Ultrastructural Aspects of The Intestinal Cells Surface of Angiostrongylus Cantonensis By Using of Transmission Electron Microscopy.

The rat lungworm Angiostrongylus cantonensis is a zoonotic parasite and the main cause of eosinophilic meningitis in the world. Its main source of nutrients originates from the degradation of host hemoglobin in blood-feeding helminths, such as A. cantonensis. The purpose of this study was to analyze the ultrastructure of the intestinal cells by using of transmission electron microscopy (TEM). A. cantonensis worms obtained from Rattus norvegicus (norway rats) from endemic area were used for TEM. The ultrastructural analysis was performed using fragments cut from the middle part of the worms, and the TEM study revealed the cells with microvilli and nuclei containing areas of loose and condensed chromatin and the presence of macrovesicles and microvesicles of digestion and it was observed in this study that intestinal epithelium is look like a syncytium. The ultrastructural aspects of the intestinal cells surface of A. Cantonensis the indicate that the intestinal epithelium is a multinucleate mosaic or syncytium.

Poor survival after resection of early gastric cancer: extremes of survivorship analysis reveal distinct genomic profile.

A subset of patients with early gastric cancer demonstrate early recurrence and poor survival despite margin-negative resection. This study used an extremes-of-survivorship approach to identify an association between TP53 hotspot mutations co-occurring with loss of heterozygosity and unexpectedly poor survival in early gastric cancer. This distinct genomic profile may be a novel biomarker of poor survival in patients with completely resected early gastric cancer, and warrants large-scale validation. Promising, validation needed.

Effectiveness and safety of Omalizumab in the treatment of chronic spontaneous urticaria: Systematic review and meta-analysis.

INTRODUCTION: Chronic spontaneous urticaria (CSU) affects approximately 1% of the population, affecting both children and adults. Omalizumab (Oma) is a therapeutic option for patients with refractory forms of CSU. OBJECTIVES: To determine the effectiveness and safety of Oma in the treatment of CSU. METHODS: Systematic review (Cochrane Collaboration methodology) of randomized clinical trials comparing Oma to placebo in refractory CSU treatment. The search is based on MEDLINE; EMBASE, Central Cochrane Library, and LILACS. The outcomes evaluated were: control of the illness, adverse events, and quality of life. RESULTS: Of the 848 identified studies 13 were selected for further review and six were included in the meta-analysis. For all outcomes, high-quality evidence has confirmed that Oma is effective in the treatment of CSU. The dosage of 300mg/month achieved better results; namely a significant reduction in pruritus, papules, and urticaria activity, as well as an increase in the number of patients with a controlled condition, improvement in the quality of life and no differences in adverse events compared to the placebo. CONCLUSIONS: High-quality evidence demonstrates that Oma is effective and safe in the treatment of CSU refractory to therapy with H1 antihistamines.

Low-level laser therapy and anesthetic infiltration for orofacial pain in patients with fibromyalgia: a randomized clinical trial.

BACKGROUND: To compare the analgesic effect of anesthetic infiltration of lidocaine 2% and low-level laser therapy (LLLT) by GaAlAs into tender points of patients with orofacial pain and fibromyalgia (FM). MATERIAL AND METHODS: A randomized clinical trial was performed with adults (N=66) that were allocated into two groups (1:1): Group A received LLLT irradiation by Diode Laser GaAlAs (780nm) with expositions twice a week during six weeks and Group B was treated with anesthetic infiltration of lidocaine 2% without vasoconstrictor once a week for four weeks. The pain assessment included the Visual Analogic Scale (VAS) and tenderness to palpation. RESULTS: No dropout and adverse effect was observed during the study. The pain decreased significantly in each group after the treatment (p=0.0001, ß=1.0), even though no statistical difference was found between both treatments (p=0.46, ß= 0.82). The presence of tender points decreased after both treatments, with responsively in some types of masticatory muscles (p<0.05) except posterior temporalis muscle. The patients perception showed that both treatments were effective and a few patients reported that the treatment did not improve welfare. CONCLUSIONS: The LLLT by GaAlAs and anesthetic infiltration of lidocaine 2% were equally effective to control orofacial pain in FM individuals.

