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PubMed1976

Microaggregate formation in stored human packed cells: comparison with formation in stored whole blood and a method for their removal.

Experiments were performed to compare the formation of microaggregates in stored human whole blood (WB) with that in stored packed cells (PC) and also to compare the effectiveness of standard blood transfusion filters with dacron wool (Swank) micropore transfusion filters in removing such microaggregates. After 5, 10, 15 and 20 days of storage SFP and debris weights of PC's were considerably greater than those of matched WB samples. Passage of either WB or PC's through standard blood transfusion filters resulted in small decreases in SFP and debris weights. Passage of either WB or PC's through dacron wool (Swank) transfusion filters led to striking and highly significant decreases in both SFP and debris weights. When stored PC's were diluted to the same hematocrits as their corresponding WB samples, SFP remained considerably elevated above those of the WB samples. On the basis of this research, it is concluded that centrifugation of blood during component separation leads to a significant increase in microaggregate formation over and above that which progressively occurs during storage and that the risk of pulmonary microembolization during transfusion with stored PC's is greater than that during WB transfusion. For this reason, dacron wool (Swank) filters should always be used when PC's are being transfused.

Blood Preservation↗
PubMed2001

Historical cohort study of US man-made vitreous fiber production workers: II. Mortality from mesothelioma.

As part of our ongoing mortality surveillance program for the US man-made vitreous fiber (MMVF) industry, we examined mortality from malignant mesothelioma using data from our 1989 follow-up of 3478 rock/slag wool workers and our 1992 follow-up of 32,110 fiberglass workers. A manual search of death certificates for 1011 rock/slag wool workers and 9060 fiberglass workers revealed only 10 death certificates with any mention of the word "mesothelioma." A subsequent review of medical records and pathology specimens for 3 of the 10 workers deemed two deaths as definitely not due to mesothelioma and one as having a 50% chance of being caused by mesothelioma. Two other deaths, for which only medical records were available, were given less than a 50% chance of being due to mesothelioma. Eight of the 10 decedents had potential occupational asbestos exposure inside or outside the MMVF industry. We also estimated the mortality risk from malignant mesothelioma in the cohort using two cause-of-death categorizations that included both malignant and benign coding rubrics. Using the more comprehensive scheme, we observed overall deficits in deaths among the total cohort and fiberglass workers and an overall excess among rock/slag wool workers. The excess in respiratory system cancer is largely a reflection of elevated lung cancer risks that we attributed mainly to confounding by smoking, to exposures outside the MMVF industry to agents such as asbestos, or to one or more of the several co-exposures present in many of the study plants (including asbestos). The second scheme, which focused on pleural mesothelioma in time periods when specific malignant mesothelioma coding rubrics were available, classified only one cohort death as being caused by malignant mesothelioma, compared with 2.19 expected deaths (local county comparison). We conclude that the overall mortality risk from malignant mesothelioma does not seem to be elevated in the US MMVF cohort.

Adult↗
PubMed2001

Historical cohort study of US man-made vitreous fiber production workers: V. Tobacco-smoking habits.

As part of our ongoing mortality surveillance program for the US man-made vitreous fiber industry, we surveyed a random sample of study members to estimate tobacco-smoking habits for the total cohort. Separate sampling frames were constructed for four study groups: male and female workers within the fiberglass and rock/slag wool subcohorts. The frames included all persons who had worked a year or more between 1945 and 1986 (with some exceptions), and who were alive as adults (18+ years) on January 1, 1980, the year the age distribution of the cohort most resembled the US comparison population. Subjects were randomly selected from the frames, and a structured telephone interview was administered to the subject or a proxy respondent between January 1995 and December 1997. Using survey data, we estimated the point prevalence of ever and current cigarette smoking on January 1, 1980, and made comparisons with other occupational groups and general populations. Overall response rates (interviews/targeted sample) were greater than 78% for each of the four study groups. From our estimates, we infer that male workers from both the fiberglass and rock/slag wool cohorts and female rock/slag wool workers had higher rates of ever smoking than the corresponding general populations of the United States and most of the states where the study plants were located. These findings suggest that at least part of the elevated externally standardized mortality ratios (US and regional rate-based) for respiratory system cancer noted among male subjects and the male-dominated total cohort in our previous cohort analyses were due to uncontrolled positive confounding by smoking.

