Evidence for the presence in calf thymus of a peptidic factor controlling DNA transcription in vitro.

A thymic factor causes a strong inhibition of the DNA-directed RNA polymerase reaction in vitro. The active factor was isolated from aqueous ultrafiltered thymus extracts and purified by means of chromatography on DEAE-cellulose and then on Dowex 50 WX2. The purified thymic factor was characterized as a peptide of low molecular weight (less than 5000). The biological activity of the thymic factor cannot be attributed to the presence of a nuclease or of a histone fragment. The RNA synthesis is controlled by this factor by means of electrostatic interactions between the peptide compound and DNA. Inhibitory activity on RNA synthesis was absent from kidney extracts.

Detection of functional platelet-activating factor receptors on leukemic B cells of chronic lymphocytic leukemic patients.

Although platelet-activating factor receptors (PAF-R) are reported on normal B cells, few results are available concerning leukemic ones. We demonstrated functional PAF-R on cell and nuclear surfaces of leukemic B cells of chronic lymphocytic leukemic (CLL) patients. Analysis of 102 patients revealed dramatic differences for their membrane PAF-R expression, a result that might be related to their plasma IL-4 levels. In the light of the potent immunoregulatory role of PAF on B cell physiology, it is suggested that the presence or absence of PAF-R on leukemic B cells may profoundly affect their in vivo behavior.

Special care units for demented patients: a multicenter study.

The local government of Regione Lombardia, Italy, recently (1994) funded a clinical and research project specifically devoted to dementia (Piano Alzheimer). A central role in this project has been reserved for the special care units (SCUs) for demented patients with behavioral disturbances. In order to evaluate their effectiveness, eight SCUs took part in this study. A specifically designed care program, focusing on environment and staff, was implemented in each SCU. Cognitive, functional, and somatic health status, and use of psychotropic drugs and of physical restraints were assessed at baseline, and after 3 and 6 months in 55 consecutively admitted patients. The data show an overall reduction in behavioral disturbances and a decreased use of psychotropic drugs and physical restraints.

[1000 osteoarthrosis patients in treatment with GGGs: an assessment of joint pain]. / 1000 pazienti osteoartrosici in trattamento con GGGs: valutazione del dolore articolare.

The analgesic activity of a chondroprotective agent--galactosamineglucuronoglycan sulphate (Condral) was evaluated in the course of an open investigation in a large number of outpatients attending the Istituto Emilio Bruzzone of Genoa or related Centres. Out of 1720 outpatients seen, 1000 were treated for 2 years with 800-1200 mg daily of GGG in one single administration (2-3 sachets or capsules of 400 mg) By the results obtained GGG was shown to have analgesic activity in a large number of cases. Tolerance of the treatment was good and no significant clinical side effects were ever observed.

Testing the effect of selectors in the control of bulking and foaming in full scale activated-sludge plants.

Selectors were operated at four full-scale activated sludge plants to control bulking and foaming problems due to filamentous microorganisms. Selector effectiveness was not related to reduction of biodegradable organic matter in the contact zone, but was related to soluble COD levels in selector effluent. Significant reductions in the numbers of filamentous m icroorganisms were reported.

[The body and its meanings]. / Il corpo e la corporeità. Riflessione dovuta per un operatore sanitario.

The human body is the natural object of studies, analyses and care in the health field. The Authors, starting from this remark and taking the body language and its meaning into consideration, point out that the body contact allows either to deliver an adequate nursing care or to take off dignity and humanity. A concise review of the human body significance during the course of history makes us aware that, as a matter of fact, the different and opposite conceptions that followed one another haven't made the concept of body definitively clear. Inside the care field, the knowledge and use of the body language--complementary to the verbal one--assumes many different meanings, that can condition the relation process in a substantial way.

MYCN gene expression is required for the onset of the differentiation programme in neuroblastoma cells.

