Effect of extracorporeal shock wave therapy on pain and function in patients with knee osteoarthritis: a systematic review with meta-analysis and grade recommendations.

OBJECTIVE: To investigate extracorporeal shock wave therapy effect on knee osteoarthritis compared to a sham or kinesiotherapy by a systematic review and meta-analysis of randomized clinical trials. DATA SOURCES: The search was performed in: Cochrane Library, PubMed, PEDro, Web of Science, EMBASE, Scopus, LILACS, and Scielo. REVIEW METHODS: We performed the online search until October, 2022. The following terms were used (Osteoarthritis) AND ("knee joint") AND ("Extracorporeal Shockwave Therapy"). Eligibility criteria: (1) randomized clinical trials; (2) effects comparison of shockwave therapy to a sham or kinesiotherapy in individuals with knee osteoarthritis; (3) pain and physical function as outcome variables. Risk of bias assessed using the PEDro scale. PROSPERO registration (CRD42021235597). RESULTS: We identified 4217 studies, and 12 were included in the qualitative synthesis and the meta-analysis, totaling 403 individuals submitted to the intervention and 331 control individuals. Compared to sham, shockwave was favored in short-term for the function outcome (SMD = -1.93; 95%CI: [-2.77; -1.09]; I² = 83%; P < 0.01). For the pain outcome, the shockwave was favored in the short (MD = -2.05; 95%CI: [-2.59; -1.51]; I² = 84%; P < 0.01), medium (MD = -3.46; 95%CI: [-4.03; -2.89]; I² = 0%; P < 0.01) and long-term (MD = -2.01; 95%CI: [-3.36; -0.65]; I² = 98%; P < 0.01). The association with kinesiotherapy was favored in the short term for the function outcome (SMD = -1.88; 95%CI: [-2.98; -0.78]; I² = 94%; P < 0.01) and favored for the pain outcome in the short (MD = -1.44; 95%CI: [-1.81; -1.07]; I² = 37%; P = 0.14), medium (MD = -1,31; 95%CI: [-1.76; -0,85]; I² = 0%; P = 0.41), and long terms (MD = -1.63; 95%CI: [-1.73; -1.52]; I² = 0%; P = 0.43). CONCLUSION: Shockwave therapy may improve functionality in patients with knee osteoarthritis in the short term and pain in all follow-up moments, compared with sham. When associated to kinesiotherapy, it may improve function in the short term and pain in all follow-up time points, although improvement in pain may not be clinically significant.

Supplementation of sulfate polysaccharides in the seminal cooling medium of common curimata (Prochilodus brevis).

BACKGROUND: The use of sulfated polysaccharides (PS) in seminal cooling is known to improve seminal quality. OBJECTIVE: To evaluate the effect of different concentrations of PS, extracted from the macroalgae Gracilaria domigensis as a supplement to the seminal cooling medium of the reophilic fish Prochilodus brevis (common curimatã). MATERIALS AND METHODS: Five semen pools were diluted in ACP-104 (treatment T1), in BTS® (T2) and in BTS® with different concentrations of PS (0.5 [T3]; 1.0 [T4] and 1.5 [T5]). The samples were cooled for different times (0, 6, 24, 48, 72, 96 and 120 h) and after each hour they were analyzed for: morphology, membrane integrity, DNA integrity and sperm kinetics. RESULTS: There were no significant differences between the treatments containing different concentrations of sulfated polysaccharides. Regarding the different cooling times, it was possible to observe that after hour 96, there was a reduction in the parameters of sperm kinetics. For DNA integrity there was no significant difference in relation to the treatments nor in relation to the hours. For membrane integrity, a reduction was noted as of hour 96, but there was no influence of polysaccharides. For the sperm morphology, there was no statistical difference between the hours, however the BTS was better than the ACP-104. CONCLUSION: It is concluded that the use of polysaccharides in seminal cooling has no negative effect on sperm parameters and proves that seminal cooling keeps the material viable for up to 72 hours. Doi: 10.54680/fr23410110512.

Reduced bone resorption and inflammation in apical periodontitis evoked by dietary supplementation with probiotics in rats.

