Probing lysine mono-methylation in histone H3 tail peptides with an abiotic receptor coupled to a non-plasmonic resonator.

Binder and effector molecules that allow studying and manipulating epigenetic processes are of biological relevance and pose severe technical challenges. We report the first example of a synthetic receptor able to recognize mono-methylated lysines in a histone H3 tail peptide, which has relevant functions in epigenetic regulation. Recognition is robust and specific regardless of the position and the number of mono-methylated lysines along the polypeptide chain. The peptide is first captured in solution by a tetraphosphonate cavitand (Tiiii) that selectively binds its Lys-NMe+ moieties. Separation from solution and detection of the peptide-Tiiii complexes is then enabled in one single step by an all dielectric SiO2-TiO2 core-shell resonator (T-rex), which captures the complex and operates fully reproducible signal transduction by non-plasmonic surface enhanced Raman scattering (SERS) without degrading the complex. The realized abiotic probe is able to distinguish multiple mono-methylated peptides from the single mono-methylated ones.

Unraveling the role of mechanical stimulation on smooth muscle cells: A comparative study between 2D and 3D models.

A thorough understanding of cell response to combined culture configuration and mechanical cues is of paramount importance in vascular tissue engineering applications. Herein, we investigated and compared the response of vascular smooth muscle cells (vSMCs) cultured in different culture environments (2D cell monolayers and 3D cellularized collagen-based gels) in combination with mechanical stimulation (7% uniaxial cyclic strain, 1 Hz) for 2 and 5 days. When cyclic strain was applied, two different responses, in terms of cell orientation and expression of contractile-phenotype proteins, were observed in 2D and 3D models. Specifically, in 2D configuration, cyclic strain caused ∼50% of cell population to align nearly perpendicular (80-90 degrees) to the strain direction, while not influencing the contractile-phenotype protein expression, as compared to the 2D static controls. Conversely, the application of uniaxial strain to 3D constructs induced a ∼60% cell alignment almost parallel (0-10 degrees) to the strain direction. Moreover, 3D mechanical stimulation applied for 5 days induced a twofold increase of SM α-actin level and a 14-fold increase of calponin expression as compared to 3D static controls. Altogether these findings provide a new insight into the potential to drive cell behavior by modulating the extracellular matrix and the biomechanical environment. Biotechnol. Bioeng. 2016;113: 2254-2263. © 2016 Wiley Periodicals, Inc.

Data in support of Gallium (Ga(3+)) antibacterial activities to counteract E. coli and S. epidermidis biofilm formation onto pro-osteointegrative titanium surfaces.

This paper contains original data supporting the antibacterial activities of Gallium (Ga(3+))-doped pro-osteointegrative titanium alloys, obtained via Anodic Spark Deposition (ASD), as described in "The effect of silver or gallium doped titanium against the multidrug resistant Acinetobacter baumannii" (Cochis et al. 2016) [1]. In this article we included an indirect cytocompatibility evaluation towards Saos2 human osteoblasts and extended the microbial evaluation of the Ga(3+) enriched titanium surfaces against the biofilm former Escherichia coli and Staphylococcus epidermidis strains. Cell viability was assayed by the Alamar Blue test, while bacterial viability was evaluated by the metabolic colorimetric 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay. Finally biofilm morphology was analyzed by Scanning Electron Microscopy (SEM). Data regarding Ga(3+) activity were compared to Silver.

Chondrocyte response to high regimens of cyclic hydrostatic pressure in 3-dimensional engineered constructs.

