Metagenomic deep sequencing of aqueous fluid detects intraocular lymphomas.

IntroductionCurrently, the detection of pathogens or mutations associated with intraocular lymphomas heavily relies on prespecified, directed PCRs. With metagenomic deep sequencing (MDS), an unbiased high-throughput sequencing approach, all pathogens as well as all mutations present in the host's genome can be detected in the same small amount of ocular fluid.MethodsIn this cross-sectional case series, aqueous fluid samples from two patients were submitted to MDS to identify pathogens as well as common and rare cancer mutations.ResultsMDS of aqueous fluid from the first patient with vitreal lymphoma revealed the presence of both Epstein-Barr virus (HHV-4/EBV) and human herpes virus 8 (HHV-8) RNA. Aqueous fluid from the second patient with intraocular B-cell lymphoma demonstrated a less common mutation in the MYD88 gene associated with B-cell lymphoma.ConclusionMDS detects pathogens that, in some instances, may drive the development of intraocular lymphomas. Moreover, MDS is able to identify both common and rare mutations associated with lymphomas

Timing of Sleep and Its Relationship with the Endogenous Melatonin Rhythm

While much research has investigated the effects of exogenous melatonin on sleep, less is known about the relationship between the timing of the endogenous melatonin rhythm and the sleep–wake cycle. Significant inter-individual variability in the phase relationship between sleep and melatonin rhythms has been reported although the extent to which the variability reflects intrinsic and/or environmental differences is unknown. We examined the effects of different sleeping schedules on the time of dim light melatonin onset (DLMO) in 28 young, healthy adults. Participants chose to maintain either an early (22:30–06:30 h) or a late (00:30–08:30 h) sleep schedule for at least 3 weeks prior to an overnight laboratory visit. Saliva samples were collected under dim light (<2 lux) and controlled posture conditions to determine salivary DLMO. The 2-h difference between groups in the enforced sleep–wake schedule was associated with a concomitant 1.75-h delay in DLMO. The mean phase relationship between sleep onset and DLMO remained constant (~2 h). The variance in DLMO time, however, was greater in the late group (range 4.5 h) compared to the early group (range 2.4 h) perhaps due to greater effect of environmental influences in delayed sleep types or greater intrinsic instability in their circadian system. The findings contribute to our understanding of individual differences in the human circadian clock and have important implications for the diagnosis and treatment of circadian rhythm sleep disorders, in particular if a greater normative range for phase angle of entrainment occurs in individuals with later sleep–wake schedules

ANNA: A UNIFIED FRAMEWORK FOR DATA SECURITY IN HEALTHCARE INTERNET OF THINGS

Today, the cyber-physical interactions are made possible with the development of internet and smart technologies. Smart objects such as sensors, RFID tags and actuators are the building blocks of such pervasive network, called as Internet of Things (IoT). IoT has a good number of real time applications in all domains including medical industry. It has great impact on modern healthcare with promising economic, technological and social benefits. Researchers across the globe already have started to explore a variety of technological solutions to enhance healthcare system by incorporating the potential of the IoT. This paper presents a unified framework ANNA, for integrating IoT in healthcare system. It presents three lightweight algorithms to enhance data security in the smart healthcare environment. In addition, this paper provides a detailed working scenario which would be helpful in implementing smart healthcare units in rural India. The working scenario of the ANNA smart healthcare system and the workflow of the framework are also elaborated in this paper. The proposed framework is proved against possible attacks in the healthcare environment. The salient features of the framework are outlined and this research creates a hope that the smart healthcare units based on ANNA framework will offer on time medical care to the needy and the downtrodden

The human ankyrin 1 promoter insulator sustains gene expression in a β-globin lentiviral vector in hematopoietic stem cells.