Effect of feeding strategies and cropping systems on greenhouse gas emission from Wisconsin certified organic dairy farms.

Organic agriculture continues to expand in the United States, both in total hectares and market share. However, management practices used by dairy organic producers, and their resulting environmental impacts, vary across farms. This study used a partial life cycle assessment approach to estimate the effect of different feeding strategies and associated crop production on greenhouse gas emissions (GHG) from Wisconsin certified organic dairy farms. Field and livestock-driven emissions were calculated using 2 data sets. One was a 20-yr data set from the Wisconsin Integrated Cropping System Trial documenting management inputs, crop and pasture yields, and soil characteristics, used to estimate field-level emissions from land associated with feed production (row crop and pasture), including N2O and soil carbon sequestration. The other was a data set summarizing organic farm management in Wisconsin, which was used to estimate replacement heifer emission (CO2 equivalents), enteric methane (CH4), and manure management (N2O and CH4). Three combinations of corn grain (CG) and soybean (SB) as concentrate (all corn = 100% CG; baseline = 75% CG + 25% SB; half corn = 50% CG + 50% SB) were assigned to each of 4 representative management strategies as determined by survey data. Overall, GHG emissions associated with crop production was 1,297 ± 136 kg of CO2 equivalents/t of ECM without accounting for soil carbon changes (ΔSC), and GHG emission with ΔSC was 1,457 ± 111 kg of CO2 equivalents/t of ECM, with greater reliance on pasture resulting in less ΔSC. Higher levels of milk production were a major driver associated with reduction in GHG emission per metric tonne of ECM. Emissions per metric tonne of ECM increased with increasing proportion of SB in the ration; however, including SB in the crop rotation decreased N2O emission per metric tonne of ECM from cropland due to lower applications of organically approved N fertility inputs. More SB at the expense of CG in the ration reduced enteric CH4 emission per metric tonne of ECM (because of greater dietary fat content) but increased N2O emission per metric tonne of ECM from manure (because of greater N content). An increased reliance on pasture for feed at the expense of grain resulted in decreased in milk production, subsequently leading to substantially higher emissions per metric tonne of ECM.

Hemocytes of Rhipicephalus sanguineus (Acari: Ixodidae): Characterization, Population Abundance, and Ultrastructural Changes Following Challenge with Leishmania infantum.

Few studies have examined the cellular immune response of ticks, and further research on the characterization of the hemocytes of ticks is required, particularly on those of Rhipicephalus sanguineus (Latreille) because of the medical and veterinary importance of this tick. The aims of this study were to characterize the morphology and the ultrastructure of the different types of hemocytes of adult R. sanguineus and to determine the population abundance and the ultrastructural changes in the hemocytes of ticks infected with Leishmania infantum. The hemocytes were characterized through light and transmission electron microscopy. Within the variability of circulating cells in the hemolymph of adult R. sanguineus, five cell types were identified, which were the prohemocytes, plasmatocytes, granulocytes, spherulocytes, and adipohemocytes. The prohemocytes were the smallest cells found in the hemolymph. The plasmatocytes had polymorphic morphology with vesicles and cytoplasmic projections. The granulocytes had an elliptical shape with the cytoplasm filled with granules of different sizes and electrodensities. The spherulocytes were characterized by several spherules of uniform shapes and sizes that filled the entire cytoplasm, whereas the adipohemocytes had an irregular shape with multiple lipid inclusions that occupied almost the entire cytoplasmic space. The total counts of the hemocyte population increased in the group that was infected with L. infantum. Among the different cell types, the numbers increased and the ultrastructural changes occurred in the granulocytes and the plasmatocytes in the infected group of ticks.

