Estrus induction in white rhinoceros (Ceratotherium simum)

The estrous cycle length in the white rhinoceros (Ceratotherium simum) is either 4 or 10 wk. The cause(s) for this variation as well as the poor fertility rate in captivity remains under debate in this species. Most captive adult white rhinoceros undergo long anovulatory periods without luteal activity which are considered a major reason for their low reproductive rate. In this study, the synthetic progestin chlormadinone acetate (CMA) was tested in combination with hCG or the GnRH analogue deslorelin for its efficiency to induce ovulation in fourteen females without luteal activity and in three, regular cycling females. HCG (N = 12), injectable GnRH analogue (N = 8) and GnRH analogue implants (N = 15) were given to induce ovulation after CMA treatment. Treatment success was determined using both transrectal ultrasonography and progesterone metabolite EIA analysis. A preovulatory sized follicle (3.5 ± 0.1 cm) or a corpus luteum (5.1 ± 0.7) was present on the ovary one day after induction in 93.1% of the treatments. Despite this high rate of ovarian response, ovulation rate differed between the study groups. The ovulation rate for hCG, injectable GnRH analogue and GnRH analogue implants was 66.7%, 62.5% and 93.3%, respectively. Ovulation rate in cyclic females treated with GnRH implants was 100% (6/6) compared with 89% (8/9) in females without luteal activity receiving the same treatment. The length of the estrous cycle when induced with hCG was 4 wk (85.7%). The estrous cycle when induced with GnRH analogue was predominantly 10 wk long. Two females without luteal activity treated with GnRH became pregnant. In conclusion, CMA in combination with GnRH analogue implants was highly effective to induce ovulation in white rhinoceroses and thus can contribute to efforts aimed at increasing natural mating and reproductive rates in the captive white rhinoceros population.     

Effect of cis-, trans-diamminedichloroplatinum(II) and DBP on human serum albumin

Both isomers of diamminedichloroplatinum(II) bind to albumin and induce the formation of the albumin dimer (MW approximately 140 kDa). The trans isomer exhibits a much greater tendency to induce a protein dimerization than the cis isomer. Under similar experimental conditions, the phosphonic derivative of diammineplatinum(II) (DBP) does not induce any dimer formation. The amount of bound complex per mol of human serum albumin (HSA, for an incubation time of 7 days) was found to be 6, 10.5 and 1 mol for cis-, trans-DDP and DBP, respectively. The relative fluorescence intensity of platinum-bound HSA decreases to about 55% for cis-DDP, 45% for trans-DDP and to 85% for DBP when compared to the complex-free protein, suggesting that the binding occurs in the proximity of the Trp214 residue. The structural studies (CD) have shown that only DDP-isomers cause the distinct modification of HSA native structure (alpha-helical content). Pt(II) complexes binding to HSA affect the affinity of HSA towards heme and bilirubin. High excess of DDP prevents the heme and bilirubin binding, while DBP affects this binding much less effectively due to the low amount of the protein-bound complex. Reactions of platinum complexes with albumin are believed to play an important role in the metabolism of this anticancer drug. The minor effect of DBP on HSA may indicate that the toxicity of the phosphonate analog is much lower than toxicities of DDP isomers, most likely due to kinetic reasons.     

Development of white spruce (Picea glauca (Moench.) Voss) somatic embryos during culture with abscisic acid and osmoticum, and their tolerance to drying and frozen storage

