The Novel Mas agonist,CGEN-856S,Attenuates Isoproterenol-Induced Cardiac Remodeling and Myocardial Infarction Injury in Rats

CGEN-856S is a novel Mas agonist. Herein, we examined the effects of this peptide on isoproterenol (ISO)-induced cardiac remodeling and myocardial infarction (MI) injury. We also sought to determine whether CGEN-856S activates the underlying mechanisms related to Mas receptor activation. Heart hypertrophy and fibrosis were induced by ISO (2 mg·kg⁻¹·day⁻¹) in Wistar rats. After a 7-day treatment period with CGEN-856S (90 µg·kg⁻¹·day⁻¹) or vehicle, the cardiomyocyte diameter was evaluated in left ventricular sections stained with hematoxylin and eosin, and immunofluorescence labeling and quantitative confocal microscopy were used to quantify the deposition of type I and III collagen and fibronectin in the left ventricles. MI was induced by coronary artery ligation, and CGEN-856S (90 µg·kg⁻¹·day⁻¹) or saline was administered for 14 days. The Langendorff technique was used to evaluate cardiac function, and left ventricular sections were stained with Masson’s trichrome dye to quantify the infarct area. Using Chinese hamster ovary cells stably transfected with Mas cDNA, we evaluated whether CGEN-856S alters AKT and endothelial nitric oxide synthase (eNOS) phosphorylation. CGEN-856S reduced the degree of ISO-induced hypertrophy (13.91±0.17 µm vs. 12.41±0.16 µm in the ISO+CGEN-856S group). In addition, the Mas agonist attenuated the ISO-induced increase in collagen I, collagen III, and fibronectin deposition. CGEN-856S markedly attenuated the MI-induced decrease in systolic tension, as well as in +dT/dt and -dT/dt. Furthermore, CGEN-856S administration significantly decreased the infarct area (23.68±2.78% vs. 13.95±4.37% in the MI+CGEN-856S group). These effects likely involved the participation of AKT and NO, as CGEN-856S administration increased the levels of p-AKT and p-eNOS. Thus, our results indicate that CGEN-856S exerts cardioprotective effects on ISO-induced cardiac remodeling and MI-mediated heart failure in rats through a mechanism likely involving the eNOS/AKT pathway.     

Corneal Absorption of a New Riboflavin-Nanostructured System for Transepithelial Collagen Cross-Linking

Corneal collagen cross-linking (CXL) has been described as a promising therapy for keratoconus. According to standard CXL protocol, epithelium should be debrided before treatment to allow penetration of riboflavin into the corneal stroma. However, removal of the epithelium can increase procedure risks. In this study we aim to evaluate stromal penetration of a biocompatible riboflavin-based nanoemulsion system (riboflavin-5-phosphate and riboflavin-base) in rabbit corneas with intact epithelium. Two riboflavin nanoemulsions were developed. Transmittance and absorption coefficient were measured on corneas with intact epithelia after 30, 60, 120, 180, and 240 minutes following exposure to either the nanoemulsions or standard 0.1% or 1% riboflavin-dextran solutions. For the nanoemulsions, the epithelium was removed after measurements to assure that the riboflavin had passed through the hydrophobic epithelium and retained within the stroma. Results were compared to de-epithelialized corneas exposed to 0.1% riboflavin solution and to the same riboflavin nanoemulsions for 30 minutes (standard protocol). Mean transmittance and absorption measured in epithelialized corneas receiving the standard 0.1% riboflavin solution did not reach the levels found on the debrided corneas using the standard technique. Neither increasing the time of exposure nor the concentration of the riboflavin solution from 0.1% to 1% improved riboflavin penetration through the epithelium. When using riboflavin-5-phosphate nanoemulsion for 240 minutes, we found no difference between the mean absorption coefficients to the standard cross-linking protocol (p = 0.54). Riboflavin nanoemulsion was able to penetrate the corneal epithelium, achieving, after 240 minutes, greater stromal concentration when compared to debrided corneas with the standard protocol (p = 0.002). The riboflavin-5-phosphate nanoemulsion diffused better into the stroma than the riboflavin-base nanoemulsion.     

