Structural studies of human purine nucleoside phosphorylase: Towards a new specific empirical scoring function

Human purine nucleoside phosphorylase (HsPNP) is a target for inhibitor development aiming at T-cell immune response modulation. In this work, we report the development of a new set of empirical scoring functions and its application to evaluate binding affinities and docking results. To test these new functions, we solved the structure of HsPNP and 2-mercapto-4(3H)-quinazolinone (HsPNP:MQU) binary complex at 2.7 A resolution using synchrotron radiation, and used these functions to predict ligand position obtained in Clocking Simulations. We also employed molecular dynamics simulations to analyze HsPNP in two conditions, as apoenzyme and in the binary complex form, in order to assess the structural features responsible for stability. Analysis of the Structural differences between systems provides explanation for inhibitor binding. The use of these scoring functions to evaluate binding affinities and molecular docking results may be used to guide future efforts on virtual screening focused on HsPNP. (c) 2008 Elsevier Inc. All rights reserved.4791283

Evaluation of Different Passive Ultrasonic Irrigation Protocols on the Removal of Dentinal Debris from Artificial Grooves

Abstract The aim of this study was to evaluate the influence of different passive ultrasonic irrigation protocols on the removal of debris in artificially created grooves in the cervical, middle and apical root thirds. Forty extracted bovine incisor roots were instrumented to 1 mm of the root apex with a R50 Reciproc instrument and irrigated with 2.5% sodium hypochlorite. The roots were then inserted in a muffle and cleaved into two hemisections. Grooves (3-mm long) were done at 2, 7 and 12 mm from the root apex in one hemisection and filled with dentinal debris. The hemisections were regrouped into the muffle and divided into 4 groups (n=10) according to the final irrigation protocol: Control group: 3x20 s using a #30 gauge needle without agitation of the irrigating solution; Group PUI-s (static passive ultrasonic irrigation): 3x20 s of passive ultrasonic irrigation with the tip of the insert maintained static on the apical third; Group PUI-t (passive ultrasonic irrigation per third): 20 s of PUI in each third; Group PUI-d (passive dynamic ultrasonic irrigation): 3x20 s of PUI dynamically moving the insert in the whole extent of the root canal. In all groups, was used a total of 6 mL of 2.5% sodium hypochlorite as irrigant. After these procedures, the grooves were analyzed with a stereomicroscope and assigned scores as regards removal of the debris. The data were analyzed statistically (a=0.05). The results showed a better cleaning in all the groups where the irrigating solution was agitated with ultrasonic device than in the control group (p<0.05). In the apical third, the PUI-d and PUI-s showed similar performance (p>0.05) and a better cleanness than PUI-t (p<0.05). The dynamic and static methods of agitation of the irrigating solution provided more effective cleaning. PUI-d provided the most completely clean grooves suggesting that its use is the most adequate in cases of teeth with complex canal anatomy

Effect of Passive Ultrasonic Irrigation on Enterococcus faecalis from Root Canals: An Ex Vivo Study

Endodontic irrigation aims to clean and disinfect the root canal system. Passive ultrasonic irrigation (PUI) is based on the use of an ultrasound-activated instrument into the root canal filled with irrigant. The aim of this study was to evaluate, ex vivo, the effectiveness of PUI in eliminating Enterococcus faecalis from root canals. Seventy-five extracted human single-root teeth were used. After root canal preparation, specimens were inoculated with E. faecalis and incubated at 37 °C for 21 days. Specimens were distributed into five groups (n=15), according to the irrigation method: PUI + saline solution (PUI/SS); PUI + 1% NaOCl (PUI/NaOCl); conventional needle irrigation (CNI) + saline solution (CNI/SS); CNI + 1% NaOCl (CNI/NaOCl); No irrigation (control). Microbiological samples were collected at three time points: initial (21 days after inoculation), post-irrigation (immediately after irrigation), and final (7 days after irrigation). Data were obtained in CFU mL-1 and subjected to analysis by ANOVA and Tukey’s tests at 5% significance level. The post-irrigation samples did not demonstrate statistical difference between PUI/SS and CNI/SS nor between PUI/NaOCl and CNI/NaOCl (p>0.05), but PUI/NaOCl and CNI/NaOCl had lower CFU mL-1 number than the other groups (p>0.05). Statistically significant difference was observed between the initial and post-irrigation samples and between the post-irrigation and final samples (p<0.05) in all groups, except in the control. The final samples of all groups presented bacterial counts similar to the initial samples. PUI or CNI with 1% NaOCl contribute to disinfection, but are unable to eradicate E. faecalis from the root canal system

Interactions Between Bt-Bioinsecticides and Podisus nigrispinus (Dallas) (Hemiptera: Pentatomidae), a Predator of Plutella xylostella (L.) (Lepidoptera: Plutellidae)

Bioinsecticides are being increasingly used to protect vegetable crops against herbivores, but data on the side effects of such strategy on the third trophic level are still required. We investigated the influence of the Bacillus thuringiensis subsp. kurstaki HD-1 strain and of the commercial bioinsecticide Agree® on the biological aspects of the predator Podisus nigrispinus (Dallas) (Hemiptera: Pentatomidae) when feeding on Plutella xylostella (L.) (Lepidoptera: Plutellidae)-infected larvae. On average, infected larvae were consumed more often than the non-infected larvae throughout the predator nymphal development, and no effect on nymph survival was observed. Population growth parameters (R0, rm, and λ) resulting from the fertility life tables did not differ among treatments. The results led to the conclusion that P. nigrispinus can be used in combination with the product Agree® or the strain HD-1 for the control of P. xylostella. The combined use of these control agents helps to maintain the balance of the ecosystem and contributes for the production of food for healthy human consumption.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Lab de Biologia e Criação de Insetos, Depto de Fitossanidade, FCAV/UnespDepto de Biologia, Univ de São PauloLab de Biologia e Criação de Insetos, Depto de Fitossanidade, FCAV/Unes