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PubMed1997

Cloning and analysis of the first cry gene from Bacillus popilliae.

An 80-kDa parasporal crystal protein was detected in protein extracts of sporangia of Bacillus popilliae isolated from a diseased larva of the common cockchafer (Melolontha melolontha L.). Amino acid analysis of tryptic peptides revealed significant homology to the Cry2Aa endotoxins of Bacillus thuringiensis. The gene cryBP1 (cry18Aa1), which codes for the parasporal crystal protein, was found in a putative cry operon on the bacterial chromosome, which contains at least one further (smaller) open reading frame, orf1. The 706-amino-acid-long CryBP1 (Cry18Aa1) protein has a predicted molecular mass of 79 kDa and shows about 40% sequence identity to the Cry2 polypeptides of B. thuringiensis. In the light of published observations which suggest that the parasporal crystal proteins of B. popilliae are slightly toxic to their grub hosts, we propose the following survival strategy of B. popilliae. As an obligate pathogen of grubs, B. popilliae germinates in the gut of a grub and the parasporal crystal proteins are released and activated. The activated protein does not cause colloid osmotic lysis but instead damages the gut wall somehow to allow the vegetative cells to enter the hemolymph more easily. By becoming a parasite, B. popilliae can continue to proliferate efficiently while the living grub provides a food supply. This process is in contrast to that of B. thuringiensis, which rapidly kills the insect and is then limited to growth on the larval carcass.

Amino Acid Sequence↗
PubMed2001

Nuclear entry mechanism of rat PER2 (rPER2): role of rPER2 in nuclear localization of CRY protein.

Mammalian PERIOD2 protein (PER2) is the product of a clock gene that controls circadian rhythms, because PER2-deficient mice have an arrhythmic phenotype. The nuclear entry regulation of clock gene products is a key step in proper circadian rhythm formation in both Drosophila and mammals, because the periodic transcription of clock genes is controlled by an intracellular, oscillating, negative feedback loop. The present study used deletion mutants of rat PER2 (rPER2) to identify the functional nuclear localization signal (NLS) in rPER2. The elimination of putative NLS (residues 778 to 794) from the rPER2 fragment resulted in the loss of nuclear entry activity. Adding the NLS to the cytosolic protein (bacterial alkaline phosphatase) translocates the fusion protein to the nuclei. The data indicate the presence of a functional NLS in rPER2. Furthermore, intact rPER2 was preferentially translocated from the cytoplasm to the nucleus when coexpressed with human CRY1 (hCRY1). However, rPER2 mutants lacking a carboxyl-terminal domain could not enter the nucleus even in the presence of hCRY1. In addition, coexpression of the nuclear localization domain (residues 512 to 794) lacking rPER2 and CRY1 changed the subcellular localization of CRY1 from the nucleus to the cytoplasm. In vitro protein interaction studies demonstrated that the carboxyl-terminal domain of rPER2 is essential for binding to CRY1. The data suggested that both the rPER2 NLS and carboxyl-terminal CRY binding domain are essential for nuclear entry of the rPER2-CRY1 complex.

Active Transport, Cell Nucleus↗
PubMed2000

Don't cry for us Argentinians: two decades of teaching medical humanities.

Medical humanities--history, literature, anthropology, ethics and fine arts applied to medicine--play an important role in medical education. For more than 20 years an effort has been made to obtain an academic identity for such a multidisciplinary approach. A distinction between humanitarianism and humanism is attempted here, the former being associated with medical care and the latter with medical education. In order more precisely to define the relationship between the arts and medicine, an alternative term "medical kalology", as-yet-unsanctioned, coined after the rules of medical terminology, is proposed. The Department of Medical Humanities in the School of Medicine, National University La Plata, submits the following apologia: Don't cry for us Argentinians, since the teaching of medical humanities has helped our doctors to function more truly humanistically during the past two decades, as we intend to continue with this calling in the future.

Anthropology↗
PubMed2004

Diversity of cry genes and genetic characterization of Bacillus thuringiensis isolated from Brazil.

