RESUMO
The specificity and implementation of current MRI-based diagnostic criteria for multiple sclerosis (MS) are imperfect. Approximately 1 in 5 of individuals diagnosed with MS are eventually determined not to have the disease, with overreliance on MRI findings a major cause of MS misdiagnosis. The central vein sign (CVS), a proposed MRI biomarker for MS lesions, has been extensively studied in numerous cross sectional studies and may increase diagnostic specificity for MS. CVS has desirable analytical, measurement, and scalability properties. "Central Vein Sign: A Diagnostic Biomarker in Multiple Sclerosis (CAVS-MS)" is an NIH-supported, 2-year, prospective, international, multicenter study conducted by the North American Imaging in MS Cooperative (NAIMS) to evaluate CVS as a diagnostic biomarker for immediate translation into clinical care. Study objectives include determining the concordance of CVS and McDonald Criteria to diagnose MS, the sensitivity of CVS to detect MS in those with typical presentations, and the specificity of CVS among those with atypical presentations. The study will recruit a total of 400 participants (200 with typical and 200 with atypical presentations) across 11 sites. T2*-weighted, high-isotropic-resolution, segmented echo-planar MRI will be acquired at baseline and 24 months on 3-tesla scanners, and FLAIR* images (combination of FLAIR and T2*) will be generated for evaluating CVS. Data will be processed on a cloud-based platform that contains clinical and CVS rating modules. Imaging quality control will be conducted by automated methods and neuroradiologist review. CVS will be determined by Select6* and Select3* lesion methods following published criteria at each site and by central readers, including neurologists and neuroradiologists. Automated CVS detection and algorithms for incorporation of CVS into McDonald Criteria will be tested. Diagnosis will be adjudicated by three neurologists who served on the 2017 International Panel on the Diagnosis of MS. The CAVS-MS study aims to definitively establish CVS as a diagnostic biomarker that can be applied broadly to individuals presenting for evaluation of the diagnosis of MS.
Assuntos
Esclerose Múltipla , Biomarcadores , Estudos Transversais , Humanos , Imageamento por Ressonância Magnética , Estudos Multicêntricos como Assunto , Esclerose Múltipla/diagnóstico por imagem , Estudos ProspectivosRESUMO
Atopic dermatitis (AD) is a common immune-medicated skin disease. Previous studies have explored the relationship between Human Leukocyte Antigen (HLA) allelic variation and AD with conflicting results. The aim was to examine HLA Class I genetic variation, specifically peptide binding groove variation, and associations with AD. A case-control study was designed to evaluate HLA class I allelic variation and binding pocket polymorphisms, using next generation sequencing on 464 subjects with AD and 388 without AD. Logistic regression was used to evaluate associations with AD by estimating odds ratios (95% confidence intervals). Significant associations were noted with susceptibility to AD (B*53:01) and protection from AD (A*01:01, A*02:01, B*07:02 and C*07:02). Evaluation of polymorphic residues in Class I binding pockets revealed six amino acid residues conferring protection against AD: A9F (HLA-A, position 9, phenylalanine) [pocket B/C], A97I [pocket C/E], A152V [pocket E], A156R [pocket D/E], B163E [pocket A] and C116S [pocket F]. These findings demonstrate that specific HLA class I components are associated with susceptibility or protection from AD. Individual amino acid residues are relevant to protection from AD and set the foundation for evaluating potential HLA Class I molecules in complex with peptides/antigens that may initiate or interfere with T-cell responses.
