (ICD) codes. We examined treatment responses of these individuals to melatonin supplementation. RESULTS: Sleep problems were documented in 86% of patients with ages ranging from 6 to 30 years old. Our specific keyword search yielded more patients with sleep diagnoses than ICD codes alone. About two-thirds of patients who received melatonin supplementation reported benefit from its use. CONCLUSIONS: Our study provides a framework for using deidentified medical records to characterize sleep, a common co-occurring condition, in people on the autism spectrum. Using specific keywords could be helpful in future work that queries the EHR.

Effects of prepartum controlled-energy wheat straw and grass hay diets supplemented with starch or sugar on periparturient dairy cow performance and lipid metabolism.

This study examined the effects of a forage source [wheat straw (WS) versus grass hay (GH)] prepartum and supplemental carbohydrate source [corn (dry feed; DF) versus molasses (liquid feed; LF)] on pre- and postpartum intake, digestibility, selective particle consumption, milk yield, and lipid metabolism. The objectives were to determine if forage or pre- and postpartum supplement alters periparturient intake, energy balance, and milk yield. Sixty (n=15) multiparous dairy cows were used in a randomized complete block design with a 2 × 2 factorial arrangement of treatments to compare WS versus GH diets supplemented with either DF or LF. Dietary treatments were (1) WS prepartum + DF pre- and postpartum (WSDF), 2) WS prepartum + LF pre- and postpartum (WSLF), (3) GH prepartum + DF pre- and postpartum (GHDF), and (4) GH prepartum + LF pre- and postpartum (GHLF). Treatments began at dry-off, × before expected calving. During the prepartum phase, cows maintained dry matter intake (DMI) at 2.0% of body weight and prepartum energy balance remained positive for all treatments until calving. Prepartum GH diets had a more positive energy balance compared with WS diets. On week -5, energy balance was more positive for GHDF than for WSDF or GHLF. Energy balance for WSLF, however, was lower on week -3 and -1 than GHDF. Liquid feed decreased dry matter digestibility and increased prepartum liver triglyceride, serum nonesterified fatty acids (NEFA), and tended to increase ß-hydroxybutyrate. After calving, LF decreased DMI and energy balance, but not yield of milk or 3.5% fat-corrected milk, resulting in greater feed efficiency compared with DF. Forage did not affect postpartum DMI, but milk yield tended to be higher for WS versus GH. The DMI expressed as percentage of body weight was not affected by supplement or prepartum forage type. Cows fed WS had lower serum NEFA, higher liver glycogen, and tended to have a lower triglyceride to glycogen ratio postpartum than GH. Serum NEFA peaked on d 14 for all treatments and then declined thereafter. In postpartum diets, more particles were retained on the top screen for LF (>19.0mm) of the Penn State Particle Separator, which also tended to have more particles in the second screen (particles 19.0-8.0mm). Supplement had minimal effect on postpartum selective particle consumption. In conclusion, feeding diets containing WS resulted in lower postpartum serum NEFA, higher liver glycogen, and a tendency for greater milk production and lower liver triglyceride to glycogen than those containing GH. Liquid feed reduced postpartum DMI but not yield of milk yield or 3.5% fat-corrected milk yield, resulting in an improvement in feed efficiency. Future research should continue to investigate the use of single dry cow diet feeding strategies as they affect pre- and postpartum animal responses.

Chronic exposure to typical or atypical antipsychotics in rodents: temporal effects on central alpha7 nicotinic acetylcholine receptors.

