In vitro, ex vivo, and in vivo models for dental pulp regeneration.

Based on the concept of tissue engineering (Cells-Scaffold-Bioactive molecules), regenerative endodontics appeared as a new notion for dental endodontic treatment. Its approaches aim to preserve dental pulp vitality (pulp capping) or to regenerate a vascularized pulp-like tissue inside necrotic root canals by cell homing. To improve the methods of tissue engineering for pulp regeneration, numerous studies using in vitro, ex vivo, and in vivo models have been performed. This review explores the evolution of laboratory models used in such studies and classifies them according to different criteria. It starts from the initial two-dimensional in vitro models that allowed characterization of stem cell behavior, through 3D culture matrices combined with dental tissue and finally arrives at the more challenging ex vivo and in vivo models. The travel which follows the elaboration of such models reveals the difficulty in establishing reproducible laboratory models for dental pulp regeneration. The development of well-established protocols and new laboratory ex vivo and in vivo models in the field of pulp regeneration would lead to consistent results, reduction of animal experimentation, and facilitation of the translation to clinical practice.

Genetic diversity and population structure of two Euglossini bee species in a host-parasite relationship.

In the current study, two euglossine species, Exaerete smaragdina and Eulaema nigrita, a cleptoparasite bee and its host, respectively, were used as models to: (i) access the genetic diversity and population structure of both species, sampled along a wide latitudinal range of Atlantic Forest, where the distribution of El. nigrita and Ex. smaragdina co-occurs; (ii) investigate the evolutionary history of these species through the Atlantic Forest, and in a wider scenario, to examine the evolutionary history of these species across others forest domains. Analyses involved males of El. nigrita and Ex. smaragdina sampled through Brazilian territory, including 19 sites in the Atlantic Forest. Bayesian Skyline Plot (BSP) was used to infer possible climate oscillations on population of both species over time. The BSP revealed stability in effective population size for both species in most of the Plio-Pleistocene period. However, BSP results aligned to the starlike configuration in the haplotype network, neutrality test, and population diversity patterns indicated population expansion of the two species during the late Pleistocene. Our findings suggest areas of potential refugia to the climatic oscillations of the Pleistocene in the Atlantic Forest in the Brazilian states of Espírito Santo for El. nigrita and Pernambuco for Ex. smaragdina.

Isolation and characterization of microsatellite markers in two species of the neotropical Epicharis (Hymenoptera, Apidae, Centridini) genus and cross-amplification in related species.

Genetics can provide invaluable tools for management and conservation of bee populations, which are declining worldwide. Among these tools, microsatellite are very useful molecular markers for population analyses. The aim of this study was to isolate and characterize microsatellites for Epicharis (Anepicharis) dejeanii and Epicharis (Epicharis) nigrita, two Neotropical species of solitary bees, both exhibiting the habit of nesting in aggregations. Microsatellite loci were identified from two enriched genomic libraries. The characterization and analysis of loci were carried out using 35 females of E. dejeanii and 34 of E. nigrita. In total, we report the development of 12 microsatellite loci for E. dejeanii and 13 for E. nigrita. For E. dejeanii, all loci were polymorphic, the number of alleles per locus ranged from 2 to 12, averaging 8.7 and, observed and expected heterozygosity were 0.485 (range 0.229-0.857) and 0.633 (range 0.288-0.843), respectively. For E. nigrita, only nine out of 13 loci amplified were polymorphic, the number of alleles per locus ranged from 2 to 12, averaging 5.5. For this species, the observed and expected heterozygosity were 0.440 (range 0.118-0.676) and 0.545 (range 0.167-0.814), respectively. Cross-amplification of primers was successful in other Centridini species. The two sets of loci described for E. dejeanii and E. nigrita species are polymorphic and informative and show promising applicability for both population genetic approaches and relatedness on these and other Centridini species.

Elevated platelet count is a negative predictive and prognostic marker in locally advanced rectal cancer undergoing neoadjuvant chemoradiation: a retrospective multi-institutional study on 965 patients.