Preoperative somatostatin analogues versus direct transsphenoidal surgery for newly-diagnosed acromegaly patients: a systematic review and meta-analysis using the GRADE system.

UNLABELLED: Whether the preoperative use of somatostatin analogues (SA) improves surgical outcomes in acromegaly is still a matter of debate. OBJECTIVE: We conducted a systematic review of randomized, controlled trials that compared the short-term outcomes of preoperative use of SA (Pre-SA) with direct TSS (No-SA) for the treatment of newly diagnosed acromegaly. METHODS: Embase, Pubmed, Lilacs, and Central Cochrane were used as our data sources. The primary outcomes were no need for any adjuvant treatment 3 months after surgery, based on biochemical results (GH nadir after OGTT <1 µg/L and normal IGF-1 for age and gender), quality of life and mortality. The included trials were analyzed using the Grading of Recommendations Assessment, Development, and Evaluation approach. RESULTS: A total of 2.099 references were identified and two reviewers independently screened the titles and abstracts. From the 14 potentially eligible studies, four were included and ten were excluded due to lack of randomization or different outcomes. A pool of 261 patients was randomly assigned to Pre-SA or No-SA. Meta-analysis of IGF1 normalization showed a significant difference in favor of Pre-SA (RR 2.47; 95% CI 1.66, 3.77). Adding a GH nadir on OGTT ≤1 µg/L, we found a RR of 2.15 (95% CI 1.39, 3.33). Quality of evidence for no need of adjuvant postoperative treatment was moderate, but for improving quality of life was very low and for mortality was absent. CONCLUSION: Pre-SA increases the chance of biochemical control of acromegaly 3 months after TSS in patients harboring GH-secreting pituitary macroadenomas.

Techniques for restoring enlarged canals: an evaluation of fracture resistance and bond strength.

AIM: To verify the influence of fibreglass post diameter, as well as use of accessory posts on fracture resistance and bond strength, when used for restoring enlarged root canals. METHODOLOGY: One hundred maxillary single-rooted canine human teeth were decoronated and root canals were prepared using a No.4 drill (White Post, FGM, Joinville, SC, Brazil). The roots were assigned to five groups: (EC1) post No.1 (Exacto Cônico, Angelus, Londrina, PR, Brazil); (EC2) post No.2 (Exacto Cônico, Angelus); (EC3) post No.3 (Exacto Cônico, Angelus); (ECA) post No.1 (Exacto Cônico, Angelus) plus two accessory posts (Reforpin, Angelus); (WP) post No.4 (White Post, FGM). Posts 1-4 have a crescent diameter. Posts were luted (Rely X-Arc, 3M ESPE, USA) and composite resin filling cores were prepared for the fracture resistance test (n = 10). For the push-out test (n = 10), roots were sectioned into 1 mm thick slices. Both tests were performed in a universal testing machine. Data were analysed using anova and Tukey's test. RESULTS: Groups WP and EC3 had higher fracture resistance than the other groups (P < 0.05), which were statistically similar. Root fractures occurred in 14% of the specimens. Groups EC1 and EC2 had lower bond strength values than Groups EC3 and ECA which were lower than the WP Group (P < 0.05). Bond strength was lower in the apical and middle third than in the cervical third of root canals (P < 0.05). CONCLUSIONS: Thicker posts were associated with higher resistance to fracture and bond strength to root canals. Using a post plus two accessory posts improved the bond strength but not the fracture resistance.

Manufacturing ceramic bricks with polyaluminum chloride (PAC) sludge from a water treatment plant.

