Adolescent idiopathic scoliosis (AIS) treated with arthrodesis and posterior titanium instrumentation: 8 to 12 years follow up without late infection

Background

There are no data in the peer-reviewed literature regarding long term results in patients treated for AIS with a posterior titanium instrumentation. Therefore we assessed the outcome in 50 patients treated by titanium implant.

Methods

A total of 50 patients with a mean age of 16.6 years were treated. In all patients, titanium hooks and pedicle screws were used in combination. The demographic data and the pre- and post-operative radiographs of all 50 patients were re-examined, and 49 of the 50 patients (98%) attended a radiological and clinical follow up-examination on average 10.1 years post-operatively. The clinical results were recorded by means of the SRS 24 questionnaire.

Results

In the frontal plane, the mean pre-operative thoracic and lumbar curve had been 62.4° and 43.5° respectively, post-operatively the curves were reduced to 26.9° and 16.3°, resulting in a correction rate of 56.9% for thoracic and 62.5% for lumbar curve. At the follow up-evaluation, the Cobb angle of the thoracic and lumbar curve was 31.0° and 21.3° respectively, giving a final correction rate of 50.3% for thoracic, and 51.0% for lumbar curve. 7 of the 50 patients (14.3%) had undergo revision surgery for complications, but complete implant removal was necessary in only one case. Analysis of the SRS 24 questionnaire showed an average score of 95.8 points.

Conclusion

Posterior titanium instrumentation is a safe and effective procedure in the surgical correction of AIS. In this retrospective study with small patient number, it shows favourable long-term results; in particular, the loss of correction is low, no late infection occurred and there was a very high survival rate of the implant itself.

     

Immediate effects of scoliosis-specific corrective exercises on the Cobb angle after one week and after one year of practice

Background

We are unaware of any studies describing the immediate effects of scoliosis-specific exercises on the Cobb angle measured by radiograph. This study aimed to describe the differences between radiographs obtained with and without corrective exercises after initial training and after one year.

Methods

A female with adolescent idiopathic scoliosis was first seen at age 13 years, 0 months with a Risser 0. She had a 43^o left lumbar, 15^o right thoracic curve. She was seen again after 6, 18 and 30 months and performed exercises from 18 to 30 months. She performed Barcelona Scoliosis Physical Therapy School (BSPTS) exercises for a four-curve type (lumbar dominant with pelvis deviation to the lumbar concave side). At 18 and 30 months, x-rays were obtained with and without performing corrective exercises.

Results

At 6 months, her lumbar and thoracic curves measured 41^o and 28^o, respectively. At 18 months, her lumbar and thoracic curves measured 47 ^o and 30^o, respectively. Also at 18 months, immediately after her x-ray in the relaxed standing position, she performed her corrective exercises in standing with arms lowered for a second x-ray. Her lumbar and thoracic curves remained similar and measured 43^o and 32^o, respectively. At 30 months, she performed unsolicited corrective exercises during the x-ray. Her lumbar and thoracic curves measured 26^o and 41^o, respectively. Another x-ray in the relaxed position revealed lumbar and thoracic curves measuring 39^o and 35^o, respectively. The immediate effect of corrective exercises after a year of training was a 33 % improvement at the lumbar spine compared to only a 9 % improvement the previous year.

Conclusion

After initial training, corrective exercises during a standing x-ray did not significantly improve the Cobb angle for the major lumbar curve compared to the relaxed standing x-ray. However, a year after performing exercises, unsolicited corrective exercises resulted in a significantly improved Cobb angle compared to relaxed standing for the curve primarily targeted by the exercise program. Improved exercise ability and spinal flexibility may have contributed to the improved Cobb angle.

     

A novel ethanol-tolerant laccase,Tvlac, from <Emphasis Type="Italic">Trametes versicolor</Emphasis>

Objectives

To produce and characterize novel laccases with ethanol tolerance from Trametes versicolor using agriculture by-products as energy source.

Results

Trametes versicolor 1017 produces two laccase isoenzymes with a total activity of 10 U ml^?1 within 8 days when using wheat bran and peanut powder as energy sources in liquid culture medium. A novel isoenzyme, named Tvlac, was identified, purified and characterized. Its optimum pH and temperature were from 4.5 to 5 and 55 to 60 °C, respectively. Its activity was stimulated by ethanol at 10 % (v/v) which increased the V ₀.

