ABSTRACT
BACKGROUND: The literature on antiplatelet therapy for peripheral artery disease has historically been summarized inconsistently, leading to conflict between international guidelines. An umbrella review and meta-analysis was performed to summarize the literature, allow assessment of competing safety risks and clinical benefits, and identify weak areas for future research. METHODS: MEDLINE, Embase, DARE, PROSPERO and Cochrane databases were searched from inception until January 2019. All meta-analyses of antiplatelet therapy in peripheral artery disease were included. Quality was assessed using AMSTAR scores, and GRADE analysis was used to quantify the strength of evidence. Data were pooled using random-effects models. RESULTS: Twenty-eight meta-analyses were included. Thirty-three clinical outcomes and 41 antiplatelet comparisons in 72 181 patients were analysed. High-quality evidence showed that antiplatelet monotherapy reduced non-fatal strokes (3 (95 per cent c.i. 0 to 6) fewer per 1000 patients), In symptomatic patients, it reduced cardiovascular deaths (8 (0 to 16) fewer per 1000 patients), but increased the risk of major bleeding (7 (3 to 14) more events per 1000). In asymptomatic patients, monotherapy reduced non-fatal strokes (5 (0 to 8) fewer per 1000), but had no other clinical benefit. Dual antiplatelet therapy caused more major bleeding after intervention than monotherapy (37 (8 to 102) more events per 1000), with very low-quality evidence of improved endovascular patency (risk ratio 4·00, 95 per cent c.i. 0·91 to 17·68). CONCLUSION: Antiplatelet monotherapy has minimal clinical benefit for asymptomatic peripheral artery disease, and limited benefit for symptomatic disease, with a clear risk of major bleeding. There is a lack of evidence to guide antiplatelet prescribing after peripheral endovascular intervention.
ANTECEDENTES: Históricamente, la literatura del tratamiento antiplaquetario en la enfermedad arterial periférica se ha sintetizado inconsistentemente, lo que ha dado lugar a divergencias entre las guías internacionales. Se efectuó una amplia revision con metaanálisis para sintetizar claramente la literatura, permitiendo evaluar los riesgos competitivos de seguridad y los beneficios clínicos, e identificar áreas poco claras susceptibles de futuras investigaciones. MÉTODOS: La búsqueda se realizó en las bases de datos MEDLINE, EMBASE, DARE, PROSPERO y Cochrane desde su inicio hasta enero de 2019. Se incluyeron todos los metaanálisis del tratamiento antiplaquetario en la enfermedad arterial periférica. Se estimó su calidad utilizando la puntuación Amstar y la consistencia de su evidencia mediante el sistema GRADE. Los datos se agruparon utilizando modelos de efectos aleatorios. RESULTADOS: Se incluyeron 28 metaanálisis. Se analizaron 33 resultados clínicos y 41 comparaciones antiplaquetarias en 72.181 pacientes. Una evidencia de alta calidad demostró que la monoterapia antiplaquetaria reducía los accidentes cerebrovasculares no mortales y la muerte cardiovascular en pacientes sintomáticos (3 y 8 veces menos por 1.000 pacientes, respectivamente, i.c. del 95% 0-6 y 0-16), pero aumentó el riesgo de hemorragia grave (7 veces más por 1.000, i.c. del 95% 3-14). En pacientes asintomáticos, la monoterapia redujo los accidentes cerebrovasculares no mortales (5 veces menos por 1.000, i.c. del 95% 0-8) sin otro beneficio clínico. El doble tratamiento antiplaquetario causó más hemorragias graves después de cualquier intervención que la monoterapia (37 veces más por 1.000, i.c. del 95% 8-102), con una evidencia de muy baja calidad acerca de la mejoría de la permeabilidad endovascular (riesgo relativo 4,00, i.c. del 95% 0,91-17,68). CONCLUSIÓN: La monoterapia antiplaquetaria tiene un beneficio clínico mínimo en la enfermedad arterial periférica asintomática y un beneficio limitado en la sintomática, con un claro riesgo de hemorragia grave. No existe evidencia para recomendar la prescripción de antiagregantes plaquetarios después de una intervención endovascular periférica, situación que debería abordarse en ensayos aleatorizados con una potencia estadística adecuada.
