Detailed Morphological Analysis of Cryoinjury in Human Ovarian Tissue Following Vitrification or Slow Freezing.

Cryopreservation of human ovarian tissue represents a key procedure for fertility preservation. The two most widely used cryopreservation methods for human ovarian cortex samples are slow freezing\thawing (SF\T) and vitrification\warming (V\W). The aim of the present study was to analyze the effects of SF\T and V\W using a metal chamber, on specific follicle and oocyte structures and on the stromal organization post-cryopreservation. We did histology analysis of SF\T and V\W ovarian fragments from nine healthy subjects. Overall results showed that cryopreserved tissues presented significant rates of damage in primordial and primary follicles. Altered nuclear structure of primordial follicles and cell detachment from primordial and primary follicles were the main injuries observed after V/W and SF/T. The stromal components were similarly well preserved after cryopreservation. We conclude that both cryopreservation methods may be used for fertility preservation purposes with similar outcomes in terms of follicular and stromal integrity. Detachment of follicle cells from basal membrane represents an important cryoinjury that deserves further investigation.

The use of parthenotegenetic and IVF bovine blastocysts as a model for the creation of human embryonic stem cells under defined conditions.

PURPOSES: Clinical application of human embryonic stem cells will be possible, when cell lines are created under xeno-free and defined conditions. We aimed to establish methodologies for parthenogenetic activation, culture to blastocyst and mechanical isolation of the inner cell mass (ICM) using bovine oocytes, as a model for derivation and proliferation of human embryonic stem cells under defined xeno-free culture conditions. METHODS: Cumulus-oocyte-complexes were in vitro matured and activated using Ca(2+)Ionophore and 6-DMAP or in vitro fertilized (IVF). Parthenotes and biparental embryos were cultured to blastocysts, when their ICM was mechanically isolated and placed onto a substrate of fibronectin in StemPro medium. After attachment, primary colonies were left to proliferate and stained for pluripotency markers, alkaline phosphatase and Oct-4. RESULTS: Parthenogenesis and fertilization presented significantly different success rates (91 and 79 %, respectively) and blastocyst formation (40 and 43 %, respectively). ICMs from parthenogenetic and IVF embryos formed primary and expanded colonies at similar rates (39 % and 33 %, respectively). Six out of eight parthenogenetic colonies tested positive for alkaline phosphatase. Three colonies were analyzed for Oct-4 and they all tested positive for this pluripotency marker. CONCLUSION: Our data show that Ca(2+) Ionophore, and 6-DMAP are efficient in creating large numbers of blastocysts to be employed as a model for human oocyte activation and embryo development. After mechanical isolation, parthenogetic derived ICMs showed a good rate of derivation in fibronectin and Stem-Pro forming primary and expanded colonies of putative embryonic stem cells. This methodology may be a good strategy for parthenogenetic activation of discarded human oocytes and derivation in defined conditions for future therapeutic interventions.

Investigation of the faecal microbiota of geriatric cats.

AIMS: Aim of the study was to investigate the faecal microbiota of geriatric cats, as aging affects the nutrient digestibility and metabolic function of the feline intestine. METHODS AND RESULTS: Twenty geriatric cats were randomly assigned to two groups that were fed different foods. Coriobacteriaceae, Clostridium cluster XIV, bifidobacteria and lactic acid bacteria were the dominant faecal bacterial groups, accounting for c. 40% of total bacteria. Clostridium cluster IX was less predominant (0.5% of total bacteria), while the remaining bacterial populations enumerated only accounted for 0.2% of total bacteria. Highly diverse microbial profiles were demonstrated for geriatric cats with denaturing gradient gel electrophoresis, although a few common bands were evident. Some differences were seen in the feline faecal microbiota between animal groups at the same time or over time for individual animals. However, no obvious clustering based on animal group or sample time was indicated. CONCLUSIONS: Geriatric cats harboured a complex faecal microbiota and c. 41% of total bacteria have been detected with the probes employed. SIGNIFICANCE AND IMPACT OF THE STUDY: First molecular-based study examining faecal microbiota of geriatric felines. Knowledge of the microbiota associated with ageing in cats may allow improved development of foods specific for the needs of senior cats.

The effects of sample size on the outcome of ovarian tissue cryopreservation.

Cryopreservation of ovarian tissue is known to affect follicular survival. Several variables may be responsible for this. Little attention has focused on the effect of the size of the fragment to be cryopreserved. This study was conducted to assess the effect of the size of the tissue on follicular histology after freezing with 1,2-propanediol. Histological evaluations were performed of control and cryopreserved tissue. Fragments were cut 10 x 3 x 2 mm(3) (2 mm group) or 10 x 3 x 4 mm(3) (4 mm group). Percentages of normal follicles in control fragments cut into 2 and 4 mm slices were 56% and 34%, respectively. The relative risks to obtain normal follicles in the 2 mm and the 4 mm fragments after cryopreservation were 0.63 and 0.47, respectively. Freezing reduced follicle survival to a significantly greater extent in the larger tissue fragments. There is an increased risk of damage to primary and primordial follicles, when the tissue slices are cut with all dimensions larger than 2 mm.

Color Doppler sonographic study of the uterine artery in patients using intrauterine contraceptive devices.

