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Antigenic analyses of Sugi basic protein by monoclonal antibodies: I. Distribution and characterization of B-cell-tropic epitopes of Cry j I molecules.

Using 23 monoclonal antibodies raised against Sugi basic protein (SBP, major allergen of Japanese cedar pollen), composed of Cry j I and Cry j II, analyses of B-cell-tropic epitopes of Cry j I were performed. The following results were obtained. (1) As far as the mAbs were used, no major cross-reactive determinants were detected between Cry j I and Cry j II molecules. (2) 21 of the 23 mAbs were specific for Cry j I, and the anti-Cry j II mAbs were classified into four groups by their fine specificities, suggesting that Cry j I bears at least four antigenic determinant regions. (3) Cry j I molecules were found to take a monomeric form in solution and to display no repeating antigenic epitopes on their surfaces. (4) Some of the determinants seemed to be located in the interior of a Cry j I molecule, and when the Ag is coated on a plastic plate, the determinants become exposed on its surface. (5) Binding of human IgE antibodies to Cry j I and Cry j II was blocked by some of the obtained mAbs, suggesting that these epitopes recognized by the mAbs might have an important role in human allergic response against the cedar pollen.

Allergens

Acoustic features and auditory perceptions of the cries of newborns with prenatal and perinatal complications.

This article describes 2 experiments which examine the relation between neonatal cry features and obstetric histories. Experiment 1 shows that 24 clinically healthy, normal newborns who may be at risk due to a high number of prenatal and perinatal complications can be distinguished from 24 low-complications infants by harmonic and durational features of the cry. High-complications infants required more stimulation to elicit the cry, had a longer latency to cry onset, a shorter first cry expiration, a higher cry pitch, and cried less in total time than low-complications infants. In experiment 2, naive adults rated the high-complications infant cries as more aversive, grating sick, urgent, distressing, piercing, discomforting, and arousing than low-complications infant crues. Factor analyses showed that although the low-complications infant cries were described along one dimension of discomfort, a second factor emerged conveying the "sick" nature of the sound of the cry of the high-complications infants. It was suggested that certain cry features may reflect the risk status of the infant.

Adult

The role of auditory stimuli in crying inhibition in the neonate.

An experiment on the latency and duration of the cessation of spontaneous crying supported evidence that a low frequency intense auditory stimulus inhibits crying in neonates. There was a slight increase in the length of the first pause in crying during exposure to a 200-c/s tone, but no difference in the latency of this first pause. Since crying intensity reaches 80 db SPL at the infant ear, it is hypothesized that auditory stimuli juxtaposed during crying is in fact masked. Hence to be effective, an auditory stimulus must be presented at the beginning of a natural pause in crying. An experiment on rhythm in cry patterns did not support the hypothesis that crying may be a naturally rhythmic pattern of behavior analogous to sucking and susceptible to psychophysical investigation. There was a large variance within and between Ss for the mean latency and duration of cry bursts and pauses. Exper. III asked mothers with infants of 10 days, 7 weeks, and 13 weeks of age to keep a crying activity schedule in their own homes. This study revealed a discontinuity between neonates and older infants in the conditions and patterns of crying. Interview data did indicate that reliable positive orienting responses did occur to natural auditory stimuli in a naturalistic setting.

Acoustic Stimulation

Psychophysiological and perceptual responses to infant cries varying in pitch: comparison of adults with low and high scores on the Child Abuse Potential Inventory.

Based on their scores on the Child Abuse Potential (CAP) Inventory, 30 nonparent adults were classified as either having high CAP scores (n = 15) or low CAP scores (n = 15). Subjects' heart rate and skin conductance were assessed as they listened to audio-taped presentations of four "normal" phonated infant cries and four high-pitched, hyperphonated infant cries. Subjects also rated the cries on several perceptual items. Results showed that both groups of subjects found hyperphonated cries more aversive, distressing, urgent, arousing, and sick than phonated cry sounds. Similarly, all listeners' showed higher skin conductance levels (SCL) in response to hyperphonated than phonated cries. SCL became attenuated over time in response to phonated, but not hyperphonated, cries. Adults in the high-CAP group, showed marginally higher heart rates than low-CAP adults following presentation of all infant cry sounds and responded to phonated cries with marginally higher skin conductance levels than low-CAP adults. The responses of high-CAP nonparent adults are similar to those of parents with histories of physically abusive interactions with their infants.

