Pregnancies achieved with testicular and ejaculated spermatozoa in combination with intracytoplasmic sperm injection in men with totally or initially immotile spermatozoa in the ejaculate

The efficacy of intracytoplasmic sperm injection (ICSI) employingtesticular and ejaculated spermatozoa was assessed in 24 coupleswith totally or initially immotile spermatozoa. No criteriawere employed in selecting which patients would be treated withtesticular or ejaculated spermatozoa. The men were chosen atrandom. Testicular spermatozoa obtained by testicular spermextraction were used in 14 and ejaculated spermatozoa were usedin 10 of these couples. In all cases, asthenozoospermia wastotal in their basal semen sample. In 12 male partners, spermatozoawere totally immotile before and after Percoll gradient fractionation(totally immotile). In the remaining 12 men, spermatozoa initiallyshowed a total absence of motility; however, some of the spermatozoahad showed very poor motility (0.1%) after Percoll gradientfractionation and a 13–2.0 h incubation period (initiallyimmotile). Of these 24 total asthenozoospermic males, 14 alsohad total terato-zoospermia. The fertilization and cleavagerates in the testicular and ejaculated sperm groups were 533and 963 and 543 and 94.4% respectively. One cycle resulted incomplete fertilization failure, and in 23 embryo transfer cyclesa total of 10 pregnancies were obtained (41.6%). Eight pregnancieswere achieved in the testicular sperm group, while only twopregnancies were obtained in the ejaculated sperm group. Fourpregnancies, two from the ejaculated sperm group and two fromthe testicular sperm group, resulted in clinical abortions inthe first trimester. Of the remaining six pregnancies, two havealready resulted in healthy births and four pregnancies arenow in the second or third trimester in the testicular spermgroup. Using testicular spermatozoa in combination with ICSIcan be an alternative mode of treatment in cases with totallyor initially immotile spermatozoa in the ejaculate. Very lowpregnancy rates have been obtained and no ongoing pregnancyhas been achieved using ejaculated spermatozoa in these cases.     

Subtelomeric deletions detected in patients with idiopathic mental retardation using multiplex ligation-dependent probe amplification (MLPA)

Subtelomeric rearrangements are responsible for 5% to 10% of cases of unexplained mental retardation. Despite their clinical relevance, methods to screen for these cytogenetically invisible abnormalities on a routine base are scarce. We screened patients with idiopathic mental retardation for subtelomeric aberrations using multiplex ligation-dependent probe amplification (MLPA). This recently developed technique is based on PCR amplification of ligated probes hybridized to chromosome ends. Currently, 41 telomeres can be screened in just two multiplex reactions. Four subtelomeric rearrangements (5.3%) were detected in a group of 75 patients with mild to severe mental retardation in combination with dysmorphic features and/or a familial history of mental retardation: two terminal 1p deletions, a terminal 1q deletion, and a terminal 3p deletion. Deletions could be verified by FISH and marker analysis. In one case the MLPA indicated a terminal 21q deletion due to a 3-bp deletion at the site of the probe, giving a false-positive rate of 1.3%. This study demonstrates that MLPA is a fast and reliable screening method, potentially suitable for use in routine diagnostics.     

Absorbed dose to water based dosimetry versus air kerma based dosimetry for high-energy photon beams: an experimental study

