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Introduction: Alternative methods of dry storage and transportation may be a viable alternative to the use of liquid storage medium for cervical samples, especially for screening programs in places with few resources., Objective: The objective of this study is to verify the viability and efficacy of human papillomavirus DNA (HPV-DNA) detection in cervical cell samples collected and stored on a Flinders Technology Associates (FTA) card (Whatman Indicating FTA® Elute Micro Card) and subsequently recovered in ethanol-based liquid medium and to compare the results to those obtained using samples stored directly in ethanol-based liquid medium., Study Design: Thirty-four women submitted to ETZ (excision of the transformation zone of the cervix) were included in this study. Before ETZ, 2 samples of exfoliated cervical cells were collected from each woman by a doctor and stored in ethanol-based liquid medium and on an FTA card. DNA recovery from FTA samples was performed using ethanol-based liquid medium. Detection of HPV-DNA in the samples was performed using the Cobas® 4800 HPV Test Platform., Results and Conclusions: The HPV-DNA detection positivity rates were 70.6% for the samples collected directly in liquid medium and 64.7% for the samples stored on the FTA card, with high detection accuracy in the DNA samples recovered from the FTA card (area under the curve = 0.958; 95% confidence interval = 0.890-1.000). The concordance between the results obtained using the 2 storage media was 94.1% (Kappa = 0.866). These preliminary results suggest that collection of cervical material on an FTA card may be an alternative to storage in liquid medium since the liquid medium has some limitations. In addition, DNA recovery from the card using ethanol-based liquid medium streamlines the workflow in the laboratory and reduces the cost associated with reagents, thereby facilitating access to the HPV test in places with few resources and potentially improving cervical cancer screening., (© 2021 S. Karger AG, Basel.)

Introduction: The probability of prostate cancer detection is related to the amount of tissue represented. For optimal tissue representation, the specimens should preserve their regular cylindrical shape and avoid artefacts and deformation caused by fixation and preembedding. The aim of our study was to compare contemporary preembedding methods including a new method of using thick cardboard., Materials and Methods: To compare the preembedding methods, we took 36 nonfragmented cores from fresh prostatectomy specimens for each method, fixed them in formalin and made histological slides. The comparison criteria were a core section area in the middle section and the number of fragments per core after processing., Results: Two methods (preembedding on the edge of thick cardboard and on biplicated paper) provided a bigger section area of specimens. The differences in amounts of fragments were very small among the methods mentioned above and the preembedding glass with grooves, but preembedding between two sponges in a histological cassette showed higher fragmentation., Conclusions: Preembedding on the edge of thick cardboard and on biplicated paper can be recommended as effective methods of prostate biopsy core fixation. Paper or cardboard for fixation of prostate biopsy cores should be presoaked with formalin or normal saline., (© 2015 S. Karger AG, Basel.)

Background: The rapid and reliable identification of biomarkers in the smallest possible amount of blood remains a challenge in biomarker epidemiological research involving preterm newborns., Objective: We wanted to explore whether the proteomics approach of 'surface-enhanced laser desorption/ionization-time of flight mass spectrometry' (SELDI-TOF MS) is possible and feasible in whole cord blood previously dried on filter paper., Methods: Umbilical cord blood from 7 healthy newborns was frozen as serum, whole blood (with or without additives), or dried on filter paper (with or without additives). We used the SELDI-TOF MS technique for protein detection on the ProteinChip arrays: weak cationic exchange array (CM10), hydrophobic array (H50), and strong anion exchange array (Q10). Profiles were compared in terms of peak intensity and number of resolved peaks., Results: Dried neonatal blood, eluted from filter paper, revealed profiles similar to the profiles derived from serum at a protein range of 3-10 kDa. Among additives, heparin led to highest peak intensities for both blood and dried blood. Spectra from heparinized whole blood and heparinized dried blood from the umbilical cord of 8 different healthy newborns on three different types of ProteinChip arrays were very similar., Conclusion: We conclude that it is possible and feasible to use SELDI-TOF MS for recovery and detection of whole proteins from dried blood collected on filter paper. The method is easy to perform in large groups of newborns, minimizing the amount of blood needed for biomarker studies. The validity and reproducibility of this method needs to be studied in detail.

