RESUMO
The Na+-Cl- cotransporter (NCC; SLC12A3) is a highly regulated integral membrane protein that is known to exist as three splice variants in primates. Its primary role in the kidney is to mediate the cosymport of Na+ and Cl- across the apical membrane of the distal convoluted tubule. Through this role and the involvement of other ion transport systems, NCC allows the systemic circulation to reclaim a fraction of the ultrafiltered Na+, K+, Cl-, and Mg+ loads in exchange for Ca2+ and [Formula: see text]. The physiological relevance of the Na+-Cl- cotransport mechanism in humans is illustrated by several abnormalities that result from NCC inactivation through the administration of thiazides or in the setting of hereditary disorders. The purpose of the present review is to discuss the molecular mechanisms and overall roles of Na+-Cl- cotransport as the main topics of interest. On reading the narrative proposed, one will realize that the knowledge gained in regard to these themes will continue to progress unrelentingly no matter how refined it has now become.
Assuntos
Homeostase , Humanos , Animais , Homeostase/fisiologia , Membro 3 da Família 12 de Carreador de Soluto/metabolismo , Equilíbrio Hidroeletrolítico/fisiologia , Sódio/metabolismo , Rim/metabolismoRESUMO
PURPOSE: Cryofibrinogenemia is a rare cryopathy presenting as acrocyanosis following exposure to cold. Familial presentation has been described but the underlying molecular cause remained undetermined. METHODS: Forty (40) members from a large family with an initial diagnosis of familial cryofibrinogenemia were interviewed and examined to determine affected status and collect DNA. Exome sequencing was performed on three affected individuals from distinct branches of the pedigree. RESULTS: Seventeen (17) family members reported a history of acrocyanosis with cold exposure. None reported symptoms were suggestive of lupus. Exome sequencing of three subjects identified the heterozygous mutation D18N in the TREX1 gene which was then confirmed by Sanger sequencing in all affected as well as 2 unaffected family members. The mutation is already being associated with familial chilblain lupus erythematosus (CHLE), and a systematic review of literature was undertaken to compare reports of familial CHLE and cryofibrinogenemia. Both entities were found to share highly similar clinical presentations suggesting they are part of a same syndrome in which cryofibrinogenemia and lupus manifestations have variable penetrance. CONCLUSIONS: Familial cryofibrinogenemia without lupus should be added to the spectrum of TREX1-related disease.
Assuntos
Crioglobulinemia/genética , Exodesoxirribonucleases/genética , Fosfoproteínas/genética , Adulto , Pérnio/genética , DNA/genética , Feminino , Predisposição Genética para Doença/genética , Heterozigoto , Humanos , Lúpus Eritematoso Cutâneo/genética , Lúpus Eritematoso Sistêmico/genética , Masculino , Mutação/genética , LinhagemRESUMO
Familial focal epilepsy with variable foci (FFEVF) is a heterogeneous epilepsy syndrome originally described in the French-Canadian (FC) population. Mutations in DEPDC5 have recently been identified in multiple cases of FFEVF as well as in a wide spectrum of other familial focal epilepsies. In this study, we aimed to determine the frequency of mutation of this gene in our large cohort of FC individuals with FFEVF, as well as familial and sporadic cases with focal epilepsy. We report a recurrent p.R843X protein-truncating mutation segregating in one large FFEVF and two small focal epilepsy FC families. Fine genotyping suggests an ancestral allele. A new p.T864M variant, predicted to be disease-causing, was also identified in a small FC family. Overall, we identified DEPDC5 mutations in 5% of our familial and sporadic focal epilepsy cases (4/79). Our results support the view that mutations in the DEPDC5 gene are an important cause of autosomal dominant focal epilepsies in the FC population, including a founder mutation that is specific to this population. These findings may facilitate molecular diagnosis in clinical practice.
