The case for early use of rapid whole-genome sequencing in management of critically ill infants: late diagnosis of Coffin-Siris syndrome in an infant with left congenital diaphragmatic hernia, congenital heart disease, and recurrent infections.

Congenital diaphragmatic hernia (CDH) results from incomplete formation of the diaphragm leading to herniation of abdominal organs into the thoracic cavity. CDH is associated with pulmonary hypoplasia, congenital heart disease, and pulmonary hypertension. Genetically, it is associated with aneuploidies, chromosomal copy-number variants, and single gene mutations. CDH is the most expensive noncardiac congenital defect. Management frequently requires implementation of extracorporeal membrane oxygenation (ECMO), which increases management expenditures 2.4-3.5-fold. The cost of management of CDH has been estimated to exceed $250 million per year. Despite in-hospital survival of 80%-90%, current management is imperfect, as a great proportion of surviving children have long-term functional deficits. We report the case of a premature infant prenatally diagnosed with CDH and congenital heart disease, who had a protracted and complicated course in the intensive care unit with multiple surgical interventions, including postcardiac surgery ECMO, gastrostomy tube placement with Nissen fundoplication, tracheostomy for respiratory failure, recurrent infections, and developmental delay. Rapid whole-genome sequencing (rWGS) identified a de novo, likely pathogenic, c.3096_ 3100delCAAAG (p.Lys1033Argfs*32) variant in ARID1B, providing a diagnosis of Coffin-Siris syndrome. Her parents elected palliative care and she died later that day

A Microfluidic Platform for Sequential Assembly and Separation of Synthetic Cell Models.

Funder: University of CambridgeFunder: Blavatnik Family FoundationCell-sized vesicles like giant unilamellar vesicles (GUVs) are established as a promising biomimetic model for studying cellular phenomena in isolation. However, the presence of residual components and byproducts, generated during vesicles preparation and manipulation, severely limits the utility of GUVs in applications like synthetic cells. Therefore, with the rapidly growing field of synthetic biology, there is an emergent demand for techniques that can continuously purify cell-like vesicles from diverse residues, while GUVs are being simultaneously synthesized and manipulated. We have developed a microfluidic platform capable of purifying GUVs through stream bifurcation, where a vesicles suspension is partitioned into three fractions: purified GUVs, residual components, and a washing solution. Using our purification approach, we show that giant vesicles can be separated from various residues─which range in size and chemical composition─with a very high efficiency (e = 0.99), based on size and deformability of the filtered objects. In addition, by incorporating the purification module with a microfluidic-based GUV-formation method, octanol-assisted liposome assembly (OLA), we established an integrated production-purification microfluidic unit that sequentially produces, manipulates, and purifies GUVs. We demonstrate the applicability of the integrated device to synthetic biology through sequentially fusing SUVs with freshly prepared GUVs and separating the fused GUVs from extraneous SUVs and oil droplets at the same time

Cow’s milk protein elimination in autistic children: language, cognitive and behavioral outcome

