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A comprehensive study of the environmental fate of pollutants is more and more required, above all on new contaminants, i.e. pharmaceuticals. As high-resolution mass spectrometry (HRMS(n)) may be a suitable analytical approach for characterization of unknown compounds, its performance was evaluated in this study.The analyses were carried out using liquid chromatography (LC) (electrospray ionization (ESI) in positive mode) coupled with a LTQ-Orbitrap analyzer. High-resolution mass spectrometry was employed to assess the evolution of the drug transformation processes over time; accurate masses of protonated molecular ions and sequential product ions were reported with an error below 5 millimass units, which guarantee the correct assignment of their molecular formula in all cases, while their MS(2) and MS(3) spectra showed several structurally diagnostic ions that allowed characterization of the different transformation products (TPs) and to distinguish the isobaric species.The simulation of phototransformation occurring in the aquatic environment and identification of biotic and abiotic transformation products of the two pharmaceuticals were carried out in heterogeneous photocatalysis using titanium dioxide, aimed to recreate conditions similar to those found in the environmental samples. Twenty-eight main species were identified after carbamazepine transformation and twenty-nine for clarithromycin.This study demonstrates that HRMS, combined with LC, is a technique able to play a key role in the evaluation of the environmental fate of pollutants and allows elucidation of the transformation pathways followed by the two drugs.

The paper examines the transformation of phenazone (2,3-dimethyl-1-phenyl-3-pyrazolin-5-one), a widely used analgesic and antipyretic drug, under simulated solar irradiation in pure water, using titanium dioxide, and in river water. High-resolution mass spectrometry was employed to monitor the evolution of photoinduced processes. Initially, laboratory experiments were performed to simulate drug-transformation pathways in aqueous solution, using TiO(2) as photocatalyst. Thirteen main phenazone transformation products were detected, and full analysis of their MS and MS(n) spectra identified the diverse isobaric species. All these transformation products were themselves easily degraded, and no compounds were recognized to remain until 1h of irradiation. From these findings, a tentative degradation pathway is proposed to account for the photoinduced transformation of phenazone in natural waters. These simulation experiments were verified in the field, seeking phenazone in River Po water samples.

High Mobility Group I(HMGI) proteins are nuclear proteins involved in the regulation of chromatin structure and function, Elevated expression of the HMGI proteins (HMGI, HMGY and HMGI-C) has been correlated with the presence of a highly malignant phenotype in epithelial and fibroblastic rat thyroid cells, and in several experimental carcinomas, Here, we demonstrate that HMGI and HMGY proteins are expressed in human thyroid carcinomas and thyroid carcinoma cell lines, but not in adenomas, goiters, normal thyroid tissues and cells, These results indicate a correlation between HMGI and HMGY expression and the malignant phenotype of thyroid neoplasias, suggesting that these proteins may be used as markers in thyroid cancer.

BACKGROUND: Obese patients are often affected by hypertension, dyslipidaemia, impaired glucose metabolism, and suffer from cardiovascular disease (CVD), related to the characteristic metabolic alterations. AIM OF THE STUDY: To evaluate reduction of risk factors for CVDs in morbid-obese patients (body mass index (BMI)>40kg/m2) after weight loss upon bariatric surgery intervention of biliary-intestinal bypass. SUBJECTS: 45 (17 men, 28 women) morbid-obese patients (age: 19–49 y, BMI>40 kg/m2). All patients were selected on the basis of medical history, physical and biochemical evaluation and of psychiatric tests, which were performed on all individuals admitted to our Day Hospital to verify the safety of surgical intervention. MEASUREMENTS: Body weight, body composition (by dual X-ray absorptiometry, DXA), blood pressure, lipid profile, fibrinogen and glucose metabolism were monitored at baseline and 1, 3, 6, 9, 12, 24 and 36 months after surgery. RESULTS: A significant and persistent weight loss was present in all patients at the end of the 3 y follow-up period (P