The measurement of insulin-like growth factor I (IGF-I) from extracted whole dried blood spots on filter paper is a simple method allowing sample collection to be carried out by the patient at home. One drop of blood is applied to a filter card, air dried and posted to the laboratory. The IGF-I is dissociated from its binding protein by an acidification and neutralization procedure, and IGF-I is then measured by an enzyme-linked immunosorbent assay. Results from the analysis of IGF-I using blood spots on filter paper compared favourably with those from conventional methods, thus showing it to be an ideal and cost-effective technique for the measurement of IGF-I in both adults and children., (Copyright 2001 S. Karger AG, Basel)

A number of different designs of cells have been developed for the study of dissolution processes in packed apatite powders. Basic design requirements were that no deleterious processes, such as high temperature sintering or binding agents, were involved, and that there was maximum opportunity for experimental study. One design used alternating filter paper discs and apatite layers (typically 42 mg). At the end of an experiment, the cell could be disassembled and infrared spectra and X-ray powder diffraction patterns made of individual layers. Cells without filter paper discs were also made, but terminal sampling at well-defined depths was more difficult or impossible. The cells were constructed from poly(methylmethacrylate) so the course of dissolution could be monitored by radiography. Subsurface loss of apatite was almost always seen after from 2 weeks to 8 months exposure to buffer solutions at pH 3.0-5.5. The greatest loss of apatite was typically 0.4-2 mm below the surface, which is at a larger distance than usually seen for dental enamel. This may be attributable to the low packing density (typically 35-60 vol%) found in the present systems compared with enamel.

Objective: To determine the prevalence of fiber contaminants, to provide morphologic descriptions of various fiber types and to try to understand the significance of their presence in cervical smears., Study Design: Fibers from cotton swab tips, hair, cardboard slide transporters, Cytobrush bristles and three commercial brands of tampons were smeared onto glass slides, Papanicolaou stained and examined. Then a prospective study of consecutively screened cervicovaginal smears from 1,368 women was undertaken to identify fiber contaminants., Results: Fibers were identified in 178 cases (13%). In 120, the fibers were considered extrinsic, commonly having the microscopic features of cardboard. In 58, the fibers were considered to be intrinsic to the smear because of their presence in the same plane and alignment as the cells, an associated inflammatory cell reaction, and adherent or absorbed cellular products, occasionally hemosiderin. The most common intrinsic fibers were cotton and rayon. No statistically significant relationship was identified for the presence of intrinsic fibers and patients' ages, stated menstrual dates or pathologic lesions., Conclusion: Fiber contaminants are commonly present in cervicovaginal smears, usually do not originate in the patient (are "extrinsic") and have no pathologic significance. Knowledge of the microscopic features of fiber contaminants enables them to be identified confidently and distinguished from fungal hyphae and other filamentous pathogens.

Background and Design: A controlled open trial was performed in 40 individuals suffering from chronic perianal eczema. Various toilet paper preparations representing the conventional type made of pure cellulose, the recycled and the moist type were investigated using the patch test as were allergens possibly related to toilet paper. In addition, a chronic use test and a repeated rubbing test were performed with a conventional and a recycled toilet paper preparation., Results: A total of 20 patients could be evaluated in full. A variety of reactions were seen in the patch test, most of which were of the allergic type. Allergens included many different chemical entities, e.g. preservatives used in moist toilet paper such as Kathon CG and Euxyl K 400. In a volunteer allergic to Euxyl K 400, allergy to the moist toilet paper regularly used by him was established. After discontinuation of its application, perianal dermatitis disappeared. In the repeated rubbing test at the forearm, 10 volunteers showed a reaction with the recycled paper preparation while no such reaction was seen upon the application of the conventional type of toilet paper. The validity of these findings was corroborated by corresponding results in the use test., Conclusion: There is clearly a potential for allergic reactions to components of moist toilet paper and reactions to recycled toilet paper presumably irritant by nature. These irritant reactions are probably caused by the rough texture of current paper types and do not reflect the presence of potentially toxic ingredients such as metal salts.

A controlled randomized study was performed in 60 patients with 64 chronic, dry tympanic membrane (TM) perforations. The perforations were randomly allocated to either resection of the perforation rim and instillation of 1% hyaluronan (Healon; HYA) in the perforation gap once daily for 7 days (33 ears) or resection of the perforation margin and application of a sterile rice paper prosthesis (31 ears). The treatment effect was documented by TM photography and morphometric measurements of the perforation area. The hearing was assessed with puretone and high-frequency audiometry. After 2 months, 5 of the HYA-treated perforations (15%) and 4 of the rice-paper-treated TMs (13%) were healed. After 1 year, 18 perforations (9 in each treatment group) were healed. In neither group were there any persistent adverse effects on hearing. It is noteworthy that 28% (18/64) of the chronic, long-standing TM perforations could be repaired by these technically simple and time-saving methods. Both procedures should be considered as easy first-choice alternatives to myringoplasty in selected cases.

