Efficient isolation of X chromosome-specific single-copy probes from a cosmid library of a human X/hamster hybrid-cell line: mapping of new probes close to the locus for X-linked mental retardation.

We isolated X-chromosomal DNA probes from a cosmid library constructed from a single human X/hamster hybrid-cell line (C12D). One hundred human clones were isolated and used to construct a pool of X-chromosomal DNA. This DNA was digested into 0.15-2-kb fragments and subcloned into plasmids allowing the rapid characterization of new single-copy probes. These were regionally mapped and used for the detection of restriction-site polymorphisms. Together with a series of subcloned probes from individually isolated cosmids, we found seven polymorphic probes among 53 tested. Thirty-one of the probes were physically localized to different regions of the X chromosome. Four polymorphic probes map to Xq27-Xq28: DXS102 (cX38.1), DXS105(cX55.7), DXS107(cpX234), and DXS134(cpX67). These were genetically mapped by multipoint analysis relative to previously characterized loci, a mapping that resulted in the following order: DXYS1, DXS107, DXS51/DXS102, F9, DXS105, Fra-X, F8/DXS52, DXS15, DXS134. The mapping of DXS105 between F9 and Fra-X makes this probe useful for Fra-X analysis. For the linkage between FraX and DXS105, a maximum lod score of 5.01 at 4 cMorgans has been obtained in one large Dutch pedigree.     

Dominant expression of an amino-acid uptake deficiency in carrot somatic fusion hybrids

Somatic hybrids were selected previously by their ability to grow in medium containing normally inhibitory levels of the two amino acid analogs aminoethylcysteine (AEC) and 5-methyltryptophan (5MT) following fusion of protoplasts from a cell strain resistant to AEC with protoplasts resistant to 5MT. The hybrid nature of the selected clones was shown by several criteria including the presence of another resistance, azetidine-2-carboxylate (A2C), carried by one of the parental strains which was not selected for in the initial hybrid selection scheme. The characterization presented here shows that the AEC resistance in the parental strain, as well as the two somatic hybrids, was due to decreased AEC uptake. Also the 5MT resistance in the hybrids, as in the parent was caused by a feedback altered form of the tryptophan biosynthetic control enzyme, anthranilate synthase which leads to increases in free tryptophan. The A2C resistance was caused by the accumulation of free proline by a mechanism which has not been studied. These studies confirm that AEC resistance caused by decreased uptake can be expressed dominantly in protoplast fusion hybrids.Abbreviations A2C Azetidine-2-carboxylate - AEC Aminoethylcysteine - 5MT 5-methyltryptophan     

Influence of Nitrogen Fertilizers on Growth, Spore Production and Germination, and Biocontrol Potential of Trichoderma and Gliocladium

The mycelial weight of eight out of nine isolates of Trichoderma spp. and Gliocladium virens increased in media supplemented with 2000 mg/l of nitrogen (N) from the fertilizers NH₄Cl, NaNO₃, and a commercial 20–20–20. In general, the greatest increase in growth (up to 311 %) occurred with 20–20–20. The extent of growth was similar with either NH₄Cl or NaNO₃, but was less than that with 20–20–20. Measured by radial development on agar surfacesgrowth of isolates was either not affected or was constricted by supplemental fertilizers. Production of conidia by six out of eight isolates was stimulated by 20–20–20 but not by NH₄Cl or NaNO₃. Germination of conidia of all isolates, generally was high (> 85 %) on amended and nonamended agar. Chlamydospore formation by three Trichoderma isolates in liquid media was not affected by fertilizers. Antagonism or overgrowth of the pathogen Rhizoctonia solani by Trichoderma isolates in culture was reduced appreciably by NaNO₃, but was not affected by NH₄Cl or 20–20–20. Addition of 20–20–20 to natural soil did not reduce further the survival of R. solani caused by germling preparations of six out of seven Trichoderma isolates. However, reduction in survival of the pathogen caused by a T. hamatum isolate was stimulated further (45 %) by the fertilizer.     

Survival and transport of bacteria in egg washwater.

An evaluation of methods for monitoring the quality of water used to wash eggs at grading stations was undertaken to improve maintenance of bacterial viability during overnight sample transport. Bacterial content of samples at analysis would then better reflect conditions at the time eggs were washed. The interactive effects of temperature and the highly alkaline water conditions upon viability were the subjects of this study. Nine transport methods were examined for their efficacy in recovering total and coliform bacteria from recycled water used to wash eggs, and these were compared with samples analyzed at two commercial egg grading stations. Samples were shipped under test to the laboratory for analysis the following day. The survival of Staphylococcus aureus and Escherichia coli was also examined, but in a synthetic washwater matrix under various combinations of temperature (6 to 32 degrees C) and pH (9.5 to 10.5) to determine whether there was likely to be a different response to variations in transport treatment among gram-positive and -negative bacteria. S. aureus was much more resistant to the lethal effects of high pH and moderate temperature than E. coli. These results indicated that samples of high pH should be held (transported) at less than or equal to 13 degrees C to optimize bacterial survival. Considering cost, ease of manipulation, and the ability to protect both coliforms and the bacterial population as a whole, the method of choice for transport of industrial samples was the direct addition of washwater to containers in which powdered KH2PO4 and Na2S2O3 had been placed to yield final concentrations, when dissolved, of 0.2 and 0.05% (wt/vol), respectively.     

