Postnatal changes in enzyme activities of rat myocardial adenine nucleotide catabolic pathway

Three catabolic enzymes, 5'-nucleotidase (5'NT), adenosine deaminase (ADA), purine nucleoside phosphorylase (PNP) and one anabolic enzyme, myokinase (MK) involved in adenine nucleotide (AN) metabolism were studied in myocardium from 4 to 105 day old rats. The specific enzyme activities (nmoles/min/mg protein) at day 4 were 35.3 for 5'NT, 28.4 for ADA, 43.3 for PNP, and 5 X 10(3) for MK. At day 7, 5'NT, activities rose to 450%; PNP and ADA 150%; and MK 120%; of the day 4 level. The activities of the three catabolic enzymes were elevated for one or two weeks then declined rapidly. By day 34, they were slightly above the adult values. MK activity displayed a different time course. It continued to increase slowly with age after the initial surge. Compared to the adult heart, the total activities of these catabolic enzymes in the one- to three-week-old heart were 30% to 220% higher. This transient elevation in AN catabolic enzyme activities may be related to active DNA synthesis and cell proliferation occurred in the rat myocardium during the same period.     

Association of Epstein-Barr virus early antigen diffuse component and virus-specified DNA polymerase activity.

The role of Epstein-Barr virus (EBV) early antigen diffuse component (EA-D) and its relationship with EBV DNA polymerase in EBV genome-carrying cells are unclear, EBV-specified DNA polymerase was purified in a sequential manner from Raji cells treated with phorbol-12,13-dibutyrate and n-butyrate by phosphocellulose, DEAE-cellulose, double-stranded DNA-cellulose, and blue Sepharose column chromatography. Four polypeptides with molecular masses of 110,000, 100,000, 55,000, and 49,000 daltons were found to be associated with EBV-specified DNA polymerase activity. A monoclonal antibody which could neutralize the EBV DNA polymerase activity was prepared and found to recognize 55,000- and 49,000-dalton polypeptides. An EA-D monoclonal antibody, R3 (G. R. Pearson, V. Vorman, B. Chase, T. Sculley, M. Hummel, and E. Kieff, J. Virol. 47:183-201, 1983), was also able to recognize these same two polypeptides associated with EBV DNA polymerase activity. It was concluded that EBV EA-D polypeptides, as identified by R3 monoclonal antibody, are critical components of EBV DNA polymerase.     

The transmission of an avian filarioid Cardiofilaria nilesi to the jird, Meriones unguiculatus

This paper reports the experimental transmission of a bird parasite into jirds. Infective larvae of Cardiofilaria nilesi obtained from laboratory colonized Coquillettidia crassipes mosquitoes which had fed on an infected chicken were inoculated subcutaneously into jirds. The number of larvae per jird varied from 10 to 228. Microfilaraemia appeared 22 to 89 days after inoculation of the infective larvae. Experiments were carried out with 24 jirds through six generations extending over a period of 22 months and 17 produced patent infections. At present 8 infected jirds are being maintained in the laboratory; their patent periods ranging from 6 to 13 months. However, the longest patent period observed was about thirteen months. The percentage of adults recovered in autopsied jirds ranged from 0 to 40 with an average of 16. The chicken showed a microfilarial periodicity with the peak microfilarial density around 2200 hours. However, in jirds there was a change in sub-periodicity. This model in the jird may be very useful for the screening of filaricides and in immunological work.     