AIM: To evaluate the relationship between systemic administration of probiotics and inflammation/resorption processes associated with apical periodontitis (AP) in a rat model. METHODOLOGY: Twenty-four male Wistar rats were used. AP was induced in the mandibular left/right first molars. The animals were arranged into three groups: Control, Lactobacillus rhamnosus and L. acidophilus. Probiotics were orally administered via gavage (109 colony-forming units (CFU) diluted in 5 mL of water) for 30 days during the development of AP. On the 30th day, blood was collected to analyse the calcium, phosphorus and alkaline phosphatase concentrations in plasma. Then, the animals were euthanized and the jaws removed for micro-computed tomography and immune-histopathological analysis for receptor activator of NF-κB ligand (RANKL), osteoprotegerin (OPG) and tartrate-resistant acid phosphatase (TRAP). After the Shapiro-Wilk test of normality, the Kruskal-Wallis followed by Dunn's test was performed for nonparametric data, and analysis of variance followed by the Tukey test was performed for parametric data (P < 0.05). RESULTS: There was no significant difference in the calcium and phosphorus levels in plasma amongst the groups (P > 0.05). The level of alkaline phosphatase was significantly higher in the groups that consumed probiotics (P < 0.05). A significantly lower volume of bone resorption was observed in groups that consumed probiotics (P < 0.05). The inflammatory infiltrates and the immunolabelling for RANKL and TRAP were significantly lower in probiotic groups when compared to the control (P < 0.05). Also, the OPG was significantly more immunolabelled in the L. acidophilus group than in the L. rhamnosus and control groups (P < 0.05). CONCLUSION: Probiotic supplementation through gavage (L. rhamnosus and L. acidophilus) had a significant effect on the reduction of inflammation and bone resorption in apical periodontitis development in rats.

Chronic alcohol consumption increases inflammation and osteoclastogenesis in apical periodontitis.

AIM: To investigate the effect of chronic alcohol consumption on apical periodontitis in rats. METHODOLOGY: Thirty-two male Wistar rats were arranged into four groups: Control (C): without apical periodontitis and nonalcoholic diet; (AL): without apical periodontitis and alcoholic diet; (AP): with apical periodontitis and nonalcoholic diet; and (AP + AL): with apical periodontitis and alcoholic diet. The alcoholic solution at 20% was given to the AL and AP + AL groups as the sole source of hydration throughout the experiment. AP was induced in the mandibular left first molars at the end of the 4th week. Weight changes and the amount of solid and liquid foods were recorded for 8 weeks. At the end, the animals were euthanized and the jaws removed followed by histological processing for histopathological and RANKL, OPG, TRAP and HIF-1α analyses. The Mann-Whitney test was used for nonparametric data, and anova followed by the Tukey test was performed for parametric data, with P < 0.05. RESULTS: Animals that received the alcoholic diet had a lower weight gain than the other groups (P < 0.05). Control and AL groups did not have an inflammatory response in the periapical tissues. The median score of inflammatory infiltrate was significantly higher in the AP + AL group (2.5) compared to the AP group (1.5; P < 0.05). In the same comparison, AP + AL was associated with score 3 for RANKL and HIF-1α versus score 2 for AP group (P < 0.05). Moreover, the values for TRAP were 3.88 ± 0.70 cells mm-1 for the AP + AL group and 2.43 ± 0.94 cells mm-1 for the AP group (P < 0.05). CONCLUSION: In rats, an alcoholic diet had a significant effect on the severity of apical periodontitis, exacerbating the inflammatory response and osteoclastogenesis.

Systemic administration of probiotics reduces the severity of apical periodontitis.

AIM: To evaluate the effect of systemic administration of probiotics on the severity of apical periodontitis (AP). METHODOLOGY: Twenty-four male Wistar rats were used. AP was induced in the maxillary left/right first molars. The animals were arranged into groups: Control, Lactobacillus rhamnosus, and Lactobacillus acidophilus. Probiotics were administered orally for gavage (109 colony-forming units diluted in 5 mL of water for 30 days) during the development of AP. After 30 days, cardiac puncture was performed to analyse the complete blood count. Moreover, microbiological analysis of the root canal contents and saliva was performed. Then, the animals were euthanized and the jaw removed for histopathological and IL-10, IL-1ß and IL-6 immunolabeling analyses. After the Shapiro-Wilk test of normality, the Kruskal-Wallis followed by Dunn's test was performed for nonparametric data, and analysis of variance followed by the Tukey test was performed for parametric data (P < 0.05). RESULTS: No significance difference was observed in the blood profiles and in the counts of microorganisms from the saliva samples among the groups (P > 0.05). Total microorganism counts in the root canal, the inflammatory infiltrate and the immunostaining for IL-1ß and IL-6 in AP were significantly lower in the probiotic groups when compared with the control group (P < 0.05). IL-10 was significantly more immunolabled in the probiotic groups than in the control group (P < 0.05). CONCLUSION: Supplementation with probiotics (Lactobacillus rhamnosus and Lactobacillus acidophilus) had a significant effect on the severity of apical periodontitis in rats, demonstrating the anti-inflammatory effect of probiotics on the development of apical periodontitis.