The aim of the present study was to evaluate the significance of diffusion barriers in the development of artificial caries-like lesions. A 30-mm2 enamel surface area was covered with a layer of filter paper and exposed to a gently agitated 50 mM acetate buffer, pH 4.5, containing either 0.1 or 1.0 ppm fluoride for up to 48 h at 20 degrees C. Similar enamel surfaces, but unprotected by filter paper, served as controls. It was found that below the filter paper an approximately 60-microns-deep caries-like lesion was developed exhibiting a preserved surface layer, the mineral content of which depended on the fluoride concentration of the buffer. The lesions of the control teeth without a filter paper coverage were erosion-like surface lesions. In the filter paper calcium and phosphate released from the enamel combined with fluoride from the bulk solution had established a supersaturation with respect to fluorapatite while the undersaturation with respect to hydroxyapatite was maintained, which caused a caries lesion to develop. The lesions produced in noncovered enamel by bulk solution unsaturated with respect to both apatites were erosion-like. It was concluded that a diffusion barrier can significantly favor the establishment of supersaturation with respect to fluorapatite and thus facilitate the development of a subsurface caries-like lesion at the expense of the erosion.

Screening of infants for congenital adrenal hyperplasia (CAH) using filter paper blood samples collected on the 5th day of life was performed with a radioimmunoassay for 17-hydroxyprogesterone without extraction with organic solvents. A total of 153,000 newborns were screened and 12 cases of CAH were detected (1:12,800). With recall levels related to gestational age, the recall rate could be lowered to 0.05%.

Disposable diapers are now being used in incubators, raising questions concerning the amount of dust they produce during diaper change, and the subsequent effects on premature infants. This study compares the amount of dust produced during change of disposable diapers and during change of cloth diapers. The disposable diapers tested were Pampers (premature baby size, sterilized), manufactured by the Procter & Gamble Company. Cloth diapers tested included diaper covers. Results of the study indicated that disposable diapers produce far less dust than cloth diapers. Thus, disposable diapers can be satisfactorily used in incubators for premature infants. The difference in results between the two types of diapers might stem from their different structures: cloth diapers have a monolayer structure, whereas disposable diapers have a double cover over the refined pulp they contain, which might otherwise tend to generate dust.

An ELISA for the quantitation of the C5a anaphylatoxin of complement in human plasma is presented which is based on the use of both a monoclonal antibody and polyclonal antibodies to C5a. Its detection limit is 1 ng C5a/ml plasma. It detects no C5a in fresh EDTA-plasma. Complete removal of C5 by an optimized precipitation step prior to the assay procedure is an essential feature of the method.

In an experiment on skin rash caused by diapers, adult skin was used to compare the effects of disposable diapers (A; Pampers), conventional disposable diapers (B), and cotton diapers. Pieces of each type of diaper measuring 2 X 2 cm and containing 0.2 ml saline solution were pasted on forearm skin for 5 h. Water content of the corneum was measured with an impedance meter 1 h after the pieces were removed. Results were excellent for diapers A and B: there were no significant differences observed in water content of the corneum when A and B were compared with conventional cotton diapers. The advantages of disposable diapers were confirmed in infants.

The amount of active groups of cyanate esters and imidocarbamates present on cyanogen-bromide-activated paper discs were estimated. The protein-binding capacity of activated discs was found to be dependent on the amount of these active groups. The results suggested that monitoring of active groups on activated discs can help in preparing consistent allergosorbent for immunoassays.

Sera of 60 allergic persons and 20 healthy RAST-negative control persons were examined using immunoblotting to establish the presence of grass pollen-specific antibodies. These antibodies were depicted by an enzymatic, immunoglobulin class-specific indicator system. Seven bands of the timothy pollen extract proved to be major allergens. One of the 4 grass pollen-specific monoclonal antibodies tested (Bo 1) recognized the 7 major allergen bands. The possibility of employing monoclonal antibodies for the isolation of allergens is discussed.