Assuntos
Dermatite Atópica/genética , Predisposição Genética para Doença , Variação Genética , Antígenos de Histocompatibilidade Classe I/genética , Alelos , Estudos de Casos e Controles , Dermatite Atópica/diagnóstico , Frequência do Gene , Estudos de Associação Genética , Genótipo , Antígenos de Histocompatibilidade Classe I/química , Humanos , Modelos Moleculares , Razão de Chances , Polimorfismo de Nucleotídeo Único , Conformação Proteica , Análise de Sequência de DNA , Relação Estrutura-AtividadeRESUMO
INTRODUCTION: Sepsis is the leading cause of infectious morbidity and mortality among hospitalized neonates. In high-resource pediatric and adult intensive care units, use of aqueous chlorhexidine (CHG) solution has been associated with reduced risk of bloodstream infections (BSI). OBJECTIVES: To assess the impact of bathing of neonates with 2% CHG on BSI, suspected sepsis, and mortality in a low-income country neonatal care unit. METHODS: We conducted a secondary analysis of data from the Sepsis Prevention in Neonates in Zambia (SPINZ) study, a prospective observational cohort study performed at a large public referral hospital in Lusaka, Zambia. The SPINZ study assessed the impact of an infection control bundle (consisting of alcohol hand rub, SMS hygiene reminders, enhanced environmental cleaning, and CHG baths for babies ≥1.5 kg) on sepsis, BSI, and all-cause mortality. Episodic shortages in study staffing resulted in some enrolled babies not receiving a CHG bath. Using Longitudinal Targeted Maximum Likelihood Estimation and Cox proportional hazards regression to adjust for observed confounding, we estimated the causal effect of receiving a CHG bath within the first 3 days of life on suspected sepsis, BSI, and death among inborn babies enrolled during the study implementation and intervention phases. RESULTS: The majority of inborn, enrolled babies ≥1.5 kg received a CHG bath within 3 days of NICU admission (864 of 1233, 70%). We found that CHG bathing reduced the hazard rate of BSI among inborn babies ≥1.5 kg by a factor of 0.58 (p = 0.10, 95% CI: 0.31, 1.11), corresponding to an absolute risk reduction of 9.6 percentage points within a week of admission (p = 0.002, 95% CI: 3.4-15.7 percentage points). We did not find a statistically significant effect of CHG bathing on culture-negative sepsis (p = 0.54) or death (p = 0.85). CONCLUSION: In our single center study, CHG bathing at admission was associated with a reduced risk of BSI due to a pathogenic organism after adjusting for potential confounding. Our results suggest that CHG may be an effective intervention for preventing neonatal sepsis in high-risk, low-income country settings.
Assuntos
Clorexidina , Controle de Infecções , Sepse/prevenção & controle , Banhos , Estudos de Coortes , Feminino , Mortalidade Hospitalar , Humanos , Higiene , Recém-Nascido , Unidades de Terapia Intensiva Neonatal , Masculino , Estudos Prospectivos , ZâmbiaRESUMO
Naphthoquinones have been investigated as potential therapeutic molecules for neurodegenerative disorders, which is largely based on their anti-oxidative potential. However, a theoretical framework for the pleiotropic protective effects of naphthoquinone derivatives is largely missing. We synthesized a library of novel short chain 2,3-disubstituted naphthoquinone derivatives and measured their redox characteristics to identify a potential connection with their biological activity. Using two cell lines with different reducing potential, the compounds were tested for their inherent toxicity, acute rescue of ATP levels and cytoprotective activity. For the first time, a structure-activity-relationship for naphthoquinones has been established. Our results clearly demonstrate that it is the group on the alkyl side chain and not solely the redox characteristics of the naphthoquinone unit or lipophilicity that determines the extent of cytoprotection by individual compounds. From this, we developed a number of amide containing naphthoquinones with superior activity in ATP rescue and cell viability models compared to the clinically used benzoquinone idebenone.
RESUMO
Chlorpyrifos (CPF), an organophosphate pesticide inhibits acetylcholinesterase (AChE) and causes neuromuscular incoordination among children and elderly. The objectives of the present study were to compare the neurotoxic effects of dermal application of CPF on the cerebellum in the parameters of glial fibrillary acidic protein (GFAP) expression in young and adult mice and to correlate with the changes in acetylcholinesterase levels. Male Balb/c mice, 150 days old (adult) and 18 days old (young) were dermally applied with ½ LD(50) of CPF over the tails for 14 days. Serum AChE concentration was estimated and GFAP immunostaining was performed on sagittal paraffin sections through the vermis of cerebellum. Although reduced in both age-groups exposed to CPF, percentage of reduction in serum AChE was more in adult compared to the young. Under GFAP immunostaining, brown colour fibres and glial cells were observed in cerebellar cortex and medulla in both the experimental groups. The mean GFAP-positive glial cell count in cerebellar medulla per mm(2) of section was significantly (p < 0.05) increased in adult mice exposed to CPF when compared with age-matched control. In conclusion, this study confirmed that dermal exposure of CPF was able to exert neurotoxic effect in both young and adult mice. However, the quantitative results revealed that adult mice showed more GFAP expression in cerebellum when compared with the young, when exposed to CPF.
