The physical activity experience of prostate cancer patients: a multicentre peer motivation monitoring feasibility study. The Acti-Pair study.

BACKGROUND: Although the benefits of physical activity (PA) on health are recognised, prostate cancer patients do not follow PA recommendations. The barriers to PA, whether physical, environmental or organisational, are known. Furthermore, even when such barriers are overcome, this achievement is not systematically accompanied by a change in lifestyle habits. The proposal of a programme enabling the integration of PA in the patient's everyday life represents a new challenge in the personalized management of cancer patients. Peer-mentoring interventions have demonstrated their effectiveness in increasing adherence to PA by patients. This study aimed (1) to assess the feasibility of a peer-mentoring intervention: the Acti-Pair program in a local context and (2) to assess the effectiveness of the intervention in this context. METHODS AND ANALYSIS: A pre-post  design pilot study will be used to evaluate feasibility, potential effectiveness and implementation outcomes overs in prostate cancer patients. We performed a mixed quantitative and qualitative prospective study to assess means and process indicators and the implementation of the Acti-Pair program. This study will be performed in cancer centres of Loire district and will be comprised of three successive stages (1) diagnosis of the target population, (2) recruitment and training of peers, and (3) implementation of this intervention in the Loire department. DISCUSSION: This study will allow us to extend the peer-mentoring intervention to other contexts and assess the effectiveness of this intervention and its generalisability.

[Epidemiology of patients with eye-related emergencies in a university hospital: A cross-sectional study identifying factors associated with true emergency]. / Caractéristiques des patients examinés en urgence par un ophtalmologiste dans un CHU : résultats d'une étude transversale analytique identifiant des facteurs associés au caractère de véritable urgence.

PURPOSE: To describe epidemiological characteristics of outpatients examined by university medical center ophthalmologists in emergency rooms (ER), and to determine factors associated with true emergencies. METHODS: A monocentric cross-sectional study including all patients examined by an ophthalmologist in the ER of in the university hospital of Nancy over a two-month period was conducted. Demographics and medical characteristics were assessed. The visits were categorized by ophthalmologists as true emergencies or not. RESULTS: Among the 1,308 patients included, the median (IQR) age was 49 (32-64) years, and 56 % were males. The main reasons for seeking care were eye redness (32.6 %), eye pain (30.0 %), eye trauma (26.1 %), and visual loss (23.3 %). Nearly 40 % of the consultations were judged as not truly emergent. Factors significantly associated with true emergencies were: age under 60, male gender, some reasons for seeking care (visual loss, eye redness, eye pain), and a period of less than 3 days between symptom occurrence and the ER visit. CONCLUSION: The proportion of non-emergent ER visits was relatively high, and factors associated with true emergencies have been identified in our study. Standardized protocols may be useful to help emergency physicians and nurses to determine when to refer a patient to an ophthalmologist.

Application of CRISPR/Cas9 to Autophagy Research.

The ability to efficiently modulate autophagy activity is paramount in the study of the field. Conventional broad-range autophagy inhibitors and genetic manipulation using RNA interference (RNAi), although widely used in autophagy research, are often limited in specificity or efficacy. In this chapter, we address the problems of conventional autophagy-modulating tools by exploring the use of three different CRISPR/Cas9 systems to abrogate autophagy in numerous human and mouse cell lines. The first system generates cell lines constitutively deleted of ATG5 or ATG7 whereas the second and third systems express a Tet-On inducible-Cas9 that enables regulated deletion of ATG5 or ATG7. We observed the efficiency of autophagy inhibition using the CRISPR/Cas9 strategy to surpass that of RNAi, and successfully generated cells with complete and sustained autophagy disruption through the CRISPR/Cas9 technology.

Improved cryopreservation of in vitro-produced bovine embryos using a chemically defined freezing medium.