Neuroblastoma is an embryonic tumour of the sympathetic nervous system and is one of the most common cancers in childhood. A high differentiation stage has been associated with a favourable outcome; however, the mechanisms governing neuroblastoma cell differentiation are not completely understood. The MYCN gene is considered the hallmark of neuroblastoma. Even though it has been reported that MYCN has a role during embryonic development, it is needed its decrease so that differentiation can be completed. We aimed to better define the role of MYCN in the differentiation processes, particularly during the early stages. Considering the ability of MYCN to regulate non-coding RNAs, our hypothesis was that N-Myc protein might be necessary to activate differentiation (mimicking embryonic development events) by regulating miRNAs critical for this process. We show that MYCN expression increased in embryonic cortical neural precursor cells at an early stage after differentiation induction. To investigate our hypothesis, we used human neuroblastoma cell lines. In LAN-5 neuroblastoma cells, MYCN was upregulated after 2 days of differentiation induction before its expected downregulation. Positive modulation of various differentiation markers was associated with the increased MYCN expression. Similarly, MYCN silencing inhibited such differentiation, leading to negative modulation of various differentiation markers. Furthermore, MYCN gene overexpression in the poorly differentiating neuroblastoma cell line SK-N-AS restored the ability of such cells to differentiate. We identified three key miRNAs, which could regulate the onset of differentiation programme in the neuroblastoma cells in which we modulated MYCN. Interestingly, these effects were accompanied by changes in the apoptotic compartment evaluated both as expression of apoptosis-related genes and as fraction of apoptotic cells. Therefore, our idea is that MYCN is necessary during the activation of neuroblastoma differentiation to induce apoptosis in cells that are not committed to differentiate.

Drug hypersensitivity: epidemiology and risk factors.

Drug allergies are heterogeneous and multifactorial diseases and are always the consequence of an exaggerated immune-mediated reaction. Previously described models of immunologic mechanisms (mainly based on Gell and Combs' classification) cannot fully explain the physiopathology of these diseases; it seems therefore important to identify risk factors. Clinical and biologic tests are helpful diagnostic tools but are limited in their sensitivity and reliability and are certainly not predictive. Epidemiologic data supply information concerning the prevalence of drug hypersensitivity: female gender, concomitant infections (HIV, herpes) and concurrent illnesses (systemic lupus erythematosus) are all significant risk factors. Another host-related factor is the genetic predisposition of patients and is currently under investigation in our laboratory. Most genetic studies concern HLA haplotype association or polymorphism in genes encoding drug-metabolising enzymes. A current study by our group seems to implicate polymorphisms within the promoter of IL-10, a cytokine with anti-inflammatory properties. The chemical properties of the drug and the treatment regimen also influence the development of drug allergies.

IL-10 promoter and IL4-Ralpha gene SNPs are associated with immediate beta-lactam allergy in atopic women.

BACKGROUND: Allergic reactions to beta-lactam antibiotics represent the most frequent cause of immunological drug reactions. OBJECTIVE: This study evaluates the involvement of genetic susceptibility factors in patients with immediate allergic reactions to beta-lactams. We examined 15 single nucleotide polymorphisms (SNP) of genes coding proteins implicated in immunoglobulin (Ig)E synthesis regulation. METHODS: We performed a case-control study involving 44 patients with immediate beta-lactam allergy and 44 control subjects, all matched for sex and atopy. Interleukin (IL)-4, IL-13, IL-4Ralpha, signal transducer and activator of transcription 6 (STAT6), interferon (IFN)-gammaR1, IFN-gammaR2 and FcepsilonRIbeta gene polymorphisms were determined using polymerase chain reaction (PCR) restriction fragment length polymorphism, and IL-21R gene and IL-10 promoter polymorphisms by direct sequencing. RESULTS: Our analysis did not reveal differences in the distribution of the 15 SNPs between allergic patients and controls. However, among atopic subjects, we found two distinct significant associations between immediate beta-lactam allergy in women and the Ile75Val variant of IL-4Ralpha gene (P = 0.012, OR = 5.4, CI: 1.16-27.7), and two linked IL-10 promoter gene polymorphisms, -819C>T and -592 C>A (P = 0.023, OR = 17.5, CI: 1.26-533.07). In contrast, we observed no association in allergic male subjects in the atopic population. Interestingly, the IL-4Ralpha Ile75Val variant could have a paradoxal protective effect in atopic male patients (P = 0.004, OR = 0.07, CI: 0.01-0.66). CONCLUSION: Our findings suggest that polymorphisms in the IL-10 promoter and IL-4Ralpha genes are genetic factors that favour beta-lactam immediate allergies in female patients with atopy.

Amplification of template capacity of mammalian DNA following extraction of low molecular weight peptides.

Experiments were carried out to evaluate the physical and functional modifications of DNA from several mammalian tissues following alkaline extraction (pH 9.5). The results indicate that such extraction provokes removal of firmly bound low molecular weight non-histone peptides involved in a general mechanism regulating the template capacity of DNA.

Stabilization of double-stranded DNA molecule by non-histone peptidic effector from calf thymus.

