Home monitoring of 17 alpha-hydroxyprogesterone levels by filter paper blood spots in patients with 21-hydroxylase deficiency

BACKGROUND: 21-Hydroxylase (21-OH) deficiency is characterized by an excess of androgen in both sexes and premature skeletal maturation resulting in short adult stature and male infertility. To achieve optimal height in children and fertility in adults, the replacement treatment of 21-OH deficiency with glucocorticoids should be regulated in order to adequately reduce the excess of androgens while minimizing the dose of glucocorticoids required. Neonatal screening for 21-OH deficiency is based on the measurement of the 17 alpha-hydroxyprogesterone (17-OHP) level from blood spotted on filter paper. The aim of this study was to examine whether repeated daily blood sampling on filter paper can assist in improving the monitoring of, and compliance to, 21-OH deficiency treatment. METHODS: During a 5-year period (1989-1994) we instructed 8 patients with 21-OH deficiency (2 males with salt losing, 2 males and 1 female simple virilizing, and 1 male and 2 females with nonclassical 21-OH deficiency) aged 1.3-36 years to sample blood on filter papers 1-4 times a day and send the papers to our neonatal screening laboratory by mail. On 62 occasions we measured both serum and filter paper 17-OHP levels in order to assess the degree of correlation between the two methods. RESULTS: Comparison between the filter paper and serum 17-OHP levels showed a correlation coefficient of r = 0.87. The filter paper levels were almost always higher than the serum levels. The serum 17-OHP levels were < 1 ng/ml whenever the filter paper levels were < 3 ng/ml. On long-term follow-up of 17-OHP filter paper levels we observed major diurnal and day-to-day fluctuations which might not have been noticed on routine follow-up clinic visits. CONCLUSIONS: Filter paper follow-up of 17-OHP levels can assist in optimizing the replacement treatment in patients with 21-OH deficiency while reinforcing compliance and decreasing the need for frequent clinic visits and hospitalizations. By adjusting the glucocorticoid type, dose, and time of administration to each patient we should be able to achieve optimal growth without bone age acceleration in children, to avoid overtreatment, and to improve fertility in adults.     

Determination of glucose levels using dried filter paper blood spots: new perspective in home monitoring

We present a method for the determination of blood glucose using dried filter paper blood spots. To validate this method, we compared our results using filter paper and simultaneously collected venous blood. We demonstrated that there is a linear relationship between the filter paper glucose levels and those determined in whole blood (r = 0.98). There was no significant difference between the results of the two methods (p > 0.05). This method is a cheap alternative which may improve the control of diabetes mellitus, and may also be very useful in the diagnosis of postprandial hypoglycemia and other special situations.     

Rapid method for screening dried blood samples on filter paper for human immunodeficiency virus type 1 DNA

PCR is a highly sensitive method for the detection of human immunodeficiency virus type 1 (HIV-1) nucleic acids in blood mononuclear cells and plasma. However, blood separation techniques require extensive laboratory support systems and are difficult when a limited volume of blood is available, which is often the case for infants. The use of blood samples stored on filter paper has many advantages for the detection of perinatal HIV-1 infection, but current methods require extraction and purification of target DNA prior to PCR amplification. We report a highly sensitive and rapid method for the extraction and detection of HIV-1 DNA in infant blood samples stored on filter papers. Because this rapid protocol does not involve steps for the removal of potential inhibitors of the PCR, the highest sensitivity is achieved by testing the filter paper lysate in quadruplicate. Assays for HIV-1 DNA were done by using nested PCR techniques that amplify HIV-1 gag DNA from blood spot samples on filter paper and from corresponding viably frozen mononuclear cells separated from venous blood samples obtained from 111 infants born to HIV-1-seropositive mothers. PCR results with blood from filter papers showed 100% specificity (95% confidence internal [CI] 93.1 to 100%) and 96% (95% CI, 88.65 to 98.9%) and 88% (95% CI, 79.2 to 94.5%) sensitivity (for quadruplicate and duplicate tests, respectively) compared to PCR results with blood mononuclear cells. Moreover, this method could detect HIV-1 sequences of multiple subtypes.     

Estimation of galactose-1-phosphate in blood spotted on filter paper

A method is described for the enzymic estimation of galactose-1-phosphate (Gal-1-P) in blood which has been applied to filter paper and allowed to dry. The successful clinical management of patients with galactosemia depends upon exclusion of galactose from their diet. Earlier studies demonstrated that red cell Gal-1-P is a sensitive indicator of exposure of such patients to galactose. These earlier methods, however, required venipuncture, preparation of washed, packed red cells, and shipment of the sample in dry ice to a central laboratory. With the present method, capillary blood can be drawn by a nonphysician, applied to filter paper and mailed in a conventional envelope at ambient temperature. From this sample, the Gal-1-P content of the red cells can be determined, if the hematocrit is known. These conveniences should allow estimates of Gal-1-P at a frequency more appropirate for optimal dietary control.     

