Novel LIPA mutations in Mexican siblings with lysosomal acid lipase deficiency

We would like to thank Radhika Tripuraneni, MD, MPH, for critical reading of the manuscript, Angelica TorizOrtiz, MD, for ultrasound imaging, and the medical staff of the Endoscopy and Pathology Department of CMN “20 de Noviembre”, along with all the personnel involved in the care of the patients. This work was presented in abstract form at 2013’s National Week of Gastroenterology (Semana Nacional de Gastroenterologia de la AMG) in Veracruz, Mexico and at Ⅸ Congress of SLEIMPN in Medellín, Colombia.Peer reviewedPublisher PD

Debt Counselling for Depression in Primary Care: an adaptive randomised controlled pilot trial (DeCoDer study)

Background: Depression and debt are common in the UK. The DeCoDer trial aimed to assess the clinical and cost effectiveness of the addition of Primary Care debt counselling advice service to usual care, for patients with depression and debt. However, the study was terminated early during the internal pilot trial phase because of recruitment delays. This report describes the rationale, methods and findings of the pilot study, and implications for future research. Objectives: The overarching aim of the internal pilot was to identify and resolve problems, thereby assessing the feasibility of the main trial. Specific objectives were to: confirm methods for practice recruitment, ability to recruit patients via the proposed approaches, determine acceptability of the study interventions and outcome measures; assess contamination, confirm the randomisation method for main trial, the level of participant attrition; and check robustness of data collection systems. Design: Adaptive parallel two group multi-centre randomised controlled pilot trial with nested mixed methods process and economic evaluation. Both individual and cluster (General Practice) level allocation were used in the pilot phase to assign participants to intervention or control groups. Setting: General practices in England and Wales.Participants: Individuals age ≥18 years, scoring ≥14 on the Beck Depression Inventory and self-identifying as having debt worries were included. Main exclusion criteria were: actively suicidal or psychotic and/or severely depressed and unresponsive to treatment, severe addiction to alcohol/illicit drugs, unable/unwilling to give written informed consent, currently participating in other research including follow-up phases, received Citizen’s Advice Bureau (CAB) debt advice in past year, and not wanting debt advice via GP practice. Interventions: Intervention: debt advice provided by CAB and shared biopsychosocial assessment in addition to treatment as usual (TAU) and two debt advice leaflets; Control: advice leaflets provided by GP and TAU only. Outcomes of pilot trial: Proportion of eligible patients who consented; number of participants recruited compared to target; assessment of contamination; assessment of patient satisfaction with intervention and outcome measures.Participant outcomes: Primary: Beck Depression Inventory II; Secondary: Psychological wellbeing, health and social care utilisation, service satisfaction, substance misuse, record of priority/non-priority debts, life events and difficulties and explanatory measures. Outcomes were assessed at baseline (pre-randomisation) and 4-months post randomisation. Other data sources: Qualitative interviews were conducted with participants, clinicians and CAB advisors.Results: Of the 238 expressions of interest screened, 61 participants (26%) were recruited and randomised (32 intervention and 29 control). All participants provided baseline outcomes and 52 provided primary outcome at four months follow up (14.7% drop out). 17 participants allocated to intervention saw CAB. Descriptive statistics are reported for participants with complete outcomes at baseline and 4-months’ follow up. Our qualitative findings suggest that the relationship between debt and depression is complex and the impact of each on the other is compounded by other psychological, social and contextual influences. Conclusions, Study Limitations and Future work: Due to low recruitment this trial was terminated at the internal pilot phase, and too small for inferential statistical analysis. We provide implications for conducting future research in this area

De novo DNA cytosine methyltransferase activities in mouse embryonic stem cells

It has been a controversial issue as to how many DNA cytosine methyltransferase mammalian cells have and whether de novo methylation and maintenance methylation activities are encoded by a single gene or two different genes. To address these questions, we have generated a null mutation of the only known mammalian DNA methyltransferase gene through homologous recombination in mouse embryonic stem cells and found that the development of the homozygous embryos is arrested prior to the 8-somite stage. Surprisingly, the null mutant embryonic stem cells are viable and contain low but stable levels of methyl cytosine and methyltransferase activity, suggesting the existence of a second DNA methyltransferase in mammalian cells. Further studies indicate that de novo methylation activity is not impaired by the mutation as integrated provirus DNA in MoMuLV-infected homozygous embryonic stem cells become methylated at a similar rate as in wild-type cells. Differentiation of mutant cells results in further reduction of methyl cytosine levels, consistent with the de novo methylation activity being down regulated in differentiated cells. These results provide the first evidence that an independently encoded DNA methyltransferase is present in mammalian cells which is capable of de novo methylating cellular and viral DNA in vivo