The high mobility group proteins (HMGs) are a class of low molecular weight, nonhistone, nuclear proteins that bind DNA and function as transcription cofactors. This class includes the HMGI family members HMGI-C and HMGI(Y). Both are not significantly expressed in differentiated adult tissues, including fat, but their expression is induced in proliferating and transformed cells. Their involvement in the development of lipomatous tumors has been recently demonstrated for HMGI-C, which is encoded by a gene located at 12q15, the chromosomal segment often rearranged in ordinary lipomas. The same chromosomal segment is consistently amplified in the ring and giant marker chromosomes of atypical lipomatous tumors (ALTs), a term used to designate tumors previously labeled as well differentiated liposarcomas or atypical lipomas. The involvement of HMGI(Y) is strongly suspected as the gene coding for HMGI(Y) is located at 6p21, a chromosomal segment rearranged in a subset of ordinary lipomas. HMGI-C or HMGI(Y) protein expression was analyzed immunohistochemically in a group of 39 well differentiated adipose neoplasms (19 lipomas and 20 ALTs) of known karyotype using polyclonal antibodies raised against a recombinant protein (HMGI-C) and against a synthetic peptide (HMGI(Y)). The results of this study demonstrate that HMGI proteins are commonly expressed in well differentiated adipose neoplasms. Seventeen of twenty ALTS (85.0%), all of which had ring or giant marker chromosomes with amplification of 12q13-15, strongly expressed HMGI-C. HMGI-C expression was detected in 7 of 11 ordinary lipomas (63.6%) with alterations at 12q14-15 and in one case with an abnormal karyotype that included double minute chromosomes. HMGI-C immunoreactivity correlates with 12q13-15 chromosomal alterations (P = 0.001). HMGI(Y) reactivity was demonstrated in only two ordinary lipomas: one with 6p21 rearrangement and one with normal karyotype. No significant HMGI(Y) expression was found in the ALT group. The finding of aberrant expression of HMGI proteins in well differentiated adipose neoplasms in association with 12q13-15 and 6p21 chromosomal changes supports the proposed pathogenetic role of this group of proteins in the development of adipose tissue tumors.

22035 Background: Preclinical data suggest that increased expression of high mobility group A2 (HMGA2) protein could enhance chemosensitivity towards doxorubicin in breast cancer cells. The present...

A method is presented for sensitive staining of the HMG14 and 17 proteins in polycrylamide gels pre-stained with Coomassie Blue R250. The procedure involves binding negatively and positively charged polycyclic aromatic compounds to the proteins followed by staining with silver using the method of Wray et al. (1981)[4].

The conformational behaviour of poly( l -tyrosine) has been studied in dimethylsulphoxide/water and dimethylsulphoxide/trimethylphosphate solvent systems. The evidence provided by nuclear magnetic resonance and infrared spectroscopy shows that the conformation of the polymer in dimethylsulphoxide is most probably the random coil. Addition of 20% D 2 O or 50% trimethylphosphate to the dimethylsulphoxide/poly( l -tyrosine) solution results in a conformational transition of the polymer which can be followed by both infra-red and nuclear magnetic resonance spectroscopy and changes in optical rotation. The evidence supports the conclusion that this transition is to a largely, though not fully, right-handed α-helical conformation.

A study has been made of the reversible solid state transitions in a series of eleven zinc(II) di-n-alkylphosphinate polymers, [Zn(R2PO2)2]n, with R from C4 to C18, by means of differential scanning calorimetry and x-ray analysis. Wide line n.m.r. measurements have been also made for some compounds. The existence of two crystal forms, denoted α and β, has been established for the compounds examined. The results indicate that the α→β polymorphic transition is associated with disordering of the hydrocatbon side chains.

Lymphocyte dialysates obtained from healthy volunteers and leukemic subject were chromatographed on Bio-Gel P-4. Partially purified fractions were assayed for neutrophil locomotion: a positive chemokinetic effect and no chemotactic activity was observed in both samples.

