Effects of frutalin and doxorubicin on growth, ultrastructure and gene expression in goat secondary follicles cultured in vitro.

This study evaluated the effects of frutalin (0.6, 6.0 or 60.0 µg/mL) and doxorubicin (0.3 µg/mL) on survival, growth and ultrastructure of in-vitro cultured goat secondary follicles. The effects of these substances on the levels of mRNA for Casp3, Casp6, Bax, and Bcl2 were also investigated. Results showed that, after 6 days of culture, frutalin or doxorubicin reduced the percentage of normal follicles (P < 0.05), but doxorubicin had higher toxicity than frutalin. Except for follicles cultured with 0.6 µg/mL frutalin, follicular growth rate was reduced after culture with doxorubicin or frutalin (P < 0.05). The presence doxorubicin or 60.0 µg/mL frutalin increased the levels of mRNA for Casp3, Casp6, Bax, and Bcl2 (P < 0.05). Higher mRNA levels for Casp3, Casp6 and Bax were found in follicles cultured with doxorubicin, but higher levels of Bcl2 mRNA were found in follicles cultured with frutalin (P < 0.05). In conclusion, frutalin has lower toxic effects than doxorubicin on secondary follicles cultured in vitro.

Dose-dependent effects of frutalin on in vitro maturation and fertilization of pig oocytes.

The aim of the present study was to evaluate the effect of frutalin (FTL) on in vitro maturation (IVM), and fertilization (IVF) of pig oocytes. In the Experiment 1, cumulus-oocyte complexes (COCs) were submitted to IVM in maturation medium alone or supplemented with different FTL concentration (0.6, 6 and 60 µg/mL), or 0.3 µg/mL doxorubicin (DXR). After IVM, some oocytes were evaluated for chromatin configuration, and the remaining oocytes were submitted to in vitro fertilization. In Experiment 2, matured oocytes were fertilized in IVF medium alone (control) or in presence of different FTL concentration (0.6, 6 and 60 µg/mL), or 0.3 µg/mL DXR. After 18 h post fertilization, the endpoints penetration rate, monospermy, spermatozoa per oocyte, and the IVF efficiency were evaluated in both experiments. In Experiment 1, 6 and 60 µg/mL FTL, as well as DXR increased (P < 0.05) the rate of oocytes with abnormal chromatin configuration when compared to oocyte matured in control medium alone or supplemented with 0.6 µg/mL FTL. The percentage of meiotic resumption in oocytes cultured with 60 µg/mL FTL or DXR was less (P < 0.05) than in the other treatments. Moreover, oocytes matured with 6 or 60 µg/mL FTL and DXR had a lesser IVM efficiency when compared to those matured with 0.6 µg/mL FTL or in control medium. Additionally, there was a greater (P < 0.05) with culture in a medium containing 6 µg/mL FTL for the rate of partenogenetically activated oocytes when compared with the other treatments. Culturing of COCs during IVM in a medium containing 6 or 60 FTL resulted in a lesser (P < 0.05) sperm penetration and spermatozoa/oocyte rates when compared to other treatments, and IVF efficiency was less (P < 0.05) than that in control medium alone or with a medium containing 0.6 µg/mL FTL. In Experiment 2, culturing in a medium containing 0.6 µg/mL FTL resulted in greater (P < 0.05) monospermy and IVF efficiency rates when compared to culturing in the control medium. In addition, culturing in a medium with 6 and 60 µg/mL FTL resulted in a lesser (P < 0.05) spermatozoa penetration, sperm/oocyte rates and IVF efficiency, although there were greater (P < 0.05) monospermy rates. In conclusion, culturing in a medium containing 0.6 µg/mL FTL resulted in lesser spermatozoa penetration rates and number of spermatozoa/oocyte increasing the IVF efficiency without harmful effects. Use of a greater concentration of FTL in the medium has toxic effects during oocyte maturation and results in a reduced IVF efficiency.

Effect of oral ingestion of an extract of the herb Uncaria tomentosa on the biodistribution of sodium pertechnetate in rats.

