Deposition of iron and beta-amyloid plaques is associated with cortical cellular damage in rabbits fed with long-term cholesterol-enriched diets

Hypercholesterolemia is a potential trigger of Alzheimer's disease, and is thought to increase brain levels of beta-amyloid (Abeta) and iron. However, animal models to address the mechanisms by which Abeta and iron accumulation may cause neuronal damage are poorly defined. To address this question, we fed adult rabbits a 1% cholesterol-enriched diet for 7 months. This diet was associated with increased regional deposition of both iron and Abeta peptide in the brain. Iron preferentially accumulated around Abeta plaques in the adjacent cortex, but was not found in the hippocampus. Co-localization of iron and Abeta was accompanied by apoptosis, DNA damage, blood-brain barrier (BBB) disruption, as well as dysregulation in the level of the iron-regulatory proteins, ferritin and heme-oxygenase-1. We further demonstrate that the cholesterol diet-induced apoptosis is mediated by the activation of the endoplasmic reticulum stress pathway, involving the down-regulation of the endoplasmic reticulum chaperones, calreticulin, grp78 and grp94, and the activation of the growth and arrest DNA damage protein, gadd153. Our results suggest that BBB damage and disturbances in iron metabolism may render the cortex more vulnerable than the hippocampus to the cholesterol-induced cellular stress.     

Impact of Trace Element Changes on Dehydroepiandrosterone Sulfate in Healthy and Diabetic States among Middle-Age and Elderly Egyptians

The aim of this study was to confirm if there is a link between the alteration in blood levels of trace elements (chromium, copper, lead, cadmium, and zinc) and dehydroepiandrosterone sulfate (DHEAS) in healthy and diabetic states. This study is the first study to test these parameters in Egyptians. The study included 150 subjects divided into the following four groups: healthy middle-aged, healthy elderly, middle-aged diabetics, and elderly diabetics. Our results revealed a statistically significant decrease in the level of DHEAS in the elderly compared to middle-aged healthy and diabetic groups (p < 0.05). There was a significant difference between the middle-aged groups with respect to zinc, copper, chromium, and cadmium levels. Zinc and copper were lower in the diabetic subjects while chromium and cadmium were higher in the same group in comparison to healthy subjects. In the elderly groups, there were significant increases in chromium and cadmium levels in diabetic subjects rather than healthy ones. There was a significant increase in the thiobarbituric acid reactive substance level in the elderly healthy and diabetic groups and a significant decrease in the glutathione level in the elderly groups. There was no correlation between the levels of trace elements and DHEAS or between the levels of DHEAS, oxidants, and antioxidants in all of the tested groups. In conclusion, only the DHEAS level was correlated with age. There was no difference between the diabetic and healthy groups with respect to the levels of trace elements, with the exception of chromium and cadmium, which suggests the effect of pollution on the pathogenesis of diabetes in Egyptians. No correlation existed between the levels of DHEAS and trace elements, oxidants, and antioxidants. Finally, we believe that there is a large regional variation in the levels of trace elements due to different environmental exposure and nutritional factors which are responsible for contradictory results regarding the pathogenesis of diseases related to alterations in the levels of trace elements.     

Physiological changes in major soldiers of Macrotermes gilvus (Isoptera: Termitidae) induced by the endoparasitoid Misotermes mindeni (Diptera: Phoridae)

The majority of true parasitoids manipulate their host’s physiology for their own benefit. In this study, we documented the physiological changes that occurred in major soldiers of the subterranean termite Macrotermes gilvus (Hagen) (Isoptera: Termitidae) parasitized by the koinobiont larval endoparasitoid Misotermes mindeni Disney and Neoh (Diptera: Phoridae). We compared the metabolic rate, body water content, body water loss rate, cuticular permeability, and desiccation tolerance between parasitized and unparasitized major soldiers. The metabolic rate of parasitized hosts was significantly higher than that of unparasitized termites. Mean total body water content of parasitized major soldiers (64.73 ± 3.26%) was significantly lower than that of unparasitized termites (71.99 ± 2.23%). Parasitized hosts also had significantly lower total body water loss rates (5.72 ± 0.06%/h) and higher cuticular permeability (49.37 ± 11.26 μg/cm/h/mmHg) than unparasitized major soldiers (6.75 ± 0.16%/h and 60.76 ± 24.98 μg/cm/h/mmHg, respectively). Parasitized major soldiers survived almost twice as long as unparasitized termites (LT₅₀ = 6.66 h and LT₅₀ = 3.40 h, respectively) and they had significantly higher tolerance to water loss compared to unparasitized termites (45.28 ± 6.79% and 32.84 ± 7.69%, respectively). Body lipid content in parasitized hosts (19.84 ± 6.27%) was significantly higher than that of unparasitized termites (6.17 ± 7.87%). Finally, parasitized hosts had a significantly lower percentage of cuticular water content than unparasitized major soldiers (10.97 ± 1.84% and 13.17 ± 2.21%, respectively). Based on these data, we conclude that the parasitism-induced physiological changes in the host are beneficial to the parasitoids as the alterations can clearly increase the parasite’s chances of survival when exposed to extreme environmental conditions and ensure that the parasitoids are able to complete their larval development successfully before the host dies.     

