[The UniSpacer™: correcting varus malalignment in medial gonarthrosis. Preliminary results]. / UniSpacer™: corrección de la desalineación en varo en la gonartrosis medial. Resultados preliminares.

While options for operative treatment of leg axis varus malalignment in patients with medial gonarthrosis include several established procedures, such as unicompartmental knee arthroplasty (UKA), total knee arthroplasty (TKA), or high tibial osteotomy (HTO), so far there has been little focus on a less invasive option introduced more recently: the UniSpacer™ implant, a modern, self-centering, metallic interpositional device for the knee. This study evaluates whether alignment correction can be achieved by UniSpacer™ arthroplasty as well as alignment change in the first 5 postoperative years is evaluated. Antero-posterior long leg stance radiographs of 15 legs were digitally analyzed to assess alignment change: two relevant angles and the deviation of the mechanical axis of the leg were analyzed before and after surgery. Additionally, the change of the postoperative alignment was determined one and five years postoperatively. Analyzing the mechanical tibiofemoral angle, a significant leg axis correction was achieved, with a mean valgus change of 4.7±1.9°; a varus change occurred in the first postoperative year, while there was no significant further change of alignment seen five years after surgery. The UniSpacer™ corrects malalignment in patients with medial gonarthrosis; however, a likely postoperative change in alignment due to implant adaptation to the joint must be considered before implantation.

Stable isotope laser spectrometer for exploration of Mars.

On Earth, measurements of the ratios of stable carbon isotopes have provided much information about geological and biological processes. For example, fractionation of carbon occurs in biotic processes and the retention of a distinctive 2-4% contrast in 13C/12C between organic carbon and carbonates in rocks as old as 3.8 billion years constitutes some of the firmest evidence for the antiquity of life on the Earth. We have developed a prototype tunable diode Laser spectrometer which demonstrates the feasibility of making accurate in situ isotopic ratio measurements on Mars. This miniaturized instrument, with an optical path length of 10 cm, should be capable of making accurate 13C/12C and 15N/14N measurements. Gas samples for measurement are to be produced by pyrolysis using soil samples as small as 50 mg. Measurements of 13C/12C, 18O/16O and 15N/14N have been made to a precision of better than 0.1% and various other isotopes are feasible. This laser technique, which relies on the extremely narrow emission linewidth of tunable diode lasers (<0.001 cm(-1)) has favorable features in comparison to mass spectrometry, the standard method of accurate isotopic ratio measurement. The miniature instrument could be ready to deploy on the 2003 or other Mars lander missions.

Stable isotope analysis using tunable diode laser spectroscopy.

Measurements of ratios of stable isotopes are used in such diverse fields as petroleum prospecting, medical diagnostics, and planetary exploration. The narrow emission linewidth available from tunable diode lasers permits high-resolution infrared absorption measurements of closely spaced isotopic rovibrational lines. Our dual beam spectrometer uses the sweep integration technique in a spectral region where adjacent spectral lines are of approximately equal absorbance at the expected isotopic abundances. The experimental results reported here indicate that isotopic ratios of carbon in carbon dioxide can be measured to an accuracy of better than 0.4%. This laser spectroscopic technique offers an alternative to the mass spectrometric technique for in situ isotopic analysis in field studies, as well as flight and space applications.

Continuous monitoring of a changing sample by multiplex gas chromatography.

Multiplex gas chromatography (MGC) is a technique in which multiple samples may be introduced into a chromatographic system regardless of the elution time of the individual components. Although the output obtained from a MGC experiment is not directly interpretable, computational techniques can be used to obtain the chromatogram from the detector output data. This is done by calculating the impulse response function from the multiplexed output data.

Decay-associated emission spectra and other spectral evidence for the physical intercalation of 7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene into DNA.

A benzo[a]pyrene derivative, 7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene, forms physical complexes with DNA. The measured absorption spectrum of the hydrocarbon in the complex is shifted approximately 10 nm to the red and the fluorescence emission spectrum is red-shifted approximately 6 nm, characteristic of a physical intercalation complex. The decay-associated emission spectra of the hydrocarbon in the presence of DNA have been measured, thus providing a new technique to obtain information about the DNA binding sites. The decay-associated emission spectra of the free and bound hydrocarbons were obtained by deconvoluting the time-dependent emission at several wavelengths. Stern-Volmer plots with iodide and silver ions as quenchers suggest that at least one set of binding sites for the formation of a physical intercalation complex between the benzo[a]pyrene derivative and DNA is at guanine sites in the biopolymer.

Characterization of reversible, physical binding of benzo[a]pyrene derivatives to DNA.

