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Nucleotide-dependent activation of KATP channels by diazoxide in CRI-G1 insulin-secreting cells.

1. Patch-clamp recording techniques were used, to examine the effects of diazoxide on KATP currents in CRI-G1 insulin-secreting cells in the presence of non-hydrolysable nucleotides. 2. In the presence of non- or slowly-hydrolyzed ATP analogues, bathing the intracellular aspect of cell-free membrane patches diazoxide inhibited KATP channel activity. 3. Under whole-cell recording conditions, with various non-hydrolysable nucleotides present intracellularly (after dialysis), diazoxide induced KATP current activation. The largest activation occurred with Mg-adenylyl-(beta, gamma-methylene) diphosphate (Mg-AMP-PCP) present in the dialysing solution. This activation was diazoxide- and nucleotide-concentration-dependent. 4. In the absence of Mg2+, or in the presence of manganese (Mn2+) ions intracellularly, diazoxide did not induce KATP current activation, regardless of the species of nucleotide present in the pipette. 5. Intracellularly applied trypsin prevented the activation of KATP currents by diazoxide in the presence of Mg-AMP-PCP, an effect reversed by co-application of intracellular polymethylsulphonyl fluoride with the trypsin. 6. The application, by dialysis, of a CRI-G1 cell lysate, with negligible Mg-ATP, resulted in a substantial activation of the KATP current by diazoxide. 7. It is concluded that diazoxide can activate KATP channel currents by two separate pathways, one requiring a phosphorylation process, the other the presence of an intracellular protein coupled with a Mg-purine nucleotide.

Adenine Nucleotides↗

Interaction of Bacillus thuringiensis svar. israelensis Cry toxins with binding sites from Aedes aegypti (Diptera: Culicidae) larvae midgut.

This work shows in vitro processing of Bacillus thuringiensis svar. isralensis Cry toxins and the capacity of the active fragments to bind the midgut microvilli of Aedes aegypti larvae. Processing of Cry11Aa, Cry4Aa and Cry4Ba yielded double fragments of 38-30, 45-20 and 45-18 kDa, respectively. Competition assays showed that all active (125)I-Cry toxins are able to specifically bind to brush border membrane fractions and they might share a common class of binding sites. The values of IC(50) suggested that toxins do not display high affinity for the receptors from brush border membrane fractions, while dissociation assays showed that binding was irreversible, indicating the insertion of toxins in the cell membrane.

Animals↗

Genetic variability of Spodoptera frugiperda Smith (Lepidoptera: Noctuidae) populations from Latin America is associated with variations in susceptibility to Bacillus thuringiensis cry toxins.

Bacillus thuringiensis strains isolated from Latin American soil samples that showed toxicity against three Spodoptera frugiperda populations from different geographical areas (Mexico, Colombia, and Brazil) were characterized on the basis of their insecticidal activity, crystal morphology, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of parasporal crystals, plasmid profiles, and cry gene content. We found that the different S. frugiperda populations display different susceptibilities to the selected B. thuringiensis strains and also to pure preparations of Cry1B, Cry1C, and Cry1D toxins. Binding assays performed with pure toxin demonstrated that the differences in the toxin binding capacities of these insect populations correlated with the observed differences in susceptibility to the three Cry toxins analyzed. Finally, the genetic variability of the three insect populations was analyzed by random amplification of polymorphic DNA-PCR, which showed significant genetic diversity among the three S. frugiperda populations analyzed. The data presented here show that the genetic variability of S. frugiperda populations should be carefully considered in the development of insect pest control strategies, including the deployment of genetically modified maize in different geographical regions.

Animals↗

Characterization of cry genes in a Mexican Bacillus thuringiensis strain collection.

