Orienting responses to various visual stimuli in children with visual processing impairments or infantile nystagmus syndrome.

Quantification of orienting responses can be used to differentiate between children with cerebral visual impairment and infantile nystagmus syndrome. To further improve the sensitivity of this method, we compared orienting responses to a Cartoon stimulus, which contains all sorts of visual information, to stimuli that contain only Contrast, Form coherence, Motion coherence, Color and Motion detection. The stimuli were shown on an eye tracker monitor using a preferential looking paradigm. We found that both groups of children showed general slowing in orienting responses compared to controls. The children with cerebral visual impairment had significantly prolonged responses to Cartoon compared to the children with nystagmus, whereas the children with nystagmus had prolonged responses to Motion detection and larger fixation areas. Previously reported differences in orienting responses to Cartoon were replicated. Application of specific visual information did not alter the sensitivity of the method to distinguish between children with visual processing deficits.

Inserting and manipulating DNA in a nanopore with optical tweezers.

The translocation of small molecules and polymers is an integral process for the functioning of living cells. Many of the basic physical, chemical, and biological interactions have not yet been studied because they are not directly experimentally accessible. We have shown that a combination of optical tweezers, single solid-state nanopores, and electrophysiological ionic current detection enable deeper insight into the behavior of polymers in confinement. Here we describe the experimental procedures that are necessary to manipulate single biopolymers in a single nanopore, not only by electrical fields, but also through mechanical forces using optical tweezers.

Evaluation of automated nucleic acid extraction devices for application in HCV NAT.

BACKGROUND: To further improve the safety of the blood supply, various national blood transfusion organizations presently use or are in the process of implementing routine HCV NAT in minipools. According to the Committee for Proprietary Medicinal Products (CPMP) of the European Union, the HCV NAT detection limit of the assay should be 100 IU per mL (270 geq/mL) for testing initial plasma pools. Paul Ehrlich Institute (PEI) regulations stipulate that 5000 IU per mL (13,500 geq/mL) must be detected to calculate the amount contributed by individual donations composing the minipool. The sensitivity for HCV RNA extraction achieved by three commercially available laboratory kits was compared. STUDY DESIGN AND METHODS: Nucleic acids from 1-in-3 serial dilutions of an HCV RNA run control (Pelispy, CLB) were extracted with three kits (Cobas Amplicor, Roche Diagnostic Systems; BioRobot 9604, Qiagen; and NucliSens Extractor, Organon Teknika). HCV PCR of all extracts was performed using a second-generation Cobas Amplicor HCV test and the Cobas Amplicor analyzer. RESULTS: The manual Cobas Amplicor, the BioRobot 9604, and the NucliSens Extractor setups allow a 95-percent HCV RNA detection limit of 129, 82, and 12 geq per mL, respectively. The maximal pool size for the manual Cobas Amplicor, the BioRobot 9604, and the NucliSens Extractor kits that would still meet the PEI criteria for HCV NAT in minipools was calculated at 104, 164, and 1125 donations, respectively. CONCLUSION: All three HCV NAT kits evaluated meet the criteria set by CPMP and PEI. The highest sensitivity for HCV NAT screening can be achieved with the high-volume NucliSens Extractor method in combination with the Cobas Amplicor HCV v2.0 test on the Cobas Amplicor analyzer.

Development of an on-line coupling of liquid-liquid extraction, normal-phase liquid chromatography and high-resolution gas chromatography producing an analytical marker for the prediction of mutagenicity and carcinogenicity of bitumen and bitumen fumes.

A fast and fully automated system for the determination of polycyclic aromatic compounds (PACs) is described. The system has been developed to produce an analytical 'marker', correlating chemical characteristics (including PAC analysis) with mutagenicity and carcinogenicity. The products of interest are bitumen fumes, bitumen and other (heavy or even residual) oil products, regardless of their boiling range. Dimethyl sulphoxide (DMSO) extractables obtained from a flow-injection analysis (FIA) system are introduced on-line in a normal-phase liquid chromatographic (NPLC) system. Here, the PACs are separated from the DMSO and possible co-extracted heavy residual species. The final step incorporates on-line gas chromatographic analysis of the three-to-six-ring PAC fraction, followed by flame-ionisation detection for quantification. It was demonstrated that data obtained from samples in the distillate lubrication-oil range correlate well with data obtained from the manual DMSO-extraction method standardised by the Institute of Petroleum as IP346.

Cognitive-behavioral therapy for noncardiac chest pain: a randomized trial.

