A new silanizing agent tailored to surface bio-functionalization.

Amino-terminated surfaces can be effectively obtained by means of silanizing agents, realizing surfaces suitable for the purification of biomarkers of several pathologies. Since the level of biomarkers, such as microRNAs and cell-free DNA, into circulation may be extremely low, new and ameliorated capturing molecules and protocols are highly required. In this work, a new silane, acetone-imine propyl trimethoxysilane (AIPTMS), is synthesized with a simple and elegant reaction, via the nucleophilic addition of the primary amino group to the carbonyl group of acetone. AIPTMS and APTMS were used to silanize silicon oxide surfaces, which were characterized chemically (XPS) and morphologically (AFM). The two types of surfaces were chemically similar, but behaved very differently both for surface morphology and functional properties. The AIPTMS-modified surface was indeed very smooth and homogeneous with respect to the APTMS-modified surface. Moreover, the AIPTMS surface captured larger amounts of nucleic acids almost immediately after preparation, while APTMS-based functional surfaces needed longer time to reach comparable efficiency. AIPTMS shows several advantages over standard aminosilanes, as it realizes a more homogeneous surface coverage that, in turn, produces an improved response towards the capture of nucleic acids. AIPTMS is a very promising reagent for the reliable and reproducible preparation of active biofunctional surfaces for the purification and analysis of circulating biomarkers.

Long-lasting genomic instability following arsenite exposure in mammalian cells: the role of reactive oxygen species.

Previously, we reported that the progeny of mammalian cells, which has been exposed to sodium arsenite for two cell cycles, exhibited chromosomal instability and concurrent DNA hypomethylation, when they were subsequently investigated after two months of subculturing (about 120 cell generations) in arsenite-free medium. In this work, we continued our investigations of the long-lasting arsenite-induced genomic instability by analyzing additional endpoints at several time points during the cell expanded growth. In addition to the progressive increase of aneuploid cells, we also noted micronucleated and multinucleated cells that continued to accumulate up to the 50th cell generation, as well as dicentric chromosomes and/or telomeric associations and other complex chromosome rearrangements that began to appear much later, at the 90th cell generation following arsenite exposure. The increasing genomic instability was further characterized by an increased frequency of spontaneous mutations. Furthermore, the long-lasting genomic instability was related to elevated levels of reactive oxygen species (ROS), which at the 50th cell generation appeared higher than in stable parental cells. To gain additional insight into the continuing genomic instability, we examined several individual clones isolated at different time points from the growing cell population. Chromosomally and morphologically unstable cell clones, the number of which increased with the expanded growth, were also present at early phases of growth without arsenite. All genomically unstable clones exhibited higher ROS levels than untreated cells suggesting that oxidative stress is an important factor for the progression of genomic instability induced by arsenite.

Estimation of fetal weight by measurement of fetal thigh soft-tissue thickness in the late third trimester.

OBJECTIVE: The accuracy of current formulae for the sonographic estimation of fetal weight (EFW) is compromised by significant intra- and interobserver variability of biometrical measurements, particularly circumferences. The aim of this study was to assess the reliability of the linear measurement of mid-thigh soft-tissue thickness (STT) and to derive a novel formula for EFW. METHODS: This was a prospective study involving 388 singleton uncomplicated pregnancies. There were three consecutive phases: (1) to verify the relationship between STT and birth weight, (2) to derive a novel formula for EFW using femur length and STT only, and (3) to test the accuracy of the new equation. Only the 290 patients who delivered within 48 h of measurement were considered for the analysis. A comparison with other formulae was performed. RESULTS: STT was significantly correlated with both abdominal circumference and birth weight (r(2) = 0.36 and 0.46, respectively; P < 0.001). Both intra- and interobserver variability were satisfactory (0.44 +/- 0.27 and 0.57 +/- 0.35 mm, respectively). The equation for EFW was developed using multiple stepwise regression analysis (EFW = - 1687.47 + (54.1 x femur length) + (76.68 x STT)) and tested prospectively on 69 patients. The new formula yielded results (r = 0.79) that were slightly better in accuracy than two other published equations, and had an absolute mean error of < 15% in 97% of cases. CONCLUSIONS: Our findings confirm the potential of the linear measurement of mid-thigh STT as a valuable parameter for the sonographic assessment of fetal growth and EFW. Our new equation is apparently at least as reliable as the most widely used formulae for EFW.

Bradyrhizobium sp. nodulating the Mediterranean shrub Spanish broom (Spartium junceum L.).