The objective of this research work is to assess the viability of manufacturing ceramic bricks with sludge from a water treatment plant (WTP) for use in real-world applications. Sludge was collected from settling tanks at the Bolonha WTP, which is located in Belém, capital of the state of Pará, Brazil. After dewatering in drainage beds, sludge was added to the clay at a local brickworks at different mass percentages (7.6, 9.0, 11.7, 13.9 and 23.5%). Laboratory tests were performed on the bricks to assess their resistance to compression, water absorption, dimensions and visual aspects. Percentages of 7.6, 9.0, 11.7 and 13.9% (w/w) of WTP sludge presented good results in terms of resistance, which indicates that technically, ceramic bricks can be produced by incorporating up to 13.9% of WTP sludge.

Comparison of peripartum metabolic status and postpartum health of Holstein and Montbéliarde-sired crossbred dairy cows.

Objectives of the current experiment were to evaluate plasma concentrations of metabolites and haptoglobin peripartum, uterine health and involution, and follicle growth and resumption of cyclicity of Holstein (HO) and Montbéliarde-sired crossbred cows. Cows (52 HO and 52 crossbred) were enrolled in the study 45 d before expected calving date. Cows had body weight and body condition score recorded on d -45, -14, 0, 1, 28, and 56 relative to calving. Dry matter intake was calculated for a subgroup of cows (25 HO and 38 crossbred) from 6 wk before to 6 wk after calving. Blood was sampled weekly from d -14 to 56 relative to calving for determination of glucose, nonesterified fatty acid, and ß-hydroxybutyrate concentrations; from d -7 to 21 relative to calving for determination of haptoglobin concentration; and from d 14 to 56 postpartum for determination of progesterone concentration. Cows were examined at calving and on d 4, 7, 10, and 14 postpartum for diagnosis of postparturient diseases, on d 24 postpartum for diagnosis of purulent vaginal discharge, and on d 42 postpartum for diagnosis of subclinical endometritis. Uteri and ovaries were examined by ultrasonography every 3 d from d 14 to 41 postpartum. Milk yield and composition were measured monthly and yield of milk, fat, protein, and energy-corrected milk were recorded for the first 90 d postpartum. Body weight was not different between Holstein and crossbred cows, but HO cows had reduced body condition score compared with crossbred cows. Even though DMI from 6 wk before to 6 wk after calving tended to be greater for HO cows (16.8 ± 0.7 vs. 15.3 ± 0.5 kg/d), HO cows tended to have more pronounced decline in dry matter intake, expressed in percentage of body weight from d -15 to 0 relative to calving. Energy-corrected milk and nonesterified fatty acid and ß-hydroxybutyrate concentrations were not different between breeds. No differences were observed in incidence of retained fetal membranes, metritis, and subclinical endometritis, but HO cows tended to be more likely to have pyrexia from d 0 to 15 postpartum (50.0 vs. 31.4%) and to have greater incidence of purulent vaginal discharge (44.2 vs. 26.5%) than crossbred cows. Holstein cows were more likely to have at least 1 uterine disorder postpartum than crossbred cows (63.5 vs. 36.7%). No differences between breeds were observed in uterine involution. Holstein cows had larger subordinate follicles (10.1 ± 0.4 vs. 8.9 ± 0.5) and a greater number of class III follicles (1.6 ± 0.1 vs. 1.2 ± 0.1) than crossbred cows. Furthermore, the first corpus luteum postpartum of HO cows was diagnosed at a slower rate compared with crossbred cows. Crossbred cows had improved uterine health compared with HO cows and this may have been a consequence of heterosis and (or) breed complementarity and less pronounced decrease in DMI during the last days of gestation.

Diet and between-tissue isotope comparisons reveal different foraging strategies for age and sex of a Saffron Finch (Sicalis flaveola Linnaeus, 1766) population.