The presence of osteocalcin, osteopontin and reactive oxygen species-positive cells in pulp tissue after dental bleaching.

AIM: To analyse the influence of H2 O2 on pulp repair through osteocalcin and osteopontin immunolabelling and in cellular defence by using the antireactive oxygen species (ROS) antibody. METHODOLOGY: The maxillary molars of 50 rats were treated with 35% H2 O2 (Ble groups) or placebo gel (control groups). At 0 h and 2, 7, 15 and 30 days (n = 10 hemimaxillae), the rats were killed and pulp tissue was evaluated using inflammation and immunolabelling scores (osteocalcin/osteopontin); ROS-positive cells were counted. Paired t-test and Wilcoxon signed-rank test were used (P < 0.05). RESULTS: The Ble group had necrosis in the coronal pulp at 0 h and in the occlusal third of the coronal pulp at 2 days; at 7, 15 and 30 days, no inflammation was noted similar to the controls (P > 0.05). Osteocalcin was absent in the Ble at 0 h, moderate at 2 days and increased thereafter, differing from the controls at all two periods (P < 0.05). Osteopontin was higher principally at 7 and 15 days in Ble groups, but differing with control groups from 2 days after bleaching (P < 0.05). The Ble group had more ROS-positive cells in the pulp at 7 and 15 days (P < 0.05). Tertiary dentine was observed at 7 days, increasing thereafter (P < 0.05). CONCLUSIONS: Post-bleaching pulp repair was associated with increased osteocalcin over time. Osteopontin also participated in this process, and anti-ROS was involved in cellular defence against H2 O2 .

Biocompatibility and immunohistochemical evaluation of a new calcium silicate-based cement, Bio-C Pulpo.

AIM: To evaluate the inflammatory response and ability to induce mineral deposition through histological and immunohistochemical analysis for osteocalcin (OCN), osteopontin (OPN) and bone sialoprotein (BSP) of a new calcium silicate-based cement, Bio-C Pulpo (Angelus), compared to white mineral trioxide aggregate (White MTA-Ang) (Angelus). METHODOLOGY: Polyethylene tubes containing Bio-C Pulpo and White MTA-Ang as well as empty tubes were implanted into the dorsal connective tissue of 30 Wistar rats, which were arranged in five groups according to the period of analysis: 7, 15, 30, 60 and 90 days. After each experimental period, the tubes with surrounding tissue were removed and histologically processed to be analysed using haematoxylin-eosin and immunohistochemistry for the detection of OCN, OPN and BSP. The data were statistically analysed (Friedman's test) at a 5% significance level. RESULTS: The inflammatory response observed with Bio-C Pulpo and White MTA-Ang was greater after 7 and 15 days and decreased from 30 days onwards. No significant difference was found between the control, Bio-C Pulpo and White MTA-Ang at the different periods of analysis (P > 0.05). The immunolabelling for OCN, OPN and BSP was more intense for Bio-C Pulpo and White MTA-Ang after 60 and 90 days, but there was no difference between Bio-C Pulpo and White MTA-Ang at the different periods of analysis (P > 0.05). CONCLUSION: Bio-C Pulpo is biocompatible and induces immunolabelling of osteogenic markers such as OCN, OPN and BSP similar to White MTA-Ang.

RUNX-2, OPN and OCN expression induced by grey and white mineral trioxide aggregate in normal and hypertensive rats.

