Variation in human vertebral body strength for vertebral body samples from different locations in segments L1-L5.

BACKGROUND: The human spine, in particular the lumbar spine, is subject to significant compressive and bending stresses, which affect the structure of the bone tissue of the vertebrae. The more heterogeneous the structure of the spongy bone tissue, the less resistant the whole vertebral body. It is therefore necessary to establish variations in bone strength parameters within one particular vertebral body. METHODS: The research material comprised human L1-L5 lumbar vertebrae sampled from 15 donors aged 29-35. A total of 975 samples prepared from the collected material were subjected to compressive and bending strength tests. The samples for the tests were collected from carefully selected locations in order to discover the strength properties of various parts of the vertebral body. FINDINGS: In the case of sample 2 (located in the posterior part of the vertebra, at mid-height) the stress values were the lowest and there were statistically significant differences compared to other samples. Moreover the value of compressive force in this case was lower for vertebrae with higher numbers. Top and bottom samples demonstrated statistically significant higher mean values of destructive stress. In terms of the bending strength test, the mean value of destructive stress in all lumbar vertebrae for all samples increased for vertebrae with higher numbers. INTERPRETATION: The spongy tissue in healthy vertebral bodies has a very heterogeneous structure. This may be due to the presence of the nutrient canal and the arc structure allowing more springy movement and improved transfer of loads by the vertebral body.

Impact of empiric weight-based vancomycin dosing on nephrotoxicity and mortality in geriatric patients with methicillin-resistant Staphylococcus aureus bacteraemia.

WHAT IS KNOWN AND OBJECTIVE: Few studies have evaluated the effect of vancomycin dosing on the health outcomes in geriatric patients. Data are needed to determine whether higher vancomycin dosing strategies are more effective in geriatric patients and/or lead to excessive rates of adverse events. METHODS: This study used a subset of patients aged ≥65 years from a multicentre, retrospective, cohort study of methicillin-resistant Staphylococcus aureus (MRSA) bacteraemia. Patients received ≥ 48 h of empiric vancomycin between 1 July 2002 and 30 June 2008. We compared the incidence of nephrotoxicity and in-hospital mortality in patients who received guideline-recommended dosing (at least 15 mg/kg/dose) to patients who received lower dosing. Multivariable generalized mixed-effect models were constructed to determine independent risk factors for nephrotoxicity and in-hospital mortality. RESULTS AND DISCUSSION: Half of the cohort (46% of 92 patients) received guideline-recommended dosing. Empiric use of weight-based dosing did increase the percentage of patients achieving a vancomycin trough ≥ 15 mg/L (57% vs. 42%). Nephrotoxicity occurred in 32% of patients and 26% died during their hospitalization. Guideline-recommended dosing was not associated with significant changes in nephrotoxicity (OR 1·13; 95% CI 0·40-3·19) or in-hospital mortality (OR 1·14; 95% CI 0·41-3·18) in the multivariable analysis. WHAT IS NEW AND CONCLUSION: In this study of geriatric patients, guideline-recommended dosing was not associated with significant changes in nephrotoxicity or mortality. As 40% of the patients who received guideline-recommended dosing failed to achieve a target vancomycin trough of ≥ 15 mg/L, future studies should focus on dosing strategies to increase target attainment rate.

Cell survival and chromosomal aberrations in CHO-K1 cells irradiated by carbon ions.

Chinese hamster ovary CHO-K1 cells were exposed to high LET (12)C-beam (LET: 830 keV/microm) in the dose range of 0-6 Gy and to (60)Co irradiation and the RBE value was obtained. Effects of (12)C-beam exposure on cell survival and chromosomal aberrations were calculated. The chromosomal aberration data were fitted with linear equation. The distribution of aberration in cells was examined with a standard u-test and used to evaluate the data according to Poisson probabilities. The variance to the mean ratio sigma(2)/Y and the dispersion index (u) were determined. Overdispersion was significant (p<0.05) when the value of u exceeded 1.96.

Evaluation of the genotoxic and antioxidant effects of two novel feed additives (ethoxyquin complexes with flavonoids) by the comet assay and micronucleus test.