BACKGROUND: In patients with locally advanced rectal cancer treated by neoadjuvant chemoradiation, pathological complete response in the surgical specimen is associated with favourable long-term oncologic outcome. Based on this observation, nonoperative management is being explored in the subset of patients with clinical complete response. Whereas, patients with poor response have a high risk of local and distant recurrence, and appear to receive no benefit from standard neoadjuvant chemoradiation. Therefore, in order to develop alternative treatment strategies for non responding patients, predictive and prognostic factors are highly needed. Accumulating clinical observations indicate that elevated platelet count is associated with poor outcome in different type of tumors. In this study we investigated the predictive and prognostic impact of elevated platelet count on pathological response and long-term oncologic outcome in patients with locally advanced rectal cancer undergoing neoadjuvant chemoradiation. METHODS: A total of 965 patients were selected from prospectively maintained databases of seven Centers within the SICO Colorectal Cancer Network. Patients were divided into two groups based on a pre-neoadjuvant chemoradiation platelet count cut-off value of 300 × 109/L identified by receiver operating characteristic curve considering complete pathological response as the outcome. RESULTS: Complete pathological response rate was lower in patients with elevated platelet count (12.8% vs. 22.1%, p = 0.001). Mean follow-up was 50.1 months. Comparing patients with elevated platelet count with patients with not elevated platelet count, 5-year overall survival was 69.5% vs.76.5% (p = 0.016), and 5-year disease free survival was 63.0% vs. 68.9% (p = 0.019). Local recurrence rate was higher in patients with elevated platelet count (11.1% vs. 5.3%, p = 0.001), as higher was the occurrence of distant metastasis (23.9% vs. 16.4%, p = 0.007). At multivariate analysis of potential prognostic factors EPC was independently associated with worse overall survival (HR 1.40, 95% CI 1.06-1.86), and disease free survival (HR 1.37, 95% CI 1.07-1.76). CONCLUSIONS: In locally advanced rectal cancer elevated platelet count before neoadjuvant chemoradiation is a negative predictive and prognostic factor which might help to identify subsets of patients with more aggressive tumors to be proposed for alternative therapeutic strategies.

Food and Alcohol Disturbance in High School Adolescents: Prevalence, Characteristics and Association with Problem Drinking and Eating Disorders.

Food and alcohol disturbance (FAD) is characterized by the association of alcohol use with compensatory behaviors such as restricting calories, physical activity and purging. Despite not being part of the current nosography, research has grown in the past 10 years, mostly on college students' samples. In this study, we aim to describe the prevalence, characteristics and association of FAD with problem drinking (PD) and eating disorder risk (EDR) in a sample of Italian high school students. Participants were 900 high school students (53.6% males; mean age = 16.22) that were administered standardized questionnaires. Students who screened positive for PD, EDR and both were, respectively, 17.3%, 5.9% and 1.3%. Approximately one out four students reported FAD behaviors, mostly to control weight and by restricting calories, with higher prevalence and severity among those who screened positive for PD. Purging behaviors were rare overall (15.5%), but significantly more frequent in participants who screened positive for both PD and EDR (41.7%). FAD was more strongly associated with alcohol use severity than with ED symptom severity across all subgroups. FAD behaviors appear to be common in the Italian high school population and more strongly associated with PD. Future studies should investigate FAD's impact on adolescents' functioning and possible early interventions.

Robotic Rectal Resection for Rectal Cancer in Elderly Patients: A Systematic Review and Meta-Analysis.