A decrease in alpha7 nicotinic acetylcholine receptors in the hippocampus has been hypothesized to contribute to alterations in auditory gating and other behavioral impairments in schizophrenia. However, while both typical and atypical neuroleptics are routinely used in the therapeutics of schizophrenia, little is known about their effects on auditory gating or alpha7 nicotinic acetylcholine receptor expression particularly when they are administered for extended periods of time (which is common in the clinical setting). In the present study in normal rats, the residual effects of prior chronic treatment (90 or 180 days) with representative typical and atypical neuroleptics (oral haloperidol, 2.0 mg/kg/day; chlorpromazine, 10.0 mg/kg/day, risperidone, 2.5 mg/kg/day; or olanzapine, 10.0 mg/kg/day) on prepulse inhibition of the auditory gating response were investigated. The densities of alpha7 nicotinic acetylcholine receptors were subsequently measured using [125I]-alpha-bungarotoxin autoradiography. The results indicated that none of the compounds significantly altered the startle amplitude or prepulse inhibition response either during drug treatment (day 60) or after 90 or 180 days of treatment (i.e. during a drug free washout). However, prior exposure to chlorpromazine, risperidone and olanzapine for 90 days resulted in modest but significant (P<0.01) decreases in [125I]-alpha-bungarotoxin binding sites in some brain regions (e.g. posterior cortical amygdala). After 180 days of treatment, decreases in [(125I]-alpha-bungarotoxin binding ranging from approximately 12% (lateral dentate gyrus) up to 24% (e.g. CA1 hippocampal region) were evident in the risperidone group in 13 of the 36 regions analyzed while decreases associated with the other neuroleptics agents were still present, but not statistically significant. These data indicate that the commonly used atypical neuroleptic, risperidone is associated with time dependent and persistent negative effects on an important biological substrate of memory (i.e. the alpha7 nicotinic receptor), but that the magnitude of the deficits was not sufficient to impair auditory gating.

Effect of Withania somnifera on CTL activity.

The stimulatory effect of Withania somnifera on cytotoxic T lymphocytes was determined in Balb/C mice by the Winn's Neutralization assay using CTL sensitive EL4 thymoma cells as target cells. Withania somnifera showed a significant increase in CTL production both in vivo as well as in vitro. The survival time of E14 cells alone in treated animals was only 21 days and in the Withania extract treated group was 45 days with a percentage increase in life span (% ILS) of 114.28%. The survival rate of animals administered with EL4 cells (target) which were incubated with alloimmunized spleen cells (effector) from normal Balb/c mice was 33 days (% ILS 57.14%) and when this group was treated with Withania extract (20 mg/dose/animal i.p. for 10 days) their life span was increased to 53 days (% ILS 152.38%). The survival days of animals after the administration of EL4 cells incubated with alloimmunized spleen cells (effector) from Withania treated animals was 48 days (% ILS 118.18%) and when these animals continued with the Withania treatment their life span increased to 61 days (%ILS 177.27%). There was a significant increase in life span when animals were treated with EL4 cells incubated with effector cells generated by mixed lymphocyte culture of Balb/c mice and C57BL/6 mice in the presence (172.73%) and absence of Withania extract (59.09%). These studies indicate that Withania somnifera could stimulate the generation of cytotoxic T lymphocyte and it may reduce tumour growth.

Effect of Withania somnifera on cell mediated immune responses in mice.

Effect of Withania somnifera on the cellular immune responses (CMI) was studied in normal as well as tumour bearing animals. Administration of Withania extract was found to enhance the proliferation of lymphocytes, bone marrow cells and thymocytes in responses to mitogens. Both PHA and Con A mitogens along with Withania treated splenocytes, bone marrow cells and thymocytes could stimulate proliferation twice greater than the normal. Withania treated splenocytes along with the mitogen LPS (10 microg/ml) could stimulate the lymphocyte proliferation six times more than the normal. Natural killer cell activity (NK) was found to be enhanced significantly in both the normal and the tumour bearing group and it was found to be earlier than the control (48.92% cell lysis). Antibody dependent cellular cytotoxicity (ADCC) was found to be enhanced in the Withania treated group on the 9th day (65% cell lysis). An early Antibody dependent complement mediated cytotoxicity (ACC) was observed in the Withania treated group on day 13 (47% cell lysis).

Cytochrome P450 CYP2J9, a new mouse arachidonic acid omega-1 hydroxylase predominantly expressed in brain.