AIM: To investigate whether hypertension affects mineralization associated with white and grey mineral trioxide aggregate (MTA Angelus® ) implanted subcutaneously into rats by assaying osteoblastic biomarkers. METHODOLOGY: Polyethylene tubes containing grey MTA Angelus® , white MTA Angelus® , intermediate restorative material (IRM; positive control) or an empty tube (negative control) were implanted into the dorsal connective tissue of spontaneous hypertensive (n = 12) and Wistar (normotensive; n = 10) rats. Half of the rats in each group were killed after 7 days, and the remaining after 30 days. Tubes with surrounding tissue were removed, and immunostaining was performed to detect RUNX-2, OPN and OCN proteins. The normality of data was analysed using the Shapiro-Wilk test. Comparison of two independent groups was performed using the Mann-Whitney U-test, to detect a significant difference. A post hoc test accounting for multiple comparisons was performed following Tukey's test (P < 0.05). RESULTS: Under hypertensive conditions after 30 days, both MTA materials were associated with immunolabelling for RUNX-2 from low to moderate, which was less than that observed at normal blood pressure and the 7-day groups (P < 0.05). The expression of OPN and OCN proteins under both MTA conditions was considered low after both 7 and 30 days for the hypertensive condition, and was less than that in animals with normal blood pressure after 30 days (P < 0.05). No immunostaining for any biomarkers in the control and IRM groups was observed (P < 0.05). CONCLUSION: Hypertension decreased the immunostaining of RUNX-2, OPN and OCN biomarkers in response to MTA. Thus, hypertension can jeopardize the mineralization ability of MTA and may have a negative impact on endodontic treatment outcomes.

The effect of dental bleaching on pulpal tissue response in a diabetic animal model: a study of immunoregulatory cytokines.

AIM: To evaluate the influence of tooth bleaching on immunoregulatory cytokines production (IL-6, Tumour necrosis factor (TNF)-α and IL-17) in the pulp tissue of normoglycaemic and diabetic rats. METHODOLOGY: Twenty-eight rats were divided into normoglycaemic and diabetic rats (n = 14). Diabetes mellitus (DM) was induced with a single dose of alloxan diluted in citrate buffer via intramuscular injection. After DM confirmation, all rats were sedated and tooth bleaching was performed using 35% hydrogen peroxide on the right maxillary molars for 30 min. Left molars were used as controls. Bleaching resulted in four hemimaxillae groups: normoglycaemic (N), N-bleached (NBle), diabetic (D) and D-bleached (DBle). After 2 and 30 days, rats were euthanized and hemimaxillae processed for analysis by haematoxylin and eosin and immunohistochemistry. Results within and between animals were submitted to Wilcoxon signed-rank and Mann-Whitney tests (P < 0.05). RESULTS: At 2 days, the NBle group had mild, and the DBle had severe inflammatory infiltration in the pulpal tissue (P < 0.05). TNF-α and IL-6 cytokines were associated with increased immunolabelling in the bleached groups compared to nonbleached (P < 0.05). However, IL-17 had increased immunolabelling in the NBle compared to the N and DBle group (P < 0.05). At 30 days, reactionary dentine was observed in the coronal pulp of all bleached teeth and no inflammation was present (P > 0.05). TNF-α cytokines had increased immunolabelling in the DBle group compared to the D group (P < 0.05). However, for IL-6 and IL-17, no difference was observed in this period (P > 0.05). CONCLUSIONS: Tooth bleaching increased IL-6 and TNF-α in the pulp tissue regardless of diabetes mellitus; however, diabetic rats had higher TNF-α levels for longer periods. Tooth bleaching influenced the increase in IL-17 in the early periods in normoglycaemic rats.

Concentration-dependent effect of bleaching agents on the immunolabelling of interleukin-6, interleukin-17 and CD5-positive cells in the dental pulp.

