Antigenicity of influenza virus hemagglutinin following chemical modification

Hemagglutinin (HA) molecules from a number of different strains of type A influenza virus were reacted with 1-fluoro 2–4-dinitrobenzene (FDNB) at pH values ranging from 8.4 to 10.3. HA was also reacted with tetranitromethane (TNM) or diazotized sulfanilic acid (DSA). Sequence studies on HA1 from HA molecules treated with FDNB, TNM, or DSA showed that certain lysine, tyrosine, or histidine residues were 100% substituted after the reaction, while others apparently did not react at all. DNP-substituted hemagglutinin molecules, isolated from FDNB-treated A/Memphis/1/71_H-BEL_N(H3N1) virus particles, had up to 58% of lysines substituted with DNP. These molecules, nevertheless, retained hemagglutinin activity and, as far as could be measured, the same capacity as the unsubstituted hemagglutinin to react with heterogeneous antiserum or a panel of monoclonal antibodies. These results suggest that those amino acid side chains able to react with FDNB (lysine, histidine, tyrosine, and cysteine) are either not present in the antigenic sites on the HA, or if they are, then either the side chains which bind antibody do not react with DNP, or the presence of DNP in the site does not affect its ability to combine with antibody. The results also suggest that substitution of more than half of the lysine in the hemagglutinin molecule does not cause any marked conformational changes, for such changes would be expected to affect the ability of the HA to combine with both cell receptors and antibody molecules. Similar findings with TNM-treated HA suggest that tyrosine is not an essential part of any antigenic site on H3 type HA. HA treated with diazotized sulfanilic acid lost HA activity, but its antigenicity was similar to that of untreated HA when tested with heterogeneous antisera, suggesting that histidine was not present in the antigenic sites. However, when HA from a monoclonal variant of A/Mem/1/71 (H3N2) virus with a sequence change from wild-type in HA1 of proline (143) to histidine (Laver, Air, and Webster, 1981) was reacted with diazotized sulfanilic acid, the histidine at position 143 in HA1 reacted completely and the HA lost the ability to bind antibody specific for the new antigenic site on this variant. However, treatment of this variant with FDNB did not lead to substitution of histidine 143.     

Declining density of intimal smooth muscle cells and age as preconditions for atheronecrosis in the basilar artery

The aging basilar artery has some differences and some similarities when compared with the aorta and coronary arteries. As the non-necrotic intimal thickness increases over time, the number of smooth muscle cells reaches a steady state around age 25–30 years in the coronaries and aorta, but continues to increase in the basilar artery, even to 90 years of age. The numbers of cells per unit of tissue (the cell density) declines with age, and the patterns of decline are quantitatively similar in all three arterial segments. All arteries so far examined behave alike in showing that atheronecrosis emerges in those specimens that have sufficiently low density of intimal smooth muscle cells. These results identify low intimal cell density as a criterion for recognizing arteries that are prone to atheronecrosis. One possible explanation is that depopulation of the fibrotically thickened and aged intima, by spreading apart the smooth muscle cells with expanding matrix materials, could be the conditioning factor that brings about the intrusion of atheronecrosis.     

Direct and indirect effects of soluble extracts of Schistosoma mansoni eggs on fibroblast proliferation in vitro.

The possibility that soluble products of Schistosoma mansoni eggs might participate in the pathogenesis of hepatic fibrosis in schistosomiasis was investigated. Both crude saline extracts of eggs (soluble egg antigen [SEA]) and a partially purified SEA fraction contained activity which stimulated guinea pig and human dermal fibroblasts to proliferate in vitro, as measured by uptake of [3H]thymidine. Maximum activity was present in fractions which eluted from Sephacryl S-200 with an apparent molecular weight of less than or equal to 12,500 and in fractions which had an estimated pI 8, as determined by preparative isoelectric focusing of partially purified SEA. Activity in crude SEA was not removed by chromatography on concanavalin A-Sepharose 4B. When concanavalin A-binding glycoproteins lacking intrinsic fibroblast-stimulating activity were incubated with spleen cells from infected or uninfected mice, fibroblasts-stimulating activity was detected in the culture supernatants. Thus, SEA contains two functionally distinct molecular species. One of these directly stimulates fibroblasts, whereas the other induces the release of a fibroblast-stimulating activity from lymphocytes or macrophages or both. Since these fibroblast-stimulating factors might be elaborated in the livers of infected individuals, these observations suggest a potential role of soluble schistome products in the pathogenesis of hepatic fibrosis in schistosomiasis.     

