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Table 1.

Origin of insects used and the estimated protein content per one salivary gland.

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Figure 1.

Hyaluronidase activity of insects tested by dot method on polyacrylamide gel with incorporated hyaluronan.

Protein content per 2 µl dot is indicated in brackets. Tb = Tris buffer, Sh = sheep testicular hyaluronidase (10 µg), Rp = Rhodnius prolixus SGE (20 µg), Ph = Pediculus humanus BE (20 µg), Cf = Ctenocephalides felis BE (20 µg), Pp = Phlebotomus papatasi SGE (0.8 µg), Ck = Culicoides kibunensis BE (20 µg), As = Anopheles stephensi SGE (0.8 µg), Cq = Culex quinquefasciatus SGE (0.8 µg), Aa = Aedes aegypti SGE (1.3 µg), Oo = Odagmia ornata SGE (0.8 µg), El = Eusimulium latipes SGE (1.7 µg), Cv = Chrysops viduatus SGE (3.5 µg), Gf = Glossina fuscipes SGE (14 µg), Sc = Stomoxys calcitrans SGE (2.4 µg).

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Figure 2.

Substrate specificity of hyaluronidases tested on polyacrylamide gel with incorporated chondroitin sulfate.

Protein content per 2 µl dot is indicated in brackets. Sh = sheep testicular hyaluronidase (10 µg), Cf = Ctenocephalides felis SGE (20 µg), Cq = Culex quinquefasciatus SGE (0.8 µg), Ck = Culicoides kibunensis BE (20 µg), Pp = Phlebotomus papatasi SGE (0.8 µg), Cv = Chrysops viduatus SGE (0.8 µg).

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Figure 3.

SDS PAGE zymography under nonreducing conditions on polyacrylamide gel with incorporated hyaluronan.

Protein content per lane is given in brackets. Pp = Phlebotomus papatasi SGE (0.2 µg), Ck = Culicoides kibunensis BE (10 µg), Cq = Culex quinquefasciatus SGE (5.6 µg), El = Eusimulium latipes SGE (0.4 µg), Oo = Odagmia ornata SGE (0.4 µg), Cv = Chrysops viduatus SGE (0.2 µg), Cf = Ctenocephalides felis BE (15 µg).

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Figure 4.

SDS PAGE zymography on the same gel as in Figure 3 but under reducing conditions.

Pp = Phlebotomus papatasi SGE (0.2 µg), Ck = Culicoides kibunensis BE (20 µg), Cq = Culex quinquefasciatus SGE (8 µg), El = Eusimulium latipes SGE (3 µg), Oo = Odagmia ornata SGE (3 µg), Cv = Chrysops viduatus SGE (0.3 µg), Cf = Ctenocephalides felis BE (15 µg).

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Figure 5.

Effect of SDS on hyaluronidase activity in SGE of Culex quinquefasciatus.

Activity of hyaluronidase was tested by dot method on polyacrylamide gel with incorporated 0.002% hyaluronan and 0.001% SDS. Protein content per 2 µl dot is indicated in brackets. Sh = sheep testicular hyaluronidase (2 µg), Pp = Phlebotomus papatasi SGE (0.8 µg), Cq1 = Culex quinquefasciatus SGE (1.6 µg), Cq2 = Culex quinquefasciatus SGE (3.2 µg).

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Figure 5 Expand

Figure 6.

Effect of hyaluronidase on Leishmania infection in mice.

BALB/c mice were coinoculated intradermally into ear with 104 or 105 Leishmania major and hyaluronidase equivalent to 0, 2 and 10 salivary glands of Phlebotomus papatasi. Lesion size, given as a product of its area (mm2) and the degree of ulceration (1–5), was monitored for 6 weeks post infection. Points (▪) = mean values, boxes = 95% confidence intervals, whiskers = min-max values. The p values of corresponding Kruskal-Wallis ANOVA are provided.

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Figure 6 Expand