PURPOSE: Despite widespread use of 3-dimensional (3D) micro-porous scaffolds to promote their potential application in cartilage tissue engineering, only a few studies have examined the response to hydrostatic pressure of engineered constructs. A high cyclic pressurization, currently believed to be the predominant mechanical signal perceived by cells in articular cartilage, was used here to stimulate bovine articular chondrocytes cultured in a synthetic 3D porous scaffold (DegraPol). METHODS: Construct cultivation lasted 3 days with applied pressurization cycles of amplitude 10 MPa, frequency 0.33 Hz, and stimulation sessions of 4 hours/day. RESULTS: At 3 days of culture, with respect to pre-culture conditions, the viability of the pressurized constructs did not vary, whereas it underwent a 16% drop in the unpressurized controls. Synthesis of alfa-actin was 34% lower in all cultured constructs. Synthesis of collagen II/collagen I did not vary in pressurized constructs, was 76% lower in unpressurized controls, and was around 230% higher in pressurized constructs with respect to unpressurized controls. Chondrocytes showed a phenotypic spherical morphology at time zero and at 3 days of pressurized culture. CONCLUSIONS: Although the passage from 2D expansion to 3D geometry was effective to guide cell differentiation, only mechanical conditioning enabled the maintenance and further cell differentiation toward a mature chondrocytic phenotype.

Gene therapy: The state of the art and future directions.

Gene therapy, because of its aim to eradicate the causes rather than the symptoms of diseases, is believed by many to be the therapy of the future. The problems of developing clinically viable gene therapeutic approaches and designing safe and efficient gene delivery reagents are inseparable: shortcomings in one are going to affect adversely the success of the other. It is generally accepted that the major impediment to the successful application of gene therapy for the treatment of a range of dis-eases is not a paucity of therapeutic genes, but the lack of efficient non-toxic gene delivery systems. Transfection vectors com-monly used in gene therapy are mainly of two types: viral and non-viral. Non-viral gene delivery is currently the subject of increasing attention because of its relative safety and its simplicity of use; however, its use is still far from being ideal due to its comparatively low efficiency. The purpose of this review is to give an overview of the current knowledge concerning the assembly of lipoplexes and the trafficking of lipoplexes into cells, as well as to underscore the advantages and disadvantages of lipidic gene carriers among non-viral gene delivery systems.

Tumor necrosis factor alpha and glutathione interplay in chronic heart failure.

The pro-inflammatory cytokine, tumor necrosis factor alpha (TNF alpha), in concert with neurohormones, contributes to chronic heart failure (CHF) progression. This implies that TNF a antagonism may constitute an important target for CHF therapy. However, clinical trials in CHF patients using compounds that trap TNF alpha, comprising infliximab, an antibody directed to TNF alpha, and etanercept, a soluble recombinant receptor of TNF alpha, gave disappointing results bringing back to light the dual, short-term beneficial and long-term harmful effect of TNF alpha. This review focuses on the dual, concentration- and time-related effects of TNF alpha, the yin and yang action of TNF alpha in cardiac ischemia/reperfusion and contraction. Importantly, the harmful effects of TNF a are related to glutathione deficiency, a common hallmark to several other chronic inflammatory diseases. Recently, in rat models of CHF, oral administration of the glutathione precursor, N-acetylcysteine (NAC), was shown to hinder pathways of TNF alpha harmful signalling and to rescue cardiac structure and function. These results suggest that glutathione deficiency in association with TNF alpha activation may play a role in the pathophysiology of CHF and that NAC may represent a potential therapy in CHF.

Chrysanthones, a new source of fungal metabolites with potential antitumor and antiangiogenesis properties.

Chrysanthones are secondary metabolites, isolated from Ascochyta chrysanthemi, with a methyl-benzoisoquinoline or methyl-benzoisochromene system. Three compounds were tested for their cytotoxicity properties on endothelial cells (EC) and two different tumor cell lines and for their ability to inhibit EC migration. Structure-function relationship considerations suggested that the methylisoquinolinic moiety is important for the cytotoxic activity, whereas the methylisochromene moiety confers an endothelial selectivity to the structures. In general, compared to the activity of known antiproliferative and antiangiogenic compounds, chrysanthones showed weaker activities. However, they present a basis for the synthesis of new derivatives.

In-vitro and in-vivo antibacterial activity of BI 397, a new semi-synthetic glycopeptide antibiotic.

