SPEN haploinsufficiency causes a neurodevelopmental disorder overlapping proximal 1p36 deletion syndrome with an episignature of X chromosomes in females.

Deletion 1p36 (del1p36) syndrome is the most common human disorder resulting from a terminal autosomal deletion. This condition is molecularly and clinically heterogeneous. Deletions involving two non-overlapping regions, known as the distal (telomeric) and proximal (centromeric) critical regions, are sufficient to cause the majority of the recurrent clinical features, although with different facial features and dysmorphisms. SPEN encodes a transcriptional repressor commonly deleted in proximal del1p36 syndrome and is located centromeric to the proximal 1p36 critical region. Here, we used clinical data from 34 individuals with truncating variants in SPEN to define a neurodevelopmental disorder presenting with features that overlap considerably with those of proximal del1p36 syndrome. The clinical profile of this disease includes developmental delay/intellectual disability, autism spectrum disorder, anxiety, aggressive behavior, attention deficit disorder, hypotonia, brain and spine anomalies, congenital heart defects, high/narrow palate, facial dysmorphisms, and obesity/increased BMI, especially in females. SPEN also emerges as a relevant gene for del1p36 syndrome by co-expression analyses. Finally, we show that haploinsufficiency of SPEN is associated with a distinctive DNA methylation episignature of the X chromosome in affected females, providing further evidence of a specific contribution of the protein to the epigenetic control of this chromosome, and a paradigm of an X chromosome-specific episignature that classifies syndromic traits. We conclude that SPEN is required for multiple developmental processes and SPEN haploinsufficiency is a major contributor to a disorder associated with deletions centromeric to the previously established 1p36 critical regions.

Duloxetine as an Analgesic in Patients Who Do Not Have Central Sensitivity Undergoing Single-Setting, Bilateral Total Knee Arthroplasty: A Prospective, Double-Blinded, Randomized, Placebo-Controlled Trial.

BACKGROUND: Pain control and patient satisfaction after total knee arthroplasty (TKA) have room for improvement. While studies have reported better analgesic outcomes with antidepressants like duloxetine in patients who do not have central sensitivity (CS), we undertook this trial to determine the short and midterm analgesic role of low-dose duloxetine in patients who do not have CS. METHODS: This prospective, double-blinded, randomized, placebo-controlled trial was conducted in 106 patients undergoing single-setting, bilateral TKA under spinal anesthesia. There were 2 matched groups, with one given 20 mg of duloxetine and the other given a placebo (similar in appearance and weight) from preoperative day 2 to postoperative day 28. Follow-ups were scheduled at 48-hours, 1-week, 2-weeks, 4-weeks, and 3-months. Pain was measured using a visual analogue scale at rest and visual analogue scale at mobilization (mVAS). Secondary measures included additional non-steroidal anti-inflammatory drug consumption, patient satisfaction, and safety profile. RESULTS: The visual analogue scale at rest in the duloxetine group was better in the first 48 hours (6.38 ± 1.32 versus 7.02 ± 0.99; P = .017), 1-week (4.76 ± 1.24 versus 5.89 ± 1.06; P < .001), and 2-weeks (3.34 ± 1.19 versus 4.26 ± 1.02; P < .001) follow-up. The mVAS remained significantly higher in the duloxetine group in the first 48 hours (7.23 ± 1.12 versus 8.21 ± 0.69; P < .001), 1-week (5.83 ± 1.11 versus 6.82 ± 0.92; P < .001), and 2 weeks (3.70 ± 0.89 versus 4.60 ± 1.03; P < .001) follow-up. Both outcomes became comparable from 4-week follow-up onward. Patient satisfaction (8.44 ± 1.68 versus 7.17 ± 1.04; P < .001) and additional non-steroidal anti-inflammatory drug consumption (2,770 ± 533.05 versus 3,566.04 ± 464.54; P < .001) were better in the duloxetine group, with a comparable safety profile. CONCLUSIONS: In patients who did not have CS, persistent pain after bilateral TKA can be managed safely and successfully by a daily dose of 20 mg Duloxetine, improving patient satisfaction and analgesic consumption in the acute postoperative phase.

