As depicted in Fig. 2 and Fig. 3, dosimetric analysis of PMSF as well as EDTA revealed that at 12 and 50 mM respectively, generation of HA activity was not detectable in serum against hen RBC.

The results presented in Table 6 shows that in both trypsin/α-chymotrypsin-treated serum samples, either of these two RBC types completely adsorbed its own hemagglutinating activity as well as the activity for the other RBC type after 3–4 sequential adsorptions. Pronase, trypsin or SDS-treated sera were found to agglutinate hen RBC within 15 min and was same even up to 120 min (Table 7). HA of trypsin-treated serum against sheep RBC too gave Cilengitide mouse an identical profile (data not shown). The generation of HA activity was detectable in pronase treated serum against hen RBC check details at 25 μg/ml (titer=8), reached highest titer of 256 at 100 μg/ml (Fig. 4), whereas, the HA activity in SDS-treated serum was notable at

0.3 mg/ml (titer=4) and reached the maximum titer at 2 mg/ml (Fig. 5). The untreated serum gave a HA titer of 16 against buffalo and rabbit RBC in the presence of TBS-I or II. In the presence of TBS-IV the serum gave the titers of 8 and 4 against these two RBC types, respectively. Trypsin-treated serum gave a HA titer of 256 against hen RBC, whereas, HA titer against sheep RBC was observed only in the presence of TBS-I and not TBS-II and TBS-IV (Table 8). The pronase-treated serum gave the HA titer of 256 against hen RBC in TBS-II, -III, -IV, -VI, -VII and -VIII (Table 9). Off 53 carbohydrates tested, only eight carbohydrates were found to inhibit HA activity of untreated serum against buffalo RBC at the minimal inhibitory concentrations ranging from 12.5 to 100 mM (Table 10) and these include three glycosides, two disaccharides, two trisaccharides and one tetrasaccharide. Only three hexosamines inhibited the HA activity of pronase treated serum against hen RBC at varying inhibitory concentrations (Table 11). Of the 19 diverse

carbohydrates the HA Avelestat (AZD9668) of trypsin-treated serum against only sheep RBC was inhibited by two N-acetylated aminosugars, at the minimal inhibitory concentration of 25 mM ( Table 12), while, three sialoglycoproteins, inhibited the HA against both hen and sheep RBC with minimal inhibitory concentrations ranging from 0.312 to 2.50 mg/ml ( Table 13). In contrast, asialo-BSM did not inhibit the activity, while, asialo-fetuin did inhibit HA against sheep RBC at a concentration of 1.25 mg/ml and was not inhibitory for agglutination of hen RBC ( Table 13). Human plasma or serum is known to contain two distinct types of naturally occurring humoral molecules capable of causing agglutination of RBC. The first type includes conagglutinins represented by natural antibodies (IgM), whose reactivity is detectable with human RBC and the second type is the lectins naturally occurring in human blood.