Assuntos
Epilepsias Parciais/genética , Predisposição Genética para Doença , Mutação , Proteínas Repressoras/genética , Adolescente , Adulto , Canadá/etnologia , Criança , Pré-Escolar , Feminino , Proteínas Ativadoras de GTPase , Humanos , Masculino , LinhagemRESUMO
Predation is considered one of the main costs to reproduction but is rarely examined from a physiological perspective. In particular, little is known about the influence of brood predation pressure on the physiology of parents engaged in care. Brood defense, even when there is no direct threat to the parent, can be costly as it requires constant vigilance and chasing predators to protect the developing brood and maintain parental investment (i.e., fitness). Our goal was to examine the influence of natural variation in nest predation pressure on the physiology of the teleost smallmouth bass Micropterus dolomieu, an animal that provides sole-paternal care for developing offspring. More specifically, we used indicators of anaerobic (lactate dehydrogenase [LDH]) and aerobic capacity (cytochrome c oxidase [CCO] and citrate synthase [CS]) in axial white muscle and pectoral red muscle to test for differences in antipredator performance of nest guarding males across six lakes with natural variation in nest predation pressure. Pectoral red muscle enzyme activities and protein concentrations were highly conserved among populations, while axial white muscle showed differences in LDH activities, CCO activities and protein concentrations. However, there was no evidence for higher metabolic capacities in fish from lakes with increased brood predation pressure. Clearly, factors other than predation pressure have a greater influence on white muscle metabolic capacities. Additional research is needed to clarify the extent to which biotic and abiotic factors influence the enzyme activity and organismal performance in wild animals, particularly at the level of the individual and population.
Assuntos
Peixes/fisiologia , Comportamento de Nidação/fisiologia , Comportamento Predatório/fisiologia , Estresse Psicológico/enzimologia , Natação/fisiologia , Aerobiose/fisiologia , Animais , Citrato (si)-Sintase/metabolismo , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Ativação Enzimática/fisiologia , Feminino , Peixes/metabolismo , L-Lactato Desidrogenase/metabolismo , Masculino , Músculo Esquelético/enzimologia , Músculo Esquelético/metabolismo , Proteínas/análise , Proteínas/metabolismo , Estresse Psicológico/metabolismoRESUMO
The energetic and physiological status of parental smallmouth bass Micropterus dolomieu was investigated across the majority of their latitudinal range at the onset and near the end of care. Variables such as tissue lipid stores, plasma indicators of nutritional status and chronic stress and white muscle were used to define energetic and physiological status. Results showed that northern males (48 degrees N) were larger and heavier than mid-northern (44 degrees N) and southern (36 degrees N) latitude males. For a given body size, northern males had greater whole-body lipid across the parental care period and tended to feed more (based on gut contents) than mid-northern and southern latitude conspecifics. Indicators of nutritional status were also highest in northern males. Conversely, the southern males exhibited the greatest capacity for biosynthesis across the entire parental care period as indicated by the highest level of nucleoside diphosphate kinase activities. Collectively, these finding suggest that the energetic costs and physiological consequences of care vary across latitudes, providing some of the first mechanistic evidence of how environmental conditions can influence both the ecological and physiological costs of reproduction for wild animals during parental care. The data also suggest that lake-specific processes that can vary independently of latitude may be important, necessitating additional research on fish reproductive physiology across landscapes.
