1-14C-Linoleic Acid distrubution in various tissue lipids of guinea pigs following an oral dose

A recent study on the metabolism of 1-¹⁴C-α-linolenic acid in the guinea pig revealed that the fur had the highest specific activity of all tissues examined, 48 h after dosing. The present study investigated the pattern of tissue lipid labeling following an oral dose of 1-¹⁴C-linoleic acid after the animals had been dosed for the same time as above. Guinea pigs were fed one of two diets with a constant linoleic acid content (18% total fatty acids) and a different content of α-linolenic acid (0.3 or 17.3%) from weaning for 3 wk and 1-¹⁴C-linoleic acid was given orally to each animal for 48 h prior to sacrifice. The most highly labeled tissues (dpm/mg of linoleic acid) were liver, followed by brain, lung and spleen, heart, kidney and adrenal and intestines, in both diet groups. The liver had almost a three-fold higher specific activity than skin and fur which was more extensively labeled than the adipose and carcass. Approximately two-thirds of the label in skin plus fur was found in the fur which, because of a low lipid mass, would indicate that the fur was highly labeled. All tissues derived from animals on the diet with the low α-linolenic acid level were significantly more labeled than the tissues from the animals on the high α-linolenic acid diet, by a factor of 1.5 to 3. The phospholipid fraction was the most highly labeled fraction in the liver, free fatty acids were the most labeled fraction in skin & fur, while triacyglycerols were the most labeled in the carcass and adipose tissue. In these tissues, more than 90% of the radioactivity was found in fatty acids with 2-double bonds in the tissue lipids. These data indicate that the majority of label found in guinea pig tissues 48 h after dosing was still associated with a fatty acid fraction with 2-double bonds, which suggests there was little metabolism of linoleic acid to more highly unsaturated fatty acids in this time frame. In this study, the labeling of guinea pig tissues with linoleic acid, 48 h after dosing, was quite different from the labeling with α-linolenic acid reported previously. The retention of the administered radioactivity from ¹⁴C-linoleic acid in the whole body lipids was 1.6 times higher in the group fed the low α-linolenic acid diet (diet contained a total of 1.8 g PUFA/100 g diet)compared with the group fed the high α-linolenic acid diet (diet contained 3.6 g PUFA/100 g diet). The lack of retention of ¹⁴C-labeled lipids in the whole body would be consistent with an increased rate of β-oxidation of the labeled fatty acid on the diet rich in PUFA, a result supported by other studies using direct measurement of labeled carbon dioxide.

Effect of Alpha-Linolenic acid on retnal function in mammals

The retinal function of several different species is compromised by a dietary deficiency of α-linolenic acid. A review of the literature revealed that diets containing less than O. 1 gIl OOg diet as α-linolenic acid could not sustain retinal docosahexaenoic acid levels over a prolonged period and that such diets were associated with a reduced response of the retina to light. In these studies the median α-linoleinic acid intake of control animals was 1.25g/100g diet. A study on the comparative ability of dietary α-linolenic acid and dietary docosahexaenoic acid to provide for retinal docosahexaenoic acid in the guinea pig found that α-linolenic acid was only approximately 10% as effective as docosahexaenoic acid in this regard.

Studies of the human lymphocyte P2Z receptor and its activation of phospholipase D