Adolescent↗
PubMed1997

Peptidylarginine deiminase of the hair follicle: characterization, localization, and function in keratinizing tissues.

The enzyme peptidylarginine deiminase (PAD; EC 3.5.3.15) is responsible for the formation of protein-bound citrulline, a major amino acid in the inner root sheath (IRS) and in the medulla of the hair follicle. From mainly biochemical evidence, it is known that the substrate for the enzyme is trichohyalin and that trichohyalin granules gradually disappear to form a matrix with intermediate-like filaments in the IRS cells. In the medulla, the granules aggregate into large masses without filaments. The proteins in both the IRS and medulla are finally cross-linked by transglutaminase. A corollary of the apparent central role of PAD acting on the trichohyalin protein in these processes is that it should be present in the IRS and medulla cells, coincident with trichohyalin. Hair-follicle PAD has not previously been isolated. In the current study, the enzyme was isolated from wool follicles of adult sheep and peptide sequences were used to design DNA primers for the synthesis of PCR products from follicle mRNA. Subsequently, a PAD-specific complementary RNA probe and a trichohyalin complementary RNA probe were prepared for localization studies by in situ hybridization in wool follicles and the epithelia of the rumen, embryonic hoof, and tongue papillae. The experiments have revealed a striking co-expression of PAD and trichohyalin in all of these tissues. The amino acid sequence of the wool-follicle PAD molecule has been deduced from sequencing of the cloned PCR products.

Amino Acid Sequence↗
PubMed1988

Cloning and characterization of a mouse type I hair keratin cDNA.

A cDNA library was prepared from poly(A)-containing C57BL/6J mouse hair-root-enriched mRNA. The library was screened using a radiolabeled nucleic acid probe prepared from a sheep wool, Type I keratin cDNA clone (SWK2). Clone MHKA-1 was shown to contain a mouse hair, Type I keratin cDNA insert by positive hybridization-selection translation assay, and by the corresponding deduced amino acid sequence. The size of the cDNA insert is 1585 bp (excluding homopolymer tails) and on the basis of Northern blot analysis it corresponds to a mRNA of approximately 1.6 Kb. The cloned cDNA sequence includes the entire coding region for a protein of 416 amino acids (including the initiation methionine). Comparison of the deduced amino acid sequence with that of sheep wool, Type I keratin (8c1) reveals an overall corresponding sequence identity of 87%. In contrast, the MHKA-1 protein is significantly less similar to non-hair Type I keratins. An additional 3 amino acids (adjacent proline residues) not present in the wool protein have been identified near the middle of the carboxy-terminal end of the mouse protein. MHKA-1 is the first of a series of mouse hair follicle cDNA clones to be identified and characterized that will enable us to study follicular regulatory mechanisms and the interrelationships among the proteins in the mammalian hair follicle.

Amino Acid Sequence↗
PubMed1987

Low molecular weight factors displaying augmenting activity for human antibody production in vitro.

