Evaluating ring-tailed lemurs (Lemur catta) from southwestern Madagascar for a genetic population bottleneck

In light of historical and recent anthropogenic influences on Malagasy primate populations, in this study ring-tailed lemur (Lemur catta) samples from two sites in southwestern Madagascar, Beza Mahafaly Special Reserve (BMSR) and Tsimanampetsotsa National Park (TNP), were evaluated for the genetic signature of a population bottleneck. A total of 45 individuals (20 from BMSR and 25 from TNP) were genotyped at seven microsatellite loci. Three methods were used to evaluate these populations for evidence of a historical bottleneck: M-ratio, mode-shift, and heterozygosity excess tests. Three mutation models were used for heterozygosity excess tests: the stepwise mutation model (SMM), two-phase model (TPM), and infinite allele model (IAM). M-ratio estimations indicated a potential bottleneck in both populations under some conditions. Although mode-shift tests did not strongly indicate a population bottleneck in the recent historical past when samples from all individuals were included, a female-only analysis indicated a potential bottleneck in TNP. Heterozygosity excess was indicated under two of the three mutation models (IAM and TPM), with TNP showing stronger evidence of heterozygosity excess than BMSR. Taken together, these results suggest that a bottleneck may have occurred among L. catta in southwestern Madagascar in the recent past. Given knowledge of how current major stochastic climatic events and human-induced change can negatively impact extant lemur populations, it is reasonable that comparable events in the historical past could have caused a population bottleneck. This evaluation additionally functions to highlight the continuing environmental and anthropogenic challenges faced by lemurs in southwestern Madagascar.     

Calpain from rat intestinal epithelial cells: Age-dependent dynamics during cell differentiation

Micromolar and millimolar Ca²⁺-requiring neutral protease (calpain I and calpain II) along with their endogenous inhibitor calpastatin were isolated and partially purified from the same preparation of rat intestinal epithelial cells. Calpain I and II were partially purified by 1300 and 900-fold with 57 and 53 per cent yield, respectively. The optimum assay conditions revealed pH 7.5, 20 min incubation at 25° C and 0.24% casein substrate for both calpains. The optimum calcium concentration obtained for calpain I and II were 25 M and 4 mM, respectively. Distribution of rat intestinal epithelial cells calpain I and II along with calpastatin during cell differentiation stages in weanling to senescence age were studied. Calpain I in weanling rats was in an increasing order from villus to crypt regions. Adult rats indicated well expressed consistent calpain I throughout the differentiation stages. Whereas, significant lowering towards crypt region cells were evident in old rats. Calpain II in weanling and adult rats was found to be consistent throughout the differentiation stages. Old animals revealed an increasing trend from villus to crypt region with insignificant activity present in upper villus cells. Concomitantly, different concentrations of calpastatin were observed throughout the differentiation stages in all the age groups. Moreover, the levels of calpains exceeded that of calpastatin in most of the epithelial cell populations during developmental stages. In addition to casein, intestinal epithelial cell membranes were found to be equally good substrates for calpains. Proteolytic susceptibility of weanling, adult and old rat membrane proteins varied significantly all along the ageing process in rats. Simultaneous age-dependent calpastatin response were also evident. Taken together the results obtained provided strong evidence that calpain plays significant role in rat intestinal cell differentiation and ageing process with calpastatin as its specific regulatory protein.Abbreviations DEAE-cellulose O-(Diethylaminoethyl)-cellulose - EDTA Ethylene Diamine Tetra Acetic Acid - Tris Tris (hydroxymethyl) amino methane - KH₂PO₄ potassium dihydrogen orthophosphate - Na₂HPO₄ disodium hydrogen phosphate - CaCl₂ Calcium Chloride - TCA Trichloroacetic Acid - PMSF Phenylmethylsulfonyl Fluoride     

Molecular genetic diversity and association mapping of morphine content and agronomic traits in Turkish opium poppy (<Emphasis Type="Italic">Papaver somniferum</Emphasis>) germplasm

As the sole plant source of many potent alkaloids, opium poppy (Papaver somniferum L.) is an important medicinal crop. Nevertheless, few studies have characterized opium poppy germplasm with crop-specific molecular markers. Because Turkey is a diversity center for opium poppy, Turkish germplasm is a valuable genetic resource for association mapping studies aimed at identifying QTLs controlling morphine content and agronomic traits. In this study, the morphological diversity and molecular diversity of 103 Turkish opium poppy landraces and 15 cultivars were analyzed. Potentially useful morphological variation was observed for morphine content, plant height, and capsule index. However, the landraces exhibited limited breeding potential for stigma number, and seed and straw yields. Both morphological and molecular analyses showed distinct clustering of cultivars and landraces. In addition, a total of 164 SSR and 367 AFLP polymorphic loci were applied to an opium poppy association mapping panel composed of 95 opium poppy landraces which were grown for two seasons. One SSR and three AFLP loci were found to be significantly associated with morphine content (P < 0.01 and LD value (r ²) = 0.10–0.32), and six SSR and 14 AFLP loci were significantly associated with five agronomic traits (plant height, stigma number, capsule index, and seed and straw yields) (P < 0.01 and LD value (r ²) = 0.08–0.35). This is the first report of association mapping in this crop. The identified markers provide initial information for marker-assisted selection of important traits in opium poppy breeding.     

