Characterization of polymorphic ampicillin forms.

In this work polymorphs of α-aminobenzylpenicillin (ampicillin), a ß-lactamic antibiotic, were prepared and investigated by several experimental and theoretical methods. Amorphous monohydrate and three crystalline forms, the trihydrate, the crystal form I and the crystal form II, were investigated by FT-IR and micro-Raman. Also data obtained by differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), X-ray powder diffraction (XRPD) and hot-stage Raman spectroscopy are reported. Finally, quantum mechanical calculations were performed by density functional theory (DFT) to assist the assignment of spectroscopic experimental bands. For the first time, the ampicillin molecule in its zwitterionic form was studied at the B3LYP/aug-cc-pVDZ level and the corresponding theoretical vibrational spectra were computed. In fact, ampicillin in the crystal is in zwitterionic form and concentrations of this same form are quite relevant in solutions at physiological pH. Experimental and theoretical results allowed identification of specific features for polymorph characterization. Bands typical of the different polymorphs are identified both in IR and Raman spectra: in particular in the NH stretching region (IR), in the amide I+δNH region (both techniques), in the 1520-1490cm(-1) region (IR), in the 1320-1300cm(-1) and 1280-1220cm(-1) (IR), in the 1200-1170cm(-1) (Raman), in the amide V region (IR), and, finally, in the 715-640cm(-1) and 220-200cm(-1) (Raman). Interconversion among different polymorphs was investigated by hot-stage Raman spectroscopy and thermal analysis, clarifying the complex pattern of transformations undergone as a function of temperature and heating rate. In particular, DSC scans show how the trihydrate crystals transform into anhydrous forms on heating. Finally, stability tests demonstrated, after a two years period, that no transformation or degradation of the polymorphs occurred.

IR, Raman and SERS spectra of propantheline bromide.

The two known propantheline bromide polymorphs (form I and form II) were studied and characterized by a multianalytical approach. In the present work, the identification of propantheline bromide polymorphic forms through vibrational IR spectroscopies are presented and for the first time Raman microscopy and hot stage Raman microscopy (HSRM) studies are reported. Finally, quantum mechanical calculations were performed. For assisting the assignment of the experimental picks, the two IR spectra of the most and least stable representatives of a set of 56 conformers are calculated and studied. DSC thermograms data, are also reported. The surface enhanced Raman scattering (SERS) spectrum was also recorded in a silver colloid; it could be inferred that propantheline bromide is adsorbed on silver colloid through the oxygen atom with the molecular plane perpendicular to the metal surface.

Use of Raman spectroscopy for the identification of radical-mediated damages in human serum albumin.

Damages induced by free radicals on human serum albumin (HSA), the most prominent protein in plasma, were investigated by Raman spectroscopy. HSA underwent oxidative and reductive radical stress. Gamma-irradiation was used to simulate the endogenous formation of reactive radical species such as hydrogen atoms ((•)H), solvated electrons (e(aq)(-)) and hydroxyl radicals ((•)OH). Raman spectroscopy was shown to be a useful tool in identifying conformational changes of the protein structure and specific damages occurring at sensitive amino acid sites. In particular, the analysis of the S-S stretching region suggested the radical species caused modifications in the 17 disulphide bridges of HSA. The concomitant action of e(aq)(-) and (•)H atoms caused the formation of cyclic disulphide bridges, showing how cystine pairs act as efficient interceptors of reducing species, by direct scavenging and electron transfer reactions within the protein. This conclusion was further confirmed by the modifications visible in the Raman bands due to Phe and Tyr residues. As regards to protein folding, both oxidative and reductive radical stresses were able to cause a loss in α-helix content, although the latter remains the most abundant secondary structure component. ß-turns motifs significantly increased as a consequence of the synergic action of e(aq)(-) and (•)H atoms, whereas a larger increase in the ß-sheet content was found following the exposure to (•)OH and/or (•)H attack.

Apatite-forming ability (bioactivity) of ProRoot MTA.

