Search for mutations involved in human globozoospermia

BACKGROUND: Globozoospermia is a severe form of teratozoospermia characterized by round-headed sperm with an absence of acrosomes. Family cases of globozoopermia suggest that this pathology has genetic origins, but the mode of inheritance remains unknown. So far, no responsible genes have been identified. Recently, a mouse lacking the casein kinase IIalpha' (encoded by the Csnk2a2 gene) was described. This mutant mouse presents a single phenotype reminiscent of that seen in human globozoospermia. Interestingly, the fission yeast orthologue (orb5) exhibits, when mutated, a spherical phenotype. Casein kinase II is a heterotetramer, composed of two catalytic subunits alpha or alpha' and two regulatory beta subunits (encoded by the Csnk2b gene). METHODS and RESULTS: Based on the evolution conservation, phenotypes observed in mouse and yeast mutant and the structure of casein kinase II, we analysed Csnk2a2 and Csnk2b genes in six patients with globozoospermia and 10 fertile controls. Genomic DNA was extracted from peripheral blood and PCR was performed to amplify Csnk2a2 and Csnk2b genes before sequencing. CONCLUSION: No mutation was identified among these six patients. Further work is needed, with a larger patient data set, to identify putative genes involved in this form of male infertility.     

Optimization of Processing Steps for Superior Functional Properties of (Ba,Ca)(Zr,Ti)O3 Ceramics

Lead-free piezoelectric ceramics with nominal composition at morphotropic phase boundary Ba_0.85Ca_0.15Ti_0.9Zr_0.1O₃ (BCTZ) prepared by different processing routes and sintered either by conventional solid-state reaction or by spark plasma sintering (SPS) techniques were comparatively investigated to observe the role of structural modifications and of microstructures on the dielectric, ferroelectric, piezoelectric and electrocaloric responses. The ceramics presented relative densities from 75% to 97% and showed variations in their phase composition as a result of variable mixing and different synthesis and sintering parameters providing local compositional heterogeneity. As result, all of the ceramics showed diffuse phase transition and ferroelectric switching responses, with parameters affected mostly by density (P_r between 3.6 to 10.1 μC/cm²). High values for the electrocaloric response in the Curie range were found for the ceramics with predominantly orthorhombic character. Field-induced structural modifications were probed by tunability anomalies and by XRD experiments in remanence conditions. Piezoelectric effects with notably high figure of merit values were assigned to the better densification and poling efficiency of BCTZ ceramics.     

Case report: birth after preimplantation genetic diagnosis of a subtle mutation in SMN1 gene

Spinal muscular atrophy (SMA) preimplantation genetic diagnosis (PGD) has been available since 1998. Protocols are based on the detection of the homozygous deletion of exon 7, which are present in 90-98% of SMA patients. A couple where the woman was a heterozygous carrier of the usual SMN1 Del7 mutation and the man was a heterozygous carrier of pMet263Arg substitution in exon 6 of SMN1 gene was referred for PGD. The usual PGD test being unsuitable for this couple, we developed a novel duplex polymerase chain reaction (PCR)-based PGD test for the detection of the mutation pMet263Arg by allele specific amplification, combined with the amplification of D5S641 extragenic polymorphic marker. PCR conditions were established using single control lymphoblasts and lymphocytes from the pMet263Arg substitution carrier. Amplification was obtained in 100% of the 86 single cells tested, amplification refractory mutation system (ARMS) PCR was specific in 100% of single cells tested and a complete genotype (mutation plus D5S641) was achieved in 88% of them. A PGD cycle was performed successfully and a pregnancy was obtained. An unaffected girl was born and postnatal diagnosis confirmed PGD results. This is the first PGD described for SMA because of another mutation than the major homozygous exon 7 deletion of SMN1. In the future, a similar strategy could be adopted for other subtle mutations of this gene.     

