Decreasing from 9 to 7 days the permanence of progesterone inserts make possible their use up to 5 folds in suckled Nellore cows.

This study aimed to evaluate the effects of decreasing permanence period of progesterone (P4) inserts from 9 (9d) to 7 (7d) days in timed-AI (TAI) protocol, as well as their reuse on pregnancy per AI (P/AI) during 7d protocol. At the beginning of all protocols, cows received 2 mg of estradiol benzoate (EB) and a vaginal insert containing 1.9 g of P4. In the 7d protocol, the P4 insert was removed, and cows were given 25 mg of prostaglandin F2α, 0.6 mg of estradiol cypionate and 300 IU of eCG 7 days later. In the 9d protocol, the P4 insert was removed, and cows received the same hormones that were administrated in 7d protocol, however, they were applied on the 9th day of the protocol. In the experiment I, 302 suckled Nellore cows were undergone to 7d protocol, and AI was performed 10-14 h later after estrus detection. In the experiment II, 679 suckled Nellore cows were assigned to receive either 7d or 9d protocols. In the experiment III, 999 suckled Nellore cows were assigned to receive either a new P4 insert (CIDR1), or a P4 insert used previously for 7 (CIDR2), 14 (CIDR3), 21 (CIDR4) or 28 (CIDR5) days, and 227 Nellore heifers received a P4 insert used previously for 21 (CIDR4) or 35 (CIDR6) days during the 7d protocol. When 7d protocol was used, 45% of cows exhibited estrus 48 h after P4 removal. Thus, the AI was performed 55 h after P4 removal in experiments II and III when 7d protocol was used. There was no difference in estrus detection rate (72 vs 74%; P = 0.60), ovulation rate (80 vs 88%; P = 0.13) and P/AI (56 vs 54%; P = 0.49) between 7d and 9d protocols, respectively. In the 7d protocol, the P/AI was similar (P = 0.72) using a new P4 insert (47%), or a P4 insert used previously for 7 (48%), 14 (45%), 21 (54%), or 28 (49%) days in Nellore suckled cows. In addition, P/AI was similar (P = 0.15) in heifers that received a P4 insert used previously for 21 (52%), or 35 (61%) days during the 7d protocol. In conclusion, cows submitted to 7d or 9d protocols had similar reproductive performance, and the reuse of P4 inserts up to 6 folds (five in suckled cows plus one in heifers) did not affect reproductive performance of Nellore cattle in 7d protocol.

Natural polyprenylated benzophenone: keto-enol tautomerism from density functional calculations and the AIM theory.

The quantum theory of atoms in molecules (QTAIM) and density functional theory (DFT) calculations were employed to investigate the structure and tautomeric equilibrium of epiclusianone, a polyisoprenylated benzophenone with interesting biological activities. Two different exchange-correlation functionals were employed, namely ωB97x-D and M06-2x, including implicit solvent models (benzene and DMSO). Our results for the thermodynamic properties show that the isomer in which the H atom is bonded to the oxygen away from the benzene ring is the most stable tautomer form of the epiclusianone, thus confirming previous charge density analysis from X-ray diffraction data (Martins et al. J Braz Chem Soc 18(8):1515-1523, 22).

Comparison of two timed artificial insemination system schemes to synchronize estrus and ovulation in Nellore cattle.