AIM: Apatite-forming ability, considered as an index of bioactivity (bond-to-bone ability), was tested on ProRoot MTA cement after immersion in phosphate-containing solution (DPBS). METHODOLOGY: Disk samples were prepared and immersed in DPBS for 10 min, 5 h, 1 and 7 days. The cement surface was studied by attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy, by micro-Raman spectroscopy and by environmental scanning electron microscope with energy dispersive X-ray (ESEM-EDX) analyses. The pH of the storage solution was also investigated. RESULTS: Spectroscopic analyses revealed calcium phosphate bands after 5-h immersion in DPBS. After 1 day, an even coating composed of apatite spherulites (0.1-0.8 micron diameter) was observed by ESEM/EDX. After 7 days, its thickness had increased. Apatite nucleation had already occurred after 5-h immersion. At this time, the presence of portlandite (i.e. Ca(OH)(2) , calcium hydroxide) on the cement surface was also observed; at longer times, this component was released into the medium, which underwent a remarkable pH increase. CONCLUSIONS: The study confirms the ability of ProRoot MTA to form a superficial layer of apatite within hours. The excellent bioactivity of ProRoot MTA might provide a significant clinical advantage over the traditional cements used for root-end or root-perforation repair.

Comparison between the in vitro surface transformations of AP40 and RKKP bioactive glasses.

Two bioactive silica-phosphate glasses, AP40 and RKKP, were compared in their behaviour in simulated biological environment. Their chemical composition is practically identical, except that RKKP contains small amounts of amphoteric network-former oxides Ta2O5 and La2O3 (composition in wt% for AP40: beta-Ca3(PO4)2 24.50, SiO2 44.30, CaO 18.60, Na2O 4.60, K2O 0.19, MgO 2.82, CaF2 4.99; RKKP: beta-Ca3(PO4)2 24.23, SiO2 43.82, CaO 18.40, Na2O 4.55, K2O 0.19, MgO 2.79, CaF2 4.94, Ta2O5 0.99, La2O3 0.09). Previous investigations showed a better performance in osteopenic bone for RKKP. To gain more insight into these differences in biological behaviour, the in vitro bioactivity of the glasses was studied by treatment with a continuously replenished Hanks' Balanced Salt Solution (HBSS). The glasses were examined before and after HBSS treatment for 20 and 40 days by X-ray Diffraction (XRD), Scanning Electron Microscopy (SEM), X-ray Energy Dispersion (EDX), Raman and IR vibrational spectroscopies. Some slight but notable differences between the two glasses were observed after HBSS treatment. IR and EDX analyses showed that deposits formed on both glasses were composed of a calcium deficient carbonate-apatite; however, the layer formed on RKKP glass was found to be slightly more calcium deficient and thinner. EDX analysis evidenced the presence of a small percentage of F- ions only in the layers formed on the RKKP samples. The differences disclosed, although slight, can contribute to the understanding of the different biological behaviour previously observed.

Spectroscopic behavior of copper complexes of nonsteroidal anti-inflammatory drugs.

The proposed curative properties of Cu-based nonsteroidal anti-inflammatory drugs (NSAIDs) have led to the development of numerous Cu(II) complexes of NSAIDs with enhanced anti-inflammatory activity. Crystalline complexes, Cu(II)-NSAID (ibuprofen, naproxen, tolmetin, and diclofenac), with a carboxylic function have been studied by means of infrared and Raman spectroscopy. All NSAIDs bind the metal through the carboxylate group. On the basis of the comparison between the wavenumber of the COO(-) group vibrations and Delta nu (nu(asimm)COO(-) - nu(simm)COO(-)) between Na salts and Cu(II) complexes, conclusions on the probable structure of the complexes have been drawn. The spectroscopic data support the formation of dimeric [Cu(2)L(4)(H(2)O)(2)] complexes in which the COO(-) group behaves as a bridging bidentate ligand. The low wavenumber region of the Raman spectrum provided information on Cu-O and Cu-Cu bonds in the complexes. Thermogravimetric results gave further support to the vibrational data.

