A direct PCR detection method for Clostridium tyrobutyricum spores in up to 100 milliliters of raw milk.

A direct detection method for Clostridium tyrobutyricum spores in up to 100 ml of raw milk is presented. The bacterial spores are concentrated by centrifugation after chemical extraction of the milk components. The vegetative cells are selectively lysed, and their DNA is digested and washed away. Afterwards, the DNA is liberated from the spores by microwave treatment. For the identification of the C. tyrobutyricum DNA, a two-step PCR method with two nested pairs of primers is used. The primers were derived from the 16S-23S rRNA spacer region of C. tyrobutyricum, and the specificity of each of them for C. tyrobutyricum is demonstrated. The detection limit can be estimated to be between 3 and 30 spores in 100 ml of raw milk.

ROMK inwardly rectifying ATP-sensitive K+ channel. II. Cloning and distribution of alternative forms.

The rat ROMK gene encodes inwardly rectifying, ATP-regulated K+ channels [K. Ho, C. G. Nichols, W. J. Lederer, J. Lytton, P. M. Vassilev, M. V. Kanazirska, and S. C. Hebert. Nature Lond. 362: 31-38, 1993; H. Zhou, S. S. Tate, and L. G. Palmer. Am. J. Physiol. 266 (Cell Physiol. 35): C809-C824, 1994], and mRNA encoding these channels is widely expressed in distal cortical and outer medullary nephron segments [see companion study; W.-S. Lee and S. C. Hebert. Am. J. Physiol. 268 (Renal Fluid Electrolyte Physiol. 37): F1124-F1131, 1995]. Using approaches based on homology to ROMK1, we have identified two additional ROMK isoforms, ROMK2b and ROMK3. Analysis of the nucleotide sequences of the ROMK isoforms indicates that molecular diversity of ROMK transcripts is due to alternative splicing at both the 5'-coding and 3'-noncoding regions. The splicing at the 5' end of ROMK gives rise to channel proteins with variable-length NH2 termini containing different initial amino acid sequences. Functional expression of these isoforms in Xenopus oocytes showed that they form functional Ba(2+)-sensitive K+ channels. The nephron distribution of mRNAs encoding alternatively spliced isoforms of ROMK (ROMK1-ROMK3) was investigated by reverse transcription-polymerase chain reaction (RT-PCR) of nephron segments dissected from rat kidney. Nondegenerate PCR primer pairs were designed to span at least one intron and to amplify specific alternatively spliced forms of ROMK.(ABSTRACT TRUNCATED AT 250 WORDS)

Direct detection of Listeria monocytogenes in 25 milliliters of raw milk by a two-step PCR with nested primers.

A method for direct detection of Listeria monocytogenes in 25 ml of raw milk is presented. The detection limit can be situated between 10 and 5 CFU. The detection method is based on chemical extraction of the milk components and PCR amplification with two nested pairs of primers specific for Listeria monocytogenes.

Personality antecedents of depressive tendencies in 18-year-olds: a prospective study.

Antecedents of depressive tendencies at age 18 were longitudinally evaluated using data from nursery school through high school. Depression was measured by CES-D scores from which the contribution of self-reported anxiety was partialed. As early as age 7, boys who subsequently acknowledged dysthymia were aggressive, self-aggrandizing, and undercontrolled whereas girls with later depressive tendencies were intropunitive, oversocialized, and overcontrolling. Similar gender differences were observed in pre- and early adolescence. At age 14, dysthymic boys were more likely to use both marijuana and harder drugs whereas dysthymic girls showed no tendency to use marijuana but did show a marked tendency to experiment with hard drugs. These girls also displayed low self-esteem. Preschool IQ correlated positively with dysthymia in girls and negatively in boys. The psychodynamics of gender differences in depressive affect were discussed.

Parental agreement on child rearing during early childhood and the psychological characteristics of adolescents.