Assuntos
Bass/fisiologia , Comportamento Animal/fisiologia , Metabolismo Energético/fisiologia , Animais , Proteínas Sanguíneas/análise , Colesterol/sangue , Enzimas/metabolismo , Conteúdo Gastrointestinal/química , Masculino , Sódio/sangue , Triglicerídeos/sangueRESUMO
Previous studies have suggested the persistence of oligodendrocyte progenitor cells in the adult mammalian subcortical white matter. To identify oligodendrocyte progenitors in the adult human subcortical white matter, we transfected dissociates of capsular white matter with plasmid DNA bearing the gene for green fluorescence protein (hGFP), placed under the control of the human early promoter (P2) for the oligodendrocytic protein cyclic nucleotide phosphodiesterase (P/hCNP2). Within 4 d after transfection with P/hCNP2:hGFP, a discrete population of small, bipolar cells were noted to express GFP. These cells were A2B5-positive (A2B5(+)), incorporated bromodeoxyuridine in vitro, and constituted <0.5% of all cells. Using fluorescence-activated cell sorting (FACS), the P/hCNP2-driven GFP(+) cells were then isolated and enriched to near-purity. In the weeks after FACS, most P/hCNP2:hGFP-sorted cells matured as morphologically and antigenically characteristic oligodendrocytes. Thus, the human subcortical white matter harbors mitotically competent progenitor cells, which give rise primarily to oligodendrocytes in vitro. By using fluorescent transgenes of GFP expressed under the control of an early oligodendrocytic promoter, these oligodendrocyte progenitor cells may be extracted and purified from adult human white matter in sufficient numbers for implantation and cell-based therapy.
Assuntos
2',3'-Nucleotídeo Cíclico Fosfodiesterases/genética , Oligodendroglia/citologia , Regiões Promotoras Genéticas , Prosencéfalo/citologia , Células-Tronco/citologia , Adulto , Separação Celular/métodos , Citometria de Fluxo/métodos , Genes Reporter , Proteínas de Fluorescência Verde , Humanos , Proteínas Luminescentes/análise , Proteínas Luminescentes/genética , Proteínas Recombinantes/análise , Transfecção/métodosRESUMO
We demonstrated previously that platelet-activating factor (PAF), a potent inflammatory mediator, acts on osteoclasts to elevate cytosolic [Ca2+] and stimulate resorption. However, it is not clear whether the effects of PAF on resorptive activity are direct or indirect. In the present study, we investigated the effects of PAF on osteoclast motility. Osteoclasts were isolated from the long bones of neonatal rabbits, and cell motility and morphology were monitored using time-lapse video microscopy. Calcitonin, a hormone known to induce retraction of pseudopods and inhibit resorptive activity, was used to render osteoclasts quiescent. Within 10 minutes of calcitonin treatment (100 ng/ml, final), pronounced retraction of pseudopods was observed in 68 of 112 cells tested. When PAF (200 nM, final) was added 10 minutes after calcitonin treatment, pseudopods were evident 1 h later in 15 of 37 calcitonin-responsive cells tested. In contrast, pseudopods were evident in only 4 of 31 calcitonin-responsive cells treated with control solutions (PAF-vehicle or S-PAF, the biologically inactive stereoisomer of PAF). Pseudopod formation was quantified by measuring the planar area of pseudopods with a computer-based video analysis system. When assessed 60 minutes following PAF treatment, the pseudopod area was significantly greater in PAF-treated cells than in control cells. In some calcitonin-treated osteoclasts, PAF induced pseudopod formation when applied focally using an extracellular micropipette, consistent with a direct action of PAF. We conclude that PAF directly induces pseudopod formation in calcitonin-inhibited osteoclasts, a morphologic response indicative of osteoclast activation.