The carcinogen, 7r,8t-dihydroxy-9t,10t-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene, and a number of its noncarcinogenic derivatives form a reversible physical complex with DNA in vitro. The absorbance spectrum of the hydrocarbon in the complex shifts approximately 10 nm to the red. Fluorescence from the physically bound hydrocarbon is significantly quenched, and binding levels are decreased by the addition of 1) spermine, 2) MgCl2, and 3) NaCl, with their relative effectiveness in reducing complex formation following the same order. Equilibrium data and the change in superhelicity of simian virus 40 DNA as a result of physical binding were used to calculate an unwinding angle for each of the benzo[a]pyrene derivatives of 13 degrees, while under the same conditions ethidium bromide unwound the duplex approximately 30 degrees. The weight of this evidence indicates that the carcinogen and its noncarcinogenic derivatives reversibly bind to DNA by intercalation of their planar aromatic system into the stacked base pairs of the duplex. The results of this and previous studies suggest that covalent binding of the carcinogenic derivative of benzo[a]pyrene to DNA is preceded by a physical intercalation step.

Problems in the disinfection of class 1 microbiology safety cabinets.

Microbiology safety cabinet disinfection procedures using formaldehyde have been tested. Tubercle bacilli were killed by concentrations of formaldehyde obtained by heating commercial formalin irrespective of whether the bacilli were in the cabinet free space or above the prefilters. However, Bacillus stearothermophilus spore papers for for the testing of low temperature steam/formaldehyde sterilisers were almost never sterilised and a strain of Staphylococcus epidermidis (NCTC 7944) showed a resistance intermediate between the B stearothermophilus spores and the tubercle bacilli. Tests using a vaccine strain of poliovirus type 3 indicated a considerable degree of resistance of the virus to the action of formaldehyde. No such resistance was demonstrated by vaccinia virus or echovirus 14. Chemical and biological evidence is presented which indicates that filter paper discs are an unsuitable carrier material for a challenge organism in testing the efficiency of any formaldehyde sterilising process. Recommendations are made towards developing a satisfactory test procedure.

Aryl hydrocarbon hydroxylase induction in mouse liver cells--relationship to position in the cell cycle.

The inducibility of aryl hydrocarbon hydroxylase (AHH) by benzo[a]-pyrene (BaP) has been studied in synchronously grown cultures of mouse liver cells. These cells (NMuLi cl 8) have low basal levels of AHH which can be induced greater than 100-fold by BaP. Cells were synchronized in G1(G0) by serum starvation and in S by release from serum starvation in combination with excess thymidine. When released from G1(G0) by replating at lower cell density in fresh medium with 20% serum, cells began entering S with a lag of 12 h. Addition of BaP (1 microgram/ml) 8 h before serum stimulation, at the time of stimulation or 7.5 h after stimulation all gave similar induction kinetics: the AHH activity peaked as the cells began entering S regardless of when the BaP was added. Cells blocked in various parts of S by excess thymidine were inducible for AHH activity as efficiently as cells moving through S and into G2. These results indicate that the inducibility of AHH is greater when cells are actively proliferating and may be a contributing factor to why growing cells are more sensitive to mutagenesis and transformation than quiescent cells.

Biochemical basis for the acquisition of resistance to benzo[a]pyrene in clones of mouse liver cells in culture.

In a Namru mouse liver epithelial cell strain designated NMuLi, aryl hydrocarbon hydroxylase (AHH) activity peaked at 12 h post-induction with 1 microgram/ml of benzo(a)pyrene (BaP) in both confluent and growing cells. Maximal levels of AHH activity were reached on day two post-plating. This induced activity was inhibited in vitro 78% by gassing the incubation mixture with carbon monoxide for 15 s, and inhibited 93% by addition of 40 microgram/ml of 7,8 benzoflavone(BF). Induced AHH levels were higher in epithelial clones that were sensitive to the toxicity of BaP than in resistant clones. The survival fraction of clones from NMuLi and of subclones derived from a sensitive clone of NMuLi after BaP treatment was a negative exponential function of the maximal induced AHH activity in the clones. One of the clones, NMuLi cl 8, was extremely susceptible to the toxic effects of BaP, the +/-(trans)-7alpha, 8beta-dihydroxy-7,8-dihydro-BaP(7,8-diol), and the (+/-)-7alpha, 8beta-dihydroxy-9beta, 10beta-epoxy-7,8,9,10-tetrahydro-BaP (diol-epoxide), known metabolites of BaP. The toxicity of BaP and the 7,8 diol to this clone was inhibited by BF, suggesting that these cells possessed an enzyme activity inhibitable by BF that could epoxidize BaP to the 7,8 oxide and then epoxidize the resultant 7,8 diol to the diol-epoxide. Another clone derived from NMuLi, clone 7, was relatively resistant to the toxic effects of BaP and the 7,8-diol, but still extremely susceptible to the toxic effects of the diol-epoxide. The slight toxicity to BaP in this clone was inhibited by BF, but the toxicity of the 7,8-diol to this clone was not inhibited by BF. A typical cytochrome P450 inhibitor, metyrapone, had no effect on the toxicity of BaP, the 7,8-diol, or the diol-epoxide to either clone 7 or clone 8. The results suggest that these liver cells possess two enzymes that play some role in polycyclic hydrocarbon-induced toxicity. Enzyme A, a BaP-inducible enzyme that is inhibitable by BF, efficiently metabolizes BaP to the 7,8-diol and the 7,8-diol to the diol-epoxide. It is responsible for most of the hydrocarbon toxicity. Enzyme B is not inhibitable by BF and metabolizes the 7,8-diol less efficiently to the diol-epoxide or efficiently to other, less toxic products.