AIM: To evaluate lymphocyte-like cell activation (CD5-positive cells) and the expression of interleukin (IL)-6 and IL-17 in the pulp after tooth bleaching with two concentrations of hydrogen peroxide (H2 O2 ). METHODOLOGY: The right and left maxillary molars from 40 rats were treated randomly with bleaching gel with 20% H2 O2 (BLUE group, 1 application of 50 min), 35% H2 O2 (MAXX group, three applications of 15 min), or placebo gel (control). After 2 and 30 days, the rats were killed (n = 10), and the jaws were processed for histological and immunohistochemistry analysis of the pulp tissue. The scores of inflammation and immunolabelling (IL-6/IL-17) were submitted to Mann-Whitney and Kruskal-Wallis followed Dunn tests, respectively; anova tests were used for comparisons of number of CD5-positive cells and pulp chamber area values (P < 0.05). RESULTS: At 2 days, 60% of specimens of the BLUE group were associated with moderate inflammation in pulp horns, and in the MAXX group with necrosis (P < 0.05). At 30 days, the pulp was organized, and tertiary dentine was formed. The MAXX group had superior immunolabelling of IL-17 at 2 days differing significantly from other groups (P < 0.05). At 2 days, 90% of the specimens of the BLUE group had moderate immunolabelling of IL-6, and 50% of the MAXX group had severe immunolabelling, both significantly different from the control (P < 0.05). There was no significant difference between the groups at 30 days (P > 0.05). CD5-positive cells were present at 2 and 30 days, particularly in the bleached groups (P < 0.05), without significant difference between time periods (P > 0.05). CONCLUSIONS: IL-6 and IL-17 participated in inflammation in the pulp tissue of rats after tooth bleaching, particularly at 2 days. The immunolabelling was greater with increasing H2 O2 concentration. This process was accompanied by the prolonged activation of CD5-positive cells.

Systemic bone marker expression induced by grey and white mineral trioxide aggregate in normal and diabetic conditions.

AIM: To investigate the relationship between diabetes mellitus and local/systemic effects of both grey and white mineral trioxide aggregate (MTA) Angelus on bone marker expression. METHODOLOGY: Wistar rats were divided into two groups: healthy and diabetic (Alloxan induced), which were further divided into three subgroups (control, GMTA Angelus and WMTA Angelus). Polyethylene tubes filled with MTA materials or empty tubes were implanted in dorsal connective tissue. On days 7 and 30, blood samples were collected for calcium, phosphorus and ALP measurement. The animals were euthanized; implanted tubes were removed and processed for immunohistochemical analysis of osteocalcin (OCN) and osteopontin (OPN). Kruskal-Wallis followed by Dunn's multiple comparison test was performed for nonparametric data, and anova followed by Tukey's test for parametric data. RESULTS: No difference in systemic serum calcium levels between both groups was observed. On day 7, serum phosphorus levels within the WMTA healthy group were higher than that of the diabetic group. On day 30, healthy rats exhibited lower phosphorus levels than diabetic ones. At both time points, the diabetic group was associated with more ALP activity than the healthy group. Immunohistochemical analyses of the healthy group revealed OCN- and OPN-positive cells in the presence of both MTA materials. However, under diabetic conditions, both OCN and OPN were absent. CONCLUSION: Both MTA materials were associated with an increase in serum calcium, phosphorus and ALP, suggesting a potential systemic effect, along with triggered differentiation of OCN- and OPN-positive cells. Moreover, in diabetic conditions, an inhibitory effect on MTA-induced differentiation of OCN- and OPN-positive cells was detected.

Mineral trioxide aggregate improves healing response of periodontal tissue to injury in mice.

BACKGROUND AND OBJECTIVE: Mineral trioxide aggregate (MTA) is a biomaterial used in endodontic procedures as it exerts beneficial effects on regenerative processes. In this study, we evaluate the effect of MTA on healing of periodontal ligament (PDL) and surrounding tissue, following injury, in a transgenic mouse model and on the differentiation of murine mesenchymal progenitor cells in vitro. MATERIAL AND METHODS: We used an inducible Cre-loxP in vivo fate mapping approach to examine the effects of MTA on the contributions of descendants of cells expressing the αSMA-CreERT2 transgene (SMA9+ ) to the PDL and alveolar bone after experimental injury to the root furcation on the maxillary first molars. Col2.3GFP was used as a marker to identify mature osteoblasts, cementoblasts and PDL fibroblasts. The effects of MTA were examined 2, 17 and 30 days after injury and compared histologically with sealing using an adhesive system. The effects of two dilutions of medium conditioned with MTA on proliferation and differentiation of mesenchymal progenitor cells derived from bone marrow (BMSC) and periodontal ligament (PDLC) in vitro were examined using the PrestoBlue viability assay, alkaline phosphatase and Von Kossa staining. The expression of markers of differentiation was assessed using real-time PCR. RESULTS: Histological analyses showed better repair in teeth restored with MTA, as shown by greater expansion of SMA9+ progenitor cells and Col2.3GFP+ osteoblasts compared with control teeth. We also observed a positive effect on differentiation of SMA9+ progenitors into osteoblasts and cementoblasts in the apical region distant from the site of injury. The in vitro data showed that MTA-conditioned medium reduced cell viability and osteogenic differentiation in both PDLC and BMSC, indicated by reduced von Kossa staining and lower expression of osteocalcin and bone sialoprotein. In addition, cultures grown in the presence of MTA had marked decreases in SMA9+ and Col2.3GFP+ areas as compared with osteogenic medium, confirming reduced osteogenesis. CONCLUSION: MTA promotes regeneration of injured PDL and alveolar bone, reflected as contribution of progenitors (SMA9+ cells) into osteoblasts (Col2.3GFP+ cells). In vitro, MTA-conditioned medium fails to promote osteogenic differentiation of both PDLC and BMSC.

