The anomeric mixture of some O-galactolipid derivatives is more toxic against cancer cells than either anomer alone

The anomeric mixture of a series of O-galactolipid derivatives is revealed to be more toxic against several cancer cell lines than their either single component with the pure α- or β-configuration. This interesting phenomenon has been confirmed on pairs of synthesized O-galactosyl anomers bearing length-varied alkyl chains at the lipid end. Furthermore, the most potent mixture was determined inoffensive to a normal cell line tested.     

Docosahexaenoic acid and 17 beta-estradiol co-treatment is more effective than 17 beta-estradiol alone in maintaining bone post-ovariectomy

Bone-protective effects of combined treatment with long chain polyunsaturated fatty acids (LCPUFAs) and estrogenic compounds following ovariectomy have previously been reported. Recent evidence suggests the n-3 LCPUFA docosahexaenoic acid (DHA, 22:6n-3) is particularly bone-protective. The aim of this study was to determine whether combined treatment with DHA and estrogenic compounds has a beneficial effect on bone mass in ovariectomized (OVX) rats. Rats were randomized into 9 groups and either ovariectomized (8 groups) or sham-operated (1 group). Using a 2 x 4 factorial design approach, OVX animals received either no estrogenic compound, genistein (20 mg/kg body weight/day), daidzein, (20 mg/kg body weight/day) or 17 beta-estradiol (1 microg/day) with or without DHA (0.5 g/kg body weight/day) for 18 weeks. Bone mineral content (BMC), area (BA), and density (BMD), plasma osteocalcin and IL-6 concentrations, and red blood cell (RBC) fatty acid composition were measured. Femur BMC was significantly greater in animals treated with DHA or 17 beta-estradiol than in ovariectomized controls. Plasma carboxylated osteocalcin was significantly higher in DHA-treated animals and total osteocalcin significantly lower in 17 beta-estradiol-treated animals compared with ovariectomized controls. There were significant interactions between treatment with estrogenic compounds and DHA for femur BMC, plasma IL-6 concentration, and RBC fatty acid composition. Combined treatment with 17beta-estradiol+DHA was more effective than either treatment alone at preserving femur BMC and lowering circulating concentrations of pro-inflammatory IL-6. The percentage of n-3 LCPUFAs in RBCs was significantly greater in animals receiving 17 beta-estradiol+DHA compared with either treatment alone. There was no beneficial effect of combined DHA and phytoestrogen treatment on bone. Results from this study raise the possibility that co-treatment with 17 beta-estradiol and DHA may allow a lower dose of 17 beta-estradiol to be used to provide the same bone-protective effects as when 17 beta-estradiol is administered alone.     

Sleep timing is more important than sleep length or quality for medical school performance

Overwhelming evidence supports the importance of sleep for memory consolidation. Medical students are often deprived of sufficient sleep due to large amounts of clinical duties and university load, we therefore investigated how study and sleep habits influence university performance. We performed a questionnaire-based study with 31 medical students of the University of Munich (second and third clinical semesters; surgery and internal medicine). The students kept a diary (in 30-min bins) on their daily schedules (times when they studied by themselves, attended classes, slept, worked on their thesis, or worked to earn money). The project design involved three 2-wk periods (A: during the semester; B: directly before the exam period—pre-exam; C: during the subsequent semester break). Besides the diaries, students completed once questionnaires about their sleep quality (Pittsburgh Sleep Quality Index [PSQI]), their chronotype (Munich Chronotype Questionnaire [MCTQ]), and their academic history (previous grades, including the previously achieved preclinical board exam [PBE]). Analysis revealed significant correlations between the actual sleep behavior during the semester (MS_diary; mid-sleep point averaged from the sleep diaries) during the pre-exam period and the achieved grade (p = 0.002) as well as between the grades of the currently taken exam and the PBE (p = 0.002). A regression analysis with MS_diary pre-exam and PBE as predictors in a model explained 42.7% of the variance of the exam grade (effect size 0.745). Interestingly, MS_diary—especially during the pre-exam period—was the strongest predictor for the currently achieved grade, along with the preclinical board exam as a covariate, whereas the chronotype did not significantly influence the exam grade.     