Root colonization and spore abundance of arbuscular mycorrhizal fungi in distinct successional stages from an Atlantic rainforest biome in southern Brazil

The influence of plant functional groups and moderate seasonality on arbuscular mycorrhizal (AM) fungal status (root colonization and spore density) was investigated during 13 consecutive months in a chronosequence of succession in southern Brazil, consisting of grassland field, scrub vegetation, secondary forest and mature forest, in a region of transition from tropical to subtropical zones. AM root colonization and spore density decreased with advancing succession and were highest in early successional sites with grassland and scrub vegetation, intermediary in the secondary forest and lowest in the mature forest. They were little influenced by soil properties, but were sufficiently influenced by the fine root nutrient status and fine root traits among different functional plant groups. AM root colonization and spore density were higher during the favourable plant growth season (spring and summer) than during the less favourable plant growth season (autumn and winter). Spore density displayed significant seasonal variation at all sites, whilst root colonization displayed significant seasonal variation in grassland, scrub and secondary forest, but not in mature forest. The data suggest that (1) different plant functional groups display different relationships with AM fungi, influencing their abundance differentially; (2) plant species from early successional phases are more susceptible to AM root colonization and maintain higher AM sporulation than late successional species; (3) fine root traits and nutrient status influence these AM fungal attributes; and (4) higher AM spore production and root colonization is associated with the season of higher light incidence and temperature, abundant water in soil and higher plant metabolic activity.     

Transglutaminase 2 as a biomarker of osteoarthritis: an update

Osteoarthritis is a progressive joint disease characterized by cartilage degradation and bone remodelling. Under physiologic conditions, articular cartilage displays a stable chondrocyte phenotype, whereas in osteoarthritis a chondrocyte hypertrophy develops near the sites of cartilage surface damage and associates to the pathologic expression of type X collagen. Transglutaminases (TGs) include a family of Ca²⁺-dependent enzymes that catalyze the formation of γ-glutamyl cross-links. Their substrates include a variety of intracellular and extracellular macromolecular components. TGs are ubiquitously and abundantly expressed and implicated in a variety of physiopathological processes. TGs activity is modulated by inflammatory cytokines. TG2 (also known as tissue transglutaminase) mediates the hypertrophic differentiation of joint chondrocytes and interleukin-1-induced calcification. Histomorphometrical and biomolecular investigations document increased TG2 expression in human and experimental osteoarthritis. Consequently, the level of TG2 expression may represent an adjuvant additional marker to monitor tissue remodelling occurring in osteoarthritic joint tissue. Experimental induction of osteoarthritis in TG2 knockout mice is followed from reduced cartilage destruction and increased osteophyte formation compared to wild-type mice, suggesting a different influence on joint bone and cartilage remodelling. The capacity of transamidation by TG2 to regulate activation of latent TGF-β seems to have a potential impact on the regulation of inflammatory response in osteoarthritic tissues. Additional studies are needed to define TG2-regulated pathways that are differently modulated in osteoblasts and chondrocytes during osteoarthritis.     

Diurnal Variation in Scent Marking Behavior in Captive Male and Female Common Marmosets,Callithrix jacchus

Chemical communication by scent-marking behavior in New World primates is used to prevent the access of potential competitors to a territory, to identify food resources and the reproductive condition of mates, among others. In common marmosets, primates of the Callitrichidae family, this behavior also occurs as olfactory identification of an individual or of the reproductive status of females. Despite this information, the diurnal variation and gender differences in the profile of this behavior remain to be investigated. The aims of this study were to establish the diurnal profile of the distribution of this behavior and the influence of the sex of markers. We used 18 adult common marmosets, Callithrix jacchus, 10 males and 8 females from 6 family groups (6 fathers and 4 sons; 4 mothers and 4 daughters). The frequency of scent-marking behavior was recorded for each animal over a period of 8 days, twice a week, for 4 weeks, starting when the animals left the nest box (approximately at 05:00 a.m.) until the end of the photophase, at about 05:00 p.m. A MANOVA test was performed to compare the frequency of scent-marking behavior at 2 hour intervals using pooled data for males and females. The results showed that significantly higher levels of scent-marking behavior occurred during the 03:00–05:00 p.m. interval compared to all other intervals. Lower values were recorded during the 11:00–13:00 interval and an effect of the sex factor was also found, with the values being higher for females than for males, although a significant difference was recorded only for the 07:00–09:00 interval. Minimal values for males were recorded during the 07:00–09:00 interval, whereas minimum values for females were recorded during the 11:00–13:00 interval. However, the highest values for both sexes continued to occur during the 15:00–17:00 interval. These results suggest that scent marking behavior in common marmosets has a preferential incidence at the end of the day and this might be occurring in association with feeding behavior. At this time these animals usually forage more to prepare for the night's fast. Since these animals can discriminate chemical clues as long as 24 hours after they have been left, the higher incidence of this behavior at this time probably will assure that the animals will localize feeding resources used on the preceding day. Significant elevation of scent marking behavior in females in relation to males was found only at 07:00–09:00 interval and seems to be associated with signalizing of reproductive status, preferential access to foraging or both.     