Micro-organisms associated with African locust bean (Parkia filicoidea Welw) fermentation for 'dawadawa' production

Micro-organisms associated with the fermentation of 'dawadawa', a traditionally prepared food condiment, were isolated and identified. Three species of bacteria ( Bacillus subtilis, Leuconostoc mesenteroides and L. dextranicus ) were the predominant and most actively involved organisms. The other organisms did not appear to play any major role in the fermentation because they were present in relatively low numbers in the fermenting mash. The temperature and the pH increased sharply during the fermentation process.     

Micelle‐enhanced spectrofluorimetric method for quantification of entacapone in tablets and human plasma

In this paper, a simple and highly sensitive spectrofluorimetric method was developed and validated for the determination of entacapone (ETC). The proposed method is based on forming a highly fluorescent product through the reduction of ETC with Zn/HCl. The produced fluorophore exhibits strong fluorescence at λ_em 345 nm after excitation at λ_ex 240 nm. The use of fluorescence enhancers such as Tween‐80 and carboxy methyl cellulose (CMC) greatly enhanced the fluorescence of the produced fluorophore by 150% and 200%, respectively. Calibration curves showed good linear regression (r2 > 0.9998) within test ranges of 0.05–2.0 and 0.02–1.80 μg mL^?1 with lower detection limits of 1.27 × 10^?2 and 4.8 × 10^?3 μg mL^?1 and lower quantification limits of 4.21 × 10^?2 and 1.61 × 10^?2 μg mL^?1 upon using Tween‐80 and or CMC, respectively. The method was successfully applied to the analysis of ETC in its pharmaceutical formulations (either alone or in presence of other co‐formulated drugs). The results were in good agreement with those obtained using the official method. The methods were further extended to determine the drug in human plasma samples, and to study the pharmacokinetics of ETC. The paper is the first report on the spectrofluorimetric determination of entacapone.     

Recruitment of the nucleotide excision repair endonuclease XPG to sites of UV-induced dna damage depends on functional TFIIH

The structure-specific endonuclease XPG is an indispensable core protein of the nucleotide excision repair (NER) machinery. XPG cleaves the DNA strand at the 3' side of the DNA damage. XPG binding stabilizes the NER preincision complex and is essential for the 5' incision by the ERCC1/XPF endonuclease. We have studied the dynamic role of XPG in its different cellular functions in living cells. We have created mammalian cell lines that lack functional endogenous XPG and stably express enhanced green fluorescent protein (eGFP)-tagged XPG. Life cell imaging shows that in undamaged cells XPG-eGFP is uniformly distributed throughout the cell nucleus, diffuses freely, and is not stably associated with other nuclear proteins. XPG is recruited to UV-damaged DNA with a half-life of 200 s and is bound for 4 min in NER complexes. Recruitment requires functional TFIIH, although some TFIIH mutants allow slow XPG recruitment. Remarkably, binding of XPG to damaged DNA does not require the DDB2 protein, which is thought to enhance damage recognition by NER factor XPC. Together, our data present a comprehensive view of the in vivo behavior of a protein that is involved in a complex chromatin-associated process.     

The effect of seaweed composite flour on the textural properties of dough and bread

Seaweeds as food and seaweed-derived food flavors, colors, and nutrients are attracting considerable commercial attention. In the baking industries, hydrocolloids are of increasing importance as bread making improvers, where their use aims to improve dough handling properties, increase the quality of fresh bread, and extend the shelf life of stored bread. Seaweeds contain a significant amount of soluble polysaccharides and have the potential function as a source of dietary fiber. In this study, red seaweed (Kappaphycus alvarezii) powder was incorporated (2–8 %) with wheat flour and used to produce bread. The effect of seaweed composite flour on dough rheological properties and the quality of bread was investigated using various techniques. Farinograph tests were applied to determine the effect of seaweed powder on the rheological properties of wheat flour dough, while texture profile analysis (TPA) was used to measure the textural properties of dough as well as the final product. The results showed that the additions of seaweed powder (2–8 %) increased the water absorption of the dough. TPA results showed that the addition of seaweed powder decreased stickiness properties. Bread produced with seaweed composite flour showed higher values of firmness.     

Prevalence and intensity of Angiostrongylus cantonensis in freshwater snails in relation to some ecological and biological factors

The purpose of the study was to record different intermediate hosts of A. cantonensis and to determine the infection prevalence and intensity of this parasite in freshwater snails in relation to some ecological and biological factors. The study was conducted at Al-Salam irrigation Canal and Al-Abtal village (north Sinai) for one year, from March 2004 to February 2005. Thirteen species of freshwater snails of nine families were examined for A. cantonensis infection. Six species were found infected with A. cantonensis larvae. These species were L. carinatus, C. bulimoides, C. cyclostomoides, B. alexandrina, L. natalensis and M. tuberculta. The infection prevalence of A. cantonensis in the examined snails ranged from 0.63 to 2.24%. L. carinatus snail had the highest prevalence, mean abundance and mean intensity of A. cantonensis infection. Positive correlations were found between both prevalence and mean abundance of A. cantonensis and host size in L. carinatus and M. tuberculata. Negative correlations were detected between salinity and prevalence, mean abundance and mean intensity of larvae of A. cantonensis. The results demonstrated seasonal and spatial variation in the prevalence, mean abundance and mean intensity of infection among examined snails. In this study, A. cantonensis larvae were found in a wide range of freshwater snails and M. tuberculata snail was recorded as a new intermediate host for the first time. In conclusion, further investigations in other areas and controlled laboratory experiments of infection approaches are required to evaluate the possible threat of this parasite on humans.