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Biomedical subjects

S A Little

Publications and source records attributed to S A Little.

At least 37 records · Page 2Linked to original sources
PubMed1996

Improved diagnosis of allergic bronchopulmonary aspergillosis with gp66 (formerly antigen 7) of Aspergillus fumigatus for specific IgE detection.

BACKGROUND: In cystic fibrosis (CF) serologic tests assume increased significance for discriminating the symptoms of allergic bronchopulmonary aspergillosis (ABPA) from those of bacterial infections. Elevated specific IgE is a sensitive indicator of ABPA, but the high prevalence of patients with CF who have simple allergy to Aspergillus fumigatus means that it lacks specificity. OBJECTIVE: The aim of this study was to develop an assay to detect IgE to a single allergenic protein of A. fumigatus to increase the discrimination between ABPA and uncomplicated A. fumigatus allergy. METHODS: to identify a more selective allergen, serum samples from nine patients with ABPA and five patients with allergy were used to screen A. fumigatus proteins for allergenic activity by crossed radioimmunoelectrophoresis. Molecular weights were determined by electrophoresis and immunoblotting, and carbohydrate content was analyzed by concanavalin A chromatography and endoglycosidase digestion. RESULTS: The allergen identified was found to be a glycoprotein with a molecular weight of 66 kd, referred to as gp66. An ELISA was developed to measure specific IgE to gp66, and the efficacy was compared with IgE detection by using the unfractionated A. fumigatus extract on serum samples from 14 patients with CF and ABPA, 42 patients with CF and allergy but not ABPA, and 23 patients with CF without ABPA or allergy. CONCLUSION: Use of gp66 increased the specificity of IgE analysis from 58.5% to 78.5%, whereas the sensitivity of 92.9% was maintained.

Allergens↗
PubMed1995

Clusterin expression during programmed and teratogen-induced cell death in the postimplantation rat embryo.

Clusterin appears to play a role in multiple cellular processes including reproductive cell function, lipid transport, complement regulation, and endocrine secretion. In addition, clusterin has been shown to be associated with both developmental and induced cell death. We have used immunohistochemistry and in situ hybridization to study the relationship between clusterin expression, normal programmed cell death (PCD) in the developing rat limb bud, and abnormal cell death induced by hyperthermia in day 11 rat embryos. Immunohistochemical localization of clusterin in day 14-16 limb buds showed that the most intense immunostaining was associated with the condensing mesenchyme of the developing digit, a tissue exhibiting low levels of PCD. Moreover, areas of digital cell death, confined to future interphalangeal spaces, were devoid of clusterin immunostaining. Clusterin immunostaining was also observed in the interdigital mesenchyme and partially overlapped the cell death that occurs in this tissue during the early development of the digits. Although clusterin immunostaining overlaps areas of interdigital cell death, most apoptotic cells in the interdigital mesenchyme and underlying the surface ectoderm were not associated with clusterin immunostaining. We also examined the expression of clusterin in day 11 rat embryos exposed to 43 degrees C, an exposure that induces extensive cell death primarily in the developing neuroepithelium. In control embryos cultured at 37 degrees C, clusterin mRNA and protein were expressed at high levels in the heart, a tissue that is completely resistant to the cytotoxic effects of hyperthermia. Within 2.5 hr after an exposure of 43 degrees C, clusterin mRNA showed a dramatic induction in the prosencephalic mesenchyme and only a modest induction in the prosencephalic neuroepithelium.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Composition in situ and in vitro of vascular smooth muscle laminin in the rat.

