Among dietary factors, learning and behavior are influenced not only by nutrients, but also by exposure to toxic food contaminants such as mercury that can disrupt metabolic processes and alter neuronal plasticity. Neurons lacking in plasticity are a factor in neurodevelopmental disorders such as autism and mental retardation. Essential nutrients help maintain normal neuronal plasticity. Nutritional deficiencies, including deficiencies in the long chain polyunsaturated fatty acids eicosapentaenoic acid and docosahexaenoic acid, the amino acid methionine, and the trace minerals zinc and selenium, have been shown to influence neuronal function and produce defects in neuronal plasticity, as well as impact behavior in children with attention deficit hyperactivity disorder. Nutritional deficiencies and mercury exposure have been shown to alter neuronal function and increase oxidative stress among children with autism. These dietary factors may be directly related to the development of behavior disorders and learning disabilities. Mercury, either individually or in concert with other factors, may be harmful if ingested in above average amounts or by sensitive individuals. High fructose corn syrup has been shown to contain trace amounts of mercury as a result of some manufacturing processes, and its consumption can also lead to zinc loss. Consumption of certain artificial food color additives has also been shown to lead to zinc deficiency. Dietary zinc is essential for maintaining the metabolic processes required for mercury elimination. Since high fructose corn syrup and artificial food color additives are common ingredients in many foodstuffs, their consumption should be considered in those individuals with nutritional deficits such as zinc deficiency or who are allergic or sensitive to the effects of mercury or unable to effectively metabolize and eliminate it from the body.

We examined the antioxidant activities of wax cappings from honey bee drone and worker brood infested with the parasitic mite Varroa destructor, as well as from worker brood from different queen lines. We used assays that measured two separate aspects of antioxidant activity in wax cappings, namely free radical scavenging capacity (DPPH assay) and ferric ion reduction capacity (FRAP assay). Mite infestation tended to have different effects on the antioxidant activities of wax cappings of drone and worker brood. Wax cappings from infested worker brood had higher antioxidant activities than cappings from uninfested worker brood. By contrast, antioxidant activities of cappings from uninfested drone brood did not differ from the activities of infested drone brood cappings. Different queen lines shared a similar overall pattern of higher antioxidant activities in cappings from infested worker brood than cappings from uninfested brood, but differed in the absolute magnitude of the antioxidant activities. In particular, Italian and "hygienic" queen lines had proportionally much higher FRAP wax capping activities in infested brood relative to uninfested brood (313% and 311%) than Russian queen lines (171%).

Instrumental insemination is an attractive alternative to natural mating because specific genetic crosses can be made, thus producing colonies with desired traits. However, there are conflicting reports on the quality and acceptance of instrumentally inseminated (II) queens compared to naturally mated (NM) queens. One factor that affects acceptance and retention of queens is the volatile compounds they produce. Our study compared volatile chemicals from virgin and mated honey bee queens that were either NM or II. The volatile compounds from virgin queens differed from those of mated egg-laying queens. Virgin queens produced greater relative amounts of the volatile compounds we detected (including 2-phenylethanol, n-octanal, and n-decanal) with the exception of E-β-ocimene, which was higher in mated laying queens. II and NM queens did not differ in type or relative amounts of volatile compounds. The similarities between II and NM queens indicate that the physiological changes that happen after mating and egg laying occur regardless of the mating method.

In the United States, high-fructose corn syrup (HFCS) has become a sucrose replacement for honey bees and has widespread use as a sweetener in many processed foods and beverages for human consumption. It is utilized by commercial beekeepers as a food for honey bees for several reasons: to promote brood production, after bees have been moved for commercial pollination, and when field-gathered nectar sources are scarce. Hydroxymethylfurfural (HMF) is a heat-formed contaminant and is the most noted toxin to honey bees. Currently, there are no rapid field tests that would alert beekeepers of dangerous levels of HMF in HFCS or honey. In this study, the initial levels and the rates of formation of HMF at four temperatures were evaluated in U.S.-available HFCS samples. Different HFCS brands were analyzed and compared for acidity and metal ions by inductively coupled plasma mass spectroscopy. Levels of HMF in eight HFCS products were evaluated over 35 days, and the data were fit to polynomial and exponential equations, with excellent correlations. The data can be used by beekeepers to predict HMF formation on storage. Caged bee studies were conducted to evaluate the HMF dose-response effect on bee mortality. Finally, commercial bases such as lime, potash, and caustic soda were added to neutralize hydronium ion in HMF samples, and the rates of HMF formation were compared at 45 degrees C.