Subject(s)
Peripheral Arterial Disease/drug therapy , Platelet Aggregation Inhibitors/therapeutic use , Anticoagulants/therapeutic use , Cardiovascular Diseases/prevention & control , Drug Therapy, Combination , Hemorrhage/chemically induced , Humans , Randomized Controlled Trials as Topic , Secondary Prevention , Stroke/prevention & control , Treatment OutcomeABSTRACT
In 2013, the national surveillance case definition for West Nile virus (WNV) disease was revised to remove fever as a criterion for neuroinvasive disease and require at most subjective fever for non-neuroinvasive disease. The aims of this project were to determine how often afebrile WNV disease occurs and assess differences among patients with and without fever. We included cases with laboratory evidence of WNV disease reported from four states in 2014. We compared demographics, clinical symptoms and laboratory evidence for patients with and without fever and stratified the analysis by neuroinvasive and non-neuroinvasive presentations. Among 956 included patients, 39 (4%) had no fever; this proportion was similar among patients with and without neuroinvasive disease symptoms. For neuroinvasive and non-neuroinvasive patients, there were no differences in age, sex, or laboratory evidence between febrile and afebrile patients, but hospitalisations were more common among patients with fever (P < 0.01). The only significant difference in symptoms was for ataxia, which was more common in neuroinvasive patients without fever (P = 0.04). Only 5% of non-neuroinvasive patients did not meet the WNV case definition due to lack of fever. The evidence presented here supports the changes made to the national case definition in 2013.
Subject(s)
Asymptomatic Diseases/epidemiology , Fever/epidemiology , West Nile Fever/diagnosis , West Nile Fever/epidemiology , West Nile virus/isolation & purification , California/epidemiology , Clinical Laboratory Techniques/methods , Female , Fever/diagnosis , Humans , Incidence , Louisiana/epidemiology , Male , Massachusetts/epidemiology , Minnesota/epidemiology , Population Surveillance , Retrospective Studies , Risk Assessment , Severity of Illness IndexABSTRACT
BACKGROUND: The fermentation of dietary fiber to various organic acids is a beneficial function provided by the microbiota in the human large intestine. In particular, butyric acid contributes to host health by facilitating maintenance of epithelial integrity, regulating inflammation, and influencing gene expression in colonocytes. We sought to increase the concentration of butyrate in 20 healthy young adults through dietary supplementation with resistant starch (unmodified potato starch-resistant starch (RS) type 2). METHODS: Fecal samples were collected from individuals to characterize butyrate concentration via liquid chromatography and composition of the microbiota via surveys of 16S rRNA-encoding gene sequences from the Illumina MiSeq platform. Random Forest and LEfSe analyses were used to associate responses in butyrate production to features of the microbiota. RESULTS: RS supplementation increased fecal butyrate concentrations in this cohort from 8 to 12 mmol/kg wet feces, but responses varied widely between individuals. Individuals could be categorized into three groups based upon butyrate concentrations before and during RS: enhanced, high, and low (n = 11, 3, and 6, respectively). Fecal butyrate increased by 67 % in the enhanced group (from 9 to 15 mmol/kg), while it remained ≥11 mmol/kg in the high group and ≤8 mmol/kg in the low group. Microbiota analyses revealed that the relative abundance of RS-degrading organisms-Bifidobacterium adolescentis or Ruminococcus bromii-increased from ~2 to 9 % in the enhanced and high groups, but remained at ~1.5 % in the low group. The lack of increase in RS-degrading bacteria in the low group may explain why there was no increase in fecal butyrate in response to RS. The microbiota of individuals in the high group were characterized by an elevated abundance of the butyrogenic microbe Eubacterium rectale (~6 % in high vs. 3 % in enhanced and low groups) throughout the study. CONCLUSIONS: We document the heterogeneous responses in butyrate concentrations upon RS supplementation and identify characteristic of the microbiota that appear to underlie this variation. This study complements and extends other studies that call for personalized approaches to manage beneficial functions provided by gut microbiomes.