Botulinum neurotoxin and dendrotoxin as probes for studies on transmitter release

Acceptors for BoNT have been detected autoradiographically on the terminal membrane of motor nerves at a density of approximately 150/micron2 and shown to mediate toxin internalization, a process deemed essential for its inhibition of transmitter release. DTX, a protein with pronounced central neurotoxicity, was shown to induce convulsive states in hippocampal slices from guinea-pig. Synaptic transmission was facilitated and spontaneous epileptiform activity produced in intact cell populations. Voltage clamp analysis of hippocampal neurones revealed that DTX specifically attenuated a transient voltage-dependent K+ conductance (A-current) and this could account for the excitatory effects observed. Proteinaceous acceptors with high affinity for DTX were identified on brain synaptosomal membranes and found to contain a 65 000 Mr polypeptide. Their location in rat brain regions was established and contrasted with that of the binding sites for beta-bungarotoxin. These findings indicate the usefulness of DTX as a probe for a protein associated with one variety of K+ channel while the larger subunit of BoNT was found to interact with a membraneous component that resides at cholinergic nerve terminals and, hence, is likely to have a unique role.     

Regulation of Gβγi-Dependent PLC-β3 Activity in Smooth Muscle: Inhibitory Phosphorylation of PLC-β3 by PKA and PKG and Stimulatory Phosphorylation of Gαi-GTPase-Activating Protein RGS2 by PKG

In gastrointestinal smooth muscle, agonists that bind to G_i-coupled receptors activate preferentially PLC-β3 via Gβγ to stimulate phosphoinositide (PI) hydrolysis and generate inositol 1,4,5-trisphosphate (IP₃) leading to IP₃-dependent Ca²⁺ release and muscle contraction. In the present study, we identified the mechanism of inhibition of PLC-β3-dependent PI hydrolysis by cAMP-dependent protein kinase (PKA) and cGMP-dependent protein kinase (PKG). Cyclopentyl adenosine (CPA), an adenosine A₁ receptor agonist, caused an increase in PI hydrolysis in a concentration-dependent fashion; stimulation was blocked by expression of the carboxyl-terminal sequence of GRK2(495–689), a Gβγ-scavenging peptide, or Gα_i minigene but not Gα_q minigene. Isoproterenol and S-nitrosoglutathione (GSNO) induced phosphorylation of PLC-β3 and inhibited CPA-induced PI hydrolysis, Ca²⁺ release, and muscle contraction. The effect of isoproterenol on all three responses was inhibited by PKA inhibitor, myristoylated PKI, or AKAP inhibitor, Ht-31, whereas the effect of GSNO was selectively inhibited by PKG inhibitor, Rp-cGMPS. GSNO, but not isoproterenol, also phosphorylated Gα_i-GTPase-activating protein, RGS2, and enhanced association of Gα_i3-GTP and RGS2. The effect of GSNO on PI hydrolysis was partly reversed in cells (i) expressing constitutively active GTPase-resistant Gα_i mutant (Q204L), (ii) phosphorylation-site-deficient RGS2 mutant (S46A/S64A), or (iii) siRNA for RGS2. We conclude that PKA and PKG inhibit Gβγ_i-dependent PLC-β3 activity by direct phosphorylation of PLC-β3. PKG, but not PKA, also inhibits PI hydrolysis indirectly by a mechanism involving phosphorylation of RGS2 and its association with Gα_i-GTP. This allows RGS2 to accelerate Gα_i-GTPase activity, enhance Gαβγ_i trimer formation, and inhibit Gβγ_i-dependent PLC-β3 activity.     

Nutritional and ecological evaluation of dairy farming systems based on concentrate feeding regimes in semi-arid environments of Jordan

