Single versus multiple metabolite quantification of in vitro starch digestion: A comparison for the case of pulse cotyledon cells.

Different quantification methods for in vitro amylolysis were compared for individual chickpea and lentil cotyledon cells (ICC) as a relevant case study. For the first time, much-applied spectrophotometric methods relying on the quantification of certain functional groups (i.e., DNS, GOPOD) were compared to chromatographic quantification of starch metabolites (HPLC-ELSD). The estimated rate constant and linked initial rates of amylolysis were highly correlated for DNS, GOPOD, and HPLC-ELSD. However, absolute amylolysis levels depended on the applied method and sample-specific metabolite formation patterns. Multiresponse modelling was employed to further investigate HPLC-ELSD metabolite formation patterns. This delivered insight into the relative importance of different amylolysis reactions during in vitro digestion of pulse ICC, proving that maltotriose and maltose formation determined the overall amylolysis rate in this case. Multiresponse reaction rate constants of maltotriose and maltose formation were highly correlated to single response amylolysis rate constants (and initial rates) obtained for all three quantification methods.

The Value of Intraoperative 3-Dimensional Fluoroscopy in the Treatment of Distal Radius Fractures: A Randomized Clinical Trial.

PURPOSE: This study attempted to determine the clinical effectiveness of the intraoperative use of 3-dimensional fluoroscopy compared with conventional 2-dimensional fluoroscopy in patients with distal radius fractures. METHODS: We performed a multicenter randomized clinical trial in which 206 patients were randomized between the use of 3-dimensional fluoroscopy or not during operative treatment of the distal radius fracture. The primary outcome was the quality of fracture reduction and fixation assessed on a postoperative computed tomography scan with a dichotomous outcome: indication for revision, yes or no. RESULTS: There was no significant difference in whether the fracture required revision surgery: 31% (2-dimensional group) versus 24% (3-dimensional group). In 11% of distal radius fractures allocated to the 3-dimensional group, additional intraoperative corrections (screw replacements) were performed. CONCLUSIONS: Compared with 2-dimensional fluoroscopy, the use of intraoperative 3-dimensional fluoroscopy does not appear to improve the quality of reduction and fixation in the management of patients with a distal radius fracture. However, the use of 3-dimensional fluoroscopy appears to have advantages such as more intraoperative revisions and less revision surgeries that this study could not clearly demonstrate. TYPE OF STUDY/LEVEL OF EVIDENCE: Diagnostic II.

Emerging drugs and alternative possibilities in the treatment of tuberculosis.

INTRODUCTION: Tuberculosis (TB) remains a global health problem. Drug resistance, treatment duration, complexity, and adverse drug reactions associated with anti-TB regimens are associated with treatment failure, prolonged infectiousness and relapse. With the current set of anti-TB drugs the goal to end TB has not been met. New drugs and new treatment regimens are needed to eradicate TB. AREAS COVERED: Literature was explored to select publications on drugs currently in phase II and phase III trials. These include new chemical entities, immunotherapy, established drugs in new treatment regimens and vaccines for the prophylaxis of TB. EXPERT OPINION: Well designed trials, with detailed pharmacokinetic/pharmacodynamic analysis, in which information on drug exposure and drug susceptibility of the entire anti-TB regimen is included, in combination with long-term follow-up will provide relevant data to optimize TB treatment. The new multi arm multistage trial design could be used to test new combinations of compounds, immunotherapy and therapeutic vaccines. This new approach will both reduce the number of patients exposed to inferior treatment and the financial burden. Moreover, it will speed up drug evaluation. Considering the investments involved in development of new drugs it is worthwhile to thoroughly investigate existing, non-TB drugs in new regimens.

Internal fixation of fractures of the proximal humerus with the MultiLoc nail.