Mercury cell chlor-alkali products are used to produce thousands of other products including food ingredients such as citric acid, sodium benzoate, and high fructose corn syrup. High fructose corn syrup is used in food products to enhance shelf life. A pilot study was conducted to determine if high fructose corn syrup contains mercury, a toxic metal historically used as an anti-microbial. High fructose corn syrup samples were collected from three different manufacturers and analyzed for total mercury. The samples were found to contain levels of mercury ranging from below a detection limit of 0.005 to 0.570 micrograms mercury per gram of high fructose corn syrup. Average daily consumption of high fructose corn syrup is about 50 grams per person in the United States. With respect to total mercury exposure, it may be necessary to account for this source of mercury in the diet of children and sensitive populations.

Bee pollen (pollen collected by honey bees) was collected in the high intensity ultraviolet (UV) Sonoran Desert and analyzed by the DPPH (radical 2,2-diphenyl-1-picryhydrazyl) assay and the FRAP (ferric reducing-antioxidant power) assay on six different pollen samples and in eight different water miscible solvents at 50mg/ml. The bee pollen taxa were characterized for each pollen type by acetylization of the pollen extracts followed by microscopy and comparison with a library of samples native to the Sonoran Desert. The standards (R-(+)-6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid), known as TROLOX, gallic acid and α-tocopherol (vitamin E) were analysed as standards to determine the potency of each pollen sample in the most efficient solvent. The Mimosa pollen sample displayed the highest antioxidant activity. Total polyphenolics, flavanols, flavones were determined, and the results are reported in milligrams of gallic acid, quercetin and naringenin per gram of pollen, respectively. There was good correlation between antioxidant activity and total phenolics. The order of effectiveness of the pollen samples in regard to antioxidant activity was determined and the most effective extraction solvents are discussed. Finally, solid phase micro-extraction, coupled with gas chromatography–mass spectroscopy was utilized to identify and quantify polyphenolic compounds known to have free radical scavenging activity in the pollen samples.

Purpose The purpose of this investigation was to synthesize a series of carbonate and carbamate derivatives of 4-demethylpenclomedine (DM-PEN), the major plasma non-toxic metabolite of penclomedine (PEN) seen in patients. DM-PEN has been observed to be an active antitumor agent in mouse human xenograft tumor models and non-neurotoxic in a rat model, however, activity in intracranially implanted human glioma xenograft models have not been reported. The major goal was to identify derivatives that are active in brain tumors. Methods Derivatives were prepared from DM-PEN and evaluated in vivo against human U251 glioblastoma, D54 glioblastoma and MX-1 breast tumor xenografts and mammary tumor 16/C that were implanted in the mammary fat pad or intracranially (IC). Results Carbonate and carbamate derivatives were found to be superior to DM-PEN against IC growing human glioblastoma xenografts. Conclusion The activity of the carbonates and carbamates against human tumor xenografts in vivo suggests consideration of these two series of derivatives of DM-PEN for clinical development.

