Specific haplotypes of the CALPAIN-5 gene are associated with polycystic ovary syndrome

BACKGROUND: Polycystic ovary syndrome (PCOS) is a common endocrine disorder in women of reproductive age. The aim of the present study was to investigate the role of CALPAIN-5 (CAPN5) gene in PCOS susceptibility. METHODS: We analysed four intronic polymorphisms of the CAPN5 gene in 148 well-characterized women with PCOS and 606 unrelated controls. We performed a case-control study and an intracohort analysis of clinical characteristics associated with PCOS. RESULTS: Analysis of haplotypes distribution between PCOS population compared to controls showed a strong deviation (P = 0.00029). The haplotypes GGCA and GGTG were overrepresented in PCOS patients (P = 0.009 and P = 0.001, respectively). In addition, we identified several CAPN5 haplotypes associated with phenotypic differences observed between PCOS patients, such as the presence of obesity (P = 0.02), cardiovascular complications (P = 0.02), familial antecedents of obesity (P = 0.003) and of hypertension (P = 0.007) and type 2 diabetes mellitus aggregation (P = 0.04). CONCLUSIONS: These results suggest a role of CAPN5 gene in PCOS susceptibility in humans. Moreover, novel candidate risk alleles have been identified, within CAPN5 gene, which could be associated with important phenotypic and prognosis differences observed in PCOS patients.     

Genetic relatedness and antimicrobial resistance determinants among clinical isolates of enterococci from Cuba

This study describes the genetic relationships and antimicrobial resistance determinants found among 99 clinical isolates of enterococci from 15 different hospitals in Cuba. Pulsed-field gel electrophoresis SmaI analysis demonstrated a high degree of genetic diversity. A limited number of multiresistant Enterococcus faecalis clones, showing resistance to three or more families of antimicrobial agents, were detected simultaneously in different institutions, suggesting inter-hospital circulation of selected clones, and/or selection of particular clones following their introduction into the hospital environment. Antimicrobial resistance determinants, including erm(B), aac(6')-aph(2'), aph(3'), ant(6), vanB (E. faecalis) and vanA (Enterococcus faecium) were detected by PCR in various isolates.     

The effect of pronuclear morphology on early development and chromosomal abnormalities in cleavage-stage embryos

BACKGROUND: Pronuclear (PN) zygote morphology has been proposed as a useful tool for selecting the best embryos for transfer. METHODS: PN morphology was recorded in 888 zygotes and classified according to similar/different PN size [groups A (n = 816) and B (n = 72)] and to the number, distribution and synchrony of nucleolar precursor bodies (NPB): subgroup I, pronuclei with 3-4 polarized NPB; subgroup II, 5-7 synchronic polarized NPB or 7-10 NPB distributed randomly; and subgroup III, morphologies other than those of groups I or II. Embryo development and chromosomal abnormalities were evaluated for each PN pattern. RESULTS: In patients aged 37 years, statistical differences among groups were not observed. CONCLUSIONS: In patients aged 37 years, this correlation does not exist.     

Whole-blastocyst culture followed by laser drilling technology enhances the efficiency of inner cell mass isolation and embryonic stem cell derivation from good- and poor-quality mouse embryos: new insights for derivation of human embryonic stem cell lines

The optimization of human embryonic stem (hES) cell line derivation methods is challenging because many worldwide laboratories have neither access to spare human embryos nor ethical approval for using supernumerary human embryos for hES cell derivation purposes. Additionally, studies performed directly on human embryos imply a waste of precious human biological material. In this study, we developed a new strategy based on the combination of whole-blastocyst culture followed by laser drilling destruction of the trophoectoderm for improving the efficiency of inner cell mass (ICM) isolation and ES cell derivation using murine embryos. Embryos were divided into good- and poor-quality embryos. We demonstrate that the efficiency of both ICM isolation and ES cell derivation using this strategy is significantly superior to whole-blastocyst culture or laser drilling technology itself. Regardless of the ICM isolation method, the ES cell establishment depends on a feeder cell growth surface. Importantly, this combined methodology can be successfully applied to poor-quality blastocysts that otherwise would not be suitable for laser drilling itself nor immunosurgery in an attempt to derive ES cell lines due to the inability to distinguish the ICM. The ES cell lines derived by this combined method were characterized and shown to maintain a typical morphology, undifferentiated phenotype, and in vitro and in vivo three germ layer differentiation potential. Finally, all ES cell lines established using either technology acquired an aneuploid karyotype after extended culture periods, suggesting that the method used for ES cell derivation does not seem to influence the karyotype of the ES cells after extended culture. This methodology may open up new avenues for further improvements for the derivation of hES cells, the majority of which are derived from frozen, poor-quality human embryos.     

