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Contents of cry genes and insecticidal toxicity of Bacillus thuringiensis strains from terrestrial and aquatic habitats.

AIMS: Two Bacillus thuringiensis collections from terrestrial and aquatic habitats were compared in order to study the possible interrelationships between habitat and biological characteristics (serovar, cry genes content and toxicity). METHODS AND RESULTS: Bacillus thuringiensis strains were characterized by serology, PCR, and one-dose treatment against the noctuids Helicoverpa armigera and Spodoptera exigua, and the dipteran Tipula oleracea. A total of 12 and 10 different serovars were identified within terrestrial and aquatic strains, respectively. The number of non-toxic strains was greater in aquatic (41.6%) than in terrestrial habitats (5.3%). The genes cry1C, cry1D and cry1E were significantly more frequent in the terrestrial habitat. The cry1B gene was very frequent within thuringiensis strains. CONCLUSIONS: A high diversity was found in terms of serovars present and cry genes content in both collections. The relative frequency of individual cry genes was different in both collections, and a serovar-dependent distribution of the cry1B gene was found. Some strains sharing the same set of cry genes differed in their toxicity, suggesting important differences in gene expression. SIGNIFICANCE AND IMPACT OF THE STUDY: The inter-relationships between serology, cry gene content and toxicity may allow a better understanding of B. thuringiensis ecology.

Animals↗

Immunodominance of seven regions of a major allergen, Cry j 2, of Japanese cedar pollen for T-cell immunity.

The immunodominant regions of the Japanese cedar pollen allergen Cry j 2 for T-cell immunity were determined with whole peripheral blood lymphocytes (PBL) derived from seven allergic patients and three nonallergic subjects. Cry j 2-stimulated T-cell proliferation was inhibited by anti-HLA-DR, but not by anti-HLA-DQ antibody, indicating that the responding T cells recognized the allergen peptides associated with HLA-DR molecules. It was found that seven regions of Cry j 2, i.e., regions corresponding to amino acid numbers 1-26, 70-84, 151-167, 187-203, 252-279, 283-314, and 345-362, were immunodominant for T-cell proliferation. Thus, Cry j 2 bears a limited number of immunodominant regions despite polymorphic features of HLA-DR in the immune system. This suggests the possibility of molecularly designing Cry j 2 antagonists that could downregulate allergic reactions to Japanese cedar pollen.

Case-Control Studies↗

New strategy for identification of novel Cry-type genes from Bacillus thuringiensis strains.

We designed five degenerate primers for detection of novel cry genes from Bacillus thuringiensis strains. An efficient strategy was developed based on a two-step PCR approach with these primers in five pair combinations. In the first step, only one of the primer pairs is used in the PCR, which allows amplification of DNA fragments encoding protein regions that include consensus domains of representative proteins belonging to different Cry groups. A second PCR is performed by using the first-step amplification products as DNA templates and the set of five primer combinations. Cloning and sequencing of the last-step amplicons allow both the identification of known cry genes encoding Cry proteins covering a wide phylogenetic distance and the detection and characterization of cry-related sequences from novel B. thuringiensis isolates.

Bacillus thuringiensis↗

Parallel computation of genetic likelihoods using CRI-MAP, PVM, and a network of distributed workstations.

We have developed a version of the CRI-MAP computer program for genetic likelihood computations that runs the FLIPS and ALL functions of CRI-MAP in parallel on a distributed network of workstations. The performance of CRI-MAP-PVM was assessed in several linkage analyses using the FLIPS option of CRI-MAP on a map of 85 microsatellite markers for human chromosome 1. These analyses showed excellent speedup and efficiency and low distribution overhead. In addition, we have adapted the MultiMap program for automated construction of linkage maps to use CRI-MAP-PVM. These improvements significantly reduce the time required to compare likelihoods of different marker orders. Thus, the construction of linkage maps can proceed in a more timely fashion, in keeping with recent advances in genotyping technology.

Algorithms↗

Antigenicity of the oligosaccharide moiety of the Japanese cedar (Cryptomeria japonica) pollen allergen, Cry jI.

Cry jI, a major allergenic glycoprotein of Cryptomeria japonica (Japanese cedar, Sugi), is the most common pollen allergen in Japan. Carbohydrate analysis and lectin staining indicated that Cry jI possesses the fucose/xylose-containing N-linked oligosaccharide which previously has been found in some plant glycoproteins. Rabbit polyclonal anti-Cry jI IgG antibodies were found to be highly cross-reactive with two other plant glycoproteins which have the same type of oligosaccharides, and the cross-reactivities were completely abolished on chemical deglycosylation of the glycoproteins. Enzyme-linked immunosorbent assay inhibition showed that the majority (up to 70%) of the anti-Cry jI rabbit IgG antibodies recognized the oligosaccharide moiety of Cry jI. The carbohydrate epitopes were found to be only partially involved in the binding of specific IgE antibodies from a pool of human patient sera.