Extracellular adenosine 5′-triphosphate (ATP) is an agonist for the P2Z receptor of human leukaemic lymphocytes and opens a Ca ²⁺-selective ion channel, which also conducts Ba²⁺, Sr²⁺ and the small fluorescent dye, ethidium⁺. A wide range of receptor agonists, many of which raise cytosolic [Ca²⁺] activate phospholipase D (PLD). In the present study, it was shown that both ATP and 3′-O-(4-benzoylbenzoyl)-ATP (BzATP) stimulated PLD activity in a concentration-dependent manner, and the inhibitory effects of suramin, oxidised ATP, extracellular Na⁺ and Mg²⁺ suggested that the effect of these agonists is mediated by P2Z receptors. The role of divalent cations in ATP-stimulated PLD activity was investigated. Several agonists (eg ATP, thapsigargin, ionomycin) stimulated a rise in cytosolic [Ca²⁺] in human lymphocytes, but only ATP and ionomycin stimulated PLD activity. When Ca²⁺ influx was prevented by EGTA, the majority of ATP-stimulated and all of ionomycin-stimulated PLD activity was inhibited. Preloading cells with the Ca²⁺ chelator, BAPTA, reduced cytosolic [Ca²⁺] and, paradoxically, ATP-stimulated PLD activity was potentiated. ATP-stimulated PLD activity was supported by both Ba²⁺ and Sr²⁺ when they were substituted for extracellular Ca²⁺. Furthermore, both ATP-stimulated PLD activity and ATP-stimulated ¹³³Ba²⁺ influx showed a linear dependence on extracellular [Ba²⁺]. Thus it was concluded that ATP stimulated PLD activity in direct proportion to the influx of divalent cations through the P2Z ion channel and this PLD activity was insensitive to changes in bulk cytosolic [Ca²⁺]. The calmodulin (Ca²⁺/CaM) inhibitor, trifluoperazine (TFP) inhibited ionomycin- and ATP-stimulated PLD activity and ATP-stimulated apoptosis, but had no effect on PLD activity already activated by ATP. However, TFP inhibited ATP-stimulated Ca²⁺, Ba²⁺ and ethidium⁺ fluxes, at concentrations below those which inhibit Ca²⁺/CaM, suggesting that TFP inhibits the P2Z receptor. Similarly, the isoquinolinesulphonamide, KN-62, a selective inhibitor of Ca²⁺/CaM-dependent protein kinase II (CaMKII), also prevented ATP-stimulated apoptosis, but had no effect on pre-activated PLD. In addition, KN-62, and an analogue, KN-04, which has no effect on CaMKII, potently inhibited ATP-stimulated Ba²⁺ influx (IC₅₀ 12.7 ± 1.5 and 17.3 ± 2.7 nM, respectively), ATP-stimulated ethidium⁺ uptake (IC₅₀ 13.1 ± 2.6 and 37.2 ± 8.9 nM, respectively), ATP-stimulated phospholipase D activity (50% inhibition 5.9 ± 1.2 and 9.7 ± 2.8 nM, respectively) and ATP-induced shedding of the surface adhesion molecule, L-selectin (IC₅₀ 31.5 ± 4.5 and 78.7 ± 10.8 nM, respectively). They did not inhibit phorbol ester- or ionomycin-stimulated PLD activity or phorbol ester-induced L-selectin shedding. Neither KN-62 nor KN-04 (both 500 nM) have any effect on UTP-stimulated Ca²⁺ transients in fura-2-loaded human neutrophils, a response which is mediated by the P2Y₂ receptor, neither did they inhibit ATP-stimulated contractile responses mediated by the P2X₁ receptor of guinea pig urinary bladder. Thus, KN-62 and KN-04 are almost equipotent as P2Z inhibitors with IC₅₀s in the nanomolar, indicating that their actions cannot be due to CaMKII inhibition, but rather that they are potent and direct inhibitors of the P2Z receptor. Extracellular ATP-induced shedding of L-selectin from lymphocytes into the medium is a Ca²⁺-independent response. L-selectin is either cleaved by a metalloproteinase or a PLD with specificity for glycosylphosphatidylinositol (GPI). The novel hydroxamic acid-based zinc chelator, Ro-31-9790 blocks ATP-induced L-selectin shedding, but was without effect on ATP-induced Ba²⁺ influx or ATP-stimulated PLD activity. Furthermore, another zinc chelator, 1,10-phenanthroline, an inhibitor of a GPI-PLD, potentiated rather than inhibited ATP-stimulated PLD activity, suggesting that ATP-induced L-selectin shedding and ATP-stimulated PLD activity are independent of each other. Although extracellular ATP is the natural ligand for the lymphocyte P2Z receptor, it is less potent than BzATP in stimulating Ba²⁺ influx. Concentration-response curves for BzATP- and ATP-stimulated ethidium⁺ influx gave EC₅₀s 15.4 ± 1.4 µM and 85.6 ± 8.8 µM, respectively. The maximal response to ATP was only 69.8 ± 1.9% of that for BzATP. Hill coefficients were 3.17 ± 0.24 and 2.09 ± 0.45 for BzATP and ATP respectively, suggesting greater positive cooperativity for BzATP than for ATP in opening the P2Z-operated ion channel. A rank order of agonist potency of BzATP > ATP = 2MeSATP > ATPγS was observed for agonist-stimulated ethidium⁺ influx, while maximal influxes followed a rank order of BzATP > ATP > 2MeSATP > ATPγS. When ATP (300 -1000 µM) was added simultaneously with 30 µM BzATP (EC₉₀), it reduced both ethidium⁺ and Ba²⁺ fluxes by 30 - 40% relative to values observed with BzATP alone. KN-62, previously shown to be a specific inhibitor of the lymphocyte P2Z receptor, was a less potent antagonist of BzATP-induced fluxes than ATP, when maximal concentrations of both agonists (50 and 500 µM respectively) were used. However, when BzATP (18 µM) was used at a concentration equiactive with a maximally effective ATP concentration, KN-62 showed the same inhibitory potency for both agonists. The ecto-ATPase antagonist, ARL-67156, inhibited both ATP- and BzATP-stimulated Ba²⁺ influx, suggesting that the lower efficacy of ATP compared with BzATP was not due to preferential hydrolysis of ATP. Thus, the natural ligand, ATP, is a partial agonist for the P2Z receptor while BzATP is a full agonist. Moreover the competitive studies show that only a single class of P2-receptor (P2Z class) is expressed on human leukaemic lymphocytes. Both ATP- and BzATP-stimulated PLD activity were significantly inhibited (P < 0.05) when cells were suspended in iso-osmotic choline Cl medium. Choline⁺ was found to be a permeant for the P2Z ion channel, since ATP induced a large uptake of [¹⁴C]choline⁺ (60 to 150 µmol/ml intracellular water) during a 5 min incubation, which remained in the cells for several hours, and ATP was used to load cells with these levels of choline⁺. Intracellular choline⁺ inhibited ATP-, BzATP-, PMA- and ionomycin-stimulated PLD activity. Brief exposure of lymphocytes to ATP increased the subsequent basal rate of ethidium⁺ uptake, and this was prevented by intracellular choline⁺. It is proposed that P2Z-mediated Ca²⁺ influx in lymphocytes activates PLD leading to significantly changes of the phospholipid composition of the plasma membrane, which subsequently produces a permeability lesion, which in turn contributes to cell death.