Immunoglobulin E (IgE) was demonstrated by a double antibody radioimmunoassay technique (PRIST) in 5 of 17 unconcentrated fecal extracts from children. Four of the PRIST-positive extracts also had measurable levels of IgE determined by an enzyme-linked immunosorbent test (Enzygnost IgE). However, using a competitive antibody radioimmunoassay technique (RIST), IgE was found in all unconcentrated fecal extracts. The RIST-IgE levels of the extracts were higher and did not correlate to IgE measured by the other methods, nor to the IgE in serum from the same children or to the manifestation of allergy. On the other hand, 4 of the 5 children with measurable PRIST-IgE levels, and 3 of the 4 children with detectable Enzygnost IgE concentrations in the extracts, had elevated serum IgE as well as a history of allergy. Gel filtration studies indicated that IgE determined by the PRIST and Enzygnost IgE methods had been degraded to fragments of lower molecular weight than that of albumin. This study also suggests that the IgE in feces measured by the RIST method is overestimated due to the influence from nonspecific substances of high molecular weight.

The performance of ELISA to detect IgG and IgM antibodies to Aspergillus fumigatus has been evaluated in strongly precipitin-positive, weakly precipitin-positive and precipitin-negative patient sera, with immunoblot analysis as the confirmatory test. All strongly precipitin-positive sera contained increased IgG titers and showed clearly positive immunoblot patterns. Most of the weakly precipitin-positive sera contained ELISA titers within the normal range established with sera of healthy blood donors and showed normal immunoblot patterns. Increased titers of IgG and/or IgM were measured in one-sixth of the precipitin-negative patient sera. Immunoblot analysis confirmed the presence of antibodies to A. fumigatus in 55% of the precipitin-negative sera with increased antibody titers. ELISAs for A. fumigatus-specific IgG and IgM are sensitive tests for screening of patient sera. However, positive results with ELISA should be confirmed by means of immunoblot analysis.

Polyacrylamide gel electrophoresis in sodium dodecyl sulphate (SDS) followed by protein staining has shown that extracts from 11 different grass pollens contained proteins with similar molecular weights to that of the allergen R7 from rye grass (Lolium perenne) pollen extract (i.e. 29,000-31,000 daltons). Western blotting and detection with polyclonal (rabbit) antibodies raised against the purified R7 (Rye I) allergen indicated that these proteins were antigenically related and their allergenic properties were demonstrated by the binding of human IgE to immunoblots. The distribution of cross-reacting antigenic determinants was further investigated by immunoblotting with 2 mouse monoclonal antibodies, R7M1 and R7M2, produced with purified R7 as the initial immunogen. The 2 monoclonal antibodies were shown to react with 'R7-like' components of grass pollen extracts other than the component from rye grass. Differences in the distribution of R7M1 and R7M2 binding were found indicating that they are directed at separate R7 epitopes.

The time course of the appearance of myelin-specific markers was studied in the developing chick central nervous system (CNS). Chick CNS tissue was studied for the presence of both proteolipid and myelin basic protein by electroblotting and for 2',3'-cyclic nucleotide 3'-phosphohydrolase (CNPase) by enzyme assay. Four regions of chick spinal cord (cervical, brachial, thoracic and lumbar), brain stem, cerebellum, optic nerve and cortex were studied. In general, myelin basic protein appeared approximately 1 day earlier than proteolipid. In spinal cord and brain stem, myelin basic protein appeared at 13 days incubation. In cerebellum and optic nerve, it appeared at 17 days incubation and in cortex at hatching. CNPase activity increased in most CNS regions between 16 days incubation and hatching. These results suggest that myelination occurs earlier in the chick than in the rat and that it occurs over a shorter time period.

Monoclonal antibodies against major allergens of the short ragweed pollen were produced by fusion of NS-1 cells with splenic cells from BaLB/C mice that had been immunized separately with the major allergens, AgE-B + E-C and AgK, of the short ragweed pollen. These monoclonal antibodies were detected by enzyme-linked immunosorbent assay and further characterized by immunoblot analysis using the crude extract and highly purified allergens of the ragweed pollen. Three monoclonal antibodies obtained by immunization with AgE-B + E-C, designated as 36-6, 27-2 and 48-5, reacted mainly with the beta (36-6) and alpha (27-2) subunit of AgE and both AgE and AgK (48-5), respectively. Two monoclonal antibodies obtained by immunization with AgK, 4-7 and 8-5, had a similar reactivity with AgE-B + E-C, AgK, AgK-A and AgK-B. In addition, however, antibody 4-7 also reacted with AgE-B2 as well as the 36- and the 24- to 22-kilodalton antigens of the crude extract. All 5 monoclonal antibodies were characterized as IgGl subclass. Besides, monoclonal antibody 48-5 also showed cross-reactivity to components of sage pollen. Beyond academic interests, the monoclonal antibodies described here may also be useful in clinical allergy.