Conclusions

The biochemical properties of Tvlac substantiate the potential of this enzyme for applications under an aqueous ethanol mixture environment.

     

A high effective NADH-ferricyanide dehydrogenase coupled with laccase for NAD<Superscript>+</Superscript> regeneration

Objectives

To find an efficient and cheap system for NAD⁺ regeneration

Results

A NADH-ferricyanide dehydrogenase was obtained from an isolate of Escherichia coli. Optimal activity of the NADH dehydrogenase was at 45 °C and pH 7.5, with a K _m value for NADH of 10 μM. By combining the NADH dehydrogenase, potassium ferricyanide and laccase, a bi-enzyme system for NAD⁺ regeneration was established. The system is attractive in that the O₂ consumed by laccase is from air and the sole byproduct of the reaction is water. During the reaction process, 10 mM NAD⁺ was transformed from NADH in less than 2 h under the condition of 0.5 U NADH dehydrogenase, 0.5 U laccase, 0.1 mM potassium ferricyanide at pH 5.6, 30 °C

Conclusion

The bi-enzyme system employed the NADH-ferricyanide dehydrogenase and laccase as catalysts, and potassium ferricyanide as redox mediator, is a promising alternative for NAD⁺ regeneration.

     

The effect of scoliotic deformity on spine kinematics in adolescents

Background

While adolescent idiopathic scoliosis (AIS) produces well characterized deformation in spinal form, the effect on spinal function, namely mobility, is not well known. Better understanding of scoliotic spinal mobility could yield better treatment targets and diagnoses. The purpose of this study was to characterize the spinal mobility differences due to AIS. It was hypothesized that the AIS group would exhibit reduced mobility compared to the typical adolescent (TA) group.

Methods

Eleven adolescents with right thoracic AIS, apices T6-T10, and eleven age- and gender-matched TAs moved to their maximum bent position in sagittal and coronal plane bending tasks. A Trakstar (Ascension Technologies Burlington, VT) was used to collect position data. The study was approved by the local IRB. Using MATLAB (MathWorks, Natick, MA) normalized segmental angles were calculated for upper thoracic (UT) from T1-T3, mid thoracic (MT) from T3-T6, lower thoracic (LT) from T6-T10, thoracolumbar (TL) from T10-L1, upper lumbar (UL) from L1-L3, and thoracic from T1-L1 by subtracting the standing position from the maximum bent position and dividing by number of motion units in each segment. Mann Whitney tests (α?=?0.05) were used to determine mobility differences.

Results

The findings indicated that the AIS group had comparatively increased mobility in the periapical regions of the spine. The AIS group had an increase of 1.2° in the mid thoracic region (p?=?0.01) during flexion, an increase of 1.0° in the mid thoracic region (p?=?0.01), 1.5° in the thoracolumbar region (p?=?0.02), and 0.7° in thoracic region (p?=?0.04) during left anterior-lateral flexion, an increase of 6.0° in the upper lumbar region (p?=?0.02) during right anterior-lateral flexion, and an increase of 2.2° in the upper lumbar region during left lateral bending (p?<?0.01).

Conclusions

Participants with AIS did not have reduced mobility in sagittal or coronal motion. Contrarily, the AIS group often had a greater mobility, especially in segments directly above and below the apex. This indicates the scoliotic spine is flexible and may compensate near the apex.

     

Effect of high hydrostatic pressure treatment on isoquercetin production from rutin by commercial α-<Emphasis Type="SmallCaps">l</Emphasis>-rhamnosidase

Objectives

To optimize conversion of rutin to isoquercetin by commercial α-l-rhamnosidase using high hydrostatic pressure (HHP).

Results

The de-rhamnosylation activity of α-l-rhamnosidase for isoquercetin production was maximal at pH 6.0 and 50 °C using HHP (150 MPa). The enzyme showed high specificity for rutin. The specific activity for rutin at HHP was 1.5-fold higher than that at atmospheric pressure. The enzyme completely hydrolysed 20 mM rutin in tartary buckwheat extract after 2 h at HHP, with a productivity of 10 mM h^?1. The productivity and conversion were 2.2- and 1.5-fold higher at HHP than at atmospheric pressure, respectively.