In recent years, a change has been proposed from air kerma based reference dosimetry to absorbed dose based reference dosimetry for all radiotherapy beams of ionizing radiation. In this paper, a dosimetry study is presented in which absorbed dose based dosimetry using recently developed formalisms was compared with air kerma based dosimetry using older formalisms. Three ionization chambers of each of three different types were calibrated in terms of absorbed dose to water and air kerma and sent to five hospitals. There, reference dosimetry with all the chambers was performed in a total of eight high-energy clinical photon beams. The selected chamber types were the NE2571, the PTW-30004 and the Wellh?fer-FC65G (previously Wellh?fer-IC70). Having a graphite wall, they exhibit a stable volume and the presence of an aluminium electrode ensures the robustness of these chambers. The data were analysed with the most important recommendations for clinical dosimetry: IAEA TRS-398, AAPM TG-51, IAEA TRS-277, NCS report-2 (presently recommended in Belgium) and AAPM TG-21. The necessary conversion factors were taken from those protocols, or calculated using the data in the different protocols if data for a chamber type are lacking. Polarity corrections were within 0.1% for all chambers in all beams. Recombination corrections were consistent with theoretical predictions, did not vary within a chamber type and only slightly between different chamber types. The maximum chamber-to-chamber variations of the dose obtained with the different formalisms within the same chamber type were between 0.2% and 0.6% for the NE2571, between 0.2% and 0.6% for the PTW-30004 and 0.1% and 0.3% for the Wellh?fer-FC65G for the different beams. The absorbed dose results for the NE2571 and Wellh?fer-FC65G chambers were in good agreement for all beams and all formalisms. The PTW-30004 chambers gave a small but systematically higher result compared to the result for the NE2571 chambers (on the average 0.1% for IAEA TRS-277, 0.3% for NCS report-2 and AAPM TG-21 and 0.4% for IAEA TRS-398 and AAPM TG-51). Within the air kerma based protocols, the results obtained with the TG-21 protocol were 0.4-0.8% higher mainly due to the differences in the data used. Both absorbed dose to water based formalisms resulted in consistent values within 0.3%. The change from old to new formalisms is discussed together with the traceability of calibration factors obtained at the primary absorbed dose and air kerma standards in the reference beams (60Co). For the particular situation in Belgium (calibrations at the Laboratory for Standard Dosimetry of Ghent) the change amounts to 0.1-0.6%. This is similar to the magnitude of the change determined in other countries.     

Foxp3+ regulatory T cells are activated in spite of B7‐CD28 and CD40‐CD40L blockade

Costimulatory signals are required for priming and activation of naive T cells, while it is less clear how they contribute to induction of regulatory T (Treg)‐cell activity. We previously reported that the blockade of the B7‐CD28 and CD40L‐CD40 interaction efficiently suppresses allogeneic T‐cell activation in vivo. This was characterized by an initial rise in Foxp3⁺ cells, followed by depletion of host‐reactive T cells. To further investigate effects of costimulatory blockade on Treg cells, we used an in vitro model of allogeneic CD4⁺ cell activation. When CTLA‐4Ig and anti‐CD40L mAb (MR1) were added to the cultures, T‐cell proliferation and IL‐2 production were strongly reduced. However, Foxp3⁺ cells proliferated and acquired suppressive activity. They suppressed activation of syngeneic CD4⁺ cells much more efficiently than did freshly isolated Treg cells. CD4⁺ cells activated by allogeneic cells in the presence of MR1 and CTLA‐4Ig were hyporesponsive on restimulation, but their response was restored to that of naive CD4⁺ cells when Foxp3⁺ Treg cells were removed. We conclude that natural Treg cells are less dependent on B7‐CD28 or CD40‐CD40L costimulation compared with Foxp3^? T cells. Reduced costimulation therefore alters the balance between Teff and Treg‐cell activation in favor of Treg‐cell activity.     

Fluorescent non-porous silica nanoparticles for long-term cell monitoring: Cytotoxicity and particle functionality

Inorganic nanoparticles such as silica particles offer many exciting possibilities for biomedical applications. However, the possible toxicity of these particles remains an issue of debate that seriously impedes their full exploitation. In the present work, commercially available fluorescent silica nanoparticles 25, 45 and 75 nm in diameter optimized for cell labelling (C-Spec® particles) are evaluated with regard to their effects on cultured cells using a novel multiparametric setup. The particles show clear concentration and size-dependent effects, where toxicity is caused by the number and total surface area of cell-associated particles. Cell-associated particles generate a short burst of oxidative stress that, next to inducing cell death, affects cell signalling and impedes cell functionality. For cell labelling purposes, 45 nm diameter silica particles were found to be optimally suited and no adverse effects were noticeable at concentrations of 50 μg ml^?1 or below. At this safe concentration, the particles were found to still allow fluorescence tracking of cultured cells over longer time periods. In conclusion, the data shown here provide a suitable concentration of silica particles for fluorescent cell labelling and demonstrate that at safe levels, silica particles remain perfectly suitable for fluorescent cell studies.     