Progesterone and testosterone in combination act in the hypothalamus of castrated rams to regulate the secretion of LH

We tested the hypotheses that progesterone enhances the negative feedback actions of testosterone in rams and that this occurs through actions at the hypothalamus. In the first part of this study, blood samples were collected every 10 min for 12 h before and after 7 days of treatment (i.m.) of castrated Romney Marsh rams (n=5 per group) with vehicle, progesterone (4 mg/12 h), testosterone (4 mg/12 h) or a combination of progesterone (4 mg/12 h) and testosterone (4 mg/12 h). In the second part of this study the brains of four gonad-intact Romney Marsh rams were collected, the hypothalamus was sectioned and in situ hybridisation of mRNA for progesterone receptors conducted. After 7 days of treatment with vehicle or progesterone or testosterone alone, there were no changes in the secretion of LH. In contrast, treatment with a combination of progesterone and testosterone resulted in a significant (P<0.01, repeated measures ANOVA) decrease in mean plasma concentrations of LH, the number of LH pulses per hour and the pre-LH pulse nadir and a significant (P<0.01) increase in the inter-LH pulse interval. We found cells containing mRNA for progesterone receptors throughout the hypothalamus, including the preoptic area (where most GnRH neurons are located in sheep), the periventricular, ventromedial and arcuate nuclei and the bed nucleus of the stria terminalis. This study shows that progesterone is capable of acting centrally with testosterone to suppress the secretion of LH in castrated rams and that cells containing mRNA for progesterone receptors are located in the hypothalamus of rams in the vicinity of GnRH neurons.

Observations on ponticocythereis tricristata (Brady, 1880) from the Admiralty Islands, Papua New Guinea and comments on quaternary ostracod evolution within the sw Pacific Ocean

The ostracod species originally described as Cythere tricristata Brady, 1880 from the Admiralty Islands, Papua New Guinea, appears to belong to the SW Pacific and Australasian genus Ponticocythereis McKenzie, 1967 (sensu Warne & Whatley 1996). This interpretation is based on the presence of some posterior pointing scale-like spines on the carapace surface of this species. SEM images of the type material for Ponticocythereis tricristara n. comb., which are presented here for the first time, enable the clear differentiation of this species from the very similar Ponticocythereis ichthyoderma (Brady, 1890), Ponticocythereis quadriserialis (Brady, 1890) and Ponticocythereis laingensis (Wouters, 1981). As a consequence of the subdued manifestation of scale-like or blade-like spines on adult specimens of P. tricristata, this species closely resembles juvenile rather than adult specimens of some other Ponticocythereis species. This ontogenetic/phylogenetic relationship suggests that paedomorphic processes were significant in the Quaternary evolutionary radiation of Ponticocythereis species within tropical SW Pacific and Australasian regions.

