Pleiotropic roles of LAMMER kinase, Lkh1 in stress responses and virulence of Cryptococcus neoformans.

Dual-specificity LAMMER kinases are highly evolutionarily conserved in eukaryotes and play pivotal roles in diverse physiological processes, such as growth, differentiation, and stress responses. Although the functions of LAMMER kinase in fungal pathogens in pathogenicity and stress responses have been characterized, its role in Cryptococcus neoformans, a human fungal pathogen and a model yeast of basidiomycetes, remains elusive. In this study, we identified a LKH1 homologous gene and constructed a strain with a deleted LKH1 and a complemented strain. Similar to other fungi, the lkh1Δ mutant showed intrinsic growth defects. We observed that C. neoformans Lkh1 was involved in diverse stress responses, including oxidative stress and cell wall stress. Particularly, Lkh1 regulates DNA damage responses in Rad53-dependent and -independent manners. Furthermore, the absence of LKH1 reduced basidiospore formation. Our observations indicate that Lkh1 becomes hyperphosphorylated upon treatment with rapamycin, a TOR protein inhibitor. Notably, LKH1 deletion led to defects in melanin synthesis and capsule formation. Furthermore, we found that the deletion of LKH1 led to the avirulence of C. neoformans in a systemic cryptococcosis murine model. Taken together, Lkh1 is required for the stress response, sexual differentiation, and virulence of C. neoformans.

Continued development of the root separated from the main root.

INTRODUCTION: Endodontic infection or physical trauma of a developing tooth might lead to unusual situation for root development. METHODS: Two cases are presented. Both cases involved immature mandibular second premolars with infected pulps. In the first case, the tooth was treated by apexification, whereas revascularization procedures were performed in the second case. Interestingly, a separated root tip was observed with the main root at the initial presentation of case 2. RESULTS: At recall after apexification, a separate root tip was observed apically to the main root end, and root development continued in case 1. In the second case, the separate root displayed continuing root formation, but the main root did not gain root thickness or length. CONCLUSIONS: These cases revealed that Hertwig's epithelial root sheath and stem cells from the apical papilla can be separated from the main tooth structure by an external force or iatrogenic factors and thereafter produce a separate root tip.

An evaluation of buccal infiltrations and inferior alveolar nerve blocks in pulpal anesthesia for mandibular first molars.

We compared the anesthetic efficacy of inferior alveolar nerve blocks (IANBs) with that of buccal infiltrations (BIs) in mandibular first molars. Using a crossover design, all subjects received a standard IANB or a BI of 1.7 mL of 4% articaine with 1:100,000 adrenaline (Septanest; Septodont, Saint-Maru-des-Fosses, France) on two appointments separated by at least 1 week. Pulpal anesthesia was determined by using an electric pulp tester. Electric pulp testing was repeated at 5, 8, 11, 15, 20, 25, and 30 minutes after the injections. Anesthesia was considered successful if the subject did not respond to the maximum output of the pulp tester at two or more consecutive time points. Fifty-four percent of the BI and 43% of the IANB were successful; the difference was not significant (p = 0.34). The onset of pulpal anesthesia was significantly faster with BI (p = 0.03). In conclusion, BI with 4% articaine for mandibular first molars can be a useful alternative for clinicians because compared with IANB it has a faster onset and a similar success rate.

Effect of smear layer and chlorhexidine treatment on the adhesion of Enterococcus faecalis to bovine dentin.

The aim of this in vitro study was to determine the effects of a smear layer and chlorhexidine (CHX) treatment on the adhesion of Enterococcus faecalis to bovine dentin. Forty dentin blocks from bovine incisors were prepared and randomly divided into four groups of 10 each. The blocks in group 1 were placed in sterile saline for 5 minutes, while those in group 2 were treated with 17% EDTA for 5 minutes. The blocks in group 3 were placed in 2% CHX for 7 days. The blocks in group 4 were treated with 17% EDTA for 5 minutes, and then placed in 2% CHX for 7 days. All the blocks were immersed in a suspension of E. faecalis for 3 hours. The bacteria adhering to the dentin surface were counted by examination using a scanning electron microscope. The most significant amount of bacteria was retained on the samples from group 1 (p < 0.05) and the smallest amount of bacteria adhered to the samples from group 4. These results suggest that a smear layer enhances the adherence of E. faecalis to the dentin, and CHX is effective in reducing the adherence of microorganisms.

Effect of different temperatures and penetration depths of a System B plugger in the filling of artificially created oval canals.

OBJECTIVES: The purpose of this study was to compare the gutta-percha-filled area in oval canals while varying the temperature and penetration depth of the System B plugger. STUDY DESIGN: Seventy-five resin blocks, with artificially created oval canals, were obturated by using a continuous wave technique. In groups 1, 2, and 3, the plugger was introduced to a depth of 4 mm from the apex at set temperatures of 100, 200, and 350 degrees C, respectively. In group 4, the plugger was inserted to a depth of 2 mm from the apex at a set temperature of 200 degrees C. Group 5 was obturated in the same way as group 2, and then an additional cold plugging was performed to a depth of 3 mm from the apex. The cross sections of the obturated canals were photographed, and the percentage of the gutta-percha-filled area (PGP) was measured. RESULTS: No significant differences were found between the 3 different temperature settings (P >.05). At 1 mm from the apex, group 4 showed the highest PGP (95.57% +/- 6.86), followed by group 5 (84.72% +/- 12.19), and group 2 (67.33 +/- 13.03) (P <.05). CONCLUSION: In artificially created oval canals, plugger temperature within the range used in this study did not affect the PGP. However, the penetration depth of the plugger was significantly related to PGP.