Immunostimulants and prevention of recurrent respiratory tract infections.

Recurrent respiratory tract infections (RRTIs) are very common in children and a major challenge for pediatricians. They affect the children's quality of life, cause absences from school and lost parental working days, and repeated medical examinations, hospital admissions as well as antibiotic therapies lead to high costs for society. Given their prevalence and clinical importance, various prevention strategies have been developed. One of the most widely used is the administration of immunostimulants: i.e. molecules of bacterial or synthetic origin that interact with immunological mechanisms in vitro and in vivo. A number of studies have investigated their effects on cellular and innate immunity, and their clinical efficacy, but there is no consensus as to their real usefulness. The main aim of this review is to analyse the available data concerning the activity and efficacy of immunostimulants in preventing pediatric RRTIs. The majority of studies have shown that the number of infections decreases after immunostimulant treatment, but they are affected by various methodological weaknesses. Further studies are urgently needed to confirm whether, when and which immunostimulants should be used.

Vermian subtentorial arteriovenous malformation supplied by the artery of Wollschlaeger and Wollschlaeger.

BACKGROUND: The artery of Wollschlaeger and Wollschlaeger is a tentorial branch of the superior cerebellar artery: due to its small diameter, it is not usually seen in normal angiograms except when enlarged in the setting of a dural AVF or tentorial meningioma. Its presence has been rarely described in the Literature. CASE REPORT: herein we describe the first ever reported case of a vermian subtentorial arteriovenous malformation supplied by the artery of Wollschlaeger and Wollschlaeger in 70 year old female patient. CONCLUSION: vermian subtentorial AVMs supplied by the artery of Wollschlaeger and Wollschlaeger are extremely rare vascular malformations. The presence of the artery of Wollschlaeger and Wollschlaeger must be carefully evaluated during preoperative surgical planning due to its key role in the supply of vascular malformation and to decrease the risk of intra operative bleeding during surgery.

Establishment and characterization of new mammary adenocarcinoma cell lines derived from double transgenic mice expressing GFP and neu oncogene.

A new murine cell line, named GFPneu, was established from a mammary adenocarcinoma arising in double transgenic MMTVneu x CMV-GFP mice. Breast tumours develop in 100% of females after 2 months latency, as a result of the over-expression of the activated rat neu oncogene in the mammary glands. All tissues, and in particular the breast tumours, express the GFP protein. This cell line was tumorigenic when inoculated into nude mice and the derived tumours showed the same histological features as the primaries from which they were isolated. Their histopathology reproduces many characteristics of human breast adenocarcinomas, in particular their ability to metastasize. The GFP marker allows us to visualize the presence of lung metastases in fresh tissues immediately, to confirm the histopathology. From a lung metastatic fluorescent nodule, we derived a further cell line, named MTP-GFP, which we also characterized. These two cell lines could be useful to study the role played by the neu oncogene in the maintenance of the transformed phenotype, in the metastatic process, to test novel therapeutic strategies to inhibit primary tumour growth and to observe the generation of distant metastases.

Enhanced expression of common fragile site with occupational exposure to pesticides.

The expression of common fragile sites (FS), induced by aphidicolin, in subjects with occupational history of exposure to pesticides has been studied. Results showed a higher frequency of FS in exposed subjects; in particular, there was an elevated expression of FS at the cancer breakpoints 3p14, 5q31, 7q22, 7q32, 14q24, and 16q22, involved in leukemias and non-Hodgkin's lymphoma. Moreover, the frequency of breaks in chromosomal bands carrying oncogenes or tumor suppressor genes involved in aberrations was significantly higher in exposed subjects at sites 1q25, 3p25, 7p22, 8q24.1, and 13q14.

No statistical association between fragile sites and constitutional chromosome breakpoints.

Ten thousand four hundred ninety-two constitutional breakpoints available from the cytogenetic literature were analyzed for their coincidence with known fragile sites (FS) at 303-band resolution. In this analysis we have taken into account the stochastic connections of some features of chromosome bands with both the presence of FS and constitutional breakage. Our results suggest that there is no particular association between FS and constitutional chromosome rearrangements.

Specific chromosomal aberrations correlated to transformation in Chinese hamster cells.