The indirect hemagglutination test for onchocerciasis performed with blood collected on filter paper

In Guatemala, an indirect hemagglutination (IHA) test for onchocerciasis, performed with a crude saline extract of Onchocerca volvulus and blood samples taken on filter paper, showed a high level of sensitivity and specificity. IHA titers of blood samples from the ear lobe taken on filter paper and of sera obtained by venipuncture showed a high correlation.     

A routine method for thin layer chromatographic determination of urocanic acid in blood samples impregnated on filter paper cards (author's transl)

A method is described for mass screening at birth for histidinaemia. Blood samples are collected on filter paper cards, as used in the Guthrie test. These samples are transferred, together with standards of histidine and urocanic acid, to thin layer chromatography plates by a type of sandwich technique. The plates can be evaluated semiquantitatively, after development and detection with Pauly's reagent. The detection limit for both substances is less than 10 mg/1 (60 mumol/1). Greater sensitivity and absence of interference, greatly reduce the proportion of equivocal results compared with the Guthrie test.     

Detection of IgM against the dengue++ virus in whole blood absorbed on filter paper]

Thirty-four sheep were submitted to surgery substituting the native ACL with the central third of the patellar tendon, ten enter this study. The purpose was to find a possible relationship between tissue pO2 and healing processes considering also the biomechanical and histomorphological aspects of the grafts. Four of them were sacrificed under general anaesthesia after 6 months, and six after 1 year in order to perform tissue pO2 measurement and an analysis of microvessel density on specimens of the normal ACL and the graft. Our data showed higher pO2 values of the autografts after 6 months. After 1 year the data was comparable to those of native ACL. This was confirmed by a microvessel count of the histological specimens and the data was in relationship to biomechanical and histomorphological analysis. Tissue pO2 can be observed and recorded in "in vivo" ACL, and patellar tendon used as graft, with no injury to their integrity. The monitoring system might be considered as an experimental tool for indirect controls of the anterior cruciate substitutes.     

The use of whole blood absorbed on filter paper to detect Wuchereria bancrofti circulating antigen

The Og4C3 enzyme-linked immunosorbent assay (ELISA) to detect circulating Wuchereria bancrofti antigen uses 50 microL of serum. In this study, a whole blood sample absorbed on filter paper was tested as a substitute for serum. Serum samples were obtained from 60 Sri Lankan subjects by venepuncture and finger-prick blood samples from the same individuals were directly absorbed on filter paper. Og4C3 ELISAs using serum and filter paper blood were compared. Despite the fact that the estimated amount of serum available for the ELISA with filter paper blood was only one-fifth of that available when serum was used, the 2 ELISAs gave almost identical results. Of the 39 positive serum samples, 38 were detected using filter paper blood. Employing the ELISA using filter paper blood, 619 people in Matara, Sri Lanka, were examined for antigenaemia. The positivity rate was 22.5%, 3.1 times higher than the rate of microfilaraemia detected by examination of 60 microL blood films.     

Examination of 17 beta-estradiol and progesterone levels in peritoneal fluid and blood serum of women with endometriosis

Authors have presented and assessment of estradiol and progesterone levels in peritoneal fluid and blood serum in women with endometriosis. Peritoneal fluid was collected during laparoscopy performed in luteal phase of the cycle. In this cycle ovulation was controlled in all women. An ovulation was confirmed ultrasonographically and laparoscopically in 45% of women with endometriosis and in 80% of that without the illness. Progesterone concentration in peritoneal fluid in women with endometriosis was significantly lower to the control (p < 0.01).     

Retrospective diagnostics of congenital cytomegalovirus infection performed by polymerase chain reaction in blood stored on filter paper