Assuntos
Cerebelo/efeitos dos fármacos , Clorpirifos/toxicidade , Inibidores da Colinesterase/toxicidade , Inseticidas/toxicidade , Acetilcolinesterase/sangue , Administração Cutânea , Fatores Etários , Animais , Astrócitos/efeitos dos fármacos , Astrócitos/metabolismo , Cerebelo/metabolismo , Clorpirifos/administração & dosagem , Inibidores da Colinesterase/administração & dosagem , Proteína Glial Fibrilar Ácida/metabolismo , Inseticidas/administração & dosagem , Masculino , Camundongos , Camundongos Endogâmicos BALB CRESUMO
The alteration of craniofacial structures has been associated with obstructive sleep apnoea (OSA). We hypothesised that: 1) a smaller mandible is a risk factor for OSA; and 2) the previously observed inferiorly positioned hyoid bone in apnoeics is associated with enlarged tongue volume. This is a case-control study using three-dimensional magnetic resonance imaging cephalometry. 55 apneics and 55 controls were matched for age, sex and race. The analysis was stratified by sex and controlled for age, race, height, neck visceral fat, skeletal type and tongue volume. We found that a 1-sd increase in mandibular length and depth were associated with decreased risk of sleep apnoea (OR 0.52, 95% CI 0.28-0.99 and OR 0.46, 95% CI 0.23-0.91, respectively) in males but not in females. Greater hyoid-to-nasion (OR 2.64, 95% CI 1.19-5.89 in males and OR 5.01, 95% CI 2.00-12.52 in females) and supramentale-to-hyoid (OR 2.39, 95% CI 1.12-5.14) in males and OR 3.38, 95% CI 1.49-7.68 in females) distances were associated with increased risk of OSA. The difference for hyoid position between apnoeics and controls was lost after controlling for tongue volume. Enlargement of tongue is likely to be the pathogenic factor for inferior-posterior positioning of hyoid. A small and shallow mandible is an independent risk factor for OSA in males but not in females.
Assuntos
Anormalidades Craniofaciais/complicações , Apneia Obstrutiva do Sono/etiologia , Adulto , Estudos de Casos e Controles , Cefalometria/métodos , Anormalidades Craniofaciais/fisiopatologia , Feminino , Humanos , Osso Hioide/anormalidades , Osso Hioide/fisiopatologia , Imageamento por Ressonância Magnética , Masculino , Mandíbula/anormalidades , Mandíbula/fisiopatologia , Pessoa de Meia-Idade , Tamanho do Órgão , Faringe/anormalidades , Faringe/fisiopatologia , Fatores de Risco , Fatores Sexuais , Apneia Obstrutiva do Sono/fisiopatologia , Língua/anormalidades , Língua/fisiopatologiaRESUMO
Dermal exposure to organophosphate pesticide is important because of its popular use. This study planned to compare the changes in serum acetylcholinesterase, paraoxonase and neuronal density of hippocampus and iso-cortex between two age groups of Swiss albino mice (18-day-old and 150-day-old) after dermal application of (1/2) LD50 of chlorpyrifos for 14 days. Statistically significant reduction was observed in serum acetylcholinesterase (Mann-Whitney test, p<0.05) and neuronal density (Independent samples t-test, p<0.05) in exposed groups compared to the control. The reduction in serum AChE and neuronal density was more pronounced in exposed adult mice than in exposed neonatal mice. The paraoxonase level was insignificant in control neonatal mice, whereas it was 890-fold more in exposed neonatal mice. Upregulated paraoxonase levels may be extrapolated to produce relatively lower reduction of cholinesterase and neuronal density in neonatal mice.
Assuntos
Doenças do Sistema Nervoso Central/induzido quimicamente , Clorpirifos/administração & dosagem , Clorpirifos/toxicidade , Inseticidas/administração & dosagem , Inseticidas/toxicidade , Acetilcolinesterase/sangue , Acetilcolinesterase/metabolismo , Administração Tópica , Animais , Animais Recém-Nascidos , Arildialquilfosfatase/sangue , Arildialquilfosfatase/metabolismo , Masculino , CamundongosRESUMO
The association between hypertension and chronic renal disease is well known. The pathogenesis of hypertension in patients with chronic kidney disease (CKD) is complex and multifactorial, which may explain why it is resistant to treatment. The traditional paradigm is that hypertension in CKD is due either to an excess of intravascular volume (volume dependent) or to excessive activation of the renin-angiotensin system in relation to the state of sodium/volume balance (renin-dependent hypertension). This review focuses on the importance of less established mechanisms, such as increased activity of the sympathetic nervous system, increased endothelin production, decreased availability of endothelium-derived vasodilators and structural changes of the arteries, renal ischemia, and sleep apnea.