Cytogenetic changes were investigated during the spontaneous progression of CHEF18 Chinese hamster cells towards tumorigenicity. We further report the chromosomal characterization of a series of spontaneous anchorage-independent clones, as well as of a series of tumor-derived cell lines resulting from injection of late passage cells in nude mice. The high karyotypic homogeneity (presence of four marker chromosomes strictly associated in all the metaphases analyzed) in all clones and tumor-derived cell lines prompted us to alter the specific pattern of chromosomal aberrations in order to identify which if any of the aberrations were more strictly related to transformation. For this purpose we treated a tumor-derived cell line with Colcemid and analyzed the reversion of anchorage-independent phenotype in the subclones showing an altered association of the four marker chromosomes. We conclude that two of four marker chromosomes contribute to anchorage independence.

Aphidicolin-sensitive specific common fragile sites: a biomarker of exposure to pesticides.

In this work, we analyzed the aphidicolin-sensitive common fragile sites in seven females and four males occupationally exposed to pesticides and in ten controls. The same males had been monitored one year earlier in a previous study by the same authors. Results showed enhanced expression in exposed subjects at eight bands, namely, 6q25, 7p22, 7q22, 7q32, 13q14, 14q24, 16q22, and 16q23. Most of these bonds were fragile sites and breakpoints involved in chromosome rearrangements found in hematopoietic tumors. Moreover, six of these bands were already detected, with enhanced expression, in the first monitoring carried out on male subjects. These results indicated that fragile sites analysis is a reproducible cell response to human exposure to pesticides.

Cycling-PRINS. A method to improve the accuracy of telomeric sequence detection in mammalian chromosomes.

The presence of (TTAGGG)n telomeric sequence was investigated in a Chinese hamster tumor-derived cell line, using single primed in situ labelling (single-PRINS) and multiple cycles of amplification (cycling-PRINS). The telomeric sequence hybridized the centromere of most chromosomes, using both techniques. However, signals visible at the telomeric regions of some chromosomes and an enhancement of the frequency signals at the centromere of chromosome 1 were obtained using cycling-PRINS. These results indicate that cycling-PRINS represents a promising improvement for detection of telomeric sequence in cell lines where the conventional methods failed to demonstrate their presence.

Distribution of interstitial telomere-related sequences in the human genome and their relationship with fragile sites.

Using a computer search in the mapped human genome, we could show that interstitial telomere-related sequences are clustered in R-bands, and, in some cases, coexist with mapped fragile sites. We speculate that this association could predispose to chromosome fragility and recombination.

[Association of systemic sclerosis and psoriatic arthritis: a case report] / Associazione sclerosi sistemica ed artrite psoriasica: descrizione di un caso clinico.

The association of Systemic Sclerosis (SSc) and Psoriatic Arthritis (PsA) is unfrequent; only few cases are reported in literature. We describe a case of a patient with SSc following the onset of PsA. The disease begun with tenosynovitis, polyarthritis in association with psoriasis. After two years, Raynaud's phenomenon and sclerodactyly appeared, and, later, pulmonary interstitial fibrosis and esophageal dysfunction. The existence of a common pathogenesis of the two diseases, SSc and PsA, is discussed.

Spontaneous and aphidicolin-sensitive fragile site 3cen co-localizes with the (TTAGGG)n telomeric sequence in Chinese hamster cells.

Aphidicolin-sensitive fragile sites were analyzed in immortalized Chinese hamster embryonal fibroblast cells (CHEF18) at three different passages along their spontaneous progression toward tumorigenicity. Five fragile sites (viz., 12q22, 3cen, 3p21, 3q31, and Xq21) were detected. Three of these sites carry spontaneous aberrations and are thus regions of chromosomal instability; however, they were not involved in the formation of the clonal rearrangements that are characteristic of CHEF 18 cells. The presence of the (TTAGGG)n telomeric sequence in chromosome bands associated with fragile sites was investigated using fluorescence in situ hybridization and primed in situ labeling. A common location of fragile sites and telomeric sequence was found at the centromere of chromosome 3.

Primed in situ labeling (PRINS): a method for rapid identification and quantification of human chromosomes in both lymphocytes and sperm nuclei.

In this work, a specific primer for X alphoid satellite DNA was used to detect chromosome X through primed in situ labeling (PRINS). The method allows the rapid identification of chromosome X in metaphase and its quantification in interphase. PRINS is equally applicable to both lymphocytes and sperm nuclei.

[Unemployment and depression. Results of a psychometric evaluation]. / Inoccupazione e depressione. Risultati di una valutazione psicometrica.