Flight motor patterns recorded in surgically isolated sections of the ventral nerve cord ofLocusta migratoria

Summary In the locust,Locusta migratoria, the pairs of connectives between the three thoracic ganglia and in the neck were transected in all possible combinations. Each of these preparations was tested for the production of rhythmic flight motor activity, with sensory input from the wing receptors intact and after deafferentation. The motor activity elicited in these preparations was characterized by intracellular recordings from motoneurons and electromyographic analyses.The motor patterns observed in locusts with either the neck or the pro-mesothoracic connectives severed (Figs. 2, 3, and 4) were very similar to the flight motor pattern produced by animals with intact connectives. The activity recorded in mesothoracic flight motoneurons of locusts with either only the meso-metathoracic connectives cut or both the meso-metathoracic and the neck connectives transected were similar to each other. Rhythmic motor activity could be observed in these preparations only as long as sensory feedback from the wing receptors was intact. These patterns were significantly different from the intact motor pattern (Figs. 5, 6, and 7). Similar results were obtained when the mesothoracic ganglion was isolated from the other two thoracic ganglia, although the oscillations produced under these conditions were weak (Fig. 8 upper). In the isolated metathorax no rhythmic flight motor activity could be recorded (Fig. 8 lower), even when wing afferents were intact.Considering the differences between the motor patterns observed in the various preparations these results suggest that the ganglia of the locust ventral nerve cord do not contain segmental, homologous flight oscillators which are coupled to produce the intact flight rhythm. Instead they support the idea that the functional flight oscillator network is distributed throughout the thoracic ganglia (Robertson and Pearson 1984). The results also provide further evidence that sensory feedback from the wing sense organs is necessary for establishing the correct motor pattern in the intact animal (Wendler 1974, 1983; Pearson 1985; Wolf and Pearson 1987 a).Abbreviations CPG central pattern generator - EMG electromyogram     

中国长臂猿的分布(英文)

中国南部分布着3种长臂猿,它们是白眉长臂猿(Hylobates hoolock)、白掌长臂猿(H.lar)和黑冠长臂猿(H.concolor)。黑冠长臂猿在我国境内有3个亚种:指名亚种(H.c.concolor),白颊亚种(H.c.leucogenys)和海南亚种(H.c.hainanus)。这些种类目前只分布在云南和广东的海南岛,但在五十年代初也生活在广西的西南部。这些地区是长臂猿属分布区的东北边界。 同它们在中南半岛的地理特征相一致,中国的3种长臂猿的分布区不相重迭,以两条大江为界。白眉长臂猿栖居在云南怒江以西,白掌长臂猿生活在怒江与澜沧江之间地带,黑冠长臂猿占据澜沧江以东。但是有一处,即黑冠长臂猿的云南保山瓦窑分布点例外,该地的标本收藏在动物研究所。 对于黑冠长臂猿海南亚种的文献记述存在不确切之处。现有较多的标本表明,海南亚种的雌性个体头冠部具有界限明显的黑色大斑块;成年雌性个体背毛呈浅棕灰色或较鲜亮的赭黄色;背中部毛较长,在40—75毫米之间。 我们所研究的标本收藏在以下单位:纽约美国自然历史博物馆,北京自然博物馆,芝加哥福地自然历史博物馆,复旦大学生物系,中国科学院动物研究所,广州华南濒危动物研究所,上海自然博物馆,上海动物园(活体),中山大学生物系。     

The effect of alkaline borohydride treatment onN-linked carbohydrates of glycoproteins

The effects of treatments of the glycoprotein ribonuclease-B, the proteins ribonuclease-A and myoglobin, and the glyco-amino acid GlcNAc(1-N) Asn with alkalil alkaline sodium borohydride, and aqueous sodium borohydride were systematically studied as a function of the concentration of the reagents, the temperature, and the length of the treatment. High-field¹H-NMR spectroscopy, chromatographic methods and amino-acid analysis were used to characterize products of the treatments of the various compounds. Our results indicate that mild alkaline borohydride treatment, as well as aqueous borohydride treatment alone, is capable of extensively degrading polypeptides and of partially releasing theN-linked glycans from ribonuclease-B. Initially, glycopeptides are produced, the peptide portion of which consists of several amino acids, which are further hydrolyzed to yield a mixture of glyco-asparagines and oligosaccharide-alditols in the ratio of 4:1. Strong alkaline borohydride treatment of ribonuclease-B is capable of completely releasing theN-linked carbohydrates as oligosaccharide-alditols.Abbreviation RNase ribonuclease     

Vaccination againstBoophilus microplus: Localization of antigens on tick gut cells and their interaction with the host immune system

Cattle have been vaccinated againstBoophilus microplus with antigens derived from partially fed female ticks. The immune response of the host lyses the gut cells of adult ticks, causing a reduction in the number, weight and reproductive capacity of engorging ticks. This response is different from the immunity that cattle acquire after repeated tick infestation. Evidence is presented that the antigens used in vaccination are located on the plasma membrane of the gut cells and it is unlikely that these antigens are secreted into the host during feeding. Vaccination using such concealed antigens may not encounter the mechanisms of immune evasion that parasites usually demonstrate.In-vitro assays suggest that vaccination immunity is not dependent on the need to stimulate cell-mediated responses. Immunoglobulin G alone, or with the aid of complement, is enough to damage tick gut.The normal function of the one protein antigen isolated so far is unknown but we speculate that it serves some vital function on the cell plasma membrane.