Anti-histone antibodies in idiopathic and drug-induced lupus recognize distinct intrahistone regions

We have identified regions within core histones that are antigenic for autoantibodies in systemic lupus erythematosus (SLE) and drug-induced lupus. An immunoblotting technique was used to determine the reactivity of lupus antibodies for intact histones and for trypsin-resistant histone fragments that lack the amino- and carboxyl-terminal amino acids that are normally exposed in native nucleosomes. In SLE, the predominant anti-histone response was restricted to epitopes in the trypsin-sensitive regions. Of 20 SLE sera that had strong antibody activity for multiple intact histones, 17 showed minimal activity with any of the corresponding trypsin-resistant fragments. A markedly different pattern of reactivity was present in sera of patients with procainamide (Pr)-induced lupus in which antibodies to H2A, H2B, and the H2A-H2B complex had strong fragment activity. Interestingly, recognition of trypsin-resistant fragments was also noted in a small number of SLE sera that contained antibodies to the H2A-H2B complex. In contrast to both SLE and Pr-induced lupus, antibodies induced by hydralazine (Hy) reacted primarily with H3 and H4. Furthermore, these antibodies bound equally well to the corresponding trypsin-resistant regions that are thought to be relatively unexposed in native nucleosomes. Thus, the specificities of anti-histone antibodies in SLE, Pr-induced lupus, and Hy-induced lupus are markedly different, but in each disease reactivity appears to be restricted to a limited number of histone determinants. The data raise the possibility that autoantigen in the form of native nucleosomes may be recognized in SLE and possibly in Pr-induced lupus. In contrast, the propensity of Hy to induce autoantibodies to determinants usually not recognized in SLE or Pr-induced lupus may suggest a different immunogenic stimulus in this disease.     

Purification and spectroscopic properties of 124-kDa oat phytochrome

A simplified procedure for the isolation and purification of 124-kDa phytochrome from etiolated Avena seedlings has been developed using the method of ammonium sulfate back-extraction. After hydroxyapatite chromatography of seedling tissue extracts, the pooled phytochrome was subjected to ammonium sulfate back-extraction instead of the usual application to an Affi-Gel Blue column. The resulting phytochrome had specific absorbance ratios (SAR = A666/A280) ranging from 0.85 to 0.95. Subsequent Bio-Gel filtration chromatography yielded highly pure 124-kDa phytochrome with SAR values ranging from 0.99 to 1.13. The absorption maxima of 124-kDa phytochrome were at 280, 379, and 666 nm for the red absorbing form of phytochrome (Pr) and at 280, 400 and 730 nm for the far-red absorbing form (Pfr). The A730/A673 ratio in Pfr was found to be 1.5 to 1.6. The mole fraction of Pfr under red light photoequilibrium was 0.88. No dark reversion was detected within 5 h at 3 degrees C. A photoreversible far-uv-circular dichroism was observable with all phytochrome preparations examined. Fluorescence and phosphorescence lifetimes were measured to further characterize the differences between the phytochromes prepared under different conditions. The Trp fluorescence and phosphorescence lifetimes of Pr and Pfr with the chromophore "X", probably polyphenolic in nature, were significantly shorter than those of phytochrome without the contaminant X. The short lifetime of the fluorescence of the Pr chromophore is attributable to X in the former.     

Even more new taxa from South and East Anatolia II

10 new Turkish taxa are described:Arenaria eliasiana, A. sivasica, A. monscragus, A. angustifolioides; Campanula lycica; Scutellaria orientalis subsp.tortumensis; Stachys choruhensis, S. tundjeliensis; Calamintha caroli-henricana; Aristolochia rechingeriana, the latter two species named in honour ofKarl Heinz Rechinger;Allium vuralii. Dedicated to Prof. DrKarl Heinz Rechinger on the occasion of his 80th birthday. For part I see Pl. Syst. Evol.154, 111–128.     