OBJECTIVE: Anatomical reduction of two- to four-part fractures of the proximal humerus using indirect reduction techniques. Intramedullary fixation with a short humerus nail. Restoration of a stable bone-implant construct that enables early functional after-treatment. INDICATIONS: Displaced and unstable two- to four-part fractures of the proximal humerus. Fractures of the proximal humerus extending in the humeral diaphysis (use a long nail). Ipsilateral combined lesions of the proximal humerus and the humeral diaphysis (use a long nail). CONTRAINDICATIONS: Poor physical and/or mental status. Critical soft tissue conditions in the area near the surgical site. Local soft tissue infection. Pre-existing severe osteoarthritis of the shoulder joint; severe shoulder stiffness. Head-split fractures of the humerus head that cannot be reduced. SURGICAL TECHNIQUE: Exposure of the fracture using an anterior acromial approach and determination of the correct nail entrance point. Anatomic fracture reduction using indirect reduction techniques. Stable fixation using an intramedullary MultiLoc® nail. Determination of the proximal locking configuration depending on the fracture morphology. Distal locking with angle-stable option. POST-OPERATIVE TREATMENT: Post-operative radiographs for documentation of the surgical result and implant position. Use of an arm sling for 7-10 days. Active and passive exercises of the shoulder joint starting on day 1. Shoulder abduction limited to 60° for 2 weeks. Subsequent abduction to 90° until the 4th week. Subsequent active mobilisation without restrictions. Weight bearing and sporting activities after 3 months. Radiological evaluation after 2, 6 and 12 weeks. RESULTS: During a 6-month period, 160 patients were documented in a prospective clinical multicentre study. According to the AO classification, there were 36% A-type fractures, 41% B- and 23% C-type injuries. A 6-month follow-up was available for 17 patients. The mean age of these patients was 67 years. One patient had an A-type fracture. There were ten B- and six C-type fractures. At the time of follow-up, the mean Constant score was 66 points. Radiographically, all fractures had healed. Intra-articular screw penetration and loss of reduction were both observed once.

A prospective cohort study comparing the VAS spine score and Roland-Morris disability questionnaire in patients with a type A traumatic thoracolumbar spinal fracture.

The Roland Morris Disability Questionnaire (RMDQ-24) and the VAS spine score have been regularly used to measure functional outcome in patients with back pain. The RMDQ-24 is primarily used in degenerative disease of the spine and the VAS Spine is used in trauma patients. The aim of this study is to compare these scores and to see if there is a correlation in patients with a traumatic thoracolumbar spinal fracture. Prospective cohort study comparing the RMDQ-24 and the VAS spine score in patients with a traumatic type A fracture thoracolumbar spine fracture. Fifteen non-operatively patients (group one) completed 118 questionnaires and 17 operatively treated patients (group two) completed 140 questionnaires. Group one scored an average of 6.6 and 65.9 for the RMDQ-24 and VAS Spine, in group two this was 5.1 and 82.9. Spearman's correlation test showed a significant correlation, in group one 0.83 and for the second group 0.87. RMDQ-24 and VAS Spine have a strong positive correlation in measuring disability in a group of patients with back pain because of a spinal fracture. In both non-operatively and operatively treated groups this correlation is significant.

Early proximal row carpectomy after severe carpal trauma.

Complex fracture dislocations of the wrist often result in post-traumatic arthrosis. In these cases, patients can experience severe pain and loss of function of the wrist and as a result many of them end up having a total arthrodesis. In trying to avoid the disadvantages of a total arthrodesis, alternative treatment strategies have been investigated, amongst which proximal row carpectomy (PRC). Basic conditions for a good outcome of PRC are an intact cartilage of the lunate fossa of the distal radius and an intact surface of the head of the capitate for the new radiocapitate joint (, schematic drawing post-PRC). Also, an intact (volar) radioscaphocapitate (RSC) ligament is necessary because it plays an essential role in stabilizing the new joint and preventing volar dislocation and ulnar translocation of the distal carpal row. Acute post-traumatic PRC can be indicated, but is rarely reported in literature. In this article, we present four patients whom we have treated with early PRC after severe trauma of the wrist. Three patients had a good outcome. In the patient with the bad outcome, the before-mentioned prerequisites were not met, which is discussed.

Model of good practice tools for risk reduction and clinical governance.