Mexico is located in a transition zone between the Nearctic and Neotropical biogeographical regions and contains a rich and unique biodiversity. A total of 496 Bacillus thuringiensis strains were isolated from 503 soil samples collected from the five macroregions of the country. The characterization of the strain collection provided useful information on the ecological patterns of distribution of B. thuringiensis and opportunities for the selection of strains to develop novel bioinsecticidal products. The analysis of the strains was based on multiplex PCR with novel general and specific primers that could detect the cry1, cry3, cry5, cry7, cry8, cry9, cry11, cry12, cry13, cry14, cry21, and cyt genes. The proteins belonging to the Cry1 and Cry9 groups are toxic for lepidopteran insects. The Cry3, Cry7, and Cry8 proteins are active against coleopteran insects. The Cry5, Cry12, Cry13, and Cry14 proteins are nematocidal. The Cry11, Cry21, and Cyt proteins are toxic for dipteran insects. Six pairs of general primers are used in this method. Strains for which unique PCR product profiles were obtained with the general primers were further characterized by additional PCRs with specific primers. Strains containing cry1 genes were the most abundant in our collection (49.5%). Thirty-three different cry1-type profiles were identified. B. thuringiensis strains harboring cry3 genes represented 21.5% of the strains, and 7.9% of the strains contained cry11 and cyt genes. cry7, cry8, and cry9 genes were found in 0.6, 2.4, and 2.6% of the strains, respectively. No strains carrying cry5, cry12, cry13, cry14, or cry21 genes were found. Finally, 14% of the strains did not give any PCR product and did not react with any polyclonal antisera. Our results indicate the presence of strains that may harbor potentially novel Cry proteins as well as strains with combinations of less frequently observed cry genes.

Animals↗

[Daily life and work: conceptions of chronic renal insufficiency (CRI) patients and their relatives].

This descriptive and exploratory research aimed to know the conceptions, attitudes and behaviors about work as reported by 16 chronic renal insufficiency patients and their relatives. Data were collected from April to August 2000 through semi-structured interviews. The results revealed that work is valued by all families as a source of health and financial resources, besides being important for individuals' character formation; that most CRI patients do not carry out any remunerated activities and, when they do, they count on the help and understanding of the boss and relatives. We conclude that CRI and its treatment do not directly or absolutely impede the realization of this kind of activity, but entail important limitations. We consider it is necessary for health professionals to join in the search for support by relatives and society, so that these people can be inserted in the labor market when they want to and are in the right conditions.

Activities of Daily Living↗

Asymmetric crying facies: the cardiofacial syndrome.

A 5 1/2 month old female child presented with cyanosis and recurrent respiratory infection. Asymmetric facies was noticed on crying. On investigation an ostium secundum atrial septal defect with right partial anomalous pulmonary venous connection and severe pulmonary hypertension was detected. The association of asymmetric crying facies with congenital cardiac anomalies, known as cardiofacial syndrome, is presented here to highlight the use of this clinical sign as a marker to congenital anomalies.

Cyanosis↗

[A new counting method of airborne Japanese cedar (Cryptomeria japonica) pollen allergens by immunoblotting using anti-Cry j I monoclonal antibody].

We produced monoclonal antibodies against the major allergen of Japanese cedar (Cryptomeria japonica) pollen, Cry j I. KW-S10 antibody reacted only with Japanese cedar pollen and KW-S91 antibody reacted to most angiospermae pollens as well as Japanese cedar pollen. Using these antibodies, we devised a new counting method of Japanese cedar pollen allergen particles by an immunoblotting technique. Airborne pollen allergens were collected on vaseline coated glass slides or Burkard's sampling tape and were transferred onto polyvinylidene difluoride (PVDF) membranes. The membranes were treated with anti-Cry j I monoclonal antibody conjugated with alkaline phosphatase. Pollen allergens were detected as spots on the membranes after staining with phosphatase substrate (BCIP/NBT). This method using KW-S10 antibody measured only the amounts of allergen from Japanese cedar pollen, while with KW-S91 antibody, the method measured the amounts of pollens which have antigenicity in common with Japanese cedar pollen.

Allergens↗

Definitive localization of X-linked Kallman syndrome (hypogonadotropic hypogonadism and anosmia) to Xp22.3: close linkage to the hypervariable repeat sequence CRI-S232.

