Efficient inhibition of the Alzheimer's disease beta-secretase by membrane targeting

beta-Secretase plays a critical role in beta-amyloid formation and thus provides a therapeutic target for Alzheimer's disease. Inhibitor design has usually focused on active-site binding, neglecting the subcellular localization of active enzyme. We have addressed this issue by synthesizing a membrane-anchored version of a beta-secretase transition-state inhibitor by linking it to a sterol moiety. Thus, we targeted the inhibitor to active beta-secretase found in endosomes and also reduced the dimensionality of the inhibitor, increasing its local membrane concentration. This inhibitor reduced enzyme activity much more efficiently than did the free inhibitor in cultured cells and in vivo. In addition to effectively targeting beta-secretase, this strategy could also be used in designing potent drugs against other membrane protein targets.     

Gametogenesis and breeding of Ostrea edulis on the west coast of Ireland

Gametogenesis, spawning and larval production of the European flat oyster Ostrea edulis L. were studied in sublittoral beds in Ballinakill Harbour and Kilkieran Bay, Co. Galway, from April 1977 to November 1978 and from November 1978 to May 1980 respectively. Histol ogical preparations of gonads and percentages of oysters brooding larvae were used to monitor breeding.Maturation began in March/April with ripe gametes appearing in some oysters in mid-May. Incubating oysters were observed from mid-June to August/September. Spent oysters were numerous in September. Gametogenesis was at a minimum from October to February depending on the year of study. Ripeness as measured by four different parameters was observed to peak around 29 June 1977 and 4 July 1978 in Ballinakill Harbour, and on 10 July 1979 in Kilkieran Bay. The onset of maximum ripeness in both populations could be predicted using the formula:

$\text{D=}\text{∑}\text{i=1}\text{N}\text{(t}{}_{\mathrm{i}}\text{? t}{}_0$

where D is the thermal constant in degree-days, n = the number of days required to reach ripeness, t_i = temperature to which the oysters were exposed daily (°C) and t₀ is the developmental zero for gametogenesis. The mean thermal constant as calculated from the gametogenesis study was 554.52 ± 44.52 degree-days. Redevelopment of gametes began during late winter at mean seawater temperatures above approximately 7.0°C.     

Germination requirements of <Emphasis Type="Italic">Allanblackia parviflora</Emphasis> seeds and early growth of seedlings

Allanblackia parviflora A. Chev, is an indigenous fruit tree species that could be used in agroforestry systems with both environmental and economic benefits. The seed oil is of prime importance as a foreign exchange earner and is being developed as a rural based enterprise in many African countries notably Ghana, Nigeria, Cameroon and Tanzania. Methods for propagation and conservation of the species are therefore of prime importance. The seeds of the species are however difficult to germinate hindering its domestication process. Allanblackia parviflora seeds were collected from seven different populations in Ghana and were subjected to four different seed germination trials at the nursery of CSIR-Forestry Research Institute of Ghana at Fumesua, Kumasi. This was followed by studying the effect of stage of seed germination on establishment after potting. Intact seeds began germination at 7 months after sowing and at 24 months, significant (P < 0.001) variations were observed in the seed germination ability of 74 accessions. Mean seed germination ranged from 0 to 35%. Significant differences in seed germination percentages among populations (P < 0.001) and provenances (P < 0.05) were also observed. Removal of seed coat significantly (P < 0.001) enhanced seed germination. Germination percentages of seeds with seed coat removed ranged from 6 to 43% while seed germination percentages for intact seeds were less or equal to 3%. Incubation of seeds with seed coat removed in polythene bags gave an added advantage. Seeds with seed coat removed and kept in polythene bags started germinating from 2 weeks and within 10 months, 75 and 68% germination were obtained for plane and black polythene bags, respectively. Removal of germinants and potting when shoot length was at least 1 cm gave significantly higher growth (P < 0.001 for shoot height, P < 0.05 for stem diameter, P < 0.001 for number of leaves produced) compared to when only radicle had emerged. It became evident in this study that seeds should be classified as fully germinated only when shoots begin to grow.     

Planning tree species diversification in Kenya based on differences in tree species composition between farms. I. Analysis of tree uses

Concerns exist about the limited diversity of tree species in agricultural landscapes. Complete tree inventories were carried out on 201 farms from four villages in western Kenya to establish whether significant differences in tree species composition existed between farms, and if so their magnitude and implications for new introductions and plantings. Tree species composition was interpreted to encompass elements of both tree identity and abundance. Tree identity was viewed from both taxonomic and function (e.g. fruit, timber, medicine) perspectives. Novel types of ordination using the Hellinger ecological distance and polynomial Redundancy Analysis indicated wide heterogeneity between farms with respect to tree species composition. For the 12 most prevalent functions of trees, the analyses showed significant differences (p<0.05). Partitioning of variance identified that village location explained much of the differences between farms suggesting that farmers share tree species within villages more than between villages. Differences between farms were assessed on two-dimensional ordination graphs. For five important tree functions, including beverage, charcoal, construction, fodder and medicine, two species dominated the compositional differences. For these functions, diversification can be achieved by village-to-village sharing even in the absence of any new species introductions. A general process to determine the degree of tree diversity at farm and landscape levels and steps to increase it are discussed.     

Comparison of two insulin protocols for diabetic dogs undergoing cataract surgery

Objective To evaluate the effectiveness of two insulin doses to maintain an acceptable range of blood glucose concentrations (70–200 mg dL^?1) in the peri‐operative period in diabetic dogs. Animals Twenty‐four diabetic dogs with a median weight of 20.6 kg and a median age of 8 years old. Methods The dogs were randomly assigned to receive either 25 or 100% of their normal insulin dose subcutaneously on the morning of surgery. The anesthetic and feeding protocols were standardized. On the day before surgery, venous blood was collected for measurement of β‐hydroxybutyrate, cholesterol, glucose, glycosylated hemoglobin, hematocrit, total plasma protein and urea nitrogen. On the day of surgery, blood glucose concentrations were measured prior to anesthesia, prior to the start of surgery, 1 and 2 hours after beginning of surgery, 1 hour after extubation, at 16 : 00 hours and at 20 : 00 hours. β‐hydroxybutyrate concentrations were measured at 20 : 00 hours that day. At 08 : 00 hours the following day, β‐hydroxybutyrate and glucose concentrations were measured. The significance of differences between groups was tested with Wilcoxon's two‐tailed rank‐sum test, Chi‐square test and Fisher's exact test. Results There were no differences in insulin treatments, clinical signs, concurrent diseases and most clinicopathological parameters between the two groups of dogs at entry to the study. The 25% dose group had blood glucose values of 296 (102–601) mg dL^?1 at 16 : 00 hours and 429 (97–595) mg dL^?1 at 20 : 00 hours on the day of surgery. The 100% insulin dose group had lower corresponding values of 130 (55–375) mg dL^?1 (p = 0.04) and 185 (51–440) mg dL^?1 (p = 0.004). No other differences (p < 0.05) were detected between the two groups. Conclusions The administration of a full dose of insulin is only marginally advantageous for reducing glucose to normal (70–120 mg dL^?1) after anesthesia but neither dose consistently induced glycemic values in an acceptable range (70–200 mg dL^?1) or normoketonemia. Clinical relevance Blood glucose should be measured immediately before anesthesia and periodically throughout the peri‐operative period in all diabetic dogs because presurgical subcutaneous administration of 25 or 100% of the normal insulin dose resulted in unpredictable blood glucose concentrations.