Vascular smooth muscle cells are surrounded by a basal lamina containing an array of macromolecules: included among these are the laminins, a family of oligomeric glycoproteins composed of subunits encoded by different genes. In this study, we have used monoclonal antibodies to several of these subunits, including S-laminin, laminin B2, and laminin B1, to study these proteins in tail artery, superior mesenteric artery, and aorta of rats. In situ, immunostaining for the B2 and S chains was present in the basal lamina, between the smooth muscle cells, throughout the tunica media. In contrast, B1 chain immunostaining was concentrated around cells in the inner media. To investigate whether smooth muscle cells can produce S-laminin, laminin B2, and laminin B1, smooth muscle cells from the superior mesenteric artery were grown in culture and laminin subunit expression determined. In early culture (4 days), immunostaining showed abundant laminin B2 and less B1 synthesis and incorporation into the matrix. Staining for S-laminin was even less intense than for B1 and was localized to areas where cells were densely packed. The same pattern of S-laminin immunostaining was seen during early culture in cells grown on fibronectin, type IV collagen, or gelatin. Immunoblotting detected S-laminin in the conditioned medium from early cultured cells. In later culture (12 days), S-laminin incorporation into the matrix increased markedly compared to incorporation at 4 days. At this time, cells are much more densely packed and multilayered with extensive matrix accumulation. Cyclical stretching of cells in vitro did not increase immunostaining for S-laminin. Together these data show that S-laminin is a component of the arterial media in situ and that in vitro S-laminin is synthesized by smooth muscle cells. Increased incorporation of S-laminin into the matrix in later culture correlates with the presence of a more extensive matrix, suggesting that matrix organization may be critical to S-laminin incorporation.

Animals↗
PubMed1995

Soft lens movement: effect of blink rate on lens settling.

Little is understood about the mechanism by which soft lenses settle on the eye, although it has been suggested that lens base curve, steepening with in-eye lens dehydration or tear film changes may influence lens movement in the initial period of wear. In this study, we investigated the role of postlens tear fluid expulsion by assessing lens movement in 20 subjects wearing 38% water content hydrogel lenses for 10 min under three different conditions: 10 blinks/min, 30 blinks/min and eye closure. Over the 10 min wearing period, the total decrease in median lens movement for the 10 blinks/min condition was 0.07 mm, which was not a significant change (Friedman ANOVA, p = 0.13), while significant decreases occurred with conditions of 30 blinks/min (0.19 mm, p = 0.004) and eye closure (0.43 mm, p = 0.0001). As expected, lens movement under the three conditions was the same at insertion, but was significantly higher thereafter for the slower blink rate condition compared to the faster blink rate or eye closure conditions (Wilcoxon test, Z = 2.8 and -3.0, p = 0.006 and 0.003, respectively). Based on these findings, we postulate the model that the extent of lens settling and the degree of postinsertion lens movement are determined by the timeaverage pressure for postlens tear film expulsion exerted on the lens by the eyelids.

Adult↗
PubMed1995

Role of the post-lens tear film in the mechanism of inferior arcuate staining with ultrathin hydrogel lenses.

We studied the depletion of the post-lens tear film as contributing to inferior arcuate staining with ultrathin high water content hydrogel lenses. We monitored the post-lens tear film specular reflection of hydrogel lenses (0.04 mm center thickness, 67% nominal water content), which caused inferior arcuate staining. A standard thickness (0.12 mm) lens was worn in the contralateral eye as a control condition. Lenses were worn for a 2 hour period by 20 subjects. Post-lens tear film appearances were categorized as amorphous, faint colored, or colored, where the colored patterns represented a relatively depleted post-lens tear film. We also measured lens dehydration, lens adherence, and pre-lens tear film stability in order to evaluate their role in inferior arcuate staining. The ultrathin and standard lenses caused staining in 100 and 75% of subjects, respectively; the severity of staining was much greater with the ultrathin lenses (Wilcoxon signed ranks test, P = 0.0004). The ultrathin lenses were associated with a higher incidence of post-lens tear film depletion (P = 0.018), had greater front surface dehydration (P = 0.0004), and were more adherent to the eye (paired t-test, P = 0.001). However, pre-lens tear thinning times were not significantly different between the two lens types (Wilcoxon signed ranks test, P > 0.05). These findings support the contention that post-lens tear film depletion is a component of a lens adherence or lens dehydration mediated mechanism of inferior arcuate staining.