The limit of permeability of white spruce (Picea glauca [Moench.]Voss) somatic embryo cell walls to molecules was in the orderof 30 . Polyethylene glycols (PEGs) and dextrans of molecularweights greater than 1000 and 6000, respectively, produced anonpermeating (non-plasmolysing) water stress which improvedembryo development. Somatic embryos converted to plantlets atfrequencies of 76–84% following slow drying and storageat –20 C for 1 year, which was similar to the 77% recordedfor control somatic embryos slowly dried then germinated withoutfreezing or storage. Culture for 7–8 weeks with mediumcontaining abscisic acid, 3% sucrose, and 7.5% PEG 4000 yieldedsomatic embryos with five times the embryo storage lipid contentrecorded for zygotic embryos. During culture with PEG the moisturecontent of the somatic embryos decreased from 96% for immaturesuspension-cultured somatic embryos, to 47% for mature embryos.Somatic embryos cultured for 7–8 weeks survived rapiddrying to 5% moisture content, and converted to plantlets atfrequencies of 60–70%, but no somatic embryos survivedrapid drying when cultured for only 4 weeks; however, slow dryingdid induce desiccation tolerance in 3-week cultured somaticembryos. Abscisic acid was important to maintain embryos ina developmental state, but ABA alone did not induce desiccationtolerance. In order to induce desiccation tolerance a waterstress treatment was required. Tolerance of rapid drying coincidedwith moisture contents below 55%, which occurred after 5 weeksof culture in the presence of PEG 4000 and abscisic acid. Key words: Dextran, molecular weight, polyethylene glycol, triacylglycerol, water stress     

In vitro Mutation of Chrysanthemum (Dendranthema grandiflora Tzvelev) with Ethylmethanesulphonate (EMS) in Immature Floral Pedicels

Many new cultivars of chrysanthemum were derived from induced mutation. However, chemical mutagenic agents are not widely used due to their low penetration into plant tissues. The objective of the present study was to induce mutation in chrysanthemum using ethylmethanesulphonate (EMS) in immature floral pedicels, followed by the induction and production of adventitious buds in vitro. Preliminary studies on the sensitivity of pedicels to EMS revealed that the LD50 was close to 0.82% (v/v). Immature pedicels of chrysanthemum cv. Ingrid (dark pink color) were treated with 0.77% (0.075 M) EMS solution for 1 h and 45 min, which was followed by rinsing in water for 15 min and surface disinfection. Afterwards, they were cultivated in Murashige and Skoog medium (salts and vitamins) amended with 1 g l⁻¹ of hydrolyzed casein, 1 mg l⁻¹ 6-benzylaminopurine (BAP) and 2 mg l⁻¹ indole-3-acetic acid (IAA). A total of 910 plants were obtained from the pedicels treated with EMS and were evaluated at the flowering stage. Forty-eight mutants (5.2%) were obtained, deviating in petal color (pink-salmon, light-pink, bronze, white, yellow and salmon color). Most of them (89.6% of the total) were phenotypically uniform. The results showed the efficiency of EMS to induce in vitro mutation of chrysanthemum.     

Synthesis, characterization, and cytotoxic activity of copper(II) and platinum(II) complexes of 2-benzoylpyrrole and X-ray structure of bis[2-benzoylpyrrolato(N,O)]copper(II)

Copper(II) and platinum(II) complexes of 2-benzoylpyrrole (2-BZPH) were synthesized and characterized with IR, 1H and 13C NMR spectroscopies and coordination geometry with ligands arranged in transoid fashion. The crystal structure of [Cu(II)(2-BZP)2] was determined by X-ray diffraction. Death of complex treated Jurkat cells was measured by flow cytometry. The bis-chelate complexes [Cu(II)(2-BZP)2] and [Pt(II)(2-BZP)2] adopt square-planar coordination geometry with ligands, arranged in transoid fashion. Concentrations of 1-10 microM Platinum(II) complexes reduced cell survival from 100% to 20%, in contrast to the copper(II) complex which caused no cell death at a concentration of 10 microM. While the Pt(II) complexes may have damaged DNA to induce cell death, treatment with the Cu(II) complex did not induce Jurkat cell death.     