Kallmann syndrome is a genetically heterogeneous disease characterized by hypogonadotropic hypogonadism and anosmia. Six families in which the disorder followed an X-linked inheritance were investigated by linkage analysis. Diagnostic criteria were uniformly applied and included tests for hypogonadotropic hypogonadism and anosmia. Close linkage was found by using the hypervariable repeated sequence CRI-S232 (DXS278) previously mapped to Xp22.3. At a maximum lod score of 6.5, the recombination fraction was calculated as .03. Of 30 fully informative meioses, one recombination between the disease locus and the loci recognized by probe CRI-S232 was observed. When an independent approach is used, these results confirm the X-linked Kallmann syndrome assignment previously made by deletion mapping, and allow definitive localization of the syndrome assignment previously made by deletion mapping, and allow definitive localization of the syndrome to the Xp22.3 region. This opens the way to carrier detection and to the identification of a gene responsible for this disorder.

Chromosome Mapping↗

[Recognition of onomatopoeias and animal cries during focalized lesions of the cerebral cortex (author's transl)].

The recognition of animal cries and their onomatopoeias was compared during the course of right or left unilateral cortical lesions. A double dissociation is noted; in left-sided affections there is a more severe deficiency to onomatopoeia than to cries, whereas in right-sided lesions the quality of the performances is reversed. These results suggest that the right hemisphere is dominant in the auditive recognition of familiar sounds, and supply information as to the linguistic value of onomatopoeias.

Animals↗

[Spectral analysis of a "sea gull cry" murmur (author's transl)].

"Precordial honk" is a loud, high pitched, musical and usually intermittent systolic noise. The concept of extracardiac origin and "inocence" of this finding is superseded and today its valvular origin during mitral prolapse, mitral or tricuspidal insufficiency - due to rheumatic or ischaemic disease - or during cardiomiopathy is demonstrated. The acoustic characterization of this finding was based on phonocardiographic registration. All the authors describe the "precordial honk" as a high frequency murmur without specifying the acoustic characteristics of the phenomenon. A spectral analysis technique applied to the acoustic cardiac signal (Fourier analysis) has been employed in order to study the "precordial honk" of a 15-year-old boy with mitral valve prolapse. This made possible to show that this sound had only one fundamental frequency at 300 Hz. without harmonics; a "sea gull cry" murmur (as the cry of many birds is) justifying the name given by clinicians. The spectral analysis of phonocardiography signal allow to study, more precisely, the cardiac acoustic phenomenon.

Adolescent↗

[Airborne Japanese cedar allergens studied by immunoblotting technique using anti-Cry j I monoclonal antibody--comparison with actual pollen counts and effect of wind speed and directions].

We collected airborne particles of Japanese cedar pollen with Burkard's sampling tape in Toyama from February to April 1992. The tape was cut into two pieces in parallel to time axis. The one of piece of the tapes was stained with glycerin-jerry and stained pollens were counted with a microscope. The other piece was treated according to the immunoblotting technique. The airborne pollen allergens, reacting with anti-Cry j I monoclonal antibody, were stained as blue spots. The spots were classified by diameter into two groups, large spots (> 50 microns) and small spots (< 50 microns). There were significant correlations found between the airborne Cry j I allergen spots (in large and small) and actual pollen counts obtained with the Burkard's sampler and the Durham's sampler (r = 0.729, 0.586 in large spots and r = 0.676, 0.489 in small spots, p < 0.001). The counts of small spots stayed in high level even in April when actual pollen counts decreased. We concluded that this discrepancy was caused by allergenic crushed cedar pollen particles staying floating longer than actual pollens. Secondly we set a gauge of wind speed and direction at the same point as the samplers. The actual pollen counts and large spots counts were significantly larger in the wind (SE wind in Toyama city) from cedar trees blooming area than other areas. However small spots counts did not differ significantly according to wind directions. Wind speed did not effect on actual pollen counts, large spots counts and small spots count.

Air↗

rhGH use in children with CRI and undergoing dialysis post-transplant in Japan: a multicentre study. MultiCenter Study Group Japan.

