Follicular Development and Reproductive Characteristics in Four Species of Brazilian Tropidurus Lizards.

The follicular development and reproductive characteristics of four species of oviparous lizards in the Tropidurus torquatus group were anatomically and histologically evaluated. We measured specimens, recorded the number of follicles and eggs, and removed the right ovary of each individual, which we processed according to histological routine and photo-documented. For all species, ovaries were divided into a cortical germinal bed, where oogonia and stage I oocytes are located, and a medullar stroma, where the remaining follicular developmental stages occur. Microscopic analysis did not show differences in ovarian follicle development for the four species of the T. torquatus group. The only measurement that presented significant variation throughout follicular development was the thickness of the granulosa layer in stage VII follicles. Regarding snout-tovent length at sexual maturity, few variations were observed among the species, with the smallest length recorded for T. oreadicus. Clutch size was higher for T. itambere and T. torquatus species, with a maximum of five and six eggs in the oviducts, respectively. Tropidurus oreadicus and T. hispidus had a maximum of five and six follicles, respectively, but neither species presented eggs in the oviducts. In addition, the reproductive activity varied among the four lizard species of the T. torquatus group. Finally, besides the morphological characteristics observed among these species, this is the first study to report data on the germinal bed, number of ovarian follicles, corpus luteum, and follicular atresia in relation to reproductive activity.

Effect of 60 and 90 days of isotretinoin treatment on the structure of the small intestine mucosa in young male Wistar rats.

Isotretinoin is a substance used in cases of severe acne and acne resistant to other treatments. This skin disease affects patients of all ages and can interfere with social life, especially in adolescents. The drug acts by suppressing sebaceous gland activity and creating an inhospitable environment for Propionibacterium acne. The integrity of the small intestine is important for correct nutrition and patient treatment. We intended to assess the small intestine structure after treatment with 5 mg/kg isotretinoin solution and after a period without the drug, which could be considered a rest period. Young male Wistar rats (n=24) were separated into 4 groups (n=6): C: water; D0: soybean oil; D5a: 5 mg/kg; D5b: 5 mg/kg for 60 days followed by 30 days of rest period. Soybean oil was used to dilute the drug and it was offered daily by gavage. The animals were euthanized and the duodenum, jejunum and ileum were collected for analysis with light and scanning electron microscopy. The treatment stimulated tissue proliferation in the jejunum and ileum but had no significant effect in the duodenum. The results also showed a modification in goblet cell frequency in the duodenum and ileum. A further finding was that some modifications disappeared during the rest period. The protocol showed that the small intestine was somewhat altered by the treatment yet no lasting damage was caused.

Effectiveness of hyperbaric oxygen for experimental treatment of schistosomiasis mansoni using praziquantel-free and encapsulated into liposomes: assay in adult worms and oviposition.

The treatment of schistosomiasis depends on a single drug: praziquantel (PZQ). However, this treatment presents limitations such as low and/or erratic bioavailability that can contribute to cases of tolerance. Improvements to the available drug are urgently needed and studies with a controlled system of drug release, like liposomes, have been gaining prominence. The present study evaluated the activity and synergy between liposomal-praziquantel (lip.PZQ) and hyperbaric oxygen therapy (HBO). Mice received doses of 60 or 100mg/kg PZQ or lip.PZQ, 50 days post-infection, and after the treatment, were exposed to HBO (3 atmosphere absolute - ATA) for 1h. The viability of adult worms and oviposition were analyzed, by necropsy and Kato-Katz examination performed after 15 days of treatment. A concentration of 100mg/kg of lip.PZQ+HBO was more effective (48.0% reduction of worms, 83.3% reduction of eggs/gram of feces) and 100% of the mice had altered of oograms (indicating interruption of oviposition) compared to other treatments and to the Control group (infected and untreated). It is known that PZQ requires participation of the host immune system to complete its antischistosomal activity and that HBO is able to stimulate the immune system. The drug became more available in the body when incorporated into liposomes and, used with HBO, the HBO worked as an adjuvant. This explains the decreases of oviposition and worms recovered form hepatic portal system.

Comparative study of the bioconversion process using R-(+)- and S-(-)-limonene as substrates for Fusarium oxysporum 152B.