[172 consecutive total gastrectomies for gastric cancer with Roux-en-Y loop reconstruction and without fistulization of the esophago-jejunal anastomosis]. / 172 gastrectomie totali consecutive per cancro gastrico con ricostruzione su ansa alla roux e senza fistolizzazione dell'anastomosi esofagodigiunale.

In the treatment of gastric cancer total gastrectomy (TG) is one of the most important operation and the esophagojejunal anastomotic leakage is the most important early complication. In our series of 172 consecutive TGs (all controlled with Gastrografin-Rx in fifth postoperative day) we did not have any anastomotic fistula, independently from age, stage, type of limphadenectomy and visceral demolition. We believe that the correct technical performance is the most important factor in the esophagojejunal anastomosis.

[Reliability of extemporaneous histological sentinel lymph node examination during breast surgery for neoplasia]. / Affidabilità dell'esame istologico estemporaneo del linfonodo sentinella in corso di chirurgia mammaria per neoplasia.

Lymphatic mapping and sentinel lymph node (SLN) biopsy for breast cancer is rapidly becoming the standard of care. This is mainly due to the accuracy of the procedure, with a significant decrease in morbidity compared with the standard level III node dissection. We present our experience with SLN biopsy and a small series in which we performed an immediate histologic evaluation of the SLN: in case of positive SLN, a complete lymph node dissection was carried out in the same operative time, thus reducing the need of a second operation. In our experience, we had a 100% accordance between immediate and definitive results: we had neither false positive, which could lead to overtreat the patients with an unnecessary lymph node dissection, nor false negative.

1H-NMR and 13C-NMR lipid profiles of human renal tissues.

Lipids from human renal tissues are studied by means of (1)H- and (13)C-NMR spectroscopy. The total lipid fractions obtained from healthy kidneys, malignant renal cell carcinomas, and benign oncocytomas are characterized and analyzed to elucidate the main differences between the functional and neoplastic tissues. In all cases the lipid components are well identified. The healthy kidney is characterized by high amounts of triglycerides and the presence of cholesterol in its free form. On the contrary, renal cell carcinomas contain high amounts of cholesterol that are almost completely esterified as oleate, suggesting an intracellular localization of the cholesteryl esters synthesis. Cholesteryl esters are considered markers of renal cell carcinomas, thus supporting recent theories that these compounds play a leading role in cell proliferation. Oncocytomas are particularly rich in phosphatidylcholine and, analogous to the healthy kidney, are completely lacking in cholesteryl esters. Healthy kidneys and oncocytomas appear to have other similarities if compared with renal cell carcinomas: a very high fatty acyl/cholesterol ratio, the presence of dolichols, and a higher grade of unsaturation. The (13)C data suggest a new method for the direct evaluation of the saturated/unsaturated fatty acyl ratio.

Detachment of titanium and fluorohydroxyapatite particles in unloaded endosseous implants.

The shape, surface composition and morphology of orthopaedic and endosseous dental titanium implants are key factors to achieve post-surgical and long-term mechanical stability and enhance implant osteointegration. In this study a comparison was made between 12 titanium screws, plasma-spray-coated with titanium powders (TPS), and 12 screws with an additional coating of fluorohydroxyapatite (FHA-Ti). Screws were implanted in the femoral and tibial diaphyses of two mongrel sheep and removed with peri-implant tissues 12 weeks after surgery. The vibrational spectroscopic, ultrastructural and morphological analyses showed good osteointegration for both types of implants in host cortical bone. The portion of the FHA-Ti implants in contact with the medullary canal showed a wider area of newly formed peri-implant bone than that of the TPS implants. Morphological and EDAX analyses demonstrated the presence of small titanium debris in the bone medullary spaces near the TPS surface, presumably due to the friction between the host bone and the implant during insertion. Few traces of titanium were detected around FHA-Ti implants, even if smaller FHA debris were present. The present findings suggest that the FHA coating may act as a barrier against the detachment of titanium debris stored in the medullary spaces near the implant surface.