Volumetric-modulated arc stereotactic body radiotherapy for prostate cancer: dosimetric impact of an increased near-maximum target dose and of a rectal spacer

Objective:

In volumetric-modulated arc therapy (VMAT) prostate stereotactic body radiotherapy (SBRT), dose coverage of the planning target volume (PTV) becomes challenging when the sparing of rectum, bladder and urethra is strictly pursued. Our current 35-Gy-in-five-fraction plans only assure 33.2 Gy to ≥95% PTV (V33.2PTV ≥ 95%). Looking for an improved V33.2PTV, increased near-maximum target dose (D_2%) and prostate–rectum spacer insertion were tested.

Methods:

For 11 patients, two VMAT plans, with D_2% ≤ 37.5 Gy (Hom) or D_2% ≤ 40.2 Gy (Het), on each of two CT studies, before or after spacer insertion, were computed. All plans assured V33.2PTV ≥95%, and <1 cm³ of rectum, bladder and urethra receiving ≥35 Gy. By hypothesis testing, several dose–volume metrics for target coverage and rectal sparing were compared across the four groups of plans. The impact of spacer insertion on the fractions of rectum receiving more than 18, 28 and 32 Gy (VXr) was further tested by linear correlation analysis.

Results:

By hypothesis testing, the increased D_2% was associated with improvements in target coverage, whereas spacer insertion was associated with improvements in both target coverage and rectal VXr. By linear correlation analysis, spacer insertion was related to the reductions in rectal VXr for X ≥ 28 Gy.

Conclusion:

A slightly increased D_2% or the use of spacer insertion was each able to improve V33.2PTV. Their combined use assured V33.2PTV ≥ 98% to all our patients. Spacer insertion was further causative for improvements in rectal sparing.

Advances in knowledge:

For VMAT plans in prostate SBRT, the distinct dosimetric usefulness of increased D_2% and of the use of spacer insertion were validated in terms of target coverage and rectal sparing.     

Mitral Valve Surgery in 6 Patients after Failed MitraClip Therapy

The MitraClip percutaneous mitral valve repair system, developed as an option for percutaneous mitral repair, was clinically introduced in 2007. From 2010 through 2012, 6 of our patients underwent mitral valve surgery after MitraClip failure. Their mean age was 75 ± 7.7 years (range, 62–87 yr). Three had undergone cardiac surgery previously. In 5 of the 6 patients, mitral regurgitation recurred after initially successful MitraClip deployment and was the indication for surgery. The mean interval between MitraClip implantation and surgery was 106 ± 86 days (range, 0–238 d).Mitral valve repair was feasible in 3 patients; the others underwent valve replacement. All the patients underwent additional cardiac procedures, because the MitraClip worsened existing conditions. Echocardiograms revealed sufficient valvular repairs. Two patients died during hospitalization, one of cerebral infarction and the other of bowel ischemia.Mitral valve repair after failed MitraClip therapy can be complex and a surgical challenge. Careful consideration should be given to appropriate patient selection for MitraClip therapy, because the MitraClip can cause existing pathologic valvular conditions to deteriorate substantially. The interval between MitraClip failure and corrective surgery should be as short as possible. The primary indication is an issue of ongoing discussion.     

Evaluation of monoenergetic late iodine enhancement dual-energy computed tomography for imaging of chronic myocardial infarction

Objectives

To evaluate image quality and diagnostic accuracy of selective monoenergetic reconstructions of late iodine enhancement (LIE) dual-energy computed tomography (DECT) for imaging of chronic myocardial infarction (CMI).

Methods

Twenty patients with a history of coronary bypass surgery underwent cardiac LIE-DECT and late gadolinium enhancement (LGE) magnetic resonance imaging (MRI). LIE-DECT images were reconstructed as selective monoenergetic spectral images with photon energies of 40, 60, 80, and 100 keV and the standard linear blending setting (M_0.6). Images were assessed for late enhancement, transmural extent, signal characteristics and subjective image quality.

