Symptom-modulating properties of peanut stunt virus satellite RNA sequence variants.

The symptom-modulating properties of three peanut stunt virus (PSV) satellite RNA (satRNA) sequence variants were studied. The (V)-satRNA did not affect symptom development in tobacco plants infected with PSV. The (G)- or (WC)-satRNA, on the other hand, attenuated the symptoms. In these plants, the symptoms of PSV were restricted primarily to the inoculated leaves, and in some cases, a few leaves above the inoculated leaf showed small chlorotic areas. Northern blot analysis of total nucleic acids from PSV-infected plants containing the (V)-satRNA revealed the presence of both satellite and viral RNAs in inoculated leaves as well as in systemically infected leaves. On the other hand, satellite and viral RNAs were detected in the inoculated but not in the noninoculated leaves from infected plants containing either (G)- or (WC)-satRNA. Although a decrease in the quantities of genomic RNAs 1, 2, and 3 was characteristic of all satRNA-containing plants, this effect was more evident in the case of (G)- and (WC)-satRNAs. The complete nucleotide sequences of the three satRNAs were determined and compared to the published sequence of PSV satRNA. The (V)-satRNA differed from the published sequence at two positions, whereas the (G)- and (WC)-satRNAs differed at six and eight positions, respectively. Comparison of the nucleotide sequence of the satRNA having no effect on PSV-induced symptoms with those reducing virus symptoms suggests that a single nucleotide change or as many as five nucleotide changes may distinguish between attenuating and nonattenuating satRNAs.

Contemporary issues in the treatment of alcohol dependence.

Alcoholism treatment has certainly come of age since its inception with the founding of Alcoholics Anonymous in 1935. Although AA is a "fellowship" and is not considered "treatment" per se, the growth and proliferation of professional treatment programs has spawned a significant new sector in the health care industry, with a corresponding rise in health care cost. Because the social cost due to the destructiveness of alcoholism is enormous, however, treatment appears to be a cost beneficial investment for society, both in terms of dollars expended and in terms of the lessening of human misery. Inpatient and outpatient programs appear to be effective, but simplistic comparisons are to be avoided because their populations are frequently much different, with patients with poor prognosis indicators showing up more frequently in inpatient settings. Although outpatient treatment is undeniably cheaper initially, appreciable long-term savings may not be realized because of the greater tendency for recidivism in outpatient-treated groups. Taken as a whole, studies of treatment outcome and cost effectiveness support the view that contemporary alcoholism treatment should provide multiple levels of care, with appropriate matching of patients, depending on severity and various demographic factors. This matching may be difficult to realize in actual practice because of the wide variability among alcoholic patients. Although some patient-treatment matching criteria are now in use by insurance carriers and treatment providers, the long-term efficacy of these criteria have not been tested adequately. In the end, it may evolve that the best determinant of assignment to a particular treatment level or modality might best be done by a responsible clinician exercising good judgment based on experience and training. Appropriate legal and financial accountability for this decision should be based on standards of reasonableness, consistent with criteria used in the treatment of other medical and surgical conditions. The alcoholism clinician is performing a valuable service to humanity and to society. Social policies should support this effort by making available appropriate facilities and programs for treatment, and by promoting access through destigmatization and favorable payment and reimbursement systems.

A hospital day care program for alcoholics.

A high drop-out rate for conventional outpatient counseling of alcoholics led to implementation of a hospital-based intensive day care program in which patients attend treatment activities at the hospital throughout the day for ten days. During the day care program, patients are introduced to aftercare with supportive counseling and Alcoholics Anonymous. Day care has been found to enhance program flexibility to meet specialized patient needs, and to dramatically reduce the number of drop-outs. Significant short term improvement is noted in 41.6% of patients treated with intensive day care. Intensive hospital-based day care is proving to be a low cost, effective treatment modality. Long term outcome studies are suggested.

Resistance to Bean pod mottle virus in Transgenic Soybean Lines Expressing the Capsid Polyprotein.