This study compared the bioconversion process of S-(-)-limonene into limonene-1,2-diol with the already established biotransformation of R-(+)-limonene into α-terpineol using the same biocatalyst in both processes, Fusarium oxysporum 152B. The bioconversion of the S-(-)-isomer was tested on cell permeabilisation under anaerobic conditions and using a biphasic system. When submitted to permeabilisation trials, this biocatalyst has shown a relatively high resistance; still, no production of limonene-1,2-diol and a loss of activity of the biocatalyst were observed after intense cell treatment, indicating a complete loss of cell viability. Furthermore, the results showed that this process can be characterised as an aerobic system that was catalysed by limonene-1,2-epoxide hydrolase, had an intracellular nature and was cofactor-dependent because the final product was not detected by an anaerobic process. Finally, this is the first report to characterise the bioconversion of R-(+)- and S-(-)-limonene by cellular detoxification using ultra-structural analysis.

Acupuncture increases the diameter and reorganisation of collagen fibrils during rat tendon healing.

BACKGROUND: Our previous study showed that electroacupuncture (EA) increases the concentration and reorganisation of collagen in a rat model of tendon healing. However, the ultrastructure of collagen fibrils after acupuncture is unknown. OBJECTIVES: To assess the effect of acupuncture protocols on the ultrastructure of collagen fibrils during tendon healing. METHODS: Sixty-four rats were divided into the following groups: non-tenotomised (normal group), tenotomised (teno group), tenotomised and subjected to manual acupuncture at ST36 (ST36 group), BL57 (BL57 group) and ST36+BL57 (SB group) and EA at ST36+BL57 (EA group). The mass-average diameter (MAD) and the reorganisation of collagen fibril diameters were determined during the three phases of tendon healing (at 7, 14 and 21 days). RESULTS: The MAD increased during the three phases of healing in the SB group. In the EA group, MAD increased initially but was reduced at day 21. The reorganisation of collagen fibrils was improved in the EA and SB groups at days 14 and 21, respectively. EA at day 21 appeared to reduce the reorganisation. CONCLUSIONS: These results indicate that the use of EA up to day 14 and manual acupuncture at ST36+BL57 up to day 21 improve the ultrastructure of collagen fibrils, indicating strengthening of the tendon structure. These data suggest a potential role for acupuncture in rehabilitation protocols.

Green Brazilian propolis effects on sperm count and epididymis morphology and oxidative stress.

Basal generation of reactive oxygen species (ROS) is essential for male reproductive function, whereas high ROS levels may be linked to low quality of sperm and male infertility. The number of antioxidants known to inflict damage is growing, and it will be of interest to study natural products, which may have this activity. Since the epididymis is known to play an important role in providing the microenvironment for sperm maturation and storage of sperm, this study was undertaken to evaluate the morphometric-stereological and functional alterations in the epididymis after chronic treatment with low doses of Brazilian green propolis, which is known for its antioxidant properties. For this purpose, forty-eight adult male Wistar rats were treated with 3, 6 and 10 mg/kg/day of aqueous extract of Brazilian green propolis during 56 days and morphological parameters, sperm production and number of sperm in rat epididymis and oxidative stress levels were analyzed. The results showed higher sperm production and greater epithelium height of the epididymis initial segment and no induction of oxidative stress in treated animals. Further studies are needed to fully understand the effects of propolis on the reproductive system but our results showed that it could alter male reproductive function.

Light microscope observation of circulating human lymphocytes cultured in vitro

The purpose of this work was to study the isolation and a light microscopy technique for cultured lymphocytes. Blood samples were obtained by venipuncture with an anticoagulant added and centrifuged in a Percoll density gradient to separate the leukocytes. Lymphocytes were placed in 25 cm ³ tissue culture flasks at 37ºC. After culturing, they were fixed and stained with the methods used for blood smears. Results showed that not all fixing solutions and stains were an equally good choice for cultured lymphocytes.

Os linfócitos são células importantes do sistema imune e têm sido largamente utilizados em estudos morfológicos. Entretanto, a literatura sobre técnicas de preparação dessas células é escassa e antiga, especialmente para linfócitos cultivados in vitro. Portanto, o objetivo desse estudo foi relatar com detalhes as técnicas de isolamento e microscopia de luz de linfócitos mantidos em cultura. Amostras de sangue foram obtidas por punção venosa e centrifugadas em gradiente de densidade de Percoll, para separar os leucócitos. Os linfócitos foram mantidos em frascos de cultura de 25 cm³ a 37ºC. Após a cultura, as células foram fixadas e coradas de acordo com a metodologia utilizada para esfregaços sanguíneos. Nossos resultados mostraram que nem todos os fixadores e corantes utilizados para esfregaços sanguíneos são uma boa escolha para linfócitos cultivados in vitro.