In previous research, an index of parental agreement regarding child-rearing orientations was found to relate to the quality of children's psychological functioning from 3 to 7 years of age. For this study, the parental agreement index, derived when their children were 3 years old, was related to a variety of psychological measures and personality descriptions obtained when the children were adolescents. Reliable differences were found in the patterning of relations for boys and girls wherein parental agreement during early childhood was associated with tested intelligence, aspects of moral judgment, and dimensions of personality derived from self-reports for boys but not for girls. For adolescent girls, early parental agreement was associated with the congruence of "self" and "ideal-self" descriptions (i.e., self-esteem). For both girls and boys, parental agreement was associated with personality descriptions provided by observers; however, it was only in the sample of girls that early parental agreement significantly correlated with Q sort criterion scores referencing ego-undercontrol and ego-resiliency. These findings suggest that early family socialization experiences are importantly but differently salient for boys and girls with respect both to psychological content and the developmental timing of effects. The data suggest that socialization researchers consider the likelihood that developmental paths to competence differ markedly for girls and boys.

Longitudinally foretelling drug usage in adolescence: early childhood personality and environmental precursors.

Drug usage in early adolescence (age 14) was related to concurrent and preschool personality characteristics for a sample of 54 girls and 51 boys. The personality concomitants and antecedents of drug use differed somewhat as a function of gender and the drug used. At age 14, for both sexes, the use of marijuana was related to ego undercontrol, while the use of harder drugs reflected an absence of ego-resiliency, with undercontrol also a contributing factor. At ages 3/4, subsequent adolescent drug usage in girls related to both undercontrol and lower ego-resiliency. In boys, adolescent drug usage related strongly, during their nursery school years, to undercontrol and with resiliency having no long-term implications. Early family environment related to adolescent drug usage in girls but not in boys. Drug usage in adolescent girls was related to homes earlier identified as unstructured and laissez-faire, where there was little pressure to achieve. Drug usage related to other substance use and, in boys, to IQ decline from age 11 to age 18. Implications of these results for contemporary views regarding adolescent drug usage are discussed.

Testing aspects of Carl Rogers's theory of creative environments: child-rearing antecedents of creative potential in young adolescents.

Longitudinal data involving 106 children and their parents were used to test preschool child-rearing implications of Carl Rogers's theory of creativity-fostering environments (Rogers, 1954). Indices were developed for each parent and for each mother-father combination that reflected the degree to which the parents' child-rearing practices and interactions with their preschool children matched the recommendations implicit in Rogers's description of a creativity-fostering environment. The three indices of Rogers-prescribed child-rearing practices each correlated positively (rs = .38 to .46) and significantly (all ps less than .001) with a composite index of creative potential in early adolescence, 7 to 11 years later. Rogers-prescribed preschool child-rearing practices also emerged as significant antecedents of adolescent creative potential in regression/path analyses that held constant the influence of sex, preschool intelligence, and preschool creative potential. Theoretical and methodological aspects of the study are discussed.

The personality of children prior to divorce: a prospective study.

In a longitudinal study, the personalities of children from intact families at ages 3, 4, and 7 were reliably assessed by independent sets of raters using Q-items reflecting important psychological characteristics of children. A number of these families subsequently experienced divorce. The behavior of boys was found, as early as 11 years prior to parental separation or formal dissolution of marriage, to be consistently affected by what can be presumed to be predivorce familial stress. The behavior of boys from subsequently divorcing families was characterized by undercontrol of impulse, aggression, and excessive energy prior to parental divorce. The behavior of girls from subsequently divorcing families was found to be notably less affected by the stresses in families prior to parental divorce. The prospective relations afforded by the longitudinal analyses suggest that the behavior of conflicting, inaccessible parents during the preseparation period may have serious consequences for personality development, especially for boys. Hence, some characteristics of children commonly seen to be a consequence of divorce may be present prior to marital dissolution.

Egocentrism and ego resiliency: personality characteristics associated with perspective-taking from early childhood to adolescence.

In this study we examined the longitudinal relation between ego resiliency and egocentrism. Measures of Level 1 and Level 2 egocentrism were administered to one hundred eleven 3 1/2-year-old children. Independent personality evaluations were available on these children at ages 3, 4, 7, 11, and 14 in the form of Q-sort ratings by independent sets of teachers and examiners. The relation between egocentrism and ego resiliency was observed to depend on level of egocentrism, as well as on sex of subject. In the sample of girls, both Level 1 and Level 2 egocentrism were negatively correlated with ego resiliency concurrently at age 3, but no long-term implications of egocentrism emerged. In the sample of boys, Level 1 egocentrism was consistently negatively associated with ego resiliency from early childhood (age 3) into adolescence (age 14). However, Level 2 egocentrism displayed no concurrent nor any dependable longitudinal relation with ego resiliency. The discussion focuses on possible interpretations of the relation between egocentrism and ego resiliency and on the sex differences in the pattern of longitudinal results.