冬小麦春化过程中可溶性蛋白质组成的变化与形态发生的关系

冬小麦“农大139”经40天左右的春化处理才能迅速而整齐地抽穗,但经14—21天低温处理,已经具有在夏季抽穗的可能性,虽然抽穗推迟且极不整齐;再将春化时间延长,则抽穗百分比增加,且从播种到抽穗的时间缩短。这表明,春化过程中低温对发育的作用有两种效应:前期低温是诱发生理状态的转变,后期低温则只具有加速发育的作用,两个时期的转变是在春化的中期。蛋白质合成抑制剂乙基硫氨酸和对-氟苯丙氨酸能抑制冬小麦的春化,抑制时期也是在春化过程的中期。不同时间低温处理后冬小麦幼芽中可溶性蛋白质含量及组成发生了变化,春化过程中期(低温处理14天之后)不仅含量比对照增加了一倍,而且有新的蛋白质谱带出现。春麦中无类似现象,未经低温处理的春麦已含有冬麦中新出现的谱带。说明冬小麦春化过程的第14—21天左右是与春化过程有关的蛋白质合成的关键时期,该时期新合成的蛋白质与植株的发育状态之间存在着密切的相关关系。     

Regeneration of leaf mesophyll protoplasts of tomato cultivars (L. esculentum): factors important for efficient protoplast culture and plant regeneration

Conditions were established for efficient plant regeneration from four freshmarket cultivars of Lycopersicon esculentum. In order to increase the yield of viable protoplasts which are able to sustain cell divisions, the donor plants are preconditioned by incubation at 25°C in the dark for 18 hours, followed by a cold treatment at 4°C in the dark for the last 6 hours, prior to protoplast isolation. Browning of the dividing cell colonies can be prevented by culturing protoplasts in 100 l droplets of low-melting agarose, surrounded by liquid medium. Alternatively, protoplasts can be cultured in liquid medium. In both procedures the plating efficiencies and percentage of shoot regeneration are increased, only when dilutions were performed with auxin-free culture medium. Shoot regeneration is obtained by using a two step procedure: initiation of greening of microcalli on a medium containing 0.2 M mannitol and 7.3 mM sucrose, which is followed by shoot development on a mannitol-free medium containing 0.5 M sucrose. In this way, plants can be regenerated within 3 months from the hybrid cultivars Bellina, Abunda, Sonatine and also from the true seedline Moneymaker. The latter one showed the highest regeneration frequency (30%).Abbreviations BAP 6-Benzylamino purine - 2,4-D 2,4-dichlorophenoxy acetic acid - IAA indole acetic acid - MES 2-(N-morpholino)- ethane sulfonic acid - NAA naphthalene acetic acid - PE plating efficiency     

Preparation of base-modified nucleosides suitable for non-radioactive label attachment and their incorporation into synthetic oligodeoxyribonucleotides.

A very mild and efficient procedure has been developed for the preparation of C-5 substituted deoxyuridines. The substituent carries a masked primary aliphatic amino group. These compounds are readily converted into their phosphoramidites and can be used to prepare oligonucleotides carrying one or more aliphatic amino groups. Fluorescein isothiocyanate coupled to these compounds gives oligonucleotide probes carrying multiple fluorescein labels. These compounds have a free 5'-hydroxy group enabling additional 5'- end radioactive labelling for evaluation of their hybridization characteristics. It was found that oligonucleotides carrying a long (11 atom) linker arm to the fluorescein hybridize more efficiently to mRNA than those carrying a short (4 atom) arm. The long linker arm derivatives are comparable to underivatized oligonucleotides in hybridizing to mRNA.     

Jejunal free autograft: analysis of complications and their resolution

Review of 101 patients who underwent 111 free jejunal autografts has demonstrated an absolute procedural failure rate of 13.5 percent. Salvage reconstruction with a second jejunum was successful in six of nine patients and one third-time jejunum was successful, giving an overall salvage rate of 70 percent. There were 33 patients experiencing pharyngocutaneous fistulas, 20 of whom had been previously irradiated. Of these patients, 15 experienced spontaneous closure and 9 others had successful surgical correction. The mortality rate was 5 percent. Eighty-three percent of patients were restored to adequate per oral alimentation. The jejunum, despite its relatively high complication rate, is an excellent method for pharyngoesophageal reconstruction, expeditiously providing return to function for patients with late-stage disease.