Acute myocarditis as a revealing clue of complete Kawasaki disease.

Not available.

Production of human pro-relaxin H2 in the yeast Pichia pastoris.

BACKGROUND: Initially known as the reproductive hormone, relaxin was shown to possess other therapeutically useful properties that include extracellular matrix remodeling, anti-inflammatory, anti-ischemic and angiogenic effects. All these findings make relaxin a potential drug for diverse medical applications. Its precursor, pro-relaxin, is an 18 kDa protein, that shows activity in in vitro assays. Since extraction of relaxin from animal tissues raises several issues, prokaryotes and eukaryotes were both used as expression systems for recombinant relaxin production. Most productive results were obtained when using Escherichia coli as a host for human relaxin expression. However, in such host, relaxin precipitated in the form of inclusion bodies and, therefore, required several expensive recovery steps as cell lysis, refolding and reduction. RESULTS: To overcome the issues related to prokaryotic expression here we report the production and purification of secreted human pro-relaxin H2 by using the methylotrophic yeast Pichia pastoris as expression host. The methanol inducible promoter AOX1 was used to drive expression of the native and histidine tagged forms of pro-relaxin H2 in dual phase fed-batch experiments on the 22 L scale. Both protein forms presented the correct structure, as determined by mass spectrometry and western blotting analyses, and demonstrated to be biologically active in immune enzymatic assays. The presence of the tag allowed to simplify pro-relaxin purification obtaining higher purity. CONCLUSIONS: This work presents a strategy for microbial production of recombinant human pro-relaxin H2 in Pichia pastoris that allowed the obtainment of biologically active pro-hormone, with a final concentration in the fermentation broth ranging between 10 and 14 mg/L of product, as determined by densitometric analyses.

Acute Exercise Increases Sex Differences in Amateur Athletes' Risk Taking.

The research presented here investigates the interaction between acute exercise, biological sex and risk-taking behavior. The study involved 20 amateur athletes (19-33 years old), 10 males and 10 females, who were asked to undergo subsequent experimental sessions designed to compare their risky behaviors on the Balloon Analogue Risk Task (BART) 34 at rest and while exercising at moderate intensity (60% of their maximal aerobic power). Results showed that physical exercise affected male and female participants differently: Whereas males became more risk seeking, females became more risk averse during exercise.

Phenomenological theory of structural relaxation based on a thermorheologically complex relaxation time distribution.

The aim of this work is to explore the consequences on the kinetics of structural relaxation of considering a glass-forming system to consist of a series of small but macroscopic relaxing regions that evolve independently from each other towards equilibrium in the glassy state. The result of this assumption is a thermorheologically complex model. In this approach each relaxing zone has been assumed to follow the Scherer-Hodge model for structural relaxation (with the small modification of taking a linear dependence of configurational heat capacity with temperature). The model thus developed contains four fitting parameters. A least-squares search routine has been used to find the set of model parameters that fit simultaneously four DSC thermograms in PVAc after different thermal histories. The computer-simulated curves are compared with those obtained with Scherer-Hodge model and the model proposed by Gómez and Monleón. The evolution of the relaxation times during cooling or heating scans and also during isothermal annealing below the glass transition has been analysed. It has been shown that the relaxation times distribution narrows in the glassy state with respect to equilibrium. Isothermal annealing causes this distribution to broaden during the process to finally attain in equilibrium the shape defined at temperatures above Tg.

Compositional mapping of chicken chromosomes and identification of the gene-richest regions.

'Compositional chromosomal mapping', namely the assessment of the GC level of chromosomal bands, led to the identification, in the human chromosomes, of the GC-richest H3+ bands and of the GC-poorest L1+ bands, which were so called on the basis of the isochore family predominantly present in the bands. The isochore organization of the avian genome is very similar to those of most mammals, the only difference being the presence of an additional, GC-richest, H4 isochore family. In contrast, the avian karyotypes are very different from those of mammals, being characterized, in most species, by few macrochromosomes and by a large number of microchromosomes. The 'compositional mapping' of chicken mitotic and meiotic chromosomes by in-situ hybridization of isochore families showed that the chicken GC-richest isochores are localized not only on a large number of microchromosomes but also on almost all telomeric bands of macrochromosomes. On the other hand, the GC-poorest isochores are generally localized on the internal regions of macrochromosomes and are almost absent in microchromosomes. Thus, the distinct localization of the GC-richest and the GC-poorest bands observed on human chromosomes appears to be a general feature of chromosomes from warm-blooded vertebrates.

Telomeres in warm-blooded vertebrates are composed of GC-rich isochores.

We have hybridized the vertebrate telomeric sequence (TTAGGG)n on DNA compositional fractions from 13 mammalian species and 3 avian species, representing 9 and 3 orders, respectively. Our results indicate that the 50- to 100-kb fragments derived from telomeric regions are composed of GC-rich and GC-richest isochores. Previous works from our laboratory demonstrated that single-copy sequences from the human H3 isochore family (the GC-richest and gene-richest isochore in the human genome) share homology with compositionally correlated compartments of warm-blooded vertebrates. This correlation suggested that the GC-richest isochores are, as in the human genome, the gene-richest regions of warm-blooded vertebrates' genome. Moreover, this evidence suggests that telomeric regions are the most gene-dense region of all warm-blooded vertebrates. The implications of these findings are discussed.