Assuntos
Anti-Hipertensivos/uso terapêutico , Hipertensão/tratamento farmacológico , Hipertensão/fisiopatologia , Nefropatias/complicações , Nefropatias/fisiopatologia , Artérias/patologia , Artérias/fisiopatologia , Doença Crônica , Ritmo Circadiano/fisiologia , Resistência a Medicamentos/fisiologia , Endotelinas/fisiologia , Endotélio Vascular/fisiopatologia , Humanos , Hipertensão/etiologia , Doença Iatrogênica , Estresse Oxidativo/fisiologia , Sistema Renina-Angiotensina/fisiologia , Síndromes da Apneia do Sono/complicações , Síndromes da Apneia do Sono/fisiopatologia , Sistema Nervoso Simpático/fisiopatologiaRESUMO
Methylglyoxal (MGO), a potent glycating agent, forms advanced glycation end products (AGEs) with proteins. Several diabetic complications including cataract are thought to be the result of accumulation of these protein-AGEs. alpha-Crystallin, molecular chaperone of the eye lens, plays an important role in maintaining the transparency of the lens by preventing the aggregation/inactivation of several proteins/enzymes in addition to its structural role. Binding of adenosine-5-triphosphate (ATP) to alpha-crystallin has been shown to enhance its chaperone-like function and protection against proteolytic degradation. In the earlier study, we have shown that modification of alpha-crystallin by MGO caused altered chaperone-like activity along with structural changes, cross-linking, coloration and subsequent insolubilization leading to scattering of light [Biochem. J. 379 (2004) 273]. In the present study, we have investigated ATP binding, stability and degradation of MGO-modified alpha-crystallin. Proteolytic digestion with trypsin and chymotrypsin showed that MGO-modified alpha-crystallin is more susceptible to degradation compared to native alpha-crystallin. Furthermore, ATP was able to protect native alpha-crystallin against proteolytic cleavage but not MGO-modified alpha-crystallin. Interestingly, binding studies indicate decreased ATP binding to MGO-modified alpha-crystallin and support the decreased protection by ATP against proteolysis. In addition, differential scanning calorimetric and denaturant-induced unfolding studies indicate that modification of alpha-crystallin by MGO leads to decreased stability. These results indicate that MGO-modification of alpha-crystallin causes partial unfolding and decreased stability leading to enhanced proteolysis. Cross-linking of these degraded products could result in aggregation and subsequent insolubilization as observed in senile and diabetic cataract lenses.
Assuntos
Cristalino/química , Aldeído Pirúvico/farmacologia , alfa-Cristalinas/química , Trifosfato de Adenosina/química , Animais , Bovinos , Quimotripsina/química , Chaperonas Moleculares/química , Dobramento de Proteína , Relação Estrutura-Atividade , Tripsina/química , alfa-Cristalinas/efeitos dos fármacosRESUMO
The three-dimensional structure of the recombinant form of Erythrina corallodendron lectin, complexed with lactose, has been elucidated by X-ray crystallography at 2.55 A resolution. Comparison of this non-glycosylated structure with that of the native glycosylated lectin reveals that the tertiary and quaternary structures are identical in the two forms, with local changes observed at one of the glycosylation sites (Asn17). These changes take place in such a way that hydrogen bonds with the neighboring protein molecules in rECorL compensate those made by the glycan with the protein in ECorL. Contrary to an earlier report, this study demonstrates that the glycan attached to the lectin does not influence the oligomeric state of the lectin. Identical interactions between the lectin and the non-covalently bound lactose in the two forms indicate, in line with earlier reports, that glycosylation does not affect the carbohydrate specificity of the lectin. The present study, the first of its kind involving a glycosylated protein with a well-defined glycan and the corresponding deglycosylated form, provides insights into the structural aspects of protein glycosylation.
Assuntos
Lectinas de Plantas/química , Lectinas de Plantas/metabolismo , Sítios de Ligação , Metabolismo dos Carboidratos , Cristalografia por Raios X , Glicosilação , Lactose/química , Lactose/metabolismo , Lectinas/química , Lectinas/metabolismo , Estrutura Quaternária de Proteína , Estrutura Terciária de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismoRESUMO
Effect of chronic intake of alcohol and its subsequent withdrawal was studied in albino mice on the layers of neurons of the iso-cortex. Neuronal density per mm2 of section in different layers of iso-cortex was counted and compared in 3 groups of animals (control, ethanol fed and withdrawal). Qualitative changes on nissl granules of neurons and myelinated fibres were also studied. Mice fed with 10% ethanol v/v ad libitum for 6 months showed loss of nissl granules and nucleolus and discontinuity of nuclear membrane. Quantitatively, significant reduction in neuronal density (P<0.001) was observed in layers II+III IV and V neurons of iso-cortex. Withdrawal of ethanol for 2 months showed continued reduction of counts of neuronal density in layers II+III and V only whereas reversal of count was found significantly (P<0.001) in layer IV of iso-cortex. Qualitatively, only few neurons showed prominent nissl granules after withdrawal of ethanol. More afferent synaptic connection in layer IV may be suggested as probable factor helping relative replenishment of neuronal count after withdrawal of alcohol.