Measuring stable isotopes in different tissues offers the opportunity to provide insight into the foraging ecology of a species. This study aimed to assess how diet varies between yellow females, yellow males, and dull individuals of a Saffron Finch (Sicalis flaveola) population. We measured δ13C and δ15N in blood over a year, and in different feathers, to estimate seasonal consistency of resource use for each category. We conducted this study in a private farm in the Central Brazilian savannas. We sampled 195 individuals in seven field samplings between January 2017 and March 2018. The mean blood δ13C values were similar among yellow females, yellow males and dull individuals, indicating that this population of Saffron Finch predominantly accesses similar resources throughout the year, with a predominant C4 signal. Although Saffron Finch is considered a granivorous species, the mean δ15N values found indicate that both adults and juveniles also incorporate in their tissues some invertebrate. The slight isotope-tissue difference between feathers and blood is similar to the reported in previous studies and may reflect tissue-to-tissue discrimination. The isotopic space of yellow males was greater than that of yellow females and dull individuals, indicating greater dietary diversity due to greater inter-individual variation in diet. In Saffron Finch, which delays plumage maturation, competition-driven partitioning of food resources seems essential in driving carotenoid-based plumage coloration between age classes and sexes.

The Impact of Different Surface Treatments on Topography and Bond Strength of Resin Cement to Lithium Disilicate Glass Ceramic.

OBJECTIVE: To evaluate the influence of different surface treatments on topography, surface roughness, surface energy, and microtensile bond strength stability of resin cement to lithium disilicate glass ceramic. METHODS AND MATERIALS: Seventy disc-shaped specimens of IPS e.max Press were divided into seven groups according to the surface treatment: NT - non-treated (control); FSil - hydrofluoric acid (HF) + silane; FPSil - HF + phosphoric acid (HP) + silane; FUSil - HF + ultrasonic bath (U) + silane; FPUSil - HF + HP + U + silane; MEP - Monobond Etch and Prime (MEP); and MEPH - MEP + heating. Topography and surface roughness were evaluated using a 3D laser confocal microscope (3DLCM) and scanning electron microscopy (SEM) and surface energy with a goniometer. The microtensile bond strength (µTBS) was evaluated after storage in distilled water at 37°C for 24 hours and after thermocycling (5,000 cycles, 5°-55° C, 30-second dwell time). Data were analyzed using one-way ANOVA (surface roughness and surface energy), two-way ANOVA (µTBS), Tukey's HSD post-hoc test, and Student t-test (α=0.05). RESULTS: FUSil, FPSil, and FSil presented similar and highest surface roughness, whereas NT, FPUSil, MEP, and MEPH showed similar, and lowest, roughness values (p<0.05). FPUSil, FPSil, FUSil, and FSil presented a similar and highest surface energy. NT, MEP, and MEPH showed similar and lowest surface energy. CONCLUSION: Only FSil, FPSil, and MEPH maintained bond strength stability after thermocycling, with FPSil leading to less bond strength reduction, suggesting this protocol is more reliable for bonding resin cements to lithium disilicate glass ceramics.

Seasonal variation of natural mortality factors of the guava psyllid Triozoida limbata.

It is important to understand how components of the agroecosystem interfere with the attack of a pest species and their seasonality in order to use these components in IPM programs. This study focused on the evaluation of the seasonality of natural control factors associated with the guava psyllid Triozoida limbata (Enderlein) in Brazil. Life-table data were collected from an experimental guava orchard during four periods that roughly represented four seasons. Natural mortality was monitored daily through the immature stages, and the relative importance of each natural mortality factor and its seasonality was determined. Significant statistical differences were observed in the mortality during the four periods (P < 0.05). Several factors contributed to the mortality of T. limbata, including rainfall, physiological disturbance, the parasitoid Psyllaephagus sp. (Hymenoptera: Encyrtidae) and specific predators: syrphids, predatory wasps and other generalist predators. Depending on the location of nymphs (exposed or inside galls), the relative importance of the different natural mortality factors changed. The principal component analysis (PCA) showed some trends in the relationship of natural control agents and weather conditions. For example, the occurrence of predatory wasps was positively correlated with temperature and occurrence of winds; the occurrence of syrphids and Psyllaephagus sp. were negatively correlated with temperature and winds; and the occurrence of other generalist predators were negatively correlated with the occurrence of rainfall and photoperiod. The results showed the importance of natural mortality factors for the management of T. limbata and their changes through the different seasons which should be considered when implementing IPM programs in guava orchards.