Adult↗
PubMed1995

Osmotic determinants of postlens tear film morphology and hydrogel lens movement.

The determinants of postlens tear film (PTF) composition in hydrogel lens wear are poorly understood, although this layer has important roles in lens movement and corneal integrity. We investigated the hypothesis that the PTF could be depleted by instillation of hypotonic saline, using a randomized, double masked, placebo controlled study design. Solutions of 0.90, 0.60 and 0.45% NaCl were instilled into the eyes of 12 subjects wearing ionic and non-ionic high water content hydrogel lenses. Postlens tear film appearances in specular reflection were categorized as amorphous, faint coloured or coloured, where the coloured patterns represent a progressive thinning of the PTF. With instillation of the hypotonic solutions (0.60 and 0.45% NaCl), the appearance of the PTF in specular reflection changed to a faint coloured or coloured pattern in at least 67% of subjects for each lens type (Friedman ANOVA, P < 0.002). For the 0.45% NaCl solution, median lens movement decreased from 0.50 to 0.10 mm (Friedman ANOVA, P = 0.02); however, there were no significant changes in measured lens parameters and no difference between lens types. Postlens tear film depletion due to a hypotonic shift in tear osmolality, as demonstrated here, may explain the clinically observed phenomenon of lens binding.

Adult↗
PubMed1994

Hydrogel (Acuvue) lens movement is influenced by the postlens tear film.

The determinants of hydrogel lens movement with thin mid-water content lenses may be better understood by taking into account the lubrication properties of the tear film. We assessed lens movement, postlens tear film (PTF) status, and tear meniscus height with 58% water content (Acuvue) lenses worn over 6 h on each of 2 consecutive days. Median lens movement was near zero initially and increased within the first hour to 0.34 and 0.30 mm on days 1 and 2, respectively (Friedman analysis of variance by ranks, days 1 and 2, p < 0.001). The small lens movement at 15 min was related to the increased PTF depletion (Kruskal-Wallis analysis of variance; day 1, p = 0.02; day 2, p = 0.007). In some subjects, lens movement and PTF status co-varied throughout the lens wearing periods. These findings suggest the PTF is a determinant of hydrogel lens movement.

Adult↗
PubMed1994

Postlens tear film morphology, lens movement and symptoms in hydrogel lens wearers.

Postlens tear film morphology, lens movement and symptoms were assessed in 100 subjects wearing hydrogel contact lenses on a daily wear schedule. Postlens tear film appearance in specular reflection was categorized as amorphous, coloured, striated or dynamic (variable with blinking), and subjects' symptoms were assessed using a questionnaire. Non-amorphous appearances were present in 25% of subjects and occurred approximately equally with HEMA, Acuvue, Igel 55 and other lens types. Patterned appearances were found to be associated with reduced lens movement (Kruskal-Wallis ANOVA, P < 0.001). The most common symptom (dryness) was not significantly related to postlens tear film appearance. Postlens tear film morphology was shown to be a determinant of lens movement, but may be unrelated to common symptoms.

Adolescent↗
PubMed1993

Isolation and culture of rat superior mesenteric artery smooth muscle cells.

The structure and function of vascular smooth muscle cells have been extensively investigated with the aid of in vitro culture techniques. The majority of studies have utilized aortic tissue as the source of cells. We present here a method for isolating and culturing smooth muscle cells of the rat superior mesenteric artery, an elasto-muscular vessel that is structurally and functionally different from the aorta. Cells were isolated from partially digested explants and characterized by immunochemical and biochemical techniques. Unlike cultured fibroblasts, the cultured cells stained positive for smooth muscle specific actin. The cells also produced laminin and type IV collagen in culture. This method provides a means for the isolation of large numbers of viable smooth muscle cells from the superior mesenteric artery which can be propagated in culture for in vitro study.

Actins↗
PubMed1993

Optimized preparation of Aspergillus fumigatus extracts for allergy diagnosis.