Assuntos
Depressores do Sistema Nervoso Central/toxicidade , Córtex Cerebral/efeitos dos fármacos , Etanol/toxicidade , Neurônios/efeitos dos fármacos , Síndrome de Abstinência a Substâncias/patologia , Animais , Córtex Cerebral/patologia , Feminino , Masculino , Camundongos , Neurônios/patologiaRESUMO
OBJECTIVE: The purpose of this study was to measure and compare the effect of d-alpha-tocopheryl succinate (alpha-TS) in modifying radiation-induced chromosomal damage in human normal cells and cancer cells in culture. METHODS: Three human normal fibroblast cell lines (GM2149, AG1522 and HF19) and three human cancer cell lines, cervical cancer (HeLa) and ovarian carcinoma cells (OVGI and SKOV3) were treated with alpha-TS (37.6 microM) 20 hours before 100 cGy gamma-irradiation. After 30 minutes of irradiation, colcemid was added and cells were fixed. One hundred randomly selected metaphase cells were scored for the presence of chromatid gaps and breaks. To study the cellular accumulation of alpha-TS. cells were incubated in the presence of alpha-TS (18.8 and 37.6 microM) for 24 hours, and alpha-TS was extracted with hexane using a-tocopheryl acetate as an internal standard. The levels of alpha-TS were determined by HPLC. RESULTS: Results showed that alpha-TS induced chromosomal damage in both human cervical cancer cells and ovarian cancer cells, but not in human normal fibroblasts in culture. In addition, alpha-TS enhanced the level of radiation-induced chromosomal damage in cancer cells, but it protected normal cells against such damage. Both cancer cells and normal cells accumulated similar levels of alpha-TS, suggesting that increased sensitivity of cancer cells to alpha-TS is acquired during transformation. CONCLUSION: The use of alpha-TS during radiation therapy may improve the efficacy of radiation therapy by enhancing tumor response and decreasing some of the toxicities on normal cells.
Assuntos
Dano ao DNA/efeitos dos fármacos , Raios gama/efeitos adversos , Protetores contra Radiação/farmacologia , alfa-Tocoferol/análogos & derivados , alfa-Tocoferol/farmacologia , Linhagem Celular , Cromatografia Líquida de Alta Pressão , Dano ao DNA/efeitos da radiação , Feminino , Fibroblastos/efeitos dos fármacos , Fibroblastos/patologia , Células HeLa , Humanos , Neoplasias Ovarianas/tratamento farmacológico , Neoplasias Ovarianas/patologia , Neoplasias Ovarianas/radioterapia , Tocoferóis , Células Tumorais Cultivadas , Neoplasias do Colo do Útero/tratamento farmacológico , Neoplasias do Colo do Útero/patologia , Neoplasias do Colo do Útero/radioterapiaRESUMO
It has been known for many years that caffeine reduces or eliminates the G2-phase cell cycle delay normally seen in human HeLa cells or Chinese hamster ovary (CHO) cells after exposure to X or gamma rays. In light of our recent demonstration of a consistent difference between human normal and tumor cells in a G2-phase checkpoint response in the presence of microtubule-active drugs, we examined the effect of caffeine on the G2-phase delays after exposure to gamma rays for cells of three human normal cell lines (GM2149, GM4626, AG1522) and three human tumor cell lines (HeLa, MCF7, OVGI). The G2-phase delays after a dose of 1 Gy were similar for all six cell lines. In agreement with the above-mentioned reports for HeLa and CHO cells, we also observed that the G2-phase delays were eliminated by caffeine in the tumor cell lines. In sharp contrast, caffeine did not eliminate or even reduce the gamma-ray-induced G2-phase delays in any of the human normal cell lines. Since caffeine has several effects in cells, including the inhibition of cAMP and cGMP phosphodiesterases, as well as causing a release of Ca(++) from intracellular stores, we evaluated the effects of other drugs affecting these processes on radiation-induced G2-phase delays in the tumor cell lines. Drugs that inhibit cAMP or cGMP phosphodiesterases did not eliminate the radiation-induced G2-phase delay either separately or in combination. The ability of caffeine to eliminate radiation-induced G2-phase delay was, however, partially reduced by ryanodine and eliminated by thapsigargin, both of which can modulate intracellular calcium, but by different mechanisms. To determine if caffeine was acting through the release of calcium from intracellular stores, calcium was monitored in living cells using a fluorescent calcium indicator, furaII, before and after the addition of caffeine. No calcium release was seen after the addition of caffeine in either OVGI tumor cells or GM2149 normal cells, even though a large calcium release was measured in parallel experiments with ciliary neurons. Thus it is likely that caffeine is eliminating the radiation-induced G2-phase delay through a Ca(++)-independent mechanism, such as the inhibition of a cell cycle-regulating kinase.