腹针治疗周围性面瘫选穴规律研究

目的 研究腹针治疗周围性面瘫的选穴规律。方法 通过计算机及手工检索,选取近15年有关腹针治疗周围性面瘫的相关文献,以临床研究、疗效观察、现代对比实验研究为依据,进行分析归纳,探讨腹针的选穴规律。结果 腹针治疗周围性面瘫主要取任脉、肾经、胃经及脾经上的穴位及腹部新穴,使用频率依次为55.70%、15.44%、14.09%、2.68%、12.08%;腹针治疗周围性面瘫选用的标志性穴位组合规律:主要有引气归元、腹四关、天地针、调脾气,其使用频率依次为48.48%、27.27%、12.12%、12.12%。结论 合理运用腹针,遵循其选穴规律,可有效治疗周围性面瘫。     

A masked least-squares smoothing procedure for artifact reduction in scanning-EMG recordings

Scanning-EMG is an electrophysiological technique in which the electrical activity of the motor unit is recorded at multiple points along a corridor crossing the motor unit territory. Correct analysis of the scanning-EMG signal requires prior elimination of interference from nearby motor units. Although the traditional processing based on the median filtering is effective in removing such interference, it distorts the physiological waveform of the scanning-EMG signal. In this study, we describe a new scanning-EMG signal processing algorithm that preserves the physiological signal waveform while effectively removing interference from other motor units. To obtain a cleaned-up version of the scanning signal, the masked least-squares smoothing (MLSS) algorithm recalculates and replaces each sample value of the signal using a least-squares smoothing in the spatial dimension, taking into account the information of only those samples that are not contaminated with activity of other motor units. The performance of the new algorithm with simulated scanning-EMG signals is studied and compared with the performance of the median algorithm and tested with real scanning signals. Results show that the MLSS algorithm distorts the waveform of the scanning-EMG signal much less than the median algorithm (approximately 3.5 dB gain), being at the same time very effective at removing interference components.

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Graphical Abstract The raw scanning-EMG signal (left figure) is processed by the MLSS algorithm in order to remove the artifact interference. Firstly, artifacts are detected from the raw signal, obtaining a validity mask (central figure) that determines the samples that have been contaminated by artifacts. Secondly, a least-squares smoothing procedure in the spatial dimension is applied to the raw signal using the not contaminated samples according to the validity mask. The resulting MLSS-processed scanning-EMG signal (right figure) is clean of artifact interference.

     

HLA-DRB1 associations with disease susceptibility and clinical course in Australians with multiple sclerosis

Human leucocyte antigen (HLA)-DRB1*1501 and other class II alleles influence susceptibility to multiple sclerosis (MS), but their contribution if any to the clinical course of MS remains uncertain. Here, we have investigated DRB1 alleles in a large sample of 1230 Australian MS cases, with some enrichment for subjects with primary progressive (PPMS) disease ( n  = 246) and 1210 healthy controls. Using logistic regression, we found that DRB1*1501 was strongly associated with risk ( P  = 7 × 10⁻⁴⁵), as expected, and after adjusting for DRB1*1501, a predisposing effect was also observed for DRB1*03 ( P  = 5 × 10⁻⁷). Individuals homozygous for either DRB1*15 or DRB1*03 were considerably more at risk of MS than heterozygotes and non-carriers. Both the DRB1*04 and the DRB1*01/DRB1*15 genotype combination, respectively, protected against PPMS in comparison to subjects with relapsing disease. Together, these data provide further evidence of heterogeneity at the DRB1 locus and confirm the importance of HLA variants in the phenotypic expression of MS.     

Dual luciferase labelling for non-invasive bioluminescence imaging of mesenchymal stromal cell chondrogenic differentiation in demineralized bone matrix scaffolds

Non-invasive bioluminescence imaging (BLI) to monitor changes in gene expression of cells implanted in live animals should facilitate the development of biomaterial scaffolds for tissue regeneration. We show that, in vitro, induction of chondrogenic differentiation in mouse bone marrow stromal cell line (CL1) and human adipose tissue derived mesenchymal stromal cells (hAMSCs), permanently transduced with a procollagen II (COL2A1) promoter driving a firefly luciferase gene reporter (PLuc) (COL2A1p·PLuc), induces PLuc expression in correlation with increases in COL2A1 and Sox9 mRNA expression and acquisition of chondrocytic phenotype. To be able to simultaneously monitor in vivo cell differentiation and proliferation, COL2A1p·PLuc labelled cells were also genetically labelled with a renilla luciferase (RLuc) gene driven by a constitutively active cytomegalovirus promoter, and then seeded in demineralized bone matrix (DBM) subcutaneously implanted in SCID mice. Non-invasive BLI monitoring of the implanted mice showed that the PLuc/RLuc ratio reports on gene expression changes indicative of cell differentiation. Large (CL1) and moderated (hAMSCs) changes in the PLuc/RLuc ratio over a 6 week period, revealed different patterns of in vivo chondrogenic differentiation for the CL1 cell line and primary MSCs, in agreement with in vitro published data and our results from histological analysis of DBM sections. This double bioluminescence labelling strategy together with BLI imaging to analyze behaviour of cells implanted in live animals should facilitate the development of progenitor cell/scaffold combinations for tissue repair.