Subject(s)
Bacteria/classification , Butyric Acid/analysis , Intestine, Large/microbiology , Microbiota/drug effects , Starch/administration & dosage , Bacteria/drug effects , Dietary Supplements , Feces/microbiology , Female , High-Throughput Nucleotide Sequencing , Humans , Intestine, Large/metabolism , Male , RNA, Ribosomal, 16S/analysis , Starch/pharmacology , Young AdultABSTRACT
INTRODUCTION: The addictive aspect of smoking is well acknowledged. Research has shown that interventions by healthcare professionals have been shown to be effective and that smokers will benefit from smoking cessation counselling before, during and after their quit attempts. Dental hygienists, as part of the healthcare team, are well positioned to provide this counselling. MATERIAL AND METHODS: A questionnaire was completed by patients, staff, students and members of the public, during Mouth Cancer Awareness Day 2012 in the Dublin Dental University Hospital to assess the prevalence of smoking as well as the history of smoking and quit attempts by current and former smokers. RESULTS: The prevalence of smoking was lower than the national average. A total of 18.3% of those surveyed were smokers, 25% were former smokers, and 68% of the smokers had their first cigarette within 30 minutes of waking, indicating high dependence. DISCUSSION AND CONCLUSIONS: The majority of the smokers (79%) had attempted to quit. Stress was the most common reason for lapsing. The most common reasons for smoking cessation were health issues. The public is well disposed to receive information regarding smoking and the methods available to quit by healthcare professionals on health awareness days such as Mouth Cancer Awareness Day.
Subject(s)
Smoking Cessation/statistics & numerical data , Smoking/epidemiology , Adolescent , Adult , Aged , Health Promotion , Humans , Ireland/epidemiology , Middle Aged , Mouth Neoplasms/prevention & control , Prevalence , Tobacco Use Disorder/epidemiology , Young AdultABSTRACT
We have assessed the efficacy of anti retroviral therapy (ART) using undetectable viral load (VL) (<50 RNA copies/ml) as a marker of virological success, in patients who have Human Immunodeficiency Virus (HIV) attending the Department of Infectious Disease. A cross-sectional review of patients' case notes was used to obtain their demographics and treatment details. 79% (253) of the hospital case notes of clinic population was available for analysis, which represents 90% of those receiving ART in the clinic. 166/253 of the cohort were receiving treatment at the time of this study and 95% (157/166) of these were on treatment for greater than 6 months. The total virological success rate is 93%, which is comparable to other centres and are as good as those from published clinical trials. 56% of those on therapy who have virological failure were Intravenous Drug Users (IVDUs). Case by case investigation for those with treatment failure is warranted.
Subject(s)
Anti-HIV Agents/therapeutic use , HIV Infections/drug therapy , HIV-1/drug effects , Medical Audit , Adolescent , Adult , Anti-HIV Agents/pharmacology , CD4 Lymphocyte Count , Cross-Sectional Studies , Female , HIV Infections/transmission , HIV Infections/virology , Humans , Infectious Disease Medicine , Ireland , Male , Middle Aged , Retrospective Studies , Viral Load/drug effects , Young AdultABSTRACT
Rhamnose is an essential component of the insect control agent spinosad. However, the genes coding for the four enzymes involved in rhamnose biosynthesis in Saccharopolyspora spinosa are located in three different regions of the genome, all unlinked to the cluster of other genes that are required for spinosyn biosynthesis. Disruption of any of the rhamnose genes resulted in mutants with highly fragmented mycelia that could survive only in media supplemented with an osmotic stabilizer. It appears that this single set of genes provides rhamnose for cell wall synthesis as well as for secondary metabolite production. Duplicating the first two genes of the pathway caused a significant improvement in the yield of spinosyn fermentation products.