This study evaluates a new synthetic substitute (CRYO3, Ref. 5617, Stem Alpha, France) for animal-based products in bovine embryo cryopreservation solutions. During the experiment, fetal calf serum (FCS) and bovine serum albumin (BSA) were used as references. A combination of a thermodynamic approach using differential scanning calorimetry and a biological approach using in vitro-produced bovine embryo slow-freezing was used to characterize cryopreservation solutions containing CRYO3, FCS and BSA. The CRYO3 and fetal calf serum (FCS) slow-freezing solutions were made from Dulbecco's phosphate-buffered saline containing 1.5 m ethylene glycol, 0.1 m sucrose and 20% (v.v(-1)) of CRYO3 or FCS. The bovine serum albumin (BSA) solution was made by adding 0.1 m sucrose to a commercial solution containing 1.5 m ethylene glycol and 4 g L(-1) BSA. These solutions were evaluated using three characteristics: the end of melting temperature, the enthalpy of crystallization (thermodynamic approach) and the embryo survival and hatching rates after in vitro culture (biological approach). The CRYO3 and FCS solutions had similar thermodynamic properties. In contrast, the thermodynamic characteristics of the BSA solution were different from those of the FCS and CRYO3 solutions. Nevertheless, the embryo survival and hatching rates obtained with the BSA and FCS solutions were not different. Similar biological properties can thus be obtained with slow freezing solutions that have different physical properties within a defined range. The embryo survival rate after 48 h of in vitro culture obtained with the CRYO3 solution (81.5%) was higher than that obtained with the BSA (42.2%, P = 0.000 12) and FCS solutions (58%, P = 0.016). Similarly, the embryo hatching rate after 72 h of in vitro culture was higher with the CRYO3 solution (61.1%) than with the BSA (31.1%, P = 0.0055) and FCS solutions (36%, P = 0.018). We conclude that CRYO3 can be used as a chemically defined substitute for animal-based products in in vitro-produced bovine embryo cryopreservation solutions.

Conjugated quantum dots allow for the co-localization of endogenous proteins in submicroscopic structures of the rat cerebellum.

GABA and glutamate are known as the principal inhibitory and excitatory neurotransmitters in the vertebrate central nervous system, respectively. However, recent electro-physiological and immunogold literature data indicate that GABA may undergo also an excitatory action on presynaptic varicosities of parallel fibers (PFs) in the molecular layer of the rat cerebellum. PFs are axonal extensions, with a cross section of about 0.1 microm, of the glutamatergic granule cells. Such an unexpected excitatory action of GABA indicates clearly the presence of GABA receptors in the PFs of granule cells. We show in this study that quantum dots may be used specifically and efficiently to label two endogenous synaptic proteins, namely R-GABA(A)-alpha1 receptors (GABA(A) Rs) and glutamate transporters (VGLUT1) in order to target their localization in very small structures such as the presynaptic varicosities of the PFs.

[Vitrification: a future technique for ovarian cryopreservation? Physical basis of cryobiology, advantages and limits]. / La vitrification : technique d'avenir pour la cryoconservation ovarienne ? Bases physiques de cryobiologie, avantages et limites.

Ovarian cryopreservation is presently indicated in patients who undergo a gonadotoxic treatment, most commonly for anticancer procedures. These procedures can strongly alter fertility by damaging the follicular ovarian reserve. Although six human live births have been described in the world after ovarian tissue cryopreservation and autografting, the techniques of cryopreservation techniques are not consensual. Vitrification is a physical process that allows cryopreservation without formation of ice crystals, by transformation of a highly concentrated solution in a glassy or amorphous state. Vitrification is at present rapidly expanding in the biology of reproduction. With the classic methods of freezing, formation of ice crystals within the ovarian tissue is systematic and can entail cellular lesions. Which is why more and more teams question the theoretical advantage of the vitrification for ovarian cryopreservation. Our objective was to summarize the fundamental physical basis of cryobiology, necessary for an understanding of vitrification. From our experience, we also wanted to point out the practical difficulties of this technique, and we are proposing a model of evaluation and validation that uses differential scanning calorimetry, applicable to any protocol of vitrification.

The tyrosine kinase Syk regulates the survival of chronic lymphocytic leukemia B cells through PKCdelta and proteasome-dependent regulation of Mcl-1 expression.