Adult

Discrimination of infants' cry-signals.

A technique for clustering infants' cry-signals on the basis of perceptually discriminable acoustic characteristics is reported. A cry-sequence containing all pairwise combinations of 24 signals was constructed. These signals have been used previously in cry-recognition studies (e.g., Wasz-Hockert, et al., 1968) and consist of six each of pain, hunger, pleasure, and birth. 20 musically competent subjects were tested individually and instructed to compare each cry with the one immediately preceding it in the sequence. Forced-choice "similarity" scores were summed for every pair of cries. These "similarity scores" were data for both a manual and a computerized clustering method. There was absolute agreement between these two methods, and the clusters of cry-signals correspond very closely to the four original cry groups. This seems a valid technique for examining the acoustic similarity of infants' cry-signals.

Adolescent

Pain cry in full-term asphyxiated newborn infants correlated with late findings.

115 pain-induced cries from 45 full-term newborn infants with pre- and perinatal asphyxia were analyzed by sound spectrographic methods. All the infants had signs of intrauterine asphyxia and Apgar score of 6 or less at 5 min. The mean birth weight was 3170 g. The pain cries were recorded before the age of 8 days, 83% of the cries before 3 days of age. The cry analysis was compared with the pain cries of 75 full-term, healthy newborn infants of corresponding birth weight and gestational age. The results showed significant differences between the cries of the asphyxiated newborn and the healthy infants. The duration of the phonation was shorter, the maximum and minimum pitch of the fundamental frequency was significantly higher. Bi-phonation and vibrato occurred more often, double harmonic break and glottal roll less often. An increase in rising, falling-rising and flat types of melody was observed. Retrospectively, the cries were more abnormal if the infant was found to be neurologically damaged at t he check-up at 2-8 years.

Asphyxia Neonatorum

Pattern of night waking and crying of Korean infants from 3 months to 2 years old and its relation with various factors.

Night waking and crying are very common in infancy and can cause problems for parents and families. This study surveyed 218 healthy Korean infants from 3 months to 2 years old to determine their night waking and crying patterns. On average, 83% awoke and 28% cried more than once per night. Ninety-eight percent of the babies slept with a member of the family. Infants with transitional objects or nighttime habits (e.g., finger sucking, touching and/or playing with mother's or own hair, touching a part of mother's or own body) cried more frequently. In terms of the maternal response toward the crying babies, most of the mothers used traditional methods, such as patting/holding, feeding, changing diapers. Only one mother ignored the crying baby, and none used medication, delayed response, or white noise. Sixteen percent of the mothers indicated that their babies' crying constituted a problem for them.

Circadian Rhythm

Spectrum analysis of the cry sounds of well-nourished and malnourished infants.

The cry sounds of 12 well-nourished and 12 malnourished male infants were compared using behavioral and acoustic measures. The cry of the malnourished infant had an initial longer sound, higher pitch, lower amplitude, more arrhythmia, and a longer latency to the next cry sound than the cry of the well-nourished infant. The similarity betweenthe cry of the malnourished infant and the cry of the brain-damaged infant suggested tha malnutrition may affect the regulatory function of the central nervous system. This hypothesis was supported by additional findings which showed that the abnormal cry patterns in the malnourished infants were associated with a low-level orienting response to a pure tone stimulus as measured by the magnitude of heart-rate deceleration.

Acoustic Stimulation

Cry analysis in infants with severe malnutrition.

Sound spectrographic investigations of the cries of 5 infants, age 7 m to 2 y, with severe malnutrition (one with kwashiorkor and four with marasmus) were compared with the cries of 15 healthy children of corresponding age. The cry of the child with kwashiorkor resembled those of the normal infants. The cries of the marasmic children showed a significant increase in the minimum and maximum pitch, and the occurrence of biphonation and flat melody types. These features have also been found in the cries of children with brain damage. We therefore believe that cry and analysis can be an additional means of investigating to what degree the brain is affected in children with malnutrition.