An intrinsically disordered region of methyl-CpG binding domain protein 2 (MBD2) recruits the histone deacetylase core of the NuRD complex.

The MBD2-NuRD (Nucleosome Remodeling and Deacetylase) complex is an epigenetic reader of DNA methylation that regulates genes involved in normal development and neoplastic diseases. To delineate the architecture and functional interactions of the MBD2-NuRD complex, we previously solved the structures of MBD2 bound to methylated DNA and a coiled-coil interaction between MBD2 and p66α that recruits the CHD4 nucleosome remodeling protein to the complex. The work presented here identifies novel structural and functional features of a previously uncharacterized domain of MBD2 (MBD2IDR). Biophysical analyses show that the MBD2IDR is an intrinsically disordered region (IDR). However, despite this inherent disorder, MBD2IDR increases the overall binding affinity of MBD2 for methylated DNA. MBD2IDR also recruits the histone deacetylase core components (RbAp48, HDAC2 and MTA2) of NuRD through a critical contact region requiring two contiguous amino acid residues, Arg(286) and Leu(287). Mutating these residues abrogates interaction of MBD2 with the histone deacetylase core and impairs the ability of MBD2 to repress the methylated tumor suppressor gene PRSS8 in MDA-MB-435 breast cancer cells. These findings expand our knowledge of the multi-dimensional interactions of the MBD2-NuRD complex that govern its function.

Mi2ß-mediated silencing of the fetal γ-globin gene in adult erythroid cells.

An understanding of the human fetal to adult hemoglobin switch offers the potential to ameliorate ß-type globin gene disorders such as sickle cell anemia and ß-thalassemia through activation of the fetal γ-globin gene. Chromatin modifying complexes, including MBD2-NuRD and GATA-1/FOG-1/NuRD, play a role in γ-globin gene silencing, and Mi2ß (CHD4) is a critical component of NuRD complexes. We observed that knockdown of Mi2ß relieves γ-globin gene silencing in ß-YAC transgenic murine chemical inducer of dimerization hematopoietic cells and in CD34(+) progenitor-derived human primary adult erythroid cells. We show that independent of MBD2-NuRD and GATA-1/FOG-1/NuRD, Mi2ß binds directly to and positively regulates both the KLF1 and BCL11A genes, which encode transcription factors critical for γ-globin gene silencing during ß-type globin gene switching. Remarkably, <50% knockdown of Mi2ß is sufficient to significantly induce γ-globin gene expression without disrupting erythroid differentiation of primary human CD34(+) progenitors. These results indicate that Mi2ß is a potential target for therapeutic induction of fetal hemoglobin.

p66Alpha-MBD2 coiled-coil interaction and recruitment of Mi-2 are critical for globin gene silencing by the MBD2-NuRD complex.

Nucleosome remodeling complexes comprise several large families of chromatin modifiers that integrate multiple epigenetic control signals to play key roles in cell type-specific transcription regulation. We previously isolated a methyl-binding domain protein 2 (MBD2)-containing nucleosome remodeling and deacetylation (NuRD) complex from primary erythroid cells and showed that MBD2 contributes to DNA methylation-dependent embryonic and fetal ß-type globin gene silencing during development in vivo. Here we present structural and biophysical details of the coiled-coil interaction between MBD2 and p66α, a critical component of the MBD2-NuRD complex. We show that enforced expression of the isolated p66α coiled-coil domain relieves MBD2-mediated globin gene silencing and that the expressed peptide interacts only with a subset of components of the MBD2-NuRD complex that does not include native p66α or Mi-2. These results demonstrate the central importance of the coiled-coil interaction and suggest that MBD2-dependent DNA methylation-driven gene silencing can be disrupted by selectively targeting this coiled-coil complex.

Immobilization of cellulase on iron tolerant Pseudomonas stutzeri biosynthesized photocatalytically active magnetic nanoparticles for increased thermal stability.