OBJECTIVE: External reference points, particularly Kirschner pins (K-wire), placed in the region of the nasion have been shown to improve the accuracy of maxillary vertical repositioning. Although no complications associated with this technique have been reported, there is a potential for injury to the anterior cranial fossa or frontal sinus. The purpose of this study was to measure the shortest distance from the nasion to the anterior cranial fossa and from the nasion to the frontal sinus. These measurements were used to establish anatomic guidelines governing safe placement of external reference point pins. STUDY DESIGN: Twenty-seven cadaver heads were sectioned in the midsagittal plane for gross study. Using a Boley gauge, two specific measures were obtained: (1) distance from deepest depression of nasion to the most anterior and inferior projection of the anterior cranial fossa, and (2) distance from nasion to the most inferior aspect of the frontal sinus. All measurements were made in the midsagittal plane. RESULTS: The average distance from nasion to anterior cranial fossa was 16.9 mm (range 13.0 to 20.0 mm) and the smallest distance, 13.0 mm, was seen in two specimens. The average distance from nasion to the frontal sinus was 6.2 mm (range 2.0 to 10.0 mm) and the smallest distance, 2.0 mm, was seen in three specimens. CONCLUSION: Based on our findings, we recommend the following: (1) place pin to a depth of no more than 8 mm into bone, (2) place pin 5 to 10 mm inferior to soft tissue nasion, and (3) place pin in an anterosuperior to posteroinferior direction (i.e., roughly perpendicular to the nasal dorsum). When these anatomic guidelines are followed, one would expect minimal morbidity associated with the placement of ERP pins.     

Adaptation and validation of antibody-ELISA using dried blood spots on filter paper for epidemiological surveys of tsetse-transmitted trypanosomosis in cattle

The indirect enzyme-linked immunosorbent assay (ELISA) for the detection of anti-trypanosomal antibodies in bovine serum was adapted for use with dried blood spots on filter paper. Absorbance (450 nm) results for samples were expressed as percent positivity, i.e. percentage of the median absorbance result of four replicates of the strong positive control serum. The antibody-ELISA was evaluated in Zambia for use in epidemiological surveys of the prevalence of tsetse-transmitted bovine trypanosomosis. Known negative samples (sera, n = 209; blood spots, n = 466) were obtained from cattle from closed herds in tsetse-free areas close to Lusaka. Known positive samples (sera, n = 367; blood spots, n = 278) were obtained from cattle in Zambia's Central, Lusaka and Eastern Provinces, diagnosed as being infected with Trypanosoma brucei, T. congolense, or T. vivax using the phase-contrast buffy-coat technique or Giemsa-stained thick and thin blood smears. For sera (at a cut-off value of 23.0% positivity) sensitivity and specificity were 86.1 and 95.2%, respectively. For bloodspots (at a cut-off value of 18.8% positivity) sensitivity and specificity were 96.8 and 95.7%, respectively. The implications of persistence of antibodies following treatment or self-cure are discussed.     

Antithrombin, protein C and protein S levels in 127 consecutive young adults with ischemic stroke

Children with spina bifida, cerebral palsy, mental retardation, developmental delays, and seizure states are handicapped with sensorimotor deficits, including gait or coordination instability, temperature insensitivity, and mental simplicity. These handicaps make this distinct and unpretentious population more susceptible to lethal burns. A 30-year review was conducted in a pediatric burn center to examine the relationships between pediatric sensorimotor deficit and burn injury. Of the 4874 acute burn admissions, 66 children were identified with preexisting sensorimotor deficits. Data indicate that children with sensorimotor deficit are more prone to burn injury from both their physical impairment and poorly supervised environments. In addition to extended hospitalizations, these children bear significantly higher mortality risks. Had the special supervisions and protection required by such handicapped children been provided, 80% of the burn injuries could have been prevented. Results emphasize that the future of these special children with sensorimotor deficits relies on health care providers playing a greater role in educating parents and caregivers.     

剪切式切纸技术在PASSIM系列卷接机组上的应用研究

研究了国内外主要卷接机组水松纸切割系统,剪切式切纸较滚切式切纸具有明显的技术优势.通过分析PROTOS系列切纸轮的工作原理,设计了一种机构应用于PASSIM系列卷接机组中,既保留了PASSIM系列切纸轮在一个轮体上实现切纸与搓烟2种功能的特点,同时在PASSIM系列卷接机组中实现了剪切式技术.     

Biliary excretion in persons with low blood glutathione levels

To examine their possible predictive value for the development of asthma, the serum concentration of eosinophil cationic protein (ECP) and the total eosinophil count were measured at admission in 25 children aged 1-17 months hospitalized for their first episode of bronchiolitis. After an average of three years the parents of 23 index patients answered a questionnaire to determine development of asthma. Eight children were defined as having asthma at follow-up based on at least three episodes of wheezing. The remaining 15 children had experienced only one or two episodes of wheezing, and all of these children had been wheeze free for the last year. The serum concentrations of ECP were similar in children who subsequently developed asthma (8.0 microg/l; 3.6 to 14.2 (median; quartiles)) and in those who did not (12 microg/l; 4.5 to 16.8). Moreover, the total eosinophil counts were similar in asthmatic (0.10 x 10(9)/l; 0.04 to 0.20) and non-asthmatic patients (0.09 x 10(9)/l; 0.02 to 0.13). In conclusion, our study suggest that neither the serum concentration of ECP nor the total eosinophil count can predict the development of asthma when measured in children admitted for their first episode of bronchiolitis, but larger studies need to be carried out to confirm these results.     

