A new class of tobacco chitinases homologous to bacterial exo-chitinases displays antifungal activity

A novel chitinase gene of tobacco was isolated and characterized by DNA sequence analysis of a genomic clone and a cDNA clone. Comparative sequence analysis of both clones showed an identity of 94%. The proteins encoded by these sequences do not correspond to any of the previously characterized plant chitinases of classes I-IV and are designated as class V chitinases. Comparison of the chitinase class V peptide sequence with sequences in the Swiss Protein databank revealed significant sequence similarity with bacterial exo-chitinases from Bacillus circulans, Serratia marcescens and Streptomyces plicatus. It was demonstrated that class V chitinase gene expression is induced after treatment of tobacco with different forms of stress, like TMV-infection, ethylene treatment, wounding or ultraviolet irradiation. Two related chitinase class V proteins of 41 and 43 kDa were purified from Samsun NN tobacco leaves inoculated with tobacco mosaic virus. The proteins were purified by Chelating Superose chromatography and gel filtration. In vitro assays demonstrated that class V chitinases have endo-chitinase activity and exhibit antifungal activity toward Trichoderma viride and Alternaria radicina. In addition, it was shown that class V chitinase acts synergistically with tobacco class I beta-1,3-glucanase against Fusarium solani germlings.     

Localization of absorbers in scattering media by use of frequency-domain measurements of time-dependent photon migration

Frequency-domain studies of time-dependent light propagation in tissuelike phantoms that contain optical heterogeneities are described. Specifically the phase shift and amplitude modulation of reemergent light were measured when illuminated by an amplitude-modulated light source. Changes in the phase angle and the extent of modulation revealed the presence of a light-absorbing object. Furthermore the magnitude and direction of these changes were sensitive to the absorber depth and the light modulation frequency in a manner that could be used to infer the location of the heterogeneity. These data suggest the feasibility of optical imaging by frequency-domain methods.     

Focus groups reveal perils and promises of managed care for nurse practitioners

Decades of practice and research suggest that nurse practitioners (NPs) provide cost-effective and high-quality care. Managed care's emphasis on prevention and cost savings led some policy makers to view NPs as a way to meet the need for primary care providers. However, access to and utilization of NPs has increasingly been controlled by managed care organizations (MCOs) through their selection of providers for primary care panels. This study employed qualitative methodology to examine NPs' experiences with MCOs. Three focus groups, comprising 27 NPs in New York and Connecticut, revealed NPs' mixed reactions to managed care and a range of sentiments regarding NPs' efforts to be listed as primary care providers. The results reflected NPs' concerns about their perceived "invisibility," as well as their sense of "invincibility" in the ways in which NPs are responding to the barriers posed by MCOs. They identified barriers to, as well as ways to facilitate, being listed by MCOs, and described the importance of NPs working individually and collectively in negotiating with MCOs.     

Episodic gonadotropin-releasing hormone gene expression revealed by dynamic monitoring of luciferase reporter activity in single, living neurons

The existence of an intrinsic oscillator for pulsatile gonadotropin-releasing hormone (GnRH) secretion in normal and transformed GnRH neurons raises the question of whether the corresponding gene also is expressed in an episodic manner. To resolve this question, we used a modification of conventional luciferase technology, which enabled continuous monitoring of GnRH gene activity in single, living neurons. With this method, the relative rate of endogenous gene expression is estimated by quantification of photons emitted by individual neurons microinjected with a GnRH promoter-driven luciferase reporter construct. Immortalized GT1-1 neurons, which secrete the decapeptide GnRH in a pulsatile manner conceptually identical to that of their nontransformed counterparts in vivo, were chosen as the model for these studies. First, we injected individual cells with purified luciferase protein and established that the reporter half-life was sufficiently short (50 min) to enable detection of transient changes in gene expression. Next, we subjected transfected GT1-1 cells to continuous monitoring of reporter activity for 16 h and found that the majority of them exhibited spontaneous fluctuations of photonic activity over time. Finally, we established that photonic activity accurately reflected endogenous GnRH gene expression by treating transfected GT1-1 cells with phorbol 12-myristate 13 acetate (a consensus inhibitor of GnRH gene expression) and observing a dramatic suppression of photonic emissions from continuously monitored cells. Taken together, these results demonstrate the validity of our "real-time" strategy for dynamically monitoring GnRH gene activity in living neurons. Moreover, our findings indicate that GnRH gene expression as well as neuropeptide release can occur in an intermittent manner.     