Summary Essential oils were fed to honey bees to determine whether the components were absorbed into bee larvae. The oils were added in either sucrose syrup (origanum and 2-heptanone) or in a liquid protein diet (origanum oil, cinnamon oil, thymol, and 2-heptanone), because sugar and protein sources are differentially utilized in food fed to larvae of different ages. The volatiles emitted by isolated larvae from oil-supplemented colonies were sampled at three different ages (Day 4, Day 6 and Day 9) by Solid Phase Micro-Extraction (SPME) and analyzed by Gas Chromatography-Mass Spectrometry (GC-MS). The only oil volatiles recovered in larvae were the origanum oil components carvacrol and thymol; neither 2-heptanone nor the cinnamon oil components were detected in any larvae. For larvae from colonies fed oil- supplemented sugar syrup, carvacrol volatiles were emitted at higher amounts in younger larvae (Day 4) than in older larvae (Day 9). In contrast, carvacrol and thymol volatiles were detected only in older larvae (Day 6 and Day 9) in colonies reared on oil-supplemented liquid protein diet. Carvacrol was also detected in the cocoons of Day 9 larvae from colonies fed oil-supplemented liquid protein diet, but not oil-supplemented sugar syrup diet. We believe that the age-related differences in oil incorporation by bee larvae reflect the relative importance of supplemental sugars and protein as food sources for bee larvae of these ages. Supplementation in a liquid protein diet represents a more efficacious route for the incorporation of origanum oils in fifth instar bee larvae targeted for invasion by Varroa destructor mites.

The Varroa mite ( Varroa destructor) is becoming ubiquitous worldwide and is a serious threat to honey bees. The cultivation of certain food crops are at risk. The most noted acaricides against Varroa mites are tau-fluvaninate and coumaphos, but the mites are showing resistance. Since these insecticides are used in the proximity of honey, it is desirable to use natural alternatives. Monoterpenoids such as thymol and carvacrol, that are constituents of oil of thyme and oil of origanum, show promise as acaricides against the Varroa mite ( Varroa destructor), but the delivery of these compounds remains a challenge due to the low water solubility and uncontrolled release into the colony. Beta-cyclodextrin (beta-CD) inclusion complexes of thymol, oil of origanum, and carvacrol were prepared on a preparative scale. Competitive binding was studied by fluorescence spectroscopy by using 6- p-toluidinylnaphthalene-2-sulfonate as a fluorescent probe. The complexes were characterized, and the competitive binding described by (1)H and (13)C NMR spectroscopy chemical shifts. The toxicity of beta-CD and the prepared complexes in enriched sucrose syrup was studied by conducting caged honey bee ( Apis mellifera) feeding trials. After the first and second weeks of feeding, hemolymph and gut tissue samples were acquired from the caged bee study. The levels of thymol and carvacrol were quantified by solid-phase microextraction gas chromatography mass spectroscopy, using an optimized procedure we developed. High (mM) levels of thymol and carvacrol were detected in bee tissues without any imposed toxicity to the bees, in an effort to deter Varroa mites from feeding on honey bee hemolymph.

A280 4-DM-PEN is a non-neurotoxic metabolite of penclomedine (PEN) with anti-cancer properties. However, 4-DM-PEN does not readily cross the blood brain barrier and does not produce complete remissions in intracerebral (IC) gliomas. PEN did produce responses in clinical trials (gliomas - CR), but was neurotoxic and all trials ceased. DEKK-TEC and SRI have synthesized 20 carbonate and carbamate derivatives of 4-DM-PEN. Analogs prepared were from the series: 4-DM-PEN-4-OCO2-X & 4-DM-PEN-4-OCONH-X, where X = benzyl, methyl, ethyl, octyl, 4-Cl-, 4-F-, 4- & 2-nitrobenzyl, phenyl, 4-Cl-, 4-F-, 4-nitrophenyl, N-morpholino and cholesteryl groups. Anti-cancer activities were noted with both groups when administered IP daily x 5 days to SC growing MX-1 xenografts [response ranges - %ILS >50% and 20-40% CR]. However, only a carbonate, 4-demethyl-4-cholesteryloxy-penclomedine (DM-CHOC-PEN, X=cholesteryl) was active (produced CRs) vs. three IC implanted xenograft tumor (U251, D54 & MX-1) models with no weight loss. BCNU controls did not produce CRs in IC implanted glioma xenografts, such as D-54 in mice. The IC tumor responses for DM-CHOC-PEN are the platform for our interest in evaluating the latter drug as clinical treatment for 1o and 2o CNS malignancies in humans. The IC activity is in contrast to DM-PEN, which was active vs. MX-1 breast tumor xenografts growing SC in mice but did not produce CRs in IC implanted human U-251and D-54 glioblastoma multiforme xenografts and MX-1 xenograft models. DM-CHOC-PEN vs DM-PEN has improved activity (% ILS/CR) in IC implanted human xenograft models - U251 glioma: +29/25 vs 17/0, resp. and MX-1 breast cancer: +20/17 vs 12/0, resp. DM-CHOC-PEN’s acute toxicology in mice and dogs has been reported - AACR 48, abst. 5614, 2007. Mechanisms for CNS anticancer activity and clinical plans for Phase 1 trials will be discussed. Supported by NCI/SBIR grant - 5R44CA85021.

