Firearm injury prevention counseling by pediatricians and family physicians. Practices and beliefs.

OBJECTIVES: To ascertain and compare beliefs, attitudes, and counseling practices of primary care physicians of children and adolescents regarding firearm injury prevention counseling. DESIGN: Cross-sectional survey. SETTING: State of Washington. SUBJECTS: All active members of the state chapters of the American Academy of Pediatrics and American Academy of Family Physicians. A total of 979 pediatricians and family physicians (53%) responded to the survey after two mailings. MAIN OUTCOME MEASURES: Attitudes, beliefs, and current practices with regard to firearm safety counseling among families of child and adolescent patients. RESULTS: Only 25% of pediatricians and 12% of family physicians currently counsel more than 5% of their patients. Pediatricians were more likely than family physicians (70% vs 46%, P < .001, chi 2 test) to believe that physicians have a responsibility to counsel families about firearm safety. Pediatricians recommended removing guns from the home more frequently than family physicians (32% vs 19%, P < .001, chi 2 test), but most physicians of both specialties perceived that parents are rarely receptive to this advice. However, 97% of physicians from both specialties agreed that firearms should be stored locked separately from ammunition, and a substantial majority believed that parents would be receptive to this advice. Compared with physicians who owned guns (32%), non-owners were 15 times more likely (odds ratio, 15; 95% confidence interval, 10 to 23) to agree that families with children should not keep firearms in the home. CONCLUSIONS: Few primary care physicians who see children and adolescents currently counsel families about firearm safety, although many agree that they have such a responsibility. At least half of these physicians would potentially benefit from an intervention to improve their knowledge of and counseling skills on this topic.

Transformation of maize embryogenic calli mediated by ultrasonication and regeneration of fertile transgenic plants.

An insecticidal protein gene isolated fromBacillus thuringiensis was transferred into maize by using ultrasonication. The fertile transgenic plants and their progeny were obtained. The Southern hybridization results indicated that the foreign gene had integrated into the maize genome. It has been found that the acoustic intensity and the duration of treatment are the important parameters influencing transformation efficiency by ultrasonication. The maximum relative transformation frequency of 34.1 % was achieved after 30 min of sonication at 0. 5 W/cm(2) acoustic intensity. With appropriate parameters the ultrasonication can make a number of micropores formed on the cell surface and minimize the treatment damage to the foreign DNA molecules, thus facilitating the DNA molecules to enter the cells.

[Transgenic Brassica napus resistant to turnip mosaic virus].

A system for obtaining regenerated plantlets of "double low" Brassica napus by using cotyledonary petioles as material was established. The turnip mosaic virus coat protein (TuMV-CP) gene inserted in the binary vector pBTu in the Agrobacterium tumefaciens LBA 4404 was integrated into Brassica napus through co-culture of cotyledonary petioles with LBA4404, and the material in co-culture was selected under the stress of Kanamycin. Regenerated plantlets were obtained, and the specific TuMV-CP gene was proved to be integrated into the genomic DNA of the regenerated plants by the specific PCR amplification, dot hybridization and Southern blot. All these transgenic plants were proved to be resistant to virulent TuMV by virus challenge in different degree.

cDNA cloning and sequence analysis of NIb gene of soybean mosaic virus.

cDNA of soybean mosaic virus (Beijing isolate, SMV-BJ) has been synthesized, using viral genomic RNA as a template and random hexanucleotides as primers. Based on the sequences of SMV-BJ coat protein (CP) gene as well as SMV- and WMV-II-related regions, oligonucleotides were made as primers for polymerase chain reaction (PCR). NIb gene of SMV-BJ was amplified by PCR, and cloned into pBluescript SK. The complete sequence was determined. The comparison of NIb genes between SMV-BJ and WMV-II (USA) shows that similarities for nucleotide sequence reach 80.3%; and the deduced amino acid sequence, 91.3%. In consideration of the high identities in between the CP gene and the 3'-non-coding region between them, WMV-II might be considered as a watermelon strain of SMV. Besides, some unexpected sequences were found in the 3'-region of 2 NIb gene clones. Following modification and splicing, a binary vector of NIb gene has been constructed for its expression in higher plant for the purpose of studying the possible replicase-mediated resistance in polyviruses.

