Is the native ACL insertion site "completely restored" using an individualized approach to single-bundle ACL-R?

PURPOSE: The goal of individualized anatomic anterior cruciate ligament reconstruction (ACL-R) is to reproduce each patient's native insertion site as closely as possible. The amount of the native insertion site that is recreated by the tunnel aperture area is currently unknown, as are the implications of the degree of coverage. As such, the goals of this study are to determine whether individualized anatomic ACL-R techniques can maximally fill the native insertion site and to attempt to establish a crude measure to evaluate the percentage of reconstructed area as a first step towards elucidating the implications of complete footprint restoration. METHODS: This is a prospective pilot study of 45 patients who underwent primary single-bundle anatomic ACL-R from May 2011 to April 2012. Length and width of the native insertion site were measured intraoperatively. Using published guidelines, reconstruction technique and graft choice were determined to maximize the percentage of reconstructed area. Native femoral and tibial insertion site area and femoral tunnel aperture area were calculated using the formula for area of an ellipse. On the tibial side, tunnel aperture area was calculated with respect to drill diameter and drill guide angle. Percentage of reconstructed area was calculated by dividing total tunnel aperture area by the native insertion site area. RESULTS: The mean areas for the native femoral and tibial insertion sites were 83 ± 20 and 125 ± 20 mm(2), respectively. The mean tunnel aperture area for the femoral side was 65 ± 17, and 86 ± 17 mm(2) for the tibial tunnel aperture area. On average, percentage of reconstructed area was 79 ± 13 % for the femoral side, and 70 ± 12 % for the tibial side. CONCLUSION: Anatomic ACL-R does not restore the native insertion site in its entirety. Percentage of reconstructed area serves as a rudimentary tool for evaluating the degree of native insertion site coverage using current individualized anatomic techniques and provides a starting point from which to evaluate the clinical significance of complete footprint restoration. LEVEL OF EVIDENCE: IV.

Electronic screening-enhanced hole pairing in two-leg spin ladders studied by high-resolution resonant inelastic x-ray scattering at Cu M edges.

We study the electronic screening mechanisms of the effective Coulomb on-site repulsion in hole-doped Sr(14)Cu(24)O(41) compared to undoped La(6)Ca(8)Cu(24)O(41) using polarization dependent high-resolution resonant inelastic x-ray scattering at Cu M edges. By measuring the energy of the effective Coulomb on-site repulsion and the spin excitations, we estimate superexchange and hopping matrix element energies along rungs and legs, respectively. Interestingly, hole doping locally screens the Coulomb on-site repulsion reducing it by as much as 25%. We suggest that the increased ratio of the electronic kinetic to the electronic correlation energy contributes to the local superexchange mediated pairing between holes.

POEMS syndrome with Guillan-Barre syndrome-like acute onset: a case report and review of neurological progression in 30 cases.

POEMS (polyneuropathy, organomegaly, endocrinopathy, M-protein and skin changes) syndrome is a rare cause of demyelinating neuropathy with monoclonal plasma cell proliferation, and POEMS neuropathy is usually chronically progressive. Herein, the authors report a 34-year-old woman with POEMS syndrome presenting as acute polyneuropathy. Within 2 weeks of disease onset, she became unable to walk with electrodiagnostic features of demyelination and was initially diagnosed as having Guillan-Barré syndrome. Other systemic features (oedema and skin changes) developed later, and an elevated serum level of vascular endothelial growth factor led to the diagnosis of POEMS syndrome. She received high-dose chemotherapy with autologous peripheral blood stem cell transplantation, resulting in good recovery. The authors also reviewed patterns and speed of progression of neuropathy in the 30 patients with POEMS syndrome; 22 (73%) of them were unable to walk independently with the median period of 9.5 months from POEMS onset (range 0.5-51 months). Whereas the speed of neuropathy progression varies considerably among patients, some POEMS patients can show acute or subacute polyneuropathy. The early diagnosis and treatment could result in rapid improvement as shown in the present patient.

Neuromuscular transmission is not impaired in axonal Guillain--Barré syndrome.

BACKGROUND: Previous studies have shown that anti-GQ1b antibodies induce massive neuromuscular blocking. If anti-GM1 and -GD1a antibodies have similar effects on the neuromuscular junction (NMJ) in human limb muscles, this may explain selective motor involvement in axonal Guillain--Barré syndrome (GBS). METHODS: Axonal-stimulating single-fibre electromyography was performed in the extensor digitorum communis muscle of 23 patients with GBS, including 13 with the axonal form whose sera had a high titre of serum IgG anti-GM1 or -GD1a antibodies. RESULTS: All patients with axonal or demyelinating GBS showed normal or near-normal jitter, and no blocking. CONCLUSION: In both axonal and demyelinating GBS, neuromuscular transmission is not impaired. Our results failed to support the hypothesis that anti-GM1 or -GD1a antibody affects the NMJ. In GBS, impulse transmission is presumably impaired in the motor nerve terminal axons proximal to the NMJ.