Conclusions

This is the first report concerning the enzymatic hydrolysis of isoquercetin in tartary buckwheat at HHP.

     

Small temperature differences can improve the performance of mesophilic sludge-based digesters

Objective

To assess the effect of small temperature increases in mesophilic sludge-based digesters in order to develop and evaluate strategies for improving the biogas production in full-scale digesters.

Results

Methane production was strongly affected by small temperature differences, and this result was consistent across samples from 15 full-scale digesters. The specific methane yield varied between 42 and 97.5 ml g VS^?1 after 15 days of incubation at 35 °C, and improved when increasing the digester temperature to 39 °C. Only a limited quantity of additional gas was required to balance out the cost of heating and a positive energy balance was obtained. Further increases in temperature, in some cases, negatively affected the production when operated at 42 °C compared to 39 °C.

Conclusions

Small temperature increases should be applied to mesophilic sludge-based digesters to optimize the biogas production and is applicable to digesters operated in the lower mesophilic temperature range.

     

Production of δ-decalactone from linoleic acid via 13-hydroxy-9(Z)-octadecenoic acid intermediate by one-pot reaction using linoleate 13-hydratase and whole <Emphasis Type="Italic">Yarrowia lipolytica</Emphasis> cells

Objective

To produce δ-decalactone from linoleic acid by one-pot reaction using linoleate 13-hydratase with supplementation with whole Yarrowia lipolytica cells.

Results

Whole Y. lipolytica cells at 25 g l^?1 produced1.9 g l^?1 δ-decalactone from 7.5 g 13-hydroxy-9(Z)-octadecenoic acid l^?1 at pH 7.5 and 30 °C for 21 h. Linoleate 13-hydratase from Lactobacillus acidophilus at 3.5 g l^?1 with supplementation with 25 g Y. lipolytica cells l^?1 in one pot at 3 h produced 1.9 g l^?1 δ-decalactone from 10 g linoleic acid l^?1 via 13-hydroxy-9(Z)-octadecenoic acid intermediate at pH 7.5 and 30°C after 18 h, with a molar conversion yield of 31 % and productivity of 106 mg l^?1 h^?1.

Conclusion

To the best of our knowledge, this is the first production of δ-decalactone using unsaturated fatty acid.

     

Quinine sulfate and bacterial invasion

Background

As many patients who receive antimalarial drugs for treatment of noninfectious, inflammatory diseases are also immunosuppressed and might have a concomitant bacterial infection, we studied the effectiveness of these drugs against bacterial infections, to find out whether they could protect against (and even treat) such conditions and obviate the need for an additional antibiotic drug.

Methods

Effect of QS on bacterial growth: Escherichia coli (E. coli) HB101 pRI203 were cultured overnight at 37°C in TSB and inoculated (approx 1 × 10⁷ cells /ml) in MEM in the presence of QS at various concentrations (0, 50 and 100 μM).The effect of QS at concentration of 50 and 100 μM on the entry process of E. coli HB101 pRI203 into HeLa cells was studied under different experimental conditions: 1. QS was incubated with 3 × 10⁵ HeLa cells for 60 min at 37°C prior to infection. 2. QS was added to HeLa cell monolayers during the infection period.

Results

QS showed no antibacterial activity after 24 h of incubation.The invasive efficiency of the bacteria was significantly inhibited at a dose-dependent manner, when QS was added to HeLa cells for 60 min at 37°C prior to infection (condition 1), and to a lesser extent when added during the period of infection (condition 2).

Conclusions

Although the antimalarials are generally regarded as being inactive against most extracellular bacterial species, our results indicate that QS significantly inhibited the internalization/invasion efficacy of E. coli in the host cells.

     

Optimization of fecal sample preparation for untargeted LC-HRMS based metabolomics

Introduction

Collecting feces is easy. It offers direct outcome to endogenous and microbial metabolites.

Objectives

In a context of lack of consensus about fecal sample preparation, especially in animal species, we developed a robust protocol allowing untargeted LC-HRMS fingerprinting.