A cDNA encoding a new cytochrome P450 was isolated from a mouse brain library. Sequence analysis reveals that this 1,958-base pair cDNA encodes a 57-58-kDa 502-amino acid polypeptide that is 70-91% identical to CYP2J subfamily P450s and is designated CYP2J9. Recombinant CYP2J9 was co-expressed with NADPH-cytochrome P450 oxidoreductase (CYPOR) in Sf9 cells using a baculovirus system. Microsomes of CYP2J9/CYPOR-transfected cells metabolize arachidonic acid to 19-hydroxyeicosatetraenoic acid (HETE) thus CYP2J9 is enzymologically distinct from other P450s. Northern analysis reveals that CYP2J9 transcripts are present at high levels in mouse brain. Mouse brain microsomes biosynthesize 19-HETE. RNA polymerase chain reaction analysis demonstrates that CYP2J9 mRNAs are widely distributed in brain and most abundant in the cerebellum. Immunoblotting using an antibody raised against human CYP2J2 that cross-reacts with CYP2J9 detects a 56-kDa protein band that is expressed in cerebellum and other brain segments and is regulated during postnatal development. In situ hybridization of mouse brain sections with a CYP2J9-specific riboprobe and immunohistochemical staining with the anti-human CYP2J2 IgG reveals abundant CYP2J9 mRNA and protein in cerebellar Purkinje cells. Importantly, 19-HETE inhibits the activity of recombinant P/Q-type Ca(2+) channels that are known to be expressed preferentially in cerebellar Purkinje cells and are involved in triggering neurotransmitter release. Based on these data, we conclude that CYP2J9 is a developmentally regulated P450 that is abundant in brain, localized to cerebellar Purkinje cells, and active in the biosynthesis of 19-HETE, an eicosanoid that inhibits activity of P/Q-type Ca(2+) channels. We postulate that CYP2J9 arachidonic acid products play important functional roles in the brain.

Effect of Withania somnifera on DMBA induced carcinogenesis.

Administration of an extract of Withania somnifera was found to reduce two stage skin carcinogenesis induced by DMBA (dimethyl benzanthracene) and croton oil. Withania somnifera was administered at a concentration of (20 mg/dose/animal i.p.) consecutively on 5 days prior to DMBA administration and continued twice weekly for 10 weeks. After the 180th day of carcinogen administration, all of the animals developed papilloma in the control group whereas only six out of 12 animals developed papilloma in the treated group. A total of 11 papillomas were found in the control group while only six developed them in the Withania somnifera treated group. Enzyme analysis of skin and liver showed significant enhancement in antioxidant enzymes such as GSH, GST, Glutathione peroxides and Catalases in Withania somnifera treated group when compared with the control. The elevated level of lipid peroxide in the control group was significantly inhibited by Withania somnifera administration. These studies indicate that Withania somnifera could reduce the papilloma induced alterations to the antioxidant defense systems.

Effect of Withania somnifera on 20-methylcholanthrene induced fibrosarcoma.

Administration of an extract from the root of the plant Withania somnifera (20mg/dose/animal i.p) was found to inhibit the 20-methylcholanthrene induced sarcoma development in mice and increase the life span of tumour bearing animals. Administration of Withania could inhibit the lipid peroxide formation (152 nanomoles/mg protein) (P<0.01) compared with control (198 nanomoles/mg protein). Withania could increase the GSH level (7.7 micromoles/mg protein) which was lowered in control tumour bearing animals (3.96 micromoles/mg protein). GST level was also significantly increased (451 micromoles/min/mg protein) (P<0.001) in Withania treated animals compared to control animals (205 micromoles/min/mg protein). All the animals in the control group developed sarcoma by the 80th day of carcinogen administration. Only 3 animals in the Withania treated group developed sarcoma by the 105th day. In control animals the survival rate was 40% but in the Withania treated group the survival rate was 100% after 15 weeks of carcinogen treatment. These results indicate that Withania could inhibit 20-methylcholanthrene induced sarcoma development in mice.

Immunomodulatory activity of Withania somnifera.