Information and Communication Technologies (ICT) are expected to support Healthcare Professionals in reducing medical errors, making the most relevant decisions and finding the most appropriate procedure for each patient. In particular, Knowledge Management and Decision Support Systems provide access to high quality information and to appropriate protocols. The present paper aims at comparing the approaches used in three ongoing R&D projects in order to support risk reduction and clinical governance. This comparison will lead to the presentation of a generic model of Decision Support Tools that transform shared and documented "Good Practices" into software entities that can pro-actively advice users in their daily work or when they encounter difficult situations.

Hypothermia in trauma patients.

Accidental hypothermia (AH) can be defined as an unintentional decrease in core temperature below 35 degrees C during cold exposure by individuals without intrinsic thermoregulatory dysfunction. Pathophysiological changes can be attributed both to the severity of hypothermia and to co-morbid factors such as trauma, submersion, intoxication and underlying diseases. In trauma victims stratified according to the Injury Severity Score, the factor hypothermia is considered to be a poor prognostic sign for survival. In these patients rewarming therapy should be applied as soon as possible. In the Utrecht University Hospital, adult patients with AH are managed according to an algorithm based on their presenting hemodynamic conditions. Patients with perfusing cardiac rhythms and systolic pressures over 80 mmHg will receive continuous arteriovenous rewarming (CAVR). Arrested and hemodynamically instable patients are treated with cardiopulmonary bypass (CPB). In a 3-year period, 22 patients with AH were admitted to the emergency department. Fourteen patients had a trauma as the cause of hypothermia. Twenty patients were treated according to the algorithm (CAVR n = 18, CPB n = 2) and two patients were alternatively managed. Mortality in the CAVR group was 28% and total mortality reached 32%. Complications of CAVR are related to placement and removal of vascular catheters and may be severe in these patients with impaired coagulation. CAVR permits a good access to the (trauma-) patient and rewarming can be continued synchronously with diagnosis and treatment of various injuries.

Quantitation of IgG and IgM human anti-mouse antibodies (HAMA) interference in CA 125 measurements using affinity chromatography.

Currently no available immunoassay system offers complete protection against spuriously elevated or lowered results due to interference by Human Anti-Mouse Antibodies (HAMA). Although routine use of chromatography procedures is not an acceptable option because of the extra cost and workload involved, such a procedure would be highly desirable to ensure accurate immunoassay results. The present report describes a relatively simple affinity chromatography procedure using a HiTrap Protein G column to isolate immunoglobulin G (IgG) HAMA, followed by a HiTrap N-hydroxy-succinimide (NHS)-activated column coupled to goat-anti human immunoglobulin M (IgM) to bind IgM HAMA. To examine the usefulness of this purification procedure we determined CA 125 in forty serum samples prior to and following chromatography. Pre- and post-injection samples were obtained from 20 patients injected with 1 mg of 111In-labelled murine OC 125 F(ab')2 fragments in an immunoscintigraphy study. It is shown that this analytical procedure provides a technique to determine the extent and the nature of the existing HAMA interference in samples of patients after in vivo use of monoclonal antibodies for diagnostic or therapeutic purposes. The procedure can also contribute to the clarification of clinically discordant CA 125 results. Finally, the availability of such a procedure in the clinical laboratory provides an opportunity to test the robustness of newly developed immunoassay systems towards HAMA interference.

Roe v. Wade. Reality check.

PIP: For US Catholics, abortion, as predicted in 1968 by Robert Drinan, has become a public policy question that places the church in the position of "sinning by the use of its prestige and power against the sincerely held convictions of non-Catholics and non-believers, or as a group which failed by silence to speak up when misguided men and women changed the law to permit the extermination of undesirable and unwanted human beings." During the 25 years since the Roe vs. Wade decision, Catholics for a Free Choice has made a sustained effort to respond to this dilemma by providing an alternative view within the church and larger society. Abortion should remain legal to protect the desperate women who would otherwise resort to unsafe abortion. Also, the tradition of Catholic legal realism dating from Thomas Aquinas notes that good lawmaking requires the possibility of compliance and enforceability. If antiabortion laws are not enforced, are not obeyed, and are subject to widespread noncompliance, they are not sound law. Because the morality of abortion remains questionable, society should use noncoercive measures to reduce its incidence. Catholic moral teaching and Catholic views on social justice could make a positive contribution in this regard. Abortion has been a central issue for Roman Catholic political participation in the past 25 years as the National Conference of Catholic Bishops has attempted to reverse the Roe decision. During this time, Catholics for a Free Choice has provided an important alternative forum.^ieng

Comparison of four 'second generation' immunoassay systems to determine CA 125 in serum by using a graphical approach to method comparison analysis.