AIMS: The molecular diversity of 25 strains of rhizobia, isolated in Sicily from root nodules of the Mediterranean shrubby legume Spanish broom (Spartium junceum L.), is presented in relation to the known rhizobial reference strains. METHODS AND RESULTS: Our approach to the study of the S. junceum rhizobial diversity combined the information given by the 16S and the intergenic spacer (IGS) 16S-23S rDNA polymorphic region by obtaining them in a single polymerase chain reaction (PCR) step. The PCR fragment size of the S. junceum isolates was 2400-2500 bp and that of the reference strains varied from 2400 in Bradyrhizobium strains to 2800 in Sinorhizobium strains. Inter- and intrageneric length variability was found among the reference strains. Restriction fragment length polymorphisms (RFLP) analysis allowed us to identify eight genotypes among the S. junceum rhizobia that were clustered into two groups, both related to the Bradyrhizobium lineage. Sequencing of representative strains of the two clusters confirmed these data. The 16S-IGS PCR-RFLP approach, when applied to rhizobial reference strains, allowed very close species (i.e. Rhizobium leguminosarum/R. tropici) to be separated with any of the three enzymes used; however, cluster analysis revealed inconsistencies with the 16S-based phylogenesis of rhizobia. CONCLUSIONS: Rhizobia nodulating S. junceum in the Mediterranean region belong to the Bradyrhizobium lineage. Our results confirm the resolution power of the 16S-23S rDNA in distinguishing among rhizobia genera and species, as well as the usefulness of the PCR-RFLP method applied to the entire 16S-IGS region for a rapid tracking of the known relatives of new isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: The present paper is, to our knowledge, the first report on rhizobia nodulating a Mediterranean wild woody legume.

Malignant mixed mullerian tumor of the ovary: report of four cases.

INTRODUCTION: Malignant mixed mullerian tumor (MMMT) of the ovary is an extremely rare gynaecologic neoplasm that represents 1% of the malignances of this organ. Stage I disease is rare because it is asymptomatic in early stage. We describe four cases. CASE REPORTS: In the Department of Obstetrics and Gynecology of the University of Bari four cases of MMMT of the ovary were diagnosed. Three patients were in stage IIIC and one of them was a homologous MMMT; the fourth patient was affected by a heterologous stage IV MMMT. All women were treated with surgery and chemotherapy. Two patients are alive 14 and 12 months after diagnosis. The other two died after 37 months and one month, respectively. CONCLUSIONS: The malignant mixed mullerian tumor (MMMT) of the ovary is a particularly aggressive tumor, especially in advanced stages. The survival rate is very low in spite of surgery, chemotherapy and radiotherapy. The optimal treatment for this neoplasm is unknown because of its rarity. Our experience, when considering survival, seems to confirm the use of cisplatin and ifosfamide and to give new horizons to taxol.

Aggressive angiomyxoma of the pelvis. A case report.

Aggressive angiomyxoma is a locally mesenchymal, benign, and rare neoplasm. The vulva, perineum, and pelvis are the most common sites of involvement. The preoperative diagnosis is postulated by CT, sonography, MR image and angiography. The immunohistochemical study reveals the definitive diagnosis. The therapy is only surgical, and because of its tendency to recur locally, the excision has to be as complete as possible.

Recurrent peripartum cardiomyopathy.

Peripartum cardiomyopathy (PPCM) is an uncommon myocardial disease arising in the latter part of pregnancy or during the first five postpartum months, in the absence of any obvious cause and with no previously known heart disease. The risk of recurrence of PPCM is considered low when left ventricular size and function return to normal: We illustrate a case of peripartum cardiomyopathy recurred in subsequent pregnancy despite the rapid return to normal of heart size and function.

[Ultrasonic evaluation of endometrial changes induced by cyclic sequential hormone substitution therapy]. / Valutazione ecografica delle modificazioni dell'endometrio indotte dalla terapia ormonale sostitutiva sequenziale ciclica.