PURPOSE: Patients with nonischemic chest pain frequently experience recurrent symptoms, have persistent functional and occupational disability, and are high utilizers of health-care resources. Our aim was to evaluate the efficacy of a cognitive-behavioral treatment for patients with noncardiac chest pain. PATIENTS AND METHODS: Subjects were recruited from patients with at least weekly episodes of noncardiac chest pain, as diagnosed by a cardiologist. The main outcome measures were frequency and intensity of chest pain at 6 and 12 months. RESULTS: Seventy-two patients were enrolled in the study; 37 were assigned to cognitive-behavior therapy and 35 to usual care. Sixty-five patients completed the study. Intervention patients improved significantly with regard to frequency and intensity of chest pain: 15 (48%) of the 31 patients in the treatment group were pain free at 12-month follow-up compared with 4 (13%) of the 33 patients in the control group (P = 0.002). CONCLUSIONS: Cognitive-behavioral therapy for noncardiac chest pain patients was effective compared with usual care.

Family treatment, expressed emotion and relapse in recent onset schizophrenia.

A controlled longitudinal treatment study was carried out to investigate the effect of a behavioral family treatment on Expressed Emotion (EE) and to examine the correspondence between EE changes and relapse rates. Subjects were 52 patients with recent onset schizophrenia or related disorders and their parents. After completion of inpatient treatment they were randomly allocated to individual treatment or individual treatment plus family treatment. The family treatment consisted of education and training in communication and problem-solving skills. Expressed Emotion was measured with the Five-Minute Speech Sample (FMSS). The findings show that family treatment did not have a significant positive effect on EE level. The dichotomous FMSS/EE did not systematically change and these findings were comparable with the results of prior EE research. A scoring system that included all subscores of the FMSS was somewhat more sensitive to changes. In the individual treatment condition relapse rates tended to co-occur with a change in FMSS/EE level, irrespective of the direction of this change.

Treatment, expressed emotion and relapse in recent onset schizophrenic disorders.

The effect of in-patient and individual orientated psychosocial intervention (IPI) and in-patient and individual and family orientated intervention (IPFI) across levels of expressed emotion (EE) on relapse was compared in a group of patients with recent onset schizophrenic disorders. Patients were randomly assigned to an individual orientated psychosocial intervention programme or to an identical psychosocial programme plus a behavioural family intervention. Seventy-six patients were studied during a 12 month out-patient treatment period after an in-patient treatment programme in which parents followed a psychoeducational programme. Overall relapse rates during the out-patient interventions were low (16%). Adding family intervention to the psychosocial intervention did not affect the relapse rate. Patients in low EE families relapsed slightly more often during the psychosocial plus family intervention. In-patient treatment with psychoeducation for parents, followed by an out-patient psychosocial intervention programme, has a favourable impact on relapse. Additional family intervention may increase stress in low EE families, thus affecting relapse in their children.

Effects of chronic factor VIII substitution on immune parameters in HIV seronegative haemophiliacs: a comparison between cryoprecipitate and factor VIII concentrate.

Chronic substitution therapy of HIV-negative haemophiliacs with factor VIII products can result in abnormalities of ex-vivo measured immune parameters. To assess a possible relation between these abnormalities and product purity, we analyzed two groups of HIV-negative HCV-positive haemophiliacs, one treated with cryoprecipitate exclusively, the other with more purified factor VIII concentrates. Compared to age matched non-transfused male controls, increased numbers of white cells, granulocytes, IgG and IgM levels and decreased CD4+/CD8+ ratios were found in both patient groups. In the concentrate receivers, the numbers of mononuclear cells, CD4+, CD8+ and CD3+/HLA-DR+ cells indicating activated T-cells, were higher than in the cryoprecipitate group. In conclusion, both cryoprecipitate and intermediate/high purity concentrate recipients showed immune parameter abnormalities. These abnormalities tended to be somewhat more pronounced in patients treated with concentrates. By now there is no indication of the clinical relevance of the abnormalities in previously treated HIV seronegative haemophiliacs.

Ovarian activity suppression by two different low-dose triphasic oral contraceptives.

In an open, randomized study in an outpatient clinic of a large teaching hospital, thirty-one female volunteers with regular cycles and established ovulation by ultrasonography were given one of two triphasic oral contraceptives containing ethinylestradiol combined with levonorgestrel or desogestrel during six cycles of treatment. The main outcome measures were transvaginal ultrasonography and serum E2 and P measurements in pill cycles 1, 3 and 6. No ovarian activity was found in 10 subjects. Among the remaining 21 women who showed ovarian activity, most follicle-like structures developed in the pill-free week and decreased in size or disappeared in the first pill week. One women taking triphasic desogestrel had evidence of a luteinized unruptured follicle and one women taking triphasic levonorgestrel had a possible ovulation. The latter women also showed symptoms of lower abdominal pain. A statistically significant difference in ovarian activity between the two oral contraceptives could not be established. The two triphasic oral contraceptives suppressed ovarian activity to the same degree. A trend was seen towards increasing ovarian activity with duration of use in both treatment groups.