B-cell chronic lymphocytic leukemia (B-CLL) is characterized by accumulation of mature monoclonal CD5+ B cells. The disease results mainly from a failure of cells to undergo apoptosis, a process largely influenced by the existence of constitutively activated components of B-cell receptor signaling and the deregulated expression of anti-apoptotic molecules. Recent evidence pointing to a critical role of spleen tyrosine kinase (Syk) in ligand-independent BCR signaling prompted us to examine its role in primary B-CLL cell survival. We demonstrate that pharmacological inhibition of constitutive Syk activity and silencing by siRNA led to a dramatic decrease of cell viability in CLL samples (n=44), regardless of clinical and biological status and induced typical apoptotic cell death with mitochondrial failure followed by caspase 3-dependent cell death. We also provide functional and biochemical evidence that Syk regulated B-CLL cell survival through a novel pathway involving PKCdelta and a proteasome-dependent regulation of the anti-apoptotic protein Mcl-1. Together, our observations are consistent with a model wherein PKCdelta downstream of Syk stabilizes Mcl-1 through inhibitory phosphorylation of GSK3 by Akt. We conclude that Syk constitutes a key regulator of B-CLL cell survival, emphasizing the clinical utility of Syk inhibition in hematopoietic malignancies.

Towards whole sheep ovary cryopreservation.

Cryopreservation of ovarian tissue aims to assist young women who require treatments that may lead to sterility or infertility. Cryopreservation procedures should therefore be as simple and efficient as possible. This study investigates rapid cooling outcomes for whole sheep ovaries. Ovaries were perfused with VS4 via the ovarian artery, and cooled by quenching in liquid nitrogen in less than a minute (estimated cooling rate above 300 degrees C/min till the vitreous transition temperature). The ovaries were rewarmed in two stages: slow warming (12-16 degrees C/min from -196 to -133 degrees C) in liquid nitrogen vapour, followed by rapid thawing in a 45 degrees C water bath at about 200 degrees C/min. DSC measurements showed that under these cryopreservation conditions VS4 would vitrify, but that VS4 perfused ovarian cortex fragments did not vitrify, but formed ice (around 18.4%). Immediately following rewarming, a dye exclusion test indicated that 61.4+/-2.2% of small follicles were viable while histological analysis showed that 48+/-3.8% of the primordial follicles were normal. It remains to be clarified whether follicle survival rates will increase if conditions allowing complete tissue vitrification were used.

[Ovarian tissue vitrification: Cortex and whole ovary in sheep]. / Vitrification du tissu ovarien: cortex et ovaire entier chez la brebis.

OBJECTIVE: The aim of this study was to evaluate a cryopreservation technique by vitrification of cortex or whole ovaries in sheep, using two cryoprotectant solutions: VS1 and VS4 and to study their physical properties to avoid ice crystallisation by vitrification of whole sheep ovaries permeated with a cryoprotectant solution. ANIMALS AND METHODS: From 6-month-old ewes, whole sheep ovaries with their vascular pedicles were collected at the slaughterhouse or at the veterinary school and prepared for cryoprotectant toxicity tests and freezing procedure. Follicle viability was measured by trypan blue test and histological examination of ovary. The hemi-ovarian cortex was stored in liquid nitrogen. Four to six weeks after the first laparotomy, the controlateral ovary was removed and the vitrified-warmed hemi-ovary was sutured. Thermal properties of a cryoprotectant solution called VS4 (critical cooling rates [Vccr], vitreous transition temperature [Tg], end of melting temperature [Tm]) were measured by differential scanning calorimetry. RESULTS: No significant difference in follicle viability or normal follicle rates was observed between ovarian cortex exposed or non-exposed to cryoprotectant solutions. Nor was any significant difference observed before and after vitrification. Three pregnancies occurred, from which four lambs were born after autografts of vitrified ovarian cortex. With whole ovary, the decrease in the number of normal follicles was lower when frozen-thawed ovaries were treated with VS4 (P = 0.04). There were less nuclear anomalies (P = 0.02). The Vccr of VS4 has been estimated to be 14.3+/-1.1 degrees C/min and Tg was -125.0+/-0.2 degrees C. Because the penetration of cryoprotectants was very low, Vccr was very high and the cooling speed did not allow cortex to vitrify. DISCUSSION AND CONCLUSIONS: Cryopreservation of cortex or whole ovary by vitrification seems a promising technique in reproductive medicine. The best histologic results were obtained with the VS4 cryoprotectant when whole ovary was vitrified.