The aim of this study was to evaluate the reproductive performance of 411 Nellore cows (198 nulliparous, 80 primiparous, and 133 multiparous) submitted to the 5dCO-Synch + P4 or 7dEB + P4 systems. The 5dCO-Synch + P4 system consisted of insertion of an intravaginal progesterone (P4) insert and 100 µg of GnRH (intramuscularly [i.m.]) on Day 0. On Day 5, the P4 insert was removed, and two doses of 25 mg of PGF2α (i.m.) were administered 6 hours apart. Cows not detected in estrus until 55 hours after insert removal received 100 µg of GnRH i.m. 17 hours later (i.e., 72 hours after P4 removal). The 7dEB + P4 system consisted of insertion of a P4 insert and 2 mg of estradiol benzoate (i.m.) on Day 0. On Day 7, the P4 insert was removed and 25 mg of PGF2α, 0.6 mg of estradiol cypionate, and 300 IU of eCG were administered i.m. In both systems, artificial insemination (AI) was performed according to estrus detection (i.e., cows detected in estrus until 55 hours after insert removal were inseminated at 55 hours and cows detected in estrus later or those not detected in estrus were inseminated at 72 hours). Estrus-detection risk was greater (P < 0.05) in 7dEB + P4 (80.4%) than 5dCO-Synch + P4 system (36.4%). Progesterone concentration 10 days after AI was greater (P < 0.05) in 7dEB + P4 than 5dCO-Synch + P4 system in primiparous and multiparous but did not differ between systems in nulliparous cows. Pregnancy per AI was greater (P < 0.05) in 7dEB + P4 (49.7%) than 5dCO-Synch + P4 (35.4%) system. Primiparous had lower estrus-detection risk (25.0%), ovulation risk (76.6%), and pregnancy per AI (28.7%) than multiparous or nulliparous cows. In conclusion, reproductive performance was reduced with the 5dCO-Synch + P4 in comparison with the 7dEB + P4 system in Nellore cows. Moreover, the reproductive traits observed for primiparous cows indicate that more attention is required when timed AI programs are started early after calving.

The natural flavone fukugetin as a mixed-type inhibitor for human tissue kallikreins.

The human tissue kallikreins (KLK1-KLK15) comprise a family of 15 serine peptidases detected in almost every tissue of the human body and that actively participate in many physiological and pathological events. Some kallikreins are involved in diseases for which no effective therapy is available, as for example, epithelial disorders, bacterial infections and in certain cancers metastatic processes. In recent years our group have made efforts to find inhibitors for all kallikreins, based on natural products and synthetic molecules, and all the inhibitors developed by our group presented a competitive mechanism of inhibition. Here we describe fukugetin, a natural product that presents a mixed-type mechanism of inhibition against KLK1 and KLK2. This type of inhibitor is gaining importance today, especially for the development of exosite-type inhibitors, which present potential to selectively inhibit the enzyme activity only against specific substrate.

7-Epiclusianone, a Benzophenone Extracted from Garcinia brasiliensis (Clusiaceae), Induces Cell Cycle Arrest in G1/S Transition in A549 Cells.

Lung cancer is the leading cause of cancer deaths in the world. Disease stage is the most relevant factor influencing mortality. Unfortunately, most patients are still diagnosed at an advanced stage and their five-year survival rate is only 4%. Thus, it is relevant to identify novel drugs that can improve the treatment options for lung cancer. Natural products have been an important source for the discovery of new compounds with pharmacological potential including antineoplastic agents. We have previously isolated a prenylated benzophenone (7-epiclusianone) from Garcinia brasiliensis (Clusiaceae) that has several biological properties including antiproliferative activity against cancer cell lines. In continuation with our studies, the present work aimed to investigate the mechanisms involved with antiproliferative activity of 7-epiclusianone in A549 cells. Our data showed that 7-epiclusianone reduced the viability of A549 cells in a concentration-dependent manner (IC50 of 16.13 ± 1.12 µM). Cells were arrested in G1/S transition and apoptosis was induced. In addition, we observed morphological changes with cytoskeleton disorganization in consequence of the treatment. Taken together, the results showed that cell cycle arrest in G1/S transition is the main mechanism involved with antiproliferative activity of 7-epiclusianone. Our results are promising and open up the prospect of using this compound in further anticancer in vivo studies.