The effect of dental bleaching on pulpal tissue response in a diabetic animal model.

AIM: To evaluate pulpal tissue response after dental bleaching in normal and alloxan-induced diabetic rats. METHODOLOGY: Twenty-eight rats were divided into two groups of normoglycaemic and diabetic rats (n = 14). Diabetes mellitus (DM) was induced with alloxan. After DM confirmation, all rats were anaesthetized and dental bleaching was performed with 35% hydrogen peroxide (H2 O2 ) on the right maxillary molars for 30 min. Left molars were used as controls. Bleaching resulted in four hemimaxillae groups: normoglycaemic (N), N-bleached (NBle), diabetic (D) and D-bleached (DBle). After 2 or 30 days, the animals were euthanized and the hemimaxillae were removed, processed for histopathological analysis and stained with haematoxylin-eosin (HE), Masson's trichrome (MT) and picrosirius red (PSR). Results obtained within animals (normoglycaemic or diabetic rats) were submitted to Wilcoxon or paired t-tests, and between animal (normoglycaemic and diabetic rats), to Mann-Whitney test or t-tests. RESULTS: At 2 days, the NBle group had a mild inflammatory infiltration in the pulpal tissue, whilst the DBle had severe inflammation or necrosis (P < 0.05). At 30 days, no inflammation was present. However, a significant difference in pulp chamber area reduction by reactionary dentine deposition was found between the NBle and DBle groups (P < 0.05). At 2 days, fewer immature collagen fibres and more mature collagen fibres were noted in the NBle, D and DBle groups; this was significantly different when compared to the N group (P < 0.05). At 30 days, significantly fewer immature collagen fibres and more mature collagen fibres were noted in NBle compared with DBle group (P < 0.05). CONCLUSIONS: The inflammatory tissue response in rats' teeth after dental bleaching was greater in diabetic rats. Additionally, the increase in reactionary dentine deposition and mature collagen fibres observed in diabetic rats needs further evaluation to confirm the present results.

The contribution of dominance to phenotype prediction in a pine breeding and simulated population.

Pedigrees and dense marker panels have been used to predict the genetic merit of individuals in plant and animal breeding, accounting primarily for the contribution of additive effects. However, nonadditive effects may also affect trait variation in many breeding systems, particularly when specific combining ability is explored. Here we used models with different priors, and including additive-only and additive plus dominance effects, to predict polygenic (height) and oligogenic (fusiform rust resistance) traits in a structured breeding population of loblolly pine (Pinus taeda L.). Models were largely similar in predictive ability, and the inclusion of dominance only improved modestly the predictions for tree height. Next, we simulated a genetically similar population to assess the ability of predicting polygenic and oligogenic traits controlled by different levels of dominance. The simulation showed an overall decrease in the accuracy of total genomic predictions as dominance increases, regardless of the method used for prediction. Thus, dominance effects may not be accounted for as effectively in prediction models compared with traits controlled by additive alleles only. When the ratio of dominance to total phenotypic variance reached 0.2, the additive-dominance prediction models were significantly better than the additive-only models. However, in the prediction of the subsequent progeny population, this accuracy increase was only observed for the oligogenic trait.

Are natural reservoirs important for cholera surveillance? The case of an outbreak in a Brazilian estuary.