Rectal cancer is estimated to increase due to an expanding aging population, thus affecting elderly patients more frequently. The optimal surgical treatment for this type of patient remains controversial because they are often excluded from or underrepresented in trials. This meta-analysis aimed to evaluate the feasibility and the safety of robotic surgery in elderly patients (>70 years old) undergoing curative treatment for rectal cancer. Studies comparing elderly (E) and young (Y) patients submitted to robotic rectal resection were searched on PubMed, Embase, and the Cochrane Library. Data regarding surgical oncologic quality, post-operative, and survival outcomes were extracted. Overall, 322 patients underwent robotic resection (81 in the E group and 241 in the Y group) for rectal cancer. No differences between the two groups were found regarding distal margins and the number of nodes yielded (12.70 in the E group vs. 14.02 in the Y group, p = 0.16). No differences were found in conversion rate, postoperative morbidity, mortality, and length of stay. Survival outcomes were only reported in one study. The results of this study suggest that elderly patients can be submitted to robotic resection for rectal cancer with the same oncologic surgical quality offered to young patients, without increasing postoperative mortality and morbidity.

Biomechanical characterization of a fibrinogen-blood hydrogel for human dental pulp regeneration.

In dental practice, Regenerative Endodontic Treatment (RET) is applied as an alternative to classical endodontic treatments of immature necrotic teeth. This procedure, also known as dental pulp revitalization, relies on the formation of a blood clot inside the root canal leading to the formation of a reparative vascularized tissue similar to dental pulp, which would provide vitality to the affected tooth. Despite the benefit of this technique, it lacks reproducibility due to the fast degradation and poor mechanical properties of blood clots. This work presents a method for constructing a fibrinogen-blood hydrogel that mimics the viscoelastic properties of human dental pulp while preserving the biological properties of blood for application in RET. By varying the blood and fibrinogen concentrations, gels with different biomechanical and biological properties were obtained. Rheology and atomic force microscopy (AFM) were combined to study the viscoelastic properties. AFM was used to evaluate the elasticity of human dental pulp. The degradation and swelling rates were assessed by measuring weight changes. The biomimetic properties of the gels were demonstrated by studying the cell survival and proliferation of dental pulp cells (DPCs) for 14 days. The formation of an extracellular matrix (ECM) was assessed by multiphoton microscopy (MPM). The angiogenic potential was evaluated by an ex vivo aortic ring assay, in which the endothelial cells were observed by histological staining after migration. The results show that the Fbg-blood gel prepared with 9 mg ml-1 fibrinogen and 50% blood of the Fbg solution volume has similar elasticity to human dental pulp and adequate degradation and swelling rates. It also allows cell survival and ECM secretion and enhances endothelial cell migration and formation of neovessel-like structures.

Consequences of the COVID-19 pandemic on the diagnosis and treatment of gastric cancer in referral centers in Italy.

BACKGROUND: The coronavirus pandemic had a major impact in Italy. The Italian health system's re-organization to face the emergency may have led to significant consequences especially in the diagnosis and treatment of malignancies. This study aimed to assess the impact of the pandemic in the diagnosis and treatment of gastric cancer in nine Gruppo Italiano RIcerca Cancro Gastrico (GIRCG) centers. METHODS: All patients assessed for gastric adenocarcinoma at nine GIRCG centers between January 2019 and November 2020 were included. Patients were grouped according to the date of "patient 1's" diagnosis in Italy: preCOVID versus COVID. Clinico-pathological and outcome differences between the two groups were analyzed. RESULTS: A total of 632 patients were included in the analysis (205 in the COVID group). The cT4 weighted ratios were higher in 2020 from April to September, with the greatest differences in May, August and September. The cM+ weighted ratio was significantly higher in July 2020. The mean number of gastrectomies had the greatest reduction in March and May 2020 compared with 2019. The median times from diagnosis to chemotherapy, to complete diagnostic work-up or to operation were longer in 2019. The median time from the end of chemotherapy to surgery was 17 days longer in the preCOVID group. CONCLUSIONS: A greater number of advanced or metastatic cases were diagnosed after the spread of SARS-CoV-2 infection, especially after the "full lockdown" periods. During the pandemic, once gastric cancer patients were referred to one of the centers, a shorter time to complete the diagnostic work-up or to address them to the best treatment option was required.

Aegla buenoi n. sp. (Decapoda: Anomura): first record of aeglid crab from Cinzas River basin, Brazil.