Trail-following behavior in the German cockroach (Dictyoptera: Blattellidae)

The trail-following behavior of the German cockroach, Blattella germanica (L.), was evaluated by comparing the distance between cockroach movement paths and preapplied "trails" of fecal extract. For each cockroach group tested (adult males, females, gravid females, and late instars), the mean perpendicular distance of the cockroach from the trail was significantly less than the distance from a control trail. The results indicated that the German cockroaches did exhibit trail-following behavior. Trail- following accuracy varied among the cockroach groups. The mean distance from the fecal trail ranged from 18.45 to 110.05 cm with adult males < or = adult females < or = late instars < gravid females. Very dilute fecal extract could still induce trail-following behavior in adult male cockroaches. A 5.6% concentration of fecal extract in methanol was able to induce trail-following behavior in 50% of the cockroaches. Although German cockroaches have demonstrated trail-following behavior, fecal trails are still not thought to be actively deposited. Rather, the passive distribution of fecal material within the home range results in the accumulation of trails along frequently traveled routes (i.e., between resources and the cockroach harborage).     

Synthesis of (E)-9-(1-pyrenyl)-4-hydroxynon-2-enal, a fluorescent probe of the (E)-4-hydroxynon-2-enal with retained biological properties

(E)-9-(1-pyrenyl)-4-hydroxynon-2-enal (FHNE), a fluorescent probe of (E)-4-hydroxynon-2-enal (HNE) is synthesised in seven steps and in 35% overall yield, starting from commercially available 1-pyrencarboxyaldehyde. When incubated with cultured HeLa cells this fluorescent probe penetrates cells and particularly concentrates in the region surrounding the nucleus. As the parent compound, HNE it is able to induce the activation of heat shock factor (HSF) and it is able to induce the binding of HSF to heat shock element (HSE).     

Association between hMLH1 hypermethylation and JC virus (JCV) infection in human colorectal cancer (CRC)

Incorporation of viral DNA may interfere with the normal sequence of human DNA bases on the genetic level or cause secondary epigenetic changes such as gene promoter methylation or histone acetylation. Colorectal cancer (CRC) is the second leading cause of cancer mortality in the USA. Chromosomal instability (CIN) was established as the key mechanism in cancer development. Later, it was found that CRC results not only from the progressive accumulation of genetic alterations but also from epigenetic changes. JC virus (JCV) is a candidate etiologic factor in sporadic CRC. It may act by stabilizing β-catenin, facilitating its entrance to the cell nucleus, initialing proliferation and cancer development. Diploid CRC cell lines transfected with JCV-containing plasmids developed CIN. This result provides direct experimental evidence for the ability of JCV T-Ag to induce CIN in the genome of colonic epithelial cells. The association of CRC hMLH1 methylation and tumor positivity for JCV was recently documented. JC virus T-Ag DNA sequences were found in 77% of CRCs and are associated with promoter methylation of multiple genes. hMLH1 was methylated in 25 out of 80 CRC patients positive for T-Ag (31%) in comparison with only one out of 11 T-Ag negative cases (9%). Thus, JCV can mediate both CIN and aberrant methylation in CRC. Like other viruses, chronic infection with JCV may induce CRC by different mechanisms which should be further investigated. Thus, gene promoter methylation induced by JCV may be an important process in CRC and the polyp-carcinoma sequence.     

Strains of Zucchini Yellow Mosaic Virus in Muskmelon (Cucumis melo L.)

According to the reaction of muskmelon line PI 414723, 22 natural isolates of Zucchini Yellow Mosaic Virus (ZYMV) were grouped into two pathotypes. When inoculated by isolates belonging to pathotype 0, most of the PI 414723 plants (over 70%) remained symptomless while few plants developed systemic chloronecrotic spotting and more rarely yellowing, stunting, mosaic and leaf deformations. When contaminated by pathotype 1 all PI 414723 plants developed systemic chloronecrotic spotting. Two variants were obtained from representative strains of these two pathotypes, able to induce yellowing, stunting, mosaic and leaf deformation on all inoculated PI 414723 plants. These variants could not be differentiated from their originating strains either by host range, serology or aphid transmission properties. Therefore they are regarded as belonging to a third group called pathotype 2. No relation was observed between the type of symptom developed on PI 414723 and the ability to induce a rapid wilting reaction of melon cv. Doublon (pathotype F).     