Two hundred and eighteen Bacillus thuringiensis isolates from Brazil were characterized by the presence of crystal protein genes by PCR with primers specific to different cry and cyt genes. Among these isolates, 95 were selected according to their geographic origin for genetic characterization with the 16S rRNA gene, RAPD, and plasmid profile. Isolates containing cry1 genes were the most abundant (48%) followed by the cry11 and cyt (7%) and cry8 genes (2%). Finally, 40.3% of the isolates did not produce any PCR product. The plasmid profile and RAPD analysis showed a remarkable diversity among the isolates of B. thuringiensis not observed in the 16S rRNA gene. These results suggest that the genetic diversity of B. thuringiensis species results from the influence of different ecological factors and spatial separation between strains generated by the conquest of different habitats.

Bacillus thuringiensis↗
PubMed2003

Reduction of antigenicity of Cry j I, major allergen of Japanese cedar pollen, by the attachment of polysaccharides.

An attempt was made to mask the allergenic structure of a major allergen protein, Cry j I (CJI), in Japanese cedar pollen using the Maillard-type polysaccharide conjugation. The SDS-PAGE pattern of the CJI-galactomannan conjugate prepared by the Maillard reaction showed broad bands widely distributed from 50 kDa to more than 100 kDa, suggesting the attachment of galactomannan. The competitive enzyme-linked immunosorbent assay showed that the IgE antibody in the sera of cedar pollen-sensitive patients reacted strongly with CJI, while it did not react with the CJI-galactomannan conjugate. This result suggests that the antigenicity of CJI is greatly reduced by the conjugation with galactomannan.

Allergens↗
PubMed1999

[A study of aggravation of atopic dermatitis during Japanese cedar pollen season--correlation with grades of dermatitis on face and Cry j 1 specific IgE].

We studied influence of Japanese cedar pollen (Jcp) on aggravation of atopic dermatitis (AD) during the pollination season. 48.5% of 97 patients with atopic dermatitis showed aggravation of dermatitis during the pollination season and 85% of them had Japanese cedar pollinosis, whereas only 44% of AD patients without the aggravation had the pollinosis. There was no difference of grades of dermatitis on face between the groups with or without the aggravation. Furthermore, we measured specific IgE to Jcp and Cry j 1, a major allergen of Jcp, by ELISA in the sera from the 54 patients with AD. The levels of specific IgE antibodies to both allergens in the group with the pollinosis were significantly higher than in the group without the pollinosis. However, significant difference of those was not recognized between the groups with or without the aggravation of AD. Therefore, our study has suggested that Japanese cedar pollen is likely to be one of causes of seasonal aggravation of AD in individuals sensitized to the pollen, and some other factors, e.g. Jcp-specific T cells, might play an important roll in addition to the Jcp-specific IgE.

Adolescent↗
PubMed2002

[Identification of airborne pollen and airborne particles with pollen allergen (Cry j 1, Dac g) by aeroallergen immunoblotting technique].

After collection of airborne particles with a seven-day recording volumetric spore trap (Burkard model) using optically clear, pressure sensitive acrylic adhesive tapes, a dry PVDF membrane was pressed down firmly onto the adhesive tape, antigen-antibody reaction was performed in full contact with the tapes and PVDF membrane. Dark purple spots from airborne pollen allergens were examined under a light microscope to evaluate the form of pollens and particles wit h the antigenicity. It is clarified that identification of the form of pollens with the antigencity is possible not only pollens with the antigenicity but a pollen has already lost its shape, and it is also clarified that some airborne particular matters have the pollen antigencity. Airborne samples were collected during the Cryptomeria japonica or grass pollen season. Samples collected during the C. japonica pollen season were treated with anti-Cry j 1 monoclonal antibody, and those during the grass pollen season with anti-Dac g rabbit IgG. Samples collected during the C. japonica and the grass pollen peak season were treated with antibodies from sera of pollinosis patients. Spots originated not only from relevant pollen grains but also from parts of airborne particular matter have the pollen antigenicity and some spots could not observed for any kind of particles.

Air Pollutants↗
PubMed2002

[Characterization of cry gene and broad spectrum against lepidopteran of Bacillus thuringiensis subsp. colmeri 15A3].