Results

Seventy-nine myocardial segments (23 %) showed LGE. LIE-DECT detected 76 lesions. Images obtained at 80 keV and M_0.6 showed a high signal-to-noise ratio (15.9; 15.1), contrast-to-noise ratio (4.2; 4.0) and sensitivity (94.9 %; 92.4 %) while specificity was identical (99.6 %). Differences between these series were not statistically significant. Transmural extent of LIE was overestimated in both series (80 keV: 40 %; M_0.6: 35 %) in comparison to MRI. However, observers preferred 80 keV in 13/20 cases (65 %, κ?=?0.634) over M_0.6 (4/20 cases) regarding subjective image quality.

Conclusions

Post-processing of LIE-DECT data with selective monoenergetic reconstructions at 80 keV significantly improves subjective image quality while objective image quality shows no significant difference compared to standard linear blending.

Key Points

? Late enhancement dual-energy CT allows for detection of chronic myocardial infarction. ? Monoenergetic reconstructions at 80 keV significantly improve subjective image quality. ? 80 keV and standard linear blending reconstructions show no significant differences. ? Extent of CMI detected with LIE-DECT is overestimated compared with MRI.     

A blood‐based biomarker panel for stratifying current risk for colorectal cancer

Colorectal cancer (CRC) is often curable and preventable using current screening modalities. Unfortunately, screening compliance remains low, partly due to patient dissatisfaction with faecal/endoscopic testing. Recent guidelines advise CRC screening should begin with risk stratification. A blood‐based test providing clinically actionable CRC risk information would likely improve screening compliance and enhance clinical decision making. We analyzed 196 gene expression profiles to select candidate CRC biomarkers. qRT‐PCR was performed on 642 samples to develop a 7‐gene biomarker panel using 112 CRC/120 controls (training set) and 202 CRC/208 controls (independent, blind test set). Panel performance characteristics and disease prevalence (0.7%) were then used to develop a scale assessing an individual's current risk of having CRC based on his/her gene signature. A 7‐gene panel (ANXA3, CLEC4D, LMNB1, PRRG4, TNFAIP6, VNN1 and IL2RB) discriminated CRC in the training set (area under the receiver‐operating‐characteristic curve (ROC AUC), 0.80; accuracy, 73%; sensitivity, 82%; specificity 64%). The independent blind test set confirmed performance (ROC AUC, 0.80; accuracy, 71%; sensitivity, 72%; specificity, 70%). Individual gene profiles were compared against the population results and used to calculate the current relative risk for CRC. We have developed a 7‐gene, blood‐based biomarker panel that can stratify subjects according to their current relative risk across a broad range in an average‐risk population. Across the continuous spectrum of risk as defined by the current relative risk scale, it is possible to identify clinically meaningful reference points that can assist patients and physicians in CRC screening decision making.     

Catalytic therapy of cancer with ascorbate and extracts of medicinal herbs

Catalytic therapy (CT) is a cancer treatment modality based on the generation of reactive oxygen species (ROS) using a combination of substrate molecules and a catalyst. The most frequently used substrate/catalyst pair is ascorbate/Co phthalocyanine (PcCo). In the present work, herb extracts containing pigments have been studied as a catalyst in place of PcCo. Extracts from herbs are expected to have efficiency comparable with that of phthalocyanines but as natural products, to exhibit fewer side effects. The present studies demonstrate that a combined use of ascorbate and herbal extracts results in ROS production and a significant decrease in the number of cancer cells after a single in vitro treatment. Treatment with ascorbate in conjunction with extracts prepared from several medicinal herbs stimulated apoptosis and disrupted the cell cycle. The number of cells accumulating in the sub-G0/G1 stage of the cell cycle was increased 2- to 7-fold, and cells in G(2)/M increased 1.5- to 20-fold, indicating that the treatment protocol was highly effective in suppressing DNA synthesis and potentially reflecting DNA damage in the tumor cells. In addition, 20-40% of the cells underwent apoptosis within 24 h of completing treatment. Our results suggest that herbal extracts can function as CT catalysts in the treatment of cancer.