Enhancing pollination is more effective than increased conventional agriculture inputs for improving watermelon yields

Agricultural practices to improve yields in small‐scale farms in Africa usually focus on improving growing conditions for the crops by applying fertilizers, irrigation, and/or pesticides. This may, however, have limited effect on yield if the availability of effective pollinators is too low. In this study, we established an experiment to test whether soil fertility, soil moisture, and/or pollination was limiting watermelon (Citrullus lanatus) yields in Northern Tanzania. We subjected the experimental field to common farming practices while we treated selected plants with extrafertilizer applications, increased irrigation and/or extra pollination in a three‐way factorial experiment. One week before harvest, we assessed yield from each plant, quantified as the number of mature fruits and their weights. We also assessed fruit shape since this may affect the market price. For the first fruit ripening on each plant, we also assessed sugar content (brix) and flesh color as measures of fruit quality for human consumption. Extra pollination significantly increased the probability of a plant producing a second fruit of a size the farmer could sell at the market, and also the fruit sugar content, whereas additional fertilizer applications or increased irrigation did not improve yields. In addition, we did not find significant effects of increased fertilizer or watering on fruit sugar, weight, or color. We concluded that, insufficient pollination is limiting watermelon yields in our experiment and we suggest that this may be a common situation in sub‐Saharan Africa. It is therefore critically important that small‐scale farmers understand the role of pollinators and understand their importance for agricultural production. Agricultural policies to improve yields in developing countries should therefore also include measures to improve pollination services by giving education and advisory services to farmers on how to develop pollinator‐friendly habitats in agricultural landscapes.     

Combination of u.v.-B. and ozone reduces pollen tube growth more than either stress alone

The rate of in vitro Nicotiana tabacum L. “Bel-W3” pollen tube growth was reduced 62 and 44%, respectively, when pollen tubes were exposed to 120 ppb ozone (O₃) for 3 hr or 300 μW/cm² ultraviolet-B (u.v.-B) radiation for 30 min. Petunia hybrida Vilm. “White Cascade” pollen tube growth was reduced 34 and 59%, respectively, upon exposure to O₃ or u.v.-B at the above doses. The combination of u.v.-B at 300 μW/cm² for 30 min, followed by O₃ at 120 ppb for 3 hr, reduced pollen tube growth by 79% for “Bel-W3” and 75% for “White Cascade”. The effect appeared to be additive, implying that different target areas may be affected by the two stressors. In the Northeast, plants are exposed to both u.v.-B and O₃ during the normal growing season. This may result in an unexpectedly higher stress on the reproductive system than had been previously suspected based on these two stressors acting individually.     

Hepatocyte-derived ApoE is more effective than non-hepatocyte-derived ApoE in remnant lipoprotein clearance

The importance of hepatocyte-derived apolipoprotein (apo) E in the clearance of remnant lipoproteins in the liver is controversial. To address this controversy, we compared remnant clearance in two mouse models in which apoE is primarily derived either from hepatocytes or from an extrahepatic source. Hypomorphic apoE mice universally express reduced levels of apoE in all tissues, with the liver remaining the primary source of apoE. This mouse model of hepatocyte-derived apoE was compared with Apoe(-/-) mice transplanted with mouse bone marrow as a model of primarily non-hepatocyte-derived apoE. Immunohistochemical analysis of liver sections revealed that only the hepatocyte-derived apoE model had detectable levels of apoE on hepatic sinusoidal surfaces. The non-hepatocyte-derived apoE model with plasma apoE levels similar to those in the hepatocyte-derived model had 2-fold more total plasma cholesterol, 4-fold more total plasma triglycerides, and 8-fold higher levels of apoB48, similar to Apoe(-/-) mice. Both the hepatocyte-derived and the non-hepatocyte-derived apoE models had delayed clearance of an infused bolus of (125)I-labeled remnants compared with wild-type mice. However, after 3 h, plasma remnants reached wild-type levels only in the hepatocyte-derived apoE model, which had accumulated 70 +/- 5% of wild-type levels of remnants in the liver while the non-hepatocyte-derived apoE model had accumulated only 38 +/- 4%. These results demonstrate the existence of a role for both hepatically derived and localized apoE in remnant clearance. This role likely represents the enrichment of remnants sequestered on hepatocyte, with hepatocyte-derived apoE, facilitating their receptor-mediated internalization.     