Assuntos
Calcitonina/farmacologia , Cálcio/metabolismo , Osteoclastos/efeitos dos fármacos , Fator de Ativação de Plaquetas/toxicidade , Animais , Animais Recém-Nascidos , Reabsorção Óssea/tratamento farmacológico , Calcitonina/uso terapêutico , Movimento Celular/efeitos dos fármacos , Distribuição de Qui-Quadrado , Processamento de Imagem Assistida por Computador , Osteoclastos/ultraestrutura , Coelhos , Estereoisomerismo , Gravação em VídeoRESUMO
OBJECTIVE: To describe sleep and body movement patterns in incontinent nursing home residents for the purpose of determining if the residents require nighttime changing and body repositioning on a 2-hour schedule. DESIGN: Cross-sectional survey. SETTING: Four nursing homes. PARTICIPANTS: 118 nursing home residents. MEASUREMENTS: Over two nights, bedside monitoring equipment recorded wrist activity (as a proxy measure for sleep) and body movements of both the shoulder and hip areas in consecutive 2-minute intervals. Specific outcome measures were: (1) Average duration of a sleep episode, peak duration of a sleep episode, and percent of time in bed asleep. (2) The number of 2-minute intervals in which a large movement (45 degree turn) at the shoulder and hip was noted per hour of recording for each resident. (3) The number of resident-initiated, rather than staff-initiated, large movements at the shoulder and hip that occurred within the same 2-minute intervals. RESULTS: There was large variability in all sleep measures; however, on average, residents slept 66% of the time they were in bed. The distribution of these measures suggests that sleep was punctuated with frequent nighttime awakenings. Thirty-three percent of the incontinent residents demonstrated very low levels of resident-initiated movement at the shoulder and hip. Sixty-six percent demonstrated at least one large movement at the shoulder and hip per hour during periods of sleep as well as during periods of wake. CONCLUSION: The majority of incontinent nursing home residents self-initiate sufficiently frequent movements at both the shoulder and hip so as not to be in need of frequent repositioning by nursing staff. Since the sleep of many of these residents is also characterized by frequent awakenings, incontinent nursing home residents may benefit from a schedule of nursing care at night that considers sleep of equal importance to incontinence care and body repositioning.
Assuntos
Instituição de Longa Permanência para Idosos , Casas de Saúde , Sono , Incontinência Urinária/fisiopatologia , Idoso , Idoso de 80 Anos ou mais , Estudos Transversais , Feminino , Humanos , Masculino , Movimento , Assistência Noturna/métodos , Polissonografia , Úlcera por Pressão/enfermagem , Úlcera por Pressão/fisiopatologia , Sensibilidade e Especificidade , Sono/fisiologia , Incontinência Urinária/enfermagemRESUMO
OBJECTIVE: To evaluate the association between noise, light, nursing care practices, and nighttime awakenings in incontinent nursing home residents. DESIGN: Cross-sectional survey. SETTING: Four long-term care nursing facilities. PARTICIPANTS: One hundred eighteen incontinent nursing home residents. MEASUREMENTS: Over two nights, bedside monitoring equipment recorded wrist activity, resident bed movements, and environmental noise and light changes in consecutive 2-minute intervals. Changes in sleep and bed movement were compared with changes in noise and light that occurred within the same or proximal 2-minute intervals. Noise and light changes in combination with large resident movement at the hip and shoulder were interpreted as related to incontinence care based on observational measures. Specific outcome measures were: (1) the number of noise and light changes as well as staff care practices that did not wake the resident during periods of consecutive sleep, ie, sleep lasting a minimum of 10 minutes; (2) the number of noise and light changes as well as staff care practices that occurred immediately before or during the 2-minute intervals during which a resident woke from a period of consecutive sleep; and (3) the number of such staff care practices that were related to incontinence care. RESULTS: Noise and light changes associated with both general environmental events and more specific nurse care practices were associated with 50% of all waking episodes of 4 minutes or longer and 35% of all waking episodes of 2 minutes or shorter. The major sources of all noise were traced to nursing staff. Eighty-seven percent of all incontinence care practices were associated with episodes of waking. CONCLUSION: The data reported in this paper document that general environmental noise and incidents of nursing care practices, particularly those related to incontinence care, are responsible for a substantial amount of the sleep fragmentation that is common among nursing home residents.