PIP: Oral contraceptives (OC) have an inhibitory effect upon ovarian activity and the quality of cervical mucus and endometrium. Low-dose OCs, however, do not completely suppress ovarian function. This study was conducted to compare ovarian activity over a 6-month period during treatment with two low-dose triphasic OCs containing ethinylestradiol combined with levonorgestrel or desogestrel. 31 female volunteers aged 20-35 years with regular cycles and established ovulation were given one of the OCs over 6 cycles of treatment. Main outcome measures were transvaginal ultrasonography and serum E(2) and P measurements in pill cycles 1, 3, and 6. No ovarian activity was found in 10 subjects. Among the remaining women who showed ovarian activity, most follicle-like structures, indicative of ovarian activity, developed during the pill-free week and decreased in size or disappeared during the first pill week. One woman taking triphasic desogestrel had evidence of a luteinized unruptured follicle, while one woman taking triphasic levonorgestrel had a possible ovulation. This latter woman also showed signs of lower abdominal back pain. No statistically significant difference in ovarian activity could be established between the two OCs, with the two contraceptives suppressing ovarian activity to the same degree. A trend was seen toward increasing ovarian activity with duration of use in both treatment groups.

Surviving gynecologic cancer and coping with sexual morbidity: an international problem.

A review of the international literature reveals that significant sexual morbidity occurs for gynecologic cancer survivors, and that there has been little reduction in either the incidence or the severity of problems over the last 50 years available to research. In fact, prospective data from the last 10 years highlight the discrepancy in sexual activity and sexual responsiveness and the higher frequency of sexual dysfunctions for patients who have been treated for gyneclogic cancer in comparison to age-matched healthy counterparts. We conclude with an international summary and discuss specific research directions for disease sites. We urge international efforts to address quality of life concerns of gynecologic cancer survivors.

Preparation of lyophilized heat-treated cryoprecipitates from a small pool of plasma obtained by apheresis.

A method was developed for the batch-wise production of small-pool lyophilized heat-treated cryoprecipitates in a regional Blood Bank. Ten vials of final product were derived from twenty 600 g apheresis plasma donations. We found 384 IU of factor VIII per vial and a specific activity of 0.20 IU per mg total protein. Production recovery was 340 IU of factor VIII per kg plasma. The method was easy to perform and gave a product of good quality.

Quality control programme for the preparation of small-pool lyophilized heat-treated cryoprecipitates.

International and national documents on and standards for quality control have been introduced for Blood Banks. Recently, the Standard Registration Document and a National Health Authority Licence for factor VIII preparations based on the Document were introduced in the Netherlands. In the course of developing a preparation of lyophilized heat-treated cryoprecipitates in our Blood Bank, we used this Standard Registration Document and the good pharmaceutical manufacturing practices to design a quality control protocol. The aim of this protocol was to provide documented evidence on the quality of both our product and the production process. The protocol included the validation or revalidation of individual installations and procedures used during production, validation of the whole process and quality control of routine production. With our quality protocol we have been able to demonstrate that our preparation is consistently of the intended quality.

Evaluation of six enzyme immunoassays for antibody against human immunodeficiency virus.

Six commercial enzyme immunoassays (EIA) were evaluated in 6488 serum samples, with immunoblot analysis as the confirmatory test for antibodies against human immunodeficiency virus (HIV). The Abbott and Wellcome tests identified all 163 immunoblot-positive samples correctly, whereas the other tests did not detect 1-3 samples. In AIDS patients (predominantly with antibodies to gp41env) Organon's EIA was less sensitive (p less than 0.05) and Wellcome's more sensitive (p less than 0.05) than the immunoblot assay. In symptom-free anti-HIV-positive subjects (antibodies to almost all viral antigens and high titres of anti-p24gag) all the EIA were significantly (p less than 0.05) less sensitive than the immunoblot assay. The frequencies of false-positive reactions in a "tricky" panel of samples from patients with autoimmune and acute viral diseases and in a blood-donor panel were Abbott 9.5%, 0.42%: Organon 1.7%, 0%; Litton 1.0%, 0.4%; Behring 2.7%, 0.06%; Wellcome 0%, 0%; and Pasteur 0%, 0.02%. The results of a seventh EIA (Dupont) were excluded from the study at the company's request. All six EIA evaluated are suitable tests for anti-HIV screening in samples from patients and blood donors.

Starting a plasmapheresis program with hollow-fiber filtration.

In a routine donor center, a program of hollow-fiber filtration can be instituted. With the Organon Teknika PLASMAPUR system the collection of 600 ml plasma was completed within 50 min in 76% of the procedures. Changing the tubing set took 12-15 min. From the plasma a frozen cryoprecipitate is made with a recovery of 59%. After pooling, lyophilization and heat treatment, the recovery is 42%.

Influence of the primary anticoagulant on the recovery of factor VIII in cryoprecipitate.

The influence of anticoagulant on overall factor VIII-yield was measured by drawing blood from one donor simultaneously in three bags containing ACD, CPD and heparin, respectively. After parallel processing factor VIII:C and factor VIII:CAg were measured. It is concluded that, under the circumstances used in this experiment, CPD gives the highest yield of factor VIII.