Methods

The conditions of extraction (quantity, preparation, solvents, dilutions) were investigated in bovine feces.

Results

A rapid and simple protocol involving feces extraction with methanol (1/3, M/V) followed by centrifugation and a step filtration (10 kDa) was developed.

Conclusion

The workflow generated repeatable and informative fingerprints for robust metabolome characterization.

     

Progression of idiopathic thoracic or thoracolumbar scoliosis and pelvic obliquity in adolescent patients with and without limb length discrepancy

Βackground

Both limb length inequality and scoliosis are associated with pelvic obliquity.

Methods

This is an observational study of adolescents with growth potential presenting for evaluation of thoracic or thoracolumbar idiopathic scoliosis at an outpatient pediatric orthopedic clinic. Patients were evaluated for limb length discrepancy (LLD) (using bilateral femoral head height difference), pelvic obliquity (using bilateral iliac crest height difference and sacral takeoff angle), and scoliotic curve (using Cobb angle and rotation) on full spine standing radiographs. The same radiographic parameters were measured at a follow-up visit at least 2 years later.

Results

Seventy-three consecutive patients with a mean (SD) age of 13.3 (0.2) years at initial examination were included in the study. Scoliosis (major curve Cobb angle ≥?10°) was confirmed in all 73 patients, pelvic obliquity (iliac crest height difference >?1 cm or sacral takeoff angle >?5°) appeared in 23 (31.5%) patients with scoliosis, and LLD (>?1 cm femoral head height difference) was identified in 6 (8.2%) patients with scoliosis and pelvic obliquity. At a subsequent visit, a mean of 2.8 (range 2–5.8) years later, no significant change (p?>?0.05) in limb length inequality was observed but a statistically significant increase (p?<?0.05) in scoliotic and pelvic deformity parameters was found.

Conclusions

In adolescent patient population with thoracic or thoracolumbar scoliosis, the anisomelia remains stable with growth but both the scoliotic deformity and pelvic obliquity progress.

Trial registration

MGH no 2012-P-000774/1

     

Engineering the intracellular metabolism of <Emphasis Type="Italic">Escherichia coli</Emphasis> to produce gamma-aminobutyric acid by co-localization of GABA shunt enzymes

Objectives

To direct the carbon flux from Krebs cycle into the gamma-aminobutyric acid (GABA) shunt pathway for the production of GABA by protein scaffold introduction in Escherichia coli.

Results

Escherichia coli was engineered to produce GABA from glucose by the co-localization of enzymes succinate semialdehyde dehydrogenase (GadD), GABA aminotransferase (PuuE) and GABA transporter (GadC) by protein scaffold. 0.7 g GABA l^?1 was produced from 10 g glucose l^?1 while no GABA was produced in wild type E. coli. pH 6 and 30 °C were optimum for GABA production, and GABA concentration increased to 1.12 g GABA l^?1 when 20 g glucose l^?1 was used. When competing metabolic networks were inactivated, GABA increased by 24 % (0.87 g GABA l^?1).

Conclusions

The novel GABA production system was constructed by co-localization of GABA shunt enzymes.

     

HLA-matched sibling transplantation with G-CSF mobilized PBSCs and BM decreases GVHD in adult patients with severe aplastic anemia

Background

Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is an effective treatment for severe aplastic anemia (SAA). However, graft failure and graft-versus-host disease (GVHD) are major causes of the early morbidity in Allo-HSCT.

Methods

To reduce graft failure and GVHD, we treated fifteen patients with SAA using high- dose of HSCT with both G-CSF mobilized PB and BMSCs from HLA-identical siblings to treat patients with SAA.

Results

All patients had successful bone marrow engraftment. Only one patient had late rejection. Median time to ANC greater than 0.5 × 10⁹/L and platelet counts greater than 20 × 10⁹/L was 12 and 16.5 days, respectively. No acute GVHD was observed. The incidence of chronic GVHD was 6.67%. The total three-year probability of disease-free survival was 79.8%.

Conclusion

HSCT with both G-CSF mobilized PB and BMSCs is a promising approach for heavily transfused and/or allo-immunized patients with SAA.