Thermal properties of ethylene glycol aqueous solutions.

Preventing ice crystallization by transforming liquids into an amorphous state, vitrification can be considered as the most suitable technique allowing complex tissues, and organs cryopreservation. This process requires the use of rapid cooling rates in the presence of cryoprotective solutions highly concentrated in antifreeze compounds, such as polyalcohols. Many of them have already been intensively studied. Their glass forming tendency and the stability of their amorphous state would make vitrification a reality if their biological toxicity did not reduce their usable concentrations often below the concentrations necessary to vitrify organs under achievable thermal conditions. Fortunately, it has been shown that mixtures of cryoprotectants tend to reduce the global toxicity of cryoprotective solutions and various efficient combinations have been proposed containing ethanediol. This work reports on the thermal properties of aqueous solutions with 40, 43, 45, 48, and 50% (w/w) of this compound measured by differential scanning calorimetry. The glass forming tendency and the stability of the amorphous state are evaluated as a function of concentration. They are given by the critical cooling rates v(ccr)above which ice crystallization is avoided, and the critical warming rates v(cwr) necessary to prevent ice crystallization in the supercooled liquid state during rewarming. Those critical rates are calculated using the same semi-empirical model as previously. This work shows a strong decrease of averaged critical cooling and warming rates when ethanediol concentration increases, V(ccr) and V(cwr) = 1.08 x 10 (10) K/min for 40% (w/w) whereas V(ccr) = 11 and V(cwr) = 853 K/min for 50% (w/w). Those results are compared with the corresponding properties of other dialcohols obtained by the same method. Ethylene glycol efficiency is between those of 1,2-propanediol and 1,3-propanediol.

A simple capacitive cell for the measurement of liquids dielectric constant under transient thermal conditions.

A simple device for the measurement of the complex dielectric permittivity of liquids in various thermodynamic states has been developed. It uses a cylindrical aluminium capacitor of a type currently applied in tuning antenna circuits. The capacitor is filled with the liquid solution under study. A comparison of its capacity is made with that of the nitrogen filled capacitor tested under the same thermal conditions. This comparison allows the determination of the real and imaginary part of the solutions permittivity as a function of temperature (between 150 and 300 K) and frequency (between 100 Hz to 2 MHz). After validating the technique with pure glycerol and pure 1,2-propanediol, spectroscopic measurements have been undertaken on pure and diluted 1,2-propanediol in water. Due to the low heat capacity and the high thermal conductivity of the capacitor, cooling rates of 40 K/min have been achieved inside the solution, allowing measurements in the supercooled liquid and vitreous states. Results are presented and discussed in terms of relaxation and the physical states of the sample. By selecting the required thermal conditions, this device permits the observation of thermal transitions, such as ice crystallisation, and measurements to be conducted in the unstable supercooled liquid state. These measurements are necessary in the development of an effective electromagnetic warming device for vitrified cryoprotective solutions.

Glass-forming tendency in the system water-dimethyl sulfoxide.

The glass-forming tendency on cooling and the stability of the wholly amorphous state on warming have been previously reported for many cryoprotective solutions. However, unlike the other solutions, those of dimethyl sulfoxide (Me(2)SO) have not been studied on cooling. In this paper, the glass-forming tendency of Me(2)SO aqueous solutions has been measured for solutions containing 40, 43, 45, and 47.5% (w/w) Me(2)SO. At a concentration of 45% (w/w), the glass-forming tendency decreases in the following order: levo-2, 3-butanediol, 1,3-butanediol, 1,2-propanediol, 1,2,3-butanetriol, dimethyl sulfoxide, dimethylformamide, diethylformamide, 1, 4-butanediol, ethylene glycol, glycerol, 1,3-propanediol. New measurements have also been made on warming the Me(2)SO solutions.