We evaluated the effect of rhGH at both 0.5 and 1.0 i.u./kg/week, on the growth of 83 CRI (pre- and dialysis) and 23 transplanted children after one year. Growth velocity (GV) was significantly increased in predialysis children on a dose of 0.5 i.u. (p < 0.001), from 3.8cm/year at baseline to 7.8cm/year at six months and 7.3cm/year at one year. On a dose of 1.0 i.u., GV was increased from 4.5cm/year at baseline to 10.3cm/year at six months and 8.7cm/year at one year (p < 0.01). When the increment in GV was compared a significant difference was noted between 0.5 i.u. and 0.1 i.u. at both six and 12 months (p < 0.01). GV was also significantly increased in dialysed children treated with 0.5 i.u., from 3.5cm/year to 5.6cm/year at six months (p < 0.01) and 5.4cm/year at one year (p < 0.01). On a dose of 1.0 i.u., the increase was from 3.4 to 8.4cm/year at six months (p < 0.001) and 8.3cm/year at one year (p < 0.01). There was a significant difference in GV between the 0.5 and 1.0 i.u. groups at both six and 12 months (p < 0.01). In transplanted children, GV was significantly improved over one year of GH treatment from 5.0 to 7.7cm/year at 0.5 i.u. (p < 0.05) and from 3.7 to 6.3cm/year at 1.0 i.u. (p < 0.05); however, there was no statistical difference between the groups. Seven out of 23 children showed evidence of deterioration of graft function during rhGH treatment. This was due to acute rejection (AR) in every case. AR was noted in two of 10 children (20%) in the 0.5 i.u. group and five of 13 children (38%) in the 1.0 i.u. group. Our results demonstrate that rhGH effectively stimulates GV, not only in CRI, but also in dialysed children. Attention should, however, be paid to deterioration of renal function.

Child↗

Airway pressures during crying in healthy infants.

Maximal inspiratory and expiratory airway pressures (PI max and PE max) were measured in 100 healthy infants (51 males, 49 females; age range, 0.06-3.76 years) by occluding the airway with a suitable face mask during a crying effort. Mean values +/- SD for PI max and PE max were 118 +/- 21 cm H2O and 125 +/- 35 cm H2O, respectively. Maximal inspiratory pressure was independent of age, sex, and anthropometrics, while maximal expiratory pressure showed a low but statistically significant positive correlation with body weight (P less than 0.001).

Age Factors↗

A Y/5 translocation in a 45,X male with cri du chat syndrome.

In a patient described as a 45,X male with cri du chat syndrome, combined cytogenetic and molecular methods revealed Y euchromatic material to be translocated onto the short arm of one chromosome 5, resulting in a chromosome der(5)(5qter----5p14::Yp11.31----Ypter). The translocated Y euchromatin comprised only the distal short arm including the pseudoautosomal region and the so-called deletion intervals 1 and 2. A review of 45,X males from the literature showed that; most of them carry a paternally transmitted Y/autosome translocations; resulting in various autosomal deletions. Depending on the segment concerned, the deletion led to congenital malformations.

Chromosome Banding↗

Partial translocation of NOR and its activity in a balanced carrier and in her cri-du-chat fetus.

Cytogenetic analysis of a balanced reciprocal translocation t(5;13) carrier and her unbalanced 5p- conceptus was carried out. This carrier mother had previously given birth to a child with cri-du-chat syndrome. Silver staining demonstrated the breakpoint on 13p within the nucleolus organizer region (NOR). In the carrier, the NORs both at the original site (13p) and at the translocated site (5p) were silver-stained, indicating that the rRNA genes at both sites were transcribed. The NOR at the derivative chromosome 5 was also silver-stained in the fetus.

Adult↗

Acoustic analysis of pathological cries, stridors and coughing sounds in infancy.

In the examination of respiratory pathological sound phenomena in infancy (pathological crying, stridor and coughing sounds), besides the subjective-auditive observations and traditional examination methods (X-ray, endoscopy), acoustic analysis, as a new branch of bioacoustics, may be of much help. The various pathological infant vocalizations originating from the air passageways are merely symptoms, but they may furnish important diagnostic clues. Approximately 200 recordings of pathological sound signals derived from 180 infants suffering from 40 different diseases or anomalies were analyzed by means of sound spectrography, the fundamental frequency meter, and rarely, the minimal-time-interval spectra technique. The acoustical evaluation of the sound phenomena are not only of scientific and documentative value; they are also of great help in diagnostics and prophylaxis, in the suitable choice of diagnostic tools and in education and further training, i.e. in the everyday work of the pediatric otolaryngologist.

Acoustics↗