Effects of the sterilisation method on the wear of UHMWPE acetabular cups tested in a hip joint simulator.

Ultra-high molecular-weight-polyethylene is the most commonly used bearing material in total joint replacement. Wear of polyethylene is a Serious Clinical problem that limits the longevity of orthopaedic implants. Information on degradative changes in the material properties and on the methods used for the sterilisation of polyethylene may help in the selection process of orthopaedic implants with the best wear resistance. This study was performed to investigate the effects of the sterilisation method (gamma irradiation and ethylene oxide treatment) on the wear and on the changes in physical properties of polyethylene acetabular cups. At this purpose, gamma-sterilised and ethylene oxide (EtO)-sterilised acetabular cups were tested against CoCr femoral heads in a hip joint simulator run for 5 million cycles in bovine calf serum. The crystallinity of the cups was evaluated by micro-Raman spectroscopy as a function of the inner surface position. The partial least square calibration was used to correlate the Raman spectra with the crystallinity of the polymer measured by differential scanning calorimetry. The analysis performed on soak control acetabular cups demonstrated that the gamma-sterilised cups are significantly more crystalline than the EtO-sterilised ones. The mean crystallinity values obtained for the gamma-sterilised and EtO-sterilised soak control cups were 65.0% and 63.4%, respectively. Weight loss measurements revealed that the gamma-sterilised acetabular cups exhibited a lower wear rate than that by EtO-sterilised. Thc Raman results obtained on gamma-sterilised and EtO-sterilised acetabular cups showed that the changes in surface crystallinity were mainly caused by irradiation rather than by the mechanical friction during the in vitro tests.

Molecular characterization of human healthy and neoplastic cerebral and renal tissues by in vitro (1)H NMR spectroscopy (review).

The clinical impact of (1)H NMR spectroscopy in the study of human organs, brain and kidney in particular, is well demonstrated. The in vitro (1)H NMR technique is a powerful tool for monitoring changes in intracellular metabolites of human normal and neoplastic cerebral and renal tissues. Healthy and tumoral tissues of different histologic types have been fully characterized from a biochemical standpoint. Molecular characterization is performed on both the aqueous and lipid extracts of surgically removed tissue biopsies yielding a full picture of tissue biochemistry. These analyses have disclosed markers of healthy brain and kidney and of their respective neoplastic lesions. Moreover, some biochemical features can differentiate neoplasms within the same histological type. In particular, lipidic components, like cholesteryl esters (namely oleate), detected in highest grade tumors, warrant further investigation. A better understanding of the biochemistry of diseased human tissues could open the way to new diagnostic and treatment strategies.

Congenital defect in the abdominal wall in an adult: a case report.

We describe a rare case of a woman aged 38 who presented with an untreated omphalocele in association with a duplication of the vena cava and malposition of the right kidney. An aesthetic and functional reconstruction of the abdomen was achieved with local flaps and the creation of a neoumbilicus.

Study of the vibrational spectrum of cuprorivaite.

In this research, after tracing a history of the pigment, the preparation and analysis of samples of Egyptian blue have been reported. The main reactions taking place during its preparation are indicated. X-ray diffractions show that, starting from mixtures of precursors with stoichiometric composition, the product is practically only cuprorivaite. The kind of flux and cristallinity of silica does not seem determining factors for the pigment formation. Infrared and Raman spectra and an assignment of the bands are reported. Impurities ascertained are discussed on the basis on the data available.

Characterization of lipids from human brain tissues by multinuclear magnetic resonance spectroscopy.