Administration of an extract from the powdered root of the plant Withania somnifera was found to stimulate immunological activity in Babl/c mice. Treatment with five doses of Withania root extract (20 mg/dose/animal; i.p.) was found to enhance the total WBC count (17125 cells/mm(3)) on 10th day. Bone marrow cellularity (27x10(6) cells/femur) as well as alpha-esterase positive cell number (1800/4000 cells) also increased significantly (P<0.001) after the administration of Withania extract. Treatment with Withania extract along with the antigen (SRBC) produced an enhancement in the circulating antibody titre and the number of plaque forming cells (PFC) in the spleen. Maximum number of PFC (985 PFC/10(6) spleen cells) was obtained on the fourth day. Withania extract inhibited delayed type hypersentivity reaction in mice (Mantoux test). Administration of Withania extract also showed an enhancement in phagocytic activity of peritoneal macrophages (76.5 pigmented cells/200) when compared to control (31.5/200 cells) in mice. These results confirm the immunomodulatory activity of W. somnifera extract, which is a known immunomodulator in indigenous medicine.

Evaluation of a Tier I screening battery for detecting endocrine-active compounds (EACs) using the positive controls testosterone, coumestrol, progesterone, and RU486.

After previously examining 12 compounds with known endocrine activities, we have now evaluated 4 additional compounds in a Tier I screening battery for detecting endocrine-active compounds (EACs): a weak estrogen receptor (ER) agonist (coumestrol; COUM), an androgen receptor (AR) agonist (testosterone; TEST), a progesterone receptor (PR) agonist (progesterone; PROG), and a PR antagonist (mifepristone; RU486). The Tier I battery incorporates 2 short-term in vivo tests (5-day ovariectomized female battery; 15-day intact male battery) and an in vitro yeast transactivation system (YTS). The Tier I battery is designed to identify compounds that have the potential to act as agonists or antagonists to the estrogen, androgen, progesterone, or dopamine receptors; steroid biosynthesis inhibitors (aromatase, 5alpha-reductase, and testosterone biosynthesis); or compounds that alter thyroid function. In addition to the Tier I battery, a 15-day dietary restriction experiment was performed using male rats to assess confounding due to treatment-related decreases in body weight. In the Tier I female battery, TEST administration increased uterine weight, uterine stromal cell proliferation, and altered hormonal concentrations (increased serum testosterone [T] and prolactin [PRL]; and decreased serum FSH and LH). In the male battery, TEST increased accessory sex gland weights, altered hormonal concentrations (increased serum T, dihydrotestosterone [DHT], estradiol [E2], and PRL; decreased serum FSH and LH), and produced microscopic changes of the testis (Leydig cell atrophy and spermatid retention). In the YTS, TEST activated gene transcription in the yeast containing the AR or PR. In the female battery, COUM administration increased uterine weight, uterine stromal cell proliferation, and uterine epithelial cell height, and increased serum PRL concentrations. In the male battery, COUM altered hormonal concentrations (decreased serum T, DHT, E2; increased serum PRL) and, in the YTS, COUM activated gene transcription in the yeast containing the ER. In the female battery, PROG administration increased uterine weight, uterine stromal cell proliferation, and uterine epithelial cell height and altered hormonal concentrations (increased serum progesterone and decreased serum FSH and LH). In the male battery, PROG decreased epididymis and accessory sex gland weights, altered hormonal concentrations (decreased serum T, PRL, FSH, and LH; increased serum progesterone and E2), and produced microscopic changes of the testis (Leydig cell atrophy). In the YTS, PROG activated gene transcription in the yeast containing the AR or PR. In the female battery, RU486 administration increased uterine weight and decreased uterine stromal cell proliferation. In the male battery, RU486 decreased epididymis and accessory sex gland weights and increased serum FSH and LH concentrations. In the YTS, RU486 activated gene transcription in the yeast containing the ER, AR, or PR. Dietary restriction data demonstrate that confounding due to decrements in body weight are not observed when body weight decrements are 10% or less in the Tier I male battery. In addition, minimal confounding is observed at body decrements of 15% (relative liver weight, T3, and T4). Hence, compounds can be evaluated in this Tier I at levels that produce a 10% decrease in body weight without confounding of the selected endpoints. Using the responses obtained for all the endpoints in the Tier I battery, a distinct "fingerprint" was produced for each type of endocrine activity against which compounds with unknown activity can be compared. These data demonstrate that the described Tier I battery is useful for identifying EACs and they extend the compounds evaluated to 16.