Synthesis of (E)-9-(1-pyrenyl)-4-hydroxynon-2-enal, a fluorescent probe of the (E)-4-hydroxynon-2-enal with retained biological properties

(E)-9-(1-pyrenyl)-4-hydroxynon-2-enal (FHNE), a fluorescent probe of (E)-4-hydroxynon-2-enal (HNE) is synthesised in seven steps and in 35% overall yield, starting from commercially available 1-pyrencarboxyaldehyde. When incubated with cultured HeLa cells this fluorescent probe penetrates cells and particularly concentrates in the region surrounding the nucleus. As the parent compound, HNE it is able to induce the activation of heat shock factor (HSF) and it is able to induce the binding of HSF to heat shock element (HSE).     

四种杀虫剂对白纹伊蚊(Aedes albopictus)细胞株(C6/36)的细胞遗传学效应

本文研究了四种杀虫剂:敌敌畏乳剂,灭害灵,甲基对硫磷,叶蝉散对白纹伊蚊细胞株（C6/36）染色体畸变和姐妹染色单体互换（SCE）的影响。结果表明:0.1％和0.01％浓度的敌敌畏,可使（SCE）频率明显增加,而染色体畸变率只在0.1％浓度有显著增加;0.5％的灭害灵,可使SCE频率明显增加,而染色体畸变增加不明显;甲基对硫磷和叶蝉散的实验结果表明,无论是染色体畸变和SCE频率均没有明显增加。     

Variable effects of goitrogens in inducing precocious metamorphosis in sea lampreys (Petromyzon marinus)

The ability of different goitrogens (anti-thyroid agents) to induce precocious metamorphosis in larval sea lampreys (Petromyzon marinus) was assessed in four separate experiments. Two of these goitrogens (propylthiouracil [PTU] and methimazole [MMI]) are inhibitors of thyroid peroxidase-catalyzed iodination, and three (potassium perchlorate [KClO(4)], potassium thiocyanate [KSCN], and sodium perchlorate [NaClO(4)]) are anionic competitors of iodide uptake. Because, theoretically, all of these goitrogens prevent thyroid hormone (TH) synthesis, we also measured their influence on serum concentrations of thyroxine and triiodothyronine. All goitrogens except PTU significantly lowered serum TH concentrations and induced metamorphosis in some larvae. The incidence of metamorphosis appeared to be correlated with these lowered TH concentrations in that KClO(4), NaClO(4), and MMI treatments resulted in the lowest serum TH concentrations and the highest incidence of metamorphosis in sea lampreys. Moreover, fewer larvae metamorphosed in the KSCN and low-KClO(4) treatment groups and their serum TH concentrations tended to be greater than the values in the aforementioned groups. MMI treatment at the concentrations used (0.087 and 0.87 mM) was toxic to 55% of the exposed sea lampreys within 6 weeks. The potassium ion administered as KCl did not alter serum TH concentrations or induce metamorphosis. On the basis of the results of these experiments, we have made the following conclusions: (i) In general, most goitrogens other than PTU can induce metamorphosis in larval sea lampreys, and this induction is coincident with a decline in serum TH concentrations. (ii) The method by which a goitrogen prevents TH synthesis is not directly relevant to the induction of metamorphosis. (iii) PTU has variable effects on TH synthesis and metamorphosis among lamprey species. (iv) Unlike in protochordates, potassium ions do not induce metamorphosis in sea lampreys and are not a factor in the stimulation of this event.     