[Nucleotide sequence of the toxic domain of an insecticidal protein gene from B. thuringiensis subsp. kurstaki HD-1].

Two crystal protein genes, the 5.3kb and 6.6kb class respectively, from Bacillus thuringiensis subsp. kurstaki HD-1 (B. t HD-1) had been cloned previously. Based on the classification system of Hofte and Whiteley, these two genes should belong to Cry I A (b) and Cry I A (c) gene type respectively. The nucleotide sequence of the toxic domain of this Cry I A (c) gene from B. t kurstaki HD-1 is firstly reported here and compared with that of Cry I A (c) gene from B. t HD-73 and Cry I A (b) gene from B. t HD-1.

[Purification, serology and detection of grapevine leaf roll virus].

Closterovirus-like particles associated with grapevine leaf roll disease were purified from stem phloem tissue by differential centrifugation and cesium sulphate-sucrose density gradient centrifugation. The particles were between 660-2000 nm long. Most of them were about 1400 nm long and also decorated by the antiserum against GLRV NY-1 isolate. The purified preparation was tested for their serological relatedness in indirect ELISA. The results indicated that these closterovirus-like particles reacted with serotype II, III and IV of GLRV. The antiserum to NY-1 (type III) reacted more strongly than type IV (to VA-4) and type II. (to 1800 nm). In sodium dodecyl sulfate immunodiffusion test, the extract of diseased petiole reacted with serotype III, IV and II. It is possible to be infected by 2 or 3 closterovirus-like particles in China. We have made antiserum to the purified closterovirus-like GLRV and set up a PAS-ELISA to detect GLRV-free grapevine plantelets. We have obtaimed 21 varieties of GLRV-free and GFLV-free grapevine. They shown high quality and quantity in field test.

[Construction of shuttle vector containing delta-endotoxin gene of Bacillus thuringiensis].

The Bacillus thuringiensis toxin gene CryIA(c) was inserted into the shuttle vector pBE-2 to construct pAMY for expressing the B.t. gene in both Gram-negative and -positive bacterial systems. pAMY was introduced into wild type Bacillus cereus, B.brevis and B.subtilis by electroporation. Transformants containing delta-endotoxin gene produced proteins reacted with B.t. crystal protein antibody. Upon biological toxicity tests, the transformants gave a mortality of 100% against Ostrinia furnacilis, 58.8% against Heliothis armigera and 100% against Heliothis assulta. The ability of promoting plant growth of the original strains is retained.

Application of biotechnology in plant agriculture in China.

Early in 1984, when biotechnology was implemented as one of the eight priority projects of the "Seventh Five-Year Plan," it was fully realized that China, a large crop consumer, must pay much attention to the development of agrobiotechnology. Later, in 1986, it was again listed as one of the priority projects in the "863" program, the long-term, high-technology program of China. At that time, the strongest field of biotechnology in China was the field of plant tissue culture which had been developing for over a decade. Several new cultivars of rice, wheat, and tobacco were created using anther cultures; virus-free meristem micropropagation had been applied in the control of potato degeneration and of some viral diseases of ornamental plants; regeneration of whole plants from plant protoplasts or calli has succeeded with tobacco, carrot, and canola. By contrast, the field of transgenic plants was just at its beginning, whether it could be a successfully applied technology still remained to be answered, while applications in transgenic animal research were unknown [1].

[Neonatal blennorrhea caused by Chlamydia trachomatis]. / Blennorrhoea neonatorum verursacht durch Chlamydia trachomatis.

Chlamydial conjunctivitis was diagnosed in a 5-day-old male newborn. Under treatment with erythromycin the clinical picture of intense swelling of the lid and the copious purulent discharge abated during the following 2 days. Chlamydia trachomatis has become worldwide the most prevalent causative agent of neonatal conjunctivitis.