Clinical management of ovarian cancer patients is facilitated by CA 125 determinations in serum. Presently, several assay systems based on different concepts and different methodologies are available to measure CA 125. Method comparison analysis of such assay systems is usually performed through (linear) regression analysis, which requires assumptions about the distribution of experimental data and its measurement error. The aim of the present study was to compare four newly developed second generation assay systems for quantitation of CA 125 by utilizing an alternative simple approach to method comparison analysis. This alternative comprises the construction of relative difference plots and mountain plots, previously described by Krouwer et al. (Eur J Clin Chem Clin Biochem 1995; 33:525-7). In addition, the diagnostic value of the assays was illustrated through receiver-operating-characteristic (ROC) curves. Sera obtained from 300 women were assayed for CA 125 using the Abbott IMx CA 125 assay (Abbott), the Centocor CA 125 II RIA assay (Centocor), the Berilux Ov testing kit for CA 125 (Behringwerke), and the CA 125 TR-FIA assay (Wallac Oy). Both the relative difference plots and the mountain plots revealed higher serum concentrations with the Centocor RIA II (Median +33%, P2.5 to P97.5: -25% to 161%) and Berilux (Median +28%, P2.5 to P97.5: -17% to 108%) compared to the Abbott IMx system. The TR-FIA assay system showed lower serum concentrations (Median - 17%, P2.5 to P97.5: -74% to 229%). The combination of relative difference plots and mountain plots demonstrated clearly the wide range of differences between CA 125 assays measuring the same analyte. The relative difference plots provided insight into the distribution of the differences over the range of measurement as well as the identification of outliers. A simple quantitative assessment of the median differences could be made from the overlaying mountain plots. The close correspondence observed between the ROC curves illustrated that assay systems for CA 125 differing in design (type of antibodies used) and format can produce similar results on group level. However, the results of the clinical evaluation underline the importance of the application of assay specific cut-off values.

[Three patients with accidental hypothermia: customized rewarming]. / Drie patiënten met accidentele hypothermie; opwarmen op maat.

Three patients, two men aged 47 and 33 years and a woman aged 33 years, became hypothermic after an accident with body temperatures of 34.5, 26.2 and 23.1 degrees C, respectively. Two of them developed circulatory disorders, for which reanimation was performed. All three patients recovered after active external heating using among other things warmed infusion fluids, a warmed waterbed, continuous arteriovenous rewarming and rewarming with the heart-lung machine. For the treatment to be successful, the type of rewarming selected should be guided by the degree of hypothermia, the circulatory situation and the severity of the accessory injuries.

Bicycle-spoke injuries: a prospective study.

A prospective study of bicycle-spoke injuries was undertaken to assess the severity of soft tissue damage and the incidence of skeletal injuries. During a 12 month period, 59 children with spoke injuries were examined. Soft tissue damage was divided into three categories: bruising without laceration (four), bruising and superficial abrasions (49) and full thickness skin defects (six). Seventeen children had skeletal injuries: nine tibial fractures (five greenstick fractures and four spiral fractures) and eight epiphyseal injuries of the distal fibula. All fractures were treated non-operatively. The mean time for soft tissue healing was 16.5 days and the mean time for regaining full weight-bearing was 8.5 days. Soft tissue damage is the most important prognostic factor in the healing of a bicycle-spoke injury.

The use of 'ultrasensitive' prostate-specific antigen assays in the detection of biochemical recurrence after radical prostatectomy.