Oleanane-type triterpenoid: an anti-inflammatory compound of the roots Arrabidaea brachypoda

AbstractArrabidaea brachypoda Bureau, Bignoniaceae, known as "cipó-una", is widely used in traditional medicine in Southeastern and Northeastern Brazil for kidney stones and painful joints. This study was aimed at evaluating the anti-inflammatory proprieties of the oleanane-type triterpenoid 3β-estearioxy-olean-12-ene isolated from the roots of A. brachypoda. Carrageenan-induced paw oedema, formalin test and hot plate test were used to investigate the antiinflammatory activity of 3β-estearioxy-olean-12-ene in animals. We observed that 3β-estearioxy-olean-12-ene at doses of 5, 10 and 15 mg/kg p.o. demonstrated anti-inflammatory effects, by reduced (p < 0.05) paw oedema induced by carrageenan and by decreased (p < 0.05) licking time caused by a subplantar injection of formalin. In conclusion, 3β-estearioxy-olean-12-ene, a triterpene isolated from the roots of A. brachypoda, demonstrate anti-inflammatory effect in different tests. Thus, it may be useful in the treatment of inflammatory disorders, which supports previous claims of its traditional use.

Anti-inflammatory and antinociceptive effects of Arrabidaea brachypoda (DC.) Bureau roots.

AIM OF THE STUDY: Arrabidaea brachypoda (DC.) Bureau has been used to relieve general pain, painful joints and kidney stones in Brazilian folk medicine. Nevertheless, scientific information regarding this species is scarce; there are no reports related to its possible analgesic and anti-inflammatory effects. This study was aimed at evaluating the traditional use of Arrabidaea brachypoda root using in vivo inflammatory and nociceptive models. MATERIALS AND METHODS: Carrageenan-induced paw edema, peritonitis and fibrovascular tissue growth induced by s.c. cotton pellet implantation were used to investigate the anti-inflammatory activity of Arrabidaea brachypoda roots ethanolic extract (AbEE) in rats. Formalin and acetic acid-induced writhing tests were used to investigate the antinociceptive activity in mice. High-performance liquid chromatography (HPLC) was used to determine the fingerprint chromatogram of AbEE. RESULTS: The AbEE at test doses of 30-300 mg/kg p.o. demonstrated anti-inflammatory effects. AbEE reduced paw edema induced by carrageenan, inhibited leukocyte recruitment into the peritoneal cavity and, in the model of chronic inflammation using the cotton pellet-induced fibrovascular tissue growth in rats, significantly inhibited the formation of granulomatous tissue. The extracts at test doses of 30-300 mg/kg p.o. clearly demonstrated antinociceptive activity, except during the first phase of the formalin test. The presence of quercetin and phenolic compounds in the extract Arrabidaea brachypoda was confirmed using HPLC. CONCLUSION: Arrabidaea brachypoda ethanol extract markedly demonstrated anti-inflammatory action in rats and antinociceptive activity in mice, which supports the previous claims of traditional use.

Anti-inflammatory and antinociceptive effects of Garcinia brasiliensis.

AIM OF THE STUDY: In Brazilian folk medicine, the leaves of Garcinia brasiliensis are used to treat tumors, inflammation of the urinary tract and arthritis as well as to relieve pain. Nevertheless, scientific information regarding Garcinia brasiliensis is limited; there are no reports related to its possible anti-inflammatory and antinociceptive effects. This study employed in vivo inflammatory and nociceptive models to evaluate the scientific basis for the traditional use of Garcinia brasiliensis. MATERIALS AND METHODS: Carrageenan-induced paw edema, peritonitis and fibrovascular tissue growth induced by s.c. cotton pellet implantation were used to investigate the anti-inflammatory activity of Garcinia brasiliensis ethanolic extract (GbEE) in rats. Formalin and acetic acid-induced writhing tests were used to investigate the antinociceptive activity in mice. RESULTS: GbEE at test doses of 30-300 mg/kg p.o. clearly demonstrated anti-inflammatory effects by reduced paw edema induced by carrageenan, inhibited leukocyte recruitment into the peritoneal cavity, and in the model of chronic inflammation using the cotton pellet-induced fibrovascular tissue growth in rats, the GbEE significantly inhibited the formation of granulomatous tissue. The extracts at test doses of 30-300 mg/kg, p.o., clearly demonstrated antinociceptive activity, except for the first phase of the formalin test. CONCLUSION: GbEE markedly demonstrated anti-inflammatory action in rats and antinociceptive activity in mice, which supports previous claims of the traditional use of species of the Garcinia genus for inflammation and pain.