Experimental observation of the crystallization of a paired holon state.

An excitation at 201 meV is observed in the doped-hole ladder cuprate Sr14Cu24O41, using ultraviolet resonance Raman scattering with incident light at 3.7 eV polarized along the rungs. The excitation is of charge nature, with a temperature independent excitation energy, and can be understood via an intraladder pair-breaking process. The intensity tracks closely the order parameter of the charge density wave in the ladder CDW(L), but persists above its transition temperature T(CDW(L)), indicating a strong local pairing above the T(CDW(L)). The 201 meV excitation vanishes in La6Ca8Cu24O(41+δ), and La5Ca9Cu24O41 which are samples with no holes in the ladders. Our results suggest that the doped holes in the ladder are composite bosons consisting of paired holons that order below T(CDW).

Distribution of ethenzamide and other ingredients on granule surfaces studied by Raman microspectroscopy and mapping.

Distributions of API and medical additives in granules were analyzed using Raman microspectroscopy and mapping. In order to clearly detect ingredients present at low levels, the characteristic peak for each ingredient was used for identification. Two granulation processes, tumbling granulation and high-shear granulation were selected to examine the feasibility of Raman microspectroscopy for investigating granules. Ethenzamide, lactose monohydrate, cornstarch and methylcellulose were used to make model granules. Methylcellulose was distributed homogeneously from the early stage in both granulation methods. Cornstarch and lactose showed similar distribution properties in high-shear granulation. It was presumed from this observation that similar chemical structures with high-hydrophilic groups in the two compounds determined their similar distribution properties. These results suggest that Raman microspectroscopy using the unique absorption peak of each ingredient can detect each ingredient in the individual pixel size (2 x 2 microm). This analytical method can contribute to evaluation of granular conditions and granulation processes.

Chemical mapping of tulobuterol in transdermal tapes using microscopic laser Raman spectroscopy.

Microscopic Laser Raman Spectroscopy and Mapping (MLRSM) technique was used to investigate the distribution of tulobuterol (TBR) crystals in transdermal tapes. TBR is one of suitable compounds for the transdermal pharmaceuticals because it has high permeability into skin. In case of TBR transdermal tapes, some commercial products also contain TBR crystals in order to control a release rate from a matrix. Therefore, the presence of TBR crystals in the matrix is a critical factor for quality assurance of this type of TDDS tapes. The model tapes prepared here employed two kinds of matrices, i.e., rubber or acrylic, which are generally used for transdermal pharmaceuticals. TBR crystals in the matrix were observed by MLRSM. Accurate observation of the distribution of TBR in the tapes was achieved by creating a Raman chemical map based on detecting unique TBR peak in each pixel. Moreover, differences in the growth of TBR crystals in the two kinds of matrices were detected by microscopic observation. MLRSM also enabled the detection of TBR crystals in commercial products. The present findings suggest that Raman micro-spectroscopic analysis would be very useful for verifying and/or assessing the quality of transdermal pharmaceuticals in development, as well as for manufacturing process control.

NIR spectroscopic investigation of two fluoroquinolones, levofloxacin and ofloxacin, and their tablets for qualitative identification of commercial products on the market.

A rapid and nondestructive identification method for ofloxacin (OFLX) and levofloxacin (LVFX) utilizing diffusion reflectance near-infrared (NIR) spectroscopy was developed. An obvious difference in spectral patterns between LVFX that is used for commercial tablets and LVFX HCl that can be purchased as a reagent at a low price was also observed. These quinolones are especially important for use as drugs against bio-terrorism because of their effectiveness against anthrax infection. Therefore, the possibility of a distribution of counterfeit drugs containing LVFX HCl on the market would be expected. NIR spectroscopic analysis would be applicable to on-site quality analysis that can be carried out easily and nondestructively.

Studies on the influence of uniformity of particle size of powder, tapping and sample replacement for diffusion reflectance quantitative NIR spectrometric analysis.