Assuntos
Ambiente de Instituições de Saúde , Instituição de Longa Permanência para Idosos , Assistência Noturna , Casas de Saúde , Privação do Sono , Incontinência Urinária/enfermagem , Vigília , Idoso , Idoso de 80 Anos ou mais , Estudos Transversais , Feminino , Quadril/fisiologia , Humanos , Iluminação/efeitos adversos , Masculino , Atividade Motora/fisiologia , Assistência Noturna/métodos , Ruído/efeitos adversos , Pesquisa em Avaliação de Enfermagem , Recursos Humanos de Enfermagem , Polissonografia , Padrões de Prática Médica , Ombro/fisiologia , Privação do Sono/fisiologia , Fatores de Tempo , Vigília/fisiologia , Punho/fisiologiaRESUMO
We report the clinical course and pathologic findings in a case of intraocular sclerosing inflammatory pseudotumor in a 21-year-old man. The patient initially had a unilateral right interstitial keratitis, scleritis, uveitis, ciliary body mass, and retinal detachment. Scleral and vitreous biopsy specimens revealed an inflammatory process. The eye was eventually enucleated despite therapy with high doses of prednisone and ciprofloxacin hydrochloride. Histologic examination of the globe showed nongranulomatous, acute (neutrophils) and chronic (lymphocytes and histiocytes) inflammation with proliferation of fibrous tissue within the vitreous cavity, uvea, sclera, and contiguous orbital fibroadipose tissue. The contralateral eye later developed a similar mass that resolved following aggressive and prolonged immunosuppressive therapy with retention of 20/16 visual acuity.
Assuntos
Oftalmopatias/patologia , Granuloma de Células Plasmáticas/patologia , Esclera/patologia , Corpo Vítreo/patologia , Adulto , Oftalmopatias/terapia , Enucleação Ocular , Granuloma de Células Plasmáticas/terapia , Histiócitos/patologia , Humanos , Imunossupressores/uso terapêutico , Ceratite/patologia , Linfócitos/patologia , Masculino , Neutrófilos/patologia , Descolamento Retiniano/patologia , Esclerite/patologia , Esclerose/patologia , Uveíte/patologia , Acuidade VisualRESUMO
Previous research has shown that glucose can enhance memory in animals and humans. In humans, the facilitative effect of glucose is best observed with declarative memory tasks in older subjects. While the memory-enhancing action of glucose is well established, the underlying physiological mechanisms and the specific aspects of memory that are modulated by glucose in humans are not well understood. The present study sought to examine the effects of glucose on memory in young women using a memory paradigm sensitive to specific encoding and retrieval strategies. The glucose dose was adjusted for the weight of each participant in order to generate a dose response curve covering most doses used in previous studies. The results showed that 300 mg/kg glucose enhanced the primacy effect as defined by the recall of the first five items of the lists. However, none of the doses of glucose produced changes in the recall priority given to primacy items. The effect of glucose appears to be localized on the recall primacy effect, suggesting that glucose acts on precise memory operations. This improvement, however, is independent of the order in which subjects recalled the words. Cholinergic drugs have been shown to alter the recall of the primacy part of word lists and this observation is consistent with the hypothesis that glucose acts on memory through an interaction with brain cholinergic systems.
Assuntos
Cognição/efeitos dos fármacos , Glucose/administração & dosagem , Rememoração Mental/efeitos dos fármacos , Acetilcolina/metabolismo , Adolescente , Adulto , Glicemia , Encéfalo/metabolismo , Feminino , Humanos , Pessoa de Meia-Idade , Desempenho Psicomotor/fisiologiaRESUMO
The use of air biofiltration for the degradation of dichlorobenzenes (1,2-DCB and 1,4-DCB) was studied at a refinery site. At this plant, 93 m3/h of contaminated groundwater, used in a cooling system and containing a maximum of 2 ppm of dichlorobenzenes, had to be treated. Stripping of the DCBs followed by biofiltration was selected as the most suitable technology to avoid volatilization in ambient air as expected with a wastewater aerobic treatment system. A stripper of 15 m height and 1.27 m diameter was designed as a first step treatment to volatilize DCBs with 3400 m3/h of air. Two f ull-scale biofilters of 70 m3 each were built and filled with 45 m3 of filtering media for the adsorption and biodegradation of the DCBs in the gas-phase. Filtering media was composed mainly of peat moss, with animal manure, wood chips and DCBs contaminated soil. Air to be treated was also contaminated with naphthalene. Laboratory tests showed an effective microbial activity in the contaminated soil and in the filtering media for DCBs degradation. Degradation of naphthalene induced slower degradation of DCBs. Full-scale operation was studied during four months. Water flow and DCBs content in the water entering the stripper were lower than expected with only 57 m3/h and a maximum concentration of only 240 ppb. Effective desorption was obtained in the stripper in the full-scale operation (more than 99% removal). Full-scale biofilters maintained a DCB concentration of less than 1 ppmv in the air outlet, but removal efficiency varied between 0 and 79% because of the low DCB inlet concentrations, load variations and sporadic naphthalene presence.
