A G protein γ subunit-like domain shared between RGS11 and other RGS proteins specifies binding to Gβ5 subunits

Regulators of G protein signaling (RGS) proteins act as GTPase-activating proteins (GAPs) toward the α subunits of heterotrimeric, signal-transducing G proteins. RGS11 contains a G protein γ subunit-like (GGL) domain between its Dishevelled/Egl-10/Pleckstrin and RGS domains. GGL domains are also found in RGS6, RGS7, RGS9, and the Caenorhabditis elegans protein EGL-10. Coexpression of RGS11 with different Gβ subunits reveals specific interaction between RGS11 and Gβ5. The expression of mRNA for RGS11 and Gβ5 in human tissues overlaps. The Gβ5/RGS11 heterodimer acts as a GAP on Gαo, apparently selectively. RGS proteins that contain GGL domains appear to act as GAPs for Gα proteins and form complexes with specific Gβ subunits, adding to the combinatorial complexity of G protein-mediated signaling pathways.

The GTPase activating factor for transducin in rod photoreceptors is the complex between RGS9 and type 5 G protein β subunit

Proteins of the regulators of G protein signaling (RGS) family modulate the duration of intracellular signaling by stimulating the GTPase activity of G protein α subunits. It has been established that the ninth member of the RGS family (RGS9) participates in accelerating the GTPase activity of the photoreceptor-specific G protein, transducin. This process is essential for timely inactivation of the phototransduction cascade during the recovery from a photoresponse. Here we report that functionally active RGS9 from vertebrate photoreceptors exists as a tight complex with the long splice variant of the G protein β subunit (Gβ5L). RGS9 and Gβ5L also form a complex when coexpressed in cell culture. Our data are consistent with the recent observation that several RGS proteins, including RGS9, contain G protein γ-subunit like domain that can mediate their association with Gβ5 (Snow, B. E., Krumins, A. M., Brothers, G. M., Lee, S. F., Wall, M. A., Chung, S., Mangion, J., Arya, S., Gilman, A. G. & Siderovski, D. P. (1998) Proc. Natl. Acad. Sci. USA 95, 13307–13312). We report an example of such a complex whose cellular localization and function are clearly defined.

Mutations in the chloride-bicarbonate exchanger gene AE1 cause autosomal dominant but not autosomal recessive distal renal tubular acidosis

Primary distal renal tubular acidosis (dRTA) is characterized by reduced ability to acidify urine, variable hyperchloremic hypokalemic metabolic acidosis, nephrocalcinosis, and nephrolithiasis. Kindreds showing either autosomal dominant or recessive transmission are described. Mutations in the chloride-bicarbonate exchanger AE1 have recently been reported in four autosomal dominant dRTA kindreds, three of these altering codon Arg589. We have screened 26 kindreds with primary dRTA for mutations in AE1. Inheritance was autosomal recessive in seventeen kindreds, autosomal dominant in one, and uncertain due to unknown parental phenotype or sporadic disease in eight kindreds. No mutations in AE1 were detected in any of the autosomal recessive kindreds, and analysis of linkage showed no evidence of linkage of recessive dRTA to AE1. In contrast, heterozygous mutations in AE1 were identified in the one known dominant dRTA kindred, in one sporadic case, and one kindred with two affected brothers. In the dominant kindred, the mutation Arg-589/Ser cosegregated with dRTA in the extended pedigree. An Arg-589/His mutation in the sporadic case proved to be a de novo mutation. In the third kindred, affected brothers both have an intragenic 13-bp duplication resulting in deletion of the last 11 amino acids of AE1. These mutations were not detected in 80 alleles from unrelated normal individuals. These findings underscore the key role of Arg-589 and the C terminus in normal AE1 function, and indicate that while mutations in AE1 cause autosomal dominant dRTA, defects in this gene are not responsible for recessive disease.

Structure-assisted design of mechanism-based irreversible inhibitors of human rhinovirus 3C protease with potent antiviral activity against multiple rhinovirus serotypes

Human rhinoviruses, the most important etiologic agents of the common cold, are messenger-active single-stranded monocistronic RNA viruses that have evolved a highly complex cascade of proteolytic processing events to control viral gene expression and replication. Most maturation cleavages within the precursor polyprotein are mediated by rhinovirus 3C protease (or its immediate precursor, 3CD), a cysteine protease with a trypsin-like polypeptide fold. High-resolution crystal structures of the enzyme from three viral serotypes have been used for the design and elaboration of 3C protease inhibitors representing different structural and chemical classes. Inhibitors having α,β-unsaturated carbonyl groups combined with peptidyl-binding elements specific for 3C protease undergo a Michael reaction mediated by nucleophilic addition of the enzyme’s catalytic Cys-147, resulting in covalent-bond formation and irreversible inactivation of the viral protease. Direct inhibition of 3C proteolytic activity in virally infected cells treated with these compounds can be inferred from dose-dependent accumulations of viral precursor polyproteins as determined by SDS/PAGE analysis of radiolabeled proteins. Cocrystal-structure-assisted optimization of 3C-protease-directed Michael acceptors has yielded molecules having extremely rapid in vitro inactivation of the viral protease, potent antiviral activity against multiple rhinovirus serotypes and low cellular toxicity. Recently, one compound in this series, AG7088, has entered clinical trials.