Disrupting assumptions about vernacular education in Papua New Guinea

During 1999 and 2000, the author was employed by the Papua New Guinea Department of Education. Part of her duties at the Papua New Guinea Education Institute was to deliver in-service professional development programs to teachers who were implementing the country’s new curriculum. A focus of this curriculum is the use of two languages in early education; it also responds to the notion of the “bridging years” when children in Papua New Guinea develop skills and knowledge in two cultures and two languages. In this article, the author casts a critical gaze on the implications of developing literacy technologies for oral languages in Papua New Guinea. She uses examples drawn from the in-service course she developed for practicing teachers, as well as referring to theoretical understandings of the importance of literacy practices being learned in social and cultural contexts.

Export-led growth hypothesis: evidence from Papua New Guinea and Fiji

Purpose - This paper aims to examine the export-led growth hypothesis for Fiji and Papua New Guinea (PNG).

Design/methodology/approach – The paper investigates the export-led growth hypothesis for Fiji and PNG who have been facing dismal economic growth performances over the last couple of decades.

Findings – Findings of the study suggest that for Fiji there is evidence of export-led growth in the long-run, while for PNG there is evidence of export-led growth in the short-run.

Originality/value – The findings of this paper have important messages for policy makers given that export sectors in both countries investigated are underdeveloped due mainly to a sustained period of political instability.

Effects of dietary conjugated linoleic acid on haematological and humoral responses in the grower pig

The effects of conjugated linoleic acid (CLA) on the levels of total serum leucocytes, granulocytes including neutrophils, basophils, and eosinophils, as well as on monocytes and leucocytes were measured in pigs selected from a clean (minimal disease) herd. Thirty pigs were fed different rates of dietary CLA (0, 1.25, 2.5, 5.0, 7.5, and 10.0 g CLA-55/kg diet) for 8 weeks. Blood samples were collected at the end of the study for assessment of haematological and humoral responses to CLA supplementation. No difference in total white blood cells including the neutrophil, monocyte, and lymphocyte counts was observed among different dietary groups. A dose-dependent reduction (P = 0.02) in eosinophil concentrations suggests that CLA exerts anti-inflammatory activities. A 2-fold increase in the level of basophils was recorded in pigs fed lower levels of CLA (1.25 and 2.5 g CLA/kg diet) but the levels decreased gradually (P = 0.05) and were below the detection limit at the highest rate (10 g/kg) of CLA supplementation. The level of IgG was reduced by over 50% in CLA-fed pigs (P < 0.001), although the response was quadratic in nature (P < 0.001). T-cell population analysis showed that CD4+ cells tended (P = 0.06) to be reduced linearly with increasing inclusion of CLA in the diet. Our results suggest that dietary CLA modulates haematological and humoral responses in a dose-dependent manner.

Dietary conjugated linoleic acid improves carcass leanness without altering meat quality in the growing pig

One constraint facing the pig industry is that ad libitum feeding can often result in high levels of body fat and technologies which can reduce the ratio of lean to fat deposition in the pig are continually being explored. Conjugated linoleic acids have been shown to decrease body fat content in pigs. Therefore, the aim of this study was to determine whether dietary conjugated linoleic acids supplementation has any effect on meat quality and carcass characteristics in finisher pigs. Sixty female crossbred (Large White × Landrace) pigs (average initial weight 56.6 ± 1.9 kg and average initial P2 backfat 11.4 ± 1.3 mm) were used in the present study. Pigs were individually housed and randomly allocated to 1 of 6 dietary treatments: 0, 0.125, 0.25, 0.50, 0.75 or 1.0% (w/w) of conjugated linoleic acids-55. The wheat-based diets were formulated to contain 14.3 MJ DE and 9.3 g available lysine per kg and were fed ad libitum for 8 weeks. Pigs were slaughtered and meat quality was determined on the longissimus thoracis using standard techniques. Dietary conjugated linoleic acids reduced subcutaneous back fat in a linear manner with effects being most pronounced in the middle back fat layer. There was also a linear (P<0.001) decrease in intramuscular fat with increasing dietary conjugated linoleic acids supplementation. However, there was no effect of conjugated linoleic acids on subjective measures of marbling of the loin. Also, loin muscle ultimate pH (P = 0.94), lightness values (P = 0.46) subjective colour scores (P = 0.79), cooking loss (P = 0.71), drip loss (P = 0.40), shear force (P = 0.61) and subjective measures of wetness/firmness (P = 0.19) were unaffected. Dietary conjugated linoleic acids did not alter oxidation, as measured by the level of TBARs at day 1 post-slaughter (P = 0.38) or after 9 days of simulated retail display (P = 0.35). These data confirm that dietary conjugated linoleic acids can improve carcass quality by decreasing back fat depths without having any detrimental effects on meat quality.