Fatty acid requirements and temperature dependence of monooxygenase activity in rat liver microsomes.

The effect of variation in the microsomal membrane fatty acid composition on Arrhenius plot phase transition temperatures for p-nitroanisole O-demethylation and benzo[a]pyrene hydroxylation has been investigated. In liver microsomes from normal-dieted rats, p-nitroanisole O-demethylase activity has a break temperature at 24 degrees C, while that of benzo[a]pyrene hydroxylase occurs at 29 degrees C indicating that these two enzymes may exist in different patches of membrane. The microsomal membrane fatty acid composition was altered by starving rats for 48 h and then refeeding them a fat-free diet for 4 or 5 days. In microsomes having diet-altered fatty acid compositions, benzo[a]pyrene hydroxylase has a break temperature at 33 degrees C, a value higher than that observed in normal-dieted rats. This observation correlates with the increase in saturation observed in the diet-altered fatty acid composition and thus may correspond to a phase transition roughly dependent on the fatty acid melting point. Induced and basal levels of cytochrome P-450 and P-448 in animals having different microsomal fatty acid composition are reported. Phenobarbital-induced levels of p-nitroanisole O-demethylase in normal microsomes were six times higher than those in microsomes having diet-altered composition, whereas 3-methylcholanthrene-induced levels of benzo[a]pyrene hydroxylase were similar regardless of diet. The low level of p-nitroanisole O-demethylase activity in membranes with altered fatty acid compositions suggests that a particular type(s) of fatty acid was not present in sufficient quantity to permit the induction of maximal enzyme activity. Since the induced benzo[a]pyrene hydroxylase activity was the same regardless of diet, there was presumably sufficient quantities of the appropriate fatty acids present in the membrane for induction of this activity. Therefore, particular fatty acids may be necessary for the induction of maximal activity of particular enzymes in the mixed function monooxygenase system.

Photovoltages in suspensions of magnetically oriented chloroplasts.

The photovoltage of suspensions of magnetically oriented chloroplasts using polarized light of 680 nm has been measured. The magnitude of the photo e.m.f. depends on the polarization of the light and on its direction of propagation with respect to the oriented thylakoid planes. This photo e.m.f. is qualitatively attributed to the Dember effect which arises when inhomogeneous light absorption gives rise to a gradient of positive and negative charges along x , where x is the direction defined by the propagation vector of the light and which is also the direction joining the two electrodes. The photovoltage obtained with the planes of the oriented thylakoids parallel to x depends on the plane of polarization of the incident light and shows that (1) the magnitude of the photovoltage depends on the absorption coefficient (which itself is polarization dependent) and thus on the magnitude of the charge gradient produced by the inhomogeneously absorbed light, and (2) a charge gradient within the planes of the thylakoids can give rise to the macroscopic photovoltage. While our experimental observations are basically in agreement with those previously reported (Fowler, C.F. and Kok, B.(1974) Biochim. Biophys. Acta 357, 308-318 and Witt, H.T. and Zickler, A. (1973) FEBS Lett. 37, 307-310) for unoriented chloroplasts, their interpretation of the origin of this effect in terms of a transmembrane potential must be modified in view of our results obtained with oriented chloroplasts. The macroscopically observed photovoltage of oriented chloroplasts is due to the creation of charge gradients either parallel or perpendicular to the thylakoid planes by a flash of light, the diffusion of these charges and to differences in the mobilities of the negative and positive charges. This interpretation in terms of the Dember effect is completely consistent with the existence of a transmembrane electric field as proposed by Fowler and Kok, as well as by Witt and Zickler. However, macroscopic measurements of the photovoltage using either oriented or unoriented chloroplast suspensions cannot prove that a transmembrane voltage exists, as previously claimed.

Anisotropy of photosynthetic membranes and the degree of fluorescence polarization.

The degree of fluorescence polarization, P, of unoriented and magnetically oriented spinach chloroplasts as a function of excitation (400-680 nm) and emission wavelengths (675-750 nm) is reported. For unoriented chloroplasts P can be divided into two contributions, PIN and PAN. The latter arises from the optical anisotropy of the membranes which is due to the orientation with respect to the membrane plane of pigment molecules in vivo. The intrinsic polarization PIN, which reflects the energy transfer between different pigment molecules and their degree of mutual orientation, can be measured unambiguously only if (1) oriented membranes are used and the fluorescence is viewed along a direction normal to the membrane planes, and (2) the excitation is confined to the Qy (approximately 660-680 nm) absorption band of chlorophyll in vivo. With 670-680 nm excitation, values of P using unoriented chloroplasts can be as high as +14%, mostly reflecting the orientational anisotropy of the pigments. Using oriented chloroplasts PIN is shown to be +5+/-1%. The excitation wavelength dependence studies of PIN indicate that the carotenoid and chlorophyll Qy transition moments tends to be partially oriented with respect to each other on a local level (within a given photosynthetic unit or its immediate neighbors).