Genomic cloning of methylthioadenosine phosphorylase: a purine metabolic enzyme deficient in multiple different cancers.

5'-Deoxy-5'-methylthioadenosine phosphorylase (methylthioadeno-sine: ortho-phosphate methylthioribosyltransferase, EC 24.2.28; MTAP) plays a role in purine and polyamine metabolism and in the regulation of transmethylation reactions. MTAP is abundant in normal cells but is deficient in many cancers. Recently, the genes for the cyclin-dependent kinase inhibitors p16 and p15 have been localized to the short arm of human chromosome 9 at band p21, where MTAP and interferon alpha genes (IFNA) also map. Homozygous deletions of p16 and p15 are frequent malignant cell lines. However, the order of the MTAP, p16, p15, and IFNA genes on chromosome 9p is uncertain, and the molecular basis for MTAP deficiency in cancer is unknown. We have cloned the MTAP gene, and have constructed a topologic map of the 9p21 region using yeast artificial chromosome clones, pulse-field gel electrophoresis, and sequence-tagged-site PCR. The MTAP gene consists of eight exons and seven introns. Of 23 malignant cell lines deficient in MTAP protein, all but one had complete or partial deletions. Partial or total deletions of the MTAP gene were found in primary T-cell acute lymphoblastic leukemias (T-ALL). A deletion breakpoint of partial deletions found in cell lines and primary T-ALL was in intron 4. Starting from the centromeric end, the gene order on chromosome 9p2l is p15, p16, MTAP, IFNA, and interferon beta gene (IFNB). These results indicate that MTAP deficiency in cancer is primarily due to codeletion of the MTAP and p16 genes.

Diários fotográficos de bicicleta em Pernambuco: os irmãos Ulysses e Gilberto Freyre na documentação de cidades na década de 1920

Esta pesquisa interdisciplinar centrada no segmento da fotografia de acervo de intelectual apresenta o estudo de caso de 84 imagens feitas pelo fotógrafo amador Ulysses Freyre de alguns prédios e ruas das cidades de Olinda e do Recife entre 1923 e 1925. Ulysses fotografou durante passeios de bicicleta aos domingos ao lado do irmão, o sociólogo Gilberto Freyre. Objetiva-se traçar os dois usos dados por Gilberto às fotos de Ulysses: de base aos desenhos de Manoel Bandeira para o \"Livro do Nordeste\", organizado pelo sociólogo em 1925 para o centenário do Diário de Pernambuco; e como parte da concepção de inventário de edificações da arquitetura civil que serviu à Inspetoria de Monumentos Estaduais em 1928 em Pernambuco. Vale-se do campo acerca do circuito fotográfico nestas cidades, que estavam sob reformas urbanas no início do século XX, a fim de situar e revelar a fotografia de Ulysses como artefato de memória propulsor do embrionário projeto político-intelectual de Gilberto neste período. As fotos estão no acervo da Fundação Gilberto Freyre, em Recife, Pernambuco.

Reverse Geometry Hybrid Contact Lens Fitting in a Case of Donor-Host Misalignment after Keratoplasty

Purpose: To report the successful outcome obtained after fitting a new hybrid contact lens in a cornea with an area of donor-host misalignment and significant levels of irregular astigmatism after penetrating keratoplasty (PKP). Materials and methods: A 41-year-old female with bilateral asymmetric keratoconus underwent PKP in her left eye due to the advanced status of the disease. One year after surgery, the patient referred a poor visual acuity and quality in this eye. The fitting of different types of rigid gas permeable contact lenses was performed, but with an unsuccessful outcome due to contact lens stability problems and uncomfortable wear. Scheimpflug imaging evaluation revealed that a donor-host misalignment was present at the nasal area. Contact lens fitting with a reverse geometry hybrid contact lens (Clearkone, SynergEyes Carlsbad) was then fitted. Visual, refractive, and ocular aberrometric outcomes were evaluated during a 1-year period after the fitting. Results: Uncorrected distance visual acuity improved from a prefitting value of 20/200 to a best corrected postfitting value of 20/20. Prefitting manifest refraction was +5.00 sphere and -5.50 cylinder at 75°, with a corrected distance visual acuity of 20/30. Higher order root mean square (RMS) for a 5 mm pupil changed from a prefitting value of 6.83 µm to a postfitting value of 1.57 µm (5 mm pupil). The contact lens wearing was referred as comfortable, with no anterior segment alterations. Conclusion: The SynergEyes Clearkone contact lens seems to be another potentially useful option for the visual rehabilitation after PKP, especially in cases of donor-host misalignment.