Assuntos
Cafeína/farmacologia , Fase G2/efeitos dos fármacos , Raios gama/efeitos adversos , Células-Tronco Neoplásicas/efeitos dos fármacos , 3',5'-AMP Cíclico Fosfodiesterases/antagonistas & inibidores , 3',5'-GMP Cíclico Fosfodiesterases/antagonistas & inibidores , 4-(3-Butoxi-4-metoxibenzil)-2-imidazolidinona/farmacologia , Animais , Neoplasias da Mama/patologia , Bloqueadores dos Canais de Cálcio/farmacologia , Sinalização do Cálcio/efeitos dos fármacos , Carcinoma/patologia , Galinhas , Demecolcina/farmacologia , Resistência a Medicamentos , Feminino , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/efeitos da radiação , Fase G2/efeitos da radiação , Células HeLa/citologia , Células HeLa/efeitos dos fármacos , Células HeLa/efeitos da radiação , Humanos , Mitose/efeitos dos fármacos , Mitose/efeitos da radiação , Células-Tronco Neoplásicas/citologia , Células-Tronco Neoplásicas/efeitos da radiação , Neurônios/efeitos dos fármacos , Neoplasias Ovarianas/patologia , Inibidores de Fosfodiesterase/farmacologia , Quinazolinas/farmacologia , Tolerância a Radiação/efeitos dos fármacos , Rianodina/farmacologia , Canal de Liberação de Cálcio do Receptor de Rianodina/efeitos dos fármacos , Tapsigargina/farmacologia , Células Tumorais Cultivadas/citologia , Células Tumorais Cultivadas/efeitos dos fármacos , Células Tumorais Cultivadas/efeitos da radiaçãoRESUMO
Legume lectins family of proteins, despite having the same 'jelly roll' tertiary structural fold at monomeric level, exhibit considerable variation in their quaternary structure arising out of small changes in their sequence. Nevertheless, their folding behavior and stability correlates very well with their patterns of assembly into dimers and tetramers. A conservation of their fold during evolution, its wide distribution in many protein families together with the availability of structural information on them make them interesting as proteins to explore the effect of inter- versus intra-subunit interactions in the stability of multimeric proteins. Additionally, as 'natural mutants' of quaternary association, proteins of legume lectin family provide interesting paradigms for studies addressing the effect of subunit oligomerization on the stability, folding and function as well as the evolution of multimeric structures.
Assuntos
Fabaceae/química , Lectinas/química , Plantas Medicinais , Sequência de Aminoácidos , Varredura Diferencial de Calorimetria , Dimerização , Modelos Moleculares , Dados de Sequência Molecular , Lectinas de Plantas , Desnaturação Proteica , Dobramento de Proteína , Alinhamento de SequênciaRESUMO
BACKGROUND: The authors used propensity score adjustment to investigate the impact of intensive screening on stage of breast carcinoma at diagnosis in women who were at elevated risk for breast carcinoma. METHODS: The authors compared 58 women participating in a surveillance program at the Columbia Presbyterian Medical Center of New York Presbyterian Hospital who developed breast carcinoma with 3022 nonparticipating breast carcinoma patients. A propensity score was constructed for each woman by using important background covariates, and multivariable regression modeling was used to estimate the association of program membership with disease stage after adjusting for the propensity score. RESULTS: Before propensity score adjustment, nine baseline covariates significantly differed between the two groups (number of pregnancies, number of births, age at first delivery, race, how the tumor was discovered, history of prior breast disease, breast carcinoma in mother, breast carcinoma in maternal aunt, and breast carcinoma in sister), and there was a significant difference in stage at diagnosis. After adjustment, no significant differences remained. Program participants were more likely to have lower stage tumors at diagnosis than nonparticipants, but this association did not reach statistical significance (odds ratio, 1.52; 95% confidence interval, 0.94--2.46). CONCLUSIONS: Propensity score methods can remove bias in treatment comparisons in observational studies. An intensive surveillance program at a major cancer center may have had some effect on improving stage at diagnosis, but this effect was not statistically significant.