Assuntos
Derivados de Benzeno/metabolismo , Compostos Clorados/metabolismo , Poluentes Ambientais/metabolismo , Adsorção , Bactérias Aeróbias/fisiologia , Biodegradação Ambiental , Filtração , Esterco , Eliminação de Resíduos , Volatilização , Movimentos da ÁguaRESUMO
The key to the efficiency of N,N-diethanolaminomethyl polystyrene (DEAM-PS), the first solid support capable of coupling to boronic acids, is the formation of a stable, resin-bound boronic ester ate adduct (see scheme). With this resin it is now possible to efficiently immobilize a wide variety of boronic acids including functionalized ones that can be derivatized by solid-phase combinatorial synthesis.
RESUMO
Yersinia enterocolitica serotype 0:3, biotype 4, has been isolated from two family members with diarrhea and from the well used as a source of their drinking water.
Assuntos
Microbiologia da Água , Abastecimento de Água , Yersiniose/genética , Yersinia enterocolitica/isolamento & purificação , Adulto , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Yersiniose/etiologiaRESUMO
Although transplantation centers directly benefit from organ and tissue donation, they continue to yield low organ and tissue referral and donation rates. Our medical center and organ procurement organization developed a model to increase referral and donation rates. This model, called the Transplant Center Development Model, facilitates the donation process, specializes staff education, and promotes administrative involvement. After it was was implemented at our medical center in 1991, the referral and donation rates from 1988 to 1990 were compared with those from 1991 to 1993. The results showed that after implementation of the model, the organ referral mean increased 47%; the organ donation mean, 50%; and the tissue donation mean, 117%. These findings suggest that this model may be a valuable tool in transplant center development.
Assuntos
Modelos Organizacionais , Encaminhamento e Consulta , Obtenção de Tecidos e Órgãos/organização & administração , Pessoal de Saúde/educação , Humanos , Avaliação de Programas e Projetos de Saúde , Estudos Retrospectivos , Doadores de TecidosRESUMO
Plasmid pPM114 carries the Escherichia coli 16S ribosomal RNA gene under the control of a T7 promoter. It can generate in vitro transcribed 16S rRNA that can be assembled into functional 30S ribosomal subunits. Two deletion mutants were derived from pPM114, by partial or total deletion of the conserved 900 stem/loop region of the 16S rRNA. These mutants, pMG delta 10 and pMG delta 23, respectively lack bases 895 to 904 and 889 to 911 of the 16S rRNA. The amputated 16S rRNA transcripts synthesized from these mutated plasmids were assembled into 30S subunits which were as active under the direction of an artificial or a natural messenger as subunits reconstructed with the full-length 16S rRNA transcript. They also responded as well to the stimulation of misreading by streptomycin, although the deleted region is proximal to the streptomycin binding domain. However, when we attempted to delete the 895-904 or 889-911 region from the 16S rRNA gene in plasmid pKK3535 which carries the rrnB operon, no transformants harbouring plasmids with one of these deletions could be recovered. These observations suggest that the 900 stem/loop region of the 16S rRNA is not required for the ribosomal function but is probably essential for important cell regulatory functions.