Bacillus thuringiensis wild type strain 15A3 belongs to subspecies colmeri serotype H-21. RFLP and PCR analysis show that it contains six types of ICP genes: cry1Aa, cry1Ac, cry1Ca, cry1D, cry1I and cry. The sequence of the 1.45 kb N-terminal fragment of cry1Aa differed from that of published. SDS-PAGE showed that the crystal consists of proteins with molecular weight about 130, 79, 70, 65, 51 and 45 kD. Strain 15A3 didn't sysnthesize heat-stable beta-exotoxins according to test of house fly aberration. The 1.2 tons fermentative production exhibited high toxicity against three lepidopteran pests: H. armigera, S. exigua and H. cunea. It was proved that wild type strain can produce a broad specturm of ICP.

Animals↗
PubMed2004

Crisis of faith vs. spiritual cry of distress.

Traumatic events radiate shock waves that impact the totality of an individual: the physical, cognitive, emotional, behavioral and spiritual aspects of a person. Symptoms of distress appear horizontally in relationships with family, friends, and co-workers and vertically in relationship with God. People who have a faith relationship with God will often use religious language to express their anguish and despair regarding a traumatic event. The challenge of any crisis interventionist is to be able to hear accurately the cry of the distressed individual and respond so as to connect with and instill hope in the traumatized individual.

Crisis Intervention↗
PubMed2005

[Crying upon eating: the crocodile-tears syndrome].

A male infant who, since birth, had begun to cry as soon as he began to nurse was found to have the crocodile tears syndrome. It is thought that in this condition the lacrimal glands are partially innervated by efferent fibres of the facial nerve (VII). The syndrome may be congenital, but may also be a consequence of an infection or trauma. Treatment is surgical or by the use of botulinum-A toxin.

Botulinum Toxins, Type A↗
PubMed2005

Generation of mouse-human hybridomas secreting human monoclonal antibodies to Japanese cedar pollen allergen Cry j1.

Peripheral blood lymphocytes from 13 donors were transformed with Epstein-Barr virus (EBV) to establish immortalized human B-cells secreting antibodies to Japanese cedar (Cryptomeria japonica) pollen allergens. EBV-transformed B-lymphocytes were then fused with mouse myeloma SP2/O3, and three clones of mouse-human hybridomas secreting human IgM class monoclonal antibodies to a cedar pollen allergen Cry j1 were established.

Allergens↗
PubMed1994

The effects of trypsin on ATP-sensitive potassium channel properties and sulfonylurea receptors in the CRI-G1 insulin-secreting cell line.

The effects of the proteolytic enzyme trypsin upon ATP-sensitive potassium (KATP) channel activity were examined in the CRI-G1 insulin-secreting cell line. Trypsin activated channels only when applied to the intracellular surface of the cell membrane. The activation could be prevented by the concomitant application of trypsin inhibitor or by heat inactivation of the enzyme. The trypsin-induced change in channel activity was accompanied by a reduction in the rate of channel rundown. However, trypsin did not affect the mean single channel conductance (55.2 pS), the ionic selectivity, or rectification of the KATP channel. Concentration response curves for various KATP channel inhibitors were constructed in the presence and absence of intracellular trypsin. The EC50 for tolbutamide was shifted from 30.0 +/- 4.5 microM, with 100 micrograms/ml heat-inactivated trypsin present to 9.7 +/- 1.0 mM with active trypsin in the intracellular solution. Treatment of the cells' external surface with 1 mg/ml trypsin did not alter the potency of tolbutamide. Intracellular trypsin also produced a significant fall in the potency of glibenclamide, meglitinide, and phentolamine but did not alter the effectiveness of thiopentone. Radioligand binding studies demonstrated a total loss of 3H-labeled glibenclamide binding when the intracellular surface of the cells was exposed to trypsin. In contrast, 3H-labeled glibenclamide binding was not affected when the enzyme was applied to the external surface. Trypsin treatment, therefore, alters a number of characteristics of KATP channel pharmacology, and we suggest that this is due to action at possibly more than one site but includes the functional cleavage of the sulfonylurea receptor from the KATP channel.

ATP-Binding Cassette Transporters↗
PubMedDate not supplied

Asymmetric crying facies: an index of other malformations.

Congenital hypoplasia or absence of the depressor anguli oris muscle is a minor congenital anomaly causing asymmetrical crying facies (ACF). The interesting aspect of this abnormality lies in the frequently associated abnormalities. Cardiac, urogenital, musculoskeletal, respiratory and cervicofacial defects have been described in cases with ACF. Therefore it is suggested that ACF can be used as an index of other congenital malformations. We report three cases with ACF who had varied congenital anormalites.