A new species of freshwater anomuran crab, Aegla buenoi n. sp., is described. The new taxon was collected from two streams within the Cinzas River basin, Paran state, Brazil. We used morphological and molecular data (COI mtDNA) to distinguish the new species from its congeners. Aegla buenoi n. sp. is differentiated by morphological diagnostic features of the cephalothorax, chelipeds, second abdominal epimeron, and uropods. Molecular results confirm the separation of A. buenoi n. sp. from closely related species (A. castro Schmitt, 1942, A. lata Bond-Buckup Buckup, 1994, and A. jacutinga Marl Teixeira, 2020). Hence, our study increases the known diversity of aeglids and reports the first species of Aegla from the Cinzas River basin.

Is glyphosate toxic to bees? A meta-analytical review.

Glyphosate (GLY) is an herbicide widely used in agriculture. First considered as non-toxic or slightly toxic to bees, GLY and its different formulations have shown, more recently, to affect negatively the survival, development and behavior of these insects, even when used in doses and concentrations recommended by the manufacturer. Thus, the results of research on the toxicity of GLY to bees are often conflicting, which makes a meta-analysis interesting for data integration, generating a statistically reliable result. Therefore, this study aimed to evaluate the GLY effects on mortality of bees through a meta-analysis. For this, a search was carried out in the databases Web of Science, CAPES (Coordination for the Improvement of Higher Education Personnel - Brazil), Scopus, and PubMed. Papers that evaluated the effect of GLY on bee mortality published between 1945 and October 2020, were considered. After obtaining the data, R software was used to perform the meta-analytical tests. Sixteen papers on mortality were selected with 34 data sets. Most of the sets demonstrated differences between the control and experimental groups, showing that the treatments with GLY caused higher mortality of bees. The results considering the methodology used (ingestion or contact), the phase of the biological cycle (adults or larvae), and the dose (ecologically relevant dose and recommended by the manufacturer) were different when compared with their respective control groups. Therefore, GLY can be considered toxic to bees. It is important to emphasize that this meta-analysis identified that papers assessing the toxicity of GLY to bees are still scarce, for both lethal and sublethal effects, mainly for stingless and solitary bee species.

Mitochondrial DNA diversity of orchid bee Euglossa fimbriata (Hymenoptera: Apidae) populations assessed by PCR-RFLP.

Euglossa fimbriata is a euglossine species widely distributed in Brazil and occurring primarily in Atlantic Forest remnants. In this study, the genetic mitochondrial structure of E. fimbriata from six Atlantic Forest fragments was studied by RFLP analysis of three PCR-amplified mtDNA gene segments (16S, COI-COII, and cyt b). Ten composite haplotypes were identified, six of which were exclusive and represented singleton mitotypes. Low haplotype diversity (0.085-0.289) and nucleotide diversity (0.000-0.002) were detected within samples. AMOVA partitioned 91.13% of the overall genetic variation within samples and 8.87% (phi(st) = 0.089; P < 0.05) among samples. Pairwise comparisons indicated high levels of differentiation among some pairs of samples (phi(st) = 0.161-0.218; P < 0.05). These high levels indicate that these populations of E. fimbriata, despite their highly fragmented landscape, apparently have not suffered loss of genetic variation, suggesting that this particular population is not currently endangered.

Stage IV Gastric Cancer: The Surgical Perspective of the Italian Research Group on Gastric Cancer.