BI 397 (formerly A-A-1) is a semisynthetic derivative of the teicoplanin-like glycopeptide A40926. It was more active in vitro against staphylococci (including some teicoplanin-resistant strains) than teicoplanin and vancomycin. Against streptococci (including penicillin-resistant strains) BI 397 has activity comparable with that of teicoplanin and better than vancomycin. BI 397, when administered to rats by the i.v. route, gives high and long lasting blood levels. It shows excellent activity in models of acute septicaemia in immunocompetent and neutropenic mice. In a rat staphylococcal endocarditis model it is as effective as teicoplanin and vancomycin at reducing bacterial loads in the heart, but at lower dosages and with a reduced number of daily treatments compared with the two glycopeptide controls. BI 397 is highly efficacious in clearing penicillin-susceptible and -resistant pneumococci from lungs of immunocompetent and neutropenic rats. The data from these studies show that BI 397 combines an excellent in-vitro antibacterial activity with favourable pharmacokinetic behaviour resulting in potent in-vivo activity.

Does aggregation substance of Enterococcus faecalis contribute to development of endocarditis?

Aggregation substance (AS) of Enterococcus faecalis which is encoded by so-called sex pheromone plasmids enables the bacteria to bind to in vitro-cultured pig kidney tubular cells. It is reported that the presence of AS is not of pivotal importance for the ability of E. faecalis to cause infective endocarditis (EN). The lines of evidence for this are twofold: 1) sex pheromone plasmids and, therefore, the gene for AS were not present more often in epidemiologically unrelated strains of E. faecalis isolated from human cases of EN than in isolates from well-water (26 vs. 18%); 2) the presence of the adhesin did not correlate with the establishment of EN in an animal (rat) model. The data are discussed with respect to the specificity of interaction of AS with eukaryotic cells and the results of other studies.

Double uterus, blind hemivagina, and ipsilateral renal agenesis: 36 cases and long-term follow-up.

OBJECTIVE: To review the experience of the Milan University First Department of Obstetrics and Gynecology in patients with double, didelphic, of bicornuate uterus, blind hemivagina, and ipsilateral renal agenesis, and to consider the frequently unsatisfactory surgical approach. METHODS: Thirty-six women with double, didelphic, or bicornuate uterus, blind hemivagina, and ipsilateral renal agenesis were identified from clinical records for the period 1962 to 1992. We evaluated demographic data, disease, symptoms, correctness of therapeutic approach, and definitive treatment. RESULTS: Seventeen patients previously had undergone incomplete surgery in other hospitals and 19 were treated by us for the first time. Total hysterectomy was performed on two of the 36 women and hemihysterectomy and hemicolpectomy were performed on four. In the other 30, the vaginal septum was excised and marsupialization was done. The pregnancy rate in the 15 women wanting children was 87% and the live birth rate was 77%. Serial biopsy specimens were obtained from the lateral fornix after the excision of the septum in 13 of the non-hysterectomized patients over 1-9 years and revealed progressively more extensive areas of squamous metaplasia of müllerian epithelium. In some isolated cases, papillary hyperplasia, mild dysplasia, and vaginal adenosis were found. At the end of follow-up, 16 patients still did not want children. Follow-up was possible in 34 cases. CONCLUSION: Early accurate diagnosis after menarche followed by excision and marsupialization of the blind hemivagina offers complete relief of symptoms and preserves reproductive potential. Partial morphologic changes are evident but metabolic modifications comparable to those of the adjacent normal vagina have not yet been documented.

Experimental lobar pneumonia due to penicillin-susceptible and penicillin-resistant Streptococcus pneumoniae in immunocompetent and neutropenic rats: efficacy of penicillin and teicoplanin treatment.

Lobar pneumonia models were established in rats by intratracheal inoculation of either penicillin-susceptible (immunocompetent model) or penicillin-resistant (immunocompetent and neutropenic models) Streptococcus pneumoniae. Untreated animals maintained a relatively high bacterial load in the lungs but only occasionally developed bacteraemia or pleurisy. The infection was rarely fatal in immunocompetent rats, but immunocompromised rats frequently died. Treatment i.m. with 10,000 IU/kg of procaine penicillin G (12 h after infection then bid for 3 days) or with a single i.v. dose of 5 or 10 mg/kg of teicoplanin significantly reduced lung bacterial loads of rats infected with the penicillin-susceptible strain. Against the penicillin-resistant strain, teicoplanin displayed a significant activity regardless of the immununological status of the animals. Penicillin G significantly reduced lung bacterial load of the penicillin-resistant strain only in immunocompetent rats and at a higher dose than needed in treatment of the penicillin-susceptible infection. The experimental models described here could be suitable for studying the efficacy of antibacterial agents against pulmonary infections caused by penicillin-susceptible and penicillin-resistant S. pneumoniae strains.

