Role of inflammation, oxidative stress, and autonomic nervous system activation during the development of right and left cardiac remodeling in experimental pulmonary arterial hypertension.

This study investigated the impact of experimental pulmonary arterial hypertension (PAH) progression by evaluating morphometric and functional parameters, oxidative stress, autonomic nervous system (ANS) activation, and inflammation in the right (RV) and left (LV) ventricles. Male rats were first divided into two groups: monocrotaline (MCT) and control. The MCT group received a single MCT injection (60 mg/kg, intraperitoneal), while control received saline. The MCT and control groups were further divided into four cohorts based on how long they were observed: 1, 2, 3, and 4 weeks. Animals were submitted to echocardiographic and hemodynamic analysis. RV and LV were used for morphometric, biochemical, and histological measurements. Autonomic modulation was evaluated by cardiac spectral analysis, considering two components: low frequency (LF) and high frequency (HF). Lung and liver weight was used for morphometric analysis. MCT induced 100% mortality at 4 weeks. In the RV, disease progression led to mild inflammation and enhanced reactive oxygen species (ROS) in week 1, followed by moderate inflammation, ROS production, and hypertrophy in week 2. By week 3, there was moderate inflammation, oxidative stress, and ANS imbalance, with development of right heart dysfunction. LV biochemical changes and inflammation were observed at week 3. The initial changes appeared to be related to inflammation and ROS, and the later ones to inflammation, oxidative stress, and ANS imbalance in MCT animals. This study reinforces the severity of the disease in the RV, the late effects in the LV, and the role of ANS imbalance in the development of heart dysfunction.

Electronic health records: new opportunities for clinical research.

Clinical research is on the threshold of a new era in which electronic health records (EHRs) are gaining an important novel supporting role. Whilst EHRs used for routine clinical care have some limitations at present, as discussed in this review, new improved systems and emerging research infrastructures are being developed to ensure that EHRs can be used for secondary purposes such as clinical research, including the design and execution of clinical trials for new medicines. EHR systems should be able to exchange information through the use of recently published international standards for their interoperability and clinically validated information structures (such as archetypes and international health terminologies), to ensure consistent and more complete recording and sharing of data for various patient groups. Such systems will counteract the obstacles of differing clinical languages and styles of documentation as well as the recognized incompleteness of routine records. Here, we discuss some of the legal and ethical concerns of clinical research data reuse and technical security measures that can enable such research while protecting privacy. In the emerging research landscape, cooperation infrastructures are being built where research projects can utilize the availability of patient data from federated EHR systems from many different sites, as well as in international multilingual settings. Amongst several initiatives described, the EHR4CR project offers a promising method for clinical research. One of the first achievements of this project was the development of a protocol feasibility prototype which is used for finding patients eligible for clinical trials from multiple sources.

[Mini-Mental State Examination in geriatrics : An evaluation of diagnostic quality]. / Mini-Mental-Status-Test im stationären geriatrischen Bereich.

BACKGROUND: Several studies have identified moderate reliability and validity for the Mini-Mental State Examination (MMSE). Some researchers showed the superiority of other dementia screening tests over the MMSE considering the test quality criteria. The aim of this study was the evaluation of MMSE, especially in the area of geriatrics. PATIENTS AND METHODS: MMSE and DemTect were carried out with 154 geriatric patients: 71 persons without cognitive impairment and 83 persons without delirium showed cognitive impairments as revealed by the DemTect. In addition, we also applied the Clock-Drawing-Test (CDT), Reisberg-Scale, Geriatric Depression-Scale (GDS, 15-item version) and the Confusion-Assessment-Method (CAM). RESULTS: According to the multitrait-multimethod approach, MMSE's convergent and divergent validity is similar to that of the DemTect. Both tests correlate only moderately with Spearman (r = 0.609) and revealed similar results for dementia in 57.1 % of the patients. MMSE showed low reliability and moderate reliability (Cronbach's α = 0.82) when ten items with low discriminatory power were excluded from the total test score. Difficulty of all items is only moderate (p = 0.86) and only eight items of the MMSE showed good test difficulty. CONCLUSION: All in all, DemTect and MMSE are not interchangeable. The MMSE estimates the average cognitive impairment of patients as considerably less pronounced than the DemTect. MMSE is, thus, not an instrument that would be recommended for the identification of mild cognitive impairment. In this case, tests with higher reliability and validity should be used.

Domestic animal models for biomedical research.