The cloning, sequence analysis, and expression in E. coli of coat protein gene of grapevine fanleaf virus Gh.

Fragments of 5' end and 3' end of the coat protein gene of a Chinese strain grapevine fanleaf virus (GFLV-Gh) were separately amplified through RT-PCR. The two products were cloned and ligated into a complete full-length coat protein gene via an inherent BgIII restriction site. The entire gene contains 1512 nucleotides which encode a 504 amino acids coat protein, molecular weight approximately 56 kDa. Comparison with GFLV F-13 shows 88.4% homology in nucleotides and 95.8% similarity in deduced amino acid residues. Also this coat protein gene was expressed in E. coli DH 5 alpha.

[Procedures in fulminant courses of necrotizing enterocolitis]. / Vorgehen bei fulminanten Verlaufsformen der nekrotisierenden Enterocolitis.

14/42 neonates suffering from N.E.C. developed the clinical stage III (intestinal necrosis, septicemia) and/or IV (intestinal perforation, peritonitis) within 24-48 h ("fulminant" course). 2/14 deteriorated dramatically and died before operation. In 12/14 cases, the general condition could be improved and immediate laparotomy was performed (2/12 died). Management and factors affecting prognosis are discussed. Following surgery, there was no significant difference in survival rates between "fulminant" and protracted course N.E.C.

Study on replicase (subunit) gene of papaya ringspot virus cloning, sequencing and construction of higher plant expression vector.

The ds-cDNA was synthesized using genomic RNA of papaya ringspot virus (PRSV) as template. The blunt-ended cDNA was cloned into the EcoRV site of vector pBluescript SK. From the recombinants, the NIb gene of PRSV was obtained, and its complete sequence was also determined. After the NIb gene was modified through PCR, a binary vector of PRSV-NIb gene was constructed for its expression in higher plants.

[A need for surgical treatment in necrotizing enterocolitis?]. / Notwendigkeit der operativen Behandlung bei NEC?

Based on the experience with 57 neonates with NEC, the significance of diagnostic methods (especially ultrasonography, paracentesis, intraoperative photometry) for treatment and results is discussed.

Cloning of coat protein gene of soybean mosaic virus and its expression in Escherichia coli.

The 3'-terminal genomic region of the Beijing isolate of soybean mosaic virus (SMV-BJ) has been cloned through technique of polymerase chain reaction (PCR). We have analyzed the nucleotide sequence of 3' region of SMV-BJ genome. Comparisons of the nucleotide and deduced amino acid sequence of SMV-BJ coat protein gene with those of SMV-N strain show 93.4% and 98.5% identity between them, respectively. Alignments of the 3' non-coding sequence in pairs with that of SMV-N strain show homology of 88.8%. It has been found that the SMV coat protein gene is expressed in E. coli by western blot analysis. The coat protein produced in E. coli has the same electrophoretic mobility as the authentic coat protein.

A comparative study of the cowpea and bean strains of southern bean mosaic virus.

Features of the primary structure and translation of the genomic RNAs of the cowpea and bean strains of southern bean mosaic virus have been investigated in order to assess the similarity of the two viruses. The sequence of 400 bases at their 3' termini have been determined. These include the 3' noncoding regions and extend well into the coat protein cistrons. The noncoding regions (136 bases for the cowpea strain RNA and 129 bases for the bean strain RNA) show no obvious sequence homology. However, extensive base as well as amino acid sequence homology exists in the coding region. RNAs from both strains have a small protein attached to their 5' terminus-the protein in the cowpea strain being the smaller of the two. In vitro studies show that there are similarities in the overall mode of translation of the genomes of the two viruses. Although corresponding proteins are synthesized they differ in size.

Inhibition effect of transgenic tobacco plants expressing snowdrop lectin on the population development of Myzus persicae.