Dialyzable low molecular weight antibody-augmenting factors (LMAAF) were found in the culture supernatant of human tonsillar lymphocytes which were not stimulated by antigen and/or mitogen in vitro. Phagocyte-depleted nylon wool-adherent lymphocytes (M-Ny+ cells) were responsible for the release of the LMAAF. Marbrook's culture system was adopted to assay for the LMAAF. The M-Ny+ cells, which were cultured without antigen and/or without mitogen in the reservoir of Marbrook's diffusion culture vessel, released the LMAAF, which diffused across a dialysis membrane and significantly augmented the pokeweed mitogen (PWM)-induced plaque-forming cell (PFC) response of phagocyte-depleted lymphocytes (M-cells) cultured in the inner vessel. Phagocyte-depleted nylon wool-passed lymphocytes (M-Ny- cells) cultured in the reservoir could not augment the PWM-induced PFC response of the M- cells cultured in the inner vessel. The exuded fluid, which was the dialysate of the culture supernatant of the M-Ny+ cells ultrafiltrated with dialysis tubing, also enhanced the PFC response of M- cells cultured in 24-well multi plates. The exuded fluid also augmented the total IgM and IgG production of human tonsillar and peripheral blood lymphocytes measured by enzyme-linked immunosorbent assay (ELISA) systems. Gel filtration chromatography on Sephadex G-25 Superfine column showed that the LMAAF activity was demonstrated in the fractions corresponding to a molecular weight (m.w.) of 362 to 1,355 and a m.w. of 3,560 to 5,700, with a peak activity at about 4,500 dalton. The LMAAF were inactivated by treatment with proteinase K, but not by trypsin, alpha-chymotrypsin, RNase, and DNase, and were stable when treated at 56 C for 60 min. The dialysates of culture supernatants from two out of seven Epstein-Barr virus (EBV)-transformed M-Ny+ cell lines showed LMAAF-like activity. These results indicate that phagocyte-depleted nylon wool-adherent lymphocytes, possibly B cells, release low molecular weight factors displaying augmenting activity for human antibody production in vitro.

Antibody Formation↗
PubMed1991

Immunological tolerance induced by liver grafting in the rat: splenic macrophages and T cells mediate distinct phases of immunosuppressive activity.

In the rat combination DA into PVG, liver grafts are not rejected but induce donor-specific transplantation tolerance. We have examined the immunosuppressive properties of spleen cells from PVG recipients of DA liver grafts at various times post-grafting. The results indicate the development of two phases of cell-mediated suppressor activity, which appear to be mediated by separate spleen cell populations. Mitomycin-C-treated spleen cells taken from animals between 5 and 28 days post-grafting were able to suppress rat mixed lymphocyte reactions (MLRs). These 'early' suppressor cells were glass adherent and absent from populations purified by passage through nylon wool or G10 Sephadex columns. Suppression of MLR by purified glass adherent cells was not specific for either stimulator or responder haplotypes and was blocked by indomethacin. Nylon wool purified T cells were not suppressive at this time. Spleen cell suppressor activity declined to background levels after 35 days post-grafting. However, spleen cells from long-term surviving liver graft recipients (20 weeks or more) were again able to suppress MLR; the 'late' suppressor cells were nylon wool non-adherent and suppression was specific for the donor (DA) MHC type. We conclude that liver grafting in this combination generates early and late phases of suppression among spleen cells, that the early phase is produced by macrophages and mediated by prostaglandins and that the late phase is dependent on allospecific suppressor T cells.

Animals↗
PubMed1981

Ia antigens on antigen-presenting cells do not carry the same Ia specificities as detected on suppressor and helper T cells.

The passage of spleen cells through a tightly packed nylon wool column partially separated two populations of cells required for the antigen-induced secondary proliferative response of T cells. The proliferating T cells were in the nylon wool column-passed cell population, but the addition of a small number of nylon-wool-adherent cells was required for their maximal proliferative response. Such adherent cells were non-T, non-B, Ia antigen-positive cells serving as antigen-presenting cells. Ia determinants on this cell type are encoded in two separate I subregions, I-A and I-E/C, and they are expressed simultaneously on the same cells. Antisera specific for I-J subregion gene products having as a known activity the killing of antigen-specific suppressor and helper T cells were unable to kill antigen-presenting adherent cells. The results indicated that at least I-J subregion gene products expressed on suppressor and helper T cells are not present on antigen-presenting accessory cells. The Ia specificities detected on accessory cells follow the cross-reactive pattern found in Ia molecules of B cells, and thus antigen-presenting cells carry Ia antigen identical to those of B cells, and not those of T cells.

Animals↗
PubMed1998

Sperm separation in patients with urogenital infections.