ABSTRACT Transgenic fertile soybean plants were generated from somatic embryos of soybean (Glycine max) cv. Jack transformed via particle bombardment with the capsid polyprotein (pCP) gene of Bean pod mottle virus(BPMV). The plant transformation vector (pHIG/BPMV-pCP) utilized in these experiments contained the BPMV-pCP coding sequence, an intron-containing GUS gene, and the hygromycin phosphotransferase gene. Southern blot hybridization analysis showed that 19 transgenic soybean plants selected for resistance to hygromycin contained the genes for GUS and BPMV-pCP. The progeny of five of these transgenic soybean plants (plants 137, 139, 157, 183, and 186) were characterized in detail. An additional transgenic plant (plant 200) contained the intron-GUS and hygromycin resistance genes, but lacked the BPMV-pCP gene and was used as a negative control. Southern blot hybridization analysis of the five transgenic plants showed the presence of three copies of the T-DNA in a similar banding pattern suggesting that they were derived from a single transformation event. Western and northern blot analyses showed that the expression levels of BPMV-pCP and pCP transcript were high in these five pCP plants. Infectivity assays with detached leaves demonstrated that all five pCP plants exhibited resistance to virus infection because they accumulated lower levels of BPMV compared with plant 200 and nontransformed controls. Unlike the T(2) progeny of line 183-1 that segregated with respect to the pCP gene and, consequently, to BPMV resistance, the T(2) progeny of the homozygous line 183-2 showed little or no symptoms in response to rub-inoculation with virions of a severe strain of BPMV. Although BPMV accumulation was evident in leaves on which viruliferous beetles were allowed a 72-h inoculation access period, the upper noninoculated leaves of the T(2) progeny of line 183-2 plants were symptomless and accumulated little or no virus. Because the progeny of this homozygous transgenic line exhibited systemic resistance, they could potentially be useful in generating commercial cultivars resistant to BPMV.

Factors affecting soybean cotyledonary node transformation.

Cotyledonary node transformation efficiency was evaluated using a sonication assisted Agrobacterium-mediated transformation (SAAT) protocol, three dissimilar A. tumefaciens strains, and explants derived from 28 diverse cultivars and/or genotypes of soybean [Glycine max (L.) Merr.]. The explants were evaluated at 10 and 45 days after co-cultivation for transformation with a binary vector containing both a GUS-intron gene and an NPTII selectable marker. The best overall strain of A. tumefaciens was determined to be KYRT1 based on stable GUS transformation of soybean cotyledonary node explants measured at the terminal 45 day evaluation point. SAAT did not increase stable transformation at 45 days post co-cultivation. SAAT was determined to significantly decrease shoot proliferation of some genotypes, but it is unclear what effect this may have on the recovery of transformed shoots. Significant differences were also detected between genotypes for transformation and shoot proliferation frequency.

Development of a binary vector system for plant transformation based on the supervirulent Agrobacterium tumefaciens strain Chry5.

This report describes the disarming of Agrobacterium tumefaciens Chry5, a strain highly tumorigenic on soybean. Disarming was achieved by removing an approximately 16.5-kb segment of the 285-kb Ti plasmid pTiChry5, including approximately 4 kb of the oncogenic T-DNA and an extended region right of the T-DNA, and replacing it with a gene for carbenicillin resistance, through homologous recombination. The deletion was confirmed with Southern analysis, and the loss of tumorigenicity was verified in tobacco and tomato plant stem inoculation assays. The deletion mutant, named KYRT1, successfully transferred the ß-glucuronidase (GUS) gene into tobacco leaf tissue, producing GUS-expressing callus which could be regenerated into viable plants. In a comparative study, the transformation efficiency of A. tumefaciens KYRT1, GV3850, and EHA105 was assayed by inoculating cotyledonary node explants. The results of this study revealed that, in a binary vector system, KYRT1 is equally or more effective than EHA105 or GV3850 at delivering DNA into soybean.

Agrobacterium tumefaciens- mediated transformation of soybean [Glycine max (L.) Merrill.] using immature zygotic cotyledon explants.