Clinical Assessment of Scapular Positioning in Musicians: An Intertester Reliability Study

Context:

The reliability of the measurement of the distance between the posterior border of the acromion and the wall and the reliability of the modified lateral scapular slide test have not been studied. Overall, the reliability of the clinical tools used to assess scapular positioning has not been studied in musicians.

Objective:

To examine the intertester reliability of scapular observation and 2 clinical tests for the assessment of scapular positioning in musicians.

Design:

Intertester reliability study.

Setting:

University research laboratory.

Patients or Other Participants:

Thirty healthy student musicians at a single university.

Main Outcome Measure(s):

Two assessors performed a standardized observation protocol, the measurement of the distance between the posterior border of the acromion and the wall, and the modified lateral scapular slide test. Each assessor was blinded to the other''s findings.

Results:

The intertester reliability coefficients (κ) for the observation in relaxed position, during unloaded movement, and during loaded movement were 0.41, 0.63, and 0.36, respectively. The κ values for the observation of tilting and winging at rest were 0.48 and 0.42, respectively; during unloaded movement, the κ values were 0.52 and 0.78, respectively; and with a 1-kg load, the κ values were 0.24 and 0.50, respectively. The intraclass correlation coefficient (ICC) of the measurement of the acromial distance was 0.72 in relaxed position and 0.75 with the participant actively retracting both shoulders. The ICCs for the modified lateral scapular slide test varied between 0.63 and 0.58.

Conclusions:

Our results demonstrated that the modified lateral scapular slide test was not a reliable tool to assess scapular positioning in these participants. Our data indicated that scapular observation in the relaxed position and during unloaded abduction in the frontal plane was a reliable assessment tool. The reliability of the measurement of the distance between the posterior border of the acromion and the wall in healthy musicians was moderate.     

Fast reconstruction of 3D time-of-flight PET data by axial rebinning and transverse mashing

Faster scintillators like LaBr(3) and LSO have sparked renewed interest in PET scanners with time-of-flight (TOF) information. The TOF information adds another dimension to the data set compared to conventional three-dimensional (3D) PET with the size of the projection data being multiplied by the number of TOF bins. Here we show by simulations and analytical reconstruction that angular sampling for two-dimensional (2D) TOF PET can be reduced significantly compared to what is required for conventional 2D PET. Fully 3D TOF PET data, however, have a wide range of oblique and transverse angles. To make use of the smaller necessary angular sampling we reduce the 3D data to a set of 2D histoprojections. This is done by rebinning the 3D data to 2D data and by mashing these 2D data into a limited number of angles. Both methods are based on the most likely point given by the TOF measurement. It is shown that the axial resolution loss associated with rebinning reduces with improved timing resolution and becomes less than 1 mm for a TOF resolution below 300 ps. The amount of angular mashing that can be applied without tangential resolution loss increases with improved TOF resolution. Even quite coarse angular mashing (18 angles out of 324 measured angles for 424 ps) does not significantly reduce image quality in terms of the contrast or noise. The advantages of the proposed methods are threefold. Data storage is reduced to a limited number of 2D histoprojections with TOF information. Compared to listmode format we have the advantage of a predetermined storage space and faster reconstruction. The method does not require the normalization of projections prior to rebinning and can be applied directly to measured listmode data.     

