Pre-incubation of cell-free HIV-1 group M isolates with non-nucleoside reverse transcriptase inhibitors blocks subsequent viral replication in co-cultures of dendritic cells and T cells.

In order to study the inhibitory effect of various reverse transcriptase inhibitors (RTIs) on cell-free HIV, we adapted a recently described in vitro system, based on co-cultures of dendritic cells and resting CD4 T cells, modelling early target cells during sexual transmission. The compounds tested included the second-generation non-nucleoside RTI (NNRTI) TMC-120 (R147681, dapivirine) and TMC-125 (R165335, travertine), as well as the reference nucleoside RTI AZT (zidovudine), the nucleotide RTI PMPA (tenofovir) and the NNRTI UC-781. The virus strains included the reference strain HIV-1Ba-L and six primary isolates, representative of the HIV-1 group M pandemic. They all display the non-syncytium-inducing and CCR5 receptor-using (NSI/R5) phenotype, important in transmission. Cell-free virus was immobilized on a poly-L-lysine (PLL)-treated microwell plate and incubated with compound for 1 h. Afterwards, the compound was thoroughly washed away; target cells were added and cultured for 2 weeks, followed by an extended culture with highly susceptible mitogen-activated T cells. Viral production in the cultures was measured on supernatant with HIV antigen ELISA. Negative results were confirmed by showing absence of proviral DNA in the cells. TMC-120 and TMC-125 inhibited replication of HIV-1Ba-L with average EC50 values of 38 nM and 117 nM, respectively, whereas the EC50 of UC-781 was 517 nM. Complete suppression of virus and provirus was observed at compound concentrations of 100, 300 and 1000 nM, respectively. Inhibition of all primary isolates followed the same pattern as HIV-1Ba-L. In contrast, pre-treating the virus with the nucleotide RTI PMPA and AZT failed to inhibit infection even at a concentration of 100000 nM. These data clearly suggest that NNRTIs inactivate RT enzymatic activity of different viral clades (predominant in the epidemic) and might be proposed for further testing as a sterilizing microbicide worldwide

Normal mucus formation requires cAMP-dependent HCO3- secretion and Ca2+-mediated mucin exocytosis.

  Evidence from the pathology in cystic fibrosis (CF) and recent results in vitro indicate that HCO3- is required for gel-forming mucins to form the mucus that protects epithelial surfaces. Mucus formation and release is a complex process that begins with an initial intracellular phase of synthesis, packaging and apical granule exocytosis that is followed by an extracellular phase of mucin swelling, transport and discharge into a lumen. Exactly where HCO3- becomes crucial in these processes is unknown, but we observed that in the presence of HCO3-, stimulating dissected segments of native mouse intestine with 5-hydroxytryptamine (5-HT) and prostaglandin E2 (PGE2) induced goblet cell exocytosis followed by normal mucin discharge in wild-type (WT) intestines. CF intestines that inherently lack cystic fibrosis transmembrane conductance regulator (CFTR)-dependent HCO3- secretion also demonstrated apparently normal goblet cell exocytosis, but in contrast, this was not followed by similar mucin discharge. Moreover, we found that even in the presence of HCO3-, when WT intestines were stimulated only with a Ca2+-mediated agonist (carbachol), exocytosis was followed by poor discharge as with CF intestines. However, when the Ca2+-mediated agonist was combined with a cAMP-mediated agonist (isoproterenol (isoprenaline) or vasoactive intestinal peptide) in the presence of HCO3- both normal exocytosis and normal discharge was observed. These results indicate that normal mucus formation requires concurrent activation of a Ca2+-mediated exocytosis of mucin granules and an independent cAMP-mediated, CFTR-dependent, HCO3- secretion that appears to mainly enhance the extracellular phases of mucus excretion

Dynamical Evolution of Globular Cluster Systems formed in Galaxy Mergers: Deep HST/ACS Imaging of Old and Intermediate-Age Globular Clusters in NGC 3610

