Differentiation between Bacillus thuringiensis strains by gyrB PCR-Sau3Al fingerprinting

gyrB DNA fragments of seven Bacillus thuringiensis local collection family representatives were amplified by PCR and sequenced. Several differences in their corresponding sequences were evidenced. Both in silico and in vitro restriction maps of gyrB sequences and fragments respectively confirmed that EcoRI and Sau3AI could be used to differentiate between B. thuringiensis strains. However, the phylogeny analysis showed that only the gyrB PCR-Sau3AI allows a strains classification that correlates very well with that obtained on the basis of the sequences analysis. Thus, these finds show that gyrB PCR- Sau3AI digestion could be considered as an efficient, rapid, and easy method to make a distinction, not only between strains belonging to the Bacillus cereus group, but also between those belonging to B. thuringiensis.     

Impact of the natural resource of UVB on the content of vitamin D2 in oyster mushroom (Pleurotus ostreatus) under subtropical settings

Vitamin D deficiency is a pandemic problem. Non-animal source of vitamin D is obtained from edible mushrooms. Oyster mushroom (Pleurotus ostreatus) was sliced into the size of 1 cm³, 4 cm³ and 9 cm³, and treated with the sun as a natural resource of UVB under subtropical settings in Ethiopia. The content of vitamin D was measured by using high-performance liquid chromatography (HPLC). After sun treatment, there was a significant increment in the content of vitamin D₂ from nil to 67.4 ± 28.0 µg/g dry weight (DW). Based on the results of the overall pairwise comparisons, 1 cm³ size of slice group had the highest content of vitamin D₂. Duration of sun exposure, sizes of mushroom slices and moisture content were identified as determining factors for vitamin D₂ synthesis. Exposing slices of oyster mushroom to the sunlight for <30 min provides the amount that satisfies the current recommended dietary allowance (RDA) of vitamin D without any visible change in color and texture. Thus, sun treatment of oyster mushroom is an effective and economically cheap strategy in the fight against vitamin D deficiency.     

豹子花属及百合属13种25居群的核型研究

采用常规压片法对豹子花属6种11居群和百合属7种14居群进行了核型研究,并用不对称系数AI度量核型不对称性,以CVCL-CVCI散点图比较近缘类群之间的亲缘关系。结果表明： 1）除美丽豹子花 No-mocharis basilissa 为三倍体外,其余全为二倍体 2）核型在种间、居群间存在变异,特别是在随体染色体的数目和位置以及B染色体的有无上种间存在明显差别 3） CVCL-CVCI散点图显示豹子花属与百合属关系密切 4）染色体结构变异产生数量和类型不同的次缢痕,是豹子花属植物进化的主要方式。豹子花属在从其起源地和分化中心高黎贡山向四周扩散时,细胞核型方面伴随着出现了非整倍体、三倍体、 B染色体和次缢痕进化的类型.     

Effects of equine chorionic gonadotropin and type of ovulatory stimulus in a timed-AI protocol on reproductive responses in dairy cows

The objectives were to evaluate the effects of equine chorionic gonadotropin (eCG) supplementation (with or without eCG) and type of ovulatory stimulus (GnRH or ECP) on ovarian follicular dynamics, luteal function, and pregnancies per AI (P/AI) in Holstein cows receiving timed artificial insemination (TAI). On Day 0, 742 cows in a total of 782 breedings, received 2 mg of estradiol benzoate (EB) and one intravaginal progesterone (P4) insert (CIDR). On Day 8, the CIDR was removed, and all cows were given PGF2α and assigned to one of four treatments in a 2 × 2 factorial arrangement: (1) CG: GnRH 48 h later; (2) CE: ECP; (3) EG: eCG + GnRH 48 h later; (4) EE: eCG + ECP. There were significant interactions for eCG × ovulatory stimulus and eCG × BCS. Cows in the CG group were less likely (28.9% vs. 33.8%; P < 0.05) to become pregnant compared with those in the EG group (odds ratio [OR] = 0.28). There were no differences in P/AI between CE and EE cows (30.9% vs. 29.1%; OR = 0.85; P = 0.56), respectively. Thinner cows not receiving eCG had lower P/AI than thinner cows receiving eCG (15.2% vs. 38.0%; OR = 0.20; P < 0.01). Treatment with eCG tended to increase serum progestesterone concentrations during the diestrus following synchronized ovulation (P < 0.10). However, the treatment used to induce ovulation did not affect CL volume or serum progesterone concentrations. In conclusion, both ECP and GnRH yielded comparable P/AI. However, eCG treatment at CIDR removal increased pregnancy rate in cows induced to ovulate with GnRH and in cows with lower BCS.     