Subject(s)
Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Macrolides , Rhamnose/biosynthesis , Saccharopolyspora/enzymology , Alcohol Oxidoreductases/genetics , Alcohol Oxidoreductases/metabolism , Amino Acid Sequence , Anti-Bacterial Agents/biosynthesis , Cloning, Molecular , Gene Deletion , Gene Duplication , Glucosyltransferases/genetics , Glucosyltransferases/metabolism , Hydro-Lyases/genetics , Hydro-Lyases/metabolism , Molecular Sequence Data , Racemases and Epimerases/genetics , Racemases and Epimerases/metabolism , Rhamnose/genetics , Saccharopolyspora/genetics , Sequence Analysis, DNAABSTRACT
BACKGROUND: Spinosad is a mixture of novel macrolide secondary metabolites produced by Saccharopolyspora spinosa. It is used in agriculture as a potent insect control agent with exceptional safety to non-target organisms. The cloning of the spinosyn biosynthetic gene cluster provides the starting materials for the molecular genetic manipulation of spinosad yields, and for the production of novel derivatives containing alterations in the polyketide core or in the attached sugars. RESULTS: We cloned the spinosad biosynthetic genes by molecular probing, complementation of blocked mutants, and cosmid walking, and sequenced an 80 kb region. We carried out gene disruptions of some of the genes and analyzed the mutants for product formation and for the bioconversion of intermediates in the spinosyn pathway. The spinosyn gene cluster contains five large open reading frames that encode a multifunctional, multi-subunit type I polyketide synthase (PKS). The PKS cluster is flanked on one side by genes involved in the biosynthesis of the amino sugar forosamine, in O-methylations of rhamnose, in sugar attachment to the polyketide, and in polyketide cross-bridging. Genes involved in the early common steps in the biosynthesis of forosamine and rhamnose, and genes dedicated to rhamnose biosynthesis, were not located in the 80 kb cluster. CONCLUSIONS: Most of the S. spinosa genes involved in spinosyn biosynthesis are found in one 74 kb cluster, though it does not contain all of the genes required for the essential deoxysugars. Characterization of the clustered genes suggests that the spinosyns are synthesized largely by mechanisms similar to those used to assemble complex macrolides in other actinomycetes. However, there are several unusual genes in the spinosyn cluster that could encode enzymes that generate the most striking structural feature of these compounds, a tetracyclic polyketide aglycone nucleus.
Subject(s)
Cloning, Molecular , Macrolides/metabolism , Multienzyme Complexes/genetics , Multigene Family/genetics , Mutagenesis, Insertional/genetics , Saccharopolyspora/genetics , Amino Acid Sequence/genetics , Drug Combinations , Hexosamines/biosynthesis , Molecular Sequence Data , Multienzyme Complexes/metabolism , Open Reading Frames/genetics , Rhamnose/biosynthesis , Rhamnose/chemistry , Saccharopolyspora/chemistry , Saccharopolyspora/metabolismABSTRACT
Spinosyns A and D are the active ingredients in an insect control agent produced by fermentation of Saccharopolyspora spinosa. Spinosyns are macrolides with a 21-carbon, tetracyclic lactone backbone to which the deoxysugars forosamine and tri-O-methylrhamnose are attached. The spinosyn biosynthesis genes, except for the rhamnose genes, are located in a cluster that spans 74 kb of the S. spinosa genome. DNA sequence analysis, targeted gene disruptions and bioconversion studies identified five large genes encoding type I polyketide synthase subunits, and 14 genes involved in sugar biosynthesis, sugar attachment to the polyketide or cross-bridging of the polyketide. Four rhamnose biosynthetic genes, two of which are also necessary for forosamine biosynthesis, are located outside the spinosyn gene cluster. Duplication of the spinosyn genes linked to the polyketide synthase genes stimulated the final step in the biosynthesis--the conversion of the forosamine-less pseudoaglycones to endproducts. Duplication of genes involved in the early steps of deoxysugar biosynthesis increased spinosyn yield significantly.
Subject(s)
Anti-Bacterial Agents/biosynthesis , Bacterial Proteins/genetics , Genetic Engineering/methods , Multigene Family , Saccharopolyspora/genetics , Saccharopolyspora/metabolism , Bacterial Proteins/metabolism , Hexosamines/metabolism , Macrolides , Rhamnose/analogs & derivatives , Rhamnose/metabolismABSTRACT
Spinosyns A and D are the active ingredients in a family of insect control agents produced by fermentation of Saccharopolyspora spinosa. Spinosyns are 21-carbon tetracyclic lactones to which are attached two deoxysugars. Most of the genes involved in spinosyn biosynthesis are clustered in an 74 kb region of the S. spinosa genome. This region has been characterized by DNA sequence analysis and by targeted gene disruptions. The spinosyn biosynthetic gene cluster contains five large genes encoding a type I polyketide synthase, and 14 genes involved in modification of the macrolactone, or in the synthesis, modification and attachment of the deoxysugars. Four genes required for rhamnose biosynthesis (two of which are also required for forosamine biosynthesis) are not present in the cluster. A pathway for the biosynthesis of spinosyns is proposed.