Background: Behavioral modification and structured education are necessary in autism but rather insufficient. Various dietary restrictions have been suggested as important prerequisites to benefit from other interventions in this disorder. Objective: This study was designed to highlight the degree of benefit in various aspects of development of autistic children upon elimination of cow's milk protein (CMP) from their diet and assess the level of specific IgE for CMP in their sera. Methods: The current study was conducted on 22 autistic children who were compared to 30 age and sex matched healthy children. Enrolled autistic children were randomly assigned to one of two groups. The parents of first group were instructed to eliminate cow milk (CM) from the diet of their children throughout the study period while patients of the second group were allowed to eat without restrictions. Each enrolled child was subjected to complete dietetic history taking, clinical examination and measurement of IgE for CM antigen in their sera by enzyme immunoassay. Autistic patients underwent a Childhood Autism Rating Scale (CARS) test. The patients were also subjected to language and intelligent quotient (IQ) testing, social and mental age assessment and child psychiatric evaluation. The autistic children received an interventional program for six months and were then re-evaluated using the previous clinical parameters. Results: The first group achieved significantly lower CARS test results (p < 0.01), significantly higher language age (p < 0.05) and significantly higher rate of change of CARS, language, social age, mental age and IQ (p < 0.001, <0.05, <0.05, <0.01 and <0.05 respectively) compared to the second group after 6 months of follow up. There was also a significantly higher mean specific IgE level to CMP in the autistic patients as compared to the controls. Additionally, 45.5% of patients who were on CM elimination diet went one CARS category down compared to only 36.4% of the second group. Conclusion: We report improvement in language, cognition and behavioral capabilities upon CM elimination in a group of autistic children. The higher CM specific IgE in these children may suggest that such adverse reaction to CM may have an allergic basis. Wider scale studies are needed to justify this adjuvant therapeutic option in autistic children hoping for better achievement from the current interventional programs. Keywords: Allergy – Autism – CARS – Cow milk – IgE – IQEgypt J Pediatr Allergy Immunol 2006; 4(1): 15-2

Direct Optofluidic Measurement of the Lipid Permeability of Fluoroquinolones.

Quantifying drug permeability across lipid membranes is crucial for drug development. In addition, reduced membrane permeability is a leading cause of antibiotic resistance in bacteria, and hence there is a need for new technologies that can quantify antibiotic transport across biological membranes. We recently developed an optofluidic assay that directly determines the permeability coefficient of autofluorescent drug molecules across lipid membranes. Using ultraviolet fluorescence microscopy, we directly track drug accumulation in giant lipid vesicles as they traverse a microfluidic device while exposed to the drug. Importantly, our measurement does not require the knowledge of the octanol partition coefficient of the drug - we directly determine the permeability coefficient for the specific drug-lipid system. In this work, we report measurements on a range of fluoroquinolone antibiotics and find that their pH dependent lipid permeability can span over two orders of magnitude. We describe various technical improvements for our assay, and provide a new graphical user interface for data analysis to make the technology easier to use for the wider community.The work was supported by an ERC Consolidator grant “DesignerPores” awarded to UFK. JC acknowledges support from the BBSRC. MS was supported by the Friedrich Naumann Foundation for Freedom and the Swiss- European Mobility Programme. KAN was supported by the Erasmus Plus student exchange programme. SHA is supported by a Herchel Smith Postdoctoral Fellowship. SP acknowledges support from the Leverhulme Trust through an Early Career Fellowship (ECF-2013-444).This is the final version of the article. It first appeared from Nature Publishing Group at http://dx.doi.org/10.1038/srep32824

Two Spectrophotometric Assays for Dopamine Derivatives in Pharmaceutical Products and in Biological Samples of Schizophrenic Patients Using Copper Tetramine Complex and Triiodide Reagent

Two simple, rapid, and sensitive spectrophotometric methods are proposed for the determination of levodopa (LD). The first method is based on coupling of 4-aminoantipyrine (4-AAP) with one of the dopamine derivatives (LD, CD) to give a new ligand that reacts with copper tetramine complex to give intensely colored chelates. The colored products are quantified spectrophotometrically at 525 and 520 nm for LD and CD, respectively. The optimization of the experimental conditions is described. The method has been used for the determination of 19.7–69.0 and 18.1–54.3 μg mL(−1) of LD and CD, respectively. The accuracy of the method is achieved by the values of recovery (100 ± 0.2%) and the precision is supported by the low standard deviation (SD = 0.17–0.59) and relative standard deviation (CV = 0.4%–1.54%) values. The second method is based on the formation of ion-pair iodinated inner sphere or outer sphere colored complexes between the LD and triiodide ions at pH 5 and room temperature (23 ± 3(°)C). This method has been used for the determination of LD within the concentration range 39.44–78.88 μg mL(−1) with SD = 0.22–0.24 and recovery percent = 100 ± 0.3%. The sensitivity of the two methods is indicated by Sandell's sensitivity of 0.014–0.019 g cm(−2). The results of the two methods are compared with those of the official method. The interference of common drug additives, degradation products, and excipients was also studied. The proposed methods were applied successfully to the determination of the LD-CD synthetic mixture and Levocare drug. The determination of LD in urine of some schizophrenic patients was applied with good precision and accuracy. The reliability of the methods was established by parallel determinations against the official British pharmacopoeia method