(ABRIDGED) The ACS camera on board the Hubble Space Telescope has been used to obtain deep images of the giant elliptical galaxy NGC 3610, a well-established dissipative galaxy merger remnant. These observations supersede previous WFPC2 images which revealed the presence of a population of metal-rich globular clusters (GCs) of intermediate age (~1.5-4 Gyr). We detect a total of 580 GC candidates, 46% more than from the previous WFPC2 images. The new photometry strengthens the significance of the previously found bimodality of the color distribution of GCs. Peak colors in V-I are 0.93 +/-0.01 and 1.09 +/- 0.01 for the blue and red subpopulations, respectively. The luminosity function (LF) of the inner 50% of the metal-rich (`red') population of GCs differs markedly from that of the outer 50%. In particular, the LF of the inner 50% of the red GCs shows a flattening consistent with a turnover that is about 1.0 mag fainter than the turnover of the blue GC LF. This is consistent with predictions of recent models of GC disruption for the age range mentioned above and for metallicities that are consistent with the peak color of the red GCs as predicted by population synthesis models. We determine the specific frequency of GCs in NGC 3610 and find a present-day value of S_N = 1.4 +/- 0.6. We estimate that this value will increase to S_N = 3.8 +/- 1.7 at an age of 10 Gyr, which is consistent with typical S_N values for `normal' ellipticals. Our findings constitute further evidence in support of the notion that metal-rich GC populations formed during major mergers involving gas-rich galaxies can evolve dynamically (through disruption processes) into the red, metal-rich GC populations that are ubiquitous in `normal' giant ellipticals.Comment: 15 pages, 14 figures, 4 tables. Accepted for publication in The Astronomical Journal. Figure 6 somewhat degraded to adhere to astro-ph rule

A Catalog of Transient X-ray Sources in M31

From October 1999 to August 2002, 45 transient X-ray sources were detected in M31 by Chandra and XMM-Newton. We have performed spectral analysis of all XMM-Newton and Chandra ACIS detections of these sources, as well as flux measurements of Chandra HRC detections. The result is absorption-corrected X-ray lightcurves for these sources covering this 2.8 year period, along with spectral parameters for several epochs of the outbursts of most of the transient sources. We supply a catalog of the locations, outburst dates, peak observed luminosities, decay time estimates, and spectral properties of the transient sources, and we discuss similarities with Galactic X-ray novae. Duty cycle estimates are possible for 8 of the transients and range from 40% to 2%; upper limits to the duty cycles are estimated for an additional 15 transients and cover a similar range. We find 5 transients which have rapid decay times and may be ultra-compact X-ray binaries. Spectra of three of the transients suggest they may be faint Galactic foreground sources. If even one is a foreground source, this suggests a surface density of faint transient X-ray sources of >~1 deg$^{-2}$.Comment: 63 pages, 22 figures, 3 tables, accepted for publication in Ap

Cyclosporine a Alters Expression of Renal Micrornas: New Insights into Calcineurin Inhibitor Nephrotoxicity

Calcineurin inhibitors are powerful immunosuppressants that revolutionized organ transplantation. However, non-immune effects of the calcineurin inhibitor, such as cyclosporine A (CsA), have significantly hindered their use. Specifically, nephrotoxicity, which is associated with tubulointerstitial fibrosis, inflammation, and podocyte damage, affects up to half of all transplant patients. Calcineurin is involved in many aspects of kidney development and function; therefore, mechanisms of CsA-induced nephrotoxicity are complex and not yet fully understood. MicroRNAs are short non-coding RNAs that regulate protein-coding RNA expression through post-translational repression of target messenger RNAs. MicroRNA dysregulation is known to be involved in kidney diseases including fibrosis. In this study, we compared the renal microRNA expression profiles between mice that received CsA (20 mg/kg) or vehicle daily for six weeks. The results demonstrate that CsA induces significant changes in renal microRNA expression profile. We used combined criteria of False Discovery Rate (≤0.1), fold change (≥2) and median signal strength (≥50) and identified 76 differencially expressed microRNAs. This approach identified microRNAs previously linked to renal fibrosis that includes let-7d, miR-21, miR-29, miR-30, miR-130, miR-192, and miR-200 as well as microRNAs that have not been reported to be related to nephrotoxicity or immunosuppression. Pathway analysis of microRNA/mRNA changes highlights the Wnt, TGF-β, mTOR, and VEGF pathways. The mRNA expression profiles were compared in the same samples. The change of mRNA and microRNA profiles showed close correlations. To validate that the observed microRNA and mRNA expression level changes in mice kidney tissue were directly related to CsA treatment, the expression change induced by CsA treatment of three microRNAs (miR-21, miR-186, and miR-709) and three mRNAs (BMPR1a, SMURF1 and SMAD7) were compared in HEK293 cell line. A similar trend of expression level change was induced by CsA treatment in all selected microRNAs and mRNAs in the in vitro cell model. These data provide a roadmap for future work to study the role of the known and novel candidate microRNAs in the mechanism of nephrotoxicity and their further therapeutic potential