The complexes of antioxidant ethoxyquin (1,2-dihydro-6-ethoxy-2,2,4-trimethylquinoline; EQ) with rutin or quercetin (EQ-R and EQ-Q, respectively) were studied in human lymphocytes for genotoxic and antioxidant activities with the use of the comet assay and micronucleus test. The study was undertaken to search for new potential antioxidants, and was motivated by reports of unfavourable side-effects observed in animals fed with feeds containing EQ, which is allowed up to 150 mg kg(-1) (0.015%) in complete animal feed. It was shown that EQ-R induced DNA damage in human lymphocytes when used at all the concentrations studied (1-25 microM), while after EQ-Q treatment, the genotoxic effect was observed mainly after higher doses (10 and 25 microM). An increase in the number of micronuclei was observed only for EQ-Q after a dose of 50 microM. The studied compounds decreased the degree of DNA damage induced by hydrogen peroxide (10 microM) in the comet assay. The results obtained in both tests showed that the antioxidant activity of EQ-Q was comparable with that of EQ, so further detailed studies are necessary to estimate its possible usefulness as a feed preservative.

Two-dimensional assembly and local redox-activity of molecular hybrid structures in an electrochemical environment.

The self-assembly and redox-properties of two viologen derivatives, N-hexyl-N'-(6-thiohexyl)-4,4'-bipyridinium bromide (HS-6V6-H) and N,N'-bis(6-thiohexyl)-4,4'-bipyridinium bromide (HS-6V6-SH), immobilized on Au(lll)-(1 x 1) macro-electrodes were investigated by cyclic voltammetry, surface enhanced infrared spectroscopy (SEIRAS) and in situ scanning tunneling microscopy (STM). Depending on the assembly conditions one could distinguish three different types of adlayers for both viologens: a low coverage disordered and an ordered "striped" phase of flat oriented molecules as well as a high coverage monolayer composed of tilted viologen moieties. Both molecules, HS-6V6-H and HS-6V6-SH, were successfully immobilized on Au(poly) nano-electrodes, which gave a well-defined redox-response in the lower pA-current range. An in situ STM configuration was employed to explore electron transport properties of single molecule junctions Au(T)/HS-6V6-SH(HS-6V6-H)/Au(S). The observed sigmoidal potential dependence, measured at variable substrate potential E(S) and at constant bias voltage (E(T) - E(S)), was attributed to electronic structure changes of the viologen moiety during the one-electron reduction/re-oxidation process V2+ < -- > V+*. Tunneling experiments in asymmetric, STM-based junctions Au(T)-S-6V6-H/Au(S) revealed current (i(T))-voltage (E(T)) curves with a maximum located at the equilibrium potential of the redox-process V2+ < -- > V+*. The experimental i(T)--E(T) characteristics of the HS-6V6-H-modified tunneling junction were tentatively attributed to a sequential two-step electron transfer mechanism.

Induction of chromosome aberrations in cultured human lymphocytes treated with ethoxyquin.

The chromosomal aberration test was employed to investigate the effect in vitro of a known antioxidant and food preservative, ethoxyquin (EQ, 1,2-dihydro-6-ethoxy-2,2,4-trimethylquinoline) on human chromosomes. The studies were undertaken because there are no published in vitro data on genotoxicity of EQ in mammalian cells and there are many reports pointing out that it may be harmful to animals and human beings. Lymphocytes obtained from three healthy donors were incubated with EQ (0.01-0.5mM) both with and without metabolic activation. Stability studies performed by HPLC analysis showed that EQ was stable under the conditions of the lymphocyte cultures. The results of the chromosome aberration assay showed that EQ induces chromosome aberrations: gaps and breaks as well as dicentrics and atypical translocation chromosomes.

Altered expression of nuclear non-histone protein (p44/46) in different stages of B-chronic lymphocytic leukemia.

Our previous data have shown some differences in electrophoretic characteristics of proteins from cellular fractions (nuclear, mitochondrial, microsomal and cytosolic) isolated from peripheral blood mononuclear cells of B-cell chronic lymphocytic leukemia (B-CLL), acute lymphoblastic leukemia (ALL) patients and healthy donors. The main differences were found in electrophoretic patterns of nuclear proteins from normal and leukemia cells, especially in the nuclear mass regions of 36-52, 58-85, and 120-180 kDa. Electrophoretically-specific nuclear non-histone protein in the molecular mass zone 44/46 kDa of cells obtained from the peripheral blood of a B-CLL patient was used to produce rabbit polyclonal antiserum. SDS-polyacrylamide gel electrophoresis as well as immunological techniques (Western blot and immunocytochemistry) indicate that the nuclear protein with a molecular mass of 44/46 kDa is specifically expressed in mononuclear cells from B-CLL patients. The expression of this particular nuclear protein seems to correlate with the progression of the leukemia.

Modification of non-protein thiols contents in transgenic tobacco plants producing bacterial enzymes of cysteine biosynthesis pathway.