脂肪酶产生菌的筛选及不对称水解合成S-(+)-萘普生

从１６个土样中分离获得一株脂肪酶产生菌Ｅ－５３＃,该菌可优先水解Ｓ－（＋）－萘普生甲酯为Ｓ－（＋）－萘普生,ｅｅ值可超过８７％。该菌经初步鉴定为芽孢杆菌,最佳产酶培养基为葡萄糖０．５％、蛋白胨０．５％、酵母膏０．２％。０．５％的橄榄油,能诱导脂肪酶的大量产生。     

Traditional and chemical techniques for stimulation of shorea javanica (Dipterocarpaceae) resin exudation in Sumatra

Resin tappers in Sumatra induce resin flow in Shorea javanica by first opening vertical rows of small (3 cm) holes in the tree trunk. After 6–12 mo, larger (10–15 cm) holes are opened between the rows of small holes for resin harvest. Informants reported that opium applied to trees increased resin yields. Application of 10% 2-chloroethylphosphonic acid to artificial trunk wounds ofS. javanica production trees increased resin yields by 110% relative to controls in 72-h trials. Wounds of previously untapped trees exuded no resin in response to the same treatment.     

水稻幼穗培养高效再生系统的建立

本文测试了不同基因型幼穗取材时期、消毒方式、4℃下处理时间、愈伤组织诱导、分化及生根条件等对水稻幼穗培养再生成株的影响,实验结果表明,在优化条件下,籼稻的绿苗分化率在85%以上,成苗率115%左右;粳稻的绿苗分化率在90%以上,成苗率130%左右;粳型广亲和的绿苗分化率、成苗率分别达到85%和115%以上。由此建立了一套高效、可靠、重复性好的水稻组织培养再生系统,为水稻遗传转化的顺利进行奠定了基础。     

Analysis of limb anomalies induced in vitro by vitamin A (retinol) in mice.

Forelimb buds of day 11 ICR-JCL mouse embryos were cultured on liquid medium consisting of 90% Eagle's MEM and 10% fetal calf serum. Experimental medium contained 10 iu/ml vitamin A alcohol (retinol). In controls four of five metacarpals chondrified and the epidermis began to keratinize after 3-4 days of culturing. In experimental explants many pycnotic cells were observed in the peripheral mesenchyme in the hand plate, especially in pre- and postaxial regions. Only 2-3 metacarpals chondrified, and keratinization was inhibited in the experimental explants. Uptake of labeled sulfate was suppresssed by vitamin A. Excess vitamin A is thus thought (1) to act directly on limb buds, and thereby to induce limb anomalies, (2) to induce cell death in the mesenchyme, (3) to suppres the formation of chondroitin sulfate, and (4) to inhibit keratinization.     

Assessment of the safety of the cationic arginine-rich peptides (CARPs) poly-arginine-18 (R18 and R18D) in ex vivo models of mast cell degranulation and red blood cell hemolysis

Our laboratory focuses on the development of novel neuroprotective cationic peptides, such poly-arginine-18 (R18: 18-mer of l-arginine; net charge +18) and its d-enantiomer R18D in stroke and other brain injuries. In the clinical development of R18/R18D, their cationic property raises potential safety concerns on their non-specific effects to induce mast cell degranulation and hemolysis. To address this, we first utilised primary human cultured mast cells (HCMCs) to examine anaphylactoid effects. We also included as controls, the well-characterised neuroprotective TAT-NR2B9c peptide and the widely used heparin reversal peptide, protamine. Degranulation assay based on β-hexosaminidase release demonstrated that R18 and R18D did not induce significant mast cell degranulation in both untreated (naïve) and IgE-sensitised HCMCs in a dose-response study to a maximum peptide concentration of 16 μM. Similarly, TAT-NR2B9c and protamine did not induce significant mast cell degranulation. To examine hemolytic effects, red blood cells (RBCs), were incubated with the peptides at a concentration range of 1–16 μM in the absence or presence of 2% plasma. Measurement of hemoglobin absorbance revealed that only R18 induced a modest, but significant degree of hemolysis at the 16 μM concentration, and only in the absence of plasma. This study addressed the potential safety concern of the application of the cationic neuroprotective peptides, especially, R18D, on anaphylactoid responses and hemolysis. The findings indicate that R18, R18D, TAT-NR2B9c and protamine are unlikely to induce histamine mediated anaphylactoid reactions or RBC hemolysis when administered intravenously to patients.     