Preparation and characterization of potential prodrugs of dyphylline.

Four diesters and four monoesters of dyphylline were synthesized as prodrugs proposed to prolong the duration of action of dyphylline. They were characterized by IR, 1H NMR, HPLC, and MS. Appropriate solvent-programming conditions for the HPLC separation of dyphylline and the newly synthesized mono- and diesters were developed. It was confirmed by low-temperature 1H NMR at approximately -40 degrees C that all four monoesters were located on the primary hydroxy position. Attempts to produce the secondary monoesters yielded the primary monoesters during purification. Monoesters were shown by HPLC and MS to migrate between the primary and secondary hydroxy groups in aqueous solution.

Differential premises arising from differential socialization of the sexes: some conjectures.

The sex-differentiated socialization emphases of parents and other representatives of societal institutions are considered as they influence the personality development and behavioral orientations of males and females. Specifically, sex-differentiated socialization emphases, "shaping" behaviors, and teaching styles are evaluated with regard to the nature of the "meta-messages" conveyed to boys and girls during their early, formative years. These messages are assumed to differentially influence the self-concepts evolved, ego structures, personal goals, and the cognitive-adaptational heuristics of boys and of girls. Differences in the socialization environments experienced by the 2 sexes can be seen as related to gender differences in personality characteristics. To integrate the empirical findings surrounding gender differences in personality and socialization experience, some conjectures are offered regarding the different self- and world views our current culture may be creating and fostering in males and in females. The potential and even likely influence of biological factors conjoined with the bidirectional effects of child and parent interaction are recognized as confounded with an interpretation in terms of differential socialization. But also, it is noted that until the effects of differential socialization are specifically evaluated by cultural, subcultural, or individual family changes, the role of biological and bidirectional factors cannot be assessed.

Pattern recognition of steroids using fragment molecular connectivity.

The use of pattern recognition methods to classify a set of steroids into five therapeutic categories was investigated. First-order fragment molecular connectivity values were determined for 10 positions on each molecule using a template-based method of position assignment. Learning set and test set classifications were performed. Although the numbers of compounds misclassified were comparable for all of the methods, the identities of the misclassified compounds varied depending on whether the classification method assumed a local or a global view of the data. The best classification results were comparable to those obtained by linear and quadratic discriminant analyses. For this set of compounds, it was concluded that pattern recognition methods offer no advantages over traditional discriminant analysis methods if classification alone is considered, especially since most discriminant analysis procedures utilize stepwise variable selection, which is not as common in pattern recognition analyses.

Classification of drugs by discriminant analysis using fragment molecular connectivity values.

An investigation was made into the use of linear and quadratic discriminant analysis, along with K nearest-neighbor analysis, in the classification of a set of 51 compounds which were divided into five therapeutic categories. By superimposing each compound on a pattern structure, as first proposed by Cammarata, eight positions were assigned on the molecule. Each position was coded with the numerical value of a descriptor index. Relative molar refraction, which was the index used by Cammarata, was compared with a number of molecular connective indices. For each of the indices studied, it was found that only four of the eight positions contributed significantly to between-class differences. It was also found that first-order molecular connectivity, calculated as the sum of the contributions of each of the bonds joining a given position, resulted in consistently fewer misclassifications as compared with the other indices. Using first-order molecular connectivity, validation procedures were performed on the original set of compounds, on random samples drawn from this set, and on a set of ten compounds not included in the analysis. The results obtained were highly data dependent, but they, nevertheless, suggest that molecular connectivity indices should prove useful in structural classification procedures.

Paired-ion reversed-phase high-pressure liquid chromatographic assay of pentobarbital-pyrilamine suppositories.

The assay of suppositories containing pentobarbital and/or pyrilamine in a water-soluble polyethylene glycol base by high-pressure liquid chromatography is described. No extraction is required. The suppository is dissolved in the mobile phase. This solution is diluted with an internal standard stock solution containing phenobarbital. Chromatographic conditions include a C18 bonded microporous silica column and a mobile phase of 65% 4 x 10(-3) M n-butyl sodium sulfonate in 1% acetic acid and 35% acetonitrile. The procedure using commercial products gave results comparable to those obtained by GLC.