The isoflavones genistein and daidzein increase hepatic concentration of thyroid hormones and affect cholesterol metabolism in middle-aged male rats.

We examined whether isoflavones interfere with thyroid homeostasis, increase hepatic thyroid hormone concentrations and affect cholesterol metabolism in middle-aged (MA) male rats. Thirteen-month-old Wistar rats were injected subcutaneously with 35 mg/kg b.w./day of genistein, daidzein or vehicle (controls) for four weeks. Hepatic Dio1 gene expression was up-regulated by 70% (p < 0.001 for both) and Dio1 enzyme activity increased by 64% after genistein (p < 0.001) and 73% after daidzein treatment (p < 0.0001). Hepatic T3 was 75% higher (p < 0.05 for both), while T4 increased only after genistein treatment. Serum T4 concentrations were 31% lower in genistein- and 49% lower in dadzein-treated rats (p < 0.001 for both) compared with controls. Hepatic Cyp7a1 gene expression was up-regulated by 40% after genistein and 32% after daidzein treatment (p < 0.05 for both), in agreement with a 7α-hydroxycholesterol increase of 50% (p < 0.01) and 88% (p < 0.001), respectively. Serum 24- and 27-hydroxycholesterol were 30% lower (p < 0.05 for both), while only 24-hydroxycholesterol was decreased in the liver by 45% after genistein (p < 0.05) and 39% (p < 0.01) after dadzein treatment. Serum concentration of the cholesterol precursor desmosterol was 32% (p < 0.05) lower only after dadzein treatment alone, while both isoflavones elevated this parameter in the liver by 45% (p < 0.01). In conclusion, isoflavones increased T3 availability in the liver of MA males, despite decreasing serum T4. Hepatic increase of T3 possibly contributes to activation of the neutral pathway of cholesterol degradation into bile acids in the liver. While isoflavones obviously have the potential to trigger multiple mechanisms involved in cholesterol metabolism and oxysterol production, they failed to induce any hypocholesterolemic effect.

Antimicrobial plant metabolites: structural diversity and mechanism of action.

Microbial infectious diseases continue to be one of the leading causes of morbidity and mortality. It has been estimated that microbial species comprise about 60% of the Earth's biomass. This, together with the fact that their genetic, metabolic and physiological diversity is extraordinary, makes them a major threat to the health and development of populations across the world. Widespread antibiotic resistance, the emergence of new pathogens in addition to the resurgence of old ones, and the lack of effective new therapeutics exacerbate the problems. Thus, the need to discover and develop new antimicrobial agents is critical to improve mankind's future health. Plant secondary metabolites (PSMs) offer particular promise in this sense. Plant Kingdom could be considered a rich source of the most diverse structures (e.g. there are more than 12,000 known alkaloids, more than 8,000 phenolic compounds and over 25,000 different terpenoids), many of which were proven to possess strong antimicrobial properties (e.g. thymol, eurabienol, etc.). In many instances, PSMs can be easily isolated from the plant matrix, either in pure state or in the form of mixtures of chemically related compounds. What is also important is that the development of bacterial resistance toward natural plant products (that are generally regarded as eco-friendly) has been thus far documented in a very limited number of cases (e.g. for reserpine). Having all of the mentioned advantages of PSMs as potential antimicrobials in mind, a major question arises: why is it that there are still no commercially available or commonly used antibiotics of plant origin? This review tries to give a critical answer to this question by considering potential mechanisms of antimicrobial action of PSMs (inhibition of cell wall or protein synthesis, inducing leakage from the cells by tampering with the function of the membranes, interfering with intermediary metabolisms or DNA/RNA synthesis/function), as well as their physical and chemical properties (e.g. hydrophilicity/lipophilicity, chemical stability). To address the possible synergistic/antagonistic effects between PSMs and with standard antibiotics, special attention has been given to the antimicrobial activity of PSM-mixtures (e.g. essential oils, plant extracts). Moreover, possible ways of overcoming some of PSMs molecular limitations in respect to their usage as potential antibiotics were also discussed (e.g. derivatization that would enable fine tuning of certain molecular characteristics).

Antistaphylococcal activity of Inula helenium L. root essential oil: eudesmane sesquiterpene lactones induce cell membrane damage.