Experimental animals in biomedical research provide insights into disease mechanisms and models for determining the efficacy and safety of new therapies and for discovery of corresponding biomarkers. Although mouse and rat models are most widely used, observations in these species cannot always be faithfully extrapolated to human patients. Thus, a number of domestic species are additionally used in specific disease areas. This review summarizes the most important applications of domestic animal models and emphasizes the new possibilities genetic tailoring of disease models, specifically in pigs, provides.

Diagnosis and management of a fracture of the lateral trochlear ridge of the talus in a dog.

An eight-month-old, 31.2 kg, entire male Golden Retriever was presented for evaluation because it had a four-week history of right hindlimb lameness and audible popping occurring in association with movement of the right hindlimb. Mild right hindlimb lameness was noted upon gait analysis. Moderate to severe effusion and pain on extension were appreciated on palpation of the right tarsus. Dorsoplantar and lateral radiographs of the right tarsus revealed significant soft tissue swelling over the lateral aspect of the tarsus and widening of the joint space over the lateral trochlear ridge of the talus. A fracture of the lateral ridge was appreciated upon evaluation of the flexed dorsoplantar view. Un-enhanced computed tomography of the right tarsus confirmed fracture of the talus; one large and two small bone fragments were noted adjacent to the lateral aspect of the lateral trochlear ridge and medial to the fibula. The fracture was surgically repaired via a lateral approach; a fibular osteotomy was performed. The fragment was manually reduced and secured with a 1.5 mm cortical screw placed in lag fashion. The fibular osteotomy site was reduced and fixed with two 1.6 mm Kirshner wires and a tension band composed of 0.9 mm cerclage wire. The patient made a complete clinical recovery, however, the union was fibrous and evidence of mild osteo- arthritis was noted on postoperative radiographs.

Osteomyelitis-related sequestrum formation in association with the combination tibial plateau levelling osteotomy and cranial closing wedge osteotomy procedure.

A twenty-eight-month old female spayed American Bulldog was presented for evaluation of a chronic draining tract and intermittent left hindlimb lameness twenty-eight weeks after a combination tibial plateau levelling osteotomy and cranial closing wedge osteotomy (TLPO/CCWO) had been performed. The patient had developed an infection of the surgical site three weeks postoperatively. Drainage persisted despite implant removal 10 weeks postoperatively and several weeks of culture and sensitivity-directed antibiotic therapy. Twenty-eight weeks postoperatively, a sequestrum was identified on radiographs. Surgical removal of the sequestrum resulted in resolution of the drainage. While osteomyelitis is a known complication of TPLO surgery, this case represents the first described case of osteomyelitis-related sequestrum formation in association with the combined TPLO/CCWO procedure.

The product of the Saccharomyces cerevisiae RSS1 gene, identified as a high-copy suppressor of the rat7-1 temperature-sensitive allele of the RAT7/NUP159 nucleoporin, is required for efficient mRNA export.

RAT7/NUP159 was identified previously in a screen for genes whose products are important for nucleocytoplasmic export of poly(A)+ RNA and encodes an essential nucleoporin. We report here the identification of RSS1 (Rat Seven Suppressor) as a high-copy extragenic suppressor of the rat7-1 temperature-sensitive allele. Rss1p encodes a novel essential protein of 538 amino acids, which contains an extended predicted coiled-coil domain and is located both at nuclear pore complexes (NPCs) and in the cytoplasm. RSS1 is the first reported high-copy extragenic suppressor of a mutant nucleoporin. Overexpression of Rss1p partially suppresses the defects in nucleocytoplasmic export of poly(A)+ RNA, rRNA synthesis and processing, and nucleolar morphology seen in rat7-1 cells shifted to the nonpermissive temperature of 37 degrees C and, thus, restores these processes to levels adequate for growth at a rate approximately one-half that of wild-type cells. After a shift to 37 degrees C, the mutant Rat7-1p/Nup159-1p is lost from the nuclear rim of rat7-1 cells and NPCs, which are clustered together in these cells grown under permissive conditions become substantially less clustered. Overexpression of Rss1p did not result in retention of the mutant Rat7-1p/Nup159-1p in NPCs, but it did result in partial maintenance of the NPC-clustering phenotype seen in mutant cells. Depletion of Rss1p by placing the RSS1 open reading frame (ORF) under control of the GAL1 promoter led to cessation of growth and nuclear accumulation of poly(A)+ RNA without affecting nuclear protein import or nuclear pore complex distribution, suggesting that RSS1 is directly involved in mRNA export. Because both rat7-1 cells and cells depleted for Rss1p are defective in mRNA export, our data are consistent with both gene products playing essential roles in the process of mRNA export and suggest that Rss1p overexpression suppresses the growth defect of rat7-1 cells at 37 degrees C by acting to maintain mRNA export.