Background/Aim: This work explored the prognostic role of curative versus non-curative surgery, the prognostic value of the various localizations of metastatic disease, and the possibility of identifying patients to be submitted to aggressive therapies. Patients and Methods: Retrospective chart review of stage IV patients operated on in our institutions. Results: Two hundred and eighty-two patients were considered; 73.4% had a single metastatic presentation. In 117 cases, a curative (R0) resection of primary and metastases was possible; 75 received a R1 resection and 90 a palliative R2 gastrectomy. Surgery was integrated with chemotherapy in multiple forms: conversion therapy, HIPEC, neo-adjuvant and adjuvant treatment. Median overall survival (OS) of the entire cohort was 10.9 months, with 14 months for the R0 subgroup. There was no correlation between metastasis site and survival. At multivariate analysis, several variables associated with the lymphatic sphere showed prognostic value, as well as tumor histology and the curativity of the surgical procedure, with a worse prognosis associated with a low number of resected nodes, D1 lymphectomy, pN3, non-intestinal histology, and R+ surgery. Considering the subgroup of R0 patients, the variables pT, pN and D displayed an independent prognostic role with a cumulative effect, showing that patients with no more than 1 risk factor can reach a median survival of 33 months. Conclusions: Our data show that the possibility of effective care also exists for Western patients with stage IV gastric cancer.

Non-anatomical colonic resections: splenic flexure and transverse colectomy. Central vascular ligation is crucial for survival.

BACKGROUND: Cancers of the transverse colon (TC) and of the splenic flexure (SF) are rather uncommon and their prognosis has been reported significantly poorer as compared to right- and left-sided colon cancers. Several studies tried to investigate which is the optimal surgery for oncological and survival outcomes reporting extended colectomies as more effective than limited resections. METHODS: All consecutive patients with diagnosis of colorectal cancer submitted to surgical resection with anastomosis between January 2005 and December 2016 at the Division of Surgical Oncology and Digestive Surgery, Department of Oncology, at the University of Turin, were included. Based on tumor location, patients were enrolled in arm A (cancer of the ascending colon, right (hepatic) flexure, left and sigmoid colon) or arm B (cancer of the transverse colon and of the left (splenic) flexure). RESULTS: Out of 1433 patients registered, 500 patients fulfilled all inclusion and exclusion criteria and were allocated in arm A (N.=425) or in arm B (N.=75). Central vascular ligation and the achievement of at least 5 cm of healthy margin were always performed in all procedures of both arms. Patients' population of the two arms was homogeneous as concerns demographic characteristics and stage of the disease. In arm A, resections were performed more frequently by mini-invasive approach as compared to arm B. Operative time was statistically longer in arm B. Postoperative complications rate was extremely low and comparable in both arms (13.4 and 8.0 in arms A and B, respectively). No postoperative mortality was observed. Overall 5-year survival rates were similar in arm A and B (82.3% and 73.05%, respectively; P=0.29). Arm B patients were treated either with more limited resections (resection of the TC and of the SF) or with extended colectomies (right- and left-sided, standard or enlarged, hemicolectomies), both associated with central vascular ligation. Despite a shorter length of surgical specimen after limited resections, postoperative complications, lymph node yield, and survival were absolutely comparable in both types of surgical treatments. CONCLUSIONS: Limited resections for TC and SF cancers can be performed with the same clinical, oncological and survival outcomes as compared to more extended colectomies if an adequate size of the specimen for the achievement of at least 5 cm of health margin is associated to Central Vascular Ligation.

Transhiatal distal esophagectomy for Siewert type II cardia cancer can be a treatment option in selected patients.

BACKGROUND: While surgical treatment of Siewert I and III (S1,S3) Esophagogastric Junction (EGJ) cancer is codified, the efficay of transhiatal procedure with anastomosis in the lower mediastinum for Siewert II (S2) still remains a dibated topic. METHODS: This is a large multicenter retrospective study. The results of 598 consecutive patients submitted to resection with curative intent from January 2000 to January 2017 were reported. Clinical and oncological outcomes of different procedures performed in S2 tumor were analyzed to investigate the efficacy of transhiatal approach. RESULTS: The 5-year overall survival rate (OS) was poor (32%) for all Siewert types. The most performed operations in S2 cancer were proximal gastrectomy + transthoracic esophagectomy (TTE or Ivor-Lewis procedure, 60%), total gastrectomy + transhiatal distal esophagectomy with anastomosis in the chest (THE, 24%) and total gastrectomy + transthoracic esophagectomy (TGTTE, 15%). Cardiovascular and pulmonary complications were higher after TTE. On the contrary, surgical complications were significantly higher after THE. Postoperative mortality was similar. The distribution of TNM stages was different in the 3 types of procedures: patients submitted to THE had an earlier stage disease. With this bias, OS after THE was higher than after TTE but the difference was not significant (49.85% vs 28.42%, p = 0.0587). CONCLUSIONS: Despite a higher rate of postoperative surgical complications, OS after total gastrectomy and transhiatal distal esophagectomy was at least comparable to that of transthoracic approach in less advanced S2 tumors. Therefore, THE with anastomosis in the chest could be a treatmen option in earlier S2 tumors.