Reconstruction of 2D PET data with Monte Carlo generated system matrix for generalized natural pixels

In discrete detector PET, natural pixels are image basis functions calculated from the response of detector pairs. By using reconstruction with natural pixel basis functions, the discretization of the object into a predefined grid can be avoided. Here, we propose to use generalized natural pixel reconstruction. Using this approach, the basis functions are not the detector sensitivity functions as in the natural pixel case but uniform parallel strips. The backprojection of the strip coefficients results in the reconstructed image. This paper proposes an easy and efficient way to generate the matrix M directly by Monte Carlo simulation. Elements of the generalized natural pixel system matrix are formed by calculating the intersection of a parallel strip with the detector sensitivity function. These generalized natural pixels are easier to use than conventional natural pixels because the final step from solution to a square pixel representation is done by simple backprojection. Due to rotational symmetry in the PET scanner, the matrix M is block circulant and only the first blockrow needs to be stored. Data were generated using a fast Monte Carlo simulator using ray tracing. The proposed method was compared to a listmode MLEM algorithm, which used ray tracing for doing forward and backprojection. Comparison of the algorithms with different phantoms showed that an improved resolution can be obtained using generalized natural pixel reconstruction with accurate system modelling. In addition, it was noted that for the same resolution a lower noise level is present in this reconstruction. A numerical observer study showed the proposed method exhibited increased performance as compared to a standard listmode EM algorithm. In another study, more realistic data were generated using the GATE Monte Carlo simulator. For these data, a more uniform contrast recovery and a better contrast-to-noise performance were observed. It was observed that major improvements in contrast recovery were obtained with MLEM when the correct system matrix was used instead of simple ray tracing. The correct modelling was the major cause of improved contrast for the same background noise. Less important factors were the choice of the algorithm (MLEM performed better than ART) and the basis functions (generalized natural pixels gave better results than pixels).     

The use of human tissue in epidemiological research; ethical and legal considerations in two biobanks in Belgium

This paper discusses the legal implications of setting up two new biobanks in Belgium. The first is hospital-based and will archive tissue from patients with haematologic cancer, whereas the second is linked to a general practice based morbidity registry and will involve storage of blood samples. To date, Belgium has no specific legislation that regulates storage of human tissue and related databases. Several issues concerning the protection of individuals with regard to the processing of personal medical data are discussed from the existing privacy legislation. We will address the principle of consent (broad versus specific) and the type of data recorded (anonymous, encoded and identifiable) for both biobanks.     

The outcome of proximal fifth metatarsal fractures: redefining treatment strategies

Background

To optimize the treatment strategy and reduce treatment costs of proximal fifth metatarsal fractures, clinical and patient-reported outcome, and its determinants were addressed.

Methods

A retrospective adult cohort study including 152 proximal fifth metatarsal fractures: 121 nonoperatively and 31 operatively treated. In the operative group, 21 were zone 1 and 10 zone 2 fractures. Median follow-up was 37.5 (IQR 20.8–52.3) months with a minimal follow-up of 6 months. Twenty-three demographic, fracture, and treatment characteristics were assessed as well as the healthcare costs. Outcome was assessed using the patient files, anterior-posterior and oblique X-rays, foot function index (FFI), visual analog score (VAS), and SF-36 questionnaires.

Results

The median FFI, physical SF-36, and VAS scores did not significantly differ between nonoperatively and operatively treated patients. The FFI and physical SF-36 were predominantly affected by a history of mobility impairment and pre-existent cardiovascular diseases, whereas mental SF-36 correlated significantly with higher ASA-score. Overall complication rate was 5.9% (4.1 vs. 12.9%; p?=?0.065, nonoperative vs. operative, respectively). Nonunion was recorded in only one (nonoperatively) treated patient. The total healthcare costs for operative treatment were 4.2 times higher compared to nonoperative treatment (€1960 vs. €463 per patient, respectively).

Conclusion

Overall, the clinical and patient-reported outcome was good. The foot function and quality of life were mainly affected by comorbidity, rather than fracture and treatment-related variables. Although nonoperatively treated patients indicated decreased mental quality of life, our study indicates that proximal fifth metatarsal fractures can safely be treated nonoperatively without the risk of nonunion, with fewer complications and lower healthcare costs.

Level of evidence

3.