The extent of deviation factors and the influence of pre-processing of spectra for a quantitative application of reflectance NIR measurement against powder sample were examined. Lactose monohydrate (NGLM), a medical additive was used for this study. Ground lactose monohydrate (GLM) and NGLM were measured by NIR reflectance spectroscopy. In the wave number range from 12000 cm(-1) to 4000 cm(-1), the ratios of absorbance values (a.v.) between the wave numbers of GLM and NGLM were almost the same and no influence of intensity of absorbance through the measurement range was observed concerning heterogeneity of particle size. Absorbance values of NGLM were decreased with increasing number of tapping without a bit difference of the change of a.v. The several statistical parameters of a.v. from both samples were estimated. The relative standard deviation (RSD) and 95% confidence intervals (CIs) of a.v. on successive measurements at a fixed position in GLM and NGLM vials were almost the same. However, the RSD and 95% confidence of absorbance value of NGLM were larger than these GLM, i.e., RSD: 0.66% for NGLM, 0.42% for GLM. The 95% of CI of NGLM was ten times larger than that of GLM in five replacement positions. The two kinds of baseline corrections, the SNV and MSC processing were examined to evaluate the extent of influence against a.v. The 95% CI calculated from a.v. by the SNV pre-processing showed a wider range compared with that from no pre-processing and MSC pre-processing. These results suggest that the statistical confidence of a.v. would also change by pre-processing. It is important to consider the statistical confidence of a.v. for precise quantitative application of the reflectance NIR spectroscopy.

Modulation of vascular natriuretic peptide receptor gene expression in hypertensive and obese hyperglycemic rats.

Receptors for natriuretic peptide (NP) consist of three subtypes: NP-A, NP-B, and NP-C. Recent studies in cultured aortic cells have suggested a phenotype-related switching of the vascular NP receptor from NP-A to NP-B. To ascertain the biological significance of the phenomenon in vivo, we developed a sensitive and reproducible ribonuclease protection assay and determined each receptor messenger RNA (mRNA) level in the vascular vessels of stroke-prone spontaneously hypertensive rats, deoxycorticosterone acetate-salt hypertensive rats, and genetically hyperglycemic. Wistar fatty rats and in cultured aortic smooth muscle cells. The aortic NP-A receptor mRNA level was significantly up-regulated in both types of hypertensive rats, whereas the NP-B receptor mRNA level did not show any significant change. Both NP-A and NP-B receptor mRNA levels were significantly up-regulated in Wistar fatty rats compared with the control values. There was no significant up-regulation of NP-A receptor mRNA in the inferior vena cava of the stroke-prone spontaneously hypertensive rats. Although the NP-A receptor was always the predominant subtype in rat aortic tissue, NP-B receptor was the predominant subtype in aortic smooth muscle cells in culture. These findings suggest that up-regulation of the NP-A receptor, but not the subtype switching, is the major modulation of receptor gene expression in both hypertensive and diabetic rats.

Effects of recombinant human bone morphogenetic protein-2 on differentiation of cells isolated from human bone, muscle, and skin.

We investigated the effects of recombinant human BMP-2 (rhBMP-2) on differentiation of cells isolated from human bone, muscle, and skin. Cells isolated from bones of six patients (HBM-1 to HBM-6), muscle from five patients (HM-1 to HM-5), and skin from three patients (HF-1 to HF-3) were used. rhBMP-2 had no effects on proliferation of two HBM cells, but had a stimulatory effect on three HM cell samples. rhBMP-2 stimulated both alkaline phosphatase (ALP) activity in all HBM cells and parathyroid hormone (PTH)-dependent cAMP production in three of the four HBM cell samples, although the magnitudes of these stimulatory effects differed among the cells tested. Although none of the HBM cells examined produced detectable amounts of osteocalcin in the absence of 1,25-(OH)2vitamin D3, they synthesized measurable amounts of osteocalcin in its presence. rhBMP-2 inhibited 1,25-(OH)2vitamin D3-dependent osteocalcin production in all of the HBM cell samples. Transplantation of HBM-6 cells with rhBMP-2 using diffusion chambers into the peritoneal cavity of athymic mice induced formation of cartilage and bone in the diffusion chambers, but neither cartilage nor bone was formed in chambers transplanted without rhBMP-2. rhBMP-2 also stimulated ALP activity in all of the HM and HF cell samples examined and PTH-dependent cAMP production in three of four HM cell samples. rhBMP-2 induced no osteocalcin production in any of the HM or HF cells in the presence of 1,25-(OH)2vitamin D3. rhBMP-2 markedly inhibited myotube formation by all of the HM cell samples. Transplantation of HM-4 cells with rhBMP-2, using diffusion chambers, into athymic mice induced ALP-positive cells in the chambers, but neither cartilage nor bone was observed. These results suggest that rhBMP-2 is a potent stimulator of osteoblast differentiation and bone formation in human cells.