Inactivating mutation in histone deacetylase 3 stabilizes its active conformation

Histone deacetylases (HDACs), together with histone acetyltransferases (HATs), regulate gene expression by modulating the acetylation level of chromatin. HDAC3 is implicated in many important cellular processes, particularly in cancer cell proliferation and metastasis, making inhibition of HDAC3 a promising epigenetic treatment for certain cancers. HDAC3 is activated upon complex formation with both inositol tetraphosphate (IP4) and the deacetylase‐activating domain (DAD) of multi‐protein nuclear receptor corepressor complexes. In previous studies, we have shown that binding of DAD and IP4 to HDAC3 significantly restricts its conformational space towards its stable ternary complex conformation, and suggest this to be the active conformation. Here, we report a single mutation of HDAC3 that is capable of mimicking the stabilizing effects of DAD and IP4, without the presence of either. This mutation, however, results in a total loss of deacetylase activity, prompting a closer evaluation of our understanding of the activation of HDAC3.     

Triploidization in the streaked prochilod Prochilodus lineatus inferred by flow cytometry,blood smears and karyological approaches

The streaked prochilod Prochilodus lineatus is an important fish species from the Neotropical region. In this study, a protocol of triploidization was established by temperature shock. Fertilized eggs were shocked at 2 min post-fertilization at 0, 38, 40 and 42°C. At 0°C, the embryos were maintained for 30 min, while the rest for 2 min. Heat shocked embryos and control (during all experiment) were incubated at 27°C. The ploidy status was confirmed by flow cytometry, erythrocyte diameter, conventional cytogenetics (Giemsa staining), chromosome banding and fluorescence in situ hybridization using 5S and 18S rDNA probes. Heat-shock at 40°C produced 96.7% of triploids and such a procedure did not reduce the fertilization, hatching rate and the percentage of normal embryos. The use of chromosome banding and FISH gave rise to effective procedure to identify triploids. The data obtained here is innovative for the species and bring new information for basic and applied studies.     

Chemical ecology of Cyclocephala forsteri (Melolonthidae), a threat to macauba oil palm cultivars (Acrocomia aculeata,Arecaceae)

The environmental unsustainability of African oil palm crops is a growing worldwide concern, and the macauba palm in tropical Southern America surges as the most viable substitute for the production of first-generation biodiesel as a renewable fuel. Large-scale cultivation of the macauba palm, however, is bound to suffer a major setback due to the fast emergence of a florivorous pest threat, Cyclocephala forsteri, a scarab beetle species that can severely reduce fruit yield. Like other congenerics, female and male C. forsteri are night-foraging and driven in large numbers to macauba palm inflorescences by specific volatile organic compounds. In the present study, we assessed the kairomonal cues involved in this plant–florivore interaction and their potential application in selective pest management strategies. Headspace samples of the floral fragrance of macauba palms at our study site in Central Brazil were largely dominated by 4-methylanisole (>97% relative amount), which along with the minor constituents 2-isopropyl-3-methoxypyrazine and 2-sec-butyl-3-methoxypyrazine triggered electroantennographic responses from both female and male C. forsteri. Field behavioural assays evidenced that beetles of this species were exclusively attracted to scented traps baited with 4-methylanisole. Increased total scent discharge attained with an ultrasonic piezoelectric diffuser has positively influenced attractiveness efficiency of the trapping setup. 4-Methylanisole is hereby identified as yet another volatile kairomone involved in highly selective attraction of potential pest florivorous cyclocephaline scarabs associated with economically exploitable palms and should be viewed as a key element in short-term integrated management plans for the cultivation of the macauba palm in South America.