Effect of poly(ethylene glycol) on the Ca2+-induced fusion of didodecyl phosphate vesicles

This paper reports a study of the effect of the dehydrating agent poly(ethylene glycol) (PEG) on didodecyl phosphate (DDP) bilayers and on the fusion activity of DDP vesicles as a function of the molecular weight of PEG. PEG 8K in a concentration of 10 wt % does not induce fusion. However, Ca2+-induced fusion is promoted as reflected by a lowering of the Ca2+ threshold concentration. This effect can most likely be attributed to the dehydrating capacity of the polymer. Interestingly, low concentrations (0.1 wt %) of PEG 20 K induce a moderate fusion capacity. At higher concentrations (0.5 wt %) fusion is inhibited, irrespective of the presence of Ca2+. These molecular weight dependent effects can be rationalized by taking into account that the clouding temperature differs for PEGs of different molecular weights. In the case of PEG 20K a microscopic phase separation will occur at the bilayer-water interface because PEG-PEG interactions and presumably PEG-DDP interactions are favored over PEG-water interactions. As a consequence, the DDP vesicle surface becomes covered with PEG 20K, resulting in a steric stabilization of the vesicles. This will impede or prevent, depending on the polymer concentration, the vesicles from approaching each other sufficiently close for fusion to occur.     

Cytokines modulate integrin alpha(v)beta(3)-mediated human endothelial cell adhesion and calcium signaling.

Angiogenesis is a complex process regulated by the interactions of endothelial cells with cytokines and the adhesive protein matrix. The cytokines basic fibroblast growth factor (bFGF) and tumor necrosis factor-alpha (TNF-alpha) are two of the modulators of angiogenesis. One mechanism by which these cytokines induce their effects may be through the regulation of integrin adhesion receptor activity, in particular, alpha(v)beta(3). In this study, we examined the ability of these angiogenic factors to modulate the adhesion of human umbilical vein endothelial cells (HUVECs) to immobilized disintegrins (i.e., rhodostomin and arietin), which are specific in antagonizing integrin alpha(v)beta(3) in cells. As these disintegrins were immobilized as substrates, they acted as agonists to induce HUVEC adhesion in a dose- and alpha(v)beta(3)-dependent manner. In addition, adhesion also triggered a sustained increase of intracellular free calcium. Furthermore, bFGF-primed HUVECs potentiated, but TNF-alpha primed cells attenuated, about 50% adhesion events and calcium signaling triggered by immobilized disintegrin compared to naive cells, respectively. The mechanisms of modulating alpha(v)beta(3)-dependent HUVEC adhesion by cytokines may be related to changes of integrin alpha(v)beta(3) conformation, as demonstrating the antagonistic effect of Mn(2+) on decreased adhesion by TNF-alpha pretreatment, and confirmed with flow cytometric analysis probed by anti-LIBS1 mAb. However, cytokine pretreatment did not alter the expression of this integrin on the cell surface, as determined by flow cytometry. Phosphoinositide-3 kinase may be one of the signaling molecules involved in the enhanced adhesion of bFGF-primed cells.     

激光诱发家蚕孤雌生殖的研究

采用 CO2激光扩束辐照诱发家蚕孤雌生殖,获得着色卵率约60%,转青卵率约31%,孵化率只占转青率的1.85%,全是雌蚕,其发育机制属非减数分裂型,没有出现畸形蚕,茧质未下降。从激光诱发的孤雌生殖的后代中,初步选出两个新类型的可作育种材料。 Abstrect:In order to induce silkworm(Bombyx mori)parthenogenesis,we irradiated eggs by Co2 laser.The results showed,60% colored eggs,31% average bluish eggs,only 1.85% hatchavility among the bluish eggs,all female from those eggs and so an ameiosis parthenogenesis;no malformed worm,thus similar constitution and quality to their parents;and finally two new strains which can be used to breed new strains and furhther studies.