A Combined Modelling and Experimental Study of the Surface Energetics of a-Lactose Monohydrate

NoThe surface energy of a-lactose monohydrate measured by inverse gas chromatography (IGC) is reported along with a dynamic molecular modelling study of the interaction of the various molecular probes with different surfaces of a-lactose monohydrate. The IGC results show that a-lactose monohydrate is acidic in nature. Using quantitative calculations of the energy of adsorption, the acidic nature of the surface is confirmed and the calculated values agree closely with the experimentally measured values. Along with the acidic nature, dynamic molecular modelling also reveals that the presence of a channel and water molecules on a surface affects the surface energetics of that face. The presence of water on the surface can decrease or increase the surface energy by either blocking or attracting a probe molecule, respectively. This property of water depends on its position and association with other functional groups present on the surface. The effect of a channel or cavity on the surface energy is shown to depend on its size, which determines whether the functional groups in the channel are assessable by probe molecules or not. Overall molecular modelling explains, at the molecular level, the effect of different factors affecting the surface energy of individual faces of the crystal

Trp-dependent auxin biosynthesis in Arabidopsis: involvement of cytochrome P450s CYP79B2 and CYP79B3

The plant hormone auxin regulates many aspects of plant growth and development. Although several auxin biosynthetic pathways have been proposed, none of these pathways has been precisely defined at the molecular level. Here we provide in planta evidence that the two Arabidopsis cytochrome P450s, CYP79B2 and CYP79B3, which convert tryptophan (Trp) to indole-3-acetaldoxime (IAOx) in vitro, are critical enzymes in auxin biosynthesis in vivo. IAOx is thus implicated as an important intermediate in auxin biosynthesis. Plants overexpressing CYP79B2 contain elevated levels of free auxin and display auxin overproduction phenotypes. Conversely, cyp79B2 cyp79B3 double mutants have reduced levels of IAA and show growth defects consistent with partial auxin deficiency. Together with previous work on YUCCA, a flavin monooxygenase also implicated in IAOx production, and nitrilases that convert indole-3-acetonitrile to auxin, this work provides a framework for further dissecting auxin biosynthetic pathways and their regulation

Activin receptor-like kinase 1 modulates transforming growth factor-β1 signaling in the regulation of angiogenesis

The activin receptor-like kinase 1 (ALK1) is a type I receptor for transforming growth factor-β (TGF-β) family proteins. Expression of ALK1 in blood vessels and mutations of the ALK1 gene in human type II hereditary hemorrhagic telangiectasia patients suggest that ALK1 may have an important role during vascular development. To define the function of ALK1 during development, we inactivated the ALK1 gene in mice by gene targeting. The ALK1 homozygous embryos die at midgestation, exhibiting severe vascular abnormalities characterized by excessive fusion of capillary plexes into cavernous vessels and hyperdilation of large vessels. These vascular defects are associated with enhanced expression of angiogenic factors and proteases and are characterized by deficient differentiation and recruitment of vascular smooth muscle cells. The blood vessel defects in ALK1-deficient mice are reminiscent of mice lacking TGF-β1, TGF-β type II receptor (TβR-II), or endoglin, suggesting that ALK1 may mediate TGF-β1 signal in endothelial cells. Consistent with this hypothesis, we demonstrate that ALK1 in endothelial cells binds to TGF-β1 and TβR-II. Furthermore, the ALK1 signaling pathway can inhibit TGF-β1-dependent transcriptional activation mediated by the known TGF-β1 type I receptor, ALK5. Taken together, our results suggest that the balance between the ALK1 and ALK5 signaling pathways in endothelial cells plays a crucial role in determining vascular endothelial properties during angiogenesis