The purpose of this study was to investigate the inhibitory/bactericidal activity and cell membrane effects of the hydrodistilled essential oil of Inula helenium L. roots against Staphylococcus aureus. Additionally, detailed chemical investigation was done in order to pinpoint the most active oil constituents and also the parts of these molecules responsible for their antimicrobial effect. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined using the broth microdilution method. The membrane-active nature of this oil was investigated by measuring the culture turbidity, leakage of phosphates, and 260-nm-absorbing material, together with lysis of the exposed cells. Finally, the effect of the oil on the cells was visualized using scanning electron microscopy (SEM). The chemical composition of the essential oil was analyzed using gas chromatography-mass spectrometry (GC-MS) and preparative medium-pressure liquid chromatography (MPLC). Chemical modification of the oil was performed using catalytic hydrogenation (H(2), Pd/C) and reduction with NaBH(4). The MIC and MBC values were 0.01 µl mL(-1) and 0.02 µl mL(-1), respectively. Membrane damage was demonstrated through increased permeability (phosphates and nucleic acid leakage), followed by lysis of the exposed cells, captured on SEM images. The most active constituents were alantolactone, isoalantolactone, and diplophyllin. The essential oil showed very potent antistaphylococcal activity, with obvious membrane-damaging effects. Sesquiterpene lactones were found to be the most active principles of the oil, whose eudesmane core olefinic bonds, along with the α,ß-methylene-lactone ring, are essential structural parts responsible for the exhibited antimicrobial activity.

Development and validation of liquid chromatography tandem mass spectrometry methods for the determination of gentamicin, lincomycin, and spectinomycin in the presence of their impurities in pharmaceutical formulations.

Liquid chromatography with tandem mass spectrometry (LC/MS/MS) methods for the determination of gentamicin, lincomycin and spectinomycin in the presence of their impurities were developed and tested. Chromatographic separations were achieved using gradient elution on a C18 column. All components were ionized by positive-ion electrospray and detected by multi reaction monitoring (MRM) with an LC-tandem mass spectrometer. Calibration curves were linear with correlation coefficients better than 0.99. The developed method for the determination of gentamicin provides complete base line separation of components C1, C1a, C2, C2a and C2b mentioned in the European and British Pharmacopoeias. The second developed method makes possible a simultaneous analysis of the active compounds of both lincomycin and spectinomycin. Additionally, all impurities defined in the pharmacopoeias for all three active components were determined and their identities confirmed. The methods were tested in routine quality control analysis.

Antimicrobial synergism and antagonism of salicylaldehyde in Filipendula vulgaris essential oil.

The leaf essential oil of Filipendula vulgaris, consisting mainly of salicylaldehyde (68.6%), was screened for its antimicrobial activity by the disk diffusion and microdilution broth assays. The essential oil remarkably inhibited the growth of all of the tested bacteria and fungi. It seems that the antimicrobial nature of F. vulgaris essential oil can be attributed to the synergistic interactions of the compounds constituting the oil rather than to the presence of a single inhibitory agent. A synergy in salicylaldehyde/linalool mixtures was observed with a maximum interaction situated in the range between 60:40 and 80:20 (mol ratio). At this concentration range (at a dose of 1.7 microg/disk) no microbial growth was observed while the respective pure compounds, at the corresponding quantities, are shown to be dramatically less active. The MIC value for the 60:40 mixture was determined to be less that 0.009 mg/ml. In addition, an antagonistic relationship between salicylaldehyde and methyl salicylate was established. The maximum (negative) interaction was shown to correspond approximately to the mixture at the 40:60 (methyl salicylate/salicylaldehyde) mol ratio resulting in the complete loss of activity at the investigated dose.

[Rigid-fix pins fixation and LCA reconstruction with semitendinosus/gracilis autograft].

ACL reconstruction techniques highly developed during the decades and biomechanical tests for hamstrings graft showed much in advance loading and stiffness comparing BPTB grafting. We have made joint analysis of 30 patients with semitendinosus / gracilis ACL reconstruction. Choice for proximal fixation was Mitek Rigid-fix Pins technique and bioactive screws for distal fixation; in 18 cases bioactive screws were used for femoral and tibial graft fixation. Lysholm score and knee clinical tests were parameters used for outcome evaluation. Follow up averaged 6 months with assessment after 1, 3, 6 and 12 months. Our results showed excellent postoperative Lysholm score in 85% and very good in 15% of cases. Postoperative Lachman was 1+ in 15% and postoperative pivot shift was 0%. Full rehabilitation achieved 85% and with modifications in everyday activities 15% of patients. Donor localization morbidity was markedly reduced and we have no complications. On the basis of the improved results in graft compression as well as stability with this technique, applied method is highly recommended for the reconstruction in ACL deficient knees.

[External fixation of war injuries of the proximal femur].

Seventeen patients with open fractures of the upper third of the femur were treated using a pelvifemoral external fixation device. All of them had grade III open fractures resulting from high-velocity missile and explosive injuries with massive foreign body contamination. Sciatic nerve injury was present in five (29.4%); abdominal viscera and thoracic wall injuries were present in two patients (11.8%). There were no major arterial injuries. Full weight bearing was allowed after clinical and radiological bone healing (average 11.5 months). Chronic osteitis with fistula and sequestra developed in two (11.8%) patients. There were no nonunions and no refractures. Minor painless limitation of hip motion persisted in all patients. Upper-third femoral open fractures due to firearms are a unique type of open fractures. They are usually highly comminuted; therefore, stable fixation is difficult or impossible to achieve using external fixation with transfixation of the fracture site. On the other hand, the risk of infection is high following intramedullary nailing. Pelvifemoral external fixation allows adequate management of the soft tissue wounds, provides stable bone fixation and allows early patient mobilization.

