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PubMed1994

Acquisition and use of calorimetric data for prediction of the thermodynamics of ligand-binding and folding reactions of proteins.

The commercialization of high-precision microcalorimeters has led to a sharp increase in the acquisition and utilization of calorimetric data on the binding and folding reactions of proteins. This thermodynamic database, together with high-resolution structures of proteins and their complexes with ligands, has facilitated the development, calibration, refinement and testing of empirically based and computational methods for predicting the thermodynamics of processes involving macromolecules. Even so, a generally accepted method for assigning structural parameters to the various energetic contributions of protein folding and ligand-binding reactions, much less a method for predicting the changes in thermodynamic parameters of these reactions, has not yet been firmly established.

Calorimetry↗
PubMed2005

Analysis of nitrogen fractions in silage by near-infrared spectroscopy.

Total nitrogen, soluble nitrogen (SN), nonprotein nitrogen (NPN), and acid-detergent insoluble nitrogen (ADIN) were analyzed in grass silage by near-infrared (NIR) spectroscopy. A set of 144 samples was used to calibrate the instrument by modified partial least-squares regression, and the following statistical results were achieved: standard error of calibration (SEC) = 0.449 and square correlation coefficient (R (2)) = 0.98 for total nitrogen x 6.25, SEC = 0.425 and R (2) = 0.95 for SN x 6.25, SEC = 0.414 and R (2) = 0.94 for NPN x 6.25, and SEC = 0.139 and R (2) = 0.84 for ADIN x 6.25. To validate the calibration performed, a set of 48 silage samples was used. Standard errors of prediction were 0.76, 0.64, 0.63, and 0.25 for total nitrogen, SN, NPN, and ADIN (all of them multiplied by 6.25), respectively, and R (2) for the regression of measurements by reference method versus NIR analysis were 0.94, 0.92, 0.90, and 0.48 for total nitrogen, SN, NPN, and ADIN, respectively. To compare the results obtained by NIR spectroscopy with those obtained by the reference methods for total nitrogen, SN, and NPN of the validation set, linear regression and paired t tests were applied, and the results were not significantly different (p = 0.05). When mean square prediction error analysis was applied, it could be concluded that for total nitrogen, SN, and NPN, a robust calibration model was obtained and that the main error was unexplained error. Statistical data for ADIN were worse than those of the other parameters; as a result NIR spectroscopy is not an effective method for quantitative analyses of ADIN in silage; nevertheless, it may be an acceptable method for semiquantitative evaluation.

Calibration↗
PubMed1965

METHOD FOR THE AUTOMATIC DETERMINATION OF SERUM IRON.

A simple method is described for the automated determination of serum iron and tota iron-binding capacity, which eliminates the need to render apparatus or reagents iron-free. The method is faster than normal procedures and its accuracy is comparable with other accepted methods for the determination of serum iron.

Blood Chemical Analysis↗
PubMed1998

Efficacy of sterilisation methods and their effect on enamel demineralisation.

The aim of this project was to determine the effectiveness of sterilisation methods for dental enamel for use in intra-oral cariogenicity tests, and their possible effect on the degree of demineralisation of enamel. Bovine incisors were cut vertically into five portions and each assigned to one of five groups. Group 1 was used as a control while the other four groups were subjected, respectively, to gamma irradiation ( congruent with 25kGy), steam autoclaving (121 degrees C for 15 min), sodium hypochlorite (12% w/v for 24h) and povidone-iodine (7.5% w/v for 24h). Total viable counts of microorganisms remaining following sterilisation of the specimens were performed following incubation of the specimens for 24h at 37 degrees C. Caries-like lesions were produced in each specimen using an acidic buffer solution (pH4.5). Sections were cut from each specimen, ground to 80-microgram thickness, and microradiographed. Mineral loss and lesion depth were quantified using transverse microradiography. Statistical analysis was by ANOVA. Dunnett's and Tukey's tests. Microbial growth (Staphylococcus aureus and bacilli) was observed only in control specimens in both brain heart infusion broth and on blood agar plates. The sterilisation methods affected the enamel surface as follows: gamma irradiation (cream discolouration), NaOCl (bleaching), and povidone-iodine (white spot-like lesion). Compared with the control, there was no significant difference in mineral loss and lesion depth with any of the groups, but the numerical values of mineral loss and lesion depth in groups can be ranked as follows: gamma irradiation <povidone-iodine <control <autoclave <NaOCl. In conclusion, the four sterilisation methods were all effective to sterilise enamel, but gamma irradiation proved the most acceptable method for enamel to be used in cariogenicity tests having the least adverse effect.

Acids↗
PubMedDate not supplied

Treatment of laryngeal stenosis. A conservative new approach.