A transvaginal Doppler sonographic examination of uterine artery blood flow was performed. Resistance and pulsatility indices were measured in 101 women, 74 of whom were using an intrauterine contraceptive device, and 27 controls, who were not using any contraceptive method. The intrauterine contraceptive device users were divided into three groups: those with normal bleeding (n = 34); those with abnormal uterine bleeding without medication (n = 16); and those with abnormal bleeding corrected with use of prostaglandin inhibitors (n = 24). The resistance and pulsatility values were significantly lower in the group of women using intrauterine contraceptive devices who had abnormal bleeding than in all other groups (P<0.001). All other comparisons were nonsignificant. A pulsatility index of less than 2 may be associated with a higher risk for development of intrauterine contraceptive device-induced bleeding.

Diagnóstico pré-natal por punçäo da vilosidade coriônica / Prenatal diagnosis by chorionic villus sampling

Resultado de 129 punçöes da vilosidade coriônica para diagnóstico pré-natal no primeiro trimestre de gestaçäo. Os métodos de rotina par preparaçäo direta, preparaçäo de 24 horas e cultura de longa duraçäo, bem como as indicaçöes säo descritos detalhadamente. Tendo em vista a precocidade desse exame e alto sucesso de técnica (97,4%), pode-se concluir pela sua aceitabilidade como método complementar de diagnóstico genético pré-natal

Differences in behavioral responses to oxotremorine and physostigmine in New Zealand black (NZB/BlNJ) and C57BL/6 mice.

The NZB/BlNJ (NZB) mice are an autoimmune-prone strain, known to develop brain-reactive antibodies in serum at much earlier chronological ages than normal mice. Measurement of locomotor activity in 8-10 month old C57BL/6 (C57) mice following the administration of either oxotremorine or physostigmine, revealed a biphasic response consisting of inhibition at small doses, but increased motor activity at large doses. In contrast, age-matched NZB mice exhibited little inhibition at the smaller doses, but had much greater increases in activity after the larger doses. Similarly, when compared to C57 mice, NZB mice were less sensitive to oxotremorine-induced salivation, diarrhea and visible tremors. Moreover, oxotremorine-induced hypothermia occurred at smaller doses in C57 mice than in NZB mice and was of a greater magnitude. Thus, at an age when NZB mice possess high levels of brain-reactive antibodies, and exhibit impairment in tests of learning/memory, these mice also show diminished responses in several tests of cholinomimetic-induced behavior and physiological alterations.

[Sciatica caused by synovial cysts and lumbar articular diverticula with intraspinal development. Saccoradiculographic, computed tomographic and arthrographic study]. / La sciatique par kystes synoviaux et diverticules articulaires lombaires à développement intra-rachidien. Etude saccoradiculographique, tomodensitométrique et arthrographique.

The authors report 8 cases of patients with synovial cysts (3 cases) or diverticuli (5 cases) of the lumbar joints, compressing the sciatic roots. They describe the radiological aspects of the two sides of the degenerative pathology of the posterior articular processes, which are relatively unrecognized (14 cases in the literature). They insist on the advantage of saccoradiculography which is the only test to be able to demonstrate dynamic alterations in relation with diverticuli, if performed in a standing position. The authors show the advantage of tomodensitometry which supports the diagnosis of cyst in the presence of a fibrous wall image. They emphasize the value of the arthrography of the posterior joint; it presents a diagnostic role by visualizing the cavities, and a therapeutic role by permitting the injection in situ of steroids and/or local anesthetics.

[Posterior lumbar vertebral arthrography. Pathologic aspects]. / Arthrographie articulaire vertébrale postérieure lombaire. Aspects pathologiques.

Findings on arthrography of posterior lumbar spine joints in 55 patients were compared with those reported in the literature. Two major degenerative lesions of these joints, diverticula and cysts, can provoke back pain and symptomatic nerve compression. Whereas diagnosis of a cyst is dependent mainly on computed tomography imaging, that of a diverticulum is ensured by arthrography, an investigation which also has a therapeutic aim since it allows infiltrations to be performed.

[Posterior lumbar vertebral arthrography. Normal anatomic study]. / Arthrographie vertébrale lombaire postérieure. Etude anatomique normale.

Historical aspects and anatomophysiologic features of posterior lumbar vertebral joints are summarized and results of arthrography of 55 joints presented, together with a simple, original method for performing this examination. Normal anatomic appearances condition understanding of principal lesions encountered, particularly with respect to compression of adjacent nerve structures.

Corynebacterium JK: a cause of nosocomial device-related infection.

Ten cases of serious infection due to antibiotic-resistant Corynebacterium JK are reported. Five infections occurred in patients with hematologic malignancy and granulocytopenia, an association previously reported. Five additional nosocomial infections occurred in patients without underlying disease previously reported to be associated with JK. Nine of the 10 infections were nosocomial in origin and were associated with plastic devices that had been inserted into the patients. JK organisms were isolated from a variety of sources from an additional 40 uninfected patients. Although the majority of clinical isolates of Corynebacterium JK were contaminants, our experience suggests that as many as 20% of isolations are associated with serious nosocomial infections. Isolation of Corynebacterium JK--particularly in blood cultures--from patients with indwelling catheters or other devices is highly suggestive of the presence of a serious nosocomial infection.