Effect of limbic structure and striatum damage caused by kainic acid on seizure reactions in animals after intracranial injury

It has been established that hippocampus, enthorhinal cortex, amygdala and substantia nigra (pars reticulata) lesions before head injury lead to a decrease of kainic acid-induced behavioral and electrographic seizure expressions. It can be concluded that after head injury the activation of limbic structures excitability due to excitation of "inputs" to these formations takes place. The obtained data indicate the significant role of nucleus caudatus in activation of posttraumatic brain excitatory mechanisms.     

The diagnosis of drug allergies. Utilizing in vitro mast cell test and IgE inhibition test

With the ever-increasing use of pharmaceuticals and the relatively high risk of developing drug allergies, particularly for patients in hospitals and for ambulatory patients with a history of drug allergy, the need to develop in vitro assays for drug allergy is great. In the early 1970's a mast cell technique was developed for diagnosis of drug allergies. A PRIST inhibition assay has also recently been developed to detect IgE antibodies to drug allergens. This test has also been referred to as the Total IgE Inhibition Test by Specific Drug Allergen, and is a variant of the in vitro RAST Test. In vitro mast cell and IgE inhibition tests are applied for identification of drug and chemical allergens and for their cautious clinical trial to prevent future drug and chemical reactions. Over the last eight years, over 1,300 patients were examined utilizing the mast cell technique. Over 100 drugs were tested, with penicillin, barbiturates, "caine" derivatives and sulfonamides most frequently employed. Of 270 patients with well-defined drug reactions, 190 (70 per cent) gave a positive response to the mast cell test. Eighty-five per cent of sera tested with Type I reactions gave a mast cell response. Of these, a group of 30 patients was studied with PRIST inhibition as well. Procedures for comparative testing of necessary drugs and/or chemicals in cases of high anaphylaxis risk of reaction in the clinical setting, hospital or office are included in the study as well as individual case reports. Mast cell assay coupled with IgE inhibition has been successfully used to diagnose drug and chemical allergic reactions. The incidence of positivity is high when the offending drug causes a Type I allergic reaction. The cases reported indicate that both the Mast Cell and the PRIST inhibition assays are useful for diagnosing and setting the clinical treatment and clinical course of the patient. The mast cell assay would be potentially employed for patient use in hospitals where the incidence of drug allergy is highest and for occupational health in the chemical industry. The greatest potential would be in outpatient care applied to patients with multiple drug allergies in the selection of safe drugs (test negative by both methods, and other clinical studies) for future drug usage.     

Coding mammograms using the classification "probably benign finding--short interval follow-up suggested"

OBJECTIVE: Many benign breast lesions revealed by mammography show features indicating that the lesions have a high, but not complete, likelihood of being benign. The Breast Imaging Reporting and Data System (BI-RADS) allows radiologists to classify these mammograms as "probably benign finding-short interval follow-up suggested" (category 3). We explored whether certain factors are associated with the use of category 3 in a national cancer detection program. MATERIALS AND METHODS: We analyzed data from the National Breast and Cervical Cancer Early Detection Program, a comprehensive nationwide program that provides cancer screening for low-income and medically underserved women. The study population included all women at least 40 years old who had undergone mammography on or before September 30, 1996 (n = 372,760). RESULTS: Of the 372,760 mammograms, 7.7% were classified as category 3. The probability of receiving a category 3 classification decreased as patients' ages increased. Women who were symptomatic were nearly twice as likely as women who were asymptomatic to receive a category 3 classification, and women whose clinical breast examinations had abnormal findings were more than twice as likely as women with examinations having normal findings to receive a category 3 classification. The percentage of mammograms classified as category 3 by state or tribal organization ranged from 1.4% to 14.0%. CONCLUSION: Several patient variables, including patient symptomatology, were associated with the probability of having a mammogram classified as category 3. One of the most important determinants was where the patient underwent mammography, which suggests that variability exists among radiologists themselves in using this BI-RADS code for "probably benign" mammographic lesions.