BACKGROUND: At present the hormonal replacement therapy on postmenopausal women with uterus needs the use of progestins additionally to estrogens, to eliminate the risk of endometrial hyperplasia and carcinoma connected with the use of estrogens alone. The check of the endometrium during these therapies can be made by transvaginal ultrasound that permits the evaluation of the thickness, structure, and contour of the endometrial rima. The aim of this study was to establish the changes of endometrial thickness during cyclic sequential hormonal replacement therapy on healthy postmenopausal women with transvaginal ultrasound. METHODS: The endometrial thickness with transvaginal ultrasound has been evaluated during the cyclic sequential hormonal replacement therapy on 20 healthy women in physiological menopause before the treatment, during the phase of treatment with estrogens alone and during the phase of treatment with the addition of the progestins. RESULTS: Significant differences during the estrogenic phase compared to before treatment have been underlined (5.7 mm vs 3.5 mm p = 0.002), but not during progestinic phase compared to estrogenic (6 mm vs 5.7 mm p = 0.712). CONCLUSIONS: Transvaginal ultrasound is a useful investigation to evaluate the modifications of the thickness and structure of the endometrium during hormonal replacement therapy and can help early diagnosis of endometrial diseases during these treatments.

[Incidence of climacteric syndrome]. / Incidenza della sindrome climaterica.

A retrospective study on 300 women who attended the menopausal out-patient clinic of Physiopathology of Human Reproduction at Bari University Hospital from 1994 to 1995, was carried out to evaluate the incidence of climacteric syndrome in premenopause together with its correlations with the social and cultural environment. The results don't show differences between spontaneous and surgical menopause, and minor differences with premenopause. No relationship has been discovered between the onset of the menarche and the onset of spontaneous menopause. The hot flushes are more frequent in women with low level of learning.

Fetal malformations in an epileptic pregnant woman treated with carbamazepine.

We report a case of an epileptic pregnant woman at 20 weeks of gestation treated with carbamazepine, whose fetus was affected by major malformations probably due to the drug teratogenicity. After a therapeutic abortion minor malformations were also found. After seven months the patient was pregnant again, at 14 weeks of gestation she aborted spontaneously a fetus with no malformations. During this pregnancy the patient took a reduced dosage of carbamazepine.

Karyotype abnormalities in a variant Chinese hamster cell line resistant to methyl methanesulphonate.

A variant cell population, isolated from V79-C13 Chinese hamster cells after two consecutive treatments with methyl methanesulphonate (MMS), was found to be highly resistant to killing by this alkylating agent. The resistant cell line was cytogenetically characterized both by the presence of a stable translocation involving metacentric chromosome 2 and acrocentric chromosome 6 and by a supernumerary chromosome originated by the duplication of a small telocentric chromosome. This cell population also showed a transient transformed phenotype, seen as formation of transformed foci containing cells with high chromosomes counts and multiple chromosomal aberrations. As MMS-resistance and karyotype changes are permanent and heritable traits, we suppose that they are related events.

The diagnosis of benign uterine pathology using transvaginal endohysterosonography.

Twenty-nine women believed to be affected with benign uterine pathologies underwent transvaginal hysterosonography insertion of a physiological solution into the cervical cavity by means of a catheter positioned in the cervical canal. This new technique, due to the acoustic window created by the fluid, made it possible to find: one cervical polyp, six endometrial polyps, one endometrial synechia, five submucosa myomas and one uterine malformation, the presence of which, with traditional ultrasonography can only be suspected. The subsequent hysteroscopic check-up confirmed the high diagnostic reliability of hysterosonography which is also easy to carry out, safe and costs little.

Possible involvement of the IL4 gene in Waldenström's macroglobulinemia.

We report the results of a molecular investigation of 11 patients affected by Waldenström's macroglobulinemia, a rare B-cell malignancy characterized by an excessive proliferation of immunoglobulin (Ig)M-secreting plasmacytoid cells. In particular, we studied the interleukin-4 (IL4) gene, which codes for a B-specific growth factor capable of stimulating the proliferation and differentiation of secreting plasma cells. By Southern hybridization, in three patients we found the presence of additional bands in comparison with the expected pattern; moreover, these bands showed a different degree of intensity.

Ultrasound diagnosis of heterotopic pregnancy with viable fetuses.

Combined pregnancy is an under-estimated and increasing clinical condition. Its recognition is usually made post-operatively. Ultrasound findings are not specific. Diagnosis is possible only when the cardiac activity of the extrauterine and intrauterine fetus can be detected. In the literature, 18 pre-operative sonographic demonstrations of such cases are reported. A correct echographic assessment can show a definite diagnosis or indicate a diagnostic laparoscopy minimizing maternal morbidity and mortality and improving the prognosis of the intrauterine fetus. In this report, ultrasound diagnostic aspects are discussed.

Sister chromatid exchange in Waldenström's macroglobulinemia.