Lentiviral vectors designed for the treatment of the hemoglobinopathies require the inclusion of regulatory and strong enhancer elements to achieve sufficient expression of the β-globin transgene. Despite the inclusion of these elements, the efficacy of these vectors may be limited by transgene silencing due to the genomic environment surrounding the integration site. Barrier insulators can be used to give more consistent expression and resist silencing even with lower vector copies. Here, the barrier activity of an insulator element from the human ankyrin-1 gene was analyzed in a lentiviral vector carrying an antisickling human β-globin gene. Inclusion of a single copy of the Ankyrin insulator did not affect viral titer, and improved the consistency of expression from the vector in murine erythroleukemia cells. The presence of the Ankyrin insulator element did not change transgene expression in human hematopoietic cells in short-term erythroid culture or in vivo in primary murine transplants. However, analysis in secondary recipients showed that the lentiviral vector with the Ankyrin element preserved transgene expression, whereas expression from the vector lacking the Ankyrin insulator decreased in secondary recipients. These studies demonstrate that the Ankyrin insulator may improve long-term β-globin expression in hematopoietic stem cells for gene therapy of hemoglobinopathies

Creating New β-Globin-Expressing Lentiviral Vectors by High-Resolution Mapping of Locus Control Region Enhancer Sequences.

Hematopoietic stem cell gene therapy is a promising approach for treating disorders of the hematopoietic system. Identifying combinations of cis-regulatory elements that do not impede packaging or transduction efficiency when included in lentiviral vectors has proven challenging. In this study, we deploy LV-MPRA (lentiviral vector-based, massively parallel reporter assay), an approach that simultaneously analyzes thousands of synthetic DNA fragments in parallel to identify sequence-intrinsic and lineage-specific enhancer function at near-base-pair resolution. We demonstrate the power of LV-MPRA in elucidating the boundaries of previously unknown intrinsic enhancer sequences of the human β-globin locus control region. Our approach facilitated the rapid assembly of novel therapeutic βAS3-globin lentiviral vectors harboring strong lineage-specific recombinant control elements capable of correcting a mouse model of sickle cell disease. LV-MPRA can be used to map any genomic locus for enhancer activity and facilitates the rapid development of therapeutic vectors for treating disorders of the hematopoietic system or other specific tissues and cell types

Interakcije nekih plijesni i aflatoksinogenog soja Asspergillus flavus NRRL 3251

The objective of this study was to evaluate biotic interaction between some mould species and active producer of aflatoxin B1 Aspergillus flavus NRRL 3251, co-cultured in yeast-extract sucrose (YES) broth. Twenty-five mould strains of Alternaria spp., Cladosporium spp., Mucor spp., A. flavus and A. niger, used as biocompetitive agents, were isolated from outdoor and indoor airborne fungi, scrapings of mouldy household walls, and from stored and post-harvest maize. Aflatoxin B1 was extracted from mould biomasses with chloroform and detected using the multitoxin TLC method. The results confirm antagonistic interaction between all strains tested. With Alternaria spp. and Cladosporium spp., aflatoxin B1 production decreased 100 %, compared to detection in a single culture of A. flavus NRRL 3251 (Cmean=18.7 µg mL-1). In mixed cultures with Mucor spp., aflatoxin B1 levels dropped to (5.6-9.3) µg mL-1, and the inhibition was from 50 % to 70 %. Four of five aflatoxin non-producing strains of A. flavus interfered with aflatoxin production in mixed culture, and reduced AFB1 productivity by 100 %. One strain showed a lower efficacy in inhibiting AFB1 production (80 %) with a detectable amount of AFB1 3.7 µg mL-1 when compared to control. A decrease in toxin production was also observed in dual cultivation with A. niger strains. It resulted in 100 % reduction in three strains), 90 % reduction in one strain (Cmean=1.9 µg mL-1) and 80 % reduction in one strain (Cmean=3.7 µg mL-1) inhibition.Cilj rada bio je procijeniti biotske interakcije između sojeva različitih vrsta plijesni i kontrolnog soja Aspergillus flavus NRRL 3251, producenta aflatoksina B1 (AFB1). Inhibitorno djelovanje u miješanim kulturama na tvorbu AFB1 ispitano je na dvadeset pet sojeva Alternaria, Cladosporium, Mucor i Aspergillus vrsta izoliranih iz zraka, strugotina pljesnivih zidova te uskladištenog i prezimljenog kukuruza. Biosinteze su provedene u tekućoj hranjivoj podlozi s kvaščevim ekstraktom (YESbujon). Ekstrakcije AFB1 iz biomase izvršene su multitoksinskom metodom tankoslojne kromatografije. Rezultati biotskih interakcija pokazali su antagonistički odnos svih testiranih sojeva. Alternaria i Cladosporium vrste simultano inokulirane sporama A. flavus NRRL 3251 inhibirale su tvorbu AFB1 100 % u odnosu na dokazani toksin u kontrolnoj biosintezi (konc. 18,7 µg mL-1). U miješanim kulturama vrstama roda Mucor dokazane su padajuće koncentracije AFB1 (9,3 µg mL-1, 7,5 µg mL-1 i 5,6 µg mL-1), odnosno inhibicija tvorbe toksina 50 % do 70 %. Atoksinogeni sojevi A. flavus inhibirali su tvorbu AFB1 80 % (1 soj, konc. 3,7 µg mL-1) i 100 % (4 soja). Antagonističko djelovanje prema toksinogenom soju, smanjujući tvorbu AFB1 u rasponu 80 % do 100 % (konc. 1,9 µg mL-1 i 3,7 µg mL-1), dokazano je u uzgojnim biosintezama s A. niger