Does collecting repeated blood samples from each subject improve the precision of estimated steroid hormone levels?

Peripheral blood mononuclear cells were isolated from noninfected control cows and from cows with either subclinical or clinical paratuberculosis (Johne's disease). Cells were incubated for 6, 12, 24, and 48 hours in complete medium with the following mitogens: concanavalin A (ConA), phytohemagglutinin-P (PHAP), pokeweed mitogen (PWM), and Escherichia coli lipopolysaccharide. In addition, cells were incubated for the same time periods with a Mycobacterium paratuberculosis sonicate (MpS) and live and heat-killed M. paratu-berculosis at 10:1 bacteria: cell ratio. After incubation, cell-free supernatants were analyzed for gamma-interferon (gamma-IFN) production. Cells from subclinical cows produced significantly higher levels of gamma-IFN than did cells from clinical animals after stimulation with mitogens ConA, PHAP, and PWM. Levels of gamma-IFN produced by noninfected control animals generally followed the pattern of those of subclinical animals. After incubation with MpS, significantly greater quantities of gamma-IFN were produced by cells isolated from subclinical animals than by cells from clinical cows and noninfected controls. Stimulation of cells with heat-killed or live M. paratuberculosis evoked a similar response. This study indicates that gamma-IFN production by peripheral blood mononuclear cells in response to M. paratuberculosis antigen may be an important diagnostic tool for the detection of paratuberculosis in subclinically affected animals.     

High-performance liquid chromatographic method for the simultaneous determination of mefloquine and its carboxylic metabolite in 100-microliters capillary blood samples dried on paper

A reversed-phase high-performance liquid chromatographic method is described for the analysis of mefloquine and its carboxylic metabolite in 100-microliters capillary blood spots dried on chromatographic paper. Each spot was cut into small pieces, and mefloquine and its metabolite were eluted with an ammonia-water solution (10:90, v/v). The compounds were extracted simultaneously after alkalization at pH 9.5 using tetrabutylammonium as ion-pairing agent and then separated on a C18 column with ultraviolet detection at 227 nm. The recovery of the drugs from spiked blood applied to paper and dried was 70-80%, and the inter-assay precision at 1.0-5.0 mumol/l (therapeutic range) was less than 10%. The correlation between extractions from venous whole blood and capillary blood applied to chromatographic paper was more than 0.94. The analytes were stable in dried blood spots for at least fifty days at -20 degrees C. The decrease of concentration was less than 10%, when the paper was stored at 37 degrees C for fifty days. The assay is reliable and easy to use for therapeutic monitoring of mefloquine with a lower limit of determination of 0.3-0.5 mumol/l.     

X射线荧光光谱滤片法测定镀铑液中铑含量

采用可调移液枪移取样品溶液,制成溶液滤片,以波长色散X射线荧光光谱仪直接测定了铑镀液中铑的含量。实验表明,利用溶液滤片法绘制的校准曲线线性相关系数为0.999 1,与传统方法制得的校准曲线线性相关系数相一致,且不同体积的溶液在纸片上的扩散不会对测量结果造成影响。该方法样品用量少,消除了基体效应,且校准曲线能够保证长期使用。方法的检出限为11.52 μg。对实际镀铑液样品进行测试,结果同电感耦合等离子体质谱法和火焰原子吸收光谱法测量结果一致,相对标准偏差＜2.1%。     

Estimation of numbers of malaria clones in blood samples

Methods are derived for estimating the mean number of clones of the haploid malaria parasite Plasmodium falciparum from samples of blood of infected hosts which have been tested for the presence of alleles at marker loci. For example, at a locus with three alleles the sample might contain only A1, or A1 and A2, or A1, A2 and A3, with multiple allele classes being more common at high infection rates. Assuming either a Poisson or negative binomial distribution of numbers of infections per host, formulae are derived for the frequency of different classes of blood samples, and maximum likelihood methods are used to estimate the mean number of clones and allele frequencies. Two data sets, each on two loci, are analysed. One data set was from the same locality in Tanzania from which oocysts of the parasite in mosquito vectors were tested for clonality (i.e. diploid unions of gametes from the same clone) using genetic markers. Good agreement was obtained between the observed clonality in oocysts and that expected from the number of infections per host (mean approximately three).