AGROBACTERIUM TUMEFACIENS: -mediated transformation of soybean [Glycine max (L.) Merrill. cv. Jack] using immature zygotic cotyledons was investigated to identify important factors that affected transformation efficiency and resulted in the production of transgenic soybean somatic embryos. The factors evaluated were initial immature zygotic cotyledon size, Agrobacterium concentration during inoculation and co-culture and the selection regime. Our results showed that 8- to 10-mm zygotic cotyledons exhibited a higher transformation rate, as indicated by transient GUS gene expression, whereas the smaller zygotic cotyledons, at less than 5 mm, died shortly after co-cultivation. However, the smaller zygotic cotyledon explants were found to have a higher embryogenic potential. Analysis of Agrobacterium and immature cotyledon explant interactions involved two Agrobacterium concentrations for the inoculation phase and three co-culture regimes. No differences in explant survival or somatic embyogenic potential were observed between the two Agrobacterium concentrations tested. Analysis of co-culture regimes revealed that the shorter co-culture times resulted in higher explant survival and higher somatic embryo production on the explants, whereas the co-culture time of 4 days severely reduced survival of the cotyledon explants and lowered their embryogenic potential. Analysis of selection regimes revealed that direct placement of cotyledon explants on hygromycin 25 mg/l was detrimental to explant survival, whereas 10 mg/l gave continued growth and subsequent somatic embryo development and plant regeneration. The overall transformation frequency in these experiments, from initial explant to whole plant, was 0.03 %. Three fertile soybean plants were obtained during the course of these experiments. Enzymatic GUS assays and Southern blot hybridizations confirmed the integration of T-DNA and expression of the GUS-intron gene in the three primary transformants. Analysis of 48 progeny revealed that three copies of the transgene were inherited as a single Mendelian locus.

New hope for impaired physicians: helping the physician while protecting patients.

The medical profession has developed a greater awareness of the problem of substance abuse among physicians, and has responded by setting programs to confront impaired physicians about their problem so that they can overcome their resistance to treatment. Nonetheless, the most effective strategy is prevention: instilling physicians-in-training with a more cautious attitude towards drug and alcohol use.

Dental management of the recovered chemically dependent patient.

People who are recovered from chemical dependency are frequently seen in dental practice. They have special needs, especially in pain management. Understanding chemical dependency and establishing a good pretreatment rapport with the patient will assist the dentist in reducing the need for postoperative analgesics. The practitioner should rely highly on nonsteroidal anti-inflammatory agents as primary analgesics for these patients. Opiate derivatives, tranquilizers, sedatives, cough syrups, and general anesthetics should be avoided.

Treatment of alcoholism: the role of the primary care physician.

The primary care physician plays an important role in early identification of the alcoholic, who may seek treatment for a problem other than alcoholism. Confrontation begins with a complete history and ends with an invitation to the patient to participate in definitive treatment. During detoxification the patient can be helped to establish a long-term relationship with the physician and with Alcoholics Anonymous.

Propoxyphene dependence: an update.

Despite the initial claims that propoxyphene (Darvon) provides analgesia without risk of dependence, there is growing international recognition of its addictive potential, with documentation of both psychological dependence and physical withdrawal symptoms. However, many physicians remain uninformed of the true nature of the drug and its potential for abuse and addiction. In terms of frequency of addiction among users, propoxyphene may be less dangerous than most narcotics, but it is more available by prescription and less expensive then illegal opiates, making it a prime target for both abuse and addiction. At present, increased caution by prescribing physicians is indicated. Patients now taking the drug should be carefully monitored for signs of abuse or dependence.

Fatty acid alteration by a delta 9 desaturase in transgenic tobacco tissue.

Nicotiana tabacum tissue was transformed with a rat stearyl-CoA desaturase gene. Gas chromatographic analysis showed an increase in monounsaturated 16 and 18 carbon fatty acids in selected transformed calli and leaves. Fractionation of lipid classes indicated that palmitoleic acid was found in the phosphatidylcholine fraction of desaturase-transformed leaves, but not in leaves transformed with vector sequences. Plant transformation was verified by polymerase chain reaction (PCR) amplification of total leaf DNA.

A comparison of free-hand vs laser-guided long-axis ultrasound techniques in novice users.