Results on sister chromatid exchange (SCE) frequency and interchromosomal distribution in bone marrow and peripheral blood cultures from patients with Waldenström's macroglobulinemia are reported. PHA-stimulated bone marrow cultures showed increased SCE frequencies in all 12 patients examined. The increase was particularly high in two cases (17.07 and 16.77 SCE/cell, respectively) and, in one of them, a very high SCE level was found in PHA-stimulated peripheral blood culture (40.81 SCE/cell). In LPS-stimulated cultures, increased SCE levels were observed in some patients. Comparison between SCE frequency in bone marrow cell cultures with either mitogen showed a significant increase in PHA-stimulated cultures. Analysis of the interchromosomal SCE distribution revealed significant differences with respect to the control values; however, these differences were variable in the different patients. In pooled data of PHA-stimulated bone marrow cultures, there were differences between expected and observed SCEs in chromosomes 1 and 2 and in B, E, F, and G chromosome groups. Results of cell cycle modifications are also reported.

A new pericentric inv(8) in acute nonlymphocytic leukemia.

Chromosome studies were carried out on unstimulated bone marrow cells from a patient with a diagnosis of acute nonlymphocytic leukemia (FAB M6 ANLL). Cytogenetic analysis revealed a mosaic chromosome pattern 46,XX/46,XX,inv(8)(p21q24). This pericentric inversion has not been previously described in ANLL. Because, fragile sites, zinc finger gene loci, and the MYC protooncogene have been localized to band 8q24, a putative role for these sites and genes could be considered.

Chromosomal abnormalities in Waldenström's macroglobulinemia.

We report the results of cytogenetic studies of direct bone marrow (BM) preparations and of short-term BM and peripheral blood (PB) cultures from 17 patients with Waldenström's macroglobulinemia. We noted clonal chromosome changes in 10 patients. Abnormalities affected chromosomes X, Y, 2, 4, 5, 15, 16, 18, 19, 20, 21, and 22; in particular, chromosomes 2, 4, and 5 were involved in structural changes: a homogeneously staining region [hsr(2)], a der(4)t(4;?)(q32;?), and a 5q+. The other chromosomes were involved in numerical abnormalities, such as pseudodiploidy (a 46,X, -X, + 15 clone), loss of chromosome Y, and monosomy of chromosomes 16, 18, 19, 20, 21, and 22. Nonclonal chromosome rearrangements were also observed. The results are discussed in comparison with the few data reported in the literature, and the finding of an hsr in the long arm of chromosome 2 is emphasized; indeed, this is the first report of hsr in WM.

Use of intra-amniotic, intramuscular or intravaginal prostaglandins for mid-trimester termination of pregnancy.

PIP: 277 terminations of 2nd trimester pregnancies performed at the University Department of Obstetrics and Gynecology of Bari, Italy, during 1985-1990 with intraamniotic administration of hypertonic saline and 35 mg of PGF2alpha (Dinoprost) was the method used initially; more than 90% of cases (84 out of 91) were carried out before 1987. From the end f 1986, intramuscular Sulprostone was used at the dose of 500 mg every 3 hours, for a maximum of 3 injections in 87 cases. Vaginal administration of 1 mg Gemeprost suppository every 3 hours was introduced in 1988 and almost completely replaced the previous techniques (99 cases were reviewed). The mean age of patients was 26.3 years (range 15-47), and gestational age varied from 15 to 24 weeks. 169 (61%) were nulliparous, 97 (35%) had 1 to 3 children, and 11 patients had more than three. 139 (50.2%) were married. Regarding previous terminations of pregnancy, 235 patients (84.8%) had had none, 23 had had one, and 19 more than one. 128 (46.2%) patients came from low socioeconomic class, 105 (37.9%) were middle class, and 44 were high class. The statistical analysis ANOVA showed abortion induction interval of 24.02, 24,28, and 14.24 hours, respectively for the intraamniotic, the sulprostone, and the Gemeprost methods. The days of hospitalization were 9.57, 8.50, and 11.18, respectively. Psychological evaluation indicated that Gemeprost was preferred by the patients, while the intraamniotic method was perceived negatively. Side-effects were significantly milder, thus Gemeprost is considered the first choice for induction of mid-trimester abortion after evaluation of possible contraindications to prostaglandins. In those cases where severe stenosis of the cervical canal arises, laminaria pretreatment followed by either Gemeprost or Sulprostone is highly effective.^ieng