Bacteria mediated synthesis of magnetic nanoparticles (MNPs) for biotechnological applications is an important area of nanotechnology. This study demonstrates the use of iron tolerant bacterium for synthesis of MNPs for cellulase immobilization and photocatalytic activity. The enrichment, isolation, screening and molecular identification led to the selection of Pseudomonas stutzeri KDP_M2 with high degree of iron tolerance. The synthesis parameters such as 1 mM ferric quinate, pH 9 and 96 h static incubation were found optimum for maximum yield of 210 mg/L. The characterization using various techniques indicated that MNPs were Hematite (Fe2O3) with particle size between 10 and 20 nm. Further, vibrating sample magnetometer and thermogravimetric analyses demonstrated the superparamagnetic nature with high thermal stability. The MNPs were found an excellent support for immobilization of industrially important cellulase with 96.5% binding efficiency. The immobilization which was confirmed by Fourier transform infrared spectroscopy indicated that immobilization did not reduce the cellulase activity, rather enhanced the thermal stability and operational temperature range of cellulase. The immobilized cellulase showed maximum cellulolytic activity at pH 4.6 and retained 80% activity upto 3rd cycle of reuse, therefore, can be utilized repeatedly at acidic conditions.The monitoring the photocatalytic activity showed rapid degradation of methyl violet and methylene blue within initial 10 min. of reaction.

Selective and sensitive colorimetric detection of platinum using Pseudomonas stutzeri mediated optimally synthesized antibacterial silver nanoparticles.

In this work, Pseudomonas stutzeri was used for the optimum biogenic synthesis of antibacterial silver nanoparticles (AgNPs) which were applied for colorimetric detection of platinum ions (Pt+2). The optimum synthesis conditions were 2 mM AgNO3, pH 9 and incubation at 60 °C for 24 h. The FTIR spectra indicated that biomolecules such as amino acids, proteins or enzymes from P. stutzeri were involved in the synthesis of AgNPs in the size range of 10-50 nm. Among the various metal ions tested and screened initially, the colloidal AgNPs probe-based colorimetric assay selectively detected Pt+2 with 50 ppm as the limit of detection (LOD). The assay demonstrated in the present study quantitatively recovered Pt+2 in the range of 70-150 % with good accuracy and precision. Further, the test of antibacterial activity of AgNPs alone, and in combination with ampicillin showed excellent activity against four of the six tested bacteria.

Selective interaction between phytomediated anionic silver nanoparticles and mercury leading to amalgam formation enables highly sensitive, colorimetric and memristor-based detection of mercury.

Presently, nanotechnology is being foreseen to play an important role in developing analytical assays for the detection of pollutants like mercury (Hg2+). In this study, Kokum fruit mediated silver nanoparticles (AgNPs) were differentially centrifuged to prepare anionic, monodispersed AgNPs to develop a highly sensitive, colorimetric and memristor-based assay for detection of Hg2+ in water samples. The investigation of the highly selective reaction between AgNPs and Hg2+ using HAADF-STEM images and EDS spectrum indicated the amalgam formation through etching and under potential deposition which resulted in a visible color change from brown to colorless, change in SPR intensity and also change in memristive switching like property of AgNPs. The developed colorimetric assay detected Hg2+ with a limit of detection (LOD) of 6.2 ppb and limit of quantification (LOQ) of 18.9 ppb and, quantitatively recovered Hg2+ with good accuracy and precision (RSD < 2%). Further, the test of memristive switching like property of AgNPs demonstrated frequency-dependent shrinkage of I-V hysteresis loop indicating memristive switching like property. The test of the sensitivity of Hg2+ detection was estimated to be 8.7 ppb as the LOD and 26.4 ppb as LOQ. Like the colorimetric assay, the memristor-based assay also recovered Hg2+ with good accuracy and precision.

Managing highly insulin-resistant diabetes mellitus: weight loss approaches and medical management.

The prevalence of obesity and diabetes is epidemic. Severe insulin resistance (defined as the need for > or = 200 units of insulin per day to achieve glycemic control) is commonly seen with obesity and can complicate diabetes management. The management of patients with diabetes who have severe insulin resistance is difficult, and at times frustrating, and requires a multifaceted approach. Weight loss is the best treatment option, which can be a challenging task for patients to achieve and maintain. Medications that decrease insulin needs like metformin, thiazolidinediones, or pramlintide may help, but some patients also need high doses of insulin. This article reviews these different treatment options and provides practical advice on weight loss, use of insulin sensitizers, and use of U-500 insulin.