The results of an investigation on the interaction of proflavine and of ethidium bromide with DNA (calf thymus) in dilute aqueous solution are reported. The binding of the two dyes by DNA has been studied by means of microcalorimetric and of equilibrium dialysis measurements. Data on the thermodynamics of dimerization of both proflavine and ethidium bromide in aqueous solution obtained on the basis of spectroscopic and/or calorimetric experiments are also reported. The enthalpy data show that dye-dimerization and dye "strong" interaction with DNA are energetically favourable and quite similar while only in the latter case the phenomenon is also entropy driven. This is taken as further evidence in support of the concept that "strong" interaction-of both proflavine and ethidium bromide with DNA means dye molecules intercalation into the native, double helical structure of the biopolymer.

Résumé. — Dans cet article nous avons essayé de présenter les éléments essentiels de la culture de Cris de Roumanie, qui constitue le Néolithique le plus ancien du Bas-Danube. Les thèses principales soutenues dans l'article sont au nombre de quatre. 1) Le synchronisme de Starcevo (en Yougoslavie), Koros (en Hongrie) de la Céramique Linéaire (en Tchécoslovaquie), ainsi que de Karanovo I et Kremicovci en Bulgarie avec Cris. 2) Le refus d'une division des cultures de Starcevo et de Cris en quatre phases. 3) Le développement indépendant de toutes les cultures du Néolithique ancien de Roumanie, sans relations notables avec le Proche-Orient et la Méditerranée Occidentale. 4) Le postulat d'une continuité Cris-Vinca-Turdas et Petresti., Giancotti-Tassone Vicente. Observations sur quelques aspects de la culture de Cris du Néolithique ancien de la Roumanie. In: Bulletin de la Société préhistorique française. Études et travaux, tome 67, n°1, 1970. pp. 323-334.

Nuclei isolated from the liver of rats undergoing an acute inflammatory reaction induced by turpentine treatment show increased RNA synthesis. This increase is essentially determined by a faster polyribonucleotide-elongation rate while the number of transcribing polymerase molecules is unchanged. The sensitivity of chromatin to micrococcal-nuclease digestion and the composition of chromosomal proteins are not affected by the acute-phase process. Therefore the increased RNA synthesis by liver nuclei from acutely inflamed rats does not seem to correlate with major changes in chromatin structure.

The mixed-ligand species Zn[(R12PO2)2 – 2x(R22PO2)2x](R1= Bun; R2= n-C6H13 or n-C10H21) are linear coordination copolymers with a random distribution of two different bridging phosphinate groups along the molecular chains. Single crystals of the copolymer {Zn[Bun2PO2(n-C6H13)2PO2]}n are disordered. A three-dimensional X-ray analysis has been attempted. The crystals contain chains characterised by a backbone structure with alternate single and triple bridging phosphinate groups between tetrahedral metal atoms. The structural features common to those of the β-form of zinc(II) di-n-butylphosphinate polymer are discussed.

The characterisation of beryllium phosphinates of stoicheiometry Be(Bun2PO2)2, Be(Ph2PO2)2, and of some mixed-ligand species Be[(Bun2PO2)2 – 2x(Ph2PO2)2x] is reported. The compounds richer in alkyl groups give stable solutions in hydrocarbon and halogenated solvents. Solution properties indicate that these compounds are linear polymers which undergo a reversible degradation influenced by the nature of the solvent, concentration, and temperature. The mixed-ligand species can be crystallised and are formulated as random copolymers. Isomorphous replacement of the two kinds of phosphinate groups within the crystal lattice of the related homopolymers is observed. The X-ray diffraction fibre photographs indicate that the homopolymer [Be(Bun2PO2)2]n and the copolymers {Be[(Ben2PO2)2 – 2x(Ph2PO2)2x]}n(x= 0·20 or 0·33) have a backbone structure with alternate single and triple bridging phosphinate groups between tetrahedral metal atoms. The same chain-type structure is proposed for the homopolymer [Be(Ph2PO2)2]n and the copolymers richer in diphenylphosphinate groups.