Subject(s)
Anti-Bacterial Agents/biosynthesis , Multigene Family , Saccharopolyspora/genetics , Animals , Anti-Bacterial Agents/chemistry , Genes, Bacterial , Insecta , Lactones/chemistry , Macrolides , Molecular Conformation , Molecular Structure , Open Reading Frames , Pest Control, Biological , Saccharopolyspora/metabolism , Sequence Analysis, DNASubject(s)
Antipsychotic Agents/therapeutic use , Cardiovascular Agents/therapeutic use , Diuretics/therapeutic use , Drug Utilization/classification , Gastrointestinal Agents/therapeutic use , Medicare/statistics & numerical data , Drug Utilization/statistics & numerical data , Fee-for-Service Plans , Female , Health Benefit Plans, Employee , Health Maintenance Organizations , Health Services Research , Humans , Male , Medicaid , United StatesABSTRACT
There is increasing evidence that the distribution of monovalent cations in cardiac cells may be non-uniform, particularly in the region immediately beneath the sarcolemma, and we have proposed that a build-up of sodium in this region could be an important factor in the development of ischaemia-reperfusion injury. Electron probe X-ray microanalysis is ideal for the study of such changes in distribution but the application of the technique to this problem imposes severe requirements on the specimen and on the method for cryofixation. The specimen must be perfused through its vasculature so that it can be made truly ischaemic and be successfully reperfused. It is necessary to be able to cryofix the specimen without disturbance of its blood supply, electrical stimulation or temperature. It is also important to know the time in the contraction cycle when cryofixation occurs. Here we describe the design of an automated cryofixation device which can be used to cryofix a blood perfused papillary muscle preparation at predetermined time points in the contraction cycle. Preliminary data obtained from the analysis of rabbit papillary muscles subjected to varying periods of ischaemia are included as an example of the use of the cryoclamp.
Subject(s)
Cryopreservation/instrumentation , Cryopreservation/methods , Myocardial Reperfusion Injury/pathology , Papillary Muscles/ultrastructure , Animals , Cations/metabolism , Electron Probe Microanalysis , Equipment Design , In Vitro Techniques , Myocardial Contraction , Myocardial Reperfusion Injury/physiopathology , Papillary Muscles/physiology , RabbitsABSTRACT
The distinguishing histopathologic features of focal cemento-osseous dysplasia (FCOD) (including lesions occurring in both anterior and posterior jaws) and cemento-ossifying fibroma (COF) (ossifying fibroma and cementifying fibroma) were demonstrated in our earlier work. The aim of the current study was to further refine their clinical and radiographic features. We have assessed 18 clinical and radiographic parameters by univariate comparisons (chi-squared and Student t tests), and a multivariate assessment (logistic regression) in 241 cases of FCOD and 75 of COF. These cases were diagnosed from a combination of clinical, radiographic, and histopathologic information. FCOD was seen predominantly in black women, with a peak incidence in the fourth and fifth decades, whereas COF showed no female predilection except in the fourth decade (p < 0.005). COF occurred in patients an average of 10 years younger than patients with FCOD (p < 0.0001). Most patients with FCOD were asymptomatic (62%); the average lesion size was 1.8 cm. More than half of patients with COF displayed jaw expansion and a considerably larger size lesion (mean 3.8 cm, p < 0.001). The mandible was the most frequent site for both FCOD (86%) and COF (70%). Radiographically, a well-defined border was observed in 53% of cases of FCOD and 85% of cases of COF (p < 0.01). Cases of FCOD mostly demonstrated an irregularly mixed radio-opacity (69%), whereas 53% of COFs presented as a radiolucency (p < 0.005). In FCOD, there was a close association with tooth apices (70.6%, p < 0.0001) or with previous extraction sites (21%, p < 0.05); however, the majority of COF cases (86%) showed no relationship with either. Combining the radiographic feature of a periapical location with the pathology of multiple curetted fragments and "ginger root" bony trabeculae, allowed 90% sensitivity and 89% specificity in a logistic regression model to predict the lesion to be an FCOD. These findings provide guidelines not only to distinguish these two entities clinically, but also aid in reaching an accurate diagnosis histopathologically.