One of the most perplexing problems in laryngology is the treatment of laryngeal scar tissue. This can be supraglottic, glottic, or infraglottic. Two acceptable methods of treatment have been surgical and endoscopic. Endoscopic treatment consists of dilatation, excision of scar tissue with a laser, removal of scar tissue with insertion of a stent, and removal and scar tissue with injection of intralesional cortisone. Twelve cases are reported, treated with microscopic removal of scar tissue followed by voice rest and beclomethasone dipropionate (Vanceril) inhalations, a topical steroid. All twelve cases showed marked improvement measured by improved glottic opening and improvement of voice quality. Further evaluation of this method of treatment should be done as an auxiliary treatment to our present accepted regime of treatments in this troublesome condition.

Adult↗
PubMedDate not supplied

Epithelial ovarian cancer: prevention, diagnosis, and treatment.

The leading cause of death from gynecologic malignancies in the United States is epithelial ovarian cancer. The significant risk factor for development of ovarian cancer is advancing age, although there is clearly a genetic predisposition--often associated with the BRCA1 and BRCA2 genes--in at least 5% to 10% of all epithelial ovarian cancers. Oral contraceptives are known to reduce the risk for development of ovarian cancer and should be considered as a method of birth control in women at increased risk. Currently, there is no acceptable method of screening for this disease, although measurement of CA-125 level and transvaginal ultrasound have been utilized. Ovarian cancer is a surgically staged disease. In apparent early-stage disease, complete surgical staging is critical for the selection of adjunctive therapy. In advanced-stage disease, the goal is primary cytoreduction. Standard postoperative therapy for advanced-stage ovarian cancer includes platinum-based chemotherapy with the substitution of paclitaxel for cyclophosphamide occurring in the last decade. Despite these advances in chemotherapy, ovarian cancer continues to be fatal in far too many cases.

Carcinoma↗
PubMedDate not supplied

Iatrogenic ulnar nerve injury after percutaneous cross-pinning of supracondylar fracture in a child.

Supracondylar fracture of the humerus is the most common fracture of the elbow in children and has been treated by a variety of methods. Recently, stabilization of reduced fractures with percutaneous pin fixation has become the accepted method of treatment. Ulnar nerve injury is a complication of percutaneous pinning of supracondylar fractures, although many authors have reported that it resolves spontaneously after removal of the pin.

Bone Nails↗
PubMedDate not supplied

[Determination of the degree of severity of cerebral contusions].

Three degrees of severity of the brain contusion are generally distinguished today for working out the most rational and purposeful treatment. It is quite difficult to judge the severity of the contusion from the neurological picture alone because neurological manifestations of the brain contusions are very diverse and dynamic in character. The authors analysed 630 case records of patients with contusion of the brain and disclosed signs most characteristic of each degree of contusion. In appraising the severity of the contusion, the authors recommend resorting to the laboratory and instrumental diagnostic methods accepted currently in neurosurgical practice in addition to the neurological examination. The complex method of appraising the patients' condition makes the determination of the degree of severity of the brain contusion more trustworthy and objective.

Brain Concussion↗
PubMed1996

Consensus minimum standards for use in a trainer's report for summative assessment in general practice.

BACKGROUND: Summative assessment of general practitioner registrars is to be introduced in September 1996, one component of which will be a report by the trainer. Standards must be set and guidance provided as to the most appropriate ways of obtaining evidence for the trainer's assessment. AIM: The first aim of this study was to set consensus minimum standards for 30 items that are likely to form the content of a trainer's report; the second aim was to provide a consensus view on the most appropriate methods of assessment to be used by trainers. METHOD: A consensus conference was held in March 1995 during which the 30 items were discussed by a group of 30 general practitioners, of whom 27 were experienced trainers. This resulted in a draft document that was circulated to the conference attenders and other experts for consultation. RESULTS: Draft minimum standards were produced for all 30 items after the consensus conference with a mean of 2.5 standards for each item. Of those involved in the consultation exercise, 82% replied. Most of the revisions suggested at this stage were of a minor nature; the only major revision was to divide one item into two, resulting in a final total of 31 items. All but one of the 80 standards could be assessed by direct observation; 41 (51%) could be assessed by tutorial-based discussion and 61 (76%) by methods specific to that standard. Trainers or their practice partners were viewed as acceptable sources of evidence for all items and hospital consultants and primary health care team members were viewed as acceptable for just over half of the items. CONCLUSION: Standards for use by trainers when providing a general practitioner report for the summative assessment of registrars have been developed by consensus conference and have been subjected to review by consultation. Acceptable methods by which registrars could be assessed against these standards, and suitable personnel who could provide evidence, have also been suggested.

Education, Medical, Graduate↗
PubMed1994

Improved method for analysis of whole bone torsion tests.