A study has been made of the iso- and poly-morphism phenomena in a series of mixed-ligand species, {M[(Bun2PO2)2 – 2x(Ph2PO2)2x]}n(M = ZnII or CoII), by means of differential scanning calorimetry and X-ray analysis. These compounds are linear copolymers and the di-n-butylphosphinate and diphenylphosphinate groups are randomly distributed along the molecular chains. The homopolymer, [Zn(BunPhPO2)2]n, has been studied as a model system with a disordered positioning of the n-butylphenylphosphinate groups along the molecular chains. This compound exhibits polymorphism and its γ-form is isomorphous with that of [Zn(Ph2PO2)2]n.The zinc(II) mixed-ligand species are crystalline and isomorphous with the blue cobalt(II) compounds. Their solubility in non-co-ordinating solvents decreases with increasing the content of Ph2PO2– groups. Isomorphous substitution of some Ph2PO2– groups with Bun2PO2– groups in the crystal lattice of the [Zn(Ph2PO2)2]n homopolymer or of some Bun2PO2– groups with Ph2PO2– groups in the crystal lattice of the [Zn(Bun2PO2)2]n homopolymer takes place. On the basis of the structures of the crystalline form of [Zn(Bun2PO2)2]n, stable at room temperature, and of the γ-form of [Zn(BunPhPO2)2]n, the solid-state transitions and the isomorphism and polymorphism phenomena of the various co-ordination polymers are discussed.

High mobility group A 1 (HMGA1) proteins are chromatinic factors, which are absent or expressed at very low levels in normal adult tissues, while they are over-expressed in several human malignant tumors. In this study, HMGA1 protein expression was investigated by immunohistochemistry in a series of 44 epithelial ovarian specimens, which included four normal ovarian tissues, 29 primary invasive carcinomas, one metastatic ovarian tumor and 10 low malignant potential (LMP) tumors. HMGA1 staining was not detected in normal ovarian surface epithelium, which is the area from which ovarian adenocarcinoma frequently arises. HMGA1 proteins were expressed at low levels in some LMP tumors, whereas they were present in abundance in most of the primary ovarian adenocarcinomas. RT-PCR and western blot analysis correlated with immunohistochemical data. We demonstrated that the suppression of HMGA1 protein synthesis by an adenovirus carrying the HMGA1 gene in antisense orientation (Ad-Yas-GFP) inhibited the growth of two human ovarian carcinoma cell lines (OVCAR-5 and OVCAR-8). These results confirm HMGA1 over-expression as a general feature of human malignant neoplasias, including ovarian cancer and suggest that suppression of HMGA1 protein synthesis by an antisense adenoviral vector may represent a new and promising gene therapy for the treatment of ovarian cancer.

Elevated expression of HMGA1 and HMGA2 proteins is correlated with a highly malignant phenotype in several human tumors. We previously demonstrated that the block of HMGA2 protein synthesis prevented rat thyroid cell transformation by murine retroviruses. Suppression of HMGA2 synthesis was associated with lack of induction of HMGA1 proteins suggesting that both HMGA1 and HMGA2 play a role in the process of neoplastic transformation. To determine the role of the HMGA1 gene in thyroid cell transformation, we blocked HMGA1 protein synthesis by an antisense methodology. Here we report that transfection of an HMGA1 cDNA antisense construct into a normal rat thyroid cell line (FRTL-5 Cl2), followed by infection with Kirsten murine sarcoma virus (KiMSV), generated a transformed cell line that expresses high levels of the v-ras-Ki oncogene and that does not require thyroid-stimulating hormones for growth. However, this cell line does not show the malignant phenotype, i.e., it neither grows in soft agar nor induces tumors after injection in athymic mice. Moreover, the lack of the neoplastic phenotype in the virus-infected thyroid cells carrying the HMGA1 antisense construct correlates with the absence of induction of AP-1 transcriptional activity.