Uji Kualitas Briket Bioarang Berbahan Dasar Arang Kotoran Kambing, Arang Kotoran Sapi dan Arang Kotoran Ayam

The purpose of this study was to determine the quality of bioarang briquettes processed using goat manure charcoal, cow manure charcoal and chicken manure charcoal. This research will be carried out in December-February 2019, this research will be conducted in 2 locations, where in the first stage bioarang briquettes are made in the Laboratory of the Faculty of Agriculture Timor of University, while the second phase is tested on the quality of bioarang briquettes conducted at the Laboratory of the Faculty of Animal Husbandry, Nutrition and Science Feed Technology IPB. This study used a completely randomized design (CRD) consisting of three treatments: Goat manure briquette (GMB) : Goat droppings 90% + starch 10%; cow dung briquettes (CDB) : 90% Cow dung + 10% starch and ; chicken manure briquette (CMB): 90% chicken manure + 10% starch and repeated 10 times so that there are 30 units of experimental unit. Anova variance results showed that the parameters of ash content, combustion rate and heating value showed significant differences (P <0.05). It was concluded that the best quality briquettes are briquettes made from basic ingredients of goat manure which produce lower water content (36.12%), ash content (38.62%) and calorific value (3525 cal / g) while the best combustion rate is given by cow dung briquettes (0.29g / sec).Tujuan dari penelitian ini adalah untuk mengetahui kualitas briket bioarang yang diproses menggunakan arang kotoran kambing, arang kotoran sapi dan arang kotoran ayam. Penelitian ini dilaksanakan pada bulan Desember-Februari 2019, dan dilakukan di 2 lokasi, dimana pada tahap I dilakukan di Laboratorium Fakultas Pertanian Unimor untuk pembuatan briket bioarang, sedangkan pada tahap II dilakukan pengujian kualitas briket bioarang pada Laboratorium Fakultas Peternakan Departemen Ilmu Nutrisi dan Teknologi Pakan IPB. Penelitian ini menggunakan rancangan acak lengkap (RAL) yang terdiri dari tiga perlakuan : Briket kotoran kambing (BKK): kotoran Kambing 90 % + tepung kanji 10 %; briket kotoran sapi (BKS): kotoran Sapi 90 % + tepung kanji 10 % dan; briket kotoran ayam (BKA): kotoran Ayam 90 % + tepung kanji 10 % dan diulang sebanyak 10 kali sehingga terdapat 30 satuan unit percobaan. Hasil sidik ragam anova menunjukkan bahwa parameter kadar abu, laju pembakaran dan nilai kalor menunjukkan beda nyata (P<0,05). Disimpulkan bahwa kualitas briket terbaik adalah briket yang dibuat dari bahan dasar kotoran kambing yang menghasilkan kadar air lebih rendah (36,12 %), kadar abu (38,62 %) dan nilai kalor (3525 kal/g) sedangkan laju pembakaran terbaik diberikan oleh briket kotoran sapi (0,29 g/detik)

NASA Structural Analysis Report on the American Airlines Flight 587 Accident - Local Analysis of the Right Rear Lug