OBJECTIVE: The increasing use of point-of-care ultrasonography for targeted procedures justifies a device that helps both novices in training and experts perform the long-axis needle approach. The initial success of traditional needle guidance devices in reducing the time of target procedures is not universal and they can be cumbersome. We aim to investigate whether the less bulky and previously untested laser guide can succeed in reducing procedure time in novice ultrasonographers. METHODS: 82 medical students with no ultrasound experience volunteered. Random allocation determined whether, during a targeted procedure in a turkey breast and olive phantom, participants were assisted by the laser guide or not. The time taken to pierce the target was recorded at 1-cm depth. RESULTS: The mean procedure time in the laser-assisted (LA) group was 25.1 s (14.0 s; 18.0-25.0 s). The mean procedure time in the free-hand group was 45.5 s (23.0 s; 7.0-55.0 s). The procedure time in the LA group was significantly reduced (p<0.01). CONCLUSION: The laser guide significantly improved procedure times. It is felt that the cheaper, smaller, easy to integrate, sterile and more user-friendly laser guidance unit may be a better alternative to the needle guide in improving procedure times for the novice ultrasonographer or to assist the expert, during training for, or performance of, ultrasound-guided targeted procedures. ADVANCES IN KNOWLEDGE: Following from the prototype paper, this is the first study to investigate the effectiveness of attaching a laser-guidance device to an ultrasound probe. The device succeeded in reducing the procedure times of targeted procedures.

Maternal haploids of Petunia axillaris (Lam.) B.S.P. via culture of placenta attached ovules.

Hybridization of Petunia axillaris and P. parodii with Nicotiana tabacum was attempted using the method of in vitro pollination and fertilization. Seedlings were produced when the Petunia species and N. tabacum were used as the maternal parents; however, most of these had the identical somatic chromosome complement of the maternal parent. With crosses involving P. axillaris as the maternal parent, a low frequency of haploids was also produced. Due to the potential of haploids in basic and applied genetic research, additional experiments were carried out to determine whether in vitro pollination was necessary to stimulate haploid production and to more closely define the optimal time for ovule excision and culture. Four treatments were applied to accomplish these objectives. They were: placentas cultured prior to the time of anthesis, with and without pollination, and placentas cultured after the time of anthesis, with and without pollination. In vitro pollination had no effect on the frequency of haploids produced. Placenta attached ovules cultured prior to the time of anthesis produced significantly more haploids than those cultured after anthesis. The preanthesis treatment produced a frequency of 6.5 haploids per 100 ovaries cultured. The culture of placenta attached ovules provides an alternative to anther culture as a means for haploid production.

Genetic parameters in aKy 14 xKy Ex 42 burley population ofNicotiana tabacum L.

In the F2 generation of the cross between burley varieties,Ky 14 andKy Ex 42, estimates of additive genetic variance were significant for yield, days to flower, and number of leaves. All estimates of dominance variance, additive X additive variance, and interactions between genetic variances and years were smaller than twice their standard errors. The gain expected from selecting and recombining the best 10% of theF 2 plants ranged from 4% for number of leaves to 8.6% for yield. Additive genetic correlations between pairs of agronomic characters were positive. Yield and number of leaves were negatively correlated with total alkaloids.

Using the resources of AA in treating alcoholics in a general hospital.

At Cleveland Metropolitan General Hospital a small number of staff and a large number of volunteers from Alcoholics Anonymous have collaborated to meet the treatment needs of the community's alcoholics. Recovered alcoholics serve as volunteers staffing a friendship room, located close to the emergency room and open weeknights between 6 and 10 p.m. They also provide counseling on AA, escort patients to AA meetings, help in postdischarge follow-up, and act as casefinders in the community. The hospital staff have become more aware of problems of alcoholism and are better able to treat alcoholics and to provide counseling and referral services.

Identification of aneuploids in Nicotiana tabacum by isozyme banding patterns.

A biochemical system was devised to identify aneuploids of Nicotiana tabacum. Leaf tissue from 6 nullihaploids, 4 nullisomics, and 10 monosomics was analyzed electrophoretically on slab acrylamide gels. The staining systems used were for peroxidase, esterase, superoxide dismutase, malate dehydrogenase, and glutamate oxaloacetate transaminase. Nullihaploids and nullisomics could be distinguished from each other and from haploid or disomic types by their unique isozyme banding patterns. The banding patterns of the monosomics closely resembled those of the disomic. Morphologically similar aneuploids from different populations had similar isozyme banding patterns.