OBJECTIVE: To determine the gain in lead time obtained when using ultrasensitive prostate-specific antigen (PSA) assays in the diagnosis of biochemical progression after radical prostatectomy. PATIENTS AND METHODS: The post-operative PSA serum concentrations of 137 patients who had undergone radical prostatectomy were evaluated retrospectively. From these patients, 12 were selected who showed biochemical recurrence, as measured by the Hybritech Tandem-E Singlepoint PSA assay. Samples of the serum frozen at the time of the initial analysis were thawed and PSA values were remeasured by the Abbott IMx PSA assay and the Tandem-E Multipoint PSA assay. Analytical thresholds (zero-dose + 3 SD) for the Tandem-E Singlepoint, IMx and Tandem-E Multipoint assay were 1.0, 0.04 and 0.04 ng/mL, respectively. The lead time to the detection of a recurrence obtained when using the IMx and the Tandem-E Multipoint PSA assay was compared with that attained using the Tandem-E Singlepoint PSA assay. As a control, PSA values were determined in 58 serum specimens of nine patients having no evidence of recurrence after radical prostatectomy. RESULTS: All 58 control specimens had PSA levels below the analytical thresholds of the three assays, except one which had a PSA serum concentration of 0.08 ng/mL, estimated by the IMx assay. When compared with the lead time obtained with the Tandem-E Singlepoint assay, the 12 patients with a biochemical recurrence had a median gain in lead time of 327 days (range 60-627) with the IMx assay and of 369 days (range 60-639) with the Tandem-E Multipoint assay. CONCLUSION: A PSA value > 0.04 ng/mL after radical prostatectomy heralds further biochemical progression. The use of the ultrasensitive IMx and the Tandem-E Multipoint assays provided more lead time, but there is no clear evidence that this gain is necessarily of benefit to the patient.

Review on the simultaneous determination of total prostate-specific antigen and free prostate-specific antigen.

BACKGROUND: The total prostate-specific antigen (t-PSA) in serum measured by PSA assays represents the sum of free (f-PSA) and PSA complexed with alpha 1-antichymotrypsin. The f-PSA/t-PSA (F/T) ratio in prostate cancer (PCA) patients is lower than in patients suffering from benign prostatic hyperplasia (BPH). This review summarizes the current literature on the clinical relevance of measurement of the F/T PSA ratio. METHODS: Discussed are: physiology of PSA, assays for t-PSA and F/T ratio, factors which bias the F/T PSA ratio, use of F/T PSA ratio in the detection of PCA, correlation with histological features, and pathological stage. RESULTS: Using the F/T ratio in the intermediate t-PSA range, a reduction of approximately 30% in biopsies can be accomplished in the detection of prostate cancer. CONCLUSIONS: The F/T PSA ratio could become a valuable tool in the differentiation of BPH from PCA. To accomplish this goal, an international standardization not only for the t-PSA measurement but also for the F/T PSA ratio must be a priority for manufacturers of PSA assays.

Humoral anti-OV-TL 3 response after the intravenous administration of radiolabelled Fab' or F(ab')2 fragments in ovarian cancer patients.

The human anti-mouse antibody (HAMA) response was determined in the serum of patients suspected of having ovarian cancer who underwent radioimmunoscintigraphy with either 99Tcm-OV-TL 3 Fab' (n = 20) or 111In-DTPA-OV-TL 3 F(ab')2 (n = 73). Blood samples were collected prior to and at several time points post-intravenous injection. The detection of HAMA was performed with an in-house OV-TL 3 F(ab')2-based sandwich-type immunoradiometric assay (IRMA). The homologous IRMA demonstrated that 8 of 20 (40%) patients had developed HAMA responses after injection of Fab' fragments and that 14 of 73 (19%) patients had developed HAMA responses after F(ab')2 administration. The subclass of the measured HAMA was analysed in a limited number of samples, showing IgG or IgM as well as mixed responses. The kinetics of the HAMA responses varied greatly. Our study showed the relevance of the sampling time and frequency: HAMA responses can be easily underestimated with a low sampling frequency. The homologous IRMA described in this study was able to quantify the OV-TL 3-specific HAMA responses. With additional assays, the subclass of the HAMA could be further analysed. Remarkably, the fraction of HAMA responders after injection of OV-TL 3 Fab' fragments was in the same range as the proportion of HAMA responders after F(ab')2 administration.

Analytical and clinical performance of improved Abbott IMx CA 125 assay: comparison with Abbott CA 125 RIA.