Posthysterectomy fallopian tube herniation. A report of two cases.

BACKGROUND: Tubal herniation as a complication of hysterectomy is a rare phenomenon, markedly more frequent after vaginal hysterectomy. With the increasing use of the vaginal route, the ratio between tubal herniation after vaginal versus abdominal hysterectomy may exceed 3:1. CASE: We report two cases of tubal herniation into the vagina, one after vaginal hysterectomy and the other after total abdominal hysterectomy, in two patients, aged 36 and 37 years. CONCLUSION: A tubal prolapse in the vagina may be considered a hernia and occurs only if a communication exists between the peritoneal cavity and vaginal canal. It can be an early or late prolapse. Symptoms consist almost exclusively of persistent blood loss and/or leukorrhea, dyspareunia and chronic pelvic pain. Whether the abdominal or vaginal approach should be used in surgical correction of prolapsed tubes must be decided in each case according to the patient's individual characteristics. Both histologic pictures described merit careful attention, distinguishing between the terminal tubal segment and the more cranial tract (above the vaginal strangulation).

Antimicrobial activity of MDL 63,246, a new semisynthetic glycopeptide antibiotic.

MDL 63,246 is a semisynthetic derivative of the naturally occurring glycopeptide antibiotic MDL 62,476 (A40926). It was more active in vitro against Staphylococcus aureus and coagulase-negative staphylococci than MDL 62,476, teicoplanin, and vancomycin and was more active than mideplanin (MDL 62,873) against some isolates. MDL 63,246 had excellent activity against streptococci and teicoplanin-susceptible enterococci, and it also had in vitro activity against some VanA enterococcal isolates. It was more active than teicoplanin and vancomycin against acute staphylococcal, streptococcal, and enterococcal septicemia in immunocompetent and neutropenic mice. It was highly efficacious in reducing the bacterial load in the hearts of rats in staphylococcal endocarditis experiments and the bacterial load of Staphylococcus epidermis in a high infection model in neutropenic mice. The excellent in vivo activity of MDL 63,246 appears to correlate both with its in vitro antibacterial activity and with its long half-life in rodents.

Fertility after conservative surgery for adenomyomas.

Reproductive performance was evaluated after conservative surgery for uterine adenomyoma in a prospective, observational study. Twenty-eight women with histologically proven adenomyomas had conservative surgery in the period 1985-1990. Eighteen patients wanting children were regularly followed for a mean +/- SD period of 53.2 +/- 23.5 months. Crude and cumulative pregnancy rates were calculated after the operation. Thirteen (72.2%) women conceived, for a total of 18 pregnancies: nine (50%) ended in term deliveries, seven (38.8%) in spontaneous abortions, one (5.6%) in ectopic pregnancy and one (5.6%) in a pre-term delivery with neonatal death. The cumulative pregnancy rate at 36 months of follow-up was 74.7%. This analysis of a small series indicates that conservative surgery for adenomyomas is associated with a good reproductive prognosis.

Successive transperitoneal migration of ova in a woman with extensive pelvic adhesions.

Our case report describes three conceptions after transperitoneal migration of the ovum in a woman with only one ovary, the contralateral oviduct, and extensive postoperative pelvic adhesions obliterating the Douglas cul-de-sac. This suggests that anatomic integrity of the pelvis is not always essential for ovum pick-up.

N63-carboxamides of N15-alkyl and N15,N15-dialkyl derivatives of teicoplanin and deglucoteicoplanin.