Poly(lactide-co-glycolide) microspheres as a moldable scaffold for cartilage tissue engineering

This study demonstrates the use of biodegradable poly(lactide-co-glycolide) (PLG) microspheres as a moldable scaffold for cartilage tissue engineering. Chondrocytes were delivered to a cylindrical mold with or without PLG microspheres and cultured in vitro for up to 8 weeks. Cartilagenous tissue formed using chondrocytes and microspheres maintained thickness, shape, and chondrocyte collagen type II phenotype, as indicated by type II collagen staining. The presence of microspheres further enhanced total tissue mass and the amount of glycosaminoglycan that accumulated. Evaluation of microsphere composition demonstrated effects of polymer molecular weight, end group chemistry, and buffer inclusion on tissue-engineered cartilage growth. Higher molecular weight PLG resulted in a larger mass of cartilage-like tissue formed and a higher content of proteoglycans. Cartilage-like tissue formed using microspheres made from low molecular weight and free carboxylic acid end groups did not display increases in tissue mass, yet a modest increased proteoglycan accumulation was detected. Microspheres comprised of PLG with methyl ester end groups yielded a steady increase in tissue mass, with no real increase in matrix accumulation. The microencapsulation of Mg(OH)(2) had negative effects on tissue mass and matrix accumulation. The data herein reflect the potential utility of a moldable PLG-chondrocyte system for tissue-engineering applications.     

肺神经内分泌癌的免疫组化与免疫电镜研究

本文应用免疫组化与免疫电镜技术对20例肺神经内分泌癌进行了研究,另6例非神经内分泌癌作为对照。结果表明:嗜铬颗粒蛋白A免疫组化染色19/20例阳性,免疫电镜能够比较特异地标记神经分泌颗粒,具有确诊意义;神经特异性烯醇化酶染色20/20例阳性,非神经内分泌癌3/6例阳性,免疫电镜下无特异性分布,适宜作为诊断的筛选指标;S-100蛋白和5-HT亦可作为诊断之参考。     

Effects of exercise on vasodilatory capacity in endurance- and resistance-trained men

To determine vasodilatory responsiveness we measured forearm blood flow (FBF) following reactive hyperemia (RH), prior to and following a bout of maximal aerobic exercise in endurance- (n=14) and resistance-trained men (n=10). Both groups were similar in height, body mass, and percentage body fat. Using strain-gauge plethysmography, resting FBF was higher in the resistance-trained group [4.82 (0.84) vs 3.33 (1.17) ml min⁻¹ 100 ml⁻¹ of tissue; P<0.05]. However, the resistance-trained group had a 17%–29% lower pre-exercise FBF response to RH for the first 45 s (P<0.05). Following the maximal exercise bout there were no group differences in FBF. Post-exercise FBF was higher compared to pre-exercise values in both the endurance- (P<0.001) and resistance- (P<0.01) trained groups. Endurance-trained men appear to have a greater peak vasodilatory capacity compared to resistance-trained men, and acute maximal exercise increased the vasodilatory capacity in both groups. Acute exercise also equalized the peak vasodilatory response between the endurance- and resistance-trained groups, suggesting the potential for flow-mediated vasodilatation was similar for both groups. Electronic Publication     

A modified human ELISPOT assay to detect specific responses to primary tumor cell targets

Background  

The desired outcome of cancer vaccination is to induce a potent T cell response which can specifically recognize and eliminate autologous tumor cells in vivo. Accordingly, immunological assays that demonstrate recognition of native tumor cells (tumor-specific) may be more clinically relevant than assays that demonstrate recognition of tumor protein or peptide (antigen-specific).     

Cell adhesion-related proteins as specific markers of sponge cell types involved in allogeneic recognition

Sponge immunocyte identification is of interest to comparative immunologists since characterizing these cells will allow investigations into the mechanisms of non-self recognition in the oldest animal phylum. Here, we report that polyclonal antibodies raised against the core protein of a proteoglycan involved in cell adhesion in the marine sponge Microciona prolifera are specific markers for archaeocytes, the totipotent sponge cells. Archaeocytes are mobilized upon allogeneic contact and they accumulate in the contact zone. A second type of cell, the gray cells, are specifically recognized by monoclonal antibodies raised against CD44, a hyaluronan receptor. Gray cells do also accumulate in the contact area. Specific staining of a third sponge cell type, the rhabdiferous cells, shows that these do not accumulate upon allografting. These specific cell markers allow tracking of archaeocytes and gray cells, and show that they play an active role in sponge allogeneic reactions.     

Genetic diagnoses and associated anomalies in fetuses prenatally diagnosed with esophageal atresia

Esophageal atresia (EA) is a congenital anomaly occurring in 2.3 per 10,000 live births. Due to advances in prenatal imaging, EA is more readily diagnosed, but data on the associated genetic diagnoses, other anomalies, and postnatal outcome for fetuses diagnosed prenatally with EA are scarce. We collected data from two academic medical centers (n = 61). Our data included fetuses with suspected EA on prenatal imaging that was confirmed postnatally and had at least one genetic test. In our cohort of 61 cases, 29 (49%) were born prematurely and 19% of those born alive died in the first 9 years of life. The most commonly associated birth defects were cardiac anomalies (67%) and spine anomalies (50%). A diagnosis was made in 61% of the cases; the most common diagnoses were vertebral defects, anal atresia, cardiac anomalies, tracheoesophageal fistula with esophageal atresia, radial or renal dysplasia, and limb anomalies association (43%, although 12% met only 2 of the criteria), trisomy 21 (5%), and CHARGE syndrome (5%). Our findings suggest that most fetuses with prenatally diagnosed EA have one or more additional major anomaly that warrants a more comprehensive clinical genetics evaluation. Fetuses diagnosed prenatally appear to represent a cohort with a worse outcome.