Antigliadin antibodies (AGA) were analyzed by immunoblotting from the sera of 30 patients with dermatitis herpetiformis (DH) and of 13 patients with gluten-sensitive enteropathy (GSE). The results were correlated to the jejunal morphology and to the serum AGA values as quantified with an enzyme-linked immunosorbent assay (ELISA). Immunoblotting analysis disclosed a distinct pattern of AGA which was similar in patients with DH and GSE. In both diseases individual variation in IgG class AGA patterns was large, suggesting that several antigenic determinants are involved in the AGA response. The IgA class AGA pattern was clearly more homogeneous. The sera with several bands in immunoblotting had the highest AGA levels in ELISA. Strong staining in the main gliadin area (bands from 31 to 38 kd) indicated severely damaged intestinal mucosa whereas the serum of DH patients with partial villous atrophy or normal mucosa showed usually weaker staining. The immunoblotting pattern of sera from healthy controls was even weaker but was directed against the same polypeptides as in the patients suggesting a quantitative rather than qualitative difference between healthy and diseased subjects. This difference was, however, more obvious in the IgA than in the IgG class AGA pattern possibly indicating a more fundamental defect in the regulation of synthesis of IgA isotype in DH and GSE.

A review of the history of diaper use in various countries reveals a number of difficulties that have been confronted. These difficulties seem to be associated with both climatic and traditional factors in each country. In Japan, for example, because of the hot and humid summers, there was a need to make diapers both leak- and 'stuffiness'-proof. At first the use of natural fiber diaper covers met this need, but the recent development of disposable diapers seems to even better solve these problems. Yet disposable diapers still make up only 25% of the diaper market in Japan. This is because the Japanese view of baby care and the country's economy have worked against acceptance of disposable diapers, despite their merits. The slower trend toward disposable diaper use in Japan is in contrast to their rapid acceptance in Western Europe and the United States. Western acceptance probably stems from the perception that disposable diapers are a convenient and practical alternative to cotton diapers. Japan is moving in the direction of greater acceptance, but will do so according to Japanese customs and thought.

Parietaria judaica pollen allergens, fractionated by SDS-polyacrylamide gel electrophoresis and transferred to nitrocellulose membranes, were identified using 52 sera collected in Australia and Sicily from P. judaica pollen-allergic patients. IgE-binding pollen components transferred to nitrocellulose were detected by reaction with 125I-anti-human IgE and autoradiography. Nine pollen components, ranging in molecular weight (MW) from approximately 10,000 to 80,000 daltons, bound IgE antibodies but the two fastest migrating components sometimes each separated into two very closely migrating bands. The faster of the two components exhibiting doublet formation (MW approximately 10,000 daltons) showed by far the highest frequency of IgE binding, being recognised by 50 of the 52 sera examined. Although patients' IgE reaction patterns to P. judaica allergens were heterogeneous, the degree of heterogeneity was much less than that observed with house dust mite and other pollen extracts studied by electrophoretic transfer analysis. Results with gradient gel-nitrocellulose transfer experiments which showed no IgE-binding components with MWs less than 70,000 daltons, and comparisons of our electroblotting results with crossed radioimmunoelectrophoresis results of others, suggested that the doublet proteins with MWs of approximately 10,000 probably bind to higher MW proteins in P. judaica pollen extracts.

The allergen composition of moulds (Alternaria alternata and Aspergillus fumigatus) and mites (Dermatophagoides pteronyssinus and Dermatophagoides farinae) was studied by electroblotting. The allergens were separated by SDS-gPAGE and transferred to a nitrocellulose membrane by electroblotting. Specific IgE antibodies directed against mould and mite allergens were utilized as probes to detect the allergens. The bands were localized on the nitrocellulose membrane by means of enzyme- or isotope-labelled antibody. Circulating specific IgE antibodies in mould and mite sensitive patient sera were also analyzed and characterized with respect to their IgE specificity. Under appropriate experimental conditions this electroblotting technique could also be used for quantitation of individual allergens by scanning the isotope- or enzyme-stained nitrocellulose strips.

Some workers when using cyanogen bromide activated paper as a solid immunoabsorbent have reported variable success in radio immuno assays. A quantitative investigation has been made of some of the factors which effect the activation of various papers with CNBr, and also of the subsequent coupling reaction with the sensitizing antigen. Different papers have been found to react in different ways, and some papers take up so little antigen that they are unsuitable for use for this purpose.