Multinuclear ((1)H, (13)C, and (31)P) magnetic resonance spectroscopy are applied to the biochemical characterization of the total lipid fraction of healthy and neoplastic human brain tissues. Lipid extracts from normal brains, glioblastomas, anaplastic oligodendrogliomas, oligodendrogliomas, and meningiomas are examined. Moreover, the unknown liquid content of a cyst adjacent to a meningioma is analyzed. Two biopsies from glioblastomas are directly studied by (1)H-NMR without any treatment (ex vivo NMR). The (1)H- and (13)C-NMR analysis allows full characterization of the lipid component of the cerebral tissues. In particular, the presence of cholesteryl esters and triglycerides in the extracts of high grade tumors is correlated to the vascular proliferation degree, which is different from normal brain tissue and low grade neoplasms. The (31)P spectra show that phosphatidylcholine is the prominent phospholipid and its relative amount, which is higher in gliomas, is correlated to the low grade of differentiation of tumor cells and an altered membrane turnover. The ex vivo (1)H-NMR data on the glioblastoma samples show the presence of mobile lipids that are correlated to cell necrotic phenomena. Our data allow a direct correlation between biochemical results obtained by NMR and the histopathological factors (vascular and cell proliferations, differentiation, and necrosis) that are prominent in determining brain tumor grading.

Vibrational spectroscopic characterization of new calcium phosphate bioactive coatings.

In this work calcium phosphate (CaP) compounds with different PO(3-)(4)/HPO(2-)(4) R molar ratios in the 0.65-149 range were synthesized. In fact, all these CaPs contain different amounts of HPO(2-)(4) and PO(3-)(4) ions as well as the amorphous precursors (tricalcium phosphate and octacalcium phosphate) of hydroxyapatite deposition, which was shown by in vitro and in vivo measurements. Spectroscopical IR and Raman results showed the presence of bands whose intensity ratio can be related to the molar ratio R; in particular, the Raman I(962)/I(987) and the IR I(1035)/I(1125) intensity ratios were characterized as markers of the molar ratio. For these CaP compounds a nucleation model, which was based on the ability of HPO(2-)(4) ions to form strong H bonds with PO(3-)(4) ions, was proposed.

Development of arthritis and hypothyroidism during alpha-interferon therapy for chronic hepatitis C.

Alpha-interferon (alpha-IFN) therapy may induce, reveal or exacerbate various autoimmune-related disorders. The most common is the development of autoantibodies, while clinically overt autoimmune diseases are rare. We describe a 49-year-old woman who developed seronegative rheumatoid-like arthritis and autoimmune hypothyroidism after 7 months of human lymphoblastoid alpha-IFN therapy given for hepatitis C virus-related chronic active hepatitis (CAH-HCV). There was no family or personal history of autoimmune, thyroid or articular diseases. Our patient required continuous therapy for arthritis and hypothyroidism despite discontinuation of alpha-IFN. This suggests that alpha-IFN therapy may induce the contemporary appearance of two different persistent autoimmune-related diseases in the same patient. However, chronic HCV infection may play an important adjuvant role in the development of these diseases.

[Use of Doppler color ultrasonography in the microsurgical treatment of idiopathic varicocele]. / L'utilizzo dell'eco-color-Doppler nel trattamento microchirurgico del varicocele idiopatico.

The aim of this paper is to evaluate the role of Doppler and echo color-Doppler in the microsurgical treatment of varicocele. Since December 1993, 87 consecutive patients underwent microsurgical treatment of varicocele. The study demonstrates that Doppler and Color-flow-Duplex scanning provide an accurate, non-invasive method to identify the position of accessory spermatic veins, evaluating at the same time the flow characteristics of the microsurgical anastomosis.

[Pre- and postoperative color Doppler ultrasonography of myocutaneous flaps in reconstructive surgery of the breast]. / L'eco color-Doppler nello studio pre e postoperatorio dei lembi miocutanei in chirurgia ricostruttiva della mammella.

The authors present their experience in the study of blood supply of flaps. The machine used to study both pre and postoperatively the myocutaneous flap is the echo-color Doppler, a brand new device which has recently been used in investigating the morphology and the blood supply. The use of such method has provided us with the following information: detailed ultrasound structure of the skin, subcutaneous and muscular tissues of the examined flap; localization of vessels within the flap; size of the vessel in its whole length; quality and speed of both the arterial and venous flow. Such information is very important for the surgeon to plan in detail the reconstruction by knowing the blood supply of myocutaneous flap.