A peptidic effector from calf thymus causes a strong stabilization of DNA double-stranded molecule in vitro. The active factor was isolated from aqueous ultrafiltered thymus extracts and purified by means of chromatography on DEAE-cellulose and then on Dowex 50 WX2. The purified thymic factor was characterized as a peptide of low molecular weight (less than 5000). The biological activity of the thymic factor cannot be attributed to a histone fragment. Melting data of the control DNA and of the DNA-active factor complex in various conditions of ionic strength and dielectric constant of the solution medium are recorded.

Behaviour in biotin-deficient rats of thymic peptides controlling DNA transcription.

Biotin-deficient rats show a slowing down of the growth and an involution of the thymus. The amount of the thymic peptides controlling DNA template, if referred to the thymus weight is higher in deficient than in control rats; no significant difference is noticed among the contents of the active peptides when evaluated per rat. The inhibiting activity on RNA synthesis is the same for the peptides extracted from normal and from biotin-deficient rat thymus.

Low molecular weight peptides controlling transcription are present in the calf thymus chromatin structure.

A calf thymus peptide fraction controlling DNA and chromatin template has been purified by DNA-cellulose and Dowex 50 WX2 chromatography and its amino acid composition determined. The active peptide fraction can be extracted in high pH buffer from calf thymus native chromatin previously deproteinized by chloroform-isamyl alcohol and phenol. These data demonstrate that the thymic peptide(s) is (are) a chromatin protein constituent strongly linked to DNA. The specificity in association of the peptide(s) to DNA has also been considered.

Inhibition of exogenous mRNA translation in a cell-free system by chromatin peptides from calf thymus.

Low molecular weight chromatin peptides isolated from calf thymus by affinity chromatography on DNA-cellulose inhibit significantly translation of exogenous isolated mRNA in a reticulocyte cell-free system. Translation with endogenous mRNA present in the system is not inhibited by low peptide concentrations. The data obtained combined with the previous findings suggest that chromatin peptides control gene expression at two levels: transcription and translation.

Specific thymic peptides-DNA interaction. Correlation with the possible stereochemical kinking scheme of DNA.

Low molecular weight peptides from calf thymus cause a strong dose-dependent stabilization of the DNA. The strength od DNA-peptide interaction is pH-dependent and decreases repidly above pH 6.5. Moreover the complete kinetics of DNA denaturation and renaturation demonstrates that the peptide fraction increases significantly the DNA renaturation mostly at low temperature, showing that the interaction DNA-thymic effector helps the recombination of complementary DNA segments. The DNA stabilization rate by the peptide fraction is comparable to that obtained by means of high concentration of histones or synthetic polycationic peptides. However, the lack of basic amino acids in the peptide structure is not in favor of strong electrostatic interactions and implies a specific binding of peptide to DNA. The possible correlation of the specific thymic peptides-DNA interaction with the stereochemical kinking scheme of DNA is discussed.

Mouse embryonic stem cell sorting for the generation of transgenic mice by sedimentation field-flow fractionation.

Mouse embryonic stem (ES) cells are an important tool for generation of transgenic mice and genetically modified mice. A rapid and efficient separation of ES cells that respects cell integrity, viability, and their developmental potential while also allowing purified ES fraction collection under sterile conditions might be of great interest to facilitate the generation of chimeric animals. In this study, we demonstrated for the first time the effectiveness of a sedimentation field-flow fractionation (SdFFF) cell sorter to provide, with a characteristic DNA content, a purified ES cell fraction and with a high in vivo developmental potential to prepare transgenic mice by generation of chimeras having a high percentage of chimerism.

[The organization of a working group for the prevention of decubitus lesions]. / Organizzazione di un gruppo di lavoro per la prevenzione delle lesioni da decubito.

The authors report the results of a two years monitoring program focused on patients at risk of and with bedsores in the general hospital of Castelfranco Veneto. The purpose of the multidisciplinary exercise (which saw the participation of nurses, pharmacists, doctors) was the activation and the evaluation of treatment protocols. The overall recruited sample included 679 patients, with a prevalence of 61% of lesions at the occasion of the first observation, and of 57% in the last one. The hardly quantitatively different prevalence is better understood from a qualitative point of view: the number of lesions per patients drops slightly from 1.9 to 1.7; one out of four of the "at risk" patients became "at no risk"; a 30% improvement is observed with respect to treatment efficacy. It is suggested that a positive effect may be obtained through a sustained attention to the problem and the careful definition of the interventions to be planned.