Assuntos
Neoplasias da Mama/diagnóstico , Técnicas e Procedimentos Diagnósticos , Feminino , Humanos , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Observação/métodos , Análise de RegressãoRESUMO
To determine the molecular basis for the insensitivity of rat alpha(IIb)beta(3) to inhibition by RGD-containing peptides, hybrids of human and rat alpha(IIb)beta(3) and chimeras of alpha(IIb)beta(3) in which alpha(IIb) was composed of portions of human and rat alpha(IIb) were expressed in Chinese hamster ovary cells and B lymphocytes, and the ability of the tetrapeptide RGDS to inhibit fibrinogen binding to the various forms of alpha(IIb)beta(3) was measured. These measurements indicated that sequences regulating the sensitivity of alpha(IIb)beta(3) to RGDS are located in the seven amino-terminal repeats of alpha(IIb). Moreover, replacing the first three or four (but not the first two) repeats of rat alpha(IIb) with the corresponding human sequences enhanced sensitivity to RGDS, whereas replacing the first two or three repeats of human alpha(IIb) with the corresponding rat sequences had little or no effect. Nevertheless, RGDS bound to Chinese hamster ovary cells expressing alpha(IIb)beta(3) regardless whether the alpha(IIb) in the heterodimers was human, rat, or a rat-human chimera. These results indicate that the sequences determining the sensitivity of alpha(IIb)beta(3) to RGD-containing peptides are located in the third and fourth amino-terminal repeats of alpha(IIb). Because RGDS binds to both human and rat alpha(IIb)beta(3), the results suggest that differences in RGDS sensitivity result from differences in the allosteric changes induced in these repeats following RGDS binding.
Assuntos
Plaquetas/metabolismo , Fibrinogênio/química , Fibrinogênio/metabolismo , Oligopeptídeos/química , Complexo Glicoproteico GPIIb-IIIa de Plaquetas/química , Complexo Glicoproteico GPIIb-IIIa de Plaquetas/metabolismo , Difosfato de Adenosina/metabolismo , Sítio Alostérico , Animais , Linfócitos B/metabolismo , Células CHO , Carcinógenos , Adesão Celular , Cricetinae , DNA Complementar/metabolismo , Ditiotreitol/farmacologia , Relação Dose-Resposta a Droga , Humanos , Concentração Inibidora 50 , Ligantes , Agregação Plaquetária , Ligação Proteica , Estrutura Terciária de Proteína , Ratos , Proteínas Recombinantes de Fusão/química , Acetato de Tetradecanoilforbol , TransfecçãoRESUMO
RNase S consists of two proteolytic fragments of RNase A, residues 1-20 (S20) and residues 21-124 (S pro). A 15-mer peptide (S15p) with high affinity for S pro was selected from a phage display library. Peptide residues that are buried in the structure of the wild type complex are conserved in S15p though there are several changes at other positions. Isothermal titration calorimetry studies show that the affinity of S15p is comparable to that of the wild type peptide at 25 degrees C. However, the magnitudes of DeltaH(o) and DeltaC(p) are lower for S15p, suggesting that the thermal stability of the complex is enhanced. In agreement with this prediction, at pH 6, the T(m) of the S15p complex was found to be 10 degrees C higher than that of the wild type complex. This suggests that for proteins where fragment complementation systems exist, phage display can be used to find mutations that increase protein thermal stability.
Assuntos
Ribonucleases/química , Sequência de Aminoácidos , Animais , Sequência de Bases , Calorimetria , Sequência Conservada , Primers do DNA/genética , Estabilidade Enzimática , Humanos , Técnicas In Vitro , Dados de Sequência Molecular , Mutação , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/genética , Fragmentos de Peptídeos/metabolismo , Biblioteca de Peptídeos , Desnaturação Proteica , Ribonuclease Pancreático/química , Ribonuclease Pancreático/genética , Ribonuclease Pancreático/metabolismo , Ribonucleases/genética , Ribonucleases/metabolismo , Homologia de Sequência de Aminoácidos , Especificidade por Substrato , TermodinâmicaRESUMO
All opportunities for immunisation in children should be utilised properly, as sustaining high levels of full immunisation coverage is essential to meet the goal of eradication of poliomyelitis as well as control the other vaccine-preventable diseases; yet many opportunities for immunisation are missed in all types of health facilities, even in teaching hospitals. Reducing such missed opportunities is the cheapest way to increase immunisation coverage. The present study discusses the problem of missed opportunities for immunisation in children in Paediatric Outpatient Department and Immunisation Clinic of R.G. Kar Medical and Hospital, Calcutta and the underlying factors of the problem. Prevalence of missed opportunities in Paediatric OPD and Immunisation clinic was 37.8% and 1.4% respectively. Most of the missed opportunities were attributed to health care providers and delivery system of health care of the studied hospital.