The cDNAs of snowdrop lectin mature protein and its precursor protein, GNA12 and GNA34, were inserted downstream of a 35S promoter with a double enhancer and a "omega" fragment of TMV-RNA cDNA in the binary vector pBin438, or the phloem-specific CoYMV promoter in the vector pBcop1, respectively, resulting in the construction of four plant expression vectors pBGna12, pBGna34, pBCGna12, and pBCGna34. Leaf stripes of Nicotiana tabacum var. K326 were transformed with A. tumefaciens LBA4404 harboring the above expression vectors, respectively. PCR and Southern blot analysis showed that foreign GNA genes were inserted into the genome of transformed tobacco plants. Western blot analysis indicated that GNA could be expressed efficiently up to 0.08-0.15% of total soluble proteins in transgenic tobacco plants with pBGna34 or pBCGna34, while in those with pBGna12 and pBCGna12 GNA were hardly detected by immunoassay. The results from insect bioassay with a peach aphid (Myzus persicae) demonstrated that the transgenic plants of pBGna34 and pBCGna34 were aphid-resistant as shown by a 45-60% reduction in insect population density, with some individual transgenic lines being reduced by over 90%. In addition, it was evident that the 35S promoter with a double enhancer and CoYMV promoter had similar abilities to direct the GNA gene to express in transgenic tobacco plants, but because the CoYMV promoter drives the foreign gene in a phloem-specific expression manner, the transgenic plants of pBCGna34 showed higher aphid resistance.

[Cloning of AHA gene from Amaranthus hypochondriacus and it's aphid inhibitory effect in transgenic tabacco plants].

Using total DNA isolated from Amaranthus hypochondriacus as template, Amaranthus hypochondriacus agglutinin AHA gene was amplified by PCR and cloned. Sequence analysis results showed that this gene is consisted of 2453 base pairs including one 1538 bp intron and two exons of 212 bp and 703 bp respectively. After inverse PCR amplification, coding region of AHA gene was obtained. AHA gene with it's intron (AHAg) and withou intron (AHAc) were inserted downstream of 35S promotor in the binary vector pBin438 resulting in the construction of two plant expression vectors pBAHAg and pBAHAc repectively. Leave explants of Nicotinana tabacum var. SR1 were transformed with A. tumefaciens LBA4404 harbouring the above expression vectors. Results from PCR and Southern blot analysis showed that AHA genes were inserted into the genome of transformed tobacco plants. Immunodot blot analysis indicated that AHA was expressed in transgenic plants. The results from insect bioassay with peach aphid (Mizus persicae) showed that the transgenic plants of pBAHAg and pBAHAc were aphid resistant, evidenced by a 57%-48% reduction in insect population density, some plants were more than 85%. The aphid resistance of transgenic plants transformed with AHAg gene as judged by aphid inhibition rate was higher than that of plants transormed with AHAc gene indicating that the intron in AHAg may be favorable for expression of AHA in transgenic plants.

Delayed seroconversion to HIV in a hemophiliac.

In a 14-year-old boy with severe hemophilia B, HIV seroconversion was observed about 2 years after substitution therapy had been changed to heat-treated PPSB concentrates. This shows that HIV-antibodies may appear in hemophiliacs after time periods longer than generally assumed.

Transfer of shuttle vectors containing Bacillus thuringiensis toxin gene into wild-type B. cereus, B. brevis and B. subtilis by electroporation.

Gram positive and negative bacterial shuttle vectors carrying Bacillus thuringiensis (B.t.) toxin gene were introduced by electroporation into wild-type Bacillus cereus, B. brevis and B. subtilis. The transformation efficiencies for these bacteria were around 10(1)-10(4) transformants per micrograms DNA based on the numbers of neomycin- and ampicillin-resistant colonies produced. The structure of the transferred plasmids proved identical with the original ones both in size and restriction pattern. Toxicity assays showed that the transformants gave a mortality of 90-100% against caterpillar of Heliothis assulta, indicating that the gene function was not changed by electroporation.