Subject(s)
Bone Diseases, Developmental/pathology , Fibroma, Ossifying/pathology , Fibrous Dysplasia of Bone/pathology , Jaw Diseases/pathology , Jaw Neoplasms/pathology , Odontogenic Tumors/pathology , Adolescent , Adult , Age Factors , Aged , Bone Diseases, Developmental/diagnostic imaging , Chi-Square Distribution , Child , Dental Cementum/diagnostic imaging , Dental Cementum/pathology , Diagnosis, Differential , Female , Fibroma, Ossifying/diagnostic imaging , Fibrous Dysplasia of Bone/diagnostic imaging , Humans , Jaw Diseases/diagnostic imaging , Jaw Neoplasms/diagnostic imaging , Logistic Models , Male , Mandibular Diseases/diagnostic imaging , Mandibular Diseases/pathology , Mandibular Neoplasms/diagnostic imaging , Mandibular Neoplasms/pathology , Middle Aged , Multivariate Analysis , Odontogenic Tumors/diagnostic imaging , Periapical Diseases/diagnostic imaging , Periapical Diseases/pathology , Radiography , Sensitivity and Specificity , Sex Factors , Tooth Apex/diagnostic imaging , Tooth Apex/pathology , Tooth ExtractionABSTRACT
Focally expressed cemento-osseous dysplasia (periapical cemento-osseous dysplasia and focal cemento-osseous dysplasia) and cemento-ossifying fibroma (ossifying fibroma and cementifying fibroma) are two clinically recognized entities that are not easily differentiated histopathologically because of the lack of recognition of specific microscopic features. We have assessed 20 pathologic parameters for their ability to distinguish reliably between the two. Included in this study were 241 cases of focally expressed cemento-osseous dysplasia and 75 cases of cemento-osseous fibroma diagnosed from a combination of clinical, radiographic, and histopathologic information. Results revealed that 92.5% of focally expressed cemento-osseous dysplasia were composed of multiple small fragments of tissue whereas 88.0% of cemento-osseous fibromas showed a large intact specimen. Thick curvilinear trabeculae ("ginger root" pattern) or irregularly shaped cementum-like masses were typically seen in focally expressed cemento-osseous dysplasia, whereas thin isolated trabeculae with prominent osteoblastic rimming were more commonly observed in cemento-osseous fibroma. The stroma of focally expressed cemento-osseous dysplasia often displayed characteristic cavernous-like vascularity that was almost always associated with bony trabeculae. Free hemorrhage was frequently interspersed in the artifactual spaces throughout focally expressed cemento-osseous dysplasia. In contrast, the cases of cemento-osseous fibroma showed more cellularity in the stroma in which a storiform pattern was present in more than half the lesions studied. Giant cells, when present in cemento-osseous fibroma, were clustered in the center of the cellular stroma. The features described here allowed distinction histopathologically in 94% of cases studied. Three progressive stages of focally expressed cemento-osseous dysplasia and subtypes of cemento-osseous fibroma may be recognizable microscopically.