UNLABELLED: Paranaguá Bay is one of the largest estuarine systems on the Southern Brazilian coast. The only recorded cholera outbreak in this region since the early 20th century occurred in 1999 and resulted in 467 cases and at least three reported deaths in a population of approx. 150 000 people. This short communication reports historical, unpublished data related to that outbreak. Water, zooplankton and bivalve samples were collected and evaluated using direct fluorescence assay to determine whether Vibrio cholerae serogroups O1 and O139 were present in the estuarine system at that time. Most of the water (83%) and zooplankton samples (75%) were positive for V. cholerae O1, while V. cholerae O139 was not detected. Shellfish (Mytella sp.) were also positive for V. cholerae O1. These results indicate that the estuary, including biological vectors such as copepods and bivalves, comprise an important reservoir of V. cholerae O1 and a probable waterborne pathway for the disease, in addition to contamination with untreated sewage. SIGNIFICANCE AND IMPACT OF THE STUDY: Despite most of the cholera cases that occurred in Brazil during the 7th pandemic were located in the northern areas of the country, a significant outbreak in Paranaguá, an estuary in the south coast, resulted in at least three deaths in 1999. We report here the detection of Vibrio cholerae O1 in water, zooplankton and bivalve samples during the outbreak, using direct fluorescence assay as an alternative method for the traditional plate culture employed at the time by the Brazilian Sanitary Agency. Results demonstrate that aquatic natural reservoirs comprise a potential route of transmission of cholera, in addition to untreated sewage and routine monitoring is recommended.

Identification of sorghum hybrids with high phenotypic stability using GGE biplot methodology.

The aim of this study was to identify sorghum hybrids that have both high yield and phenotypic stability in Brazilian environments. Seven trials were conducted between February and March 2011. The experimental design was a randomized complete block with 25 treatments and three replicates. The treatments consisted of 20 simple pre-commercial hybrids and five witnesses of grain sorghum. Sorghum genotypes were analyzed by the genotype main effects + genotype environment interaction (GGE) biplot method if significant genotype x environment interaction, adaptability, and phenotypic stability were detected. GGE biplot methodology identified two groups of environments, the first composed of Água Comprida-MG, Montividiu-GO, and Vilhena- RO and the second of Guaíra-SP and Sete Lagoas-MG. The BRS 308 and 1G282 genotypes were found to have high grain yield, adaptability, and phenotypic stability and are thus indicated for cultivation in the first and second groups of environments, respectively.

Multi-trait BLUP model indicates sorghum hybrids with genetic potential for agronomic and nutritional traits.

The breeding of sorghum, Sorghum bicolor (L.) Moench, aimed at improving its nutritional quality, is of great interest, since it can be used as a highly nutritive alternative food source and can possibly be cultivated in regions with low rainfall. The objective of the present study was to evaluate the potential and genetic diversity of grain-sorghum hybrids for traits of agronomic and nutritional interest. To this end, the traits grain yield and flowering, and concentrations of protein, potassium, calcium, magnesium, sulfur, iron, manganese, and zinc in the grain were evaluated in 25 grain-sorghum hybrids, comprising 18 experimental hybrids of Embrapa Milho e Sorgo and seven commercial hybrids. The genetic potential was analyzed by a multi-trait best linear unbiased prediction (BLUP) model, and cluster analysis was accomplished by squared Mahalanobis distance using the predicted genotypic values. Hybrids 0306037 and 0306034 stood out in the agronomic evaluation. The hybrids with agronomic prominence, however, did not stand out for the traits related to the nutritional quality of the grain. Three clusters were formed from the dendrogram obtained with the unweighted pair group method with arithmetic mean method. From the results of the genotypic BLUP and the analysis of the dendrogram, hybrids 0577337, 0441347, 0307651, and 0306037 were identified as having the potential to establish a population that can aggregate alleles for all the evaluated traits of interest.

Combining ability of S3 progenies for key agronomic traits in popcorn: comparison of testers in top-crosses.