Numerical dosimetry ELF: accuracy of the method, variability of models and parameters, and the implication for quantifying guidelines.

In situ electric fields and current densities are investigated by numerical simulations for exposure to ELF electric and magnetic fields. Computations are based on the finite-difference time-domain method (FDTD). The computational uncertainty is determined by comparison of analytical and numerical results and amounts to a worst-case expanded uncertainty (95% confidence interval) of +/-9.89 dB for both dosimetric quantities (E, J). Detailed investigations based on the Visible Human body model with a resolution of 2 mm show a strong influence of the tissue boundaries on the simulation results, which is caused by the numerical method. For the tissue specific in situ electric field and current density changes in excess of 10 dB are observed when comparing the results with and without evaluation of the dosimetric quantities at tissue boundaries. Moderate sensitivities with respect to tissue boundaries are observed only for low conductivity tissues when evaluating the in situ electric field whereas this behavior is observed for high conductivity tissues when evaluating the current density. For exposure to a 50 Hz magnetic field corresponding to the ICNIRP reference level, the simulated current density for central nervous system (CNS) tissue is in compliance with the ICNIRP guidelines. Exposure to a 50 Hz electric field may exceed the ICNIRP basic restriction for CNS tissue at least in a worst-case scenario (grounded human body, vertical electric field, tissue boundaries included for the evaluation of the current density). The in situ electric field is the more stable dosimetric quantity with respect to changes of the tissue conductivity of the Visible Human body model. The maximum conductivity sensitivity coefficient amounts to +122% for the current density whereas the maximum sensitivity coefficient for the in situ electric field is -20%. For electric field exposure the in situ electric field remains comparable (-6% to -4%), the averaged current density change ranges from -57% to -16% for the tissues under investigation. Magnetic field exposure of a scaled model of a five year old child leads to a decrease of the dosimetric quantities (J: -74% to -45%, E: -42% to -23%) compared to the Visible Human results.

BAliBASE (Benchmark Alignment dataBASE): enhancements for repeats, transmembrane sequences and circular permutations.

BAliBASE is specifically designed to serve as an evaluation resource to address all the problems encountered when aligning complete sequences. The database contains high quality, manually constructed multiple sequence alignments together with detailed annotations. The alignments are all based on three-dimensional structural superpositions, with the exception of the transmembrane sequences. The first release provided sets of reference alignments dealing with the problems of high variability, unequal repartition and large N/C-terminal extensions and internal insertions. Here we describe version 2.0 of the database, which incorporates three new reference sets of alignments containing structural repeats, trans-membrane sequences and circular permutations to evaluate the accuracy of detection/prediction and alignment of these complex sequences. BAliBASE can be viewed at the web site http://www-igbmc.u-strasbg. fr/BioInfo/BAliBASE2/index.html or can be downloaded from ftp://ftp-igbmc.u-strasbg.fr/pub/BAliBASE2 /.

Results of plate fixation of type 5 olecranon fractures in 20 horses.

REASONS FOR PERFORMING STUDY: Previous olecranon fracture reports contain a small proportion of type 5 fractures, mostly treated with conservative therapy. OBJECTIVES: To evaluate the clinical details and outcome of type 5 olecranon fractures in a large group of horses treated by tension band plate fixation and to compare results with other treatment methods. METHODS: Medical records of 97 cases, including 32 (33%) classified as type 5, were reviewed. Subject details, history, radiographic findings, treatment and follow-up results (2-146 months post operatively) were recorded. RESULTS: Treatment included open reduction and internal fixation using a narrow or broad dynamic compression plate (n = 20), conservative therapy (n = 7) and euthanasia (n = 5). Long-term follow-up was available in 15 cases treated surgically, of which 2 were sound and in training, 11 sound and performing athletically and 2 unsound. Distal semilunar notch involvement, comminution or open status did not appear to affect prognosis. CONCLUSIONS: Internal plate fixation provides an excellent prognosis for an animal to be capable of athletic performance. POTENTIAL RELEVANCE: Describing tension band plate fixation and results offers a method of fracture repair that should improve treatment and prognosis for type 5 olecranon fractures.