Subject(s)
Bone Diseases, Developmental/pathology , Fibroma, Ossifying/pathology , Fibrous Dysplasia of Bone/pathology , Jaw Diseases/pathology , Jaw Neoplasms/pathology , Odontogenic Tumors/pathology , Blood Vessels/pathology , Bone Diseases, Developmental/diagnostic imaging , Bone and Bones/pathology , Cementoma/pathology , Dental Cementum/pathology , Diagnosis, Differential , Fibroma, Ossifying/diagnostic imaging , Fibrous Dysplasia of Bone/diagnostic imaging , Giant Cells/pathology , Hemorrhage/pathology , Humans , Jaw Diseases/diagnostic imaging , Jaw Neoplasms/diagnostic imaging , Logistic Models , Odontogenic Tumors/diagnostic imaging , Osteoblasts/pathology , Periapical Diseases/diagnostic imaging , Periapical Diseases/pathology , Radiography, Panoramic , Reproducibility of ResultsABSTRACT
The target site of the antifungal compound LY214352 [8-chloro-4-(2-chloro-4-fluorophenoxy) quinoline] has been identified through a dual biochemical and molecular-genetics approach. In the molecular-genetics approach, a cosmid library was prepared from an Aspergillus nidulans mutant that was resistant to LY214352 because of a dominant mutation in a single gene. A single cosmid (6A6-6) that could transform an LY214352-sensitive strain of A. nidulans to LY214352-resistance was isolated from the library by sib-selection. Restriction fragments from cosmid 6A6-6 containing the functional resistance gene were identified by transformation, and sequenced. The LY214352-resistance gene coded for a protein of 520 amino acids that had a 34% identity and a 57% similarity in a 333 amino-acid overlap to E. coli dihydroorotate dehydrogenase (DHO-DH). The results of a series of biochemical mechanism-of-action studies initiated simultaneously with molecular-genetic experiments also suggested that DHO-DH was the target of LY214352. Assays measuring the inhibition of DHO-DH activity by LY214352 in a wild-type strain (I50=40 ng/ml) and a highly resistant mutant (I50>100 microgram/ml) conclusively demonstrated that DHO-DH is the target site of LY214352 in A. nidulans. Several mutations in the DHO-DH (pyrE) gene that resulted in resistance to LY214352 were identified.
Subject(s)
Antifungal Agents/pharmacology , Aspergillus nidulans/genetics , Oxidoreductases Acting on CH-CH Group Donors , Oxidoreductases/genetics , Quinolines/pharmacology , Antifungal Agents/chemistry , Aspergillus nidulans/drug effects , Aspergillus nidulans/enzymology , Base Sequence , Cloning, Molecular , Cosmids , DNA, Fungal , Dihydroorotate Dehydrogenase , Drug Resistance, Microbial/genetics , Molecular Sequence Data , Protoplasts , Quinolines/chemistryABSTRACT
With an increase in breast-sparing surgery and adjuvant radiotherapy, there has been a concomitant increase in the complexity of breast reconstruction. The effects of radiotherapy on the internal mammary artery were evaluated with respect to flap viability for conventional transverse rectus abdominis musculocutaneous breast reconstruction using the irradiated rectus muscle. Twenty-eight women who received postoperative irradiation for breast cancer were studied. All women had unilateral irradiation, and evaluation of the internal mammary arteries was performed at least 1 year after the completion of radiotherapy to allow for fibrosis and long-term vascular changes. Examination of the internal mammary artery (upper and lower chains) was performed using color Doppler sonography to assess vessel diameter, peak systolic velocity, and blood flow. The nonirradiated side was compared to the radiated internal mammary artery as an internal control. Based on the details of prior radiotherapy, the 28 women were designated into two groups. Group I included 14 women with radiation portals that specifically treated the internal mammary chain; the average dose to the internal mammary chain was 47.44 Gy. Group II comprised 14 patients that received tangential portals that did not directly target the internal mammary chain region; the average radiation dose was 48.21 Gy to the chest wall. No statistical difference was observed in group I between the irradiated and nonirradiated side for vessel diameter (p = .8631) or peak systolic velocity (p = .2646). However, an increase in blood flow on the irradiated side was significant (p = .0321).(ABSTRACT TRUNCATED AT 250 WORDS)
Subject(s)
Breast Neoplasms/radiotherapy , Mammaplasty , Mammary Arteries/radiation effects , Mastectomy , Surgical Flaps , Adult , Breast Neoplasms/surgery , Female , Humans , Mammary Arteries/diagnostic imaging , Radiotherapy/adverse effects , Ultrasonography, Doppler, ColorABSTRACT
We have examined the relative roles of the calcium-calmodulin system and protein kinase C in angiotensin-mediated aldosterone secretion. We used a highly specific protein kinase C inhibitor, calphostin C and two well-accepted calmodulin inhibitors, W-7 and calmidazolium. Although both types of inhibitors affected angiotensin-induced aldosterone secretion, as judged by the inhibitory doses of these compounds, angiotensin-evoked aldosterone secretion was more sensitive to calmodulin inhibition than protein kinase C inhibition. Manipulation of OFFracellular calcium by dantrolene and thapsigargin also modified aldosterone secretion significantly.