Glass-forming tendency and stability of aqueous solutions of diethylformamide and dimethylformamide

The glass-forming tendency on cooling and the stability of the wholly amorphous state on warming of aqueous solutions of diethylformamide and of dimethylformamide have been studied by calorimetry. With diethylformamide, only ice formation is observed except on warming at the lowest rate of 2.5 degreesC/min, where occasionally a hydrate forms also. The hydrate was observed up to 10 degreesC/min with 50% diethylformamide. With dimethylformamide hydrates form even at high warming rates. The last hydrate melts at -47.7 degreesC. The warming thermograms are much more complicated than for diethylformamide. For the glass-forming tendency on cooling, as well as for the stability of the wholly amorphous state on warming, these two compounds, at concentrations of 40, 45, or 50% (w/w) in water, are more efficient than glycerol and ethylene glycol, but less than 1,2-propanediol and levo-2,3-butanediol. On warming, they are comparable to DMSO. Pure diethylformamide could not be crystallized, whereas, conversely, pure dimethylformamide could not be vitrified. Curiously, the glass transition of aqueous solutions of diethylformamide increases and then decreases with the diethylformamide concentration in water, contrary to other cryoprotectants, for which it always increases or decreases. Diethyl- and dimethylformamide could be interesting cryoprotectants if they are not too toxic when added before cryopreservation, and in the case of dimethylformamide, if one can avoid damage due to its hydrates. Copyright 1998 Academic Press.

Critical cooling and warming rates to avoid ice crystallization in small pieces of mammalian organs permeated with cryoprotective agents.

Measurements were made by differential scanning calorimetry on small pieces of rabbit kidney permeated with 2, 3-butanediol containing mainly the levo- and dextro-isomers. The critical cooling rates necessary to vitrify the pieces of organ, and the corresponding critical warming rates which are required to avoid crystallization in the vitrified samples, were determined. The dynamic method used for these determinations is described. The glass-forming tendency and the stability of the amorphous state were both greater in the kidney tissue samples than in the bulk cryoprotective solution. This result is discussed in the context of the lowering of the freezing point of water in emulsions and the promotion of supercooling in hydrogels and porous materials. In corresponding experiments with rat hearts impregnated with 1,2-propanediol, only the critical warming rate was reduced.

Effect of Saccharides on the Glass-Forming Tendency and Stability of Solutions of 2,3-Butanediol, 1,2-Propanediol, or 1,3-Butanediol in Water, Phosphate-Buffered Saline, Euro-Collins Solution, or Saint Thomas Cardioplegic Solution

The effect of sugars or reduced saccharides trehalose, sucrose, sorbitol, or mannitol on the glass-forming tendency during cooling and the stability of the wholly amorphous state during warming has been studied with 2,3-butanediol, 1,2-propanediol, or 1,3-butanediol in three different carrier solutions. The 2,3-butanediol contained 96.7% (w/w) racemic mixture of the levo and dextro isomers and 3.1% (w/w) of the meso isomer (called 2,3-butanediol 97% dl). The carrier solutions were water, a phosphate-buffered saline, and two organ preservation solutions (Euro-Collins and Saint Thomas). The latter two were chosen because they are often used for kidney and heart preservation, respectively. The concentrations of 2,3-butanediol, 1,2-propanediol, and 1,3-butanediol varied respectively from 25 to 34, 30 to 35, and 30% (w/w). The concentrations of saccharides were 4 or 5% (w/w). In the absence of saccharides, for a given 2,3-butanediol concentration, the glass-forming tendency increased in the following order: water, Saint Thomas, the phosphate buffer, Euro-Collins. Addition of 4 or 5% (w/w) saccharide resulted in a large increase in the glass-forming ability of the solution during cooling and increased the stability of the glass during warming; but replacement of 4 or 5% diol by an equivalent weight (percentage) of a saccharide decreased, though to a lesser extent, these properties.

[Organ cryopreservation: current status of research carried out in Grenoble]. / Cryopréservation d'organes: etat actuel des recherches développées à Grenoble.

After discussing the problem of organ cryopreservation and reviewing the current data available on this subject, the Grenoble project is presented. Physical and biological studies have been combined with experimentation of autologous renal transplantation in rabbits to assess the functional value of the retransplanted organ after treatment and cooling. Renal resistances are measured during perfusion of the kidney with the cryoprotective solution. In order to verify the homogeneity of the cryoprotector concentration in the organ, on NMR spectral imaging test has been developed. A new rapid imaging method now allows real time monitoring of concentration variations during perfusion. In addition to concentration and homogeneity, analysis of local spectra also provides information about the local temperature de the kidney.