Point mutations causing Bloom's syndrome abolish ATPase and DNA helicase activities of the BLM protein.

Bloom's syndrome (BS) is a rare human genetic disorder characterized by mutations within the BLM gene whose primary effects are excessive chromosome breakage and increased rates of sister chromatid interchange in somatic cells. We report the characterization of a murine protein (mBLM), highly related to the product of the human BLM gene. This protein exhibits an ATP-dependent DNA-helicase activity that unwinds DNA in a 3'-5' direction. Single amino acid substitutions found in BS cells, abolish both ATPase and helicase activities of this protein, indicating that defects in these BLM functions may be primarily responsible for BS establishment. These results provide the first evidence suggesting that the enzymatic activities of the BLM product are implicated in the upholding of genomic integrity.

A murine ATFa-associated factor with transcriptional repressing activity.

The ATFa proteins, which are members of the CREB/ATF family of transcription factors, have previously been shown to interact with the adenovirus E1a oncoprotein and to mediate its transcriptional activity; they heterodimerize with Jun, Fos or related transcription factors, possibly altering their DNA-binding specificity; they also stably bind JNK2, a stress-induced protein kinase. Here we report the identification and characterization of a novel protein isolated in a yeast two-hybrid screen using the N-terminal half of ATFa as a bait. This 1306-residue protein (mAM, for mouse ATFa-associated Modulator) is rather acidic (pHi 4.5) and contains high proportions of Ser/Thr (21%) and Pro (11%) residues. It colocalizes and interacts with ATFa in mammalian cells, contains a bipartite nuclear localization signal and possesses an ATPase activity. Transfection experiments show that mAM is able to downregulate transcriptional activity, in an ATPase-independent manner. Our results indicate that mAM interacts with several components of the basal transcription machinery (TFIIE and TFIIH), including RNAPII itself. Together, these findings suggest that mAM may be involved in the fine-tuning of ATFa-regulated gene expression, by interfering with the assembly or stability of specific preinitiation transcription complexes.

In vivo association of ATFa with JNK/SAP kinase activities.

The human ATFa proteins belong to the CREB/ATF family of transcription factors. We have previously shown that the ATFa proteins may contribute to the modulation of the transcriptional activity of the Jun/Fos complexes (Chatton et al. (1994). Oncogene, 9, 375-385). We now show that a protein kinase activity is strongly associated with ATFa in vivo, as revealed by coimmunoprecipitation of ATFa/kinase complexes from whole cell extracts, with antibodies against ATFa. Two independent regions were found to be implicated in kinase binding: a major interaction site is located within the N-terminal 82 residues comprising an important metal-chelating element; a weaker binding site corresponds to the basic sequence element preceding the C-terminal leucine-zipper of ATFa. Induction experiments suggest that each of these ATFa domains may interact with different kinases. The major activity is associated with the ATFa N-terminal domain. Based on its response to various inducers, on both in vitro and in vivo binding assays, and on its immunological properties, this activity most likely corresponds to the 54/55 kDa JNK2 protein. Taken together, these observations suggest that the ATFa proteins, among other CREB/ATF proteins, may be important effectors of cell signalling pathways.

Role of the ATFa/JNK2 complex in Jun activation.

The ATFa proteins, which are members of the CREB/ATF family of transcription factors, display quite versatile properties. We have previously shown that they interact with the adenovirus E1a oncoprotein, mediating part of its transcriptional activity and heterodimerize with the Jun, Fos or related transcription factors, thereby modulating their DNA-binding specificity. In the present study, we report the sequence requirement of the N-terminal activation domain of ATFa and demonstrate the importance of specific threonine residues (Thr51 and Thr53) in addition to that of the metal-binding domain, in transcriptional activation processes. We also show that the N-terminal domain of ATFa which stably binds the Jun N-terminal kinase-2 (JNK2) (Bocco et al., 1996), is not a substrate for this kinase in vivo but, instead, serves as a JNK2-docking site for ATFa-associated partners like JunD, allowing them to be phosphorylated by the bound kinase.

Eukaryotic GST fusion vector for the study of protein-protein associations in vivo: application to interaction of ATFa with Jun and Fos.