     

Application of two-parameter scoliometer values for predicting scoliotic Cobb angle

Background

Adolescent idiopathic scoliosis, in which obvious curves are visible in radiographic images, is also seen in combination with lumps in the back. These lumps contribute to inclination, which can be measured by a scoliometer. To the authors’ knowledge, there are no previous formulas combining thoracic and lumbar scoliometer values simultaneously to predict thoracic and lumbar Cobb angles, respectively. This study aimed to create more accurate two-parameter mathematical formulas for predicting thoracic and lumbar Cobb angles.

Methods

Between Dec. 2012 and Jan. 2013, patients diagnosed with idiopathic scoliosis in an outpatient clinic were enrolled. The maximal trunk rotations at the thoracic and lumbar regions were recorded with a scoliometer. Right asymmetry hump was deemed positive (+), and left asymmetry hump was deemed negative (?). The Cobb angles were measured with a Picture Archiving and Communication System. Statistical analysis included Pearson’s correlation coefficient, multivariate regression and Bland–Atman analysis.

Results

One-hundred and one patients were enrolled in our study. The average thoracic curve (TC) was 23.3 ± 1.8°, while the average lumbar curve (LC) was ? 23.3 ± 1.4°. The thoracic inclination (TI) and lumbar inclination (LI) were 4.5 ± 0.7 and ? 5.9 ± 0.6, respectively. The one-parameter formula for the thoracic curve was TC = 2.0 TI + 14.3 (r = 0.813); for the lumbar curve, it was LC = 0.9 LI ? 16.9 (r = 0.409). By multivariate regression, the two-parameter formulas for the thoracic and lumbar curves were TC = 2.6 TI ? 1.4 LI (r = 0.931) and LC = ? 1.5 TI + 2.0 LI (r = 0.874), respectively. The two-parameter formulas were more accurate than the one-parameter formulas.

Conclusions

Based on the results of these two-parameter formulas for thoracic and lumbar curves, the Cobb angles can be predicted more accurately by the readings of the scoliometer. Physicians and other healthcare practitioners can thus evaluate patients with scoliosis more precisely than before with a scoliometer.

     

Chemical changes in organic matter after fungal colonization in a nitrogen fertilized and unfertilized Norway spruce forest

Background and aims

Mineralization of soil organic matter (SOM) constitutes a major carbon flux to the atmosphere. The carbon use efficiency (CUE) of the saprotrophic microorganisms mineralizing SOM is integral for soil carbon dynamics. Here we investigate how the CUE is affected by temperature, metabolic conditions, and the molecular complexity of the substrate.

Methods

We incubated O-horizon soil samples (with either ¹³C–glucose or ¹³C–cellulose) from a boreal coniferous forest at 4, 9, 14, and 19 °C, and calculated CUEs based on the amount of ¹³C–CO₂ and ¹³C–labelled microbial biomass produced. The effects of substrate, temperature, and metabolic conditions (representing unlimited substrate supply and substrate limitation) on CUE were evaluated.

Results

CUE from metabolizing glucose was higher as compared to cellulose. A slight decrease in CUE with increasing temperature was observed in glucose amended samples (but only in the range 9–19 °C), but not in cellulose amended samples. CUE differed significantly with metabolic conditions, i.e. CUE was higher during unlimited growth conditions as compared to conditions with substrate limitation.

Conclusions

We conclude that it is integral to account for both differences in CUE during different metabolic phases, as well as complexity of substrate, when interpreting temperature dependence on CUE in incubation studies.

     

Construction of a highly thermostable 1,3-1,4-β-glucanase by combinational mutagenesis and its potential application in the brewing industry

Objectives

To improve the thermostability and catalytic property of a mesophilic 1,3-1,4-β-glucanase by combinational mutagenesis and to test its effect in congress mashing.

Results

A mutant β-glucanase (rE-BglTO) constructed by combinational mutagenesis showed a 25 °C increase in optimal temperature (to 70 °C) a 19.5 °C rise in T ₅₀ value and a 15.6 °C increase in melting temperature compared to wild-type enzyme. Its half-life values at 60 and 70 °C were 152 and 99 min, which were 370 and 800 % higher than those of wild-type enzyme. Besides, its specific activity and k _cat value were 42,734 U mg^?1 and 189 s^?1 while its stability under acidic conditions was also improved. In flask fermentation, the catalytic activity of rE-BglTO reached 2381 U ml^?1, which was 63 % higher than that of wild-type enzyme. The addition of rE-BglTO in congress mashing decreased the filtration time and viscosity by 21.3 and 9.6 %, respectively.