Structural tests, such as whole bone torsion tests, have become widely accepted methods for assessing average bone material properties. To simplify interpretation of these tests, the nonuniform bone geometry is often analyzed as a tube with a constant cross section (prismatic) and the areal properties of the smallest bone section. This approach may not adequately represent the true torsional behavior of the cross section and does not account for any lengthwise variations in bone geometry. The errors introduced by these approximations are particularly significant when comparing bones of different sizes and geometries. In this paper, we examine the effects of approximating the cross-sectional torsional behavior and of neglecting lengthwise variations in bone geometry. We then present a simple, standardized procedure utilizing a FORTRAN computer program for accurate determination of material properties. We examine first simple idealized bone geometries and then a complex three-dimensional model of the femur from a 26-day-old male Sprague-Dawley rat. For these models, the conventional methods for interpreting torsion tests introduce errors of up to 42% in the shear modulus and up to 48% in the maximum shear stress; a straightforward extension of these methods reduces the errors to within 3%.

Animals↗
PubMed1995

Application issues in bioanalytical method validation, sample analysis and data reporting.

Although some degree of consensus has been reached concerning the requirements for acceptable method validation, the procedures used to establish them vary significantly between laboratories. Also, issues arising from application of these requirements during validation and subsequent sample analysis need to be addressed. The purpose of this paper is to discuss application issues concerning prerequisites to method validation, and all validation criteria for evaluation of method reliability and overall performance. Other poorly addressed issues such as re-validation, cross-validation, partial sample volume, multicomponent analysis and reporting will also be discussed. Although many issues discussed are of a general nature, the scope of this presentation is primarily to address issues arising from the validation and routine application of chromatographic methods.

Animals↗
PubMed2002

Enhancement of combinatorial chemistry by microwave-assisted organic synthesis.

It was in the 1980 s that the first papers in which the use of either combinatorial methods or microwave heating in organic chemistry were published. Unlike combinatorial chemistry, which quite readily became an accepted method, particularly in the pharmaceutical industry, it is only now that microwave heating is truly gaining acceptance. Our aim in this review is to attempt to rationalize this slow acceptance and to show the benefits to be gained by employing microwave heating in tandem with combinatorial chemistry. We will also give a number of examples of successful applications.

Chemistry, Organic↗
PubMed1983

Migration of human leukocytes from soft agarose droplet: a simplified method for studying chemotaxis and spontaneous migration.

Studies of in vitro chemotaxis and spontaneous migration of human leukocytes using the accepted method with the Boyden-chamber-filter are troublesome, because of the need for specially constructed vessels as well as the difficulties caused by the use of membrane filters. We describe a new and simplified method for measuring human leucocyte chemotaxis, which is a modification of the recently described underagarose migration method and which is based upon spontaneous migration of cells from a soft agarose droplet and in response to a chemotactic gradient. We examined suspensions of leukocytes, purified granulocytes, and mononuclear cells from 10 healthy normal adults and from 10 samples of cord blood using E Coli O111B4 endotoxin-activated human serum as attractant. Our results showed that the mean chemotactic indices (C.I.-chemotaxis/migration) for purified granulocytes and for mononuclear cells from normal individuals were 3.0 +/- 1.2 and 2.7 +/- 1.5, respectively. Chemotaxis was significantly reduced when unwashed leukocytes were studied, indicating a detrimental effect of autologous plasma on leukocytic response to a chemotactic stimulus in this system. Cord blood cells showed normal spontaneous migration, but significantly decreased chemotaxis. This preliminary report shows that the technique is simple, rapid, and reproducible, and can detect abnormalities of chemotaxis in both granulocytes and mononuclear cells.

Adult↗
PubMed1992

Laparoscopic varicocelectomy: preliminary report of a new technique.

The use of varicocelectomy for the treatment of subfertility seems to be incontrovertible. However, there is a difference of opinion as to the proper surgical method of varicocele ablation. The inguinal and high retroperitoneal approaches are the most commonly accepted methods to date. However, significant postoperative morbidity is common and return to normal activity often is prolonged. Also, bilateral operations are being performed more commonly. These considerations have prompted many to search for alternative techniques. We developed a laparoscopic procedure that is as simple and effective as more traditional methods. In addition, it offers lower morbidity, allows for microscopic dissection with preservation of the spermatic artery and is amenable to bilateral ligation without a second incision. Ten patients 16 to 54 years old underwent laparoscopic ligation of the spermatic veins at the internal inguinal ring. The diagnosis was based on physical examination. Indications for the operation were infertility with a stress sperm pattern in 5 patients, testicular atrophy in 4 and scrotal pain in 1. Four patients underwent bilateral ligation. Preliminary followup showed resolution of the varicocele in all patients and disappearance of pain in the patient treated for this symptom. No morbidity related to this procedure has been encountered and all patients resumed normal activity within 2 days. We believe that this new method is a viable alternative for varicocelectomy.

Adolescent↗
PubMed1999

Effect of sterilisation methods on the structural integrity of artificial enamel caries for intra-oral cariogenicity tests.