A powder X-ray diffraction method for detection of polyprenylated benzophenones in plant extracts associated with HPLC for quantitative analysis.

A robust, direct, rapid and non-destructive X-ray diffraction crystallography method to detect the polyprenylated benzophenones 7-epi-clusianone (1) and guttiferone A (2) in extracts from Garcinia brasiliensis is presented. Powder samples of benzophenones 1 and 2, dried hexane extracts from G. brasiliensis seeds and fruit's pericarp, and the dried ethanolic extract from G. brasiliensis seeds were unambiguously characterized by powder X-ray diffractometry. The calculated X-ray diffraction peaks from crystal structures of analytes 1 and 2, previously determined by single-crystal X-ray diffraction technique, were overlaid to those of the experimental powder diffractograms, providing a practical identification of these compounds in the analyzed material and confirming the pure contents of the powder samples. Using the X-ray diffraction crystallography method, the studied polyprenylated benzophenones were selectively and simultaneously detected in the extracts which were mounted directly on sample holder. In addition, reference materials of the analytes were not required for analyses since the crystal structures of the compounds are known. High performance liquid chromatography analyses also were comparatively carried out to quantify the analytes in the same plant extracts showing to be in agreement with X-ray diffraction crystallography method.

Antiinflammatory properties of Morus nigra leaves.

The aim of the present study was to investigate antiinflammatory activity of the methylene chloride extract of Morus nigra in animal models. Carrageenan-induced paw edema as well as fibrovascular tissue growth induced by s.c. cotton pellet implantation were used to investigate the antiinflammatory activity of Morus nigra extract (MnE) in rats. A HPLC fingerprint was used for phytochemical analysis of the extracts. The MnE at test doses of 100-300 mg/kg p.o. clearly demonstrated antiinflammatory effects by reduced paw edema induced by carragenan and significantly inhibited the formation of granulomatous tissue. In addition, chemical compounds isolated from Morus nigra, including betulinic acid, ß-sitosterol and germanicol, may be responsible for the antiinflammatory effect of the extract.

Natural polyprenylated benzophenones inhibiting cysteine and serine proteases.

We have investigated the in vitro inhibition of papain, trypsin, and cathepsins B and G by five benzophenone-type compounds, three natural ones isolated from Garcinia brasiliensis and two synthetic derivatives. The activities of pentaprenylated trihydroxy-substituted benzophenone guttiferone A (1) on all assayed enzymes were approximately 2-69 folds higher than that manifested by mono-hydroxylated tetraprenylated and triprenylated compounds epiclusianone (2) and garciniaphenone (3), respectively, the other natural benzophenones that also inhibited significantly the four enzymes. Differently, the synthetic derivatives 2,2',4-trihydroxybenzophenone (4) and diphenylmethanone (5) have inhibited weakly the studied proteases. Furthermore, compound 1 has bonded preferentially to cathepsin G, once its IC(50) value (2.7+/-0.1 microM) on such peptidase is quite similar to that of the classical inhibitor of serine proteases, chymostatin (2.1+/-0.1 microM). Interesting structure-activity relationships (SARs) were confirmed by flexible docking simulations, likewise the potential usefulness of natural compound 1 as antitumoral drug is strengthened by our results concerning the antiproteolytic activity.

Composition, and anti-inflammatory and antioxidant activities of the volatile oil from the fruit peel of Garcinia brasiliensis.

The composition of the volatile oil obtained by hydrodistillation from the fruit peel of Garcinia brasiliensis (Mart.) Planch. et Triana was determined by GC/MS. A total of 38 components were identified, including gamma-muurolene (10.3%), spathulenol (8.7%), delta-cadinene (8.3%), torreyol (8.0%), alpha-cadinol (7.0%), cadalene (6.3%), and gamma-cadinene (5.3%). Oxygenated sesquiterpenes (43%) were the main group of compounds. The anti-inflammatory activity of the volatile oil was evaluated through the rat-paw edema model induced by carrageenan. Inhibition of the inflammatory process was noticed 3 h after carrageenan administration. In addition, the volatile oil showed poor antioxidant activity.