[ACL reconstruction with BTB graft--unusual evolution of knee arthrofibrosis case report].

Knee arthrofibrosis, which usually occurs after trauma or surgery, can inhibit joint biomechanics. An elaborated interaction of growth factors and other inflammatory mediators initiates and coordinates this deleterious tissue proliferation. Knowledge of risk factors can aid clinicians in helping patients avoid knee arthrofibrosis. Once the condition is present, a history and examination are imperative to institute the most appropriate treatment regimen. Nonoperative measures can be used as therapy, though surgery is often necessary for optimal results. We have analyzed problems in patient with uncommon evolution of knee arthrofibrosis following ACL recontruction with BTB autograft.

Gemeprost versus misoprostol for cervical priming before first-trimester abortion: a randomized controlled trial.

OBJECTIVE: To compare the efficacy of 400 microg of misoprostol with that of 1 mg of gemeprost as cervical priming agents when administered vaginally 3 to 4 hours before first-trimester vacuum aspiration abortion. METHODS: In a prospective controlled trial 90 nulliparous women who requested termination of pregnancy before 12 weeks' gestation were randomized to receive vaginally either misoprostol or gemeprost for cervical priming. The force to dilate the cervix was measured by the use of a cervical tonometer connected to Hegar dilators from 3 to 10 mm. The main outcome measures were baseline cervical dilation; the peak force to dilate the cervix at 8, 9, and 10 mm; and the cumulative force to dilate the cervix to 10 mm. RESULTS: Baseline cervical dilation did not differ significantly between the women who received misoprostol and those who were treated with gemeprost. Neither the peak force required to dilate the cervix at 8, 9, and 10 mm nor the cumulative force to dilate the cervix to 10 mm showed any significant difference between the two groups. CONCLUSION: Vaginally administered misoprostol (400 microg) is as effective as gemeprost (1 mg) for cervical priming 3 to 4 hours before surgical termination of first-trimester pregnancies.

Effects of estradiol and calcium on gonadotrophic cells in middle-aged female rats.

The effects of multiple treatment with estradiol dipropionate (EDP) or calcium glucoheptonate (Ca) or a combination of the two on gonadotrophic cells in the pituitary pars distalis of middle-aged female rats were examined. The animals were treated daily for two weeks with EDP (0.625 mg i.p./kg body weight) or Ca (11.4 mg/kg body weight) or EDP + Ca. Luteinising (LH) and follicle stimulating hormone (FSH)-producing cells were examined by immunohistochemistry using antisera to the specific (beta) beta-subunits of LH and FSH and a peroxidase-anti-peroxidase immunohistochemical procedure. Plasma levels of FSH and LH were measured by radio-immune assay. A stereological method for determining morphometric parameters in immunopositive FSH and LH cells was used. The number of gonadotrophs per unit area (mm2), their cellular volume and relative volume densities, as well as plasma levels of FSH and LH, were decreased in all treated females in comparison with the controls. The most significant decrease of these parameters was observed in EDP-treated animals. Such changes were also expressed in Ca-treated animals, but the alterations were less distinct. These results demonstrate that multiple EDP or Ca application to middle-aged female rats is able to inhibit, directly or indirectly, the morphofunctional state of gonadotrophic cells in the pituitary pars distalis.

Effects of somatostatins on gonadotrophic cells in female rats.

The effects of intracerebroventricular application of SRIF-14 and SRIF-28 on pituitary gonadotrophic cells (FSH and LH) were examined using immunocytochemical and morphometrical methods in adult female Wistar rats. FSH- and LH-producing cells were studied using the peroxidase-antiperoxidase (PAP) immunohistochemical procedure. Morphometry and stereology were used to evaluate changes in the number, volume densities and relative volume densities of LH- and FSH-immunopositive cells. In females treated with SRIF-14 or SRIF-28, the gonadotrophs were smaller, often pycnotic and more intensely stained. The number of LH-positive cells per unit area (mm2) was significantly decreased in both somatostatin-treated groups, while the number of FSH-positive cells was similar to that in the controls. Volume densities of perykarya of LH- and FSH-positive cells were decreased in all treated females, but extremely different in LH-positive cells after SRIF-14 administration. The relative volume density of LH cells was significantly decreased in both somatostatin-treated groups, while immunopositive FSH cells were not significantly decreased compared with the controls. It can be concluded that centrally applied somatostatins lead to changes in the immunocytochemical and morphometric properties of LH cells, while there is no significant effect on FSH cells.