Diethylcarbamazine, antifilarial drug, inhibits microtubule polymerization and disrupts preformed microtubules.

The effect of diethylcarbamazine (DEC) on microtubules was studied by using microtubule protein prepared from porcine brain. DEC inhibited assembly of microtubules and disassembled preformed microtubules in vitro. When the reassembled or disassembled products were examined in the presence of DEC by electron microscopy, ribbon-microtubules were frequently observed. Subsequently, the effect of DEC on the cytoplasmic microtubules complex was studied. The cells used in our study were LLC-MK2. DEC inhibited proliferation of these cells, and cells grown in the presence of DEC were likely to separate from each other and became round in shape. Immunofluorescence microscopy revealed that the cells exposed to DEC were devoid of the delicate pattern of the cytoplasmic microtubule complex.

Doxazosin, an alpha1-adrenergic antihypertensive agent, decreases serum oxidized LDL.

Oxidized low-density lipoprotein (ox-LDL) is well known to play an important role in atherogenesis through the recruitment of monocytes into vessel walls and the deposition of cholesterol ester in the macrophages, which leads to the formation of lipid-rich plaque. It was assumed that only trace amounts of ox-LDL were present in plasma because the half-life of ox-LDL was only a few minutes. Recently, through the use of a monoclonal antibody against ox-LDL, a quantitative method to measure serum ox-LDL concentration has been developed. Metabolites of doxazosin, an alpha1-adrenergic antihypertensive agent, have been reported to inhibit oxidation of LDL in vitro. In this study, we investigated the in vivo effect of doxazosin on LDL oxidation using this new method to measure serum ox-LDL concentration. After the administration of doxazosin for 1 to 2 months, serum concentration of ox-LDL decreased significantly (P < .05). Although the reduction of ox-LDL concentration does not strictly indicate doxazosin's antiatherosclerotic effect, it may constitute one of doxazosin's additional weapons beside lowering blood pressure and serum lipid values in the prevention of atherosclerosis.

Defect in an intrahepatic degradation of apolipoprotein B in suncus: an animal model of hypobetalipoproteinemia.

We have previously shown that fatty liver is easily induced in suncus by starvation and that the plasma level of apolipoprotein B (apo B) is very low. We also found that hepatic acyl coenzyme A cholesterol acyltransferase (ACAT) activity is almost absent in the animals, resulting in decreased cholesteryl ester contents in the liver. A deficiency of cholesteryl ester in suncus liver may be one of the reasons for the defect in the assembly process of apo B-containing lipoproteins, leading to a low level of plasma apo B. Another possible explanation for the induction of fatty liver in suncus is a defect in apo B-processing in the liver. In this study, we investigated the hepatic synthetic rate and intrahepatic degradation of apo B using primary cultured hepatocytes derived from suncus and rats. In order to estimate intrahepatic degradation of apo B, we added N-acetylleucyl-leucynorleucinal to the culture medium as an inhibitor of apo B degradation. The basal synthesis of apo B in suncus hepatocytes was 50% of that in rat. Intracellular degradation of apo B was not observed in suncus hepatocytes, while it was obvious in rat hepatocytes. This evidence suggests that the lower secretion rate of apo B lipoprotein is not due to the intrahepatic degradation of apo B, but may be due to the low synthetic rate of apo B.

Simple and sensitive enzyme-linked immunosorbent assay for ivermectin.

A sensitive and reproducible enzyme-linked immunosorbent assay (ELISA) for the determination of the concentration of ivermectin (IVM) in biological fluids was developed. A conjugate of IVM on bovine serum albumin and poly-L-lysine was used to produce antibodies in rabbits and served as a solid-phase marker for titration of antibodies, respectively. The competitive ELISA was conducted by simultaneously incubating IVM and IVM-biotin conjugate with anti-IVM antiserum over goat anti-rabbit IgG (Fc) and then determining the amount of bound IVM-biotin with avidin-peroxidase conjugate as a tracer. The coefficient of variation for the assay was less than 10% in the range of 0.3-10 ng/ml. The limit of detection was 0.1 ng/ml. The cross-reactivities of anti-IVM antiserum with some anthelmintic drugs were negligible. Using this ELISA, serum levels of IVM were easily determined in Mongolian jirds (Meriones unguiculatus) up to 72 hr following a single oral dose of 500 microgram/kg of body weight.