Subject(s)
Aldosterone/metabolism , Angiotensin II/pharmacology , Calmodulin/antagonists & inhibitors , Naphthalenes , Polycyclic Compounds/pharmacology , Protein Kinase C/antagonists & inhibitors , Animals , Calcium/physiology , Cattle , Secretory Rate/drug effects , Sulfonamides/pharmacology , Zona Glomerulosa/enzymologyABSTRACT
Giant cell lesions of the maxilla and paranasal sinuses represent a rare, locally aggressive disorder which present as a soft tissue mass with distinct histologic and clinical features. There is considerable controversy on the therapeutic modalities, the prediction of clinical behavior based on histologic features, and whether these growths are reactive or neoplastic in nature. We present our clinical experience with four of these lesions. Follow-up ranged from 3 to 11 years. Our results and the rationale for aggressive surgical treatment will be discussed. We hypothesize that giant cell "granuloma" of the maxilla and paranasal sinuses and giant cell tumor of other bones represent a continuum of a single disease process, which may have an aggressive clinical behavior. This paper, with a literature review, will address the treatment controversy and advocate surgical resection for all giant cell lesions.
Subject(s)
Giant Cell Tumor of Bone/pathology , Granuloma, Giant Cell/pathology , Maxillary Diseases/pathology , Maxillary Neoplasms/pathology , Osteitis Deformans/pathology , Paranasal Sinus Diseases/pathology , Paranasal Sinus Neoplasms/pathology , Adult , Female , Follow-Up Studies , Giant Cells/pathology , Humans , Male , Middle Aged , Neoplasm Recurrence, Local , RecurrenceABSTRACT
A gene coding for potato multicystatin (PMC), the crystalline inhibitor of cysteine proteases which is found in tubers, was isolated and characterized. The deduced polypeptide product of this genomic sequence is 757 amino acids long and has a molecular mass of 86,778 Da. It consists exclusively of eight closely related domains, with 53-89% identity of residues. Each repeated unit is homologous to the cystatin superfamily of cysteine protease inhibitors. To date, no other member of this family has been found to contain so many inhibitor domains in one polypeptide. Eight introns are proposed in the 3.5 kb of genomic DNA coding for PMC, one in each cystatin unit. There is a family of 4 to 6 such large genes in potato, while in pea and maize the homologues are much smaller, and probably code for single-domain cystatins. PMC transcripts are abundant in tubers, but scarce in undamaged leaves or stems of field-grown potatoes. The tuber messages are derived from at least four genes (including the cloned example). The pattern of gene expression, as well as the properties of the protein, suggest that PMC has a role in the plant's defense system.
Subject(s)
Cystatins/genetics , Cysteine Proteinase Inhibitors/genetics , Genes, Plant , Plant Proteins/genetics , Solanum tuberosum/genetics , Amino Acid Sequence , Base Sequence , Cloning, Molecular , Consensus Sequence , DNA Primers/chemistry , DNA, Complementary/genetics , Molecular Sequence Data , Sequence Alignment , Sequence Homology, Amino AcidABSTRACT
PURPOSE: The authors clinically evaluated a new high-frequency ultrasound (US) scanner to determine the value of US for dermatologic applications. MATERIALS AND METHODS: A 20-MHz US scanner was used to visualize normal skin at eight sites in 10 healthy volunteers and to evaluate 200 skin lesions (45 malignant, 155 benign). RESULTS: In normal skin, the dermis was markedly echogenic and sharply demarcated from hypoechoic subcutaneous fat. The epidermis was not resolved except on the palm and sole. Only three superficial lesions were not identified with US; evaluation of another three was limited by shadowing. Thickness of the lesions visualized was 0.2-26.0 mm (mean, 1.9 mm +/- 2.6). Most lesions (77%) were hypoechoic, 9% were anechoic, 12% had mixed echogenicity, and 2% were isoechoic or hyperechoic. CONCLUSION: The diagnostic role of high-frequency US appears limited. It did not help differentiate benign from malignant lesions, but it did enable accurate delineation of deep margins of lesions and allowed noninvasive measurement of thickness. These features may help in the preoperative evaluation of skin tumors and in monitoring the response to therapy for certain inflammatory conditions.