Child, Preschool

Antigenic analyses of Sugi basic protein by monoclonal antibodies: II. Detection of immunoreactive fragments in enzyme-cleaved Cry j I.

The 4 anti-Cry j I mAbs showing an epitope specificity different from each other, 046, 029, 026 and 027, were selected to analyze the structure of the antigenic determinant for each mAb on a Cry j I molecule. Immunoreactive fragments in enzyme-cleaved Cry j I were detected by means of the adsorption on the mAb column and of the binding to the mAbs on Elisa. The mAb 026 was found to be reactive to the fragments containing a Cry j I N-terminal region obtained by V8 protease or pepsin digestion, but not to those by lysylendopeptidase digestion. The mAb 027 was found to be capable of binding to the fragments containing a linear structure of Asn-Ala-Gly-Val-Leu-Thr-Cys-Ser-Leu-Ser-Lys, which were generated by V8 protease, lysylendopeptidase or pepsin digestion. Furthermore, the synthetic peptide Asn-Ala-Gly-Val-Leu-Thr-Cys-Ser- Leu-Ser-Lys-Arg could bind to 027, but not to 026, and could inhibit the binding of 027 to Cry j I or to its immunoreactive fragments. No fragments capable of reacting to the mAbs 046 and 029 could be found in this study, suggesting that 046 and 029 recognize a conformationally constituted epitope of Cry j I molecule which is destroyed by enzymatic cleavage. The epitope recognized by the mAbs 027 or 026 was found to be located in conformationally hidden parts of the molecule which was exposed to react to the mAbs only after the physicochemical or enzymatic treatment.

Allergens

Bacillus thuringiensis pathogenicity islands encode regulatory circuits controlling insecticidal Cry toxin expression during vegetative growth.

Bacillus thuringiensis (Bt) produces insecticidal toxins, including Cry and Vip3 proteins, that are widely used for biological pest control. Cry proteins are classically expressed during sporulation under the control of sporulation-specific σ factors, whereas Vip3 is produced during vegetative growth, suggesting distinct regulatory pathways. Notably, many cry and vip3A genes are clustered within pathogenicity islands (PAIs), such as BtPAI-1. However, whether these PAIs also encode regulatory mechanisms coordinating toxin expression remains unclear. Here, we identify VipR, a BtPAI-1-encoded transcriptional regulator, as an activator of insecticidal gene expression during the vegetative phase in Bt strains HD-1 and CT-43. In these strains, VipR promotes the transcription of BtPAI-1 associated insecticidal genes, including vip3A and selected cry genes, resulting in premature Cry protein accumulation and increased insecticidal activity. In addition, VipR contributes to the vegetative-phase expression of the non-BtPAI-1 cry9Aa genes in strain BGSC 4AE1. Phylogenetic analysis revealed that vipR is widely distributed in one-third of Bt strains, and is strongly associated with PAIs. Futhermore, heterologous expression of vipR in BGSC 4J5 and HD-73 was sufficient to activate vegetative-phase transcription of some cry independently of sporulation-specific σ factor cascade. These results support a role for VipR in coordinating vegetative-phase expression of insecticidal genes in the Bt strains examined and suggest that BtPAI-1 can encode both insecticidal determinants and regulatory functions that influence their expression. These findings provide new insights into the regulatory architecture of Bt pathogenicity islands and may facilitate the engineering of strains with enhanced insecticidal activity.

Bacillus thuringiensis

Spectrographic analysis of pain cry in neonates with cleft palate.

52 phonations of 13 cleft palate neonates were analyzed by sound spectrographic methods. 17 phonetical attributes were included in the study and the first signal after the pain stimulus was analyzed. The cries of the cleft palate infants were compared with the crying of 75 normal babies of the same age. No change in the fundamental frequency, melody type and duration of the cries was seen in association with these anatomical defects. Two of the characteristics studied, vibrato and the 'tonal pit', occurred significantly more often in cries of the cleft palate infants than in cries of the control series. The changes in the qualities seen in association with cleft palate and/or cleft lip do not mimic the abnormalities produced by brain damage.

Cleft Lip

[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

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