Fitting of a New Design of Full Scleral Contact Lens in Advanced Keratoconus with Previous Implantation of Intracorneal Ring Segments

This case report reports the visual rehabilitation obtained with the fitting of a new design of full scleral contact lens (ICD 16.5 contact lens, Paragon Vision Sciences, distributed by Lenticon, Madrid, Spain) in a cornea with advanced keratoconus and previous implantation of intracorneal ring segment with a very limited effect. This eye had a refraction of –3.00 × 55° cylinder, providing a visual acuity of 0.5 LogMAR. The topographic pattern was very irregular with the presence of a significant central protrusion and a significant central corneal thinning. Some previous unsuccessful fittings have been performed with corneal and corneal-scleral lenses. A comfortable wearing was achieved with a fully scleral contact lens of 4600 μm of sagittal height, optical power of –11.25 D, and providing an apical clearance of 196 μm. A visual acuity of 0.0 LogMAR combined with a relevant aberrometric improvement was achieved with this contact lens. The patient was completely satisfied with the fitting. The result was maintained during 1 year after the fitting. Full scleral lenses are then able to provide comfortable wear and a significant increase in visual acuity combined with a significant improvement in the visual quality in eyes with advanced keratoconus.

Errors Associated to Keratoconus Grading using Systems based on Corneal Power

Purpose: To analyze and define the possible errors that may be introduced in keratoconus classification when the keratometric corneal power is used in such classification. Materials and methods: Retrospective study including a total of 44 keratoconus eyes. A comprehensive ophthalmologic examination was performed in all cases, which included a corneal analysis with the Pentacam system (Oculus). Classical keratometric corneal power (Pk), Gaussian corneal power (Pc Gauss), True Net Power (TNP) (Gaussian power neglecting the corneal thickness effect), and an adjusted keratometric corneal power (Pkadj) (keratometric power considering a variable keratometric index) were calculated. All cases included in the study were classified according to five different classification systems: Alió-Shabayek, Amsler-Krumeich, Rabinowitz-McDonnell, collaborative longitudinal evaluation of keratoconus (CLEK), and McMahon. Results: When Pk and Pkadj were compared, differences in the type of grading of keratoconus cases was found in 13.6% of eyes when the Alió-Shabayek or the Amsler-Krumeich systems were used. Likewise, grading differences were observed in 22.7% of eyes with the Rabinowitz-McDonnell and McMahon classification systems and in 31.8% of eyes with the CLEK classification system. All reclassified cases using Pkadj were done in a less severe stage, indicating that the use of Pk may lead to the classification of a cornea as keratoconus, being normal. In general, the results obtained using Pkadj, Pc Gauss or the TNP were equivalent. Differences between Pkadj and Pc Gauss were within ± 0.7D. Conclusion: The use of classical keratometric corneal power may lead to incorrect grading of the severity of keratoconus, with a trend to a more severe grading.

Non-alcoholic fatty liver disease associated with hypobetalipoproteinemia: report of three cases and a novel mutation in APOB gene

Background: Non-alcoholic fatty liver disease, the leading cause of chronic liver disease in children, is defined by hepatic fat infiltration >5% of hepatocytes, in the absence of excessive alcohol intake, evidence of viral, autoimmune or drug-induced liver disease. Conditions like rare genetic disorders must be considered in the differential diagnosis. Case Report: Two male brothers, and a non-related girl, all overweight, had liver steatosis. One of the brothers and the girl had elevated transaminases; all three presented with low total cholesterol, low density lipoproteins and very low density lipoproteins cholesterol levels, hypotriglyceridemia and low apolipoprotein B. A liver biopsy performed in the brother with citolysis confirmed steatohepatitis and the molecular study of apolipoprotein B gene showed a novel homozygous mutation (c.9353dup p.Asn3118Lysfs17). Patients with cytolysis lost weight, however liver steatosis persists. Conclusion: Fatty liver disease might be a consequence of hypobetalipoproteinemia. Evidence is scarce due to low number of reported cases.

Give me a shilling [videorecording]: phenomenon of the street children

Pictures of the Watoto wa Lwanga project, the efforts of the Brothers of St. Charles Lwanga not only to rehabilitate Nairobi's street children, but to prevent slum children from ending up in the streets. Features life in the slums, the situation of the street children, the efforts of the social workers to involve the children's parents, the various reception centers, the vocational school, the Boy's Town Ruai Residential School, the community of the Brothers of St. Charles Lwanga as well as interviews with former street children and with various directors and benefactors of the project.

Letter of recommendation for Elisha Parmele and Reuben Parmele, May 17, 1784

This letter, signed by Stephen West, Ezra Stiles, and Josiah Sherman, attests to the reputable character of Elisha Parmele and his brother, Reuben, who was also a minister. The signers explain the health concerns motivating the brothers' travels southward and request that those who encounter them treat them with "kindness and brotherly love."