Effect of Withania somnifera on cyclophosphamide-induced urotoxicity.

Administration of an extract from the plant Withania somnifera (Family: Solanaceae) (20 mg/dose/animal; i.p.) for five days along with cyclophosphamide (CTX) (1.5 mmol/kg body wt. i.p.) reduced the CTX induced urotoxicity. Morphological analysis of the bladders of the CTX-treated group showed severe inflammation and dark coloration whereas CTX along with the Withania-treated group showed normal bladder morphology. The extract was found to reduce the protein level in the serum (7.92 g/l) after 4 h of CTX treatment, which was higher in the CTX alone-administered group (11.44 g/l). Blood urea N2 level which was drastically enhanced (136.78 mg/100 ml) 2 after the CTX treatment was significantly reduced (52.08 mg/100 ml) when the animals were treated with Withania extract. Similarly the glutathione (GSH) content in both bladder (1.55 micromol/mg protein) and liver (3.76 micromol/mg protein) was enhanced significantly (P<0.001) in the Withania-treated group compared with the CTX alone-treated animals (bladder 0.5 micromol/mg protein; liver 1.2 micromol/mg protein) Histopathological analysis of the bladder of CTX alone-treated group showed severe necrotic damage where as the Withania somnifera-treated group showed normal bladder architecture.

Effect of Withania somnifera on cytokine production in normal and cyclophosphamide treated mice.

Administration of an extract from the powdered root of the plant Withania somnifera (Family: Solanaceae) enhanced the levels of Interferon gamma (IFN-gamma) (75.87 pg/ml), Interleukin-2 (IL-2) (14.16 pg/ml) and Granulocyte macrophage colony stimulating factor (GM-CSF) (49.22 pg/ml) in normal Balb/c mice. The lowered levels of IFN gamma--(30 pg/ml), IL-2 (4.5 pg/ml) and GM-CSF (19.12 pg/ml) after treatment with cyclophosphamide (CTX) was reversed by the administration of Withania somnifera extract (IFN gamma--74 pg/ml; IL-2 7.5 pg/ml; GM-CSF 35.47 pg/ml). Withania extract lowered the levels of Tumour necrosis factor alpha (TNF alpha) production. Administration of bonemarrow cells from donor mice treated with Withania extract increased the spleen nodular colonies in irradiated mice (8.33) compared to those treated with normal bonemarrow cells (3.03). The number of nodular colonies increased significantly when these animals were continued with Withania extract treatment. These results indicate the immunopotentiating and myeloprotective effect of Withania somnifera as seen from the enhancement of cytokine production and stem cell proliferation and its differentiation.

Techniques for evaluation of right ventricular relaxation rate in horses and effects of inhalant anesthetics with and without intravenous administration of calcium gluconate.

OBJECTIVES: To determine the most repeatable method for evaluating right ventricular relaxation rate in horses and to determine and compare effects of isoflurane or halothane with and without the added influence of intravenously administered calcium gluconate on right ventricular relaxation rates in horses. ANIMALS: 6 Thoroughbred horses from 2 to 4 years old. PROCEDURE: 6 models (2 for monoexponential decay with zero asymptote, 3 for monoexponential decay with variable asymptote, and 1 for biexponential decay) for determining right ventricular relaxation rate were assessed in conscious and anesthetized horses. The 2 methods yielding the most repeatable results then were used to determine right ventricular relaxation rates in horses anesthetized with isoflurane or halothane before, during, and after i.v. administration of calcium gluconate. Right ventricular pressure was measured, using a catheter-tip high-fidelity pressure transducer, and results were digitized at 500 Hz from minimum rate of change in ventricular pressure. RESULTS: 2 models that used monoexponential decay with zero asymptote repeatedly produced an estimate for relaxation rate and were used to analyze effects of anesthesia and calcium gluconate administration on relaxation rate. Isoflurane and halothane each prolonged right ventricular relaxation rate, with greater prolongation evident in halothane-anesthetized horses. Calcium gluconate attenuated the anesthesia-induced prolongation in right ventricular relaxation rate, with greater response obtained in isoflurane-anesthetized horses. CONCLUSIONS AND CLINICAL RELEVANCE: Right ventricular relaxation rate in horses is assessed best by use of a monoexponential decay model with zero asymptote and nonlinear regression. Intravenous administration of calcium gluconate to isoflurane-anesthetized horses best preserves myocardial relaxant function.