4,4'-dihydroxybenzophenone-2,4-dinitrophenylhydrazone (A-007) has recently completed a phase I clinical trial in advanced cancer with minimal toxicity, and impressive objective responses were noted. A-007 possesses three moieties that appear to have an influence on its anticancer activities: diphenylmethane, hydrazone, and dinitrophenyl. The goals of this study were to modify A-007's chemical moieties with the ultimate goal of maximizing its anticancer activity through increased planarity and introduction of functional groups. Thirty-five phenylhydrazone analogues of A-007 were synthesized and evaluated in vitro in a human primary cancer explant assay. Anticancer activities for selected analogues were also assayed for activity vs established human/murine cell lines. One-hundred-eighty-six fresh human solid tumors were used to screen for anticancer activity. Selected analogues were assayed for therapeutic indices (vs GM-CFC from bone marrow) in preparation for preclinical studies. Several polyaryl phenylhydrazones demonstrated improved cytotoxic activities by factors of 10(2)-10(3) when compared with A-007. However, the polyaryl quinone moieties of the latter analogues introduced potential toxic properties (cardiac, hematological) that do not exist with A-007.

The structure of the anticancer agent 4,4′-dihydroxybenzophenone-2,4-dinitrophenylhydrazone (A-007) has been modified through SAR and by incorporating barbituric acid, pyridine, quinoline, and alkylcarboxylic acids into A-007's moieties. Analogue anticancer activity and interacting with CD surface markers on a T-cell leukemia cell line were evaluated and the correlation between SAR and biological properties are discussed.

4,4′-Dihydroxybenzophenone-2,4-dinitrophenylhydrazone (A-007) formed stable double salts with phenothiazin-5-ium salts (2a–d), which have improved in vitro anticancer activities, as compared to A-007 alone. The stable salt between methylene blue (2a) and A-007 allowed the latter to diffuse into the dermis layers of skin. It is anticipated that these new salts will allow A-007 to penetrate into the deep lymphatic/vascular channels of the dermis, which contain metastatic cancer cells, and improve in vivo anticancer activities.

A very efficient method of preparation for 5-alkyl and 5-arylthiotetrazoles from the corresponding alkyl or aryl halides is described. The halides are first transformed into thiocyanates which further react with azide, yielding the corresponding tetrazoles with [2+3] polar cycloaddition. All synthetic transformations are performed under phase transfer catalytic conditions. The yields vary from good to excellent except for the preparation of 5-benzylthiotetrazole, where the reaction between benzyl thiocyanate and azide [2+3] cycloaddition is in competition with nucleophilic substitution, with benzyl azide as product.

The transition state structures for the addition of maleic acid anhydride and methyl phenylpropiolate to 1-phenyl-3,4-dimethylphosphole were generated with the AM1 semiempirical method. The competitive transition state structures for 1,5-phenyl rearrangement in 1-phenyl-3,4-dimethylphosphole was also optimized with AM1. The energies were evaluated with the SVWN and Becke3LYP DFT methods using a 6-31G(d) basis set. The reaction outcome based on the evaluated energies were discussed and compared with experimental observations.