HMGI(Y) proteins are overexpressed in experimental and human malignancies, including colon, prostate and thyroid carcinomas. To determine at which step of the carcinogenic process HMGI(Y) induction occurs, we analysed the expression of the HMGI(Y) proteins in hyperplastic, preneoplastic and neoplastic tissues of colorectal origin by immunohistochemistry. All the colorectal carcinomas were HMGI(Y)-positive, whereas no expression was detected in normal colon mucosa tissue. HMGI(Y) expression in adenomas was closely correlated with the degree of cellular atypia. Only 2 of the 18 non-neoplastic polyps tested were HMGI(Y)-positive. These data indicate that HMGI(Y) protein induction is associated with the early stages of neoplastic transformation of colon cells and only rarely with colon cell hyperproliferation.

The HMGA family is comprised of four proteins: HMGA1a, HMGA1b, HMGA1c and HMGA2. The first three proteins are products of the same gene, HMGA1, generated through an alternative splicing mechanism. The HMGA proteins are involved in the regulation of chromatin structure and HMGA DNA-binding sites have been identified in functional regions of many gene promoters. Rearrangements of the HMGA2 gene have been frequently detected in human benign tumors of mesenchymal origin including lipomas. 12q13-15 chromosomal translocations involving the HMGA2 gene locus, account for these rearrangements. The HMGA proteins have three AT-hook domains and an acidic C-terminal tail. The HMGA2 modifications consist in the loss of the C-terminal tail and fusion with ectopic sequences. A pivotal role of the HMGA2 rearrangements in the process of lipomagenesis is suggested by experiments showing that transgenic mice carrying a truncated HMGA2 gene showed a giant phenotype together with abdominal/pelvic lipomatosis. As HMGA2 null mice showed a great reduction in fat tissue, a positive role of the HMGA2 gene in adipocytic cell proliferation is proposed. More recently, similar alterations of the HMGA1 gene have been described. As the block of the HMGA1 protein synthesis induces an increase in growth rate of the pre-adipocytic cell line 3T3-L1, we suggest a negative role of the HMGA1 proteins in adipocytic cell growth and, therefore, we propose that adipocytic cell growth derives from the balance of the HMGA1 and HMGA2 protein functions.

HMGI-C is a nuclear architectural factor which is expressed during embryogenesis but not in adult tissues while it becomes re-expressed following neoplastic transformation. In this paper we identify the promoter region of the mouse Hmgi-c gene and by stepwise deletion of the 5' sequences we map the promoter activity of the most abundant transcript to a very short fragment containing a long polypyrimidine/polypurine (ppyr/ppur) tract. We demonstrate that this tract is a multiple binding site for the transcription factors Sp1 and Sp3 and that in Drosophila SL2 cells, Spl activates the Hmgi-c promoter. In addition, another transcription factor, CTF/NF-1, binds the proximal promoter immediately downstream of this region and its mutation decreases transcription in NIH-3T3 cells. This study identifies factors responsible for the basal activity of Hmgi-c gene and provides a foundation for further analysis of the mechanism of its regulation.

A correlation has previously been demonstrated between the presence of the three HMGI proteins (HMGI, HMGY, and HMGI-C) and the expression of a highly malignant phenotype in epithelial and fibroblastic rat thyroid cells; this being subsequently extended to experimental thyroid, lung, prostate, mammary, and skin carcinomas. Recently, we have demonstrated that expression of HMGI and HMGY proteins, coded for by the HMGI(Y) gene, is associated with the malignant phenotype of human thyroid neoplasias. Here, we show that HMGI(Y) gene expression is present both at the RNA and protein level in human colorectal carcinoma cell lines and tissues examined in this study. Conversely, no HMGI(Y) proteins were detected in normal intestinal mucosa. Therefore, these results suggest an involvement of HMGI and HMGY proteins overexpression in colorectal tumorigenesis.

Four malignant tumors induced in mouse by different experimental procedures were compared as regards their high-mobility-group (HMG) proteins. All tumors showed the complete set of three HMG proteins which we call HMGI-C, I-D, and I-E. The presence of the three HMGI proteins is a characteristic of the transformed phenotype regardless of whether the tumor was chemically, virally, or spontaneously derived. However, the level of expression of the HMGI proteins is not constant in the four tumors. Using reverse-phase HPLC, the individual HMGI proteins were isolated from the spontaneously derived tumor (Lewis lung carcinoma) and shown by amino acid analysis to be similar to those previously obtained from a tumor grown in nude mice by inoculation of in vitro-transformed cells.