Expression of peroxisome proliferator-activated receptor gamma (PPARgamma) in human transitional bladder cancer and its role in inducing cell death.

The present study examined the expression and role of the thiazolidinedione (TZD)-activated transcription factor, peroxisome proliferator-activated receptor gamma (PPARgamma), in human bladder cancers. In situ hybridization shows that PPARgamma mRNA is highly expressed in all human transitional epithelial cell cancers (TCCa's) studied (n=11). PPARgamma was also expressed in five TCCa cell lines as determined by RNase protection assays and immunoblot. Retinoid X receptor alpha (RXRalpha), a 9-cis-retinoic acid stimulated (9-cis-RA) heterodimeric partner of PPARgamma, was also co-expressed in all TCCa tissues and cell lines. Treatment of the T24 bladder cancer cells with the TZD PPARgamma agonist troglitazone, dramatically inhibited 3H-thymidine incorporation and induced cell death. Addition of the RXRalpha ligands, 9-cis-RA or LG100268, sensitized T24 bladder cancer cells to the lethal effect of troglitazone and two other PPAR- activators, ciglitazone and 15-deoxy-delta(12,14)-PGJ2 (15dPGJ(2)). Troglitazone treatment increased expression of two cyclin-dependent kinase inhibitors, p21(WAF1/CIP1) and p16(INK4), and reduced cyclin D1 expression, consistent with G1 arrest. Troglitazone also induced an endogenous PPARgamma target gene in T24 cells, adipocyte-type fatty acid binding protein (A-FABP), the expression of which correlates with bladder cancer differentiation. In situ hybridization shows that A-FABP expression is localized to normal uroepithelial cells as well as some TCCa's. Taken together, these results demonstrate that PPARgamma is expressed in human TCCa where it may play a role in regulating TCCa differentiation and survival, thereby providing a potential target for therapy of uroepithelial cancers.

Suppressive effect of cyclophosphamide-induced toxicity by Withania somnifera extract in mice.

Administration of Withania somnifera extract (Solanaceae) was found to significantly reduce leucopenia induced by cyclophosphamide (CTX) treatment. The total WBC count on the 12th day of the CTX-treated group was 3720 cells/mm3 and that of CTX along with Withania was 6120 cells/mm3. Treatment of Withania along with CTX was found to significantly (P < 0.001) increase the bone marrow cellularity (13.1 x 10(6) cells/femur) compared to CTX alone treated group (8 x 10(6) cells/femur). Administration of Withania extract increased the number of alpha-esterase positive cells (1130/4000 cells) in the bone marrow of CTX treated animals, compared to the CTX-alone treated group (687/4000 cells). The major activity of Withania somnifera may be the stimulation of stem cell proliferation. These studies indicate that Withania somnifera could reduce the cyclophosphamide induced toxicity and its usefulness in cancer therapy.

An ongoing validation of a Tier I screening battery for detecting endocrine-active compounds (EACs).