Comparative analysis of adaptive and neutral markers of Drosophila mediopunctata populations dispersed among forest fragments.

Comparison of adaptive and neutral genetic markers is a valuable approach to characterize the evolutionary consequences of populations living in environments threatened by anthropogenic disturbances, such as forest fragmentation. Shifts in allele frequencies, low genetic variability, and a small effective population size can be considered clear signs of forest fragmentation effects (due to genetic drift) over natural populations, while adaptive responses correlate with environmental variables. Brazilian Atlantic Forest had its landscape drastically reduced and fragmented. Now, several forest remnants are isolated from each other by urban and crop areas. We sampled Drosophila mediopunctata populations from eight forest remnants dispersed on two adjacent geomorphological regions, which are physiognomic and climatically quite distinct. Microsatellite data of inversion-free chromosomes (neutral genetic marker) indicate low structuration among populations suggesting that they were panmictic and greatly influenced by gene flow. Moreover, significant differences in chromosomal inversion frequencies (adaptive genetic marker) among populations and their correlations with climatic and geographical variables indicate that genetic divergence among populations could be an adaptive response to their environment. Nonetheless, we observed a significant difference in inversion frequencies of a population in two consecutive years that may be associated with edge and demographic effects. Also, it may be reflecting seasonal changes of inversion frequencies influenced by great temperature variation due to edge effects. Moreover, the forest fragment size does not affect genetic variation of neutral markers. Our data indicate that despite oscillations in chromosomal inversion frequencies, D. mediopunctata populations from Brazilian Atlantic Forest and their divergence may be driven by adaptive factors to local differences, perhaps because it is a small flying insect easily carried by the wind increasing its migration rates.

Neoadjuvant chemotherapy for gastric cancer. Is it a must or a fake?

AIM: To investigate the neoadjuvant chemotherapy (NAC) effect on the survival of patients with proper stomach cancer submitted to D2 gastrectomy. METHODS: We proceeded to a review of the literature with PubMed, Embase, ASCO and ESMO meeting abstracts as well as computerized use of the Cochrane Library for randomized controlled trials (RCTs) comparing NAC followed by surgery (NAC + S) with surgery alone (SA) for gastric cancer (GC). The primary outcome was the overall survival rate. Secondary outcomes were the site of the primary tumor, extension of node dissection according to Japanese Gastric Cancer Association (JGCA) performed in both arms, disease-specific (DSS) and disease-free survival (DFS) rates, clinical and pathological response rates and resectability rates after perioperative treatment. RESULTS: We identified a total of 16 randomized controlled trials comparing NAC + S (n = 1089) with SA (n = 973) published in the period from January 1993 - March 2017. Only 6 of these studies were well-designed, structured trials in which the type of lymph node (LN) dissection performed or at least suggested in the trial protocol was reported. Two out of three of the RCTs with D2 lymphadenectomy performed in almost all cases failed to show survival benefit in the NAC arm. In the third RCT, the survival rate was not even reported, and the primary end points were the clinical outcomes of surgery with and without NAC. In the remaining three RCTs, D2 lymph node dissection was performed in less than 50% of cases or only recommended in the "Study Treatment" protocol without any description in the results of the procedure really perfomed. In one of the two studies, the benefit of NAC was evident only for esophagogastric junction (EGJ) cancers. In the second study, there was no overall survival benefit of NAC. In the last trial, which documented a survival benefit for the NAC arm, the chemotherapy effect was mostly evident for EGJ cancer, and more than one-fourth of patients did not have a proper stomach cancer. Additionally, several patients did not receive resectional surgery. Furthermore, the survival rates of international reference centers that provide adequate surgery for homogeneous stomach cancer patients' populations are even higher than the survival rates reported after NAC followed by incomplete surgery. CONCLUSION: NAC for GC has been rapidly introduced in international western guidelines without an evidence-based medicine-related demonstration of its efficacy for a homogeneous population of patients with only stomach tumors submitted to adequate surgery following JGCA guidelines with extended (D2) LN dissection. Additional larger sample-size multicentre RCTs comparing the newer NAC regimens including molecular therapies followed by adequate extended surgery with surgery alone are needed.