Conclusions

The mutant β-glucanase showed high catalytic activity and thermostability which indicated that rE-BglTO is a good candidate for application in the brewing industry.

     

Enhancement of the thermal and alkaline pH stability of <Emphasis Type="Italic">Escherichia coli</Emphasis> lysine decarboxylase for efficient cadaverine production

Objective

To enhance the thermal and alkaline pH stability of the lysine decarboxylase from Escherichia coli (CadA) by engineering the decameric interface and explore its potential for industrial applications.

Results

The mutant T88S was designed for improved structural stability by computational analysis. The optimal pH and temperature of T88S were 7.0 and 55 °C (5.5 and 50 °C for wild-type). T88S showed higher thermostability with a 2.9-fold increase in the half-life at 70 °C (from 11 to 32 min) and increased melting temperature (from 76 to 78 °C). Additionally, the specific activity and pH stability (residual activity after 10 h incubation) of T88S at pH 8.0 were increased to 164 U/mg and 78% (58 U/mg and 57% for wild-type). The productivity of cadaverine with T88S (284 g l-lysine L^?1 and 5 g DCW L^?1) was 40 g L^?1 h^?1, in contrast to 28 g L^?1 h^?1 with wild-type.

Conclusion

The mutant T88S showed high thermostability, pH stability, and activity at alkaline pH, indicating that this mutant is a promising biocatalyst for industrial production of cadaverine.

     

Stability of targeted metabolite profiles of urine samples under different storage conditions

Introduction

Few studies have investigated the influence of storage conditions on urine samples and none of them used targeted mass spectrometry (MS).

Objectives

We investigated the stability of metabolite profiles in urine samples under different storage conditions using targeted metabolomics.

Methods

Pooled, fasting urine samples were collected and stored at ?80 °C (biobank standard), ?20 °C (freezer), 4 °C (fridge), ~9 °C (cool pack), and ~20 °C (room temperature) for 0, 2, 8 and 24 h. Metabolite concentrations were quantified with MS using the AbsoluteIDQ? p150 assay. We used the Welch-Satterthwaite-test to compare the concentrations of each metabolite. Mixed effects linear regression was used to assess the influence of the interaction of storage time and temperature.

Results

The concentrations of 63 investigated metabolites were stable at ?20 and 4 °C for up to 24 h when compared to samples immediately stored at ?80 °C. When stored at ~9 °C for 24 h, few amino acids (Arg, Val and Leu/Ile) significantly decreased by 40% in concentration (P < 7.9E?04); for an additional three metabolites (Ser, Met, Hexose H1) when stored at ~20 °C reduced up to 60% in concentrations. The concentrations of four more metabolites (Glu, Phe, Pro, and Thr) were found to be significantly influenced when considering the interaction between exposure time and temperature.

Conclusion

Our findings indicate that 78% of quantified metabolites were stable for all examined storage conditions. Particularly, some amino acid concentrations were sensitive to changes after prolonged storage at room temperature. Shipping or storing urine samples on cool packs or at room temperature for more than 8 h and multiple numbers of freeze and thaw cycles should be avoided.

     

Does centrifugation matter? Centrifugal force and spinning time alter the plasma metabolome

Background

Centrifugation is an indispensable procedure for plasma sample preparation, but applied conditions can vary between labs.

Aim

Determine whether routinely used plasma centrifugation protocols (1500×g 10 min; 3000×g 5 min) influence non-targeted metabolomic analyses.

Methods

Nuclear magnetic resonance spectroscopy (NMR) and High Resolution Mass Spectrometry (HRMS) data were evaluated with sparse partial least squares discriminant analyses and compared with cell count measurements.

Results

Besides significant differences in platelet count, we identified substantial alterations in NMR and HRMS data related to the different centrifugation protocols.

Conclusion

Already minor differences in plasma centrifugation can significantly influence metabolomic patterns and potentially bias metabolomics studies.