Functional and structural parameters to assess the ecological status of a metal contaminated area in the tropics.

Ecological parameters (soil invertebrates, microbial activity, and plant community) were assessed in a metal contaminated site in an abandoned lead smelter and non-contaminated reference sites, as part of an ecological risk assessment (ERA). Vegetation cover inside the smelter area was lower and presented a more homogenous species composition than outside. A more simplified and less abundant vegetation community within the smelter area also simplified the habitat conditions, which in addition to metal toxicity, impaired the soil microbial and faunal communities. A significant reduction in the feeding activity was observed within the smelter area. Also a significant change in community composition of surface dwelling invertebrates was observed at those sites when compared to sites outside the smelter area. Moreover, basal respiration, microbial biomass C, dehydrogenase and phosphatase activity also decreased in several of these points under the smelter area. As a result, a significant impairment of organic material decomposition in the most contaminated sites was observed. Metal contamination affected the ecological status of the site, leading to a risk for ecosystem functioning and provisioning of ecosystem services like organic matter decomposition and nutrient cycling, even 17 years after the end of smelting activities. Regarding the sensitivity of the ecological parameters assessed, most were able to distinguish sites within the smelter area boundaries from those outside. However, only bait lamina (feeding activity), basal respiration and microbial biomass carbon presented high capacity to distinguish the level of soil contamination, since they were significantly correlated with metal loadings, and thus are promising candidates to be integrated in the Ecological Line of Evidence of an ERA.

Copaiba oil and Neem extract can be a potential alternative for the behavioral control of Sitophilus zeamais.

Insects' ethology is an important factor when it is desired to carry out pest management. This knowledge makes it possible to manipulate behavioral activities, repel, or attract insects according to needs and interests. The maize weevil Sitophilus zeamais (Mots., 1855) (Coleoptera: Curculionidae), one of the main stored grain pests, has been the target of studies of behavioral changes studies through natural substances due to its resistance to different insecticidal classes. Thus, this study aimed to evaluate the effect of sublethal concentrations of neem extract and copaiba oil on the locomotor behavior of S. zeamais. The behavioral characteristic considered were walking activity, the frequency of contact of insects with the treated grain mass, and the time spent for this behavior. The walking activity of the S. zeamais increased with exposure to Neem extract and Copaiba oil. In general, the Neem extract and Copaiba oil-induced more contact with grain mass than the control, suggesting an attractive effect on the insect, however more significant for the Neem oil. The insect's behavior was altered, presenting a specific path due to Copaiba oil and Neem extract stimuli. These results indicate that Copaiba oil and Neem extract can be a potential alternative for controlling S. zeamais on stored products since changes in this pests' behavior can reduce qualitative and quantitative grain damage. Thus, the development of products based on Copaiba oil and Neem extract may be helpful for storage pest management.

Cardiorespiratory and neuromuscular responses during water aerobics exercise performed with and without equipment.

The aim of the study was to compare the cardiorespiratory and neuromuscular responses to water aerobics exercise performed with and without equipment. 15 women performed stationary jogging combined with elbow flexion/extension without equipment, with water-drag forces equipment and with water-floating equipment, at 2 submaximal cadences and at maximal cadence. Heart rate, oxygen uptake and electromyographic signal from biceps brachii, triceps brachii, biceps femoris and rectus femoris were collected during the exercise. The heart rate and oxygen uptake showed significantly higher values during the execution of the water aerobics exercise with either equipment compared to the execution without equipment. In addition, significant difference was found between submaximal cadences. For neuromuscular responses, no significant differences were found between the submaximal cadences for all muscles analyzed; however, significant differences were found between these submaximal cadences and the maximal cadence. Similarly, the results showed no significant differences between the execution of the exercise with or without equipment, except in the muscle activation of triceps brachii and biceps femoris, which was higher when using water-floating and water-drag forces equipment, respectively. In conclusion, the water aerobics exercise presented higher cardiorespiratory responses with equipment and also increased the cadence of execution. Nevertheless, neuromuscular responses were higher only at maximal cadence.