Allergens↗

The crayfish Procambarus clarkii CRY shows daily and circadian variation.

The circadian rhythms of crayfish are entrained by blue light, through putative extra retinal photoreceptors. We investigated the presence and daily variation of CRY, a protein photosensitive to blue light spectra and ubiquitous in animals and plants, in the putative pacemakers of Procambarus clarkii, namely the eyestalk and brain, at different times of the 24 h light:dark cycles. Using different experimental light protocols and by means of qualitative/quantitative immunofluorescence anatomical and biochemical methods, we identified CRY immunoreactivity in cells located in the medulla-terminalis-hemiellipsoidal complex (MT-HB) and the anterior margin of the median protocerebrum (PR). The immunoreaction varied with the time of day and the two neural structures showed a semi-mirror image. The results of the biochemical analysis matched these variations. Western blotting demonstrated statistically significant circadian rhythms in brain CRY abundance, but no daily circadian CRY abundance oscillations in the eyestalk. These immunocytochemical and biochemical results link a specific photoreceptor molecule to circadian rhythmicity. We propose that CRY may be linked to the photoreception of the clock and to the generation of circadian rhythmicity.

Animals↗

[An enzyme-linked immunosorbent assay for the quantitation of the major allergen from Japanese cedar (Cryptomeria japonica) pollen, Cry j I, using monoclonal antibodies].

We have developed an enzyme-linked immunosorbent assay (ELISA) using two anti-Cry j I monoclonal antibodies (KW-S10 and KW-S131) for the quantitation of the major allergen from Japanese cedar (Cryptomeria Japonica) pollen, Cry j I. Polystyrene microplates coated with KW-S131 were incubated with pollen allergen extracts. Cry j I, which bound to the antibody, was detected with alkaline phosphatase-conjugated KW-S10 using chromogenic enzyme substrate. Cry j I could be measured in concentration of between 0.16 and 2.5 ng/ml by this assay. Intra- and inter-assay coefficients of variation were 1.1-3.5% and 0.9-4.6%, respectively. This assay was considered that it was specific for Cry j I of Japanese cedar pollen because it didn't react with allergens of hinoki (Chamaecypairs obtusa) pollen which have antigenicity in common to Japanese cedar pollen. This assay would be useful for the standardization of Japanese cedar pollen allergen extracts.

Allergens↗

An ethological analysis of human infant crying: answering Tinbergen's four questions.

The proximate causes, survival value, ontogeny, and evolutionary history of human infant crying are examined. Experiments and field observations involving infant distress vocalizations and begging calls in avian, mammalian, and nonhuman primate species are considered, as are ethnographic records of infant care and responses to crying in nonindustrialized societies. It is argued that human infant crying evolved as a primarily acoustic, graded signal, that it is a fairly reliable, if imperfect, indicator of need for parental care and that its primary function is to promote parental caregiving. Selection pressures that may have shaped the evolution of crying and its potential for corruption through dishonesty also are discussed.

Adolescent↗

Crying following stroke. A report on 30 cases.

Following stroke, many patients experience a heightened tendency to cry. The aim of the present study was to provide a detailed description of this behavior and its accompaniments. Thirty stroke patients who had cried in the previous month were studied using a detailed structured clinical interview. There was a wide range of frequency of crying. The components of crying combine in a more complex and varied way than commonly held stereotyped views suggest. No distinct subtypes were identified. Further research is required to determine the relationship between emotionalism and the frequency and severity of symptoms of depression.

Activities of Daily Living↗

Acoustic features of normal-hearing pre-term infant cry.

Acoustic features of expiratory cry vocalizations were studied in 125 pre-term infants prior to being discharged from a level-3 neonatal intensive care unit. The purpose was to describe various phonatory behaviors in infants in whom significant hearing loss could be ruled out. We also compared these results with normal-hearing full-term infants, and evaluated whether linkage exists among acoustic cry features and various anthropometric, diagnostic and treatment variables obtained throughout the peri- and neonatal periods. Our analysis revealed that cry duration was significantly related to total days receiving respiratory assistance. The occurrence of other complex spectral and temporal aspects of acoustic cry vocalizations including harmonic doubling and vibrato also increased in infants receiving some form of respiratory assistance. The presence of harmonic doubling also depended on weight and conceptional age at test. The discussion focuses on the implication of these relationships and directions for future research.