After previously examining an estrogen receptor agonist (17beta-estradiol), several additional compounds have been evaluated in a Tier I screening battery for detecting endocrine-active compounds (EACs): an estrogen receptor antagonist (ICI-182,780, ICI), an androgen receptor antagonist (flutamide, FLUT), a testosterone biosynthesis inhibitor (ketoconazole, KETO), a 5alpha-reductase inhibitor (finasteride, FIN), and an aromatase inhibitor (anastrozole, ANA). The Tier I battery incorporates two short-term in vivo tests (a 5-day ovariectomized female battery and a 15-day intact male battery) and an in vitro yeast transactivation system (YTS). The Tier I battery is designed to identify compounds that have the potential to act as agonists or antagonists to the estrogen, androgen, progesterone, or dopamine receptors, steroid biosynthesis inhibitors (aromatase, 5alpha-reductase, and testosterone biosynthesis), or compounds that alter thyroid function. ICI administration decreased uterine estrogen and progesterone receptor number in the female battery, increased serum follicle-stimulating hormone (FSH) levels and caused spermatid retention in the male battery, and activated gene transcription in the YTS containing the estrogen receptor. FLUT administration increased uterine stromal cell proliferation in the female battery and decreased weights for all androgen-dependent tissues, induced Leydig cell hyperplasia, and caused hormonal alterations (increased testosterone (T), estradiol (E2), dihydrotestosterone (DHT), luteinizing hormone (LH), and FSH) in the male battery, and competed for binding to the androgen receptor in the YTS competition assay. In the male battery KETO decreased weights for all androgen-dependent tissues, caused hormonal alterations (decreased T and DHT and increased LH and FSH), and induced spermatid retention. FIN decreased seminal vesicle and accessory sex gland (ASG) unit weight and caused hormonal alterations (decreased DHT and increased LH, and PRL) in the male battery. KETO was judged not to affect any of the endpoints in the female battery. ANA decreased ASG unit weight and serum E2 levels in the male battery. Using the responses obtained for all the endpoints in the Tier I battery, a distinct "fingerprint" was produced for each type of endocrine activity against which compounds with unknown activity can be compared. These data demonstrate that the described Tier I battery is useful for identifying EACs.

Effects of selenium supplementation for cancer prevention in patients with carcinoma of the skin. A randomized controlled trial. Nutritional Prevention of Cancer Study Group.

OBJECTIVE: To determine whether a nutritional supplement of selenium will decrease the incidence of cancer. DESIGN: A multicenter, double-blind, randomized, placebo-controlled cancer prevention trial. SETTING: Seven dermatology clinics in the eastern United States. PATIENTS: A total of 1312 patients (mean age, 63 years; range, 18-80 years) with a history of basal cell or squamous cell carcinomas of the skin were randomized from 1983 through 1991. Patients were treated for a mean (SD) of 4.5 (2.8) years and had a total follow-up of 6.4 (2.0) years. INTERVENTIONS: Oral administration of 200 microg of selenium per day or placebo. MAIN OUTCOME MEASURES: The primary end points for the trial were the incidences of basal and squamous cell carcinomas of the skin. The secondary end points, established in 1990, were all-cause mortality and total cancer mortality, total cancer incidence, and the incidences of lung, prostate, and colorectal cancers. RESULTS: After a total follow-up of 8271 person-years, selenium treatment did not significantly affect the incidence of basal cell or squamous cell skin cancer. There were 377 new cases of basal cell skin cancer among patients in the selenium group and 350 cases among the control group (relative risk [RR], 1.10; 95% confidence interval [CI], 0.95-1.28), and 218 new squamous cell skin cancers in the selenium group and 190 cases among the controls (RR, 1.14; 95% CI, 0.93-1.39). Analysis of secondary end points revealed that, compared with controls, patients treated with selenium had a nonsignificant reduction in all-cause mortality (108 deaths in the selenium group and 129 deaths in the control group [RR; 0.83; 95% CI, 0.63-1.08]) and significant reductions in total cancer mortality (29 deaths in the selenium treatment group and 57 deaths in controls [RR, 0.50; 95% CI, 0.31-0.80]), total cancer incidence (77 cancers in the selenium group and 119 in controls [RR, 0.63; 95% CI, 0.47-0.85]), and incidences of lung, colorectal, and prostate cancers. Primarily because of the apparent reductions in total cancer mortality and total cancer incidence in the selenium group, the blinded phase of the trial was stopped early. No cases of selenium toxicity occurred. CONCLUSIONS: Selenium treatment did not protect against development of basal or squamous cell carcinomas of the skin. However, results from secondary end-point analyses support the hypothesis that supplemental selenium may reduce the incidence of, and mortality from, carcinomas of several sites. These effects of selenium require confirmation in an independent trial of appropriate design before new public health recommendations regarding selenium supplementation can be made

Enhanced diallyl trisulfide has in vitro synergy with amphotericin B against Cryptococcus neoformans.