T4 DNA ligase is more than an effective trap of cyclized dsDNA

T4 DNA ligase is used in standard cyclization assays to trap double-stranded DNA (dsDNA) in low-probability, cyclic or highly bent conformations. The cyclization probability, deduced from the relative yield of cyclized product, can be used in conjunction with statistical mechanical models to extract the bending stiffness of dsDNA. By inserting the base analog 2-aminopurine (2-AP) at designated positions in 89bp and 94bp dsDNA fragments, we find that T4 DNA ligase can have a previously unknown effect. Specifically, we observe that addition of T4 ligase to dsDNA in proportions comparable to what is used in the cyclization assay leads to a significant increase in fluorescence from 2-AP. This effect is believed to originate from stabilization of local base-pair opening by formation of transient DNA-ligase complexes. Non-specific binding of T4 ligase to dsDNA is also confirmed using fluorescence correlation spectroscopy (FCS) experiments, which reveal a systematic reduction of dsDNA diffusivity in the presence of ligase. ATP competes with regular DNA for non-covalent binding to the T4 ligase and is found to significantly reduce DNA-ligase complexation. For short dsDNA fragments, however, the population of DNA-ligase complexes at typical ATP concentrations used in DNA cyclization studies is determined to be large enough to dominate the cyclization reaction.     

Delayed mating in tortricid leafroller species: simultaneously aging both sexes prior to mating is more detrimental to female reproductive potential than aging either sex alone

The effect of delayed mating on reproductive potential, longevity and oviposition period of female redbanded leafroller, Argyrotaenia velutinana (Walker) and Pandemis leafroller, Pandemis pyrusana Kearfott, was investigated in the laboratory. Virgin female or male moths of each species were held for 1, 2, 4, 6 or 10 days prior to pairing with one-day-old virgin conspecifics of the opposite sex. In addition, reproductive potential was assessed when both sexes of each species were aged for those periods prior to pairing. The expected reproduction of female A. velutinana was reduced by 34, 53, 71 and 81% for 2, 4, 6 and 10-day delays in female mating, respectively. For P. pyrusana, expected reproduction was reduced by 47, 74, 85 and 93% for 2, 4, 6 and 10-day delays in female mating, respectively. Increasing male age at mating in both species had a lesser effect on female reproductive output compared with increasing female age at mating. As male A. velutinana age at mating increased, the expected reproduction of female A. velutinana was reduced by 15, 45, 54 and 70% for 2, 4, 6 and 10-day delays, respectively. Comparing male P. pyrusana of various ages at mating, expected reproduction was reduced by 14, 42, 64 and 79% for 2, 4, 6 and 10-day delays in mating, respectively. The decrease in female reproduction when both sexes were aged prior to mating was higher than when either sex alone was aged prior to pairing with a one-day-old virgin of the opposite sex. The expected reproduction of female A. velutinana was reduced by 60, 83, 96 and 98% for 2, 4, 6 and 10-day delays in mating of both sexes, respectively. Only 7.5% of female eggs hatched when both sexes of A. velutinana were aged ten days prior to mating. When simultaneously aging both sexes of P. pyrusana prior to mating, expected reproduction was reduced by 71, 93, 96 and 99% for 2, 4, 6 and 10-day delays in mating, respectively. No P. pyrusana eggs hatched after a ten-day delay of mating for both sexes. For both species, female longevity increased and duration of oviposition period decreased with increasing female age at mating. Our results demonstrate that delayed mating in both females and males negatively affects female reproductive output in both species and that simultaneous aging of both sexes prior to mating has a greater effect than aging either sex alone. Our results suggest that laboratory studies that have paired aged females or aged males with conspecifics of optimal reproductive maturity have likely underestimated the effects of delayed mating on reproductive output.     

Power training is more effective than strength training for maintaining bone mineral density in postmenopausal women.

Physical exercise has a favorable impact on bones, but optimum training strategies are still under discussion. In this study, we compared the effect of slow and fast resistance exercises on various osteodensitometric parameters. Fifty-three postmenopausal women were randomly assigned to a strength training (ST) or a power training group (PT). Both groups carried out a progressive resistance training, a gymnastics session, and a home training over a period of 12 mo. During the resistance training, the ST group used slow and the PT group fast movements; otherwise there were no training differences. All subjects were supplemented with Ca and vitamin D. At baseline and after 12 mo, bone mineral density (BMD) was measured at the lumbar spine, proximal femur, and distal forearm by dual-energy X-ray absorptiometry. We also measured anthropometric data and maximum static strength. Frequency and grade of pain were assessed by questionnaire. After 12 mo, significant between-group differences were observed for BMD at the lumbar spine (P < 0.05) and the total hip (P < 0.05). Whereas the PT group maintained BMD at the spine (+0.7 +/- 2.1%, not significant) and the total hip (0.0 +/- 1.7%, not significant), the ST group lost significantly at both sites (spine: -0.9 +/- 1.9%; P < 0.05; total hip: -1.2 +/- 1.5%; P < 0.01). No significant between-group differences were observed for anthropometric data, maximum strength, BMD of the forearm, or frequency and grade of pain. These findings suggest that power training is more effective than strength training in reducing bone loss in postmenopausal women.     