Publisher Correction: Structural Implications of Mutations Conferring Rifampin Resistance in Mycobacterium leprae.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has not been fixed in the paper

Correction to: Identification of novel Y chromosome encoded transcripts by testis transcriptome analysis of mice with deletions of the Y chromosome long arm.

Following publication of the original article [1], the following error was reported: The actin control panel in Fig. 3 of this paper is reproduced from Fig. 7 of Touré et al, 2004 [2] by kind permission of the Genetics Society of America. Touré et al, 2004 used Northern blotting to show that the Y-linked genes Ssty1 and Ssty2 have reduced expression in a range of mouse genotypes with deletions on the Y chromosome long arm. This paper shows that two novel genes, Sly and Asty are also present on mouse Yq and have reduced expression in these deleted genotypes. A further companion paper was published in Human Molecular Genetics (Ellis et al, 2005 [3]) showing that X-linked genes are upregulated in the various deleted genotypes. Since two of the genotypes concerned are sterile and very hard to generate, all the Northern blot experiments in these papers were performed on a single membrane that was stripped and re-probed with a range of different X- and Y-linked genes. The same beta-actin loading control image thus necessarily applies to all the data presented, and was shown in all three papers. We regret that this was not mentioned appropriately in the Methods and figure legends at the time of publication

A Nationally Representative Survey Assessing Restorative Sleep in US Adults

Restorative sleep is a commonly used term but a poorly defined construct. Few studies have assessed restorative sleep in nationally representative samples. We convened a panel of 7 expert physicians and researchers to evaluate and enhance available measures of restorative sleep. We then developed the revised Restorative Sleep Questionnaire (REST-Q), which comprises 9 items assessing feelings resulting from the prior sleep episode, each with 5-point Likert response scales. Finally, we assessed the prevalence of high, somewhat, and low REST-Q scores in a nationally representative sample of US adults (n= 1,055) and examined the relationship of REST-Q scores with other sleep and demographic characteristics. Pairwise correlations were performed between the REST-Q scores and other self-reported sleep measures. Weighted logistic regression analyses were conducted to compare scores on the REST-Q with demographic variables. The prevalence of higher REST-Q scores (4 or 5 on the Likert scale) was 28.1% in the nationally representative sample. REST-Q scores positively correlated with sleep quality (r=0.61) and sleep duration (r=0.32), and negatively correlated with both difficulty falling asleep (r=-0.40) and falling back asleep after waking (r=-0.41). Higher restorative sleep scores (indicating more feelings of restoration upon waking) were more common among those who were: ≥60 years of age (OR=4.20, 95%CI: 1.92-9.17); widowed (OR=2.35, 95%CI:1.01-5.42), and retired (OR=2.02, 95%CI:1.30-3.14). Higher restorative sleep scores were less frequent among those who were not working (OR=0.36, 95%CI: 0.10-1.00) and living in a household with two or more persons (OR=0.51,95%CI:0.29-0.87). Our findings suggest that the REST-Q may be useful for assessing restorative sleep