Conditions of achieving the maximal accumulation of sulfhydryl metabolites in the leaves of tobacco were explored. Simultaneous production of bacterial O-acetylserine (thiol)-lyase and serine acetyltransferase resulted in the increased thiols contents as compared to single transformants and controls. However, leaf discs feeding experiments differently affected thiols concentration in different plant groups and suggested that the most promising strategy to obtain plants with a high level of non-protein thiol-containing compounds might be sulfate feeding to plants overproducing serine acetyltransferase.

Synthesis and antimicrobial activity of (E)-acetoxystilbenes and alpha,alpha'-dibromoacetoxybibenzyls.

The synthesis of 13 new (E)-acetoxystilbenes and alpha,alpha'-dibromoacetoxybibenzyls and their antimicrobial activity are reported. The results of microbiological screening of 17 (E)-stilbenols and (E)-acetoxystilbenes, unknown in the literature, have also been discussed. In particular, coumpounds 1c, 1g, 2a, 3a, 3b, 4a, 6a, 6b showed good antibacterial activity against Staphylococcus aureus and 1c also against Bacillus subtilis.

Increased resistance to oxidative stress in transgenic tobacco plants overexpressing bacterial serine acetyltransferase.

Plant expression cassettes containing the Escherichia coli cysE gene alleles (encoding SAT) were constructed. After the Agrobacterium-mediated transformation of tobacco, we identified stable transformed plants containing several-fold higher SAT activity in comparison to the control plant. Determination of non-protein thiol contents indicated two- to threefold higher cysteine and glutathione levels in some of these transgenic plants. The maximal elevation of the cysteine level was about fourfold while that of GSH was about twofold higher than in the controls. The most striking physiological consequence of the modification of sulfur metabolite levels in the transgenic plants, however, was their several-fold increased resistance to oxidative stress generated by exogenous hydrogen peroxide.

In vitro maturation of human preovulatory oocytes reconstructed by germinal vesicle transfer.

OBJECTIVE: To describe a micromanipulation-electrofusion procedure for transferring germinal vesicles (GVs) between immature human oocytes. DESIGN: Pilot study to assess oocyte maturation after an invasive micromanipulation procedure. SETTING: Research laboratory at a university medical center. PATIENT(S): Immature oocytes were discarded from intracytoplasmic sperm injection (ICSI)-IVF cycles of patients 23-48 years of age. INTERVENTION(S): Initially, GV removal and transfer were performed on the same oocyte; these "self-reconstructed" oocytes were then cultured in vitro for up to 50 hours and examined periodically for maturation as judged by the extrusion of the first polar body. In a second study, GVs from oocytes of "old" patients (>38 years old) were successfully transferred into enucleated immature oocytes of "young" patients (<31 years old). MAIN OUTCOME MEASURE(S): Extrusion of the first polar body was monitored in "reconstructed" and control oocytes; karyotypes also were analyzed at meiosis II. RESULT(S): From 48 oocytes from old patients, 12 GVs were successfully removed, transferred, and fused into previously enucleated oocytes from young patients. After in vitro culture, 7 of these "reconstructed" oocytes matured to meiosis II, a maturation rate not significantly different from that observed in nonmanipulated controls. A normal, second meiotic metaphase chromosome complement was observed in 4 of 5 reconstructed oocytes. CONCLUSION(S): Normal meiosis can occur after the transfer of a GV into an enucleated host oocyte. Germinal vesicle transfer may be a valuable research procedure that generates cell models to characterize the cytoplasmic-nuclear interplay for cell cycle regulation, maturation, and fertilization in the human oocyte; it also may be a potentially attractive alternative to oocyte donation.

Preimplantation genetic diagnosis of human embryos for Marfan's syndrome.

PURPOSE: Single-cell nested polymerase chain reaction (PCR) and Ddel endonuclease digestion were used to detect the presence of a Marfan's syndrome mutation in human preimplantation embryos derived from in vitro fertilization (IVF). These procedures were conducted to eliminate the possibility of transmission of the affected allele from the father to his offspring. The mutation on chromosome 15 is transmitted as an autosomal dominant trait, and the chance of having a child affected with the disease is 50%. METHODS: A couple presented to the Program for In Vitro Fertilization, Reproductive Surgery and Infertility for preimplantation genetic diagnosis. IVF was performed and embryo biopsy was done on day 3 embryos. Single blastomeres were removed from embryos and subjected to nested PCR analysis and endonuclease digestion to detect a Marfan's syndrome mutation located on chromosome 15 inherited from the father. RESULTS: Thirteen oocytes were injected with spermatozoa using intracytoplasmic sperm injection, and nine fertilized normally. Following embryo biopsy and polymerase chain reaction amplification-Ddel endonuclease digestion, five embryos were detected that were positive for the mutation. The four non-affected embryos were transferred to the uterus, resulting in a healthy and normal ongoing pregnancy.