Acoustics↗

Prenatal and perinatal lead exposures alter acoustic cry parameters of neonate.

We performed acoustic analyses on cries elicited from a subset of healthy babies born to the Mexico City Prospective Lead Study at 2 days (n = 75), 15 days (n = 176), and 30 days (n = 166). Lead was measured in maternal blood every 8 weeks during pregnancy from week 12 to delivery and in umbilical cord (1-38 micrograms/dL, 0.05-1.84 mumol/L). Percent nasalization and number of cries decreased in babies born to mothers with higher lead levels in the last two trimesters while median fundamental frequency increased in babies born to mothers with higher lead at 12 weeks of pregnancy, and with higher cord lead in multiple regression analysis. Decreased percent nasalization was related to increased brainstem auditory evoked response latencies and interpeak intervals in a subset of the sample. The results suggest an effect of gestational exposure to lead on apparatus innervated by cranial nerves and/or lead effect on cry mediated by lead-altered auditory function. Altered baby cry and auditory function associated with lead might contribute to developmental delays by affecting early communication between caretaker and baby.

Acoustics↗

Maternal soothing and infant stress responses: soothing, crying and adrenocortical activity during inoculation.

The relation between maternal soothing and infant stress response during inoculation was examined in a sample of 37 mothers and their 3-month-old infants. The mothers' soothing and the infants' cry vocalizations and the mothers' and the infants' salivary cortisol level pre- and post-injection were analysed. There was a positive relation between infants' cry vocalization post-injection and maternal soothing pre- and post-injection. The sample was divided in two sub-groups depending on whether the mothers evidenced most soothing of the infants in the period before (Preparatory group; n=20) or after (Contingent group; n=17) the syringe injection. In the Preparatory group, the duration of infant cry vocalizations was related to amount of maternal soothing before and after the injection, while cry vocalizations in the Contingent group was related to amount of maternal soothing after the injection. The Contingent infants responded to the injection with a significant increase in cortisol, while there was no increase in the Preparatory infants. The Preparatory infants evidenced significantly longer duration of looking at the target stimuli in a visual marking task, suggesting greater difficulties in disengaging attention. These findings indicate that 3-month-olds' stress responses and their mothers' situational behaviour are mutually regulated.

Adult↗

Psychomotor development in 65 home-reared children with cri-du-chat syndrome.

The psychomotor development of 65 noninstitutionalized individuals with cri-du-chat syndrome was examined through parental questionnaire responses and supporting medical records. Social quotients determined by a Vineland Maturity Scale ranged from 6 to 85, the ages at which developmental milestones were attained varied from the upper limits of normal to six years delayed. Achievement levels were influenced favorably by the early introduction of special education, and were affected adversely by the presence of an unbalanced translocation. This study suggests that many children with cri-du-chat syndrome can attain developmental and social skills normally seen in 5- to 6-year-old children, although their linguistic abilities are seldom as advanced. Contrary to the commonly portrayed clinical picture of severe mental retardation and bedridden debilitation, the older home-reared cri-du-chat child was usually ambulatory, had a moderate degree of independence in self-care skills, and was able to communicate either verbally or through gestural sign language. Physicians and parents should be aware of the full range of psychomotor potential of the child with cri-du-chat syndrome in order to make informed decisions concerning institutional placement.

Achievement↗

Predicting infant crying from fetal movement data: an exploratory study.

Using a standardised procedure, 20 women who were 37 weeks pregnant logged fetal movements for 1 h in the morning and evening for 3 days. Close agreement between such logs and ultrasound recordings of individual differences in fetal movements have been documented. After birth, the babies' sleeping, waking, feeding and crying behaviour were recorded prospectively in validated behaviour diaries at 1, 6 and 12 weeks of age. The fetal movements showed an evening peak, due to a diurnal variation in strong movements. These strong movements did not predict the babies' crying. Weak fetal movements provided a reliable index of fetal differences and predicted the amounts the babies cried at 1, 6 and 12 weeks of age. The findings suggest that temperament-like dispositions present before babies are born predict how much they are likely to cry.

Behavior↗

Audition of laughing and crying leads to right amygdala activation in a low-noise fMRI setting.