Phenolphthalein induces chromosome aberrations in human and Chinese hamster liver cells (CHEL) cultured in vitro.

Phenolphthalein is a nonprescription laxative agent that has been widely used during this century. Recent studies in animal models have shown that phenolphthalein has carcinogenic activity. In order to assess cytogenetic effects on human cells in vitro, we tested phenolphthalein in a chromosome aberration assay in human embryo cells derived from amniotic fluid. Our results show that phenolphthalein induces a significant increase in the frequency of chromosome aberrations in human cells. The lowest dose level at which the clastogenic effect is evident is 23.2 microg/ml. Similar positive results were obtained in a Chinese hamster liver cell line, which is metabolically competent to activate different classes of promutagens and procarcinogens into biologically active metabolites. Instead, parallel experiments in Chinese hamster ovary cells did not show any clastogenic effect due to phenolphthalein. These latter data suggested that phenolphthalein acts as a promutagen and must be metabolically activated to exert its clastogenic effect. Teratogenesis Carcinog. Mutagen. 20:209-217, 2000.

Gene density in the Giemsa bands of human chromosomes.

The human genome is formed by isochores belonging to five families, L1, L2, H1, H2 and H3, that are characterized by increasing GC levels and gene concentrations. In-situ hybridization of DNA from different isochore families provides, therefore, information not only on the correlation between isochores and chromosomal bands, but also on the distribution of genes in chromosomes. Three subsets of R(everse) bands were identified: H3+, H3* and H3-, that contain large, moderate, and no detectable amounts, respectively, of the gene-richest H2 and H3 isochores, and replicate very early and early, respectively, in S phase of the cell cycle. Here, we investigated the GC levels, replication timings and DNA compaction of G(iemsa) bands. We showed that G bands comprise two different subsets of bands, one of which is predominantly composed of L1 isochores, replicates at the end of the S phase, has a higher DNA compaction relative to H3+ bands and corresponds to the darkest G bands of Francke (1994). In contrast, the other subset is composed of L2 and H1 isochores, has less-extreme properties in replication and composition and corresponds to the less-dark G bands of Francke.

[Not Available]. / Vito Volterra organizzatore scientifico e la nascita della biologia matematica in Italia.

In the last few decades quite a few articles have been published concerning Volterra's contributions to biomathematics. These articles are usually focused on the works of Volterra published after 1926, while they overlook Volterra's intense activity as scientific organizer in the field of oceanographic studies in the years that span from 1910 - the year of his celebrated dissertation about the applications of mathematics to social and biological sciences - to 1925, when he met his prospective son-in-law Umberto D'Ancona. In this article I shall try to fill this gap by telling the history of Volterra's scientific collaboration with Italian biologists (such as Giovan Battista Grassi, Luigi de Marchi and Gustavo Brunelli) in the decades before his meeting with Umberto D'Ancona. This story is worth telling because it helps to explain the stronger emphasis Volterra placed in his first biomathematical papers on the importance of the applications of mathematics to biology for practical matters (such as the prevention of epidemics and the enhancement of the fishing industry) rather than on more theoretical issues, which he tackled only in the last years of his life.

Compositional mapping of mouse chromosomes and identification of the gene-rich regions.

The mouse genome is a mosaic of isochores, consisting of long (> 300 kb), compositionally homogeneous DNA segments that can be divided into two GC-poor families, L1 and L2, representing 56% of the genome, and two GC-rich families, H1 and H2, representing 26% and 7% of the genome, respectively, the remaining 11% being formed by satellite and ribosomal DNAs. (GC is the molar fraction of guanine + cytosine in DNA.) The mouse genome differs from the human genome (which is representative of most mammalian genomes) because it shows a narrower compositional spectrum of isochores and it has a karyotype formed exclusively by acrocentric chromosomes. The chromosomal distribution of the four isochore families, as investigated here by in situ hybridization of single-copy sequences from compositional DNA fractions, has shown that G(lemsa) bands are essentially composed of GC-poor isochores, whereas R(everse) bands comprise three subsets of bands: R' bands, containing GC-poor isochores and GC-rich isochores of the H1 family, and T and T' bands, containing all H2 isochores (in addition to other isochores), the former containing a higher proportion of H2 isochores than the latter. Mouse T and T' bands are generally syntenic with, and are compositionally related to, human T and T' bands and have the highest gene concentrations. These findings indicate that the distribution of isochore families and genes in chromosomal bands is basically similar in mouse and in human genomes, in spite of their remarkable differences and their extremely large phylogenetic distance.

Identification of the gene-richest bands in human chromosomes.