Abnormalities, Multiple↗
PubMed1996

Antigenic analysis of Cryptomeria japonica and Chamaecyparis obtusa using anti-Cry j 1 monoclonal antibodies.

In Japan, pollen of Cryptomeria japonica and Chamaecyparis obtusa are a yearly source of distress for many people suffering seasonally from allergic rhinitis. To study common epitopes shared by the two species, two monoclonal antibodies (moAbs) were raised against Cry j 1, which is the most predominant allergen in C. japonica. One of the moAbs was found to be reactive even to the major allergen of C. obtusa, demonstrating that the moAb (J1B01) can detect an epitope shared by both species J1B01 strongly inhibited the binding of the major allergens of C. japonica and C. obtusa to IgE of patients who are sensitive to C. japonica and C. obtusa. This finding signifies the importance of the epitope recognized by J1B01.

Allergens↗
PubMed1997

Two patients with asymmetric crying facies, normal cardiovascular systems and deletion of chromosome 22q11.

We report two unrelated patients with Asymmetric Crying Facies (ACF) and 22q11 deletion. In the first case ACF was the presenting feature. Fluorescent In Situ Hybridization (FISH) revealed 22q11 deletion and subsequent echocardiography demonstrated a normal cardiovascular system. The second presented with hypoparathyroidism, shown on FISH to be secondary to 22q11 deletion. Echocardiography was normal and ACF was found incidentally. Deletion of 22q11 has been reported with ACF occurring as part of the Cardiofacial syndrome (CFS) [Giannotti A, Mingarelli R, (1994): Am J Med Genet 53: 303-304] but this is the first report of ACF with 22q11 deletion in the absence of cardiac abnormalities. A review of the literature pertaining to ACF is presented.

Cardiovascular System↗
PubMed1998

Cry j 1-induced synthesis of interleukin-5 and interferon-gamma by peripheral blood mononuclear cells of patients with seasonal allergic rhinitis due to Japanese cedar pollens.

This study comprised 130 adult patients with Japanese cedar pollen-specific IgE in the serum and 15 non-atopic individuals. Eighteen patients had no seasonal aggravation of nasal symptoms during the pollen season in 1998 (asymptomatic group). Forty-two patients had not been treated previously with immunotherapy and were treated with antihistamine tablets during the pollen season in 1998 (medication group). Sixty-one patients had undergone variable periods of immunotherapy using pollen extracts, and they were further divided into a good-IT group who responded markedly to immunotherapy and a poor-IT group who responded poorly to immunotherapy. The remaining nine patients had been treated with immunotherapy for more than 12 years and all of them had stopped immunotherapy by the end of May 1997 because they had no nasal symptoms for the last three pollen seasons and were considered to be cured of seasonal allergic rhinitis (cure group). Peripheral blood mononuclear cells (PBMCs) were collected from each subject during the cedar pollen season in 1998 and were stimulated for 96 h with 10 micrograms/ml Cry j 1. The concentrations of interleukin-5 (IL-5) and interferon-gamma (IFN-gamma) in the culture supernatant were determined using an enzyme-linked immunosorbent assay. The levels of IFN-gamma did not differ significantly among the non-atopic group, the asymptomatic group, the medication group, the poor-IT group and the good-IT group. The level of IL-5 in the asymptomatic group was not different from that in the non-atopic group. The levels of IL-5 in the medication group, the good-IT group and the poor-IT group were significantly higher than in the non-atopic group. The level of IL-5 in the good-IT group, but not in the poor-IT group, was significantly lower than in the medication group. The level of IL-5 in the cure group was not significantly different from in the non-atopic group, and the level of IFN-gamma in the cure group was significantly lower than in the non-atopic group. In conclusion, immunotherapy can decrease the pollen allergen-induced synthesis of IL-5, but not of IFN-gamma, and this immunological modulation is involved in the working mechanism of immunotherapy related to its clinical efficacy. A tolerance or anergy of both TH1 and TH2 cells under allergen stimulation may be an immunological indication of cure after the treatment of seasonal allergic rhinitis. Thus, the suppression of synthesis of IL-5 and IFN-gamma by allergen-stimulated PBMCs is likely to be a reliable criterion for a possible cure of seasonal allergic rhinitis after immunotherapy.

Adult↗