We describe a multipurpose eukaryotic expression vector that incorporates the following features: restriction sites for in-frame insertion of cDNAs of interest between sequences encoding the glutathione-S-transferase (GST) and an oligohistidine element, allowing expression of the corresponding fusion proteins; a phosphorylation site for protein kinase A for in vitro labeling of the fusion protein; a T7 promoter for in vitro transcription and subsequent translation; and signals for single-stranded DNA production in bacteria. We have used this vector to demonstrate the formation in vivo of complexes between the transcription factor ATFa, a member of the family of ATF/CRE binding proteins, and the c-Jun or c-Fos proteins. Such interactions could be detected in crude extracts from cells transfected with vectors expressing the GST-ATFa fusion protein, as well as the c-Jun or c-Fos proteins. Complexes containing both ATFa and either c-Jun or c-Fos were specifically retained on glutathione (GSH)-agarose beads as revealed by immunoblot analyses. We also show that the leucine zipper domain of ATFa is essential for this interaction.

Hepatic extraction efficiency and excretion rate of technetium-99m-mebrofenin in dogs.

UNLABELLED: Quantitative hepatobiliary scintigraphy aids in the diagnosis of hepatic disease. Two scintigraphic parameters that have great value in discriminating between hepatocellular and biliary disease are hepatic extraction fraction (HEF), which is a measure of the hepatic extraction efficiency (HEE), and hepatic excretion rate. It is generally accepted that hepatic extraction fraction is normally 100%, but a review of the literature provided little information on the actual HEF of 99mTc-mebrofenin. METHODS: We determined the HEE of 99mTc-mebrofenin in nine normal dogs after direct injection into the afferent hepatic vasculature using a two-compartment model. The forward and reverse rate constants for the two-compartment model were solved by a simple graphic approach and a more complex numerical approach using a nonlinear least squares algorthm. The HEEs were determined using both methods. RESULTS: The HEE for the graphic and numerical methods of analysis were not significantly different and were calculated to be 92.2 +/- 4.75% (mean +/- s.d.) and 91.2 +/- 4.44% (mean +/- s.d.) by each method, respectively. The half-time clearance of 99mTc-mebrofenin was 19.10 +/- 4.86 min (mean +/- s.d.). CONCLUSION: This study validates the assumption that the normal HEE of 99mTc-mebrofenin is nearly 100%, barring species differences.

Lipospermine-mediated gene transfer technique into murine cultured cortical cells.

In order to transfer exogenous DNA into embryonic cortical cells, we have chosen a transfection technique using a synthetic lipospermine (dipalmitoylphosphatidylethanolamylspermine, DPPES) which complexes DNA molecules and allows their penetration into the intracellular compartment. The procedure was optimized after testing several parameters: DPPES/DNA ratio, incubation time, kinetics of transgene expression, and growth medium. The protocol was achieved by following the expression of the E. coli LacZ reporter gene under the control of the cytomegalovirus promoter. The lipopolyamine-mediated transfection is efficient for terminally differentiated cells, since we routinely obtained transfection efficiencies of 30% for neurons.

Cerebral arterial spasm: part 10. Reversal of acute and chronic spasm in dogs with orally administered nifedipine.

In vivo experiments in dogs demonstrated angiographically that the subarachnoid injection of blood produced cerebral arterial apasm both immediately after the injection of blood and 2 days later. The sublingual adminstration of nifedipine reversed both the acute and the delayed cerebral arterial spasm. In addition, sublingual administration of nifedipine 20 minutes before the subarachnoid injection of blood prevented the acute spasm.

Percutaneous catheter drainage in intraabdominal fluid collections including infected biliary ducts and gallbladders.

Sixty-five abscesses, including 6 infected biliary systems and 15 sterile fluid collections, were treated by percutaneous catheter drainage in 77 febrile patients who were evaluated by computerized tomography or ultrasonography of intraabdominal infection. Percutaneous catheter drainage and systemic antibiotic administration without surgery provided satisfactory control of infection in 52 of 65 abscesses (80 percent). Catheter drainage followed by surgical exploration for abscess control was performed in an additional 5 of 65 abscesses (7 percent). Nine death (14 percent) occurred in the abscess group of 64 patients. In 15 patients, aspirations, Gram stain, and culture of the abnormal fluid collection revealed sterile fluid. Drainage with a single catheter allowed complete resolution in 14 of 15 sterile collections. Surgery was performed electively in one patient with a fistula from a pancreatic pseudocyst in the small bowel. No deaths occurred in the noninfected group of 15 patients, 2 of whom underwent drainage of coexisting abscesses.