The synthesis and biological properties of a series of N63-carboxamides of 15-N-alkylated derivatives of teicoplanin A2 (CTA) and its aglycone (TD) are described. Among the compounds, those carrying hydrophilic groups or ionizable amino functions on the N15-alkyl chain are more soluble in water than parent N15-methylated or unmodified amides. Selected compounds were more active in vitro than CTA or TD, and a few of them were also slightly more efficacious in vivo than the parent antibiotics in streptococcal septicemia in the mouse. Their degree of activity varied with the structure and length of the N15-alkyl chains.

Synthesis and antibacterial activity of a series of basic amides of teicoplanin and deglucoteicoplanin with polyamines.

Basic carboxamides of teicoplanin A2 (CTA) and its aglycon (TD) are prepared by condensation of the 63-carboxyl function of these antibiotics with linear or branched polyamines. The antimicrobial activities of some of the resulting compounds were better than those of the unmodified antibiotics. The presence of more than one basic group in the amidic chain enhanced the in vitro activity of some TD-amides against Gram-negative bacteria; two of these derivatives were also effective in vivo against Escherichia coli septicemia in the mouse. Among the CTA derivatives, the amide with spermine showed some unexpected in vitro activity against Gram-negatives. Both CTA- and TD-amides with polyamines are very soluble in water over a wide range of pH and are very hydrophilic.

Presacral neurectomy for the treatment of pelvic pain associated with endometriosis: a controlled study.

OBJECTIVE: Our objective was to evaluate the efficacy of presacral neurectomy combined with conservative surgery for the treatment of pelvic pain associated with endometriosis. STUDY DESIGN: In a randomized, controlled study performed in a tertiary institution 71 patients with moderate or severe endometriosis and midline dysmenorrhea were randomly assigned to conservative surgery alone (n = 36) or conservative surgery and presacral neurectomy (n = 35). Main outcome measures were relief of dysmenorrhea, pelvic pain, and deep dyspareunia after surgery according to a multidimensional and an analog pain scale. RESULTS: Presacral neurectomy markedly reduced the midline component of menstrual pain, but no statistically significant differences were observed between the two groups in the frequency and severity of dysmenorrhea, pelvic pain, and dyspareunia in the long-term follow-up. After presacral neurectomy, constipation developed or worsened in 13 patients and urinary urgency occurred in three and a painless first stage of labor in two. CONCLUSION: Presacral neurectomy should be combined with conservative surgery for endometriosis only in selected cases.

Buserelin acetate versus expectant management in the treatment of infertility associated with minimal or mild endometriosis: a randomized clinical trial.

OBJECTIVE: We performed a randomized clinical trial to evaluate the efficacy of intranasal 400 micrograms buserelin three times daily for 6 months versus expectant management in the treatment of infertile women with pelvic endometriosis stage I or II of the revised American Fertility Society classification. STUDY DESIGN: Seventy-one consecutive patients (mean age 32 years) were studied at the First Department of Obstetrics and Gynecology, University of Milan, and the Department of Obstetrics and Gynecology, Ospedali Riuniti, Bergamo, between February 1988 and June 1989. Thirty-five women were randomly allocated to buserelin treatment and 36 to expectant management. The baseline distribution of subjects for age, disease stage, and reproductive history was similar in the two groups. All patients were followed regularly; median follow-up was 17 months in the buserelin group and 18 months in the women given expectant management. If pregnancy did not occur within 12 months of randomization, cycles were monitored by ultrasonography and hormone measurements, and when abnormalities were detected clomiphene citrate and human chorionic gonadotropin were administered. RESULTS: A total of 17 pregnancies were observed both in the buserelin-treated patients and in the expectant management group. The 1- and 2-year actuarial overall pregnancy rates were similar in the two groups, 30% and 61% in the former and 37% and 61% in the latter group, respectively. Spontaneous abortion occurred in five of the 17 pregnancies in the women treated with buserelin and in one of the 17 in those managed expectantly; this difference was, however, not statistically significant (chi 1(2) adjusted for disease stage and use of clomiphene citrate and human chorionic gonadotropin treatment = 3.01, p = 0.08). No fetal death or stillbirth was observed. CONCLUSIONS: Our findings suggest that treatment with gonadotropin-releasing hormone agonists is unlikely to have a marked influence on the reproductive outcome of infertile women with minimal or mild endometriosis.