Surgical treatment of hepatic metastases from gastric cancer.

The purpose of the study was to investigate the clinical factors influencing the prognosis of patients submitted to hepatectomy for metastases from gastric cancer and their clinical role. We conducted a retrospective multicentre review. We evaluated how survival from surgery was influenced by patient-related, tumour-related and treatment-related prognostic factors. We analysed data on 144 patients submitted to hepatectomy for metastases from gastric cancer, in the synchronous and metachronous setting. In 117 cases, an R0 resection was achieved, while in 27 an R + hepatic resection was performed. Chemotherapy was administered to 55 patients. Surgical mortality was 2.1% and morbidity 21.5%. One-, 3-, and 5-year OS rates after surgery were 49.9, 19.4 and 11.6%, respectively, with a median OS of 12.0 months. T4 gastric cancer, H3 hepatic involvement, non-curative resection, recurrence after surgery, and abstention from chemotherapy were associated with a worse prognosis. Factor T and H displayed a clear (p < 0.001) cumulative effect. Our data show that R0 resection must be pursued whenever possible. The treatment of T4 gastric cancer with hepatic bilateral and diffuse metastasis (H3) should be considered carefully or it should be probably avoided. Finally, a multimodal treatment associating surgery and chemotherapy offers the best survival results.

Expression of cyp1a induced by benzo(A)pyrene and related biochemical and genotoxic biomarkers in the neotropical freshwater fish Prochilodus lineatus.

The goal of this work was to design specific cyp1a primers for the fish Prochilodus lineatus to study the expression of this gene and its relation to the activity of biotransformation phase I enzyme (ethoxyresorufin O-deethylase - EROD) and genotoxic damage after 6 and 24 h of benzo(a)pyrene (B(a)P) intraperitoneal injection. In comparison to fish injected only with canola oil (vehicle), the expression of cyp1a and EROD activity both in the liver and gills were significantly higher after 6 and 24 h of B(a)P injection. A significant increase in DNA damage was detected in liver and blood cells after 6 h of B(a)P injection and in the gill cells after both times, probably caused by intermediate metabolites of B(a)P. Thus, the expression of cyp1a and its relationship with the corresponding enzyme activity is a potential biomarker for evaluation P. lineatus exposure to organic compounds.

A new species of Aegla Leach, 1820 (Crustacea, Anomura) from the Iguaçu River basin, Brazil.

A new species of aeglid crab, Aegla okora n. sp., is described. This species was found in the Iguaçu River basin in Paraná state, southern Brazil. Aegla okora n. sp. was discriminated based on morphological and molecular data (mitochondrial COI). Morphologically, the new species differs from congeners of the same river basin by the presence of a trapezoidal areola and the ventromesial border of the ischium having three tubercles, besides other exclusive additional morphological characteristics. Molecular results demonstrated that there is no overlap between the intraspecific distances of Aegla okora n. sp. and the interspecific distance of other species of Aegla added to the analysis, confirming the separation of species and increasing the known diversity of the Iguaçu River basin.