Assuntos
Controle de Doenças Transmissíveis/normas , Fidelidade a Diretrizes/estatística & dados numéricos , Imunização/estatística & dados numéricos , Pré-Escolar , Feminino , Humanos , Índia/epidemiologia , MasculinoRESUMO
Adequate maternal vitamin A nutrition is essential for successful pregnancy outcome and estimation of serum retinol among pregnant women enables a precise and objective assessment of vitamin A status, during pregnancy, even in subclinical state. In order to evaluate vitamin A status during pregnancy, and its relationship with personal and pregnancy related variables of the mother, 300 antenatal clinic attenders were interviewed at RG Kar Medical College and Hospital, Calcutta and tested for serum retinol, using Carr-Price test. Serum retinol values less than 30 microg/dl and 20 microg/dl, in this study, were considered as poor and severe vitamin A deficiency respectively. According to this 14.7% and 4% pregnant women were found to be suffering from poor and severe vitamin A deficiency respectively. Clinical signs of vitamin A deficiency (eg, nightblindness) were reported only among 60% cases of the deficient population. The problems of vitamin A deficiency were associated with low literacy and poor nutritional status of the mother, advanced gestational age of current pregnancy,increased number of pregnancies, shorter interval between births and poor dietary intake of vitamin A rich foods during pregnancy. The study thus raises the question of supplementation of vitamin A, during pregnancy, in Indian context where habitual diets are either inadequate or deficient in vitamin A.
Assuntos
Comportamento Alimentar , Cegueira Noturna/epidemiologia , Política Nutricional , Complicações na Gravidez/epidemiologia , Deficiência de Vitamina A/epidemiologia , Vitamina A/sangue , Adolescente , Adulto , Suplementos Nutricionais , Comportamento Alimentar/etnologia , Feminino , Humanos , Incidência , Índia/epidemiologia , Cegueira Noturna/prevenção & controle , Gravidez , Complicações na Gravidez/prevenção & controle , Prevalência , Fatores de Risco , Estudos de Amostragem , Fatores Socioeconômicos , Deficiência de Vitamina A/sangue , Deficiência de Vitamina A/prevenção & controleRESUMO
A child of first-cousin Puerto Rican parents had global developmental delay, failure to thrive, and hypotonia since early infancy. At 1 1/2 years of age, she developed clinical and electrophysiologic evidence of progressive motor and sensory neuropathy. At 2 1/2 years, she developed visual impairment and optic atrophy followed by gradual involvement of the 7th, 9th, 10th, and 12th cranial nerves. Uncontrollable myoclonic seizures began at 4 years and she died at 6 years of age. Motor nerve conduction velocities were initially normal and later became markedly slowed. Sensory distal latency responses were absent. Lysosomal enzyme activities in leukocytes and fibroblasts were normal. Sural nerve and two muscle biopsies showed only nondiagnostic abnormalities. Electron microscopy of lymphocytes, skin, and fibroblasts showed cytoplasmic inclusions. Light microscopy of frontal cortex biopsy showed neuronal storage material staining positively with Luxol fast blue, and electron microscopy showed cytoplasmic membranous bodies in neurons, suggesting an accumulation of a ganglioside. At autopsy, all organs were small but otherwise normal and without abnormal storage cells in the liver, spleen, or bone marrow. Anterior spinal nerve roots showed loss of large myelinated axons. The brain was small and atrophic; cortical neurons showed widespread accumulation of storage material, most marked in the pyramidal cell layer of the hippocampus. Subcortical white matter was gliotic with loss of axons and myelin sheaths. In cortical gray matter there was a 35% elevation of total gangliosides, with a 16-fold increase in GM3, a three- to four-fold increase in GM2 gangliosides, and a 15-fold elevation of lactosyl ceramide. GM3 sialidase activity was normal in gray matter at 3.1 nmols/mg protein per hour and lactosyl ceraminidase I and II activities were 70% to 80% of normal. In white matter, total myelin was reduced by 50% but its composition was normal. Phospholipid distribution and sphingomyelin content were normal in gray matter, white matter, and in the liver. These biochemical findings were interpreted as nonspecific abnormalities. The nature of the neuronal storage substance remains to be determined.