Modulatory effects of Duguetia furfuracea (A. St. Hil) Benth. and Hook. f. in Drosophila melanogaster somatic and germinative cells.

Mutagenic and antimutagenic activities of the medicinal plant Duguetia furfuracea were assessed using SMART/wing and ring-X-loss tests. For the ring-X-loss test, 2- to 3-day-old Drosophila melanogaster ring-X-lineage males and virgin ywsn³ females received D. furfuracea infusion at doses of 0.085, 0.042, or 0.014 g/mL for 24 h. We found that D. furfuracea did not produce any mutagenic effects in D. melanogaster germinative cells. The somatic cells of D. melanogaster were analyzed using the SMART/wing test involving three lineages - mwh, flr³, and ORR - and the same doses of D. furfuracea infusion employed in the ring-X-loss test, as well as 20 mM urethane. The results of both standard (ST) and high bioactivation (HB) crosses showed absence of mutagenic activity of D. furfuracea. In contrast, in both ST and HB crosses, we observed a modulatory effect of D. furfuracea against the genotoxic activity of urethane.

Mutagenic potential and modulatory effects of the medicinal plant Luehea divaricata (Malvaceae) in somatic cells of Drosophila melanogaster: SMART/wing.

Luehea divaricata is a native plant of the Brazilian Cerrado, known as "açoita-cavalo". It is used as a popular herbal medicine in the treatment of dysentery, bleeding, arthritis, tumors, ulcers, and gangrenous wounds. Considering that herbal medicines sometimes provoke tumors and/or may prevent mutational events, it is important to study the action of these natural drugs on DNA. Aqueous extract of the bark of L. divaricata was evaluated at three different concentrations (0.10, 0.30, 0.50 mg/mL), individually and in combination with the neoplastic drug doxorubicin (DXR), by the somatic mutation and recombination test (SMART/wing) in Drosophila melanogaster. Distilled water was included as a negative control. The mutation frequency in the treatments with L. divaricata extract alone was not significantly higher than in the negative control for standard (ST) and high bioactivation (HB) crosses. When L. divaricata extract was combined with DXR, there was a significant reduction in the frequency of spots when compared to DXR alone, in both crosses. Further studies with other experimental models would be useful to confirm that L. divaricata extract is not harmful and that it could be used in the prevention of cancer.

Providing inoculum for Didymella bryoniae studies: the effect of light spectrum and storing at low temperature.

The in vitro sporulation of Didymella bryoniae is of great importance for studies that require pure inoculum and in large quantities. Thus, the objectives of this study were to identify the best condition for D. bryoniae sporulation combining different light spectra (UV-A or UV-B light, white light, and continuous dark), with distinct culture media (PDA, V8, ML, and PDAB) and, to evaluate fungus' survivability stored at -20°C over time. The fungus samples were only able to sporulate when subjected to the UV-B light treatment, regardless of the culture medium. The highest appearance of spores conidium type was observed in the PDAB medium, and the lowest production occurred in the ML medium. Reproductive structures, such as perithecia and pycnidia, were observed in all culture media. However, there was considerable variation in the amount of each structure between the different culture media. The ML and V8 media showed a greater number of perithecia and the PDA and PDAB media presented a greater proportion of pycnidia compared to perithecia. The storage duration at -20°C did not affect mycelial growth or mycelial growth rate. In conclusion, the UV-B light is essential for D. bryoniae in vitro sporulation. Moreover, the culture medium composition influences the type of fungal structure produced, as well as spores' size and quantity. Freezing at -20°C is an efficient technique that can be used to store D. bryoniae for at least five months without loss of viability.