Although amphotericin B remains the drug of choice for systemic fungal infections, its use is limited by considerable side effects. In The Peoples' Republic of China, commercial Allium sativum derived compounds are widely used as an antifungal drug to treat systemic fungal infections. To evaluate the scientific merit of using A. sativum derived compounds as antifungal agents, we studied a Chinese commercial preparation, allitridium. This preparation contained mainly diallyl trisulfide as confirmed by high performance liquid chromatography. Allitridium, with and without amphotericin B, was tested to determine its efficacy in killing three isolates of Cryptococcus neoformans. The minimum inhibitory concentration of the commercial preparation was 50 micrograms/ml and the minimum fungicidal concentration was 100 micrograms/ml against 1 x 10(5) organisms of C. neoformans. In addition, the commercial preparation was shown to be synergistic with amphotericin B in the in vitro killing of C. neoformans. This study demonstrates that diallyl trisulfide and other polysulfides possess potent in vitro fungicidal effects and their activity is synergistic with amphotericin B. These observations lend laboratory support for the treatment of cryptococcal infections with both amphotericin B and the Chinese commercial preparation.

Growth hormone response to baclofen: a comparison of 10-mg and 20-mg doses in healthy men.

Growth hormone (GH) response and baclofen levels were measured in seven healthy adult men following a 10-mg and a 20-mg dose of oral baclofen (gamma-aminobutyric acidB agonist) to determine the preferred dose in baclofen challenge studies. Multivariate analysis of variance (ANOVA) with repeated measures revealed no differences between the doses. However, when a univariate ANOVA with repeated measures was performed for each dose, the 10-mg dose showed no significant GH response over time, whereas the 20-mg dose showed a significant GH response over time. The average delta GH (change in GH from baseline) was 7.84 ng/ml (SD = 10.17) for the 10-mg dose and 3.34 ng/ml (SD = 3.64) for the 20-mg dose. The variability in the delta GH response to the 10-mg dose was significantly greater than the response to the 20-mg dose of baclofen. This variance in GH response was not explained by the differences in serum baclofen levels. Thus, a 20-mg baclofen dose appears to be preferable to a 10 mg-dose in baclofen challenge studies.

The Chinese herbal remedy, T2, inhibits mitogen-induced cytokine gene transcription by T cells, but not initial signal transduction.

T2, an extract of Tripterygium wilfordii Hook F, has been reported to be effective in the treatment of a variety of autoimmune diseases, including rheumatoid arthritis. Previous studies have shown that T2 inhibited mitogen- or antigen-induced proliferation of human peripheral blood T cells and B cells, IL-2 production by T cells and Ig production by B cells. In contrast, T2 did not affect monocyte functions, such as IL-1 production and antigen presentation. The current studies sought to localize the immunosuppressive action of T2 more precisely. Results show that T2 prevented [3H]-uridine uptake by mitogen-stimulated T cells and arrested them in the early GI phase of the cell cycle. The inhibitory effects of T2 could be partially overcome by costimulating PHA activated T cells with PMA and completely nullified by costimulation with PMA plus a monoclonal antibody to CD28. Moreover, T2 had no effect on expression of IL-2R or the transferrin receptor (CD71), but inhibited production of a number of cytokines, including IL-2 and IFN-gamma by activated T cells. T2 suppressed IL-2 mRNA levels, but not IL-2R mRNA levels, in activated T cells. T2-mediated inhibition reflected suppression of IL-2 gene transcription as indicated by suppression of the expression of a reporter gene driven by the IL-2 promoter. T2 had little inhibitory effect on either IL-2 gene expression or cell cycle progression when added after initial mitogenic stimulation, indicating that an early step in the cascade of activation events was inhibited. However, initial activation events including protein tyrosine phosphorylation, the generation of diacylglycerol, IP3, and the translocation of protein kinase C were not inhibited by T2. Moreover, T2 did not inhibit the phosphatase activity of calcineurin. These results have localized the effect of T2 to a step in the T cell activation cascade after initial second messenger generation, tyrosine phosphorylation and protein kinase activation, but before IL-2 gene transcription.