A detailed finite element analysis of the right rear lug of the American Airlines Flight 587 - Airbus A300-600R was performed as part of the National Transportation Safety Board s failure investigation of the accident that occurred on November 12, 2001. The loads experienced by the right rear lug are evaluated using global models of the vertical tail, local models near the right rear lug, and a global-local analysis procedure. The right rear lug was analyzed using two modeling approaches. In the first approach, solid-shell type modeling is used, and in the second approach, layered-shell type modeling is used. The solid-shell and the layered-shell modeling approaches were used in progressive failure analyses (PFA) to determine the load, mode, and location of failure in the right rear lug under loading representative of an Airbus certification test conducted in 1985 (the 1985-certification test). Both analyses were in excellent agreement with each other on the predicted failure loads, failure mode, and location of failure. The solid-shell type modeling was then used to analyze both a subcomponent test conducted by Airbus in 2003 (the 2003-subcomponent test) and the accident condition. Excellent agreement was observed between the analyses and the observed failures in both cases. From the analyses conducted and presented in this paper, the following conclusions were drawn. The moment, Mx (moment about the fuselage longitudinal axis), has significant effect on the failure load of the lugs. Higher absolute values of Mx give lower failure loads. The predicted load, mode, and location of the failure of the 1985-certification test, 2003-subcomponent test, and the accident condition are in very good agreement. This agreement suggests that the 1985-certification and 2003- subcomponent tests represent the accident condition accurately. The failure mode of the right rear lug for the 1985-certification test, 2003-subcomponent test, and the accident load case is identified as a cleavage-type failure. For the accident case, the predicted failure load for the right rear lug from the PFA is greater than 1.98 times the limit load of the lugs. I

Indole Pulse Signalling Regulates the Cytoplasmic pH of E. coli in a Memory-Like Manner.

Bacterial cells are critically dependent upon pH regulation. Here we demonstrate that indole plays a critical role in the regulation of the cytoplasmic pH of Escherichia coli. Indole is an aromatic molecule with diverse signalling roles. Two modes of indole signalling have been described: persistent and pulse signalling. The latter is illustrated by the brief but intense elevation of intracellular indole during stationary phase entry. We show that under conditions permitting indole production, cells maintain their cytoplasmic pH at 7.2. In contrast, under conditions where no indole is produced, the cytoplasmic pH is near 7.8. We demonstrate that pH regulation results from pulse, rather than persistent, indole signalling. Furthermore, we illustrate that the relevant property of indole in this context is its ability to conduct protons across the cytoplasmic membrane. Additionally, we show that the effect of the indole pulse that occurs normally during stationary phase entry in rich medium remains as a "memory" to maintain the cytoplasmic pH until entry into the next stationary phase. The indole-mediated reduction in cytoplasmic pH may explain why indole provides E. coli with a degree of protection against stresses, including some bactericidal antibiotics.The Leverhulme Trus

Assignment of PCR markers to river buffalo chromosomes

International audienc

Aerosol-jet printing facilitates the rapid prototyping of microfluidic devices with versatile geometries and precise channel functionalization.

Microfluidics has emerged as a powerful analytical tool for biology and biomedical research, with uses ranging from single-cell phenotyping to drug discovery and medical diagnostics, and only small sample volumes required for testing. The ability to rapidly prototype new designs is hugely beneficial in a research environment, but the high cost, slow turnaround, and wasteful nature of commonly used fabrication techniques, particularly for complex multi-layer geometries, severely impede the development process. In addition, microfluidic channels in most devices currently play a passive role and are typically used to direct flows. The ability to "functionalize" the channels with different materials in precise spatial locations would be a major advantage for a range of applications. This would involve incorporating functional materials directly within the channels that can partake in, guide or facilitate reactions in precisely controlled microenvironments. Here we demonstrate the use of Aerosol Jet Printing (AJP) to rapidly produce bespoke molds for microfluidic devices with a range of different geometries and precise "in-channel" functionalization. We show that such an advanced microscale additive manufacturing method can be used to rapidly design cost-efficient and customized microfluidic devices, with the ability to add functional coatings at specific locations within the microfluidic channels. We demonstrate the functionalization capabilities of our technique by specifically coating a section of a microfluidic channel with polyvinyl alcohol to render it hydrophilic. This versatile microfluidic device prototyping technique will be a powerful aid for biological and bio-medical research in both academic and industrial contexts