Combination treatment with MEK and AKT inhibitors is more effective than each drug alone in human non-small cell lung cancer in vitro and in vivo

AZD6244 and MK2206 are targeted small-molecule drugs that inhibit MEK and AKT respectively. The efficacy of this combination in lung cancer is unknown. Our previous work showed the importance of activated AKT in mediating resistance of non-small cell lung cancer (NSCLC) to AZD6244. Thus we hypothesized that dual inhibition of both downstream MEK and AKT pathways would induce synergistic antitumor activity. In this study, we evaluated the efficacy of AZD6244 and MK2206 individually on a large panel of lung cancer cell lines. Then, we treated 28 human lung cancer cell lines with a combination of AZD6244 and MK2206 at clinically applicable drug molar ratios. The AZD6244-MK2206 combination therapy resulted in a synergistic effect on inhibition of lung cancer cell growth compared to the results of single drug treatment alone. MK2206 enhanced AZD6244-induced Bim overexpression and apoptosis in A549 and H157 cells. When we tested the combination of AZD6244 and MK2206 at ratios of 8∶1, 4∶1, 2∶1, and 1∶8, we found that the synergistic effect of the combination therapy was ratio-dependent. At ratios of 8∶1, 4∶1, and 2∶1, the drug combination consistently demonstrated synergy, whereas decreasing the ratio to 1∶8 resulted in a loss of synergy and produced an additive or antagonistic effect in most cell lines. Furthermore, the AZD6244-MK2206 combination therapy showed synergy in the suppression of A549 and H157 xenograft tumor growth and increased mean animal survival time. The AZD6244-MK2206 combination therapy resulted in effective inhibition of both p-ERK and p-AKT expression in tumor tissue. In addition, a significant increase of apoptosis was detected in tumor tissue from mice treated with AZD6244-MK2206 compared with that from the single agent treated mice. Our study suggests that the combination of AZD6244 and MK2206 has a significant synergistic effect on tumor growth in vitro and in vivo and leads to increased survival rates in mice bearing highly aggressive human lung tumors.     

WT1 peptide vaccination combined with BCG-CWS is more efficient for tumor eradication than WT1 peptide vaccination alone

A Wilms tumor gene WT1 is expressed at high levels not only in most types of leukemia but also in various types of solid tumors, including lung and breast cancer. WT1 protein has been reported to serve as a target antigen for tumor-specific immunotherapy both in vitro in human systems and in vivo in murine models. We have shown that mice immunized with WT1 peptide or WT1 cDNA could reject a challenge from WT1-expressing tumor cells (a prophylactic model). However, it was not examined whether WT1 peptide vaccination had the potency to reject tumor cells in a therapeutic setting. In the present study, we demonstrated for the first time that WT1 peptide vaccination combined with Mycobacterium bovis bacillus Calmette-Guérin cell wall skeleton (BCG-CWS) was more effective for eradication of WT1-expressing tumor cells that had been implanted into mice before vaccination (a therapeutic model) compared with WT1 peptide vaccination alone. An intradermal injection of BCG-CWS into mice, followed by that of WT1 peptide at the same site on the next day, generated WT1-specific cytotoxic T lymphocytes (CTLs) and led to rejection of WT1-expressing leukemia or lung cancer cells. These results showed that BCG-CWS, which was well known to enhance innate immunity, could enhance WT1-specific immune responses (acquired immunity) in combination with WT1 peptide vaccination. Therefore, WT1 peptide vaccination combined with BCG-CWS may be applied to cancer immunotherapy in clinical settings.H. Nakajima and K. Kawasaki contributed equally to this study.     

The sleep disorder canine narcolepsy is caused by a mutation in the hypocretin (orexin) receptor 2 gene.

Narcolepsy is a disabling sleep disorder affecting humans and animals. It is characterized by daytime sleepiness, cataplexy, and striking transitions from wakefulness into rapid eye movement (REM) sleep. In this study, we used positional cloning to identify an autosomal recessive mutation responsible for this sleep disorder in a well-established canine model. We have determined that canine narcolepsy is caused by disruption of the hypocretin (orexin) receptor 2 gene (Hcrtr2). This result identifies hypocretins as major sleep-modulating neurotransmitters and opens novel potential therapeutic approaches for narcoleptic patients.     