Prevalence and levels of filaria-specific urinary IgG4 among children less than five years of age and the association of antibody positivity between children and their mothers.

An enzyme-linked immunosorbent assay (ELISA) to detect filaria-specific urinary IgG4 was tested in samples from 203 children less than five years old and their parents (165 mothers and 127 fathers) in Sri Lanka. There were four IgG4-positive children within 58 days after birth, suggesting the transfer of the antibody from mothers. No positive children were found between days 65 and 417. After day 1,000, the number of the positive individuals and the level of IgG4 increased quickly. The children of urinary IgG4-positive parents showed a higher IgG4 positive rate than those of negative parents. The children of positive mothers had a higher prevalence than those of negative mothers, whereas, the positivity of the fathers was not associated with that of their children. Collecting urine samples was easy to perform and well accepted because of its non-invasiveness. The ELISA will be useful for monitoring filarial infections in very young children, who are a sentinel population for evaluating the intensity of filariasis transmission and effectiveness of control measures.

Sensitive and specific enzyme-linked immunosorbent assay for the diagnosis of Wuchereria bancrofti infection in urine samples.

We developed an enzyme-linked immunosorbent assay (ELISA) that detects filaria-specific immunoglobulin G4 antibodies in unconcentrated urine. The ELISA was positive in 87 of 91 (95.6%) urine samples collected from people with Wuchereria bancrofti microfilariae, antigen, or both. Of 298 urine samples collected in Thailand, Lao People's Democratic Republic, and Japan, where no human filariasis is known, 295 (99.0%) were negative by ELISA. Various intestinal nematode and fluke infections did not interfere with the ELISA. Urine samples with sodium azide could be kept at 37 degrees C for 4 weeks, and the time of urine collection did not influence ELISA results. This ELISA can be used to identify endemic foci of filariasis.

Apolipoprotein E accelerates the efflux of cholesterol from macrophages: mechanism of xanthoma formation in apolipoprotein E deficiency.

A patient with congenitally deficient apolipoprotein (apo) E showed numerous tuberoerutive, tendon xanthomas and severe atherosclerosis, despite a low LDL concentration. In order to study the mechanism of xanthoma formation observed in apo E deficient patients, we evaluated the effect of VLDL and HDL from the patient on cholesterol ester (CE) accumulation in macrophages. The results showed that there was no difference in CE formation in macrophages among normal VLDL, the patient's VLDL and apo E containing VLDL, which was prepared by incubation of the patient's VLDL with recombinant apo E. On the other hand, apo E containing HDL, which was prepared by incubation of the patient's HDL with recombinant apo E, accelerated cholesterol efflux more effectively than did the patient's HDL and decreased intracellular CE content. Moreover, free apo E accelerated cholesterol efflux from lipid loaded macrophages. These results suggest that macrophages are prevented from transforming into foam cells by their secretion of apo E. This may also explain the marked atherosclerosis and xanthomatosis observed in the patient with apo E-deficiency.

Simple and sensitive liquid chromatography-tandem mass spectrometry method for determination of the S(+)- and R(-)-enantiomers of baclofen in human plasma and cerebrospinal fluid.

A simple and sensitive liquid chromatography-tandem mass spectrometry (LC/MS/MS) method to determine the enantiomers of the muscle relaxant baclofen in human plasma and cerebrospinal fluid (CSF) has been developed. A commercially available ultrafiltration membrane is used to prepare the sample. A chiral CROWNPAK CR(+) stationary phase column is then used to perform complete resolution of the S(+)- and R(-)-enantiomers of baclofen. This method was used to analyze human plasma and CSF spiked with baclofen, and the calibration curves for both biologic samples were linear over a concentration range of 0.15-150 ng enantiomer/ml. The lower limit of quantification was 0.15 ng enantiomer/ml in both fluids. Finally, the method was tested with an artificial CSF as an alternative to authentic human CSF. The results showed that no matrix effects and no interfering peaks were observed using this artificial CSF.

Studies of anaerobic bacteria in chronic otitis media.

In the 760 cases of chronic otitis media studied from 1970 to 1976, the detection ratio of anaerobic bacteria and the relation between the nature of anaerobic bacteria and the conditions of focus were investigated. In 8.2% of 760 cases 9 kinds and 81 strains of anaerobic bacteria was detected. Most of the anaerobic bacteria were Peptococcus sp. or Bacteroides sp. and in general non-sporogenic anaerobic bacteria accounted for 97.5%. Anaerobes were frequently detected in cases where the middle ear cavity was filled with cholesteatoma or granulation and in cases of infection recurring after operation.