Assuntos
Proteínas de Bactérias/biossíntese , Escherichia coli/genética , RNA Ribossômico 16S/fisiologia , RNA Ribossômico/fisiologia , Proteínas Ribossômicas/biossíntese , Sequência de Bases , Deleção Cromossômica , Resistência Microbiana a Medicamentos/genética , Escherichia coli/efeitos dos fármacos , Dados de Sequência Molecular , RNA Ribossômico 16S/efeitos dos fármacos , Estreptomicina/farmacologia , Transcrição Gênica/efeitos dos fármacosRESUMO
Cadherins form a large family of membrane glycoproteins which mediate homophilic calcium-dependent cell adhesion. They are thought to mediate the initial calcium-dependent cell adhesion which precedes the plasma membrane fusion of skeletal myoblasts. Two cadherin subtypes are known to be expressed in mammalian skeletal myoblasts: muscle cadherin (M-cadherin) and neural cadherin (N-cadherin). In the present study we demonstrate that 1) the expression of M- and N-cadherin is differentially regulated during myoblast differentiation in vitro, 2) the expression of M-cadherin but not N-cadherin is inhibited by 5-bromo-2'-deoxyuridine (BUdR), an agent which selectively inhibits skeletal myoblast differentiation, and 3) fusion and differentiation-competent rat L6 myoblasts do not express detectable levels of N-cadherin mRNA. In vivo, M-cadherin mRNA was detectable exclusively in skeletal muscle. M-cadherin mRNA levels peaked during the secondary myogenic wave in rat hindlimb muscle, becoming barely detectable in 1-week-old and adult rats. These observations indicate that M-cadherin is unique in two ways: It is the first cadherin to be included in the family of skeletal muscle-specific genes, and it shows peak levels of expression in developing skeletal muscle tissue. Taken together, these results suggest that M-cadherin plays an important role in skeletal myogenesis.
Assuntos
Envelhecimento/metabolismo , Animais Recém-Nascidos/metabolismo , Caderinas/metabolismo , Músculo Esquelético/citologia , Animais , Animais Recém-Nascidos/crescimento & desenvolvimento , Sequência de Bases , Bromodesoxiuridina/farmacologia , Caderinas/genética , Diferenciação Celular , Linhagem Celular , Camundongos , Dados de Sequência Molecular , Desenvolvimento Muscular , Músculo Esquelético/crescimento & desenvolvimento , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-DawleyRESUMO
We have isolated a cDNA coding for the larger isoform of the rat brain 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNP2), a protein associated with myelination in the central nervous system (CNS). The complete 420 amino acid sequence was deduced from the nucleotide sequence of the cDNA. Sequence comparisons show that rat CNP shares 96% homology with mouse, 84% with bovine, and 86% with human CNP. Errors in the published sequence of rat CNP1 have now been corrected. Comparisons with other proteins reveal several interesting conserved motifs, including two leucine repeat heptads, and two consensus motifs for phosphorylation in the N-terminal domain of CNP2.
Assuntos
3',5'-AMP Cíclico Fosfodiesterases/metabolismo , Encéfalo/enzimologia , Isoenzimas/metabolismo , 3',5'-AMP Cíclico Fosfodiesterases/biossíntese , Sequência de Aminoácidos , Animais , Sequência de Bases , Northern Blotting , Bovinos , Clonagem Molecular , DNA/análise , Humanos , Isoenzimas/biossíntese , Camundongos , Dados de Sequência Molecular , Ratos , Homologia de Sequência do Ácido NucleicoRESUMO
A plasmid has been constructed, which contains the 16S ribosomal RNA gene of Escherichia coli immediately downstream from a phage T7 promoter. In vitro transcription of this gene by RNA polymerase of the T7 phage yielded an unmethylated 16S rRNA that could be used for the assembly of functional 30S subunits. These subunits when assayed in a poly(U)-directed translation assay, were as active as 30S subunits reconstructed with native 16S rRNA. This system opens the possibility of investigating the role of the methylations of the rRNA and the functional consequences of site-directed mutagenesis in a rRNA gene. An application of this system was provided by generating a 16S rRNA transcript shortened by about 30 nucleotides at the 3' end. This truncated rRNA could be reassembled into 30S subunits, about 70% as active as 30S subunits reconstructed with the full-length 16S rRNA transcript.