Sacral agenesis: a pilot whole exome sequencing and copy number study

Background: Caudal regression syndrome (CRS) or sacral agenesis is a rare congenital disorder characterized by a constellation of congenital caudal anomalies affecting the caudal spine and spinal cord, the hindgut, the urogenital system, and the lower limbs. CRS is a complex condition, attributed to an abnormal development of the caudal mesoderm, likely caused by the effect of interacting genetic and environmental factors. A well-known risk factor is maternal type 1 diabetes. Method: Whole exome sequencing and copy number variation (CNV) analyses were conducted on 4 Caucasian trios to identify de novo and inherited rare mutations. Results: In this pilot study, exome sequencing and copy number variation (CNV) analyses implicate a number of candidate genes, including SPTBN5, MORN1, ZNF330, CLTCL1 and PDZD2. De novo mutations were found in SPTBN5, MORN1 and ZNF330 and inherited predicted damaging mutations in PDZD2 (homozygous) and CLTCL1 (compound heterozygous). Importantly, predicted damaging mutations in PTEN (heterozygous), in its direct regulator GLTSCR2 (compound heterozygous) and in VANGL1 (heterozygous) were identified. These genes had previously been linked with the CRS phenotype. Two CNV deletions, one de novo (chr3q13.13) and one homozygous (chr8p23.2), were detected in one of our CRS patients. These deletions overlapped with CNVs previously reported in patients with similar phenotype. Conclusion: Despite the genetic diversity and the complexity of the phenotype, this pilot study identified genetic features common across CRS patients

Genetic study of congenital bile-duct dilatation identifies de novo and inherited variants in functionally related genes

Background: Congenital dilatation of the bile-duct (CDD) is a rare, mostly sporadic, disorder that results in bile retention with severe associated complications. CDD affects mainly Asians. To our knowledge, no genetic study has ever been conducted. Methods: We aim to identify genetic risk factors by a “trio-based” exome-sequencing approach, whereby 31 CDD probands and their unaffected parents were exome-sequenced. Seven-hundred controls from the local population were used to detect gene-sets significantly enriched with rare variants in CDD patients. Results: Twenty-one predicted damaging de novo variants (DNVs; 4 protein truncating and 17 missense) were identified in several evolutionarily constrained genes (p < 0.01). Six genes carrying DNVs were associated with human developmental disorders involving epithelial, connective or bone morphologies (PXDN, RTEL1, ANKRD11, MAP2K1, CYLD, ACAN) and four linked with cholangio- and hepatocellular carcinomas (PIK3CA, TLN1 CYLD, MAP2K1). Importantly, CDD patients have an excess of DNVs in cancer-related genes (p < 0.025). Thirteen genes were recurrently mutated at different sites, forming compound heterozygotes or functionally related complexes within patients. Conclusions: Our data supports a strong genetic basis for CDD and show that CDD is not only genetically heterogeneous but also non-monogenic, requiring mutations in more than one genes for the disease to develop. The data is consistent with the rarity and sporadic presentation of CDD

ONOMATOPOEIA: LIFE EXPERIENCES OF POETS ON EXPRESSING AGGRESSION

People engaged in writing and performing poetry as a way of expressing their emotions is a rising phenomenon. The study aimed to know the life experiences of poets in coping with aggression through poetry writing. As well as to know if there is a significant data that will prove the existence of the aggression that motivates poets to write. The study utilized the qualitative design it’s an inquiry process of understanding based on particular distinct methodological traditions on request that investigate a social or human problem, under the Qualitative design the researchers used the Case Study method The respondents of the study were two female and two male poetry writers, 18 years old and above and who writes poetry for at least 3 years, and has an average deviation / average score from the Buss-Perry Aggression test. The researchers conducted the study on different cities in the National Capital Region. The researchers were able to come up with the following themes; (1) Personal experiences (2) Social experiences (3) Coping mechanisms. These themes shows the lived experiences of the poets in dealing with aggression by using poetry as a mechanism in coping up with the different aspects of life that significantly contributes to their aggression. We recommend for future researchers to utilize different approaches and expand the scope of this research.&nbsp