Extracts of Aspergillus fumigatus are required for the measurement of specific antibodies that are important indices in the diagnosis of allergic bronchopulmonary aspergillosis (ABPA). This study investigated the effect of different culture conditions on the production and release of antigenic and allergenic proteins of A. fumigatus. Increasing the incubation temperature from 25 degrees C to 37 degrees C altered the production of proteins by the mycelium which resulted in the release of a greater number of proteins that reacted with precipitating antibodies. Static sporulating cultures produced a much wider antigenic spectrum than shake cultures although the number of precipitating proteins (5 and 3 respectively) and major IgE binding proteins (5 and 3 respectively) was not greatly altered. The widest range of proteins bound by precipitating antibody or IgE from ABPA serum were released into the culture filtrate during 28 day static incubation at 37 degrees C. The resultant extract proved useful for screening patients for specific IgE and will provide a starting material for the identification of individual antigens or allergens.

Allergens↗
PubMed1992

Effects of 4-hydroperoxycyclophosphamide (4-OOH-CP) and 4-hydroperoxydechlorocyclophosphamide (4-OOH-deCICP) on the cell cycle of post implantation rat embryos.

In this study, we used preactivated forms of cyclophosphamide (CP) and dechlorocyclophosphamide (deClCP) to examine the effects of phosphoramide mustard (PM) and acrolein, respectively, on the cell cycle of postimplantation rat embryos. The percentage distribution of cells in the G1/G0, S, and G2/M phases of the cell cycle was determined by flow-cytometric analysis. At embryotoxic concentrations, 4-OOH-CP (PM) induced major cell cycle perturbations whereas 4-OOH-deClCP (acrolein) caused no major perturbation of the cell cycle. These data support the hypothesis that the mechanism of the embryotoxic action of PM involves alkylation of DNA, whereas the mechanism of action of acrolein does not. The primary effect of PM on the cell cycle was an initial delay in the S phase followed by a G2/M arrest. At low embryotoxic concentrations of 4-OOH-CP, there was apparent reversal of the G2/M arrest; at higher embryotoxic concentrations there was little recovery from the G2/M arrest. The high level of cell death found at higher drug concentrations suggests that prolonged G2/M arrest leads to cell death. Using radiolabeled CP and cell sorting, it was determined that PM predominantly alkylated DNA in the S phase of the cell cycle. Overall, the data from this study support the hypothesis that DNA cross-links, induced by the alkylation of DNA by PM, induce cell cycle perturbations. Furthermore, these cell cycle alterations may be one of the early steps in the mechanism leading to the embryotoxicity of PM.

Abnormalities, Drug-Induced↗
PubMed1992

Organization of cells and extracellular matrix in mesenteric arteries of spontaneously hypertensive rats.

Biochemical studies have been used to assess the quantitative changes in elastin and collagen in hypertensive vs. normotensive arteries. However, the relative distribution and organization of these fibrous proteins is likely to be equal in importance to their absolute amounts. In this study we have used scanning electron microscopy in association with selective digestion techniques to assess the organization of cellular and extracellular components of the tunica media of mesenteric arteries of spontaneously hypertensive rats. Superior and small mesenteric arteries were digested with acid, alkali, or bleach to exposure cells, collagen, or collagen and elastin, respectively. We observed that hypertension does not cause a qualitative change in the 3-dimensional arrangement of cells, collagen, or elastin in spontaneously hypertensive arteries when compared to normotensive arteries. However, cells in the superior artery are significantly different in overall shape and surface features when compared to cells of small arteries. These differences in surface morphology of cells are present in hypertensive and normotensive vessels and suggest that superior and small mesenteric artery cells transmit load to the isotropic matrix in different ways. In the elasto-muscular superior artery, force is transmitted across digitations throughout the cell surface. In the muscular small artery, force is transmitted across the tapered, smooth cell surface.

Animals↗
PubMed1992

An investigation of the relationship between microbial and particulate indoor air pollution and the sick building syndrome.