The Authors evaluate the issue about the search of first employment and the depressive effects possibly linked to it. The research includes two groups of unemployed and two control groups of workers. Sex difference has been taken into account. After a preliminary semi-structured interview, all subjects have been evaluated by the means of psychological tests. The psychometric values considered include the D and Ma scales of MMPI and their difference as proposed by Diamond, and the Beck Depression Rating Scale. The results do not show a significant correlation between the search for first employment and the presence of depressive traits.

[Depressive features in unemployed individuals]. / Valenze depressive in cassintegrati.

The present research deals with a psychometric evaluation of depressive features in a group of laid-off compared with a control group of full-time workers. Sex differences has been taken into account in the comparison. (The laid-off workers belong to a group under an unemployment fund, Cassa Integrazione Guadagni, which continues paying about 80% of the ex-employee's last wage for an undetermined period of time). The psychometric elements considered include the D and Ma scales of MMPI and their difference as proposed by Diamond, and the Beck Depression Rating Scale. The results, still preliminary, show that work deprivation, even with the financial subsidy from the unemployment fund, can be considered a life event able to provoke a depressive symptomatology.

Assignment of Chinese hamster p53 gene (TP53) to chromosome band 2p31, a region not involved in the karyotypic changes of a tumorigenic cell line.

The Chinese hamster tumor-derived cell line 835T2 exhibits specific karyotypic changes, including the loss and/or translocation of genetic material. To investigate whether the p53 tumor suppressor gene was involved in the exchanges, cDNA from primary Chinese hamster cells was isolated by using sense and antisense primers of the human p53 gene. The cDNA was sequenced, and the sequence was compared with the Syrian and human p53 cDNA reported sequences. The sequence homology was very elevated, demonstrating that the cloned fragment contained part of the Chinese hamster p53 gene. The corresponding genomic fragment was also cloned and used as a biotin-labeled probe for in situ hybridization on Chinese hamster chromosome spreads. Hybridization was visualized by avidin-FITC, and the assignment was done comparing the banding obtained with BamHI restriction enzyme and the location of the fluorescent signals pattern of the same metaphase. The signals revealed that the p53 gene (TP53) is localized on Chinese hamster chromosome band 2p31, which is not involved in the karyotypic changes specific to the 835T2 cell line.

Common fragile sites on human chromosomes represent transcriptionally active regions: evidence from camptothecin.

Cellular processes involved in the expression of fragile sites (FS) have been investigated by studying the possible modulation of their induction by camptothecin, a specific topoisomerase I inhibitor. Expression of FS was induced by aphidicolin and then camptothecin was administered to cultures during G2 phase. Under these conditions, a very high number of chromosome aberrations were obtained: R-bands carrying FS were specifically involved in breakage and, in particular, the common FS (cFS) bands already expressed in aphidicolin-treated cultures were the most affected. These data show that the expressed FS are preferential targets of camptothecin, that is, regions where topoisomerase I-cleavable complexes are formed. This allows us to hypothesize that cFS could represent the cytogenetic expression of transcriptionally active regions. These treatments were able to induce, besides the known FS, four new FS, namely 1p34, 6p21, 6q25, and 15q15.

Longitudinal patterns similar to G-banding in untreated human chromosomes: evidence from atomic force microscopy.

The structure of human metaphase chromosomes, fixed according to standard procedures for optical microscopy but not treated for banding, was examined by atomic force microscopy (AFM). The images show that chromosomes display a banding pattern very similar to G-banding, detected by the AFM as a variation in the thickness of chromatin. This similarity allows the identification of individual chromosomes.

An atomic force microscope for cytological and histological investigations.

An atomic force microscope (AFM) specifically designed for cytological and histological studies and able to operate on the same scale of the highest optical magnification is described. The AFM is a non-invasive instrument; it operates on samples which do not require any kind of treatment and it can produce information that supplements and completes the information given by traditional microscopical methods. The apparatus has been used to image fixed human chromosomes and to investigate the action of trypsin during the staining for banding. First results showed that banding patterns very similar to G-banding pre-exist to staining and to trypsin treatment in human metaphase chromosomes, and that the trypsin treatment induces a structural collapse in the chromatin. The instrument was also used on thin sections of plant tissue and gave promising results. Experience confirmed that AFM is a suitable tool for this kind of investigation, and proved the importance of developing AFM microscopes specifically designed for routine use in cytology and histology, conceived for non-specialized users.