The successful development of hybrid cultivars depends on the reliability of estimated combining ability of the parent lines. The objectives of this study were to assess the combining ability of partially inbred S3 families of popcorn derived from the open-pollinated variety UENF 14, via top-crosses with four testers, and to compare the testers for their ability to discriminate the S3 progenies. The experiment was conducted in the 2015/2016 crop season, in an incomplete-block (Lattice) design with three replications. The following agronomic traits were evaluated: average plant height, grain yield (GY), popping expansion (PE), and expanded popcorn volume per hectare. The top-cross hybrid, originating from the BRS-Angela vs S3 progeny 10 combination, was indicated as promising, showing high values for specific combining ability for GY and PE. For the S3 progenies that showed high and positive GCA values for GY and PE, the continuity of the breeding program is recommended, with the advance of self-pollination generations. Fasoulas' differentiation index discriminated the BRS-Angela tester as the most suitable for identifying the superior progenies.

Raloxifene modulates regulators of osteoclastogenesis and angiogenesis in an oestrogen deficiency periapical lesion model.

AIM: To analyse the local regulatory mechanisms of osteoclastogenesis and angiogenesis during the progression of periapical lesions in female rats with oestrogen deficiency and treatment with raloxifene (RLX). METHODOLOGY: Female Wistar rats were distributed into groups: SHAM-veh, subjected to sham surgery and treated with a vehicle; OVX-veh, subjected to ovary removal and treated with a vehicle; and OVX-RLX, subjected to ovary removal and treated with RLX. Vehicle or RLX was administered orally for 90 days. During treatment, the dental pulp of mandibular first molars was exposed to the oral environment for induction of periapical lesions, which were analysed after 7 and 30 days. After the experimental periods, blood samples were collected for measurement of oestradiol, calcium, phosphorus and alkaline phosphatase. The rats were euthanized and the mandibles removed and processed for immunohistochemical detection of receptor activator of nuclear factor kappa-B ligand (RANKL), osteoprotegerin (OPG), hypoxia-inducible factor-1 alpha (HIF-1α) and bone-specific alkaline phosphatase (BALP). Data were compared using Kruskal-Wallis followed by Dunn test (nonparametric values) and anova followed by the Tukey's test (parametric values). RESULTS: The plasma concentration of oestradiol showed hypo-oestrogenism in the rats subjected to ovary removal. On day 7, alkaline phosphatase activity, calcium and phosphorus were higher in the OVX-RLX group than in the OVX-veh group (P < 0.001), but immunolabelling for RANKL and HIF-1α was lower in OVX-RLX group (P < 0.001). On day 30, the OVX-veh group had higher immunolabelling for RANKL than the OVX-RLX group (P < 0.05). There were no significant differences in the immunoreactivity of OPG and BALP between any groups at either time-point (P > 0.05). CONCLUSION: RLX therapy reversed the increased levels of the local regulators of both osteoclastogenesis and angiogenesis induced by oestrogen deficiency.

The role of IL-6 on apical periodontitis: a systematic review.

The aim of this review was to examine current knowledge of the role of interleukin-6 (IL-6) in apical periodontitis (AP) pathogenesis as an inflammatory or pro-inflammatory cytokine. It also looked at whether IL-6 could serve as a measure for differential diagnosis or as a biomarker that can further predict the progression of bone resorption. A systematic review relating to AP and IL-6 was made via PubMed, BIOSIS, Cochrane, EMBASE and Web of Science databases using keywords and controlled vocabulary. Two independent reviewers first screened titles and abstracts and then the full texts. The reference lists of the identified publications were examined for additional titles. Eighteen papers were studied in total. In vitro studies (n = 6) revealed that IL-6 is present in AP, and its levels are proportional to the size of the periapical lesions. Neutrophils and macrophages resident in these lesions can produce IL-6 in vitro after a bacterial stimulus. Animal studies (n = 5) showed that IL-6 is present in AP and that osteoblasts can produce IL-6 in vivo. On the other hand, two studies using IL-6 knockout mice revealed larger periapical lesions when compared with control groups, demonstrating IL-6's role as an anti-inflammatory cytokine. In human studies (n = 7), IL-6 was identified in AP, and its levels were higher in symptomatic, epithelialized and large lesions than in asymptomatic and small lesions. These data lead to the conclusion that IL-6 may play a pro-inflammatory role, increasing its levels and reabsorbing bone in the presence of infections. When IL-6 is not present, other cytokines such as IL-1 and TNF-α induce bone resorption. Further studies about the relationship between AP development and the cytokine network must be performed to establish the exact role of each cytokine in the inflammatory process.