A total of 196 patients attending the Center of Dermatology and Andrology, Giessen, Germany, were examined for fertility problems. Polymorphonuclear elastase, reactive oxygen species (ROS) and the number of round and peroxidase-positive cells were investigated in addition to routine semen analysis. The ejaculates were also analysed before and after sperm separation by means of swim-up or glass wool filtration. In 20 cases of leukocytospermia, sperm concentration, motility, viability, production of reactive oxygen species, and the number of peroxidase-positive cells were evaluated before and after glass wool filtration. The results show that ROS production by viable spermatozoa is highly correlated with the concentration of PMN elastase and the number of both peroxidase-positive and round cells. Multiple regression analysis with motility as dependent parameter showed the number of round cells (n = 91; r = -0.332; P = 0.0030) to be the most important parameter affecting motility, while ROS mainly affects the viability of spermatozoa (n = 69; r = 0.250; P = 0.0107). In the case of leukocytospermia, glass wool filtration significantly reduced the number of peroxidase-positive cells and ROS production (P = 0.0098 and P = 0.0005, respectively). Receiver operating characteristic (ROC) curve analysis for ROS production in the ejaculate using a concentration of 1.000 ng ml-1 PMN elastase as decisive parameter resulted in a cut-off value of 49,489.9 counts 10(-7) viable spermatozoa. The statistical parameters were: Sensitivity: 63.2%, specificity: 100%, positive predictive value: 100%, negative predictive value: 36.1%.

Cell Separation↗
PubMed2005

Systemic associations of retinal microvascular signs: a review of recent population-based studies.

Retinal microvascular signs, such as generalized retinal arteriolar narrowing, focal arteriolar narrowing, arteriovenous nicking and retinal haemorrhages, microaneurysms and cotton wool spots, are common fundus findings in the general population, even in individuals without hypertension or diabetes. Recent population-based studies have provided new insights into the systemic associations and clinical significance of these retinal signs. Studies show that these retinal microvascular signs are strongly associated with elevated blood pressure (BP). Generalized retinal arteriolar narrowing may be associated with markers of inflammation and risk of diabetes and hypertension. Retinal haemorrhages, microaneurysms and cotton wool spots are associated with risk of subclinical and clinical stroke, cognitive impairment, renal dysfunction and cardiovascular mortality, independent of BP and cardiovascular risk factors. A consistent pattern of association between retinal microvascular signs and ischaemic heart disease has not been demonstrated. This suggests that patients with some retinopathy signs (retinal haemorrhages, microaneurysms and cotton wool spots) may benefit from a careful systemic evaluation and, if supported by further research, appropriate risk reduction therapy.

Aneurysm↗
PubMed1994

Rubbing test responses of the skin to man-made mineral fibres of different diameters.

The irritant potency of 5 types of insulation wools (2 types of rockwool and 3 types of glasswool) were studied on the basis of their differences in diameter by a standardized rubbing test. Assessment was done by laser Doppler flowmetry (LDF) and erythema scores. Insulation wools show a wide distribution of the diameter around the mean. Of the rockwool material, 30% consisted of small grains, called "shot". The control test site did not change over time; thus, the rubbing procedure itself does not seem to contribute to the skin reactions. All fibre types proved to be irritant to the skin. The irritant reactions did not correspond with the mean diameter, although the fibre type with the smallest diameter was the least irritant and the fibre type with the largest diameter the strongest. The presence of "shot" had a significant effect on the LDF responses. "Shot" and the distribution around the mean diameter may play a role in eliciting the skin irritation by insulation wools.

Adult↗
PubMed1993

Hypertonic saline test for ophthalmic nerve impairment.

In order to compare the accuracy of methods for testing ocular surface sensitivity (e.g. function of the first branch of the trigeminal nerve) three different methods were compared in patients with unilateral acoustic neurinomas. The three methods 1) hypertonic (3%) saline in the conjunctival sac (apparently not previously described in the literature) 2) esthesiometer (Cochet & Bonnet) touching of the cornea, and 3) touching the cornea with a cotton wool wisp, was found to reveal reduced ocular surface sensitivity on the neurinoma side in 50% (hypertonic saline), 23% (esthesiometer) and 14% (cotton wool wisp) of cases, respectively. With McNemar's test for comparing test methods the 3% saline test proved significantly more sensitive than the cotton wool wisp test (p < 0.05), but not significantly more sensitive than the esthesiometer test (p > 0.10). The advantage of the 3% saline test, apart from its high sensitivity, is that it does not require sterilization of any equipment as is the case for the esthesiometer nor, in contrast to the methods using corneal or conjunctival touch, does it require perfect visual control by means of a magnifying glass in order to be performed accurately, and it is not affected by visual stimuli. It should therefore be the preferred test of assymetry in ophthalmic nerve function.