The human genome is a mosaic of isochores, long DNA segments which are compositionally homogeneous and which can be partitioned into five families, L1, L2, H1, H2 and H3, characterized by increasing GC levels and by increasing gene concentrations. Previous investigations showed that in situ hybridization with a DNA fraction derived from the GC-richest and gene-richest isochores of the H3 family produced the highest concentration of signals on 25 R(everse) bands that include the 22 most thermal-denaturation-resistant T(elomeric) bands, a subset of R bands. Using an improved protocol for in situ hybridization and cloned H3 isochore DNA, we have now shown (i) that the number of bands which are characterized by strong hybridization signals, and which are here called T or H3+, is 28; (ii) that 31 additional R bands, here called T'or H3* bands, also contain H3 isochores, although at a lower concentration than H3+ bands; and (iii) that the remaining R bands (about 140 out of 200, at a resolution of 400 bands), here called R" or H3- bands, do not contain any detectable H3 isochores. H3+ and H3* bands contain all the gene-richest isochores of the human genome. The existence of three distinct sets of R bands is further supported (i) by the different compositional features of genes located in them; (ii) by the very low gene density of chromosomes 13 and 18, in which all R bands are H3- bands; (iii) by the compositional map of a H3* band, Xq28; (iv) by the overwhelming presence of GC-rich and GC-poor long (> 50 kb) DNA sequences in H3+/H3* and in H3-/G bands, respectively; and (v) by the large degree of coincidence of H3+ and H3* bands with CpG island-positive bands. These observations have implications for our understanding of the causes of chromosome banding and provide a classification of chromosomal bands that is related to GC level (and to gene concentration).

Predicting maltreatment of children of teenage mothers.

OBJECTIVE: To determine the degree to which knowing certain characteristics about young high-risk families can help distinguish those families most likely to maltreat their children from those families at lower risk of maltreating their children. DESIGN: Observational cohort from which the following predictor variables were gathered when infants were 2 months old: maternal age, depressive symptoms, childrearing attitudes, social support, and living situation (with or apart from related adults). Families were followed up for 24 months to identify the occurrence of maltreatment. SETTING: An urban, socioeconomically disadvantaged cohort of teenage mothers and their infants attending a hospital-based special primary care clinic for teen mothers and their infants. PARTICIPANTS: All full-term infants and mothers enrolled into the clinic in 1990 participated in the study. This included 47 mother-infant pairs enrolled when infants were 2 months of age. Forty-five of these pairs were available for follow-up when infants were 24 months of age. MAIN OUTCOME MEASURES: Maltreatment defined as any incident that prompted investigation by the state child protective agency and was found to be a substantiated case of maltreatment by that agency. RESULTS: Maltreatment occurred in 15 of 45 families before the child's second birthday. Discriminate function analysis produced a model that correctly classified 13 of 15 maltreating mothers and misclassified one of 30 non-maltreating mothers. Stepwise analysis revealed that living situation was by far the strongest predictive variable (R2 = 7). CONCLUSION: Maltreatment was a predictable outcome within this extremely high-risk cohort. Living apart from related adults was the strongest risk factor associated with maltreatment. This easily obtainable piece of information may be an important risk marker for practitioners, social service personnel, and others working with this very-high-risk population. It may allow early supportive interventions that might prevent maltreatment.

Evaluation of cytogenetic effects of a naturally occurring non-ice-nucleation Pseudomonas fluorescens strain in Chinese hamster ovary (CHO) cells.

One of the main methods for eliminating ice-nucleation-active (INA+) bacteria the micro-organisms responsible for frost injuries to plants at mild freezing temperatures, is the use, as competitors, of other naturally occurring non-nucleating strains (non-INA). In the present article we investigated the cytogenetic effects of a naturally occurring non-INA strain of Pseudomonas fluorescens (MS 1640 R3), evaluating the induction of chromosomal aberrations and sister chromatid exchanges (SCEs) in Chinese hamster ovary (CHO) cells in the absence and presence of rat S9 metabolism. The results obtained did not show any increase in either chromosomal aberrations or SCEs, both in the absence and presence of rat S9 metabolism when used as i) intact bacteria cells, ii) sonicated bacteria (i.e., potential endotoxins), or iii) metabolic bacterial products (i.e., potential exotoxins) released in the growth medium.

Possible association between fluoxetine hydrochloride and colic in an infant.

This is a case study of a 6-week-old infant referred for colic whose mother was using fluoxetine hydrochloride and breast feeding the infant. The mother switched to a commercial formula and reported a dramatic decrease in the infant's crying. We asked the mother to feed the infant breast milk from a bottle and she agreed. Throughout the study the mother kept a daily diary of her infant's crying, sleeping, stooling patterns, and feeding problems. Analysis of the mother's breast milk showed concentrations of 69 ng/mL for fluoxetine and 90 ng/mL for norfluoxetine. Infant blood serum/plasma level was analyzed for fluoxetine hydrochloride following return to breast milk. The concentrations were 340 ng/mL for fluoxetine and 208 ng/mL for norfluoxetine. The diary records showed increased crying, decreased sleep, increased vomiting, and watery stools when fluoxetine hydrochloride was transmitted through breast feeding or breast milk in bottle. These symptoms were reduced when the infant was formula fed. We suggest a possible relationship between colic and associated symptoms and fluoxetine hydrochloride in maternal breast milk.