Evaluating the success of sex-sorted semen in US dairy herds from on farm records

These data summarize on-farm records of dairy herds (n = 211) using sexed semen. Sexed semen was predominantly used at first and second service in virgin heifers, which is reflected in younger ages at AI and at calving. Conception rates at first service averaged 47% for Holstein heifers and 53% for Jersey heifers, which were ∼80% of that achieved with conventional semen. Analysis of inter-estrus intervals provides no evidence that cycle lengths are extended by use of sexed semen. Among singleton births, 89% were reported as female offspring and this rises to 90% for gestation lengths within a normal 265-295 d range. Age at calving appeared to interact with calf sex and semen type to influence the incidence of stillbirths. Semen type had no effect on the incidence of stillbirths among heifers delivering female calves. However, the incidence of stillbirths among heifers delivering male calves was greater for those conceived from sexed semen and was only partially explained by age at calving. Because the incidence of male calves from sexed semen is only 10%, the total incidence of stillbirths was not affected by semen type. In conclusion, failure to differentiate sexed from conventional semen in data recording and preferential bias in use of sexed semen in younger, more fertile females makes legitimate comparisons of sexed and conventional semen in the commercial setting difficult. When used in Holstein heifers, the average first service conception rate achieved with sex-sorted semen was 47%, which appeared to ∼80% of that achieved with conventional semen in the same herds. The percentage of female calves (89%) was consistent with expectations. After adjusting for age at calving, sexed semen had no affect on the total incidence of stillbirths, however the source for an apparent increased incidence of stillbirth among male calves born from X-sorted sperm populations requires further investigation.     

Transgenic chickpea seeds expressing high levels of a bean α-amylase inhibitor

We describe a robust and reproducible Agrobacterium-mediated chickpea transformation method based on kanamycin selection, and its use to introduce the bean AI1 gene into a desi type of chickpea. Bean AI1 was specifically expressed in the seeds, accumulated up to 4.2% of seed protein and was processed to low molecular weight polypeptides as occurs in bean seeds. The transgenic protein was active as an inhibitor of porcine -amylase in vitro. Transgenic chickpeas containing -AI1 strongly inhibited the development of Callosobruchus maculatus and C. chinensis (Col. : Bruchidae) in insect bioassays.     

Inhibition of renin secretion by platelet activating factor (acetylglyceryl ether phosphorylcholine) in cultured rat renal juxtaglomerular cells

Acetylglyceryl ether phosphorylcholine (AGEPC), commonly known as platelet activating factor, was found to strongly inhibit renin secretion in cultures rich in juxtaglomerular cells. This inhibitory action of AGEPC was accompanied by an enhanced calcium permeability of the cell membrane as evaluated from measurements of the uptake of 45Ca. Simultaneous addition of the calcium channel blocker verapamil abolished the effects of AGEPC on both renin secretion and calcium permeability. Furthermore, addition of AGEPC to the cell cultures led to a decrease of 32P-labeled phosphatidylinositol 4,5-bisphosphate and to an increase in 3H-labeled diacylglycerol, indicating an activation of phospholipase C by AGEPC.     

Preparation of hair for measurement of elements by inductively coupled plasma-mass spectrometry (ICP-MS)

The preparation of hair for the determination of elements is a critical component of the analysis procedure. Open-beaker, closedvessel microwave, and flowthrough microwave digestion are methods that have been used for sample preparation and are discussed. A new digestion method for use with inductively coupled plasma-mass spectrometry (ICP-MS) has been developed. The method uses 0.2 g of hair and 3 mL of concentrated nitric acid in an atmospheric pressurelow-temperature microwave digestion (APLTMD) system. This preparation method is useful in handling a large numbers of samples per day and may be adapted to hair sample weights ranging from 0.08 to 0.3 g. After digestion, samples are analyzed by ICP-MS to determine the concentration of Li, Be, B, Na, Mg, Al, P, S, K, Ca, Ti, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Ge, As, Se, Rb, Sr, Zr, Mo, Pd, Ag, Cd, Sn, Sb, I, Cs, Ba, Pt, Au, Hg, Tl, Pb, Bi, Th, and U. Benefits of the APLTMD include reduced contamination and sample handling, and increased precision, reliability, and sample throughput.     

Nascent Astrocyte Particles Differ from Lipoproteins in CSF

Abstract: Little is known about lipid transport and metabolism in the brain. As a further step toward understanding the origin and function of CNS lipoproteins, we have characterized by size and density fractionation lipoprotein particles from human CSF and primary cultures of rat astrocytes. The fractions were analyzed for esterified and free cholesterol, triglyceride, phospholipid, albumin, and apolipoproteins (apo) E, AI, AII, and J. As determined by lipid and apolipoprotein profiles, gel electrophoresis, and electron microscopy, nascent astrocyte particles contain little core lipid, are primarily discoidal in shape, and contain apoE and apoJ. In contrast, CSF lipoproteins are the size and density of plasma high-density lipoprotein, contain the core lipid, esterified cholesterol, and are spherical. CSF lipoproteins were heterogeneous in apolipoprotein content with apoE, the most abundant apolipoprotein, localized to the largest particles, apoAI and apoAII localized to progressively smaller particles, and apoJ distributed relatively evenly across particle size. There was substantial loss of protein from both CSF and astrocyte particles after density centrifugation compared with gel-filtration chromatography. The differences between lipoproteins secreted by astrocytes and present in CSF suggest that in addition to delivery of their constituents to cells, lipoprotein particles secreted within the brain by astrocytes may have the potential to participate in cholesterol clearance, developing a core of esterified cholesterol before reaching the CSF. Study of the functional properties of both astrocyte-secreted and CSF lipoproteins isolated by techniques that preserve native particle structure may also provide insight into the function of apoE in the pathophysiology of specific neurological diseases such as Alzheimer's disease.