Cornea↗
PubMed1993

Clinicopathology of a case with retinopathy of pancreatitis.

Bilateral retinopathy of pancreatitis developed in a 36-year-old male patient during recurrent acute necrotizing pancreatitis. The retinopathy remained clinically unchanged between the detection and the death 6 days later. Characteristics of the retinal lesions were histologically similar to cotton-wool spots of other origin, but all the retinal vessels including vessels around and in the cotton-wool spots were intact and contained plasma and red blood cells. Absence of vascular occlusion and endothelial damage suggests that in acute pancreatitis vascular embolisation in the retina is not necessary to the development of cotton-wool patches.

Acute Disease↗
PubMed2000

An alpha-proteobacterium converts linear alkylbenzenesulfonate surfactants into sulfophenylcarboxylates and linear alkyldiphenyletherdisulfonate surfactants into sulfodiphenylethercarboxylates.

The surfactant linear alkylbenzenesulfonate (LAS; 0.5 mM) or linear monoalkyldiphenyletherdisulfonate (LADPEDS; 0.5 mM) in salts medium was easily degraded in laboratory trickling filters, whereas carbon-limited, aerobic enrichment cultures in suspended culture with the same inocula did not grow. We took portions of the trickling filters which degraded LADPEDS, shook the organisms from the solid support (polyester), and found that growth in suspended culture in LADPEDS-salts medium occurred only in the presence of some solid support (polyester fleece or glass wool), though little biomass was immobilized on the support. The end products in suspended culture were identical with those from the trickling filters. There was low plating efficiency of LADPEDS-grown cultures on complex medium, and no picked colony or mixture of colonies grew in LADPEDS-salts-glass wool medium. However, selective plates containing LADPEDS-salts medium solidified with agarose yielded LADPEDS-dependent, pinpoint colonies which could be picked singly and subcultured in selective liquid medium. Isolate DS-1 was a bacterium which showed 93% sequence homology (16S ribosomal DNA) to its nearest phylogenetic neighbor, an alpha-proteobacterium. Strain DS-1 grew heterotrophically in LADPEDS-salts-glass wool medium and converted the set of aryl-substituted alkanes to the corresponding aryl-substituted carboxylic acids of shorter chain length. Similarly, strain DS-1 grew heterotrophically with commercial LAS, converting it to a set of sulfophenylcarboxylates. Growth with a single isomer of LAS [3-(4-sulfophenyl)dodecane] was concomitant with excretion of 4-(4-sulfophenyl)hexanoate, which was identified by matrix-assisted laser desorption ionization mass spectrometry. The growth yield (6.4 g of protein/mol of C) indicated mass balance, which, with the specific growth rate (0.05 h(-1)), indicated a specific utilization rate of LAS of 2.2 mkat/kg of protein.

Alphaproteobacteria↗
PubMed1966

Evaluation of filters for removal of bacteriophages from air.

Glass wool, nonabsorbent cotton, fiberglass filter medium, and a commercial absolute filter were tested for effectiveness in removing aerosolized bacterial viruses under low flow rate (1 ft(3)/min) and high flow rate (10 to 25 ft(3)/min) air-flow conditions. Special equipment was designed for measurement of filter efficiencies under the two air-flow conditions. Under low air-flow rate test conditions, glass wool was only 98.543 to 99.83% efficient, whereas cotton (five layers), fiberglass medium (three layers), and the commercial absolute filter were at least 99.900, 99.999, and 99.999 efficient, respectively. Glass wool and cotton were not used under higher air-flow conditions because they were difficult to assemble in leak-tight filters. The commercial absolute filter and fiberglass medium (three layers) were at least 99.990 and 99.999% efficient, respectively, under the higher air flow conditions. A stainless-steel filter of simple design and fitted with three layers of fiberglass medium was found to be greater than 99.999% efficient in removing high concentrations (20,000 to 70,000 plaque-forming units per cubic foot) of aerosolized bacteriophages from air moving at a low flow rate (1 ft(3)/min). Use of this filter on pressure-vacuum tanks in the fermentation industry is suggested. Several other uses of such a filter are proposed.