The objective of this study was to evaluate the nutritional and ecological aspects of feeding systems practiced under semi-arid environments in Jordan. Nine dairy farms representing the different dairy farming systems were selected for this study. Feed samples (n = 58), fecal samples (n = 108), and milk samples (n = 78) were collected from the farms and analysed for chemical composition. Feed samples were also analysed for metabolisable energy (ME) contents and in vitro organic matter digestibility according to Hohenheim-Feed-Test. Furthermore, fecal nitrogen concentration was determined to estimate in vivo organic matter digestibility. ME and nutrient intakes were calculated based on the farmer’s estimate of dry matter intake and the analysed composition of the feed ingredients. ME and nutrient intakes were compared to recommended standard values for adequate supply of ME, utilizable crude protein, rumen undegradable crude protein (RUCP), phosphorus (P), and calcium (Ca). Technology Impact Policy Impact Calculation model complemented with a partial life cycle assessment model was used to estimate greenhouse gas emissions of milk production at farm gate. The model predicts CH₄, N₂O and CO₂ gases emitted either directly or indirectly. Average daily energy corrected milk yield (ECM) was 19 kg and ranged between 11 and 27 kg. The mean of ME intake of all farms was 184 MJ/d with a range between 115 and 225 MJ/d. Intake of RUCP was lower than the standard requirements in six farms ranging between 19 and 137 g/d, was higher (32 and 93 g/d) in two farms, and matched the requirements in one farm. P intake was higher than the requirements in all farms (mean oversupply = 19 g/d) and ranged between 3 and 30 g/d. Ca intake was significantly below the requirements in small scale farms. Milk nitrogen efficiency N-_eff (milk N/intake N) varied between 19% and 28% and was mainly driven by the level of milk yield. Total CO₂ equivalent (CO₂ equ) emission ranged between 0.90 and 1.88 kg CO₂/kg ECM milk, where the enteric and manure CH₄ contributed to 52% of the total CO₂ equ emissions, followed by the indirect emissions of N₂O and the direct emissions of CO₂ gases which comprises 17% and 15%, respectively, from total CO₂ equ emissions. Emissions per kg of milk were significantly driven by the level of milk production (r² = 0.93) and of eDMI (r² = 0.88), while the total emissions were not influenced by diet composition. A difference of 16 kg ECM/d in milk yield, 9% in N-_eff and of 0.9 kg CO₂ equ/kg in ECM milk observed between low and high yielding animals. To improve the nutritional status of the animals, protein requirements have to be met. Furthermore, low price by-products with a low carbon credit should be included in the diets to replace the high proportion of imported concentrate feeds and consequently improve the economic situation of dairy farms and mitigate CO₂ equ emissions.     

Apolipoprotein M modulates erythrocyte efflux and tubular reabsorption of sphingosine-1-phosphate

Sphingosine-1-phosphate (S1P) mediates several cytoprotective functions of HDL. apoM acts as a S1P binding protein in HDL. Erythrocytes are the major source of S1P in plasma. After glomerular filtration, apoM is endocytosed in the proximal renal tubules. Human or murine HDL elicited time- and dose-dependent S1P efflux from erythrocytes. Compared with HDL of wild-type (wt) mice, S1P efflux was enhanced in the presence of HDL from apoM transgenic mice, but not diminished in the presence of HDL from apoM knockout (Apom^−/−) mice. Artificially reconstituted and apoM-free HDL also effectively induced S1P efflux from erythrocytes. S1P and apoM were not measurable in the urine of wt mice. Apom^−/− mice excreted significant amounts of S1P. apoM was detected in the urine of mice with defective tubular endocytosis because of knockout of the LDL receptor-related protein, chloride-proton exchanger ClC-5 (Clcn5^−/−), or the cysteine transporter cystinosin. Urinary levels of S1P were significantly elevated in Clcn5^−/− mice. In contrast to Apom^−/− mice, these mice showed normal plasma concentrations for apoM and S1P. In conclusion, HDL facilitates S1P efflux from erythrocytes by both apoM-dependent and apoM-independent mechanisms. Moreover, apoM facilitates tubular reabsorption of S1P from the urine, however, with no impact on S1P plasma concentrations.     

Effect of papaya (Carica papaya) leaf extract as dietary growth promoter supplement in red hybrid tilapia (Oreochromis mossambicus × Oreochromis niloticus) diet

The purpose of this experiment was to examine the potential use of Carica papaya leaf extract as a supplement to promote growth and improve feed utilization in red hybrid tilapia. Five diets were formulated containing isolipidic (80 g/kg) and isonitrogenic (350 g/kg) levels. All feeds contained similar types and amounts of raw materials but differed in the inclusion of papaya leaf extract (0, 5, 10, 20 and 40 g/kg feed). The initial size of fish used was 2.3 ± 0.01 g. Each diet was performed in triplicate tanks, and the feeding period was 12 weeks. Fish fed diet containing 2% papaya leaf extract (PLE) had the highest final weight, 31.14 ± 1.47 g, followed by 1% PLE (27.27 ± 1.75 g). These two diets (1% and 2%) were also showed significant improvements of weight gain, SGR, and feed efficiency of the red hybrid tilapia (p < 0.05). However, papaya leaf extract did not affect the HSI, VSI, PER, digestive enzymes activity, blood composition, and survival rate. Supplementing the diets with papaya leaf extract lowered serum urea. Findings of this research suggest that adding papaya leaf extract to the diet of red hybrid tilapia improves growth and feed efficiency without adversely affecting blood parameters. Therefore, an inclusion level between 1% and 2% of the papaya leaf extract is recommended as a feed additive to promote red hybrid tilapia fry growth.