Functional and evolutionary characterization of Ohr proteins in eukaryotes reveals many active homologs among pathogenic fungi.

Ohr and OsmC proteins comprise two subfamilies within a large group of proteins that display Cys-based, thiol dependent peroxidase activity. These proteins were previously thought to be restricted to prokaryotes, but we show here, using iterated sequence searches, that Ohr/OsmC homologs are also present in 217 species of eukaryotes with a massive presence in Fungi (186 species). Many of these eukaryotic Ohr proteins possess an N-terminal extension that is predicted to target them to mitochondria. We obtained recombinant proteins for four eukaryotic members of the Ohr/OsmC family and three of them displayed lipoyl peroxidase activity. Further functional and biochemical characterization of the Ohr homologs from the ascomycete fungus Mycosphaerella fijiensis Mf_1 (MfOhr), the causative agent of Black Sigatoka disease in banana plants, was pursued. Similarly to what has been observed for the bacterial proteins, we found that: (i) the peroxidase activity of MfOhr was supported by DTT or dihydrolipoamide (dithiols), but not by ß-mercaptoethanol or GSH (monothiols), even in large excess; (ii) MfOhr displayed preference for organic hydroperoxides (CuOOH and tBOOH) over hydrogen peroxide; (iii) MfOhr presented extraordinary reactivity towards linoleic acid hydroperoxides (k=3.18 (±2.13)×108M-1s-1). Both Cys87 and Cys154 were essential to the peroxidase activity, since single mutants for each Cys residue presented no activity and no formation of intramolecular disulfide bond upon treatment with hydroperoxides. The pKa value of the Cysp residue was determined as 5.7±0.1 by a monobromobimane alkylation method. Therefore, eukaryotic Ohr peroxidases share several biochemical features with prokaryotic orthologues and are preferentially located in mitochondria.

Three-dimensional finite element analysis of stress distribution in peri-implant bone with relined dentures and different heights of healing caps.

The purpose of this study was to evaluate the influence of height of healing caps and the use of soft liner materials on the stress distribution in peri-implant bone during masticatory function in conventional complete dentures during the healing period by using finite element analysis. Three-dimensional models of a severely resorbed mandible with two recently placed implants in the anterior region were created and divided into the following situations: (i) submerged implants, (ii) healing cap at gingival level and (iii) 1·5-mm supragingival. All these situations were also analysed for a conventional complete denture and a denture relined with a 3-mm-thick layer of soft liner material. The models were exported to mechanical simulation software that presented two simulations, one with load in the inferior right canine (35 N) and the other in the inferior right first molar (50 N). Data were evaluated using Maximum Principal Stress provided by the software. All models showed a stress concentration in the cortical bone corresponding to the cervical part of the implant. The simulations with non-submerged implants showed higher values of stress concentration than those that were submerged. Likewise, soft liner materials presented better results than when the denture base was not relined. The height of the healing caps seems to have a direct influence on the stress distribution in the peri-implant bone during the healing period. Considering the values obtained in this study, the use of soft liners with submerged implants seems to be the most suitable method to use during the period of osseointegration.

Effect of exogenous insulin and fasting on growth hormone receptor and IGF-I expression in the pre-ovulatory follicle of ewes.

The aim of this study was to investigate the effect of fasting and exogenous insulin administration on the expression of growth hormone receptor (GHR) and IGF-I mRNA in the pre-ovulatory follicle of ewes. Fifteen ewes received an intravaginal progesterone releasing device that was removed 6 days later (day of removal = day 0). On day -2, the ewes were divided into three groups: (i) fasting group (n = 5) that was fasted from day -2 to day 2; (ii) control group (n = 5) that received a maintenance diet; and (iii) insulin group (n = 5) that received insulin injections (0.25 IU/kg) every 12 h from day -2 to day 2 under the same diet as the control group. Follicular samples were obtained on day 2. Fasting increased plasma non-esterified fatty acids concentrations from day -1 to day 2 (P < 0.001). There was no difference (P > 0.05) in the number of follicles, although there was a tendency for an increase in the pre-ovulatory follicle diameter for the insulin group in comparison to the control group (P = 0.12). Thecal GHR mRNA expression was very low and was considered insignificant. Moreover, granulosa cells GHR mRNA expression increased (P < 0.05) in the insulin group. Expression of IGF-I mRNA was not different among groups in both tissues. In conclusion, insulin administration increases GHR mRNA but not IGF-I mRNA expression in granulosa cells of the pre-ovulatory follicle. However, fasting did not change the pattern of GHR/IGF-I mRNA expression in the pre-ovulatory follicle.

Nipple discharge--study on 100 patients.

In a retrospective study the authors analyzed 100 cases of women with serous, serosanguineous, and sanguineous nipple discharges, without palpable and radiologically visible tumors, that were submitted to resection of main galactophorous ducts and biopsy of underlying breast parenchyma. The sensibility of the cytologic examination of nipple discharges for carcinomas was 16.6%. The histopathology showed incidence of 61% of mammary dysplasia, 33% of intraductal papillomas single or multiple and 6% of carcinomas. The early diagnosis of carcinoma gave good results after radical surgical treatment.

Independent prognostic value of laminin receptor expression in breast cancer survival.

We studied the possible prognostic role of laminin, estrogen (ER), and progesterone (PR) receptors and other pathological factors in relation to the disease-free interval and overall survival of female breast carcinoma patients. Multivariate analyses of clinical and pathological data with respect to the above survival time variables were performed by Cox regression. The statistical dependence of prognosis on ER, PR, and tumor size was based on the discriminant cutoff value that could best distinguish between survival curves. Axillary nodal status was the most significant independent factor in the prediction of both disease-free interval and overall survival of these patients. Use of the information on laminin receptor expression, PR concentration, tumor size, lymphocytic infiltrate, and tumor necrosis improved significantly the prediction of the risk of recurrence. Patients with tumors expressing laminin receptors had 40% less risk of recurrence (P = 0.0209) than those with no expression. On the other hand, four covariates were independently predictive of the risk of death: axillary nodal status, lymphocytic infiltrate, PR and ER concentration. There was a marginally significant (10% level) interaction between tumor size and lymphocytic infiltrate with respect to the prediction of the risk of recurrence. The above sets of variables were used to classify patients into risk groups for the prediction of recurrence and death.