Air Microbiology↗
PubMed1983

Pharmacological modulation of suppressor cell activity in mice with disseminated histoplasmosis.

Indomethacin and cyclophosphamide (CY) were used in an attempt to modify the suppressive effects of spleen cell populations from mice with disseminated histoplasmosis at 1 week of infection. In vitro addition of indomethacin did not alter the depressed plaque-forming cell response to sheep erythrocytes of normal spleen cells cocultured with unfractionated or nylon wool-fractionated spleen cells from infected mice. Likewise, indomethacin given intraperitoneally did not enhance the subnormal in vivo plaque-forming cell response of spleen cells from infected mice. Conversely, 20 mg of CY per kg given intraperitoneally 2 days before or 6 h after the inoculation with Histoplasma capsulatum partially reversed the suppression effected by splenic T cells (nylon wool passed) in vitro, whereas 50 mg of CY per kg given intraperitoneally 6 h after the injection of H. capsulatum ablated suppressor T cell activity in vitro; neither dosage of CY altered the suppression mediated by unseparated or nylon wool-adherent spleen cells. Furthermore, the administration of 50 mg of CY per kg failed to improve the depressed footpad responses of mice infected for 1 week to sheep erythrocytes in sheep erythrocyte-sensitized mice or to histoplasmin. These findings indicate that in experimental disseminated histoplasmosis, suppression effected by splenic T cells can be alleviated by CY; however, there is a persistent immunosuppressor mechanism(s) that cannot be counteracted by either indomethacin or CY.

Animals↗
PubMed1984

In vitro effects of natural killer cells against Paracoccidioides brasiliensis yeast phase.

Recently, data have been reported suggesting natural killer (NK) cells may function in natural resistance against a fungus, Cryptococcus neoformans. The primary objective of this study was to examine the reactivity of murine splenic cells against another fungus, Paracoccidioides brasiliensis. Levels of NK activity in effector cell pools were varied by: (i) removing nylon wool-adherent cells, (ii) fractionating splenic cells on Percoll discontinuous gradients, (iii) using old and young effector cell donor mice, (iv) using donors from different strains, and (v) pretreating donors with NK-augmenting and -depressing agents. The various effector cell pools were simultaneously used in the 4-h 51Cr release assay with YAC-1 targets to determine the NK reactivity and in the in vitro growth inhibition assay against P. brasiliensis yeast phase targets. In each case, the level of NK reactivity correlated with the ability of the effector cells to inhibit the in vitro growth of P. brasiliensis. NK activity and P. brasiliensis growth-inhibiting ability could be augmented by fractionation of splenic cells through nylon wool or Percoll gradients. The effector cells responsible for the NK activity and P. brasiliensis growth inhibition were characterized as being nylon wool nonadherent, being found in the low-density fractions from Percoll discontinuous gradients, and having no detectable Thy-1 antigen or immunoglobulin but having asialo GM1 on their surface. These data support the contention that NK or NK-like cells are responsible for limiting the in vitro growth of P. brasiliensis.

Animals↗
PubMed1985

Blastogenic response of bovine lymphocytes to Brucella abortus lipopolysaccharide.

Brucella abortus lipopolysaccharide was tested in a blastogenesis assay with unfractionated and nylon wool-separated peripheral blood lymphocytes of Brucella-naive cattle and cattle immunized with B. abortus. Our results indicated that in cattle the lipopolysaccharide of B. abortus is not a B-cell mitogen. In immunized animals it stimulated predominantly nylon wool-adherent cells. The lipopolysaccharide of Escherichia coli O128:B12, in contrast, induced a substantially greater proliferative response in circulating lymphocytes, predominantly those adherent to nylon wool, of the Brucella-naive cattle.

Animals↗