The sick building syndrome has been the subject of research for approximately 10 years. Although it is often suggested that symptoms in office workers are due to circulating micro-organisms or particles, epidemiological studies investigating the relationship between them have been lacking. This cross-sectional study has combined medical and aerobiological assessments of offices in Great Britain and has found that, although airborne particulates and micro-organisms are unlikely to be the sole cause of the sick building syndrome, positive associations between symptom prevalence rates and levels of airborne viable bacteria and fungi within groups of buildings with similar ventilation systems, suggest a possible causal link that should be explored.

Air Microbiology↗
PubMed1992

Three-dimensional characterization of dense bodies in contracted and relaxed mesenteric artery smooth muscle cells.

We have previously shown that dense bodies are not the static planar simple ovoidal structures they appear to be in thin sections. In this report, we present three-dimensional reconstructions from consecutive serial thin sections through shortened and non-shortened large mesenteric artery cells. Profiles of the cell surface, membrane dense bodies, and cytoplasmic dense bodies were reconstructed from consecutive thin sections and the distribution, size, shape, and spatial relationships among these components was examined. Within the cell, membrane dense bodies are numerous and occupy approximately 10% of the cell volume. Membrane dense bodies can attach to the cell surface laterally, obliquely or normally. An individual membrane dense body can be continuous over more than 2 microns of cell depth and can change shape throughout its depth. On cell shortening, many membrane dense bodies assume a crenated shape. Compared to membrane dense bodies, cytoplasmic dense bodies are smaller in all dimensions and occupy about 2% of the cell volume. In shortened cells, cytoplasmic dense bodies appear to cluster into groups. This redistribution of cytoplasmic dense bodies may be related to the reorganization of contractile units when the cell shortens.

Animals↗
PubMed1991

Three-dimensional structure of dense bodies in rabbit renal artery smooth muscle.

In this report, we present a three-dimensional computer assisted reconstruction study from serial thin sections through a rabbit renal artery smooth muscle cell. In a series of 32 consecutive thin (100-nm) sections, one longitudinally oriented cell was followed and photographed in alternating sections. The profiles of the cell surface and dense bodies were reconstructed from these 16 planes and the distribution, size, shape, and spatial relationships among these components was examined. The reconstructed images showed that the cell decreases in diameter from its widest region in the center to the two ends in a step-wise taper. Within the cell, dense bodies are numerous. Relative to the cell axes, a membrane associated dense body (MDB) can be less than or equal to 3.5 microns long, 0.25 micron wide, and may extend up to 2 microns in depth. While the MDB profile in one section may be aligned with the long axis of the cell, in an adjacent section the same dense body may appear almost circular or wedge shaped. The same is true of cytoplasmic dense bodies (CDBs). Compared with MDBs, CDBs are smaller in all dimensions. Some, but not all, CDBs line up in strings oriented with the long axis of the cell. The continuity of dense bodies over considerable cell depth and their change in shape may have important implications for integration of contractile activity and for transmitting passive tension to the extracellular matrix.

Animals↗
PubMed1991

Quantitation and immunohistochemical localization of DNA adducts in rat embryos and associated yolk sac membranes exposed in vitro to N-acetoxy-2-acetylaminofluorene (N-Ac-AAF).

Specific antibodies and radioimmunoassay (RIA) were used to measure the levels of acetylated and deacetylated C-8 substituted deoxyguanosine adducts in day 11 rat embryos and their associated yolk sacs after exposure of whole rat conceptuses in vitro to the teratogen N-acetoxy-2-acetylaminofluorene (N-Ac-AAF). The deacetylated adduct predominates in both the embryo and the associated yolk sac, and a dose response for adduct formation was observed when adducts were quantitated by RIA. Immunohistochemical localization of the deacetylated adducts revealed that adducts were confined to the nuclei in all tissues examined and that the abundance of adducts varied within and between tissues. Our initial findings indicate that specific DNA adduct antibodies may be useful in the study of teratogenesis induced by a wide variety of agents that modify DNA.

Acetoxyacetylaminofluorene↗