The aim of the present study was to determine the effect of the oral ingestion of an extract of the herb Uncaria tomentosa (cat's claw) on the biodistribution of the radiobiocomplex sodium pertechnetate (Na99mTcO4) in rats. The animals (male Wistar rats, 2 months old, 180-220 g), were treated (1 mL) with an U. tomentosa extract (32 mg/mL, N = 5) or 0.9% NaCl solution (control, N = 5) for 7 days. After this period, Na99mTcO4 (3.7 MBq, 0.3 mL) was injected through the ocular plexus and after 10 min the rats were killed, the organs isolated and counted in a well-gamma counter. A significant (P < 0.05) alteration in Na99mTcO4 uptake i) from 0.57 +/- 0.008 to 0.39 +/- 0.06 %ATI/organ (P < 0.05) and from 0.57 +/- 0.17 to 0.39 +/- 0.14 %ATI/g (P < 0.05) was observed in the heart, ii) from 0.07 +/- 0.02 to 0.19 +/- 0.07 %ATI/g in the pancreas, and iii) from 0.07 +/- 0.01 to 0.18 +/- 0.07 %ATI/g (P < 0.05) in muscle after treatment with this extract. Although these results were obtained with animals, caution is advisable in the interpretation of the nuclear medicine examination when the patient is using this herb. This finding is probably an example of drug interaction with a radiopharmaceutical, a fact that could lead to misdiagnosis of the examination in clinical practice with unexpected consequences for the patient.

Effect of oral ingestion of an extract of the herb Uncaria tomentosa on the biodistribution of sodium pertechnetate in rats

The aim of the present study was to determine the effect of the oral ingestion of an extract of the herb Uncaria tomentosa (cat's claw) on the biodistribution of the radiobiocomplex sodium pertechnetate (Na99mTcO4) in rats. The animals (male Wistar rats, 2 months old, 180-220 g), were treated (1 mL) with an U. tomentosa extract (32 mg/mL, N = 5) or 0.9 percent NaCl solution (control, N = 5) for 7 days. After this period, Na99mTcO4 (3.7 MBq, 0.3 mL) was injected through the ocular plexus and after 10 min the rats were killed, the organs isolated and counted in a well-gamma counter. A significant (P < 0.05) alteration in Na99mTcO4 uptake i) from 0.57 ± 0.008 to 0.39 ± 0.06 percentATI/organ (P < 0.05) and from 0.57 ± 0.17 to 0.39 ± 0.14 percentATI/g (P < 0.05) was observed in the heart, ii) from 0.07 ± 0.02 to 0.19 ± 0.07 percentATI/g in the pancreas, and iii) from 0.07 ± 0.01 to 0.18 ± 0.07 percentATI/g (P < 0.05) in muscle after treatment with this extract. Although these results were obtained with animals, caution is advisable in the interpretation of the nuclear medicine examination when the patient is using this herb. This finding is probably an example of drug interaction with a radiopharmaceutical, a fact that could lead to misdiagnosis of the examination in clinical practice with unexpected consequences for the patient.

NADPH-diaphorase activity in area 17 on the squirrel monkey visual cortex: neuropil pattern, cell morphology and laminar distribution

We studied the distribution of NADPH-diaphorase activity in the visual cortex of normal adult New World monkeys (Saimiri sciureus) using the malic enzyme "indirect" method. NADPH-diaphorase neuropil activity had a heterogeneous distribution. In coronal sections, it had a clear laminar pattern that was coincident with Nissl-stained layers. In tangential sections, we observed blobs in supragranular layers of V1 and stripes throughout the entire V2. We quantified and compared the tangential distribution of NADPH-diaphorase and cytochrome oxidade blobs in adjacent sections of the supragramular layers of V1. Although their spatial distributions were rather similar, the two enzymes did not always overlap. The histochemical reaction also revealed two different types of stained cells: a slightly stained subpopulation and a subgroup of deeply stained neurons resembling a Golgi impregnation. These neurons were sparsely spined non-pyramidal cells. Their dendritic arbors were very well stained but their axons were not always evident. In the gray matter, heavily stained neurons showed different dendritic arbor morphologies. However, most of the strongly reactive cells lay in the subjacent white matter, where they presented a more homogenous morphology. Our results demonstrate that the pattern of NADPH-diaphorase activity is similar to that previously described in Old World monkeys.