Enamel blocks bearing artificial caries are used in intra-oral appliances for cariogenicity tests. These blocks are often sterilised to prevent the possibility of cross-infection via this route. This study therefore aimed to determine the effect of sterilisation methods on the structural integrity of artificial enamel caries used for intra-oral cariogenicity tests. Four experimental groups were devised. Ten bovine incisors were used in each group. Artificial caries was produced in each tooth which was subsequently cut into two halves. One half of each tooth was reserved as control while the other was sterilised. The four groups were subjected to respective sterilisation methods: gamma irradiation (approximately equal to 25 KGy), steam autoclaving (121 degrees C for 15 min), sodium hypochlorite (12% w/v for 24 h) and povidone-iodine (7.5% w/v for 24 h). The control and sterilised specimens in each group were examined for microbial growth after incubation in nutrient broth for up to 7 days at 37 degrees C under aerobic and anaerobic conditions. Mineral loss and lesion depth were quantified from microradiographs of sections from control and sterilised specimens using transverse microradiography. Data were analysed statistically by paired Student's t-test. Microbial growth was observed only in control specimens. Gamma irradiation and NaOCL caused cream discolouration and bleaching of the enamel surface, respectively. Autoclaving, sodium hypochlorite and povidone-iodine resulted in further demineralisation of the lesions. The four sterilisation methods were all effective sterilants for artificial caries. However, gamma irradiation appears the most acceptable method considering the more adverse effects of the other methods with regards to cariogenicity tests.

Animals↗
PubMed2003

Induced sputum in the investigation of airway inflammation of COPD.

During the last decade, the method of sputum induction (SI) has offered the opportunity to study inflammation in patients with chronic obstructive pulmonary disease (COPD). This paper reviews methodological aspects of SI and summarizes its uses in the research of inflammation in COPD, including sputum cellularity and soluble markers. SI is a relatively safe, reliable, and reproducible technique, used to investigate different aspects of airway inflammation. Although various methods of induction and processing have been proved safe and highly reproducible, a generally accepted method is needed. Sputum analysis has given evidence for increased numbers of macrophages and neutrophils in COPD patients compared to normal subjects. In some studies, increased numbers of eosinophils have been also reported. Changes in various mediators have been found in sputum supernatant of COPD patients (IL-8, LTB-4 and TNF-a). The clinical usefulness of the method in the follow-up of the disease has not been explored extensively. A number of observations in patients with different clinical characteristics could be proven useful in identifying patterns of inflammation associated with different prognosis. Finally, SI could also guide treatment; such as, sputum eosinophilia in COPD could predict response to inhaled corticosteroids.

Biopsy↗
PubMed2002

Validation of a method for the analysis of iron and manganese in table olives by flame atomic absorption spectrometry.

There is no generally accepted method for determination of the amounts of iron and manganese in table olives. Application of flame atomic absorption spectrometry to the analysis of both elements has been examined to validate a method that may be used by the industry's quality control laboratory as well as by the laboratories of regulatory agencies. The method has detection limits of 0.106 and 0.022 mg/L and quantification limits of 0.271 and 0.057 mg/L, for Fe and Mn, respectively, referred to the solution to be measured. There was no significant effect due to the matrix, but a slight bias due to the presence of Ca has been detected. Recoveries were excellent, and the method was robust. Influence of operator, HCl and Mg salt compounds, calcination equipment, or dates on results was not found. Relative errors were, in general, below 4% for both cations, and repeatability was below 3.43 and 0.38 mg/kg of olive paste for Fe and Mn, respectively. The method is proposed for the analysis of Fe and Mn in ripe olives and table olives in general.

Calibration↗
PubMed1981

Comparison of minimum inhibitory concentration values determined by three antimicrobic dilution methods for Pseudomonas aeruginosa.

This investigation compares minimum inhibitory concentration measurements by three antimicrobic dilution methods for Pseudomonas aeruginosa versus seven antimicrobics. Minimum inhibitory concentrations were measured for 650 P. aeruginosa clinical isolates and for repeated tests with P. aeruginosa ATCC 27853 versus gentamicin, tobramycin, amikacin, netilmicin, sisomicin, carbenicillin, and ticarcillin, using the macro-broth, micro-broth, and agar dilution methods. For all antimicrobics, it was found that the micro-broth and agar dilution methods produced comparable minimum inhibitory concentration measurements, which were found to lie 1 to 2 double dilution steps below those determined by the macro-broth method. Acceptably replicability was found for both the macro-broth and the agar dilution methods. The micro-broth method showed less replicability, with 4.7% of minimum inhibitory concentration values lying +/- 2 or more double dilution steps from the modal value. It is important to recognize such differences if micro-broth or agar dilution methods are substituted for the macro-broth method.

Aminoglycosides↗