We compared the improved Abbott IMx cancer antigen (CA) 125 assay (cat. no. 7A89) with the Abbott CA 125 RIA. Serum specimens were from healthy perimenopausal women (n = 124) and from patients with benign gynecologic and nongynecologic diseases (n = 124), ovarian carcinoma (n = 104), or other malignancies (n = 193). The IMx assay detected as little as 0.193 kAU/L CA 125 (AU = arbitrary Abbott unit), demonstrated up to 29% overestimation upon serum dilution, low within-assay (2.7-5.6%) and between-assay (4.8-8.2%) CVs, and no high-dose hook effect < or = 46,000 kAU/L nor influence from human anti-mouse antibodies in serum of women injected with OC 125 F(ab')2. Values by IMx were 20% lower than by RIA for healthy perimenopausal women (n = 100; IMx = 0.80 RIA - 2.5 kAU/L), and at least 50% higher for those with benign or malignant ovarian disorders at concentrations < 100 kAU/L. Receiver-operating characteristic (ROC) curve analysis of ovarian neoplasma vs perimenopausal controls indicated a gain of specificity and sensitivity with the improved IMx assay over the RIA, but ROC performance was the same with either assay if patients with benign ovarian disorders were used as controls.

Circulating inhibin levels in lactating and nonlactating women.

OBJECTIVE: To investigate the role of inhibin in the human puerperium, by measuring serum levels of immunoreactive inhibin in both lactating and nonlactating women. DESIGN: Prospective, comparative, open study. SETTING: Department of obstetrics and gynecology of a university hospital. PATIENTS: Fourteen healthy women who delivered at term: seven lactating women and seven nonlactating women treated with the dopamine-agonist CV 205-502. MAIN OUTCOME MEASURES: Serum immunoreactive inhibin, PRL, FSH, LH, E2, and P. RESULTS: All women showed a rapid decline of immunoreactive inhibin levels during the first postpartum days. Thereafter the pattern depended on the way of feeding. Nonlactating women, with their rapid return of pituitary and ovarian function, showed increasing immunoreactive inhibin levels to a maximum on day 24 (950 +/- 180 U/L). Lactating women did not show ovarian activity despite high FSH levels, and immunoreactive inhibin stayed on a low level (230 +/- 40 U/L on day 24). There was a significant correlation between immunoreactive inhibin and E2. CONCLUSIONS: The rapid decline of immunoreactive inhibin (elimination of placental hormone) is followed by an increase in nonlactating women (production by the maturing follicle) and by persistently low levels in lactating women. The lack of adequate levels of immunoreactive inhibin in lactating women may be an explanation of the relatively high FSH levels during lactation.

CA-125 levels in cervical mucus during the menstrual cycle.

OBJECTIVE: To determine CA-125 levels in cervical mucus (CM) during the menstrual cycle and their relationship to gonadal steroids and ovulation. DESIGN: Prospective study. SETTING: Two academic tertiary referral centers. PARTICIPANTS: Thirteen women with a normal fertility work-up. INTERVENTIONS: CA-125 and protein concentrations were measured in CM aspirated from the endocervical canal on alternate days in the early follicular and luteal phases and on a daily basis during the periovulatory period. Results were correlated with hormonal determinations, serum CA-125 levels, and ultrasound examination. RESULTS: Twenty ovulatory nonconceptional cycles were analyzed. Although the mean (+/- SD) concentration of CA-125 in CM (173,900 +/- 128,900 arbitrary U/mL) appeared relatively constant along the cycle, a large variation among the different samples was observed, ranging from 9,000 to 830,000 arbitrary U/mL. No clear trend could be detected as related to hormonal changes and ovulation. However, when the mucus CA-125 concentration was multiplied by the total volume of the correspondent sample, a clear periovulatory increase of total CA-125 levels was found. This was further supported by a similar trend showed by the calculated CA-125:protein concentration ratio. CONCLUSIONS: CA-125 is present in CM in high concentrations that vary widely along the cycle. Although no cyclical variation in CA-125 concentration could be determined, there was an apparent increase of total CA-125 levels parallel to the augmented mucus production during the periovulatory period. This further suggests a possible involvement of this glycoprotein in the secretory process of endocervical glands.