The HMGI protein family includes three proteins, named HMG-I, HMG-Y and HMGI-C. The first two proteins are coded for by the same gene, HMGI (Y), through an alternative splicing mechanism. Their expression is elevated in neoplastic tissues and cells and this overexpression has a causal role in the process of cellular neoplastic transformation. We demonstrate that the HMGI (Y) gene is expressed at very low levels in normal adult tissues, whereas in embryonic tissues it is expressed at high levels comparable to those detected in neoplastic tissues. Specifically, a very high expression of the HMGI (Y) gene was detected in all embryonic tissues at 8.5 dpc. Then in the following days, even though the gene is expressed essentially in all tissues, an abundant gene expression was restricted to some tissues. These results indicate an important role of the HMGI (Y) gene in development.

The expression of nuclear proteins high mobility group (HMG) I and HMGY was investigated in intraepithelial and invasive lesions of the uterine cervix. Human carcinoma cell lines C-41, ME-180, and CaSki were used for testing protein expression in neoplastic cells from the cervix. Morphological grading of the dysplasias (CIN 1, CIN 2, and CIN 3) and invasive carcinomas from formalin-fixed paraffin-embedded samples parallels the degree of nuclear immunostaining obtained using a polyclonal antibody raised against the amino-terminal region of HMGI(Y) proteins. The immunostaining obtained with HMGI(Y) antibody was compared with that observed using the antibody Ki-67, and the results were similar. We suggest the use of HMGI(Y) antibody in clinical oncology as a useful marker of intraepithelial lesions and invasive carcinomas.

The high mobility group proteins (HMGs) are a class of low molecular weight, nonhistone, nuclear proteins that bind DNA and function as transcription cofactors. This class includes the HMGI family members HMGI-C and HMGI(Y). Both are not significantly expressed in differentiated adult tissues, including fat, but their expression is induced in proliferating and transformed cells. Their involvement in the development of lipomatous tumors has been recently demonstrated for HMGI-C, which is encoded by a gene located at 12q15, the chromosomal segment often rearranged ill ordinary lipomas. The same chromosomal segment is consistently amplified in the ring and giant marker chromosomes of atypical lipomatous tumors (ALTs), a term used to designate tumors previously labeled as well differentiated liposarcomas or atypical lipomas. The involvement of HMGI(Y) is strongly suspected as the gene coding for HMGI(Y) is located at 6p21, a chromosomal segment rearranged in a subset of ordinary lipomas. HMGI-C or HMGI(Y) protein expression was analyzed immunohistochemically in a group of 39 well differentiated adipose neoplasms (19 lipomas and 20 ALTs) of known karyotype using polyclonal antibodies raised against a recombinant protein (HMGI-C) and against a synthetic peptide (HMGI(Y)) Tbe results of this study demonstrate that HMGI proteins are commonly expressed in well differentiated adipose neoplasms. Seventeen of twenty ALTS (85.0%), all of which had ring or giant marker chromosomes with amplification of 12q13-15, strongly expressed HMGI-C. HMGI-C expression was detected in 7 of 11 ordinary lipomas (63.6%) with alterations at 12q14-15 and in one case with an abnormal karyotype that included double minute chromosomes. HMGI-C immunoreactivity correlates with 12q13-15 chromosomal alterations (P = 0.001). HMGI(Y) reactivity was demonstrated in only two ordinary lipomas: one with 6p21 rearrangement and one with normal karyotype. No significant HMGI(Y) expression was found in the ALT group. The finding of aberrant expression of HMGI proteins in well differentiated adipose neoplasms ill association with 12q13-15 and 6p21 chromosomal changes supports the proposed pathogenetic role of this group of proteins in the development of adipose tissue tumors.