Adequate behavioral responses to socially relevant stimuli are often impaired after lesions of the amygdala. These impaired behavioral responses in particular concern the recognition of facial, and sometimes vocal, expressions of fear. Using low-noise functional magnetic resonance imaging (fMRI) in combination with controlled sound delivery, we investigated how the amygdala, insula and auditory cortex are involved in the processing of affective non-verbal vocalizations (laughing, crying) in healthy humans. The same samples of male and female laughing and crying were presented in two different experimental conditions: self-induction of the corresponding emotions while listening, and detection of artificial pitch shifts in the same stimuli. Both conditions led to bilateral activation of the amygdala, insula and auditory cortex with a right-hemisphere advantage in the amygdala, and larger activation during laughing than crying in the auditory cortex with a slight right-hemisphere advantage for laughing, both likely due to acoustic stimulus features. The results show that amygdala activation by emotionally meaningful sounds like laughing and crying is independent of the emotional involvement, suggesting the pattern recognition aspect of these sounds is crucial for this activation. This aspect was revealed by a low-noise fMRI protocol which presumably minimized confounding effects of stressful high-noise fMRI.

Acoustic Stimulation↗

Reduction of crying episodes owing to infantile colic: A randomized controlled study on the efficacy of a new infant formula.

OBJECTIVES: The aim of this study was to evaluate the efficacy on crying episodes owing to infantile colic of a new infant formula containing partially hydrolysed whey proteins, prebiotic oligosaccharides (OS), with a high beta-palmitic acid content. DESIGN: Prospective randomized controlled study. SETTING: Italy. SUBJECTS: Two hundred and sixty-seven formula-fed infants, aged less than 4 months, with infantile colic, were randomized to receive either the new infant formula (study treatment (ST)) or a standard formula and simethicone (6 mg/kg twice a day) (control treatment (CT)). A questionnaire was given to parents to evaluate for 14 days the daily number of colic episodes and crying time. RESULTS: Out of the 199 infants who completed the study, 96 were treated with the new formula and 103 were not treated. Infants receiving the new formula had a significant decrease in colic episodes after 1 week (2.47+/-1.94 at day 7 vs 5.99+/-1.84 at the study entry) compared to infants receiving the CT (3.72+/-1.98 at day 7 vs 5.41+/-1.88 at the study entry) (P < 0.0001). Also at day 14, the crying episodes were significantly different between the two groups of infants (1.76+/-1.60 in ST vs 3.32+/-2.06 in CT) (P < 0.0001). CONCLUSIONS: The use of a partially hydrolysed formula supplemented with fructo- and galacto-OS induces a reduction of crying episodes in infants with colic after 7 and 14 days when compared with a standard formula and simethicone.

Colic↗

The cry characteristics of an infant who died of the sudden infant death syndrome.

Fourteen cries of a four day old infant who subsequently died suddenly of unexplained causes were analyzed on nine acoustic characteristics including fo, duration, formant frequencies and sound pressure level. In comparison to a group of newborn controls, the Sudden Infant Death Syndrome (SIDS) victim's cries exhibited a lower fo, longer duration, lower formant frequencies and greater sound pressure level throughout the spectrum. Cry duration and sound pressure levels, however, deviated in excess on one standard deviation from the mean of the other newborns. Similar findings resulted when the SIDS infant was compared to a group of full term infants who were siblings of SIDS victims, although the magnitude of the differences was slightly less especially with respect to sound pressure level. Measurement of selected acoustic variables in a newborn's cry may be of value in our understanding of SIDS and for identifying infants at risk.

Acoustics↗

Living and coping with excessive infantile crying.

AIM: The aim of the study was to elicit from parents how they lived and coped with a baby who cries excessively. It sought to identify factors which made coping more difficult, as well as interventions perceived by parents to be effective in improving the situation. BACKGROUND: The excessive crying of a baby can be one of the most trying aspects of parenthood. Although there have been many studies into the cause and treatment of the problem, these have produced little convincing evidence and even less agreement among investigators. In the light of this an alternative approach is to provide effective support to enhance coping until the crying abates. DESIGN: This study was based in pragmatic ethnography and followed an adapted grounded theory approach. A total of 25 adults from 14 families participated, recounting their experiences with 20 babies who either were currently crying excessively or had done so previously while less than 1 year old. Data was collected by a factual questionnaire and tape-recorded focused interviews conducted in the family home augmented by periods of participant observation. OUTCOMES: Almost every aspect of family life was disrupted, resulting in strained relationships, feelings of guilt, and concerns about losing control. Repeated attempts had to be made to establish a diagnosis and to find a cure. Such attempts failed, leading to a repeated cycle of hope and disappointment. The greatest potential to effect a positive impact lay in the supportive role of health visitors. They needed to visit frequently; stay for a prolonged period; demonstrate engagement with the family and its difficulties; and impart specific messages with conviction and sincerity. CONCLUSION: A specific, vital role was found for the health visitor in supporting parents through this problem, and aspects of the role were identified in which further study is indicated.

Activities of Daily Living↗