Amylin is more potent and more effective than glucagon in raising plasma glucose concentration in fasted, anesthetized rats.

Amylin is a 37 amino-acid peptide secreted from the pancreatic beta-cells. It has actions on carbohydrate metabolism in vivo, including elevation of blood glucose. In this study, the hyperglycemic effect of intravenous bolus injections of amylin was compared with similar injections of glucagon in 20-hour fasted rats lightly anesthetized with halothane. Administered doses ranged from 0.01 micrograms to 1000 micrograms (about 7 pmol/kg--750 nmol/kg for amylin and 8 pmol/kg--800 pmol/kg for glucagon). Control animals received an equal volume of saline. A single intravenous injection of amylin or glucagon led to an increase of plasma glucose levels, which peaked approximately at 1 hour after treatment. The calculated ED50 for amylin was 1.48 nmol whereas that for glucagon was 7.46 nmol; the maximum glucose increment was 4.3 mM for amylin, and 2.9 mM for glucagon. These results show that amylin is a more potent and more effective hyperglycemic agent than glucagon under these experimental conditions.     

Pyridoxal-aminoguanidine adduct is more effective than aminoguanidine in preventing neuropathy and cataract in diabetic rats.

We examined the ability of a pyridoxal-aminoguanidine adduct with both antiglycation and antioxidant activities in vitro to protect against neuropathy and cataract in streptozotocin-diabetic rats and compared the result with that of aminoguanidine. In vivo antiglycation and antioxidant activities were also compared between the adduct and aminoguanidine. Diabetic rats were given either of the compounds in their drinking water (9 mM) for 7 weeks. Neither compound affected body weight, blood glucose level or urine volume. The adduct, but not aminoguanidine, significantly improved motor nerve conduction velocity. The time to develop cataract was longer in adduct-treated rats than in untreated and aminoguanidine-treated rats. The increase in opacification of lenses in culture medium containing high glucose levels (55.5 mM) was more efficiently attenuated by the adduct than by aminoguanidine. Adduct and aminoguanidine similarly lowered glycated hemoglobin levels. The level of urinary 8-hydroxy-2'-deoxyguanosine, a marker of oxidative DNA damage, and the level of liver malondialdehyde plus 4-hydroxy-2-alkenals, a marker of tissue lipid peroxidation, both of which were elevated by diabetes, were significantly reduced by the adduct but not by aminoguanidine. These findings indicate that the pyridoxal-aminoguanidine adduct is superior to aminoguanidine in preventing diabetic neuropathy and cataracts, and we suggest that this may be at least partly due to the higher antioxidant activity of the former.     

Inferior retinal light exposure is more effective than superior retinal exposure in suppressing melatonin in humans

Illumination of different areas of the human retina elicits differences in acute light-induced suppression of melatonin. The aim of this study was to compare changes in plasma melatonin levels when light exposures of equal illuminance and equal photon dose were administered to superior, inferior, and full retinal fields. Nine healthy subjects participated in the study. Plexiglass eye shields were modified to permit selective exposure of the superior and inferior halves of the retinas of each subject. The Humphrey Visual Field Analyzer was used both to confirm intact full visual fields and to quantify exposure of upper and lower visual fields. On study nights, eyes were dilated, and subjects were exposed to patternless white light for 90 min between 0200 and 0330 under five conditions: (1) full retinal exposure at 200 lux, (2) full retinal exposure at 100 lux, (3) inferior retinal exposure at 200 lux, (4) superior retinal exposure at 200 lux, and (5) a dark-exposed control. Plasma melatonin levels were determined by radioimmunoassay. ANOVA demonstrated a significant effect of exposure condition (F = 5.91, p < 0.005). Post hoc Fisher PLSD tests showed significant (p < 0.05) melatonin suppression of both full retinal exposures as well as the inferior retinal exposure; however, superior retinal exposure was significantly less effective in suppressing melatonin. Furthermore, suppression with superior retinal exposure was not significantly different from that of the dark control condition. The results indicate that the inferior retina contributes more to the light-induced suppression of melatonin than the superior retina at the photon dosages tested in this study. Findings suggest a greater sensitivity or denser distribution of photoreceptors in the inferior retina are involved in light detection for the retinohypothalamic tract of humans.