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Behavioural and electrophysiological responses ofAnopheles gambiaeGilessensustricto (Diptera: Culicidae) to human skin emanations collected on glass beads were studied using a dual-port olfactometer and electroantannography. Glass beads to which skin emanations from human hands had been transferred elicited a level of attraction similar to a human hand. The attractiveness of these handled glass beads faded away 4 h after transfer onto the beads. Storage at−20°C for up to 8 weeks showed a decreased but still attractive effect of the beads. In a choice test between one individual and four others, the emanations from the reference individual were significantly more attractive in three out of four cases. The headspace of handled glass beads elicited a dose-dependent EAG response. The substances causing EAG activity could be removed partially by dichloromethane, ethanol and pentane-ether. Glass beads provide a suitable neutral substrate for the transfer of human odour to enable chemical analysis of the human skin emanations for identification of kairomones of anthropophilic mosquitoes. [ABSTRACT FROM AUTHOR]

Objective: To seek the optimal melanin-removal method for hematoxylin and eosin (HE) staining, immunohistochemistry and molecular detection. Methods: Thirty-eight paraffin tissue samples of malignant melanoma diagnosed at the Fujian Cancer Hospital, Fuzhou, China between January 2018 and March 2022 were collected and used to make a tissue microarray. Melanin in these cases was removed using warm hydrogen peroxide, double oxidation depigmentation, modified potassium permanganate-oxalic acid or trichloroisocyanuric acid, followed by HE staining. The cases were divided into two cohorts: one was subject to the one of the above four methods to remove melanin first, followed by immunohistochemistry (SOX-10, Ki-67, HMB45 and Melan A), while the other was subject to immunohistochemical staining first and then a melanin removal. Following that, seventeen melanin-rich paraffin tissue samples were collected and depigmented using the methods described above. DNA extraction was then done, followed by assessments of DNA content and quality. Moreover, the completeness of melanin removal, the effect on HE and immunohistochemical staining, and the quality of DNA were compared between the depigmented methods. Results: Regarding the effectiveness of melanin removal, the modified potassium permanganate-oxalic acid and the warm hydrogen peroxide methods were the most effective, and both showed residual melanin in only 5.26% (2/38) of the cases. The trichloroisocyanuric acid method showed residual melanin in 10.53% (4/38) of the cases. The worst was the double oxidation depigmentation method, which showed pigment residue in 15.79% (6/38) of the cases. For HE staining, the percentage of good staining with the warm hydrogen peroxide method was 92.11%, higher than the other three methods. For immunohistochemical staining, the mean staining scores of immunohistochemistry first followed by melanin removal with modified potassium permanganate-oxalic acid, double oxidation and trichloroisocyanuric acid were 20.84, 26.63 and 35.02, respectively. These immunohistochemical staining scores were higher than those of melanin removal first followed by immunohistochemistry (8.70, 15.41 and 21.22, respectively). The mean staining score of melanin removal by warm hydrogen peroxide method followed by immunohistochemistry was 33.57, superior to that of immunohistochemistry followed by the melanin removal (19.96). Moreover, the staining scores of HMB45, MelanA and Ki-67 with immunohistochemical staining followed by trichloroisocyanuric acid method were 36.45, 33.79, and 36.24, respectively, while the staining score of SOX10 with melanin removal by warm hydrogen peroxide followed by immunohistochemistry was 34.39. The DNA was significantly degraded by modified potassium permanganate-oxalic acid, double oxidation depigmentation and trichloroisocyanuric acid, whereas the mean concentration of DNA extracted after melanin removal by hydrogen peroxide method was 59.59 μg/L, substantially higher than that of DNA extracted without melanin removal (30.3 μg/L, P =0.001). The A 260 / A 280 of DNA extracted after melanin removal by hydrogen peroxide was between 1.8 and 2.0 in all cases, and the A 260 / A 230 was above 2.0 in sixteen cases, suggesting high purity of DNA. However, the DNA extracted without removing the melanin showed poor purity, with A 260 / A 280 below 1.8 in eight cases and A 260 / A 230 below 2.0 in sixteen cases. Conclusions: Warm hydrogen peroxide showed the least melanin residue, superior HE staining and a minimal effect on DNA purity/quality compared to the other three methods. It thus appears most suitable for PCR, NGS and other molecular detection. Melanin removal with trichloroisocyanuric acid after immunohistochemical staining has the least melanin residual, and thus could be the most convenient and efficient. However, it is noted that the efficacy of the same depigmentation method varies with different antibodies. Therefore, the optimal depigmentation method should be selected based on the specific markers of interest.

Aim: To compare the kinetics of positive inotropism between ibopamine (Ibo) and ouabain (Oua)., Methods: The isolated right papillary muscle of cat was used to assess the positive inotropic effects by cummulative concentrations of Ibo and Oua. The maximal effects was recognized as the contraction just before the appearance of spontaneous contraction. A Hill equation was developed by using lg [E/(Emax-E)] vs lg C., Results: The S value of Ibo was smaller than that of Oua, causing C95/C5 (ratio of concentrations producing 95%/5% of Emax) of Ibo (264) elevenfold as large as Oua (22.9). Comparing with Oua, concentrations of Ibo initiating effects were lower, the slope was smaller, and higher concentrations were required for producing maximal effect. During drug concentration declining, the effect-concentration curve of Ibo showed a counter-hysteresis loop, more remarkable than Oua., Conclusion: The changes in positive inotropism of Ibo against drug concentrations were less than those of Oua.

The indole-derived compounds, which possessed side chains resembling those of changrolin (4-[3',5'-bis[(N-pyrrolidinyl)methyl]-4'-hydroxyaniline]-quinazoline) showed potent anti-arrhythmic activity by restoration of sinus rhythm from ouabain-induced tachycardia in guinea pigs. The potency was assessed by comparison of the maintenance time of sinus rhythm recovered from tachyarrhythmias induced by ouabain. The promising compound was MI2 with piperidyl residue on position 3 & 5 of phenol moiety. There was no difference in anti-arrhythmic activities resulting from substitutions between a benzene ring and methyl residue at position 2 of indole, but the latter had weaker parasympatholytic activity. The anti-arrhythmic activity of MI2 (greater than 60 min) was 2.4 times more potent then changrolin (25 min), but its anti-cholinergic activity was only half of the latter. To compare the suppressive effect on reperfusion-induced arrhythmias by iv MI2 at different time in relation to the ligation-reperfusion protocol, it was the most effective when administered either 30 min prior to coronary occlusion or at the moment of reperfusion. The compound MI might belong to the Ic group shown by the slowing impulse conduction within the heart.